STUDIES ON INDIAN HORSESHOE CRABS

(Merostomata:Xiphosura) WITH SPECIAL

REFERENCE TO ITS FEEDING BEHAVIOUR

THESIS SUBMITTED FOR THE DEGREE OF

DOCT
DOCTOROR OF PHIL OSOPHY (SCIENCE)
PHILOSOPHY
OF THE
UNIVERSITY OF CAL CUTT
CALCUTT
CUTTAA

RAKHAL DEBN
DEBNAATH
M.Sc. (Zool.), NET, M.Phil. (Env. Sci.)

DEPAR
DEPAR TMENT OF MARINE SCIENCE
ARTMENT
UNIVERSITY OF CALCUTT
CALCUTT A
CUTTA
35 , B ALL
BALL YGUNGE CIR
ALLY CULAR R
CIRCULAR ROOAD
CAL CUTT
CALCUTT
CUTTA A - 700019,WEST BENGAL
I N D I A
JUNE , 1992
 Dedicated to My
Dedicated

late
late

Mother
&
Father

*** First Ph.D. work on Indian Horseshoe Crabs(June'1992)

All rights reserved. No part of this Thesis

© Dr. R. Debnath,
may be reproduced, stored or transmitted,
6th April' 2005
in any form or by any means, without the
Electronic Publication
permission of the author.
 C O N T E N T S
PAGE NO.
1. Acknowledgement.......................................4 - 5
2. Preamble...................................................... 6

3. Historical Resume.......................................7 - 8
4. Physiography & Hydrology........................9 - 14
5. Methodology................................................15 - 19
6. Part-1. Studies on Population Aspects
A. Geographic Distribution & Habitat..................................................20 - 25
B. Morphology & Morphometry...........................................................26 - 53
C. Absolute Population Estimates.........................................................54 - 83
D. Breeding Activity, Sex-ratio & Predation.......................................84 - 104
E. Epifaunal Associates......................................................................105 -114

7. Part-II. Studies on Food,Feeding & Digestion

A. Digestive System............................................................................115 - 119
B. Gut Content Analysis.....................................................................119 - 122
C. Feeding, Digestive Physiology & Trophic Level...........................122 - 133

8. Part-III. Studies on Central Nervous System

A. Anatomy of The Central Nervous System.....................................134 - 136

B. Cytomorphology of CNS ..............................................................136 - 142

(with Special reference to its
Neurosecretory Elements)

9. Summary ...................................................143
10. References .................................................144 - 158
11. Published Papers (not included in this electronic edition)
 4

AC K N OWLE D GE M E NT
I express my deepest sense of gratitude to my revered teacher , Prof. Dr. Amalesh
Choudhury , Department of Marine Science , University of Calcutta ,for his learned
counsel ,affectionate guidance , sustained interest and constant encouragement during the
course of this investigation as also in preparation of the manuscript .

My sincere acknowledgements are due to the authorities of the Susama Devichoudhurani

Marine Biological Research Institute, Sagar Island , West Bengal , India for the
invaluable facilities which I enjoyed during the investigation of my research project .

I am fortunate to acknowledge Dr. B.K.Tikadar , the then Director , Zoological Survey

of India , Calcutta , for his generous suggestions making an impetus to start the present
investigation in the year 1984 .

I am thankful to the Head of the Department of Marine Science , University of

Calcutta for providing all sorts of laboratory and library facilities during the tenure of
this work. I am also thankful to the Research Scholars of this department for their
cheerful help and interest in course of this dissertation work .

I would also like to extend my sincere thanks to Dr. P.S.Choudhury and Dr. D.K.
Nanda of the Department of Zoology , University of Calcutta for their help during the
study on neuroendocrine system and preparation of the manuscript incoroporated in Part-III
of the present thesis .

I would like to acknowledge my indebtness to Prof. Dr. Carl N. Shuster , Jr. ,

Virginia Institute of Marine Science , USA , for kindly offering me the benefit of his long
experience with Limulus and pains taking review of my manuscript delt with breeding
behaviour of Indian horseshoe crabs . I am also greateful to Dr. M.L. Botton, Fordham
University , USA , and Dr. H . Sugita , University of Tsukuba , Japan , for providing me
reprints , books and valuable suggestions during the entire course of the present
investigation , without which I could not present this thesis .

I greatefully acknowledge the Department of Forest , Govt. of Orissa and

the authorities of Paradwip Port Trust for their help and co-operation in course of the
study .

I am thankful to Sri Jalad Gayen , Sri Jiten Singh , Sri Sadhan Sutradhar and
Sri Sisir Kr. Roy for assisting me during coastal tours and field investigations . I also
acknowledge Sri Siben DebSinha for his assistance in arranging the reference column .

I must acknowledge the University Grants Commission and the Department of

Environment , New Delhi , for financial assistance .

This is also an occasion to acknowledge the Directorate of Higher Education ,

Govt. of Tripura , India for sanctioning my station leave permission during the
preparation of this thesis paper .

It is a great pleasure to me to express my heartiest thanks to Dr.R.K. Sinha , and

Mr. S. Kundu,
Kundu University of Calcutta , who helped tremendously to give a final shape of
this thesis .

Lastly I wish to record a deep sense of gratitude to my mother and also to my beloved
wife for their good wishes and encouragement which always inspire me to go ahead .

sd/- Illegible
Dated, the 8 th June,1992 _____________________
CAL CUTT
CALCUTT
CUTTAA , INDIA ( RAKHAL DEBNATH )
Tachypleus gigas , with a bizzare appearance and
horseshoe shaped body is one of the four extant species of the
present world , basically unchanged for last 200 million
years . The animal had noticed the drifting of Continental
Shelf and the rising mountains . It roams along the coast line
of Bay of Bengal sharing the space with other closest
relative , Tachypleus ( = Carcinoscorpius , n. comb.)
rotundicauda. For mankind , study of the hemolymph and
eye of these has led to medical breakthrough . Care should
be taken immediately to protect these unique and harmless
marine creatures , considered now as the " threatened " and
"endangered " species from the verge of extinction .
 6

PREAMBLE

The horseshoe crabs or "King crabs" ( an erroneous term) are regarded as one of the most primitive
animals existing in today's world , dating back to the period of 200 millons years . They are basically
marine animals occurring on sandy and muddy bottoms from the shore lines to a depth of 40 meters . Of
the four species known from the world , two species Tachypleus gigas (Muller) and Carcinoscorpius
rotundicauda (Latreille) are known from the eastern cost of the Indian region with a distribution from
Bay of Bengal to Malay archipelago or upto Philippines . Thease primitive animals have retained many of
their interesting characters of related but extinct invertebrates e.g. , trilobites and eurypteryds and
because of their morphological similarities with the fossil , Mesolimulus walchi (Stormer, 1952) they
are called "living fossil" .
Although horseshoe "crabs" present the arthropodal crab-like features , they are really not at all
crabs , rather a close relative of Arachnida and thereby known as a "phylogenetic relic" (Selander et. al. ,
1970 ; Riska , 1981). After latter half of the paleozoic era, horseshoe crabs showed morpholical features
similar to those of present day species ( Fisher,1982,1984; Sekiguchi,1988).

The horseshoe crabs have been exploited commercially by animal traders and quack medical
practitioners (pic.at p.19), rural people in costal areas sell them as ornamental decorative pieces and
many others . As a result , these animals have decreased considerably in number and completely disap-
peared from some of the earlier known areas of their occurrence .

Despite the wide distribution of Asian horseshoe crabs , very little is known on the biology and
natural history of the animals except the works of Japanese scientists (Sekiguchi , 1988) . little work has
been done on the Indian horseshoe crabs until recently (Annandale , 1909 ; Roonwal , 1991 ; Rama Rao
and Surya Rao , 1972 ; Debnath ,1985 ; 1987 ; 1991 ; Debnath and Choudhury , 1985 , 1987 , 1988 a,b,c;
1992 a,b; Chatterjee et.al. , 1988 ; Choudhury et. al. , 1991 ). It is generally known that the length of
distribution of L. polyphemus in North America is almost identical with the length of combined
distribution of three Asiatic species (Shuster , 1982 ).

Although a considerable number of local or discrete populations exist in both Tachypleinae and
Limulinae , the number of L . polyphemus has been estimated as 1,000,000 or more in Delaware Bay ,
USA ( Finn et. al. , 1990) . But no such attempt of censusing the horseshoe crabs in Indian Sub-
continent ever made until recently ( Debnath and Choudhury , 1988c) . A greater number of earlier
studies have attracted the scientific world there to select these curious and mysterious animals as the
study materials for morphology , visual system , circulatory system , and more recently on the
biochemistry of hemolymph fluid of the specimens (Shuster , 1978 ; Cohen et . al. 1979 ; Bonaventura
et. al. , 1982 ; Novitsky , 1984 and Sekiguchi , 1988).

The present study is amied at providing a broad base of vital information on occurrence , habitant
requirements , food and feeding behaviour and population dyanamics ultimately leading to a conserva-
tion and management plan both by way of identified protected habitat area with maxiumum natural
population .
 7
HISTORICAL RESUME

Out of only four extant species of horseshoe crabs in the present world , one occurs off the east cost
of North America and the others in the coastal weather of South east Asia . Bacause of its limited
distribution , the animal was recorded in ancient Chinese literatures (two old Chinese books like Wu
Du Fu between 250 and 305 A.D. and Jiang fu between 276 and 324 A.D. and others ; c.f. ; Sekiguchi ,
1988) but not in the European before the discovery of the New World . It is to be noted here that no such
informations are available in ancient Indian literatures so far studied . However , according to Shuster
(1953) , " the horseshoe crabs was well known to North American Indians who sometimes ate them , and
often used to pointed tail to arm the tips of their fish spears . When the Europeans first came to the New
World they were already familiar with the living animal , since the Asiatic species had been imported
earlier from the Far East , as a curiosity , along with commodities of trade ' ............ was at first mistak-
enly identified as a crustacean by Europeans who called it the King crab."

A persual of literature survey on merostomate studies conducted by Shuster (1988) reveals the total
of 911 publications from prior to 1600 A.D. upto 1951 and thereafter , average 159 publications are
added each decade bringing the total ranging between 1360 to 1500 . In recent years , however , most of
the works are concerned with biochemical studies on horseshoe crabs , especially on its lysate . There are
hardly 500 to 800 research publications out of the totals (1360 - 1500) , i.e. , only 50% delivery with the
three different aspects like i) morphology including anatomy , histology and morphometry ii) natural
history studies including ecology and ethology and iii) taxonomy , palaeontology and evolution . While
the rest 50% accounts for biochemical and physiological studies .

The limited scope of the present thesis work permits the author only to cite the following perusal
on literature survey . Anatomy and external morphology of horseshoe crabs were worked out by several
workers like Koons (1883) , Benham (1885) , Gravier (1929 a,b) , Shoji (1929 a, b ; 1932) ,
Hoeven (1938) , Snodgrass (1952) , Shuster (1960 b,1982) , Shuster and Horrel (1966) , Eldredge
(1970) , Sekiguchi et.al. , (1976 ; 1978) , Sekiguchi and Nakamura (1979) , Riska (1981) , Yamamichi
et.al. , (1983) , Lafon (1984) , Hammen (1986) , Debnath and Choudhury (1988b ; 1992a) , Clare
et. al. , (1990) , Debnath (1991) , Shuster et.al. , (1992) .

Studies on reproductive system and developmental biology were conducted by Benham (1984) ,
Kingslay (1892 ; 1893) , Iwanoff (1907;1933) , Baptist (1953) , Kobayshi (1957) , Sekiguchi
(1960 ;1966 ;1970a ,b ;1973) , Brown and Humphries (1971) , Bennett et. al. ,(1972) , Brown (1976) ,
Sekiguchi el.al. , (1976a,b;1982 ; 1988) , Sugita and Sekiguchi (1979;1982 ; 1983 ;1984), Brown and
Clapper (1980 ; 1981) . Whereas the anatomy of general digestive system ,its histology , feeding and
digestive physiology have been studied by Schlottke (1934a,b and 1935) , Shuster (1948) , Smith and
Chin (1951) , Smith (1953) , Botton (1981 ; 1982 ;1984a;b;d and e) , Tanimoto and Kondo (1982) ,
Botton and Haskin (1984) , Debnath and Choudhury (1987) , Yamashaki et.al. , (1988) , Debnath et.al. ,
(1989) .Sharrer (1941) , Bargman (1949) , Enami (1957) , Herman and Preus (1973) , Choudhury et.al. ,
(1991) had worked on the central nervous system and delt with neuroendocrinilogy of horseshoe crabs.
But Barber (1956) , Barber and Hayes (1963) , Kaplan et.al. , (1976) and Wyse (1971) had worked out
the functions of various chemo-as well as mechanoreceptors .

The natural history of horseshoe crabs specially Limulus polyphemus including its ecology , life
cycle and different behavioural aspects were conducted by various workers like Shoji (1927) , Goto
 8

and Hattori (1929) , Asano (1942) , Roonwal (1944) , Dunton (1953) , Shuster (1953 ,1954 , 1958 ,
1960a ; 1982) , Baptist et.al. ,( 1977 , 1982 , 1984 , 1985 , 1986 and 1987) , Sekiguchi and Nakamura
(1977 ;1979) , Ireland and Barlow (1978) , French et.al. , (1978) , Rudloe (1978 ; 1979a,b ; 1980, 1981,
1982 , 1983) , Gomez - Aguirre (1979) , Rudloe and Rudloe (1981) , Barlow ( 1982 , 1983) , Cohen and
Brockman (1983) , Chamberlin and Barlow (1984) , Debnath and Choudhury (1985 ; 1988b,c) , Botton
and Loveland ( 1987 ) , Botton et.al. , (1988) , Brockman (1990) , Finn et.al. , (1990) , Ross - Farhang
et.al. ,(1990) .

Molting and other physiological works on Merostomata were conducted by a number of researchers
like Fraenkel (1960) , Banner and Stephens (1966) , Robertson (1970) , Jegla et.al. ,(1972) , Page
(1973) , Johansen and Petersen (1977) , Neff and Gaim (1977) , Laughlin and Neff (1977) , laughin
(1981;1982) , Thys (1984), Sugita et.al. ,(1985) , McManus and Ricci (1989) . Molting and ageing
(growth) , these two important aspects had been worked out by Bolau (1879) , Shuster (1954 ;1982) ,
Ropes (1961) , Shuster and Horcell (1966) , Jegla and Costlow (1970 ;1979a,b) Jegla (1972) . While ,
Bang (1956;1979) , Groff and Leibovitz (1982) ,Leibovitz (1986) , Leibovitz and Lewbart (1987) , Miyata
et.al. , (1989) , Kawano et.al. , (1990) , Muta et.al. , (1990) were studied on different aspects of the
disease and immunity problems .

The geographic distribution of horseshoe crabs was studied by a number of workers e.g. Sewell
(1912) , Waterman (1953 ; 1954) , Beamont and Mansueti (1955) , Shuster ( 1955 ; 1958 ; 1979 ;1985),
Teale (1957) , Sekiguchi (1978) , Sokoloff (1978) , Sekiguchi and Nakamura (1980) , Sellenschlo
(1983), Botton and Haskin (1984) , Shuster and Botton (1985) , Botton and Ropes (1987) , Debnath and
Choudhury (1989) . Predation on horseshoe crabs was reported by Rudloe and Rudloe (1981) , Shuster
(1982) , Botton (1984c) , Debnath and Choudhury (1988a)

In recent years the most promissing field of research works on hoeseshoe crab's "blood" espacially
its lysate may be reflected by the benefit of human being : Dores and Herman (1980), Pearson and
Weary (1980) , Bishayee et.al. , (1980) , Dorai et.al. , (1981) , Srimal et.al. , (1985) , Roa and Bhagirathi
(1989) and many others .

Palaeontological works including phylogeny and evolution of Xiphosura , mainly , were of lves
(1891) , Stormer (1952 ; 1955) , Selander and Yang (1970) , Selander et.al. , (1979) , Barthel (1974) ,
Eldredge (1974) , Fisher (1975 ; 1977 , 1982 ; 1984) , Sekiguchi and Sugita (1980) , Miller (1982) ,
Shishikura et.al. , (1982) , Sugita et.al. , (1982) , Miyazaki et.al. , (1989) .

Literatures exclusively on horseshoe crabs have been published by several workers (Cohen et.al. ,
1979 ; Bonaventura et.al. , 1982 ; and Sekiguchi , 1988) which shed light on better understanding of its
biomedical , physiological and environmental research and also on general biology . Number of popular
articles and monograph works also appeared time to time elucidating its importance to the welfare of
human being [ Nishi (1975) ; Rudloe and Rudloe , (1981) ; Novtsky (1984) etc.]

In fine , regarding the taxonomic position of horseshoe crabs , Linaeus (1758) , Muller (1785) ,
Latreille (1802) , Leach (1819) had given a shape of taxonomic position in their respective books of
classification . But the most important contributions to the modern taxonomy were van der Hoeven
(1838) , Dana (1852) , Willemoes-Suhn (1883) , Heymons (1901) , Pocock (1902) . Accordingly, the
existing systematic position of hoeseshoe crabs is accepted as :
 9

Phylum : Arthropoda
Sub-phylum : Chelicerata Heymons , 1901
Class : Merostomata Dana , 1852
Sub - class : Xiphosura Latreille , 1802
Order : Xiphosura Latreille , 1802
Sub - order : Limulina Richter and Richter , 1929
Super family : Limuroidea Leach ,1819
Family : Limuroidea Leach ,1819
Sub-family : Limulinae Leach ,1819
Genus : Limulus Muller ,1785
Species : L. polyphemus (Linnaeus ,1785)
Sub-family : Tachypleinae Pocock , 1902
Genus : Tachypleus Leach , 1819
Species : T. gigas (Muller ,1902)
Species : T. tridentatus (Leach ,1819)
Genus : Carcinosoorpius Pocock , 1902
Species : C. rotundicauda (Latreille ,1802)

But the Japanese workers (like, Sekiguchi , 1988) have proposed for a new taxonomic system in the
family Limulidae considering all sorts of biological works of horseshoe crabs i.e. , comparative anatomy,
hybridization (cross breeding) , biochemistry , genetic studies and others . According to them the three
Asian horseshoe crabs belong to the same genus , Tachypleus . Proposal for a new taxonomic position
thus stands as follows :
Family Limulidae
Sub-family Limulinae Leach,1819
Genus Limulus Muller,1785
Species : L. polyphemus (Linnaeus ,1785)
Sub-family Tachypleinae Pocock,1902
Genus Tachypleus Leach,1819
Species T. tridentatus (Leach ,1819)
Species T. gigas (Muller ,1785)
Species T. rotundicauda (Latreille ,1802) . n. Comb.

A Prehistoric progeny
on the exposed sand
flat, whose status is in
RDB and soon be
on..........CANVAS !
And, its fate is then,
really depend
on....................US !
 10
PHYSIOGRAPHY OF THE STUDY AREAS

The eastern , western and southern cost of the peninsular India is ornamented by majestic rivers
having extensive and highly productive estuarine areas . India's most of the largest rivers like Ganga ,
Subarnarekha , Mahanadi and Godavari etc. carry a vast amount freshwater to the Bay of Bengal passing
through West Bengal , Orissa and Andra Pradesh , the three south-eastern costal states respectively . The
Hooghly -Matla estuarine complex , marked as an important spot on the global map for its famous deltaic
Sundarbans and luxuriant mangrove vegetation , is the richest and biggest in the world .This environmen-
tal complex supports a galaxy of benthic fauna of which horseshoe crabs are most conspicuous .
o o o
The Indian part of Sundarbans is located geographically between 21 to 22 30`N latitudes and 88 to
o
89 29`E longitude . Its costal zone is in the form of low , swampy terrain characterized by extensive
fluvio-marine plains with network pattern of creeks and parallel beach ridges with intervening estuaries
and mudflats (Mangrove Report , 1987) .

The total coast line from Sundarbans to Baitarani delta (Fig. 1& 2) of about 300 km , is inturruped by
large estuaries , Hugli (= Hoogly) , Matla , Saptamukhi and Gosaba in 24 parganas , Rasulpur river in
Midnapue of West Bengal , Subarnarekha , Burhabalaga , Baitarani and Mahanadi rivers in coastal
Orissa . The eastern coast of West Bengal is characterized by sand and mud bottoms , while the coastal
plains of Midnapur in West Bengal and Balasore , Cuttack and Berhampur districts of Orissa are charac-
terized by the sandy shores mainly . But from the Godavari river mouth towards the south-eastern
part of Andra its cost is characterized mostly field areas in regard to horseshoe crab studies is given
below (Fig.1b):

A) Prentice island :
o
It is an elongated and narrow , slightly bent , virgin island ; situated between the latitude 21 43` to
o o o
21 46`N and the longitude 88 18` to 88 19` E in Sundarbans . This mangrove island is surrounded on all
sides with the river Saptamukhi that has a link with Moorigonga via the Hatania - Doania Khal dividing
Namkhana into two halves on the west . On the east and south east , there are two islands : Pathar Pratima
containing a National Crocodile Project (established in 1982) at Bhagabatpur and the Luthian island with
a bird sanctuary , respectively . Prentice island with its rich but shorter mangrove vegetations and numer-
ous creeks and inlets provides a model ecosystem for the study of various micro-and macro-fauna
including the horseshoe crab , Carcinoscorpius rotundicauda .

B) Sagar island :

The largest delta in the western sector of Sundarbans , is situated nearly 85 km south of Calcutta in
o o o o
deltaic West Bengal . The island extends from 21 38` to 21 57`N latitude and from 88 2.35` to 88 11` E
longitude and covers an area of about 235 km2 . This is a tide-dominated island where the tidal range varies
from 5 to 6 mt. and the island is only 6.5 mts. above the sea level (Mukherjee , 1983). It is sorrounded by
a large bodies of water , river Hoogly in the north and western side and river Mooriganga in the eastern
side . The southern part of the island faces the open sea , Bay of Bangal . It has 12 tidal creeks amongst
which the Chemaguri creek has a bound mud-flat enriched with artificial mangroves that support the
lixuriant population of C . rotundicauda as the breeding ground .
 11

C) Digha :

A prominent tourist place of Kanthi coastal plain of Midnapur district directly facing the Bay of
Bengal . The micro-physiographic features of Digha are i) beach and runnel (channel) topography in the
foreshore regions , ii) dunes and sand flats and, iii) tridal creeks , silt flat and marshy lands of east and
west .

The diurnal type of tridal action (two hige tides and low tides) wash the beach everyday with a cyclic
of erosion and deposition . During the high tides the rolling waves transport maximum sediments and
deposited in the back shore areas . The long and wide sea beach (slope measured 1.0 - 1.50 ) is interrupted
by small and narrow tridal runnels by which tridal water enters the dune flats . Dunes are migrating in
nature and to check the northern migration of the dunes the Government organaizations like Department
of Irrigation and Forest planted Casuarina equisetifolia in rows , on the dune furface . Near the
Subarnarekha delta and in the mouth of Talshari Khal , tridal creeks , marshy lands , mud and salt flats , and
terraces are the important morphological features .

The growing need of tourism causes serve human interference in to the beach and overfishing all
among the shore line prevents the migration of Tachypleus gigas to its breeding ground situated at the
shore and at present the natural population is so reduced that for collection of fewer horseshoe crabs one
has to depend on the fishing net .

D) Chandipur :

A recently growing se resort , situated in Balasore district , about 10 kms. away from the township .
The surface of the Balasore costal plain is more or less terraced , the lowest terrace among the sea margin
being impregnated with salt where the present Chandipur is situated . Probably the sea beach of Chandipur
is almost flat one and widest (about 4 kms.) in the India with only 0.4 to 0.50 slope . Foredunes are
developed on the margin of highest high water level . Water mass remains on the beach behind the sand
bars even after receding of water during ebb tide .
To the north-east of Chandipur , a short and narrow estuary , called the Burhabalange , is situated
(Fig.2) . the estuary gradually becoming narrower the shore line was straightened and the river bank
shifted south-west ; the successive positions of shift are marked by sand ridges of indefinite origin and
small scraps . The mouth is marked by huge sand bars . Chandipur sesshore harbours a considerable popu-
lation of Tachypleus gigas .

E) Dhamra :
This estuary is an unique mangrove ecosystem , situated at the junctional complex between rivers
Baitarani and Brahmani in costal Balasore of Orissa . The study area was at the north-eastern part of the
main estuary . Wide mangrove land and mud-flat extended upto 5000 mts. down to the littoral zone . Slope
of the beach was found 0.90 at every 1000 mts. (measured by Clinometer) i.e. the total slope , thus , stands
4.5 to 50 for the entire beach extended upto the lowest tide mark . Between infra- and supra-littoral
zones , comparatively shorter mangrove plants , grasses etc. grow . This is the only place that that harbours
both C. rotundicauda and T. gigas in the inlet creeks or on the mud flat .
About 280 km. long coast line of Balasore , Midnapur and 24 Parganas (deltaic Sundarbans) are
being eroded in different rates in different seasons and the rate of erosion is estimated to be higher in
Digha , Junput , Sagar Island and Bakkhali than in the Balasore coast line (Paul , 1986)
 12
HYDROLOGY

The aquatic environment of estuarine Sundarbans appeared to be more or less stable during January
to June , when the salinity was highly fluctuating ranged from 8.2 to 28%0 as compared to the water
temperature 20 to 340C ( Mangrove Report , 1987) .
On the other hand , the sea shores of Digha and Chandipur , if compared hydrologically , featured
an almost simillar aquatic environment where the salinity ranged from 19.0 to 31.0%0 with the tempera-
ture ranging from 24 to 350C . But in estuarine Dhamra and Mahanadi delta near Paradip port the salinity
ranged between 10.5 to 29.0 %0 with the range of water temperature varied from 23 to 340c (Source :
Marine Survey Deptt., Paradip Port Trust , 1985) ( Fig.1a.)

Fig.1a.
50 Hydroclimatograph Envelopes of the extremes in the salinity / temperature
environment during the months of January through June at three coastal
areas of India:
o
1. Estuarine Sundarbans (Mangrove Reports,1987) : 8.2 - 28.0 ppt & 20 - 34 C
2. Etuarine Dhamra and the Mahanadi Delta near
o
40 Paradip Port ( MarineSurveyDeptt.PPT,1985 ) :10.6 - 29.0 ppt & 23 - 34 C
o
3. Sea shores of Digha & Chandipur :19.0 - 31.0 ppt & 24 - 35 C.
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20
TEMPERA

10

0
10 20 30 40

SALINITY( ppt )

( Courtsey : Prof. Carl N. Shuster, Jr., 1993 )

 13

Fig . 1b.

Shows the coastal West Bengal (the Kanthi coastal plain

west to the River Hugli = Hoogly and the estuarine
Surdarbans to the east) with three study areas (A, B & C) : A.
Prentice island , dots represent collecting sites, B. Chemaguri
creek mouth (Sagar Island) and C. Digha sea shore .

Scale
1:1,000,000

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Fig . 2.

Shows the coastal Orissa with two study sites,where

sampling was done ( A & B). A. The Chandipur sea
shore (District Balasore) and B. Koith-kola mud
flat at estuarine Dhamra (District Cuttack,).
 15
MATERIALS AND METHODS:
PART I : STUDIES ON POPULATION ASPECTS
A. Geographic distribution and habitat :
1. Areas surveyed :

In search of horseshoe crabs during breeding season in the maritime states of Orissa specially at the
coastal districts like Puri , Cuttack and Balasore , regorous field trips were conducted during the period ,
February to March , 1985 . Two more short surveys were conducted in May (at Chandipur , district
Balasore) and June and July (at digha , Junput and Subarnarekha river delta) . Besides those , local tours
were also conducted at every fullmoon tides at Sagar Island , Frazerganj , Canning and Gosaba regions of
Hoogly-Matla estuarine complex . Every time , the author with field assistants visited lonely coasts ,
fishing villages , harbours or ports and tried to get reliable informations . In the next year (1986) , half
yearly tours were also made to Prentice island (W.B.) successively from March , 1986 to August 1987 .
The survey was also made at Dhamra (Koithkhola , district Balasore) twice in May ,1987 .

2. Methods of survey :

Horseshoe crabs were found to breed on the littoral sand or mud beaches indiscriminately as
discrete and isolated populations . Thus it was very tough to establish a particular population located
during survey . However , two or three such specific fields were established where the horseshoe crabs
found to venture regularly at spawning or breeding season . Local men called the "crabs", Ram-Lakhsman
or simply Laxsmania in costal Orissa , while in West Bengal these bizzare animals popularly termed as
Rajkankra (a Bengali word meaning "King-crab") . Living or dead specimens were collected and tried
to confirm by asking or consulting local people , for the situation , if the specimens had actually inhabited
there or had come from other places accidentally . When it was ascertained that horseshoe crabs were
actually living in the area , and effort had been made to find eggs deposited on the sea shore by digging
the mud or sand .

B. Morphology and Morphometry :

Total of 150 T. gigas (100 male and 50 female ) and 75 C. rotundicauda (39 male and 36
female) were collected from various field stations of costal Orissa and West Bengal for studying the
comparative body weight , size and shape . eleven parameters including body weight , such as inter-ocular
length (IOL) , dorsal prosoma breadth (DPB) , dorsal body length ( DBL) ,excluding the telson , ventral
prosoma breadth (VPB) , telson length (TL) , its breadth (Tb) and heigth (FMH) , length of the moveable
marginal spines (MSL) were considered for the study . All morphometric variables were expressed in
nearest mm. except body weight (gm) (Fig. 17B)
In a separate attempt , 23 mated pairs of T. gigas were collected on 8th March , 1985 from
Chandipur sea-shore during the fullmoon high tide , between 10 AM to 12 N. The animals were readily
segregated and the aforesaid parameters of individual males and females were recorded within three
hours of collection . In this study , nine paired characters ( e.g. IOL , DBP, DBL , VPB , TL , CH ,
frontal length i.e. FL , FMH and RL as the distance between median tip of the prosoma and the base
of chilaria) and two unpaired characters , e.g. PL (male clasper length) and Sl (the distance between
dorsal opisthosomatic margin of female and prosomatic margin of male ) were considered for
biometric analysis . Besisde the characters mentioned above , the total body weight and total egg -mass
(EM) were also taken into consideration . In this context , it should be mentioned that random sampling
 16

were done for measuring egg-diameter and calculating total egg-numbers . The length-weight relation-
ship study was based on the data obtained from the total of 45 living C. rotundicauda comprising 28
males and 17 females collected from Prentice island during the period from February to September ,
1986 . For the similar study , total 104 live horseshoe crabs belonging to the species T. gigas comprising
85 males and 19 females were collected from Digha sea shore , during the period of June to July , 1985
. In all the cases , total body length of the animals was measured from the tip of the cephalothorax over
the mid-dorsal line down to the opisthosomatic end and the telson was excluded (expressed in cm.). All
measurements were taken with standard scale or measuring tape , while the weights were taken with an
automatic pan balance (Yamato , Japan) . Collections were restricted only to the breeding population
which came ashore to spawn in the spring and summer months , ranging from March to July of 1985 to
1987 .
Statistical methods like simple and partial correlations , ANCOVA , 't'-test , Chi-square test etc.
used for morphology and biometric analyses were based on Bailey (1959) , Lewis (1966) and Snedecar
and Cochran (1967) . The length-weight study was based on the basic principle of Le Cren (1951).

C. Absolute population estimates :

1. Study areas :
Chandipur (district Balasore , Orissa) and Digha (district Midnapur , West Bengal) ,two well
known sea shores are recognised for horseshoe crabs' breeding and abundance (Roonwal , 1944 ;
Panikkar ,1951 ; RamaRao and Surya Rao , 1972 and Debnath , 1985) . These places were selected for
making-recapture studies . The two shores are regularly inundated by the high saline water (19.0 to 31.0
ppt.) of Bay of Bangal and regarded as the spawning ground for one of the two horseshoe crabs i.e. , T.
gigas .Both these sea resorts receive fresh water from the river , Burabalanga at Chandipur and Talsheri
Khal (Digha Khal) , and Subarnarekha at Digha (Fig. 1,2) . Horseshoe crabs were found to come at shores
during fullmoon high tide of spring and summer months at Chandipur . But their inshore migration was
interfered by 10-15 long tow nets each handled by 30-40 fishermen everyday at Digha coast . 'Crabs'
collected from the nets were usually considered for making the present study .

2. Individual marking :

Individual marking by mechanical means was done rather than the use of levels , tags , paints
or dyes . The animals captured in pair or solitary were gently marked with steel-needle on their carapaces
(dorso-lateral surface) as code numbers like 7/3-1 , 7/3-2, 7/3-3, ......... 26/5-1 etc. (date/month-sl.no.) .
Both the male and female of the same mating pair were marked with similar codes (Fig. 19A, C).

3. Handling and release :

Handling of horseshoe crabs is very much difficult; and precautionary measure was taken by
wearing gloves in the hand . At Digha , total net collections of the day were taken and marked with code
numbers on the dorsal-lateral surface of the animal carapaces . At Chandipur handling of horseshoe crabs
was rather more difficult . Sufficient care was taken at the time of capture and making so that the horse-
shoe crabs felt minimum disturbance and they were released at the end of each operation during low tide
at discrete locations . The horseshoe crabs damaged by predator crows , Corvus splendens L. ( Debnath
and Choudhury , 1988a) or heavily infested with various of epifauna were removed from the primary
capture-stock and these were rejected .
 17

4. Line-transect method for population estimation :

Number of T. gigas was counted in eight random transects measuring 10 X 10 sq. mt.
within an area of 1 sq. km. at Chandipur sea shore (Table 18). Such studies were conducted from 20th to
31st March of 1986 and the total population was calculated following usual line transect method
(Andrewartha , 1970) and the figures were expressed in two ways : i) in terms of days and ii) in terms of
area (toa) .

5. Population estimation by Regression methods :

The results of estimated population (N) , number ofcaptured (C) and recaptured (R) animals
derived from Jackson's method (1939)for Chandipur sea shore (March to July , 1985) and Jolly-Seber's
method (1965) for Digha sea shore were transformed into logarithms . Finally , the transformed values of
N , C and R were correlated and regressed by the following statistical methods of Lewis (1966) and
Bailey (1959) .

D. Breeding activity ; sex-ratio and predation :

Breeding activity was observed in the feilds . Mechanism of pairing (making amplexus i.e. ,
grasping the famale's opisthosomatic marginal spines with the modified clasper legs of the male ) ,
movement , burrowing and dispersal activities were noticed in both C. rotundicauda and T. gigas at five
selected field stations of Prentice island , Chemaguri mud-flat (Sagar Island) , Digha sea shore , Chandipur
sea shore and Koith-kola (Dhamra mangroves). Total males and females were counted to determine the
sex-rations . Marking with steel-needle and tagging with colour rubber bands (in the telson-trunk) were
done in both species to demonstrate the spawning movement along the Bay shores .
Active predation of the horseshoe crabs by crows, Corvus splendens was an accidental obser-
vation during the study to the breeding activities of T.gigas . In order to study the effect of predation on
the sex-balance , capture-recapture method was employed at Chandipur (in addition to get the population
estimate) . On many occasions dead or moribund 'crabs' , especially the females , were found long after
the days of release . After the capture - recapture method has been performed the author visted the beach
again and found a huge number of dry carapaces (females mainly) in which some of them bore the previ-
ous needle markings . From the whole data the approximate population size and sex-ratio were obtained .
At Digha , horseshoe crab population did not come across the tidal edge to lay their eggs due to
human interference . They were caught during fishing and the fishermen threw them on sand believing
them as disturbing elements . Crows , thus , got access to the crabs very easily at the exposed beach . Day
to day collection was sorted out to estimate the sex-ratio of the crabs , and at the same time dead crabs
(dry shells) were also collected at random to determine the sex-ratio .

E. Epifaunal Associates :

Ecto-commensals or epifaunal organisms were collected from dorsal as well as ventral surface
of T. gigas and C. rotundicauda in spring and summer months ( from 1985 to 1988 ) at five coastal
stations viz. Prentice island , Sagar Island and Digha of West Bengal ; and Chandipur and Dhamra of
Orissa . Among the ecto-commensals barnacles constituted the major part . Only noticeable epifaunal
organisms (at least a colony of barnacles) were taken into consideration to certify a horseshoe crab as
infested . The ectocommensals were scrapped off the shells of 50 highly infested horseshoe crabs
(32 T. gigas and C. rotundicauda ) and transferred in 70% alcohol for further population study (
 18

PART II :
STUDIES ON FEEDING BEHAVIOUR ;
FOOD , FEEDING AND DIGESTION

A. Gut content analysis :

Seventy two adult horseshoe crabs ( T. gigas) comprising 42 solitary males , 15 mated males
and 15 mated females from net collections at Digha sea shore were considered for gut content analysis
during May 1985 . The 'crabs' were primarily anasthesized within half and hour after collection , by inject-
ing 10 % formalin in the cephalothorax . By next 30 minutes the animals were sacrificed to remove the
entire guts (gastro-intestinal tracts) from the body , preserved in 3.7% sea water formalin for 12 hours (as
the preservative makes the gut hardened) and then transferred to 70% alcohol . Later in the laboratory, the
guts were measured (length , diameter and weight) , and dissected out the gut contents . The cleared guts
after the removal of contents were again weighed . The contents were preserved in small glass-vials con-
taining 70% alcohol . Samples of gut contents were examined under a stereoscope (Olympus) . Counting
of prey items and grouping them upto the lowest possible taxa were done following the Botton's tech-
nique (Botton , 1984a) . The food matters , whether partly or fully recovered , were sketched and drawn
after camera lucida for determination of probable size and shape according to scale (mm.)
In similar way C. rotundicauda obtained from Chemaguri mud-flat , Prentice island and
Dhamra , and T. gigas obtained from Chandipur and Dhamra of Orissa were also sacrified to identify the
food-matters (Fig. 32A) .

B. Digestive physiology in context to prey items :

For quantitative estimation of food ingested , ten mating pairs of T. gigas were considered .
The amplexed pairs were obtained from Digha sea shore in May 1987 and they were readily segregated
and primary preservation in the feild was done by injecting 10% formalin in the ventral part of
cephalothorax . As described above each of the 20 samples (10 male + 10 female) was examined follow-
ing the procedure developed by Botton (1984a) .
Another batch of 10 pairs of T. gigas of almost similar size were chosen for enzymatic study ,
sacrificed after capture , in the laboratory . The gut was flushed with distilled water and three regions of
the digestive tract dissected out-oesophagus , gizzard and stomach intestine , together with the digestive
gland (hepatopanereas). Crude enzyme homogenates were prepared with glass distilled water , in me-
chanical homogenizer at 40c . The resultant supernatant was selected as crude enzyme extract for carrying
out biochemical studies .
Proteolysis (both acidic and alkaline) were measured following Ichishima (1970) using 2%
casein as substrate at pH 2.5 and 8.0, respectively ; esterase action on 0.01 % beta naphthyl laurate at pH
7.4 seen according to Seligman and Nachles (1963) ; cellulolysis using 1% carboxymethyl cellulose at
pH 5.5 as substrate by the method of Kesler and Tulou (1980) ; invertase action on 2.5% sucrose at pH
5.5 following Pal et. al. , (1980) and amylolysis following Bernfeld (1955) using 1% amylum at pH
6.9 .Total water soluble protein content of extract was measured according to Lowry el. al.,(1951) .
 19

PART III : STUDIES ON ANATOMY AND CYTOMORPHOLOGY OF THE

CNS WITH SPECIAL REFERENCE TO ITS
NEUROSECRETORY ELEMENTS

Both sexes of adult horseshoe crabs , C. rotundicauda , were collected from

Chemaguri mud-flat of Sagar Island (West Bengal) during the months of May to June 1988 . Fifteen
animals having carapace width of about 152.0 to 210.0 mm. for 8 males (weigth ranged between 147.0 to
378.0 gm.) and 182.0 to 225.0 mm. for 7 females (weigth ranged between 305.0 to 629.0 gm.) were
sacrificed following the procedure of Lochhead (1950) for both anatomical and cytomorphic studies on
the Central Nervous System (CNS) . Different parts CNS were fixed in aqueous Bouin's fluid . The tissues
after dehydration were embeded in paraffin (56-580C) and serial frontal sections (15 mm.) were treated
with Haeidanhain's Azan (Gurr , 1959) , Chromealum haematoxylin pholixin (modified by Bargmann ,
1949) and Masson's trichrome staining techniques (Gurr , 1959) .

Fig.2a.
Selling boiled juice with
mustard oil extracted
from horseshoe crabs has
been a common practice
in Bengal..........is not this
against T he W ild-lif e
ild-life
( Protection ) Act !!!
 20

PART - I
STUDIES ON POPULATION ASPECTS
 21

A . Geographic distribution and habitat

INTRODUCTION

Of the 4 extant species , the American horseshoe crab , Limuluspolyphemus occur in the
Atlantic coast of North America , widely distributed from Maine to Yucatan (Shuster , 1953 , 1954 , 1957
, 1979) . The other three Asiatic species , Tachypleus tridentatus , T.gigas and Carcinoscorpius
rotundicauda occur in the Indo-Pacific coasts . T.tridentatus is found in western and southern Japan ,
Taiwan , Philippines and north Borneo (Pocock , 1902) . T. gigas and C . rotundicauda are found on the
coast of India , Bangladesh , Myanmar , Thailand , Malaysia , Philippines and Borneo (Annandala , 1909 ;
Choudhury & Hafizuddin , 1980 ; Pocock , 1902 ; Smedly , 1929 ; Sewell , 1912) . T. gigas was also
recorded from Torres Straits (Pocock , 1902) .
An extensive survey report of Sekiguchi and Nakamura (1980) confirms that 1) all the 3
species are found sympatrically in the north-western coast of Borneo , 2) two species , T. gigas and
C.rotundicauda , are found along the southern coast of Borneo , the northern coast of Sumatra , the coast
of either sides of Malay peninsula and that of West Bengal (digha , Canning , Kkdwip , Freserganj and
Junput) and 3) a combined distribution of T. tridentatus & C. rotundicauda was found along the south-
western coast of Philippins & Palwan Islands . Thus the area of distribution of the three Asiatic species is
surrounded by 4 lines drawn from the western part of Japan to West Bengal , from West Bengal to Java ,
from Java to Torres Strait , and from Torres Strait to the western part of Japan (Sekiguchi & Nakamura ,
1980) . However , no precise information about the distribution of Indian horseshoe crabs , T. gigas and
C.rotundicauda is available ( Sekighchi et. al. , 1976 , 1978 , 1979 , 1980) excepting some scattered
reports (Annandale , 1909 , 1922 ; Panikkar , 1951 ; Pocock 1902 ; Rama Rao & Surya Rao , 1972 ;
Roonwal , 1944) .
The present chapter of the dissertation deals with a detailed survey to record the geographic
distribution of each of the two Indian species in their own territory i.e. in the coasts of Bay of Bangal
(Debnath & Choudhury , 1989 ).
 22
Table 1a
Number of animals observed in different lacalities during
the period February to August , 1985 .

Areas surveyed T. gigas Dead Carcinoscorpius rotundicauda Dead

Couple Male Female carapaces Couple Male Female carapaces

* Chilka - 2 1 - - - - -
Astarang 1 1 - - - - - -
Paradip 5 4 - 8 2 1 - -
Jambu 12 3 - 2 - 1 - 4
Dhamra 7 6 - 4 1 3 - -
Chandipur 320 13 - 154 - 1 - 1
Digha 19 112 - 78 1 - - 2
Sagar Island 5 2 - 3 1 - - -
Frazergaunj 1 - - - 3 2 - 1
Canning - - - - 7 1 - 2
Gosaba - - - - 21 9 - 3

Total 374 153 1 249 37 18 - 13

* Reported and collected by a Telegu fisherman of a deep sea fishing trawler at Paradip port .

Table 1b
Number of animals observed in three lacalities during
the period March 1986 to May 1987 .

Areas surveyed Tachypleus gigas Carcinoscorpius rotundicauda

Couple 0Œ Õ Dead Carcs. Couple 0Œ Õ Dead Carcs.

+ +
Koithkola 44 27 5 35 50 16 1 34
(Dhamra,Orissa)

Digha(Kanthi,W.B.) 5 75 3 35 - 2 - -

Prentice Island 2 - - - 10 18 1 3
(South 24 Parganas,
West Bengal)

Total 51 102 8 70 60 36 2 37
 I W E S T B E N G A L

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COMMENT
If one walked or were able to walk the entire coast line, from
Astarng to to around the Sundarbans, obviously, there would
be an appreciable number of horseshoe crabs, at other sites
too. ( Courtsey: Prof. Carl N. Shuster,Jr. 1993)

Fig . 3.
The map showing the geographic distribution of
Indian horseshoe crabs , Tachypleus gigas and
Carcinoscorpius rotundicauda in coastal Orissa
and West Bengal , India .
 24

RESULTS AND DISCUSSION:

Localities of animal distribution and species occurrence are shown in fig.3. The investigator
has shown the sympatric distribution of the two Indian species , T. gigas and C. rotundicauda in the
Indian coastal region in Table 1. The coastal area of Orissa is discontinuously populated with horse-
shoe crabs , the most dominating species being T. gigas and C. rotundicauda , on the other hand , the 2nd
species dominates the Sundarbans and Dhamra estuaries . Like Limulus polyphemus , the Indian horse-
shoe crabs are also "intermittently distributed" i.e. they are not evenly distributed. Here aslso they occur
in or near the major estuarine river mouths situated at coast of Bay of Bengal. The occurrence of horse-
shoe crabs along the coast of Chilka-lake (Puri district) was confirmed by the information and speci-
mens obtained from a fishing trawler at Paradip port . Except this , everywhere in coastal Orissa the
investigator had encountered horseshoe crabs at field spot. For instancce, the author discovered a mating
couple of C. rotundicauda , 8 to 10 inch. below the mud flat of Mahanadi river mouth at Paradip (north-
ern side) on 28th February 1985 . Again two couples of T. gigas were recovered from the Sandy beach
of Mahanadi river mouth (southern part) the following night (not shown in the map). At Chandipur
(district Balasore) more than 300 mating couples were captured in an area of 2 X 3 sq. km. , a flat stretch
of sand bottom . Similarly at Digha , Junput , Sagar Island , Frazerganj , Prentice island , Gosaba and in and
around port Canning the author came across a number of horseshoe crabs C. rotundicauda , in the mixed
population. It was found to predominate both Sundarbans of West Bengal and Dhamra estuary of Orissa
(Table 1) .
Horseshoe crabs are purely marine dwellers periodically venturing the estuaries (Shuster , 1960a).
The notion of Limulus as a marine species has been superceded by further observations (Shuster, 1979)
Furthermore, "while limuli are most often found in the more saline portions of (and on the continental
shelf),, they have euryhaline tendencies ..........." (Shuster,1982, p.33).
It would seem that C. rotundicauda and Limulus may have a different pattern of distribution in
relation to a salinity gradient than do the two species of Tachypleus ( see. the Clmatographs or
Climographs of fig.1a as was constructed and provided by Prof. C.N. Shuster,Jr. The climograph
envelopes can be further refined into salinity / temperature polygons(Shuster,1990).All the 4 extant
species live in the shallow sea bed or a mouth of river with bottom of muddy sands. C. rotundicauda are
found not only in the sea shore but also in the brackish water zone of the upper stream of a river (Sekiguchi
& Nakamura , 1979) . Annandale (1909) recorded C. rotundicauda in the river Ganga near Calcutta 90
miles north from Bay of Bengal . Sekiguchi and Nakamura (1979) collected the same from a river , about
4 km. away from the sea in Thailand . During the present survey along the coastal Orissa , a single male
specimen of C. rotundicauda was collected at Chandbali , river Baitarani , about 60 km. away from the
sea . Usually the fishermen catch hundreds of specimens of C. rotundicauda about 50-100 km.
interior from the estuarine Sundarbans (port Canning) and sell them to the quaks ( Rama Rao & Surya
Rao , 1972 ; Debnath 1987, Fig.2a) . T. gigas, in most cases , was obtained from the hard sandy beach
facing directly to the sea .
There is a question whether T. tridentatus shares the habitat with T. gigas or C.
rotundicauda . It has been found that T. tridentatus shares the habitat with T. gigas in northern Borneo
(Pocock , 1902) , but no record in sharing with C. rotundicauda . Sekiguchi & Nakamura (1979 ,
1980) showed the western limit of the distribution for T. gigas and C. rotundicauda in the Indian
coast , but that of T. tridentatus was not very clear . Our survey report of 1985 to 1987 revealed that T.
tridentatus actually did not occur in Indian coast (Debnath and Choudhury , 1989) . There is a report that
this species was collected in Vietnam (Waterman , 1953) . The present studies confirms that the western
limit of distribution of the two horseshoe crabs upto the coastal zone of West Bengal and Orissa through
 25

Bangladesh (Choudhury & Hafizuddin , 1980 , Debnath & Choudhury 1989 , Sekiguchi & Nakamura ,
1978 , 1980) .The sympatric occurrence and wide range of distribution of three Asiatic species in the
Indo-pacific seas will provide an important clue to the evolution of these three species (Sekiguchi and
Nakamura , 1980) .
Ives (1891) first compared the distribution of 4 extant species , along the Atlantic coast of
North America and Indo-Pacific coastal water , with the fossil specimens of Limulidae in the Triassic ,
Jurassic , Cretaceous and Oligocene beds of Europe and Syria and suggested that the Mesozoic waters
of Europe were the probable centre of dispersal of the ancestors of the present species . Distribution
of the fossil and living species suggests that Limulidae have always been tropicotemperate species ,
restricted almost entirely to the Northern hemisphere and then L. polyphemus migrating westward
and T. tridentatus , T. gigas and C. rotundicauda eastward . Thus none of the 4 extent species were
derived from the other , but from a common European ancestor or separate ancestral species (shuster,
1957 ,1960b,1982). In addition to geographic ranges of comparable extent , the 4 species have so similar
in ecology , morphology and serology that they probably could be accommodated in one genus
(Shuster , 1962 , 1982) .
From ecological point of view , the present day distribution and extent of horseshoe crabs can
be defined chiefly by the water temperatures , salinities and depths of habitat . Temperature appear to be
the limiting factor for the northern ranges of L. polyphemus and T. tridentatus . In the south , tempera-
ture seems less likely as the limiting factor , at least for T. gigas and C. rotundicauda (Shuster ,
1982) . Amongst the three Asiatic species , only C. rotundicauda is found more adapted to a wide range
of salinity than T. tridentatus and T. gigas . This is evidenced by the occurrence of T. gigas mostly at
the sea shores directly facing the high saline sea-water regime and C. rotundicauda in the brackish
water estuaries and river mouths of Subarnarekha , Mahanadi and Baitarani .
 26

B. Morphology and morphometry

INTRODUCTION

Merostomate morphology has long been a fascinating subject for the zoologists and
paleontologists . Most of the morphological works on extant horseshoe crabs , particularly , Limulus
polyphemus , have already been done (Lockwood , 1870 ; Koons , 1883 ; Pomerat , 1933 , Shuster , 1955/
58 ; Cohen and Brockman , 1983 ; Brockman , 1990 and Shuster et.al. 1992) .
Among the three Asiatic species of horseshoe crabs , Tachypleus gigas was studied well
(Pocock , 1902 ; Smedley , 1929 ; Sekiguchi et.al. 1976) . Morphological variation in Carcinoscorpius
rotundicauda has also been worked out as was observed in T. gigas (Sekiguchi et.al. , 1978) . There-
fore , a perusal of literature survey reveales that except for a few scattered reports (Annandale , 1909 ,
Roonwal , 1944; and Rama Rao and Sury Rao , 1972) no morphological work has been done on the
Indian horseshoe crabs untill 1980s (Debnath , 1991 ; Debnath and Choudhury , 1988b , 1992a ; Chatterjee
et.al. , 1988) .
The present chapter deals , moderately , with the morphology , sexual dimorphism and biometry
of breeding populations including length-weight relationship of the two species , T. gigas and C.
rotundicauda .

T. gigas in dorsal view

C. rotundicauda in ventral view

 27

OBSERVATION AND RESULTS

(a) General organization :

In reneral body plan , the structure is covered by a tough chitinous exoskeleton . The figures
show the following divisions of the body :
1. a prosoma or cephalothorax , bearing the appendages around the mouth (Fig. 16B , D and 17A)
2. a mesosoma or pre-abdomen , bearing the genital operculum and the gill flaps ;
3. a metasoma or post abdomen , which is devoid of appendages ; and
4. a telson , which is long and spine like ( Fig. 4&5) .
The mesosoma (2) and metasoma (3) are fused together into an opisthosoma . Six movable
spines on each side of opisthosoma mark the lateral margins of the somites of the mesosoma . The
opisthosoma is jointed both with the prosoma and with the telson . The body is concave below and the
whole carapace is covex dorsally giving a horseshoe shape . [ The mouth is mid ventral in the prosoma ,
between the bases of the walking legs (five)]. there are two groups of ventral appendages : a) Six pairs of
tubular prosomal appendages - the first pair is chelicera and the rest five pairs of walking legs , repre-
senting the appendages of 1st to VIth somites , and b) Six lamellate plate like abdominal appendages , the
first one is comparatively tough , lacks , gills , forming a gential operculum bearing gonopores on its
posterior wall , while the rest five pairs are connected by a median membrane and each support about 150
gill lamellae . These are simply gill lamellae forming book-gills and comparable with the book lungs of
arachnids . Thus , all those 6 abdominal appendages represent the fused segments of VIII to XIII . The
rudimentary chilaria (of segment VII) resemble the prosomal coxae . The pre-oral chelicerae have three
segments , the distal one chelate . The spined gnathobases of the 5 pairs of walking legs are arranged
almost radially around the mouth (Fig. 5 , 17A) .

(b) Species identity :

T. gigas : Both male and female appeared deep greenish in dorsal shell colour , with a
gigas
triangular and dorsally crested telson (Fig. 18C) . In both the sexes the opisthosomatic lateral spines are
6 in number , but all are long and more or less equal in males , while in females the first three are long and
the rest three are short . A movable spur was present on the 4th segment of the sixth prosomatic append-
age which might be for burrowing purpose (Fig. 18B , D) . Inner branches of the gential operculum were
not extended distal to the tips of outer branches (Fig. 18C) . Dorsal surface of either sexes were found
more spiny than C. Rotundicauda. Note that no juvenile females have movable marginal spines on the
opisthosoma, if present more or less of equal size.
C. rrotundicauda
otundicauda : Animals of eigher sexes looked brownish black or deep greenish black
in appearence , with a rounded non crested (i.e. smooth) telson (Fig. 16B) . Lateral opisthosomatic
spines were all short and 6 in number (Fig. 16D) . No movable spur was present on the 4th segment of the
sixth prosomatic appendage (Fig. 18C) . Inner branches of the genital opercullum were extended distally
upto the tip (Fig. 18C) . Dorsal surface of both sexes were less spiny than T. gigas (Fig. 16C) .

(c) Sexual dimorphism :

T. gigas : The male was comparatively smaller than the females . All the 6 opisthosomatic
spines were long in males , while the first 3 long and the rest three were short in females , fourth and fifth
were being used by the male for clasping . The 2nd and 3rd prosomatic appendages were modified as
clasper organs in males , that is , the terminal portion (tarsus) of the prododite was atrophied and
 28

lost, and only the dactylopodite was present . Thus it looked nonchelate and hook-like (Fig. 18B) . But in
females these two appendages were chelated and looked like normal walking legs (Fig. 18B) .

C. rotundicauda : The male was smaller than the female in comparatively body size . All the
6 opisthosomatic marginal spines were short with the second and third spines slightly longer than the rest
. The 2nd and 3rd prosomal appendages of the male showed well developed terminal portions which were
atrophied in Limulus and Tachypleus . This terminal portion , together with dactylopodite , formed a
chela which looked stouter than the chelae of other appendages (Fig. 18D) . These clasping organs were
conspicuous and black in colour .

(d) Statistical morphology :

Table 2 shows the comparative body weight (wt) , size and shape comprising five major
morphometric measurements e.g. IOL , DPB , DBL , VPB and TL of T. gigas collected from Digha and
Chandipur and C. rotundicauda collected from Prentice and Sagar Islands . Data include population
number (N) , mean value (X) and standard deviation (SD) for each measurement .
Table 3 gives the pool data comprising 100 T. gigas and C. rotundicauda in a comparative
manner . Excluding the body weight , in this table 3 morphometric variables were considered , with only
the range and mean value of respective characteristic . The proportionate means values (i.e. , the male to
female mean ratio are presented in table 4 which will determine the relative size of male to female .
Simple and partial correlation coefficients were worked out considering only four major
morphometric variables e.g. 1 (IOL) , 2 (DPB) , 3 (DBL) and 4 (VPB) , These four measurements were
taken from 90 males plus 19 females of T. gigas and 28 males plus 21 females of C. rotundicauda
(Table 5) . The results of simple and partial correlation computation are presented in table 5 .

(e) Biometry of mating in T. gigas :

A summary of nine paired and two unpaired morphometric measurements , body weight and
weight of the egg mass for the twenty three mated pairs of T. gigas have been presented in Table 6. In
every morphometric consideration the female was considerably larger and in no case did the measure-
ment of the male appear greater than or even closely coincident with a similar female measurement .
Fig. 8 shows graphically that there was some overlapping in the ranges of only two measure-
ments : RL and FMG . Most of the characters , however , were discrete e.g. body weight IOL , DPB ,
VPB , DBL and FL. Only the lower end of the range of measurements for females began at the top end of
the male range so for tail length (TL) and central hight (CH) are concerned . It was also characteristic that
the range of VPB , FL and RL seemed to be equal size , while the range of male TL was greater than the
female range . Greatest variation in range and distance were being displayed by the Wt. class : the female
range was very much higher than the male range .
None of the males of the body dimensions of the sexes overlapped or occurred at the same
point . Further , in no instance did the mean coincides with the mode and all comparisons between
male to female means showed ( t-test) high difference in significant at 0.001 level . Proportion of male
mean to female mean showed 0.720 to 0.821 (average 0.758) , while for weight alone it was 0.349 .
Correlation and regression relationship were computed for mine paired characters (Table-
7). In addition to those relationship between weight , DPB (Male) to SL (Female) and DPB (Male) to
EM (Female) also were computed . Amongst these all , only CH , FMH and RL showed significant
correlation coefficients . In the regression analysis the female measurements assigned along the abcissa
(X) while the male components were placed on the ordinate (Y) . High regression coefficients were
 29

obtained only in the case of FMH (tanè = 25.260, b = 0.472) and RL ( tan è = 33.660, b = 0.662). Since
dorsal prosoma breadth (DPB) and dorsal body length (DBL) have been found to be the most highly
correlated morphometric features in all Indian horseshoe crabs (Debnath 1991) , these two were se-
lected along with body weight (Et.) to arrange the mated animals in tabular forms for chi-square testing
(Table 8) . Theoritical frequencies are shown in parentheses , while the actual ones are on their left . Since
the expected frequencies (in parenthesis) in most cases were found to be less than 5, the 2 x 2 contin-
2
gency X -test could not be done (Baily , 1959) . This was due to a seemingly smaller population (only 23
pairs of T. gigas ) considered for the study . Otherwise the results should bear some significance .

(f) Length - weight relationship :

The study of length-weight relationship provides a mathematical correlation between length
and weight may be converted , one to the other, in future cases where only one of the parameters is
known .The method has been found to be of value in measuring variations in the expected weight or length
of an animal, particularly fish . In fish, such correlations have been used in examining indications of
fatness, gonad development, general well-being and other parameters (Le Cren , 1951) . McMahon
( 1973,1980) has pointed out that this unique morphometric relationship may provide an important bio-
logical information and Peters (1983) suggests that it could led to the formulation of a prospective
biological theory .The application of this method to the Indian horsehoe crabs is explained below ( see
also Fig.7 ) .

Parabolic Equations Logarithmic Equations

T . gigas:
Males : W = 0.03748 L 3.08992 Log W = -1.42611 + 3.08992 Log L .
Females : W = 0.23317 L 2.52090 Log W = -0.63230 + 2.52090 Log L .

C. rotundicauda:
Males : W = 0.4116 7 L 3.09554 Log W = -1.38546 + 3. 09554 Log L .
Females : W = 0.43376 L 2.35312 Log W = -0.36275 + 2.35312 Log L .

Analysis of covariance (ANCOVA) revealed no significant difference (Table -9) between regression
lines of both sexes . So a common regression line for the two sexes obtained from the pooled data as
below :

T. gigas : Log W = -1.66218 + 3.29953 Log L .

C. rotundicauda : Log W = -1.72704 + 3.39719 Log L .

The above common logarithmic equations have got justified values i.e.

F = 0.57839 ; df. 1,100 , P>0.05 for T.gigas

F = 1.23589 ; df. 1,41 , P>0.05 for C. rotundicauda (Table -10)
Therefore , the common parabolic equations for respective species are found as

T. gigas : W = 0.02176 L 3.29953

C. rotundicauda : W = 0.01874 L 3.39719
Table-11 and Table-12 represent the t-test of regression lines and t' -test between regression coeffi-
cients of either sexes , respectively .
Table 2 : Showing weight (gm) and five morphometric variables (mm) of the two species of Indian horseshoe crabs , with population (N) , mean value (X)
and standard deviation ( + S.D.) , collected from 4 stations .

Tachypleus gigas Carcinoscorpius rotundicauda

DIGHA CHANDIPUR PRENTICE ISLAND SAGAR ISLAND

Male Female Male Female Male Female Male Female
N X SD N X SD N X SD N X SD N X SD N X SD N X SD N X SD

WT. 90 175.56 29.95 19 475.26 58.34 30 178.63 31.39 30 511.33 57.88 28 238.75 37.73 2 1 446.71 92.99 18 235.22 34.57 12 456.48 85.34
1. IOL 90 72.81 2.75 19 98.24 5.43 32 72.16 3.92 23 96.71 5.62 28 73.81 4.82 21 88.729 8.14 18 75.68 2.78 12 88.42 3.85
2. DPB 90 165.72 6.18 19 215.28 9.82 32 162.92 6.64 23 219.82 11.84 28 172.35 14.81 2 1 194.51 9.34 18 179.15 11.42 12 200.18 9.83
3. DBL 90 152.18 6.45 19 205.62 8.63 32 150.18 5.08 23 204.65 13.92 28 163.05 9.12 21 186.15 12.64 18 168.34 7.43 12 191.42 11.34
4. VPB 90 144.75 6.09 19 178.13 5.80 30 143.65 6.48 30 179.35 8.21 28 147.84 7.83 21 169.05 11.20 18 153.74 9.68 12 171.67 10.91
5. TL 28 126.39 10.01 19 172.45 11.45 30 128.31 9.62 26 173.18 14.66 24 154.68 11.64 2 0 173.70 11.18 17 153.48 9.34 12 176.74 9.54

IOL , inter-ocular length ; DPB , dorsal prosoma breadth ; DBL , doesal body length ; VPB , ventral prosoma breadth and TL , telson length .

Note:
N stands for the number of individuals measured; IOL = interocular distance ( not length )
and DPB = width or breadth of prosoma measured along the curvature of the carapace, but
not "dorsal prosoma breadth"-----------as suggested by Prof. Carl N. Shuster,Jr. ( 1993 )
30
 31

Table -3 . Showing the comparative body size , shape and weight of two species of Indian horse-
shoe crabs (Except weight in gm all parameters are expressed in mm.

Sl. Variables Tachypleus gigas ( N = 100 ) Carcinoscorpius rotundicauda ( N = 75 )

Male (50) Female (50) Male (39) Female (36)
Range Mean Range Mean Range Mean Range Mean

Wt. (gm.) 120.0 - 225.0 176.36 335.0 - 588.0 478.56 150.0 - 378.0 235.85 305.0 - 629.0 452.21
1. IOL 67.2 - 81.5 71.84 89.3 - 108.5 98.75 61.6 - 82.9 74.24 81.6 -105.2 88.75
2. DPB 155.4 - 181.7 163.85 199.2 - 233.4 216.23 151.2 - 210.6 175.42 182.6 - 224.5 201.45
3. DBL 137.3 - 170.1 153.24 190.1 - 218.3 206.41 141.2 - 191.4 163.52 174.3 - 209.6 188.35
4. VPB 134.8 - 155.5 145.72 168.3 - 189.7 177.15 133.4 - 162.7 148.95 155.7 -192.2 170.88
5. TL 126.5 - 141.8 128.38 154.5 - 190.2 173.12 138.5 - 172.3 155.20 140.5 - 181.3 175.84
6. Tb 5.5 - 6.8 6.14 6.0 - 7.1 6.53 5.2 - 6.1 5.62
7. Th 6.2 - 7.4 6.77 6.8 - 7.6 7.21 5.6 - 6.8 6.18
8. CH 32.4 - 46.5 35.82 39.1 - 55.6 46.52 29.2 - 53.1 37.17 35.4 - 58.6 46.1
9. FMH 10.2 - 15.4 12.55 12.2 - 21.6 18.53 14.2 - 19.4 13.78 13.3 - 22.2 17.55
10. MSL
a) Long 20mm> 20.0 - 23.2 21.81 20.1 - 26.3 22.7
b) Short 20mm< 7.2 - 19.2 16.5 8.7 - 18.9 12.15 5.2 - 11.5 9.32 6.5 - 15.3 11.22

IOL = interocular length , DPB = dorsal prosoma breadth , VPB = ventral prosoma breadth , DBL = dorsal body
length , TL = tail length , Tb & Th = breadth & hieght of tail, respectively , CH = central body height , FMH = frontal
marginal height , MSL = marginal spine length .

Note: Long MSL means greater than 20 mm. and Short MSL means shorter than 20 mm.
 32

Table 4.Showing the ratios of mean values of male to female of two

species of Indian horseshoe crabs .

Sl. Parmeters Tachypleus gigas Carcinoscorpius rotundicauda

Male : Female Ratio Male : Female Ratio
Wt(gm) 176.36/478.56 0.3685 235.85 /452.21 0.5215
1. IOL 71.84 /98.75 0.7274 74.24 /88.75 0.8365
2. DPB 163.85/216.23 0.7577 175.42/201.45 0.8707
3. DBL 153.24/206.41 0.7424 163.52/188.35 0.8681
4. VPB 145.72/177.15 0.8225 148.95/170.88 0.8716
5. TL 128.38/173.12 0.7415 155.20/175.84 0.8826
6. T.b. 6.14 / 6.53 0.9402 - -
7. T.h. 6.77 / 7.21 0.9389 - -
8. CH 35.82 / 46.52 0.7699 37.17 / 46.01 0.8078
9. FMH 12.55 / 22.07 0.5686 13.78 / 17.55 0.7851
10. MSL
i) long 21.81 / 22.07 0.9882 - -
ii)Short 16.05 / 12.15 1.3209 9.32 / 11.22 0.8306

Except Wt & MSL: X = 0.7787 X = 0.8466

SD = 0.1138 SD = + 0.0373

NOTE: IOL = interocular distance ( not length ) and DPB = width or breadth of prosoma measured
along the curvature of the carapace, but not "dorsal prosoma breadth"---------as suggested by
Prof. Carl N. Shuster,Jr. ( 1993 )
 Table 5. 33
Simple and partial correlation coefficients between four morphometric variables,
1(IOL),2(DPB),3(DBL) and 4(VPB) of two species of Indian horseshoe crabs

Tachypleus gigas Carcinoscorpius rotundicauda

Male(df, 89) Female(df, 18) Male(df, 27) Female(df, 20)

Simple correlation coefficient

r
12 0.7185 a 0.7218 a 0.6601 a 0.8560 a
r
13 0.7028 a 0.7162 a 0.7583 a 0.7944 a
r
14 0.3750 a 0.6925 a 0.6575 a 0.9167 a
*r
23 0.8531 a 0.8299 a 0.8878 a 0.8700 a
r
24 0.3505 a 0.7253 a 0.7023 a 0.8544 a
r
34 0.3152 a 0.7637 a 0.6228 a 0.81

1st step partial correlationcoefficient

r
12.3 0.3203 b 0.3272 -0.0439 0.5506
r
12.4 0.6761 a 0.4421 0.4237 a 0.3506
r
13.2 0.2475 b 0.3035 0.2125 0.1948
r
13.4 0.6645 a 0.4020 0.5917 a 0.2156
r14.2 0.1892 0.3546 0.3625 0.6901
r14.3 0.2275 0.3229 0.3631 0.7665
*r
23.1 0.7037 a 0.6479 a 0.7908 a 0.6031
*r
23.4 0.8355 a 0.6211 a 0.8091 a 0.5816
r
24.1 0.1257 0.4514 b 0.5017 a 0.2601
r
24.3 0.1651 0.2338 0.4148 0.5146b
r
34.1 0.0780 0.5318 b 0.5148 a 0.3437
r
34.2 0.0327 0.4213 0.5917a 0.2668

2nd step partial correlation coefficient

r
12.34 0.2960 b 0.2670 -0.1162 0.2835
r
34.12 -0.0155 0.3531 -0.0878 0.1864
*r
23.14 0.7016 a 0.5398 b 0.7647 a 0.5330 b
r
14.23 0.1868 0.2971 0.4195 b 0.6750 a
r
13.24 0.2449 b 0.1816 0.4676 a 0.0172
r
24.13 0.0999 0.1656 0.4628 a 0.3220

where, a , P< 0.01 and b , P< 0.05

 34

Table 6.
Summary of statistical analysis of body weight (wt /gm) and eleven body mesurements of
twenty three mated pairs of T . gigas

Measurements Sex Range X 0 0 SE Mode Mean t value P*

n-1 n Proportion

M 120.0 - 220.0 167.93 25.72 25.15 5.36 150

Wt 0.349 153.703 p
F 355.0 - 585.0 480.87 59.48 58.17 12.40 440

M 67.2 - 78.4 73.26 3.01 2.94 0.62 72

IOL 0.744 129.896 p
F 89.0 - 107.0 98.46 5.44 5.29 1.12 95

M 155.0 - 181.2 166.03 5.67 5.55 1.18 165

DPB 0.771 195.289 p
F 199.3 - 232.0 215.29 9.95 9.24 1.97 212

M 144.5 - 161.3 151.50 4.24 4.15 0.88 150

DBL 0.739 177.508 p
F 190.0 - 217.0 204.93 8.57 8.38 1.78 210

M 135.0 - 157.0 146.11 5.01 4.89 1.04 150

VPB 0.821 127.211 p
F 168.2 - 189.0 178.4 6.23 6.09 1.29 175

M *99.5 - 145.0 128.76 9.92 9.71 2.06 130

TL 0.777 83.681 p
F 149.0 - 181.0 165.58 9.91 9.69 2.06 172

M 27.4 - 29.1 34.18 2.77 2.71 0.57 33

CH 0.735 136.511 p
F 40.0 - 55.0 46.47 4.07 3.98 0.84 45
________________________________________________________________________________________________________________________
M 28.2 - 35.3 32.31 1.59 1.55 0.33 34
FL 0.733 136.511 p
F 39.0 - 47.3 44.05 2.27 2.22 0.47 45

M 10.5 - 15.2 12.62 1.61 1.57 0.33 34

FMH 0.720 43.959 p
F 12.2 - 21.6 17.52 3.17 3.10 0.66 16

M 51.0 - 81.0 70.01 7.37 7.21 1.53 68

RL 0.789 50.590 p
F 73.0 - 104.0 88.13 8.69 8.50 1.80 82

M 20.0 - 30.0 25.43 2.48 2.42 0.57 23

Pl - - -
F - - - - - -

M - - - - - -
SL - - -
F 12.0 - 35.0 27.61 6.27 6.14 1.30 30

EM F 55.0 - 195.0 133.56 38.13 37.29 7.95 135

* P < 0.001

IOL ,interocular length ; DPB , dorsal prosoma breadth ; DBL,dorsal body length ; VPB , ventral prosoma breadth ;TL ,
tail length ; CH ,central height ; FL ,frontal length ; FMH ,frontal marginal height ; RL ,radial length ; PL,clasper length ;
SL , median free part of opisthosoma and EM , weight (gm) of egg-mass .
NOTE: IOL = interocular distance ( not length ) and DPB = width or breadth of prosoma measured
along the curvature of the carapace, but not "dorsal prosoma breadth"---------as suggested by
Prof. Carl N. Shuster,Jr. ( 1993 )
 Table 7.
Correlation and regression values between
Male and Female parameters .

Wt IOL DPB DBL VPB TL CH FL FMH RL DPB(M) DPB(M)

SL(F) EM(F)
b a a
r-value 0.136 0.260 0.366 0.129 0.199 0.123 0.451 -0.221 0.892 0.746 0.140 -0.224
t-value 0.629 1.233 1.802 0.596 0.930 0.567 2.315 1.038 9.041 5.132 0.648 1.053
p(level of 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.001 0.001 0.05 0.05
significance)
b* 0.061 0.151 0.230 0.066 0.167 0.128 0.321 0.161 0.472 0.662 0.132 0.035
tan0 3.49 0 8.58 0 12.95 0 3.77 0 9.48 0 7.29 0 17.79 0 9.14 0 25.26 0 33.66 0 7.52 0 2.01 0

a & b , Significant at 1% and 5% level , respectively ;

b* , Regress Coefficient of female (X) on male (Y) .
35
 36

Table 8.
Chi-Square arrangement of size (A&B) and weight (C) of mated pairs
(expected frequencies within parentheses)

A . Dorsal Prosoma Beadth (DBP) Measurement

FEMALE Small Medium Large TOTAL

199-210 211-222 223-232

Large 0.(078) 2(0.86) 0.(0.34) 2

173-181
Medium 5(4.69) 4(5.21) 3(2.08) 12
MALE

164-172
Small 4(3.52) 4(3.91) 1(1.56) 9
TOTAL 9 10 4 23

B . Dorsal Body Length (DBL) Measurement .

FEMALE Small Medium Large TOTAL

190-198 199-208 209-217

Large 0.(0.78) 0(0.91) 3(1.30) 3

157-161
Medium 2(2.61) 3(3.04) 5(4.34) 10
MALE

151-156
Small 4(2.61) 4(3.04) 2(4.34) 10
156-150
TOTAL 6 7 10 23

C. Body Weight (Wt in gm)

FEMALE Less Medium Heavy TOTAL

355-430 431-505 506-585

Heavy 1(1.82) 2(2.08) 3(2.08) 6

186-220
Meduim 1(1.82) 3(2.08) 2(2.08) 6
MALE

156-185
Less 5(3.34) 3(3.82) 3(3.82) 11
120-155
TOTAL 7 8 8 23
 37

Table 9.
Statistics (log-trans formed) of the Length Weigth Relationship of
Males and Females of two Indian horseshoe crabs .

SPECIES SEX N SX SY SX2 SY2 SXY

MALE 85 100.42927 190.00707 118.68307 425.23588 224.57082

T. gigas FEMALE 19 24.94337 50.79693 32.75238 135.86436 66.70305

------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

C. rotundicauda MALE 28 33.90302 66.15546 41.08804 156.86390 80.21862

FEMALE 17 21.58707 44.63033 27.43822 117.39868 56.73484

SX2 , SY2 and SXY , sum of squares and products ; SX and SY , sum of logarithmic
values of length and weight , respectively .
Table - 10. Comparison of Regression Lines of Weight on Length of adult male and female of Indian Horseshoe Crabs .

Deviations from regressions

2 2
df Sx Sxy Sy b df SS MS F
Within
MALE 84 0.02379 0.07351 0.49839 3.08995 83 0.27124 0.00326
FEMALE 18 0.00650 0.01638 0.05761 2.52090 17 0.01633 0.00096
(P) POOLED

T. gigas
(M+F) 102 0.03029 0.08989 0.55600 2.96764 100 0.28757 0.00287
Difference between slopes 1 0.00166 0.00166 0.57843
Between P > 0.05
T&P 1 0.26768 0.89327 3.74522
(T) Total 103 0.29797 0.98316 4.30122 3.29952 102 1.05725 0.01036
Difference between adjusted means 1 0.76802 0.76802

Comparison of slopes : F = 0.00164 = 0.57843 (df = 1,100) , P > 0.05 no significant difference
0.00287

Within
MALE 27 0.03751 0.11612 0.55871 3.09554 26 0.19923 0.00766
FEMALE 16 0.02634 0.06199 0.23002 2.35312 15 0.08412 0.00560
(P)POOLED
(M+F) 43 0.6385 0.17811 0.78873 2.78950 41 0.28335 0.00691
42 0.29189 0.00694
Difference between slopes 1 0.00854 0.00854 1.23589
Between

C. rotundicauda
T&P 1 0.03683 0.16392 0.72955 P > 0.05
(T)Total 44 0.10068 0.34203 1.51828 3.397719 43 0.35633
Difference between adjusted means 1 0.06444 0.06444

Comparison of slopes : 0.00854 ( df = 1,41) , p > 0.05 no significant difference

F= = 1.23589
0.00691
38
 39

Table 11.
t-test of regression lines of Males and Females of two
Indian Horseshoe Crabs .

SPECIES SEX(df) SS MS MS Sb t p
T. gigas
MALE(83) 0.27124 0.00326 0.05716 0.37063 8.33702 0.05

FEMALE(17)0.01633 0.00096 0.00309 0.38430 6.55971 0.05

--------------------------------------------------------------------------------
C. rotundicauda
MALE(26) 0.19923 0.00766 0.08752 0.45189 6.85020 0.05

FEMALE(15) 0.08412 0.00560 0.07483 0.46109 5.10335 0.05

SS , sum of squares of deviations from regression ; MS , mean square deviations

from regression ; MS , sample standard deviation from regression ; Sb , sample
standard deviation of the regression coefficient .
 Table 12.
t'-test between regression coefficients of Males and Females of
two Indian Horseshoe Crabs

2
SEX df SX MS b Estimated t' df p = 0.05 t W Expected t'

MALE 83 0.02379 0.00326 3.08995 84 1.987 0.0003835

T.gigas 1.46214 2.05118
FEMALE 17 0.00650 0.00096 2.52090 P 0.05 18 2.101 0.00050521
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

MALE 26 0.03751 0.00766 3.09554 27 2.052 0.0002735

C. rotundi
cauda 1.14990 2.0891
FEMALE 15 0.02634 0.00560 2.35312 P 0.05 16 2.120 0.0003294
40
 Flg 41
PROSOMA
Ocs Doc
Opr
Ecr
Prs
Lcy
Sos Ops
Ldr
Prk
Adm Cdr
Pop pos
Pcs
Atr Ptr
Acf Msa
Arp Enp
Klo
Ldr
Tvn Psa
Lms
OPISTHOSOMA
Sms
Dly
Ana
Dlp

TELSON
Dls

30 mm

Tachypleus gigas
( Hand drawn Dorsal View )

Fig.4 Body structure of the horseshoe crab,T. gigas (female):External anatomy

dorsal view (after Carl N. Shuster,1982 and K. Sekiguchi,1988)

Acf,arcuate furrow; Adm,arthrodial membrane; Ana,anal angle ; Arp,auriculate

process ; Atr, anterior ridge; Cdr, cardiac region; Dlp, dorsal process; Dls,
dorsal serration ; Dly, dorsal yoke; Doc, dorsal ocelli ; Ecr, extra cardiac
region ; Enp, entapophyseal pit; Flg, flange ; Klo, opisthosomatic keel; Lcy ,
lateral compound eye;Ldr, longitudinal furrow; Msa, muscular attachment; Ocs ,
Ocellar spine; Opr,opthalmic ridge; Ops, opthalmic spine; Pcs, Post-cardiac
spine; Pos,Post-opthalmic spine; Prk, prosomatic keel; Prs, pre - opthalmic
spine; Psa, posterior angle; Ptr, posterior ridge; Sms, short moveable spine ;
Sos, sub-opthalmic region; Tvn, transverse nodule .
 42
Gnv
Flg
Dbr Sfa

Mls

Wkl Vso
Chl
Ept
Mth
Pms
Chi
Prp
Flb
Evs
Ivl
Spo
Gol
Mrp

Pts

Bra Anl
Trm
Mvs
Tlt
Tvr

Alr

Vlf

ig.. 5. Body structure of the horseshoe crab , T. gigas (female) :

Fig
External anatomy / ventral view (after Prof. Carl N. Shuster,Jr. 1982 and
K . Sekigchi , 1988) .
Alr , axial area ; Anl , anal slit ; Bra , branchial appendages ; Chi , chilarium; Chl , chelicera ;
Dbr , doublure rib ; Ept , epistome ; Evs , exuvial suture ; Flb , flabellum ; Flg , flange ; Gol ,
genital operculm ; Ivl , interados of vault; Mls , mesial spine ; Mrp, marginal process ; Mth ,
mouth ; Mvs , moveable spine ; Pms , promeso-sternite ; Pts , posterior slope ; Sfa , sub-frontal
area ; Spo , spatulate organ ; Tlt , telson trunk ; Trm , terminal membrane ; Tvr , transverse
ridge ; Vlf , ventral longitudinal furrow of telson ; Vso , ventral sense organ ; Prp, propodite.
 43
Doc

FL
Chl Vso

PROSOMA
IOL Ovd

Mth
DPB Gnb

VPB Chi EM

OPISTHOSOMA
SL
Ovd

Gol

Gop
Mts
Anl
Marginal spines

Dorsal serration
Anl

B .
TELSON

B . Ventr al vie
entral vieww
A . Dorsal view
A .
(showing left half of
reproductive system)
Dorsal hinge
DBL
ƒ
FMH

Movable spine
Lcy
CH

BL TL
VBL

C . Lateral view DPB

Doc
CH

FMH

VPB
D. Fr ontal vie
vieww

Fig.6.
Three Dimensions of a Female Tachypleus gigas
Body parts measured : DPB , dorsal prosoma breadth ; DBL , dorsal body length ;
IOL , interocular length ; VPB , ventral prosoma breadth ; TL , telson length ; FMH , frontal
marginal height ; CH , central height ; FL , sub-frontal length ; Sl , length between dorsal
opisthosomatic spine and median mating scar ( Mts ) ; Gnb , gnathobases ; Ovd , oviducts ;
Gop , female gonopore ; Gol , genital operculum ; EM , egg-mass ; Anl , anal slit ; Doc , dorsal
ocelli; Lcy, lateral comp.eye;Chl, chelicera; Chi, chilarium,VBL, ventral body length.
 44

COMMON CURVE COMMON CURVE

0.45 for T. gigas for C. rotundicauda

0.40

0.35

0.30

0.25

O O
0Œ
Weight ( gm. )

0.20 + Π+
0
0.15

0.10
Log

0.50

O 0.5 1.0 1.5 2.0 0 0.5 1.0 1.5 2.0

Log Length ( cm. )

Fig .7. Parabolic growth curves of Tac

Fig.7 hypleus gig
ach as (A) and
gigas
Carcinoscorpius rotundicauda (B)indicating faster growth
rate in females than that of males.
 123
123 123
123 123
123 123
123 123
123123
123 123
123
123123 123
123123
123
123
123
123
123123 123
220 123 123123
123
123
123 550
123
123 123
1234
1234
1234 123
123123
123 123
123
123 1234
123 RANGE 1234 123123 123

T
123
123 1234
123 1231234 123123 123
123
123 123123
1234 1231234 123123 123
123
123
123
123
123
123123
1234
123123
1234
123
1234
SD 1231234
1231234 123123
123123
123
123
123
123 123
1234
123 1231234
1234 123123
123 123
123
123
123 123
1234
123 1231234 123123 123

T
123
123 1234
123123 1231234 123123 123

T
123
123 123123
1234 MEAN 1231234 123123 123

T
123
123 1234
123123 1231234 123 123
123
123 123123
1234 1231234 123 123
200 123
123 123123
1234 1231234 123 123 500
123 123123
1234 1231234 123 123
123 1234 1234 123
123
123 123
1234
1234
123 1231234
1231234
123
123
1234 123
1234 123

O
1234 123

T
1234 123
♀
1234 1234
1234
123
123
1234
1234 123
123
1234
1234 123
123
123 1234 123 123
123 1234 123 123
180 123
123
1234
1234
123
123 123
123
450
123 1234 123 123

T
123 1234 123 123
123
123 1234
1234 123 123
123
123 1234 123 123
123 1234 123
123 123
123 1234 123
123 1234 123 123
123
123
1234 123 123
123
123 123 123

T
123

T
123 123 123
123
123 123 123 123
123
123 123 123 123 400
160 123 123 123 123 123
123
123 123
123 123
123
123
123
123
123
123 123 123 123 123
123
123 123 123 123
123
123 123 123 123
123 123

T
123 123
123 123 123 123
123 123
123 123
123
123
123
123 123 123 123

T
123 123 123
123
123 123 123
140 123
123 123 123
123 123
123 350
123 123
123
123 123
123
123
123
T
123
123
123
123
123
123
123
120 123
123 300
123
123
123
123
123
123
123 123
123 123
123 123 123
123
123 123 123
123
123 123 123
123
123
123
123 123
123
100 123 123 250

T
123
123 123
123
123
123 123
123
123
123 123
123
123
123
123
123 123
123
T

123
123 123
123 123
123 123
123
123123
80 123
123123 123
123 200
123
123123 123 123
123123 123 123
123123 123 123
123123
123 123
123 123
123
123 123 T 123
123 123 123
T

123 123 123

123 123 123

T
123
123 123
123
123
123 123
123 123
123
60 123
123 123 150
123 123 123
123
123 123
123 123
123
123 123
123
123
123
123
123 123
123 123 123
123
123 123
123
T

123 123
T

123 123
123 123
123 123
40 123 123
1234 100
123 1234
123 1234
123 1234
123 1234
T

123 1234
T

123 1234
123
123 1234
1234
123
123
123
20 123 50
123
123
T

123 Ä
Ä 123
123123
123123
123123
T

123
in gm
FMH

DBL

VPB

DPB

IOL

Wt.
CH
RL

FL

TL

Different Morphometric Variables

Fig.8. Graphical presentation of Nine (9) morphometric variables ( mm ) and

body weight (gm) of 23 mated males and females of Tac achhypleus
gigas
gigas..
 46

Mating Scars

A B

Trailing Scars

D
C
 47

Fig. 16 . (A-D) (p.46)

A , A male C. rotundiacauda in ventral view showing its prosomatic appendages , comparatively

broad opisthosoma with very small marginal spines . The telson is comparatively larger and stout ; ; B
Dorsal view of a gravid female, 40cm. in length including the spine ( T. gigas), showing three vivid
corroded marks , the ' mating scars ', C , Dorsal view of a juvenile horseshoe crab of approx. 5 cm.
length ,whose sex was not idtifiable; D, Ventral view of a female C. rotundicauda ( 16 cm. excluding
the spine) showing its prosomatic as well as opisthosomatic appendages ( genital operculum and gill
flaps );irregularities in the trailing edges of the opisthosoma due to damage to the carapace during
amplexus i.e. most of the movable spines were dropped off.

A B

Fig.17.

A , One male T. gigas is in upturned posture ;

B , Adult horseshoe crabs ( T. gigas ) rest on
the sand beach of CHANDIPUR, ORISSA for
taking individual weight with a pan-balance.
 48

Male Female

A.
Ventral view of
Carinoscorpius rotundicauda
G

A * H B
Carinoscorpius rotundicauda

s
ite
s
dite
po
d po
ro
ro p
Tachypleus gigas

p ale
ale 1st pair fem
fem
I J K L

rs
mal

2nd pair
spe
cla
e cla

ale
sper

m
E
s

C D
F

B. Morphological Differences & Sexual Dimorphism in C. rotundicauda and T. gigas.

 1st Walking leg 1st Clasper organ

2nd Clasper organ

2nd Walking leg

Female Male
Genital operculum

B. Ventral Appendages of Tachypleus gigas

1st Clasper organ 1st Walking leg

2nd Clasper organ 2nd Walking leg

Male Female
D. Ventral Appendages of Carcinoscorpius rotundicauda

Fig. 18 (A-D)
A, Ventral view of C. rotundicauda showing comparative body size
of the male (smaller ,left one) and the female(larger , right one); B
& D, Comparison between five walking legs : 5B, with genital
operculum in T. gigas and 5D , first walking legs with attached
chelicerae in C. rotundicauda; C, Shows the morphological difference
and sexual dimorphism in two species of horseshoe crabs . A & B ,
dorsal carapace ; C & D , structure of genital operculae ; E & F ,
telson structures ; (*H) in T. gigas only ; and I,J & K,L show the
most distinguished characters of sexual dimorphism .
 50
DISCUSSION

The publication of vander Hoeven (1838) provided the first comprehensive treatise on the
morphology of horseshoe crab " Limulus moluccanus " , presently known as T. gigas and thereafter
only few fragmentary informations on morphology , including that of the larvae , were added by Packard ,
Lankester , Patten and others in the latter half of the 19th century . From 1927 to 1932 , Shoji studied the
American and Japanese horseshoe crabs from the view point of comparative morphology and comparison
of their larval forms just after hatching , and in 1929 he gave an account of the anatomy of Tachypleus
tridentatus in detail . The morphology of Limulus polyphemus in the modern sense was given by
Snodgrass in 1952 .
Recently a vast collection of information regarding horseshoe crabs morphology con-
sists of data on intraspecific variation , specially in body size and length of marginal spines . The morpho-
logical variation was given in L. polyphemus by Shuster (1957) , in T. gigas by Sekiguchi et . al . ,
(1976) , and C. rotundicauda by Sekiguchi et. al. (1978) .
The body of the horseshoe crab in distinctly divided into two parts , an anterior large
prosoma or cephalothorax and a posterior , small and hexagonal opisthosoma ; the latter is freely movable
up and down with its broad base attached to the prosoma on a strong transverse hinge . The dorsal surface
of the body is elevated along the middle . The glossy carapace is hard and is composed of an organic
combination of chitin and protein (Lafon , 1941) . The telson , sharply elongated posteriorly , together
with marginal spines arranged along the postero-lateral margins of the opisthosoma , gives the animal a
unique appearance . In outline , the prosomatic carapace is suggestive of a horseshoe as used in the
common name of the animal . The dorsal convexity of the prosomatic carapace enables the animal effec-
tive for feeding and respiration when the animal is embedded in muddy or sandy substratum . The swelling
of the carapace is moderate in T. gigas and weak in C. rotundicauda . The antero-frontal margin forming
an arc of the prosoma is relatively more in female than in the males . But in C. rotundicauda this arc is
almost same in both sexes . Only the male of T. tridentatus has a pair of notches on this arc antero-
laterally (Sekiguchi , 1988) .
The dorsal ocelli or dorsal eyes , sometimes called as median eyes , are a pair of very
small sports situated at the starting point of the median ridge . The distance between the two ocelli varies
according to the species ; they are far apart in L . polyphemus , moderately distant in T. gigas and C.
rotundicauda , and close together in T. tridentatus (Yamasaki et al. , 1988) .Compound , lateral eyes are
situated on and near the opthalmic ridges . The average diameter of a compound eye is more than 10 mm.
in L. polyphemus , whereas it is below 10 mm. in Asiatic species . Yamasaki , et. al . , (1988) showed the
progression in three Asian species as C . rotundicauda , T . gigas and T. tridentatus . It can be said
that C. rotundicauda has large eyes relative to its small body size . According to Waterman (1954) , the
maximum number of ommatidia was 1,161 in L. polyphemus and 446 in T. tridentatus. In C.
rotundicauda the distance between the two compound eyes (IOL) is shorter than in T. gigas .
Ratios of male to female morphometric characters (Table -4) indicate that it is 0.7787 +
0.0373 and 0.8460 + 0.0373 in T. gigas and C. rotundicauda, respectively . These results actually
tell the comparative body size of male and female that is in the former species the male is nearly 78% of
the female , while in the latter species it is nearly 85% . But in respect to body weight the male T. gigas
is almost 37% of the female , while it is 52% in C. rotundicauda . In every aspect it is evident that the
female specimens of C. rotundicauda are smaller than the females of T. gigas , but the situation is
reversed in case of male . Perhaps the comparatively large body of female has potentially effective for
lengthening the breeding cycle in the estuarine environment . This has also been further confirmed by the
availability of C. rotundicauda in Sundarbans right from Frbruary to August or Spetember , at least
solitary females on the muddy shore .
 51

Among the four morphometric variables , e.g. interocular length (IOL) dorsal prosoma (DPB) ,
dorsal body body length (DBL) and ventral prosoma breadth (VPB) , simply or partially correlated
(Table - 5) , it was the DBL and DPB , between which the perfect correlation had occurred . In the 2nd
r
step , it is only 23.14 where both sex category of either species showed significant P (0.05 , P < 0.01) .
It is , therefore , justifies to make a length-weight relationship (Le Cren , 1951) considering only DBL as
size parameter with that of body weight .

In the present discussion horseshoe crab , T. gigas showed n = 3.08995 and 2.52090 for
males and females respectively ; while it was 3.09554 and 2.25312 in C. rotundicauda . They may be
vital units of taxonomic consideration of the species . Presently it has been found that T. gigas , the
males increased in weight by an exponent of 3.08 and females by 2.52 ; while in C. rotundicauda males
and females increased by an exponent of 3.09 and 2.35 ; respectively . Although these 'n' value higher in
males than in females but did not differ significantly (Table -12, 't' test) . Such higher values for males
were common in fishes too (Sekharan , 1968 ; Chanchal et . al . , 1978 ; Lazarus and Reddy , 1986).

A female of T. gigas A female of C. rotundicauda

In every aspect it is evident that the female specimens of C. rotundicauda are smaller
than the females of T. gigas , but the situation is reversed in case of male . Perhaps the
comparatively large body of female has potentially effective for lengthening the breed-
ing cycle in the estuarine environment . This has also been further confirmed by the
availability of C. rotundicauda in Sundarbans right from Frbruary to August or
Spetember , at least solitary females on the muddy shore .
 52

An excellent review of length-weight relation was presented by Peters (1983) where Pough (1980) showed
the ranges of regression coefficient were 2.98 - 3.02 in reptiles , 2.94 - 3.64 in salamanders , 2.69 in
turtles , 3.24 in frogs ; while Jerrison (1973) showed 2.53 in world record of fishes and Smock (1980)
recorded 2.62 in aquatic insects . McMahon's (1973 , 1980) explanation of the basal metabolic rate (Rb)
in terms of body mass (Rb α W 3/4) has been deduced from the length-weight relation and he finally
concluded that Rb varied as body mass to the three-fourth power (i.e. W3/4). So in this context the present
investigation may open a new line to manage horseshoe crabs in captivity .

Deducing the two parabolic growth equations of T. gigas and C. rotundicauda it is found
that the former species will stop its growth completely when it attains 248 mm in maximum length and
766 gm in maximum weight , while the latter species ceases growth at 247.28 mm and 756.40 gm of
maximum length and weight , respectively . The above assumptions are simply the theoretical values
which usually do not corroborate with the natural findings ; as for example , horseshoe crabs in Indian
coastal region observed maximum 629 gm in weight and 209.6 mm in DBL . Increase in weight was found
to follow the cube law in females of C. rotundicauda and two curves of males and females intersected
between the carapace length 130 - 135 mm which indicated the length in which the animal attains sexual
maturity (Chatterjee et. al. , 1988) . But in the present study it has been found that the females reach
maturity sooner than the males , and growth of the female is even faster in C. rotundicauda ; while
combined growth patterns are more or less identical in both species (Fig. 7) .

Biometric analysis of Tachypleus (T.gigas) matting indicates the following points :

1) Large males tended to 'prefer' large females and medium-sized males 'prefer' medium fe-
males . Small males , however , donot show a "preference" for small females , but rather for medium-
sized ones ( Table - 8 A,B,C) Although X2 -test couldn't be done due to the appearance of expected value
more frequently less than 5 (Bailey , 1959) . Su\huster , el. al. , (1992) has confirmed this hypothesis in
L. polyphemus , where in no case the male size was homogamous with that of the female partner
(Pomerat , 1933) ; Cochen & Brockman (1983) and Brockman (1990) also demonstrated assortive
matting in L. polyphemus . According to them (opt. cit.) competition between males for females was also
independent of size i.e. , attached males were not significantly larger than the unattached males .

ii) It has already been established that the female horseshoe crab is larger than the male
(shuster , 1982 ; Sekiguchi 1988) , it might , therefore , be deduced that higher correlations represented
closer approximations in size for characteristics studied (Shuster et. al. 1992) . If this is true , then , if
a charteristic of one member of a mated pair diverges from the overall male to female regression line ,
then it is specific with sex . Hence , sexual dimorphism occured in the above morphometric features . But
less significant levels and low coefficients (Table -7 ) make such conclusions dubious .
iii) There are three " mating scars " an axial one with two laterrals on either side (Fig. 6)
are caused by the motion of the male prosomal arch abetted by sand , while amplexus has been in a long
time (Shuster , 1955/58) . These " mating scars " are mute evidence of the duration of amplexus and
the numbers , and positions of scars can be used to idicate the age of the female as well as the size and
the number of previously attached suitors (Shuster et. al. , 1992) .
 53

In the present study it was also ecident that in terms of weight the male assumed 35% of
the total body weight of the female , while in all other morphotic characteristics the males were of 75.8%
of the females ( Table -16) . The total egg mass of gravid females varies from 55 to 195 gms with a mean
value of 133.56 gm. It was found after serveral trials that 1 gm of eggs counted 28 in number which will
help to determine the total egg-number in a female . For instance , a female T. gigas of 505 gm total body
weight yielded 5,497 eggs , while another female of same weight gave the figure as 3295 . Interestingly
enough , a female severely infested with epifaunal associates (confirming its old age) with only 355 gm of
total body weight gave only 9 mature eggs (brownish , rather than usual light yellow-greenish
appearence) . The wide variation in egg numbers contained by the females is indicative to the frequent and
usual egg laying behaviour of the animal. When regression as well as correlation were made between
DPB of male and female egg mass it showed a very weak but negative coefficient ( r = -0.224 )
(Table -17) . From the same table -17 it was found that only CH , FMH and RL show significant
correlation and comparatively higher tangent angles of regression lines .

Comment :
Since all the four living adult horseshoe crabs are not
known to molt, the term " growth " in adults is an area of
enquiry needing more study.

......... Prof. C.N. Schuster, Jr. (1993)

An excellent review of length-weight relation was presented by Peters

(1983) where Pough (1980) showed the ranges of regression coefficient were
2.98 - 3.02 in reptiles , 2.94 - 3.64 in salamanders , 2.69 in turtles , 3.24 in
frogs ; while Jerrison (1973) showed 2.53 in world record of fishes and Smock
(1980) recorded 2.62 in aquatic insects . McMahon's ( 1973 ,1980) explanation
of the basal metabolic rate (Rb ) in terms of body mass (Rbα W 3/4) has
been deduced from the length-weight relation and he finally concluded that Rb
varied as body mass to the three-fourth power (i.e. W3/4). So in this context
the present investigation may open a new line to manage horseshoe
crabs in captivity .
 54

C . Absolute population estimates

 55

INTRODUCTION

It is usually possible to estimate the size of population and communities of sessile and
sedentary organisms by using methods which involve fixed sampling units i.e. , quadrate or line-transect
methods . But with mobile individuals i.e. , insects , fishes , snails , horseshoe crabs and other animals
different procedures , particularly capture-recapture techniques are required .
The application of recapture method for such purpose was first made by Lincoln (1930)
following the principles of proportion of capture-recapture of marked animals introduced by Peterson as
early as 1889 . Lincoln estimated the total number of ducks in North America , and the formula he used
was sometimes called the "Lincoln Index" . The principle is very simple : a number of animals are
captured , marked and released . When the marked animals are thought to have mingled thoroughly with
the unmarked population , a further sample is captured and the proportion of marked individuals noted .
Then an estimate of the total population size is given by duviding the number of marked animals released
by the proportion found to be marked in the second group of captures . Jackson (1937 a,b) independently
used the same method for estimating the density of tsetse flies . In a subsequent extension of this method
Jackson (1937 , 1939) used his "positive" and "negative" methods allowing certain range of complications
imposed due to birth-and death-rates . There are also certain other methods of estimating a population that
depened on the presence of marked individuals , employing different principles to the Lincoln Index . A
comprehensive survey of these marking techniques as well as capture-recapture methods has been re-
viewed by Seber (1973) .There are at least eight assumptions (Southwood , 1978) which underline all
methods of capture-recapture analysis , such as :-

1. The marked animals are not affected (neither in behaviour nor in life expectancy) by being marked
and the marks will not be lost .

2. The marked animals become completely mixed in the population .

3. The probability of capturing a marked animal is the same as that of capturing any member of the
population sampled randomly with respect to its marked status , age and sex , termed as 'equal catchability'.
This assumption has two aspects : i) that all individuals of the differen age groups and of both sexes are
sampled in the proportion in which they occur , ii) that all the individuals areequally available for capture
irrespective of their position in the habitat .

4. Sampling must be at discrete time intervals and the actual time involved in taking the samples must be
small in relation to the total time .
5. If the population is not a closed one , immigration and emigration can be measured or calculated .
6. There are no births or deaths in the period between sampling or , if there are , allowances must be
made for them . A few horseshoe crabs unlike Limulus become stranded .So the number of death is
is negligible in this study.
7. Being captured one or more times does not affect an animals subsequent chance of capture. This
is a further extension of assumption no.3 i.e. , equal catchability .
8. Every marked animal has the same probability of surviving through the sampling period .

Comment : Unless animals of the same age are

marked , survival probability becomes
biased.............. Prof. C.N.Shuster, Jr.( 1993 ).
 56

Fig. 19 (A-C)
A,Shows the making device with steel needle on the dorsolateral
carapace of one horseshoe crab (T. gigas); B, Shows the coded
mechanical mark on the right dorsolateral surface of prosoma;
C , One marked and segregated female of T. gigas released in the
falling tide .
 57

OBSERVATIONS

A. Application of Jackson's Principle in the present investigation :

Horseshoe crabs are mobile in nature and they prefer a wide territorial range (habitat) in
the sea particularly in the shelf region , come to the shore only during the breeding season , mostly in
pairs in Indian coastal region (Debnath , 1985) . So their population size was measured only by capture
- recapture method and it was not possible to estimate the population size of such mobile animals by
quadrate method or by using any other fixed sampling units . This author used Jackson's "Negative" and
"Positive" method (Jackson , 1939) to estimate the population size of adult horseshoe crabs , Tachypleus
gigas that came to breed at Chandipur sea shore in Orissa State from March to July , 1985 . The
principle of "Positive" method is , in contrast to the "negative" method , to release a large number of
marked animals on a single occasion . Capture is then made on a number of later occasions , the
number of marked animals being recorded each time . The majority of animals captured on these later
occasions should be released again , otherwise one has to consider special account to the diminution in
the number of marked animals available for recapture . On the other hand , the principle of the negative
method is to catch , mark and release groups of animals on several successive observation days , making
no record of the numbers of recaptures until the last occasion . Such a method is suitable when the
preliminary marking of successive batches can be carried out by relatively unskilled workers (Bailey ,
1952) .
However , the idea of Jackson's captur-recapture method is simple enough , but it is very
difficult to attribute a precise variance to the estimates (Bailey , 1952 ; Andrewartha , 1970) .

CALCULATIONS AND RESULTS

a) Crude - Recaptures :

The complete data showing the numbers of captures , releases and recaptures can be
demostrated by a triangular array of " trellis" diagram . Three "trellis" diagrams have been constructed
to accommodate the crude-recaptures ( Table 13 A-C) in which total of 653 , 1082 and 680 individuals of
horseshoe crabs were counted in March , May and July , respectively . Although 692 , 1120 and 703
animals were captured , 39 , 38 and 23 had been rejected because of their moribund condition or
physical injury caused by the ravens Corvus splendens , an effective predator bird (Debnath and Choudhury
, 1988a). The numbers in each row of the Table 13 A-C (horizontal) indicate the number of horseshoe
crabs marked on the date indicated in the left margin . The vertical columns indicate the numbers recap-
tured on those that were marked on the dates shown in the left margin . In respective three months the total
number of animals recaptured were 73,307 and 107 . Thus the ratios of total captured , total marked and
released and total recaptured are 1.05:1:0.11 , 1.03:1:0.28 and 1.03:1:0.15 for the successive three months
as mentioned above .

b) Corrected Recaptures :

In Table - 14 A-C each row provides the row material for estimating the size of the population
(on the day of release) by Jackson's "negative" method . The values in the body of the Table 14 A-C are
corrected recaptures which have been calculated with the help of the equation No. 1.
 59

Population size :

The values for 'r' calculated from each row and column by using equation 2 or 3 (equation 2 that
covers 3 entries only). Equation No. 4 is then used to calculate 'a' for each row and column and the
population size N obtained by using equation No. 5 . Thus the table shows the calculated values for
r+ , r , a0 and N. In March 1985 , the number of individuals that arrived on the 7th , 8th and 9th date
-
respectively were calculated by positive method , while for the 10th date it was calculated by negative
method . Similar methods were followed to estimate the population of T. gigas in May and July
(Table 15) .
 59

Equations:
100 100
Yn = Rn X X ......................( 1 )
Cm Cn

Y2 + Y3 + Y4 +......+ Yn
r+/- = ......................( 2 )
Y1 + Y2 + Y3 +......+ Yn-1

Y3 + Y4 + Y5 +........+ Yn
r+/- = ......................( 3 )
Y1 + Y2 + Y3 +........+ Yn-2

( Y1 + Y2 + Y3 +........+ Yn-1)
a 0 =
r+/-
- ( Y1 + Y2 + Y3 +.......+ Yn-2) ........( 4 )

10, 000
N+/ - = ......................( 5 )
a 0

Where , Yn = Corrected recaptures ,

Rn = No. of recaptures on date n ,
Cm = No. of marked on date n ,
Cn = Total captured (including recaptures) on date n ,

Yn
r = The weighed ratio of Yn-1

a0 = Corrected values for the date of estimation ,

N+/- = Estimated population on date n.

Animal dispersion and sexual selection :

Out of the total 50 amplexed animals released within 50-200 mts. , 9 pairs of T. gigas were
recovered from Burhabalanga sand flat , 8 at a distance of 200 mts or within from the point of release
and other pair at a distance of 1000 mts opposite to the site of release . These findings (Table 22) ,
recovery rate , R = 18 % ) helped to document the dispersal behaviour of the species , however , in the
restricted sea shore only . One interesting observation was made that out of 15 mating pairs segregated
and released , one amplexed pair was recovered after 3 days from the date of release and they showed
similar marks on their cararapaces (Debnath and Choudhury , 1988a) .
 60

B. Application of Lincoln 's method :

In the present part of the tagging study the horseshoe crabs were captured , marked and
released at a time and then recaptured in the following day only . Between the receding tide of 26th
March and the following high tide of 27th March provided almost 24 hrs. to intermingle the released
population with the unmarked population that required to be determined . The calculation is very simple
enough following the Lincoln - Peterson Index as shown below :

a.n
<

N = Where , a = total marked and released .

r
n= total second sample including recaptures (r)
<

Thus , N = 300 X 284 = 4,260

20
Bailey (1951 , 1952) has suggested that with small samples (if r < 20) a less biased
estimate can be obtained if 1 is added to n and r , thus ;

a (n+1)
<

300 (284 +1)

N = = = 4071.42
r +1 20 + 1
The 'variance' and the 'standard error' of N are :

a2 (n +1). (n - r )
<

Var N = = 697959.18
( r + 1)2 . (r + 2)
<

S .E . = Var N = + 835.439

( C ) Application of Jolly - Seber stochastic method :

Jolly (1965) and Seber (1965) have , independently , developed a common method to
cover situations in which there is both loss (death and emigration) and dilution (birth and immigration).
Their methods give similar solutions , except that Jolly's makes allowance for any animals
killed after capture and hence not released again . At Digha (West Bangal) horseshoe crabs population
comprising only adult T. gigas has been severely affected by human interferrence and a predator , C .
splendens ( Debnath, 1985 ; Debnath and Choudhury , 1988a) . So the factor , like "loss" due to death
and emigration is more inevitable than "dilution" . However , dilution in the population may occur to some
extent due only to immigration but not with birth . The Jolly-Seber method efficiently groups the data and
is fully stochastic , " an extra advantage over any deterministic method , however its reliability strictly
depends , effectively , on the probability of any animal surviving through any period is not affected by
its age at the start of the period". The Jolly - Seber 's estimate of population is usually reliable when
9% or more of the population is sampled and the survival rate is not less than 0.5 (Bishop and Sheppard ,
1973) .
Although this method may seriously overestimate the survival rate , it properly remains the
most useful method to date and has been used for a number of different insect population (Parr , 1965 ;
Sheppard et. al . , 1969 ; Fletcher , 1973 ; Ito et. al. , 1974)
 61

Calculation and Results :

The basic equation in Jolly's method (1965) is :

^ ^ .n
M
Ni = i i ...................................... (I)
ri

^ ^
where , Ni = the estimate of population on day i , Mi = the estimate of the total number of marked
animals in the population on day i (i.e. , the counterpart of 'a' in the simple Lincoln Index) , ri = the total
number of marked animals recaptured on day i and ni = the total number of animals captured on day i .
The procedure of calculation may be demonstrated as follows :

1) The field data are tabulated as in Table 16 A according to the date of initial capture or mark
and the date on which the animals were last captured . The columns are then summed to
give the total number of animals on the ith occasion ( = Si of Jolly , 1965) , recaptured
subsequently (Ri)

2) Another table is drawn up (Table 16 B ) giving the total number of animals recaptured on day i
bearing marks of day j (bold printed in the Table 16 B) or earlier (Jolly's a i j ); this has been done
by adding each row in Table 16 B from left to right and entering the accumulated totals . The number
marked before time i which are not caught in the ith sample , but are caught subsequently (Zi) , is found by
adding all but the top entry in each column . The top entry in each column represents the number of
recaptures (ri) for the day on its right .

3) Then the estimate of the total number of marked animals at risk in the population on the
samplind day is obtained thus (Table - 17)
ai . Zi
^ i ......................... (II )
Mi = +r
Ri

4) The proportion of marked animals in the population at the moment of capture on day "i" is
found and enterd in the final table (Table 17) , thus :
ri
∝i = ..................................................(III)
ni
5) The total population is then estimated for each day by the following equation as constructed
using equation -I and II :
^
M
^
N = i .................................................. (IV)
i ∝i
 62

6) The probability that an animal is alive at the moment of release of the ith sample
will survive till the time of capture of the sample ( i + 1) th sample is obtained :

^
M
^ i+1 ...................... (V)
Øi =
^ -r +a
M i i i
For technical as well as biological reasons this survival rate may be converted into a "loss
rate" ( the effect of the predation , Debnath and Choudhury , 1988a ) and emigration due to
human interference .

^ ^
γ = 1 - Ø i ..................................( VI )
i

7) The number of new animals joining the population ( due to fresh emergence of T. gigas
from the shallow shelf region and from adjoining areas) in the interval between the ith
and (i+1)th samples and alive at the time (i+1) is given by :

^ ^ - ^( ^
B i = N i+ 1 Øi N i - n i + a i) .......( VII )
^
This equation no. VII may be converted into the dilution rate, â , thus :

1 ^
B
i ................................( VIII )
= 1 -
^
â ^
N i+ 1

8) Now in the final calculation (Table 17) standard errors, i.e. the square root of the variances
of N^i, Ø ^ , and B^ are obtained by using very cplicate equations ( discussed in detail by
i i
Jolly, 1965 ).

^ ) and survival rate (var^ )

9) According to Jolly(1965) the variances of the population(var N i Øi
estimates given above contain an error component due to the real variation in population
numbers. These errors of estimation may be obtained with the following formulae:

^
M i- ri + a i 1 - αi
i) var ( ^
Ni / N i ) = ^
Ni ( ^
Ni - ni ) - x ( 1 - 1 ) +
M Ri ai ri
i
 63

^2( 1 - ^ )
^/ ) = ^) - Øi Øi
ii ) var ( Ø
i Øi
var ( Ø
i ^
M i+1

Now , results obtained by mathematical equations following procedures , number 3 to 9 are tabulated in
the final Table -17 . The special problems involved in the computation of the summation term of var ( N )

What the author contends is that an organ-

ised team with working facility in the feild and
assistance for a longer duration taking many
more stations along the coast may help to bring
out a more comprehensive population picture
of the Indian horseshoe crabs .

But an interesting observation in recovering

two segregated opposite sexes further
amplexed (coupled) 3 days after release has
rendered the author to assume that specific
sexual selection might had occurred during
the breeding activities. But how they choose
themselves and found together and returned
to the breeding ground remained a great
mystery .
 64

Table 13 (A, B & C) .

Showing crude recaptures in March and July , 1985 at Chandipur (Orissa)

Trellis Diagram . 13 A
Date Total Total DATE RECAPTURED Total
marked captured marked & recaptured
released 7th 8th 9th 10th 11th 12th 13th

March,6th 12 11 1 2 - - - - -
7th 70 68 4 18 2 - - -
8th 193 188 26 12 - 1 -
9th 202 194 2 2 1 -
10th 74 72 1 - -
11th 63 60 - 1
12th 50 36 -
13th 28 24 1 6 44 16 3 2 1 = 73

No. captured 692 653 68 188 194 72 60 36 24 = 642 + 11

Date captured. March'1985 7th 8th 9th 10th 11th 12th 13th = 653

Ratio of total captured , total marked & released and total recaptured , respectively = 1.05 : 1 : 0.11
 65

Trellis Diagram 13 B
Date Total Total DATE RECAPTURED Total

marked captured marked recaptured

& released

May' 1985 2nd 3rd 4th 5th 6th 7th 8th 9th 10th 11th 12th

1st 5 4 - 1 - - - - - - - - -
2nd 12 12 2 1 - 1 - - - - - -
3rd 83 80 28 15 - 1 - - - - -
4th 186 184 32 27 12 8 - - - -
5th 197 190 32 24 7 - - - 4
6th 223 218 24 19 8 - - 5
7th 155 152 17 7 13 - -
8th 92 88 8 5 7 -
9th 57 54 2 1 -
10th 51 48 2 -
11th 36 34 -
12th 23 18 0 3 29 47 60 64 51 23 20 10 9 = 307

No.Captured 1120 1082 12 80 184 190 218 152 88 54 48 34 18 = 1078+4

= 1082
Date Captured
May'1985 2nd 3rd 4th 5th 6th 7th 8th 9th 10th 11th 12th

Ratio of total captured , total marked and released and total recaptured respectively = 1.03 : 1 : 0.28

Trellis Diagram 13 C
Date Total Total DATE RECAPTURED Total
marked captured marked & released recaptured

June - July'1985
30th 1st 2nd 3rd 4th 5th 6th

29th 9 8 1 1 - - - - -
June - July'1985

30th 59 56 3 20 4 - - -
1st 97 94 18 16 3 - -
2nd 185 182 15 10 2 -
3rd 167 164 4 5 1
4th 98 94 1 -
5th 63 60 1
6th 25 22 1 4 38 35 17 8 2 = 105
No. captured 703 680 56 94 182 164 94 60 22 = 672 + 8
Date captured
June-July '1985 30th 1st 2nd 3rd 4th 5th 6th = 680

Ratio of total captured, total marked and released and total recaptured respectively = 1.03 : 1 : 0.15
 66
Table 14.
Corrected recaptures in March and July,1985 at Chandipur,Orissa.

A. (7 th - 13 th March'1985)

Date marked DATE RECAPTURED N

& released(DMR) 7th 8th 9th 10th 11th 12th 13th (+)
6th 13.368 9.671 - - - - - -
7th 3.128 13.644 4.084 - - - 785
8th 7.128 8.865 - 1.477 - 1,144
9th 1.431 1.718 1.431 - 5,820
10th 2.314 - - -
11th 6.994 -
12th - -
13th
N (-) 820 Average = 2142
Date captured 7th 8th 9th 10th 11th 12th 13th

B. (2nd - 12th May'1985)

(DMR) 2nd 3rd 4th 5th 6th 7th 8th 9th 10th 11th 12th N(+)
1st - 31.251 - - - - - - - - - -
2nd 20.832 4.524 - 3.816 - - - - - - 178
3rd 19.005 9.862 - 3.285 - - - - - 225
4th 9.139 6.714 4.281 4.934 - - - - 1,042
5th 7.709 8.293 4.182 - - - 11.687 3,850
6th 7.222 9.885 6.782 - - 12.721 1,857
7th 12.687 8.512 17.790 - - 2,273
8th 16.821 11.831 23.386 - 1,541
9th 7.710 5.443 - -
10th 6.126 - -
11th - -
12th -
N(-) - - - - 2583 1946 2284 4076 368 206 - 1725

Date captured 2nd 3rd 4th 5th 6th 7th 8th 9th 10th 11th 12th

C. (29th June - 6th July)

(DMR) 30/6 1st 2nd 3rd 4th 5th 6th N(+)

29/6 22.312 13.287 - - - - - -

30/6 5.692 19.599 4.348 - - - 476
1st 10.504 10.357 3.389 - - 473
2nd 5.015 5.835 2.029 - 980
3rd 2.589 5.070 2.767 2042
4th 1.771 - -
5th 7.571 -
6th
N (-) - - - 935 1490 1873 - 1181

Date captured 30/6 1st 2nd 3rd 4th 5th 6th

 67

Ta b le 15.
The values for r+ , r- , a0 and N in March , May & July 1985 at Chandipur
(Orissa)

POSITIVE METHOD NEGATIVE METHOD

r a0 N(+) r- a0 N(-)
+

1985,March 7th 1.0570 12.7395 785 Average of March= 2142

8th 0.6466 8.7387 1144
9th 1.0000 1.7180 5820
10th 0.7951 12.2018 820

1985,May 2nd 0.3281 56.2573 178 Average of May = 1725

3rd 0.4552 44.3782 225
4th 0.7919 9.6001 1042
5th 0.8861 2.5972 3850
6th 0.8163 5.3861 1857 1.3692 3.8712 2583
7th 1.2402 4.3991 2273 1.2481 5.1392 1946
8th 1.2291 6.4891 1541 1.4083 4.3783 2284
9th 1.6562 2.4531 4076
10th 0.6593 27.1582 368
11th 0.4012 48.4533 206

1985,July 30/6 0.9462 21.0187 476 Average of July = 1181

1st 0.6583 21.1552 473
2nd 0.7181 10.2043 980
3rd 1.0233 4.8962 2042 0.9773 10.6972 935
4th 0.9131 6.7103 1490
5th 0.9632 5.3378 1873
 Table 16.
A. The tabulation of recaptured data according to the data on which the
horse shoe crabs were last caught for analysis ( afterJolly , 1965)

Date of Total Total

capture capture released
i ni ai

d1 48 48 1
d2 57 56 2 2
d3 83 80 1 2 3
d4 187 185 0 1 28 4 Day when last
d5 197 192 0 0 15 32 5 captured (j)
d6 225 224 0 1 0 27 32 6
d7 260 255 0 0 4 12 24 22 7
d8 147 142 0 0 0 8 7 19 17 8
d9 92 90 0 0 0 0 0 8 9 8 9
d10 68 68 0 0 0 0 0 0 13 4 17 10
d11 107 100 0 0 0 0 1 6 2 0 2 9 11
d12 56 56 0 0 0 0 2 0 1 3 0 4 13 12
d13 38 36 0 0 0 0 3 5 2 0 4 2 4 11 13
d14 49 48 0 0 0 0 0 0 5 3 0 0 1 2 10 14
d15 57 56 0 0 0 0 0 7 0 1 5 0 4 0 5 8 15

Ri = 3 4 47 79 69 67 49 19 28 15 22 12 15 8
68

R2 R3 R4 R5 R6 R7 R8 R9 R10 R11 R12 R13 R14

 B. Calculated table of the total number of marked animals recaptured
on a given day bearing marks of day or earlier (after Jolly , 1965)

d1 1
d2 2 2
d3 1 3 3
d4 0 1 29 4
d5 0 0 15 47 5
d6 0 1 1 28 60 6
d7 0 0 4 16 40 62 7
d8 0 0 0 8 15 34 51 8
d9 0 0 0 0 0 8 17 25 9
d10 0 0 0 0 0 0 13 17 34 10
d11 0 0 0 0 1 7 9 9 11 20 11
d12 0 0 0 0 2 2 3 6 6 10 23 12
d13 0 0 0 0 3 8 10 10 14 16 20 31 13
d14 0 0 0 0 0 0 5 8 8 8 9 11 21 14
d15 0 0 0 0 0 7 7 8 13 13 17 17 22 30 15

Z (i-1) = 1 2 20 52 61 66 64 58 52 57 46 28 22 ri
Z2 Z3 Z4 Z5 Z6 Z7 Z8 Z9 Z10 Z11 Z12 Z13 Z14
69
 Table 17 .
Estimated population of T.gigas at Digha sea shore by Jolly - Seber (1965) method

Days of Proportion No. marked Total popu- Survival No. of new STANDARD ERRORS Standard errors due toerrors in the estimation
May , 1986 of recaptures animals at lation rate animals of parameter itself
risk

∝i Mi Ni 0i Bi V(Ni) V(0i) V(Bi) V(Ni / Ni) V(0i / 0i)

17 - 0 - 0.3333 - - 0.3213 - - 0.3139

18 0.035 16.0 457.142 0.0914 136.196 524.1970 0.0414 79.6189 523.7608 0.0231
19 0.036 6.404 177.888 0.9092 330.249 86.6849 0.0396 93.9395 85.6532 0.0240
20 0.155 75.835 489.258 0.8268 402.546 90.2383 0.0411 106.0502 87.4896 0.0327
21 0.238 191.695 805.411 0.7839 364.812 129.1030 0.0546 97.9702 125.9477 0.0497
22 0.266 263.940 992.255 0.9474 764.536 152.5308 0.0685 118.7688 149.2452 0.0676
*FM 23 0.238 405.469 1703.651 0.8844 27.525 303.0322 0.0878 290.7265 300.2092 0.0868
24 0.346 529.315 1520.812 0.3408 260.521 376.4252 0.0643 147.5546 374.4005 0.0614
25 0.271 211.428 780.177 0.9757 -219.801 183.5881 0.1274 154.2766 181.4503 0.1854
26 0.500 269.733 539.466 0.9188 1004.827 138.3489 0.1857 362.3066 136.3980 0.1850
27 0.186 279.091 1500.489 0.6618 - 408.718 426.4219 0.1098 249.4151 424.6594 0.5750
28 0.410 237.666 579.673 0.3628 - 89.815 176.0974 0.0882 39.2232 174.4444 0.0832
29 0.815 98.2 120.490 1.4825 171.814 204.1878 0.2562 78.8038 21.5786 0.2693
30 0.428 153.0 357.476 - - 129.9201 - - 128.5370 -
31 0.526 - - - - - - - - -

* FM , Full moon
70
 Table 18 .
Number of T. gigas in Eight Transects ( 102 mt) counted from
20th to 31st March , 1986 at Chandipur in area of 1 sq. Km.

Transect 20/3 21/3 22/3 23/3 24/3 25/3 26/3 27/3 28/3 29/3 30/3 31/3 Population in tod*
(10 X 10) ( 12 days )
sq. mt.

1 0 0 0 1 0 2 1 0 0 0 0 0 4 3333
2 0 0 1 0 0 1 1 0 1 0 0 0 4 3333
3 0 0 0 0 1 0 2 1 0 0 0 1 5 4167
4 0 0 0 1 0 0 0 2 1 0 1 0 5 4167
5 1 0 1 1 0 0 0 1 0 0 0 0 4 3333
6 0 0 0 0 0 0 0 2 0 0 0 1 3 2500
7 0 1 0 0 2 0 0 0 0 1 0 0 4 3333
8 1 2 0 0 0 0 0 0 0 0 1 0 4 3333
800 sq.mt. 2 3 2 3 3 3 4 6 2 1 2 2 Total.............. 3437

4432

2500

2500
3750
3750
3750
3750
5000
7500
2500
1250
2500
2500
--
--
--
--
--
--
--
--

1 sq.km.=106 sq.mt. Population in terms of area (toa* )

* toa , terms of area , and tod terms of days

71
 Table 19. Correlation and Regression between estimated populations (N) with
captured , marked and released (C) and recaptures (R) at Chandipur and Digha , India

JACKSON'S METHOD JOLLY - SEBER'S METHOD

Chandipur sea shore Digha sea shore
( March - July , 1985) ( May , 1986)

Correlations: N = 20 , r = 0.647 , df 19 , P< 0.01 N = 13 , r = 0.687 , df 12 , P < 0.01

Se = + 0.130, t = 3.600 , df 18 . P < 0.01 Se = + 0.146, t = 3.134 , df 11 , P = 0.01

Regression (I): y = 0.8822 X + 1.2854 y = 0.8578 X + 1.0709

tan 0 = 41.420 tan 0 = 40.620

Relationships between N and R

Correlations: N = 17 , r = 0.655 , df 16 , P< 0.01 N = 13 , r = 0.481 , df 12 , P > 0.05

Se = + 0.138 Se = + 0.213
t = 3.632 , df 15 , P < 0.01 t = 1.819 , df 11 , P > 0.05

Regression (II): y = 0.6608 X + 2.2259 y = 0.3621 X + 2.2871

0
tan 0 = 33.45 tan 0 = 19.900

Estimated y = 4.85 (Log) y = 3.26 (Log)

population (N) : 70,795 1,820

..........."unauthorised
"unauthorised sacrifice"
sacrifice"...............
because in India the "donor crabs" are not
returned to its natural habitat ; rather they
are sacrificed in whole for its total blood .
Like U. S - F. D. A. regulation , neither such
exists in India nor it's taken up in action
...........this is why the author has stressed
upon the term "unauthorised sacrifice.
 73

Table 20. Showing the relation between total marked and total recaptured
horseshoe crabs , T. gigas in respect to its sex with recovery rate .

MONTH Total captu- Sexual isolation Total re-cap- Sexual isolation and Correlation between
of 1985 red , marked tured and re- recovery rate (%) captured and recaptured
and released covery rate (%) animals , r*

MARCH 653 333 320 73(11.17) 38(11.41) 33(10.93 ) r = 0.699 , p < 0.05

MAY 1120 568 552 307(27.41) 165(29.04) 142(25.72) r = 0.739 , p < 0.01

JULY 680 350 330 105(15.44) 60(17.14) 45(13.63) r = 0.937 , p < 0.01

* "r" represents the correlation coeficient of the relationship

of day to day captured and recaptured animals

Amplexus occurs in the off-shore

shallow water but not in the mud
or sand-beach . This inference can
be made from the recovery of males
and females in isolation bearing
similar marks / rubber bands
days after capture , tagging and
segregation .
 74
FULL MOON DAY

110
1234567
1234567 FULL MOON NIGHT
1234567
1234567
1234567
123456
123456
1234567
1234567
123456 1234567
100 123456
123456 1234567
1234567
123456 1234567
123456
123456 1234567
1234567
123456 1234567
123456
123456 1234567
1234567
90 123456
123456 1234567
1234567
123456
123456 1234567
1234567
123456
123456 1234567
1234567
123456
123456 1234567
1234567
80
123456
123456 1234567
1234567
123456 1234567
Individual No. Tachypleus gigas

123456
123456 1234567
1234567
123456
123456 1234567
1234567
123456
123456 1234567
1234567
70 123456
123456 1234567
1234567 123456
123456 1234567 123456
123456
123456 1234567
1234567 123456
123456
123456 1234567 123456
123456
123456 1234567
1234567 123456
123456
60 123456 1234567 123456
123456
123456 1234567
1234567 123456
123456
123456 1234567 123456
123456
123456 1234567
1234567 123456
123456
123456
123456 1234567
1234567 123456
123456
50 123456 1234567 123456
123456
123456 1234567
1234567 123456
123456
123456
123456 1234567
1234567 123456
123456
123456
123456 1234567
1234567 123456
123456
123456
123456 123456
123456 1234567
1234567 123456
123456
40 123456 123456 1234567 123456
123456 123456 1234567 123456
123456
123456 123456
123456 1234567
1234567 123456
123456
123456
123456 123456
123456 1234567
1234567 123456
123456
123456 123456 1234567 123456
30 123456
123456 123456
123456 1234567
1234567 123456
123456
123456
123456 123456
123456 1234567
1234567 123456
123456
123456 123456 1234567 123456
123456
123456 123456
123456 1234567
1234567 123456
123456
123456
123456 123456
123456 1234567
1234567 123456
123456 1234567
1234567
20 123456 123456 1234567 123456 1234567
123456
123456 123456
123456 1234567
1234567 123456
123456 123456
123456 1234567
1234567
123456
123456 123456
123456 1234567
1234567 123456
123456 123456
123456 123456
123456 1234567
1234567
123456
123456 123456
123456 123456
123456 1234567
1234567 123456
123456 123456
123456 123456
123456 1234567
1234567
10 123456
123456 123456
123456 123456
123456 1234567
1234567 123456
123456 123456
123456 123456
123456 1234567
1234567
123456 123456 123456 1234567 123456 123456 123456 1234567
123456
123456 123456
123456 123456
123456 1234567
1234567 123456
123456 123456
123456 123456
123456 1234567
1234567
123456
123456 123456
123456 123456
123456 1234567
1234567 123456
123456 123456
123456 123456
123456 1234567
1234567
123456 123456 123456 1234567 123456 123456 123456 1234567
6 7 8 9 10 11 12 13

Date of High Tide MARCH ' 1 9 8 5

Number of adult T.gigas on or near the breeding beach of Chandipur

Fig.9.
Presents the number of spawning horseshoe
crabs T. gigas in two respective high
tides of day and night .
 O
♀ SR
123
123
= 1.05 : 1
123
550 123
123
123
123
123
123
123
123
123
123
123
123 O
123
123
500 123
123
123
123
123
SR = 1.07 : 1 ♀
123
123
350 123
123 123
123
123
123 123
123
O 123 123
♀ SR = 1.04 : 1 123 123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
300 123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123 123 123
123
123 FULL MOON 123
123 123
123
123 123 123
123
123
TIDE 123
123 123
123
250 123
123 123
123 123
123
123 123 FULL MOON 123
FULL MOON 123 123 123
123 123 TIDE 123
TIDE 123
123 123
123 123
123
123
123 123
123 123
123
123
123 123
123 123
123
123 123 123
123 123 123
200 123
123 123
123 123
123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
150 123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
123
123 123
123 123
123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
100 123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123
123
123 123
123 123
123
123
123 123
123 123
123
50 123
123 123
123 123
123
123 123 123
123 123 123
Tachypleus gigas

123
123 123
123 123
123
123
123 123
123 123
123
123
123 123
123 123
123
123 123 123
123
123 123
123 123
123

6 7 8 9 10 11 12 13 Total 1 2 3 4 5 6 7 8 9 10 11 12 Total 31 1 2 3 4 5 6 Total

No of

M A R C H M A Y J U L Y
D a t e in 1 9 8 5
Fig .10
Fig.10
.10. Shows the number of T. gigas counted in successive days of March, May and July,
1985, with respective total of males and females displaying sex-ratio at the tops(this imbalace
idicates increasing death rate of the males in comparism to that of females).
 5820 12345
12345
12345
12345
4000 12345
12345
76
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
3000 12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
12345
Full moon 1234512345
12345
12345
12345
12345
12345
12345 12345
12345
12345 12345
12345 12345
12345
12345 123412345
12345
12345 123412345
12345
12345 1234
123412345
12345
12345
12345 1234
1234 12345
12345 123412345
12345
12345
12345 1234
1234 12345
12345 123412345
12345
12345 123412345 Full moon
12345
12345 1234
123412345
12345
12345 123412345
12345
12345 1234
123412345
12345
12345
12345 1234
1234 12345
2000 12345 123412345
12345
12345 123412345
12345
12345 1234
1234 12345
12345 123412345
12345 12345
12345
12345 1234
123412345
12345 12345
12345
Estimated Population of Tachypleus gigas

12345
1234512345 1234
123451234 12345 12345
1234512345 123412345
12345 12345
12345
12345
1234512345123412345
12345 1234 12345 12345
12345 1234 12345 12345
12345
12345
123451234
123412345
12345 12345
12345
Full moon 12345
12345123412345
1234 12345 12345
12345
12345123412345 12345
12345123412345 12345
12345 12345 1234 12345
12345
12345
12345
12345 1234
1234 12345
12345 12345 123412345
12345
12345
12345 12345
12345 1234
1234 12345
12345 12345 123412345
12345
12345 12345 123412345
12345
12345
12345
12345 1234
1234 12345
12345 12345 123412345
12345
12345
12345 12345
12345 1234
123412345
12345
12345
12345 12345
12345 1234
123412345
12345
12345 12345 123412345
12345
12345
12345
12345 1234
1234 12345
12345
12345
12345 12345
12345 1234
1234 12345
12345 12345 123412345
12345
1000 12345
12345 12345
12345 1234
123412345
12345
12345 12345 123412345
12345
12345
12345
12345 1234
123412345
12345
12345
12345
12345
12345 12345123412345
12345 12345 123451234
123412345
12345
1234
1234 12345
12345 12345
12345 12345
123451234
123412345
12345
1234
1234 12345
12345 12345
12345 12345
123451234
123412345
12345
1234
1234 12345
12345
12345
12345 12345
123451234 12345
1234 12345 12345 123451234
123412345
12345
1234
1234 12345
12345 12345
12345 12345
12345123412345
1234 12345
1234
1234 12345
12345 12345
12345 12345
123451234
1234 12345
12345
1234 12345 12345 12345
12345123412345
1234
1234 12345 12345 12345
1234
1234 12345
12345 12345
12345 12345
123451234
1234 12345
12345
1234 12345 12345 1234 12345
12345123412345
1234
1234 12345 12345 1234 12345
1234 12345 12345 1234 123451234 12345
1234
1234 12345
12345
12345
12345 1234 123451234 12345
1234 12345 123451234
1234
12345
12345123412345
1234 12345
1234
1234 12345
12345
12345
12345 1234
1234 12345 12345
123451234
1234 12345
12345
1234 12345 12345 123412345
12345 12345
12345123412345
1234
1234 12345 12345
123451234 12345
1234 12345 123412345
12345 123451234 12345
1234
1234 12345
12345
12345
12345 1234 123451234 12345
1234 12345 12345 1234 12345 12345
123451234
123412345
1234 12345 12345 1234
1234 12345
12345 123451234
12345
12345
1234
1234 12345
12345
12345
12345 1234 12345 123451234 12345
1234 12345 12345 1234 12345 12345
12345123412345
1234
1234 12345 12345 1234 12345 12345
1234 12345 12345123412345
1234
12345
12345
12345123412345
1234
12345

7 8 9 10 2 3 4 5 6 7 8 9 10 11 30 1 2 3 4 5
MARCH MAY JULY
1 9 8 5

Fig.13.
Showing the number of T. gigas estimated at Chandipur sea shore with
the help of Jackson's "positive" (black bar) and "negative" (dotted bar)
methods.
 77

45 O + ♀

Sex Ratio = 1.41 : 1.00

21P Mated
12345
12345
12345 Solitary
12345
12345 O
12345 160
40 O
12345
Solitary ♀
12345
12345
12345
18P 12345
12345
12345
12345
12345
35 12345
12345
12345
12345
16P 12345
12345
12345
12345
15P 12345
12345
12345
♀
30 12345 120
12345
12345
12345
12345
12345
12345 12345
12345
12345 12345
12345
12345 12345
12345
12345 12345
12345
25 12345
12345 12345
12345
12345 12345
12345
12345 12345
12345 12345
12345
12345
12345 12345
12345
12345
12345 12345
12345
12345 12345
12345
12345 12345
12345 80
20 12345
12345 12345
12345

No of Solitary Animals
12345 12345
12345
12345 12345
12345
12345 12345
8P 12345 12345
No of Mating Pairs

12345
12345 12345
12345 12345
12345
12345
12345 12345
12345
15 7P 12345
12345 12345
12345
12345 12345
12345 12345
1234
1234
12345
12345 12345
12345
12345 12345
1234
1234
1234
1234
12345
12345
12345
12345
1234 1234 5P 12345 12345
12345 12345
1234
1234 1234 12345 12345 40
10 1234 12345
12345
1234
1234
12345
12345
12345
12345
12345
12345
12345
12345 1234
1234 12345
12345 1234
1234 1234
1234
12345
12345
12345
12345
12345 12345
1234 12345
12345 12345 1234
1234 12345
12345 1234
1234 1234
1234
12345
12345 12345
12345
1234 12345 12345 1234 12345 1234 1234 12345
12345 12345
12345
1234
1234 12345
12345 12345
12345 1234
1234 12345
12345 1234
1234 1234
1234
12345
12345
12345
12345
5 1234 12345 12345 1234 12345 1234 1234 12345
12345
12345
12345
1234 12345 12345 1234 12345 1234 1234 12345
12345
12345
12345
1234
1234 12345
12345 12345
12345 1234
1234 12345
12345 1234
1234 1234
1234 12345
12345
12345
12345
1234
1234 12345
12345 12345
12345 1234
1234 12345
12345 1234
1234 1234
1234
12345
12345
12345
12345
12345 12345
1234 12345 12345 1234 12345 1234 1234 12345 12345
1234 12345 12345 1234 12345 1234 1234 12345 12345

March April May June July Aug. Sept. TOTAL

1 9 8 7

FIG.11.

Represents the occurrence of mating pairs (P) , solitary males and

females of Carcinoscorpius rotundicauda , and showing the sex- ratio
(SR) at the extreme right hand column .
Mated males and females are very hard to separate
from the amplexus and when the two are separated
forcibly and placed on sand or hard mud-bed the
larger ( female ) one moves down toward the lower
littoral zone which the smaller (male) appears to be
helpless and does not usually follow its mate . But
placed close enough , however , the male follows the
female's track (Fig. 25) to get its normal position .
This field observation was also endorsed by Drs. B.N.
Chopra and H.S. Rao (c c.f. Roonwall , 1944).

From the capture-recapture data and from the collection

of dry carapaces male to female sex-ratio was found to be
6.89:1 at Digha and 1.04:1 at Chandipur . This disbalance
in normal sex-ratio (1:1) .................................................
at Digha fishermen enhanced the predation by throwing
them to crows . In Thailand disbalance in sex-ratio of T.
gigas and C. rotundicauda is also noticed which is due
to the fact that the people collect the eggs to serve on the
table as food (Sekiguchi et. al. , 1977 : Sekiguchi and
Nakamura , 1979 and Dr. A . Choudhury's personal
observation on 1986) . Another possible factor
...............................may be the "unauthorised sacrifice"
of the female crabs ( because of their large size) as very
important laboratory material for bio-medical research by
draining its ' blue blood' to get the precious lectin
"carcinoscorpin " (Dorai et. al. , 1981 ; Srimal et. al. ,
1985 ; Vijaya Rao and Bhagirathi ,1989; Rudloe , 1983)

ø W hy the wor
wor ds lik
ords e "unauthorised sacrifice" ?
like
..............asked the foreign expert.............. for explanation please see p.72
 79

DISCUSSION :

Population of the American horseshoe crab Limulus polyphemus was estimated following the capture
- recapture methods of Baptist et. al. (1957) , Rudloe (1980) , Shuster (1950) and Sokoloff (1978) . For
the first time in the Indian sub-continent Debnath and Choudhury (1988a) estimated the population of one
of the two Indian horseshoe crabs, Tachypleus gigas by using the above principle at Chandipur sea
shore .
The breeding horseshoe crabs ( T. gigas ) at Chandipur showed strong lunar and tidal rhythms , with
animals appearing at and within few days of fullmoon and within 2 hrs. of the time of flood tide(Fig.
9).Among several variables only lunar cycle , day of the year , and wave- height correlated significantly
with number of carbs on the beach (Rudloe , 1980) . Table 20 shows the significant correlation between
total marked (after initial capture) animals and recaptured animals in March , May and July . Total recov-
ery rates of recaptures in respect to the total captures of each month are 11.7% , 27.41% and 15.44%
respectively . These results indicate that the maxima of population occurred in May i.e. , the mid-breeding
season and which started in February - March and declined in July-August (Fig. 10) . Considering the
recovery rates in term of sex this table also showed that the males always dominated over the females .
This kind of male sex dominance over the females was common in L. polyphemus , but expected to be a
normal sex-balance in Indian species . From the capture-recapture data and from the collection of dry
carapace , male to female sex-ratio was found to be 6.89 : 1 at Digha and 1.04 : 1 at Chandipur
(Debnath , 1985) . This abnormality in normal sex-ratio (1:1) seemed to be a result of severe predatory
pressure at Digha than at Chandipur which was caused by C. splendens ( Debnath and Choudhury ,
1988a) .
Furthermore , the capture-recapture data indicates a clear variation of the total number of arriv-
ing animals on or near the breeding habitat . The maximum numbers occurred on 9th March (5,820 ani-
mals) . The author , however , failed to capture horseshoe crabs on the following new moon tides (15th to
25th March) . This was also evident in the new moon tides of May and July . Population peak was also
recorded between the dates of 4th and 9th May , associated with the full moon tides of 6th , while in
July , maximum number of crabs were estimated on or before 5th , the full moon night (Fig. 13) .
For animals , dispersion is a common phenomenon in case of mobile species and it has also
been proved from the result of the capture-recapture experiment performed at Chandipur sea shore within
an area of 2 X 3 sq. Km. But an interesting observation in recovering two segregated opposite sexes
further amplexed (coupled) 3 days after release has rendered the author to assume that specific sexual
selection might had occurred during the breeding activities. But how they choose themselves and found
together and returned to the breeding ground remained a great mystery .
Direct application of Lincoln - Peterson Index on 16th -17th March 1986 at Chandipur re-
vealed 4,260 animals that came to breed which in turn has been corrected as 4,071 with variance and
standard error SE ( + 835.439) following Bailey (1951 , 1952) . This observation corrobarates with the
findings of the same month of the previous year (1985) following Jackson's principle .
While estimating the population size at Digha sea shore , the errors that evolved in Jolly-Seber's
stochastic method may be the result of two factors ; a) "small sample biases" inherent in the method and
b) biases due to the violation of the assumptions. The former will be minimal when the number of recap-
ture is large . Manly (1971) and Roff (1973) have shown that these errors often express themselves as
estimates and their variances . Some improvements could be obtained if N and O are transformed to
logarithms before making the calculations ( Manly , 1971 ) . If one looks at Table -17 on the row of 18th
May , it becomes clearly evident that error due to "small sample biases" crept
 80

^N (457.142) ^
i) 18 < VN18 (524.197)

^N ^ / ^
ii) 18
(457.142) < VN 18
N18 (523.760)

on that very day (18th May) a total 56 animals were marked and released in which the number of
recapture ( marked on 17th) was only 2(two) and the survival rate ,^
θ18 is so low (0.0914) that all results
gave considerable errors .
However , Jolly-Seber's method (1965) applied for the estimation of T. gigas population at
Digha in the year of 1986 showed variation in the estimates (120.5 to 1703.6 ). Maximum number of
animals occurred at the full moon tides . Numbers of emigrated or immigrated animals were also deter-
mined with their variances (Table - 17) .

Discredit should be given to the

discrete local population of Indian
horseshoe crab, T. gigas at the sea
shore of Digha as because the
animals were being repeatedly
threatened by human interference
, continued fishing with long tow
nets and over-predation by Corvus
splendens (Debnath , 1985 ;
Debnath and Choudhury , 1988a) .
 81

D. Absolute population estimates of Tachypleus gigas with regression lines:

In the foregoing chapters the author has discussed various marking techniques to estimate the popu-
lation size of T. gigas in which there were distinctively three sets of data i.e. i) estimate popula-
tion (N) , ii) capture , marked and released (C) and iii) recaptures (R) . Here the author has deployed
both correlation and regression between N vs. C and N vs R . Now , theoretically , if in an imaginary area
of estimate , the number of total captured , marked and released animals become equal to recaptured
animals the area , obviously , will show the absolute estimate (N) . This hypothesis can be obtained from
the " Lincoln Peterson Index " as below :
nxa
N = , i.e. N = n , when a = r
r
Keeping this in the mind the author has constructed two regression lines and placed them on graph
paper . The co-ordinate on which two regression lines intercept each other will indicate the X-value for
C = R and thus the Y value for absolute estimate (N) of the population .

RESULTS
Correlation coefficients were determined between N & C and N & R for both Jackson's
and Jolly-Seber's methods , with its standard errors (se) and t-values (Table 19) . Then two regression
lines were constructed for both the methods and fitted them on graph papers (Fig. 14) with intercepting
co-ordinates and tangants to X-axis . The intercepting co-ordinates ( X and Y-values) were then antilogged
to find out the theoretical estimates .

E. Population estimation of standared T. gigas at Chandipur beach by Line Transect method

In the soft sandy shores of Chandipur , T. gigas , mostly in pairs , remain scattered partly or
fully embedded under silty-sand to avoid desiccation and rising temperature , and its anticipated predators
Corvus splendens . The alternative explanation for these stranded "crabs" may be placed like this that to
avoid to and fro repeated journey between the tide marks during the ebb , they prefer to stay at the upper
shore level embedded under the substratum only to wait for the arival of the next tide . An attempt was
taken by the author to estimate the population of these standard and sparsely distributed carbs in an area
of 1 Km2 of Chandipur from 29th March to 31st March , 1986 following Line Transect method (Vide
Chapter : Materials and Methods) . Every day eight transects of 10 m2 each were randomly selected on the
stretch of 1 Km to get the count of the crabs .

CALCULATION AND RESULTS

Horseshoe crabs counted in each transsect (Table 18) are calculated in two ways :-
1. Population of T. gigas in 1 sq. Km. in terms of area (ITOA) .
Mean value of N = 3437.5 + 1541.52
2. Population of T. gigas in 1 sq. Km. in terms of days (ITOD) .
Mean value of N = 3437.5 + 499.56
Now if this mean value of N is campared with the estimate (followed by Lincoln's method) of
16th - 17th March the t-test did not show a significant deviation as found the comparison of two means
described below :
 82

Comparison between two means :

Capture-recapture method Line Transect method
( 26th - 27th March , 1985) ( 2oth -31st March , 1986)

n = 4 12
X = 2142.25 3437.5
SD = 2127.958 1541.52

X 1 - X2
t = = 1.123, df. = (12+4) - 2 = 14 , p 0.05
S2 1 S2 2
+
n1 n2

Total 5000 and 7500 horseshoe crabs were estimated for 26th and 27th March , respectively
by using the line transect method (Table 18) and the average value thus obtained is 6250 . Then following
Gaskell and George's (1972) formula
an + 2N , where N is the estimate followed after the line transect method .
n = r+2
300 X 284 + 2 x 6250
=
20 + 2

= 4,441 .
Therefore , the population estimate of T. gigas following the three different principles comes to :

1)Lincoln - Peterson Index (1930) : N = 4,260 + 918.41

2)Bailey (1951 , 52) : N = 4,071 + 835.43
3)Gaskiell and George (1972) : N= 4,441

The above figures of N indicate that the population of T. gigas at Chandipur sea shore was
nearly 5,000 that came to breed in the month of March , 1986 and this figure corroborated with the
maximum estimate (N= 5,820) of the same month i.e. , March of previous year (determined after
Jackson's formula) .
 83
5.0
4.85 ( N = 70,794 )
Field Station :
CHANDIPUR ( ORISSA )
6
.655
=0
4.0
5
9,
r
647
25 0.
2.2
Fig . 14
r =
X
+
54,

0.6
608
1.
28 Absolute
+
3.0 Y= 2
X population
882
(Ñ)

0. of T. gigas
Y=
ESTIMATED POPULATION

∆ = 33.45
o estimasted with
regression method
2.0
at two different
field stations
∆ = 41.42
o
Chandipur
and
1.0
Digha

0
1.0 2.0 3.0 4.0
CAPTURE / RECAPTURE
3.5
3.26 ( N = 1820 ) 1
Field Station : 0.481
=
2, r
2.287
DIGHA 21 X
+
(West Bengal) 3.0
=0.36
872
Y 6
0.
r =
09,
07
∆ = 19.90
o 1.
(Ñ)

+
X
78
.85
ESTIMATED POPULATION

0
=
2.5
Y

o
∆ = 40.62

O 1.5 2.0 2.5

CAPTURE / RECAPTURE
 84

DISCUSSION

At Chandipur for one breeding cycle during the period of March to July , 1985 , both correla-
tion and regression showed significant results (Table 19) where the maximum likelyhood or theoretical
number of capture , marked and released animals (C) became equal to recaptures (R) , i.e. , 8,912 . In that
case the author can predict that maximum 70,795 animals (Fg. 14) came to breed at Chandipur in 1985
during the spawning season (spring and summer months) .
While at Digha , the correlation between N and R showed non-significant relationship ( r =
0.481 + 0.213 , P> 0.05) , but significant correlation between N and C (r = 0.687 + 0.146 , P< 0.01) .
Thus if the non-significant former regression line intercepts with the significant latter line the X value
showed 283.79 and Y = 1820.00 . That means almost 1500 T. gigas came to breed at Digha in May
1986 . But this may be an underestimate because of insufficient data regarding recaptures for which" r"
showed non-significance in the relation between N and R .
The author , in this context , surmises that a complete set of date of one fullmoon breeding
season could have yield better result as in the case of Jackson's method applied for Chandipur popula-
tion . It has already been established in the field of population estimation that Jolly-Seber's stochastic
model is the only deterministic approach if it becomes fitted with a complete and unbiased data
(Southwood , 1978) . Discredit should be given to the discrete local population of T. gigas at the sea
shore of Digha as because the animals were being repeatedly threatened by human interference , contin-
ued fishing with long tow nets and over-predation by Corvus splendens (Debnath , 1985 ; Debnath and
Choudhury , 1988a) .
During the feild investigation of the project programme there were severe constraints
regarding field infrastructure facilities in the remote coast line areas of the two states (Orissa and West
Bangal) . The author was quite aware of these short coming and tried his level best to bring down the
apprehended statistical errors during the generation of data related 'crab' population dynamics . What the
author contends is that an organised team with working facility in the feild and assistance for a
longer duration taking many more stations amost the coast may help to bring out a more comprehen-
sive population picture of the Indian horseshoe crabs .
 85

D . Breeding activity ; Sex-ratio and predation

 86

INTRODUCTION

The mating or spawning behaviour of all extant horseshoe crabs , comprising three genera and four
species , is one of the easiest facets of the Limulidae life cycle to study because the animals come ashore
to spawn . Although their breeding behaviour is remarkably similar (Shuster , 1982 ; Sekiguchi , 1988) .
Detailed study on the breeding behaviour and spawning activity of American horseshoe crab L .
polyphemus have been conducted by various workers as because of its greater abundance and wide range
of spawning areas (Barlow et. al . , 1982 , 1986 ; Botton et. al. 1988 ; Brockmann , 1990 ; Cavanough ,
1975 ; Cohen and Brockman , 1983 ; Dunton , 1953 ; Finn et. al. 1990 ; Rudloe , 1980 ; Rudloe and
Herrnkind , 1976 ; Shuster , 1953 , 1958 ; Shuster , 1979 ; Shuster and Botton , 1985) .
Despite the occurrence of three species of horseshoe crabs , widely distributed in the coastal waters
of south-east Asia , very little information is available on their breeding behaviour , spawning migrations
and other activities in comparison with that of L. polyphemus .While, Sekiguchi and Nakamura(1979),
for the first time, provided a comprehensive and comparative account of Asian horseshoe crabs with
reference to their breeding biology . In case of the Indo-pacific horseshoe crabs , the breeding season
also varies geographically , from species to species (Debnath , 1985 ; Debnath and Choudhury , 1985 ;
Sekiguchi , 1988 ; Sekiguchi and Nakamura , 1979 , Waterman , 1953) .
 87

OBSEVATIONS AND RESULTS

A. Breeding season :

In the present study the breeding season of Tachypleus gigas has been found to vary from late
February or March to July . On the other hand , Carconoscorpius rotundicauda arrives in the estuaies ,
especially in Sundarbans (Prentice island and Sagar Island) from March to September . While , in the Gulf
of Siam (Thailand) this species lays eggs all round the year (Sekiguchi & Nakamura , 1979) . It is also
generally recorded that the egg laying season begins from April both in T. gigas and C. rotundicauda .

B. Spawning activity :

T. gigas and C. rotundicauda are found in the coastal waters of India , inhabiting the Bay of
Bangal , from the sandy shores of Orissa to the muddy-sands of estuarine Sundarbans , West Bengal . Both
T. gigas and C. rotundicauda usually make pair in the open water in the infra- littoral zone prior to
reaching the spawning area (Debnath , 1985 ; Debnath and Choudhury , 1988a) .

During the breeding season horseshoe crabs emerge from the sea and come to the shore line upto the
highest mark of high tide associated with fullmoon tides in mating amplexus (Fig. 22 A,B) . The females
carries the male on her back , the latter is being attached to the female's opisthosoma by holding her
marginal spines with his modified clasper legs (Fig. 22 B , 16A) . In sexually mature males , the 2nd and
3rd prosomatic appendages (1st and 2nd legs) are modified into clasper organs . The male clasper organ is
chelate hook-like in C. rotundicauda , while it is non-chelate , bent and stout hook in T. gigas(see
Fig.18 C) . The holding mechanism is so strong enough that to separate the male from the female's back
is very difficult .

At Chandipur (Burabalanga river mouth) the breeding activity of T. gigas has been studied in great
detail . It has been observed that the animals come near the sea shore in pairs (amplexed) when the tide
level reaches at the upper littoral zone . The pairs are characterized by their larger female partner carrying
a male of small size on her back . Mated pairs are usually found to prefer the shore where wave action is
minimum . The high tide peak stayed for 20-30 mins . during which time , the amplexed crabs showed a
kind of sexual courtship just along the margin of the tide water . All of sudden one pair may emerge from
the water's edge , move freely here and there , then disappear in the water and reappear nearby or at a
distant place , possibly in search or selection of a suitable site for spawning .

On the exposed intertidal beach , when the water receeds back , some characteristic horseshoe shaped
swollen impressions on the sand helped the investigator to detect the mating couples . Such markings
were indistinct on the mud flats of Sundarbans (West Bengal) and Dhamra (Orissa) due to the presence of
heavy mud enriched with silty clays . In that case somewhat depressed area with broken mud cake re-
vealed the existence of C. rotundicauda ( Fig. 21B , 22c) .

Studies with mechanical marking as well as tagging with rubber bands on the seggregated males and
femals of both species revealed no further formation of amplexus or coupling between opposite sex on
or near the beaches of sand- or mud-shores except an exceptional occurrence at Chandipur (Table -20) .
This is an indication that the crabs , having spawned on one beach , did not spawn on another.
 88

C. Sex -ratio :

During high tide pairs of T. gigas were captured from the Burabalanga river mouth . Animals of
both species of Indian horseshoe crabs showed 1 : 1 sex- ratio during pairing . But at the low tide when
the intertidal sand or mud-flats get exposed showed some sort of disbalance in the above sex-ratio.
Because it was observed that in comparison to that of "solitary females" the solitary males are "very
common". Since the total capture-recapture data included both the samples of high tide and low tide ,
the observed results obviously revealed the presence of higher number of males over the females in the
spawning area ( Fig. 10) . Similarly, the total net collection of T. gigas at Digha sea shore usually showed
more males than females . Solitary males were very common at the mud-flats of Prentice island ,
Sagar Island (Chemaguri ) and Dhamra (Koithkola, Orissa) . In all the above study sites C. rotundicauda
occurred except at Dhamra where both T. gigas and C. rotundicauda orrurred in the same creek (Fig.
12).

D. Predation by crows , Corvus splendens L. : (Debnath & Choudhury , 1988a)

At Chandipur and Digha several mating pairs of T. gigas were found on exposed beach (fig. 27 A)
during low tide when they were unable to make burrow quickly and hide themselves in the sand( Fig. 27
B) . It was the appropriate time for the predator birds to locate their prey . The crow sat beside the pair and
upturned them with their beaks and claws . The crab always tried to protect itself by moving the sword-
like telson around . But once they were uoverturned , they were susceptible to predation easily , espe-
cially the female crabs . The predator pierced the ventro-lateral soft chitinous layer of females prosoma
and took eggs one by one , while the male still remained clinged to the marginal (opisthosomatic)
spines of the female with his clasper legs . In most cases it was observed that the male remained unhurt by
the crow and the predator hardly took the flesh of the crab , although injured by mistake ( Fig. 27 C) . It
was also observed that more than one crow shared the paired crabs , but normally one crow was found to
be engaged in this operation ( Fig. 27 D) . A gravid female usually carried 100 to 180 gm of eggs in an
average of 600 gm of total body weight . The whole prosoma was being filled with mature eggs , (in an
average 6,000 in number ) . Approximately , each egg measuring about 3.7 - 4.0 mm. in diameter and
looked like the seeds of the ladies finger (Fig. 28 A) .

E. Tandem amplexus in C. rotundicauda :

During June 1988 , the investigator found 5 to 25 mating pairs of C. rotundicauda in each of
the seven narrow channels (sub-creeks) of Chemaguri Creek situated at the south-eastern part of Sagar
Island , West Bengal . While stdying the breeding behaviour of these pairs ( Fig. 23 A) it has also been
observed that an unusual coupling of two males , one behind the other (in tandem) . When the mud was
cleared away , in front of the forward male , a female was exposed (Fig. 23 C) . All these three animals
were in a linear row (Fig. 23 B) . When pulled out of the mud and the manner of attachment examined , the
first male was found to be attached to the female , hence a "clasping" male , with the second male attached
to the first male , forming a "satellite" * . These two males were about equal in size , shape and weight .

Table 21 presents the total counts of male and female horseshoe crabs of both species ( T. gigas
and C. rotundicauda) and their sex-ratio (SR) obtained from five stations , ranging from estuarine
Sundarbans of West Bengal to Dhamra of coastal Orissa 6.89 :1 was the highest SR determined at
 89

Digha , while the lowest one was 1.04 : 1 obtained from Chandipur ; in both the cases the studied
specimen was only T. gigas . Both T. gigas and C. rotundicauda have been found to occue in creeks
and mud-bottoms of estuarine Dhamra as sympatric population , where they showed almost similar
sex- ratio. The X2 - test of male versus female horseshoe crabs reveals that it is being significantly
disalanced in the three coastal stations of West Bengal , amongst which Digha sea shore showed the
most vulnerable area for severely damaged sex-ratio (X2 = 83.62 , P< 0.001) . On the other hand , the
coastal range of Orissa was not as that much under predator pressure to reverse the desired sex-ratio .

A combined histogram (Fig. 12) has been presented for showing the occurrence of horseshoe
crabs in two mud - flat mangrove ecosystems of Chemagari and Dhamra , respectively in W.B. and
Orissa . Total male and female counts revealed a high deviation in sex balance of C. rotundicauda in
the former station (SR = 1.44 : 1 , P= 0.01) , while the SR (1.09:1) for the same species in Dhamra
showing no significant deviation i.e. P> 0.05 . such is the case for T. gigas (SR = 1.21:1, P> 0.05) at
Dhamra .

SOLITARY
CRAB

PAIRED / MPLEXED
CRABS
 90

Table 21 .

Study area , period , total males and females of C. rotundicauda are T. gigas are
shown to determine the male to female sex-ratio and Chi-square (X) tests* with
significant (P) levels

2
Study Area Period Species type Total Total Sex Ratio X - test P
Male Female (Male/Female)

Prentice Island Feb'86 to C. rotundicauda 218 177 1.23 :1 4.255 0.05

West Bengal Nov. '87 (197.5) (197.5)

Chemaguri Sagar March -June C. rotundicauda 296 205 1.44:1 16.528 0.001
Island , W.B. 1988 (250.5) (250.5)

Digha Sea Shore May-June T. gigas 131 19 6.89:1 83.626 0.001

West Bengal 1986 (75) (75)

Chandipur Feb - March T. gigas 333 320 1.04:1 0.258 ------

Orissa 1985 (326.5) (326.5)

Dhamra April-May T. gigas 75 62 1.20:1 1.233 ------

Orissa 1987 (68.5) (68.5)
C. rotundicauda 69 63 1.09:1 0.272 ------
(66) (66)

*Theoretical values in parenthes is below the observed values

A bunch of Eggs seems to be the bunch of Pearls

Table 22 . Movement , dispersion and recovery rate (R) or the breeding populations of two Indian horseshoe crabs at four coastal stations of Bay of
Bengal , India .

Study Area Species type and Number of Number of Marking device Release of Release of Recovery of Recovery of Remarks
period intact pairs seggregated intact pairs seggregated pairs : number seggregated
pairs at distance animals at and distance animals ; if
from the site distance from any
of capture one another

Chandipur sea shore T. gigas 50 15 Mechanical markings 50-200 mts 5-20 mts 2 at 5 mts 1 pair , both Active
Orissa (High tide) (March-July) (with steel needle) on apart from 2 at 15 mts sexes with similar
1985 carapaces one another 2 at 50 mts marks recovered
1 at 100 mts at 200 mts away
1 at 200 mts from the orginal
1 at 1000 mts site of capture
Total = 9,R = 18% R = 6.66 %

Digha sea shore T. gigas - 10 Do Do Do Nil Nil Active

West Bengal (May-June)
(High tide) 1986

Koith-Kola , Dharma T. gigas 25 10 Colored rubber bands Manipulated Random release Total =8( R=32% ) Nil Sluggish
Orissa (Low tide) tied on telson trunk released in I-II in I & IIsub =10 (R=40%)
C. rotundicauda 25 10 sub creeks creeks Recovered from sub-
(April-May) (ave. distance creeks and mud flats
1987 200 mts.) with random distribution.

Chemaguri , Sagar C. rotundicauda 50 15 Do Manipulated Random release Total 17 pairs Nil , but with 6 Sluggish
Island , W.B. (May -June) released in I in III - IV sub- recovered from similar marked
(Low tide) 1988 to V sub creeks creeks sub-creeks & males & females
(ave. distance mud flats with in same creek (IV)
100 mts.) random distri distributed with in
bution R=34% 50 mts .
91
 92

A quack
practitioner
sacrificed
the crabs,
cut off the
limbs, boiled
in mustard
oil and sold
it to the
people !

possible factor for the

depletion of the female
horseshoe crabs in coastal
West Bengal of Orissa
may be the unauthorised
sacrifice of the female
crabs ( because of their
large size)

12345
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SR= 1.09 : 1
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SR= 1.21 :1
Counting of horseshoe crab population at field station,DHAMRA,Orissa 12345
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1 2 3 4 17 18 19 20 21
APR I L' 1 9 8 7 MAY ' 1 9 8 7
Fig.12 A. Histogram showing the total number of C. rotundicauda at DHAMRA, Orissa
with respective sex-ratio (SR)
 100 200 300
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2 3 4 18 19 1 2 3 16 17 18 31 1 2 3 16 17 31 1 2 3 15 16
MARCH APRIL MAY JUNE
1 9 8 8
Fig 12 B. Histogram showing the total number of C. rotundicauda at Chemaguri ,
West Bengal with respective sex-ratio (SR)
 94
DISCUSSION

The differential sex-ratios in the population of the Indian horseshoe crabs in the coastal
West Bengal and in coastal Orissa (Table 20) may underline the following ecological consequences :

i) In comparison to the coastal West Bengal , people of Orissa were found to be more conscious
about these marine creatures and attaching some religious superstition on the pairing of the 'crabs' as
"Rama-Laxman" the mythological two brothers of the holy Epic the Ramayana and hence unknowingly
helped in their partial conservation ,
ii) More and more Anthropogenic interference at the sea resorts of Digha , Junput , Bakkhali ,
Frazerganj , Sagar Island and other places of coastal West Bengal inflicting random habitat destruction ,
alarming sea resource exploitation , and others may be the major causative factors for disbalancing the
sex-ratio,
iii) Natural predation by Corvus splendens is common everywhere , but this predation pressure is
enhanced manifold by the fishermen at Digha as they throw the 'crabs' to the exposed sandy beach believ-
ing them as enemies to their fishing nets ,
iv) Furthermore , the largest freshwater draining rivers like Ganga , Matla and several others carry
a vast amount of chemical pollutants , industrial effluents , oil spills and a variety of toxic substances
( Botton & Loveland , 1989) into the estuarine Sundarbans which may severely affect the normal sex
balance and fecundity of Indian horseshoe crabs in coastal Bengal than in coastal Orissa , and finally,
v) Although the local people do not eat the eggs of horseshoe crabs like the people of Thailand
(Sekiguchi and Nakamura , 1979) , the female specimens are much preferred by the animal traders
(quacks , Debnath, 1987) due to its large size .
The present chapter communicates the first report on the predation of horseshoe crabs ( T. gigas) by
the shore birds ( Corvus splendens) in Indian sub-continent (Debnath and Choudhury , 1988a) . A unique
prey-predator relationship was observed in coastal Orissa (Chandipur) and West Bengal (Digha) which
revealed that population of crows increased at the time of breeding time of the 'crabs' . Predator birds have
reported to arrive at the Cape Shore within a week or two with the onset of Limulus polyphemus
spawning in May and generally depart by July ; by this time Limulus abundance has been found to decline
and most eggs of the horseshoe crabs were hatched into trilobite larvae (Botton , 1984c) .
From the capture-recapture data and from the collection of dry carapaces (Fig. 28 B) male to female
sex-ratio was found to be 6.89:1 at Digha and 1.04:1 at Chandipur (Table-21) . This disbalance in normal
sex-ratio (1:1) seemed to be a result of severe predatory action at Digha than at Chandipur as in the later
case crows get access to the 'carbs' in natural way , but at Digha fishermen enhanced the predation by
throwing them to crows . In Thailand disbalance in sex-ratio of T. gigas and C. rotundicauda is also
noticed which is due to the fact that the people collect the eggs to serve on the table as food (Sekiguchi et.
al. , 1977 : Sekiguchi and Nakamura , 1979 and Dr. A . Choudhury's personal observation on 1986) .
Another possible factor for the depletion of the female horseshoe crabs in coastal West Bengal of Orissa
may be the "unauthorised sacrifice" of the female crabs ( because of their large size) as very important
laboratory material for bio-medical research by draining its ' blue blood' to get the precious lectin -
"carcinoscorpin " (Dorai et. al. , 1981 ; Srimal et. al. , 1985 ; Vijaya Roa and Bhagirathi ,1989;
Rudloe , 1983) .
The breeding activity of Indian horseshoe crabs , particularly T. gigas has been studied readily at
high tide time at Chandipur . In and around full-moon day-night observations (March , 1985) it was
revealed that the number of mating animals reached its peak during the day tide than night tide (Fig. 9) .
No solitary animal for mating pairs could be recovered during the approaching new moon tides .
 95

Similar observations were made in May and July in the same year . From the whole result (Fig. 10 ) it
becomes apparent that the mid-breeding time approximated at May when maximum population came to
brred at Chandipur . The histogram results (Fig. 10) also indicate that the sex-ratio (SR) increases very
slowly from the start to the end of spawning like 1.04:1 in March to 1.05:1 in May and finally 1.07 :1 in
July .
Recovery of mating pairs as well as solitary individuals of either sex from the exposed sandy
beach or muddy substratum have ecological bearings . like i) the animals have tried to shorten their to
and fro-journeys at the coast of each and every high tide and ebbe tide , ii) hide themselves from the
predators partly or fully submerging (Fig. 22 C) in the sand or mud , and iii) protect themselves from
desiccation , rising temperature , sun-burn and oxygen defieiency in the gill-flaps due to loss of mois-
ture . Onece these temporarily buried animals were thought to be as egg-laying " crabs " , but no eggs were
obtained from such burrows in the intertidal exposed beach (Fig. 20 A , 21 A) . However , a soft pressure
at the base of the genital apertures situated undersurface to the genital operculum ejaculated eggs one by
one (Fig. 21 C) and milting sperms . Similarly , a sizeable number of L. polyphemus buried in the
intertidal beach throught the subsequent low tide which return to the beach to breed again on the following
high tide (Rudloe , 1980) .
The mating activity of horseshoe crabs is related to the environmental changes produced by
periodic motions of the earth and moon (Barlow et. al. 1986) . Annual orbital movement of the earth
around the sun produces seasonal changes in day lengh and in ocean temperature . While the daily
rotation of the earth about its own axis also creates diurnal changes . On the other hand the monthly
rotation of the moon around the earth modulates the tidal amplitudes of the ocean and produces periodic
inequalities in the daily tides in the northern and southern hemispheres . All these astronomical events
affect the physical environment of intertidal zones , and for the above reasons horseshoe crabs , T.
gigas and C. rotundicauda come to shore during spring and summer months associated with full -
moon tides only . Though a sizable number of solitary males and few pairs of C. rotundicauda have
been obtained from the exposed mud-flats of Prentice island , Chemaguri and Dhamra . Such shoreward
migration was also reported for L. polyphemus during full-moon tides (Teale , 1957 ; Rudloe , 1980).
She (Rudloe , 1980 ) also reported maximal mating activities during full-moon high tides , but without
any breeding activity during neap tides . However substantial mating also occurred
µ during new moon ,
specially at the begining of the season when overall numbers were large . Cohen and Brockmann
(1983) instead , reported the arrival of more animals at the seashore Bay (Florida) in the same Atlantic
coast during the full-moon only for breeding purposes . Again , Cavanaugh (1975) , and Barlow et. al.
, (1986) observed the arrival of horseshoe crabs during both the full and new moon tides . On the
contrary , no mating activity has been reported from the Indian coast during new moon (Fig. 10 & 13).
A study of neuroplysiological light intensity test on Limulus median ocellus (Lall and
Chapman , 1973) revealed the sensitivity to illumination as low as 1.4 X 10-10 w/(Cm2. nm) at 360
NM , whereas the moon-light intensity measured by Kampa (1970) is well within the range of adult
horseshoe crab's vision . Beside those factors , tide height and sunlight ( as it varies according to the
days of year) were also correlated significantly with the migration of Limulus towards the shore
(Howard et. al. , 1984 ; Barlow et. al. , 1986) . Seasonal variation in Limulus visual sensitivity was
determined with both physiological as well as behavioral studies which indicated the sensitivity was
highest in the summer new moon , while lowest in the winter full-moon (Ross Farhang et. al. , 1990).
 96

A comparable lunar breeding patterns for the two Indo-pacific horseshoe crabs ( T. gigas
and C. rotundicauda ) have been reported ( Sekiguchi et. al. , 1977 ; Debnath , 1985 , Debnath and
Choudhury , 1985 ; Choudhury and Hafizuddin , 1980) . Tagging study with rubber bands as well as
mechanical marking (Table -22) has revealed the following facts :-

a) T. gigas prefers the sand shore having potentially low wave action and comes in the same
spawning ground day after day during full-moon only . But C. rotundicauda prefers the muddy shore
associated with mangrove vegetations (Fig. 20 A-C , 26 A-C) .

b) Dispersion is a common rule in T. gigas , while C. rotundicauda tends to remain in the same
breeding ground .

c) Amplexus occurs in the off-shore shallow water but not in the mud or sand-beach . This inference can
be made from the recovery of males and females in isolation bearing similar marks/rubber bands days
after capture , tagging and segregation .

d) Mated males and females are very hard to separate from the amplexus and when the two are separated
forcibly and placed on sand or hard mud-bed the larger (female) one moves down toward the lower littoral
zone which the smaller (male) appears to be helpless and does not usually follow its mate . But placed
close enough , however , the male follows the female's track (Fig. 25) to get its normal position . This
field observation was also endorsed by Drs. B.N. Chopra and H.S. Rao (c.f. Roonwall , 1944) .

Cohen and Brockmann (1983) have classified the breeding activity of the males and three
catergories : (1) Clasping - the male grasping the posterior opisthosomal angles of a female ; (2)
Satellite - a male not clasping a female but within 10 cm of her or of the males eigher clasping her or
also satellite to her , and (3) Solitary - a male neither clasping nor satellite . While this and other
studies (eg. Rudloe , 1980 ; Brockmann , 1990) have been conducted along the shores of the Gulf of
Mexico , the behaviour is essentially the same in other populations. Thus , the main focal point of this
chapter , tandem amplexus was reported as early as 1870 by the Rev. S. Lockwood .

The first record of tandem amplexus in C. rotundicauda (Debnath and Choudhury , 1992b
appears to be the equivalent of the behaviour reported by Lockwood (1870) and others like Cavanaugh
(1975) for L. polyphemus . The difference , if any , would be when and where amplexus took place ,
relative to the breeding area . In the American species there are generally more males than females on the
breeding beaches . Also , those males that have not found a mate tend to proceed to the foot of a beach and
await the arrival of an unattended female . Attachment occurs when a female moves up the beach slope to
the water's edge (Shuster , 1953) . Extra males may accompany the pair onto the beach and participate in
the spawning process . Usually most of the males , attached or satellite , are milting profusely . The
relative contributions (of sperms ) of all of the males in a spawning group have been analysed by
Brockmann (1990) .

In the case of C. rotundicauda , where extra males are not common and where amplexus
occurs prior to the adults reaching the breeding area , it is surmised that the tandem attachment reported
here probably occurred off-shore (Debnath and Choudhury , 1992b) .
 97

Fig . 20 ( A-C)

A , Mating pair of C. rotundicauda embededv party on the intertidal soft mud

bottom of Prentice island ; B , Upturn posture of the same showing typical
amplexus , C, Shows a morphologically injured female of C. rotundicauda being
clasped with the normal male .

C
 98

Emergence of Eggs of
eggs through C.rotundicauda
genital pores (enlarged)

C Eggs of T.gigas (enlarged)

Fig . 21 ( A-C)
A , The female "crab" carrying a male on her back fails to makes burrow
in the hard mud-bottom ; B , Shows a clear depression in the mud with
mud-cakes , below which one mating pair of C. rotundicauda resides ;
C , Mature egg emerges out of the gonopore situated at the base of the
genital operculum with a gentle pressure . 22C
 99

A B

An injured female
has been selected by
a healthy male as the
sexual partner !

Fig . 22 ( A-C)
A , Two mating pairs of T. gigas in the soft sands of Chandipur sea
beach during low tide ; B , The large female carries (undersurface) a
comparatively smaller male posteriorly on her back ; C , Shows an
amplexed pair of C. rotundicauda at Chemaguri mud flat .
Numberous juvenile Cerithedia and a small mud-skipper are found
to occur with the amplexed pair .
 Embeded F emale
Female

Satellite
Male

EXPOSED
FEMALE

C Clasping
Male

Fig . 23 ( A-C)
A , Shows an unusual amplexed pair of C. rotundicauda on the muddy
sub-creek at Chemaguri , Sagar Island ; B , Three horseshoe crabs ,
C. rotundicauda occur in a linear row - the first one is almost embedded
in the mud ; C , all the three specimens are clearly exposed just after the
removal of mud , the first one is the female while the later two are
males . The last male is being the "satellite" .
 101

Third-rail track

A B

Foot prints

Fig . 24 ( A-C)
A , Lateral view of a male T. gigas ; B , Dorsal view of the same showing
its "third-rail tracks" produced by its shell edges and tail spine , punctu-
ated by foot prints ; C , A moving pair of T. gigas ; the frontal one is
female that carries the smaller male on her back .
 102

A B
A B

C D

C D

Fig . 25 ( A-D)
A , A readily segregated female of T. gigas having dorsally situated
barnacles , moves towards the film of standing water to make a burrow
quickly ; B , Trailing activity of segregated male partner along the path of
female T. gigas ; C , Closer approximation of the male near the poste-
rior end of the female partner and burrowing instead of making further
amplexus ; D , A normal mating pair of T. gigas , engaged in making
burrow quickly into the sand (lateral view).
 103

A B

Male
claspers

C
Feast on eggs by
Corvus splendense

Fig . 26 ( A-C)
A , Shows the mechanism of clasping by the male claspers with the lateral
opisthosomatic margin of the female in C . rotundicauda ; B, Figure shows
a mating pair of C. rotundicauda partly embedded in the hard mud bottom
decorated by the faecal pellets and holes of Uca sp. Proteresia and Psueda
Mangrove herbs are also seen to be associated ; C , Shows of solitary male
of C. rotundicauda on the muddy substratum covered with Psueda sp.
 104

Fig . 27 ( A-D)
A & B , Shows the mating pair of T. gigas on exposed sandy shore with
crab holes and faecal pellets and a pair of C . rotundicauda on the
exposed mud bottom in association with juvenlie Cerithedia and smaller
Proteresia grasses . The upper furface of both pairs seems to be dried
up which indicates a long time of receding tide water ; C & D , Corvus
splendens , the predator bird sitting beside the pair and enjoying the
flesh and eggs . Sand deposition over the vacated carapace the day
after the predation is seen .
 105

E . Epifaunal associates
 106

INTRODUCTION

The carapaces , Particularly , of the adult horseshoe crabs are suitable habitats for a number
of epifaunal associates ; so much so that the American species , Limulus polyphemus has been described
as "walking museum" (allee , 1923). Most of the organisms attached to Limuli may remain attach to
several other hand surface . Thus these are considered as ectocommensals , some of which showed
symbiosis and other relationships (shuster , 1982) .
Presence of epifaunal organisms of Indian horseshoe crabs has been reported by Roonwal
(1944) . Rama Rao and Surya Rao (1972) presented a comprehensive study on the ectocommensals
infesting the two species of Indian horseshoe crabs , T. gigas and C. rotundicauda are termed them as
"mobile museums of natural history " . The present investigation elucidates a further description of the
fouling organisms in detail to relate the occurrence of epifauna with the milting phenomenon (Debnath
and Choudhury , 1991) .

Bryozoans,Anemones,Barnacles, and many others

constitute the epifaunal community on the shell of
Horseshoe crab's dorsal surface.

ONE MALE Tachypleus gigas WITH HEAVY INFESTATIONS ON THE

DORSAL SURFACE ( on both PROSOMA and OPISTHOSOMA ) OF THE
BODY
 107

OBSERVATION

A brief description of the epifaunal organisms with their taxonomic categories and size is
presented here .

1. Sea anemones : Metridium schillerianum (Stoliczka) was observed infesting heavily the dordal
surface of the body of T. gigas and C. rotundicauda . It ranged from 5 to 21 mm in its column length
(Fig. 20 A) .
2. Sessile barnacles : balanus amphitrite Darwin , measuring 2.5 to 19.8 mm in length were found
to occur profusely on the dorsal surface (rarely ventral surface) of the body of eigher species . Very
frequently goose-barnacle , Chthamalus stellatus (Poli) measuring 1.8 to 9.7 mm in length were
found to infest the carapace (Fig. 28 C) .
3. Bivalves : Several specimens of Ostrea sp. , measuring 10.5 to 28.5 mm. length were collected
from the dorsal surface of C. rotundicauda and T. gigas .
4. Gastropods : Few specimens of Cerithidea sp. measuring 8.9 to 15.7 mm. in length were
collected from both the species .
5. Amphipods : Cheiriphotis megacheles ( Giles) ranging from 4.2 to 9.2 mm. in length were
collected from the abdominal appendages , and very rarely from the empty shells of the barnacles of
T. gigas .
6. Isopods :Few specimens of Cleantis megacheles Barnard ranging from 13.0 to 15.0 mm. were
found along with amphipods from T. gigas .
7. Polychaetes : Specimens of Gattayana deludens Fauvel ranging from 14.5 to 25.0 mm . were
obtained from empty barnacle shells or from crevices of the ventral appendages . These were collected
from both T. gigas and C. rotundicauda .
8. Brachyura : A few juvenile specimens of Thalamita sp. were collected from the empty barnacle
shells present on the dorsal surface of T. gigas .
9. Bryozoa : The gymnolaemate , encrusting , amt-like bryozoan , Membranipora sp. was found on
the dorsal surface of both T. gigas and C. rotundicauda (Fig. 28C).
Beside those epifaunal associates (Rama Rao & Surya Rao , 1972) as described above , there
were also other organisms like protozoa , diatomes , algal mat particularly the colonies of Obelia sp.
But following are the two categories of ectocommensals found to occur in both T. gigas and
C. rotundicauda .
I. Turbellaria : A marine triclad , possibly belonging to the genus Ectoplana sp. were collected from
the bases of the ventral appendages and genital opercula of T. gigas and C. rotundicauda.In most cases
these were found in aggregate , mostly , on either side of cephalic swellings , i.e. below the subfrontal
area (above the ventral organ). Its tiny size ranges from 1.0 to 2.5 mm. in length . The whitish-creamy
body showed a prominent ventral sucker , a pair of cephalic eyes and distint intestinal cecae (Fig. 15 A)
(reviewed by Kawakatsu and Sekiguchi , 1989).
II. Scale - worms : Polychaete scale-worms (Fig. 15 B) , Polynoe sp. measuring 7.8 to 12.5 mm. in
length , were collected from the ventral appendages , particularly the gills and ventral grooves of both
the species of Indian horseshoe crabs . These are ectocommensals , because the similar animals were
also obtained from hermit-crabs of associate community . In aquarium they were found to swim from
horseshoe crabs to hermit crabs or hide underneath the rocks or other surfaces or vise-versa .
 Table 23 .
The % of infestations of major epifauna of Indian horseshoe crabs
(based on numberical abundance)

Sl. No. Epifauna with systematic position T. gigas C. rotundicauda

No. % No. %

1. Class : Anthozoa , Order : Actinaria 235 17.07% 149 10.60%

Metridium schillerianum

2. Class : turbellaria , Order : Seriata 130 9.44 265 18.86

Sub-order : Tricladia . Ectoplana sp.

3. Class : Gymnolaemata (Bryozoan mat). +++ - +++ -

Membranipora sp.

4. Class : Polychaeta
i) Gattyana deludens 5 0.36 10 0.71
ii) Polynoe sp. (Scale - Worms) 147 10.68 221 15.73

5. Class : Crustacea , Order : Thoracia

i) Balanus amphitrite 670 48.69 590 41.99
ii) Chthamalus stellatus 87 6.32 108 7.68
Order : Amphipoda Cheriophotis megacheles 25 1.82 - -
Order : Isopoda Cleantis natalensis 4 0.29 - -

6. Class Gastropoda
Cerithedia sp. 45 3.28 25 1.78

7. Class : Bivalvia
Order : Pseudolamellibranchiata
Ostrea sp. 19 1.38 35 2.49

8. Class : Brachyura
Thalamita sp. 9 0.65 2 0.14
108
 109
Table 24 .
The number of infested horseshoe crabs collected from 5 different stations ;
figures in parentheses indicate actual no. of animals observed .

Carcinoscorpius rotundicauda Tachypleus gigas

Male Female Male Female

Prentice Island 15(28) 8(21) 0 0

(West Bengal)

Sagar Island 51(77) 16(63) 0 0

(West Bengal)

Digha 82(131) 4(19)

(West Bengal) 0 0

Chandipur 0 0 187(333) 69(320)

(Orissa)

Dhamra 38(66) 7(50) 43(71) 11(44)

(Orissa)

% of infested animals 60.8% 23.13% 58.32% 21.93%

Head

Eye-pot

Gut

Scale-like segment

Ventral sucker
(Pharynx)

Parapodia

Intestinal divertculae

Triclad (Turbellarian) flat worm, Ectoplana sp. Polychaete scale-worm, Polynoe sp.
 110

Table 25. A&B presents the 2 X 2 contingency Chi-square tests of infested and
non-infested horseshoe crabs , C. rotundicauda and T. gigas respectively

C. rotundicauda T. gigas

0 +0 Total 0 0
+ Total

Infested 104 31 135 312 84 396

Non-Infested 67 103 170 223 299 522

Total 171 134 305 535 383 918

A.

E 0 E__0 (E__0)2 (E__0)2 / E

75.688 104 -28.311 801.512 10.589

95.3111 67 +28.311 801.512 8.409
59.311 31 +28.311 801.512 13.514 df = 1
74.688 103 -28.311 801.512 10.731

B. C. rotundicauda X2 = 43.243 P< 0.001

230.784 312 -81.215 6595.987 28.581

304.215 223 +81.215 6595.987 21.682 df = 1
165.215 84 +81.215 6595.987 39.923
217.784 299 -81.215 6595.987 30.286

T. gigas X2 = 120.472 P< 0.001

 111

Fig .A15

The ventral view of a female C. rotundicauda

showing the positions of triclad turbellarion
(A) & polychaete scale-worm (B), two
prominent ectocommensals .
B

B
 112

Egg-mass of
Tachypleus gigas

B
Massive Epifaunal Infestations on T. gigas

Morphometric measurement of Hoseshoe

crabs on sandy shore of CHANDIPUR,
Orissa.
c

Fig . 28 (A-C)

A, Enlarged view of a mass of eggs ( T. gigas ) ; B, Shows the collection

of dry carapaces for morphometric measurement at Chandipur sea
shore ; C , Shows an aged male T. gigas at Chandipur sea shore with
heavy infestations over the carapace .
 113

RESULTS AND DISCUSSION

Table 23 shows the % of epifaunal infestations based on numberical abundance of organisms

obtained from 32 T. gigas and 18 C. rotundicauda . The data clearly indicate that major infesting
animals are Balanus amphitrite , Metridium schillerianum , triclad turbellaria (Ectoplana sp) and
polychacte scale worms (Polynoe sp.) .
Allee (1923) , Andrewa (1942) , Gorham (1904) , Humm (1942) , Pearse (1947 , 1949) ,
Shuster (1955 , 1958) , Verrill (1893 , 1895) , Wheeler (1894) have reported a variety of epifaunal
organisms from L. polphemus . But Rama Rao & Surya Rao (1972) are the first persons to report these
from Indian horseshoe crabs . In their paper (Opt. Cit.) they tried to find out an inter-relationship
between the infesting animals and the horseshoe crabs . The present investigation does not agree with
the statement - "the amphipods , isopods and polychaetes are probably attracted for eating the egg mass
clustered around the abdominal appendages of King crabs" (Rama Rao & Surya Rao , 1972) , since the
evidence of possessing clustered eggs in the ventral appendages of the female 'crabs' , has already been
discarded (Debnath , 1985 ; Iwanof , 1933 ; Roonwal , 1944 ; Sekiguchi et. al. , 1977) .
The number of horseshoe crabs infested with epifaunal organisms in given in the table 24
which clearly indicates that male horseshoe crabs are being infested with ectocommensals (58.92%)
more than that of the female crabs (22.24%) . Out of 305 ( 171 M + 134 F ) C. rotundicauda observed
only 31 female (F) showed epifauna with less or sometimes moderate infestations . Heavy infestations in
females were very rare . On the other hand , amongst 918 ( 535 M + 383 F ) T. gigas only 84 females
(F) showed epifaunal infestations . But for male (M) C. rotundicauda and T. gigas it accounts 60.81%
and 58.31% infestations respectively .
The Chi-square test of the infested to non-infested horseshoe crabs reveals that in both
cases males are highly vulnerable to sustained growth of epifaunal organisms on their carapaces than
the females (for C. rotundicauda , X2 = 43.243 , P < 0.001 and for T. gigas X2 = 120.471 , P< 0.001)
(Fig. 28C)
The high frequency of epifaunal infestations in males than in the females may be explained
in two ways :

i) Since mating amplexus takse place in the shallow seas before the commencement of spring and
summer months (debnath , 1985 ; Debnath & Choudhury , 1988a) the males remaining attached to the
dorsal carapace of the female get exposed for easy access to the epifaunal organisms . This hypothesis
can be approved by the findings of fouling organisms mostly on the solitary females , very rarely on the
mated females .
ii) The more appreciable second assumption is that males do not molt further after attaining
sexual maturity (Shuster , 1955;58) and thereby this shells become suitable habitat for epifaunal growth ;
while as the females molt more frequently , at least onece in a year (shuster , 1950 , 1954) ; its shells are
generally remain free of fouling organisms and it seemed likely that the larger size attained by females is
due to successive moltings (Rudloe , 1980) .
The frequency of molting is probably a deterent to sustained attachment of ectocommensals .
Thus the first attachment is noticeable on Limuli larvae and is most prevalent on adult Limuli, because
they do not molt as frequently if at all , paricularly the males , permitting continued individual growth as
well as population development of the attached animals (Shuster , 1955/58) . This is further confirmed
by the fact that all immature and most young (adult) Limuli are found free of "encrusting" organisms
(Shuster , 1957) .
 114

Although distinction between ectocommensalism and parasitism has not made , the marine triclad Bdelloura
has been categorised as ectoparasite of Limulus (Ryder , 1982 ; Verrill , 1893) and Ectoplana for T.
gigas and C. rotundicauda (Sluys , 1984). Carapaces of adult horseshoe crabs are found frequently with
encrusted epifaunal invertebrates , particularly barnacles , blue mussels and slipper limpets which could
potentially be useful in approximating ages of adult horseshoe crabs (with Crepidula fornicata , a gastro-
pod ectocommensal cf. Botton & Ropes , 1988) . In addition to providing insights into the age-structure
of an adult horseshoe crab population , studies of epifauna may be useful in determining whether discrete
spawning populations throughout a coastal range exist or not .
In fine , horseshoe crabs provide a better shelter to a variety of epifaunal associates i) for their
transport from one locality to the other (since the host 'crab' has effective migrating habit) as in the case
of transportation of thousands of oyster-drills on the carapace of single Limulus (Mackenzie , 1962) ,
ii) for facilitating feeding of barnacles and other sessile forms , and iii) for chemouflaging itself from
the predators . According to Shuster (1957) infesting organisms provide a clue to the identification of
horseshoe crabs of different geographical localities and "encrustations" , if identified , on fossil Limulidae
may give information on the ecology and growth stage of the fossil .
 115

PART - II
STUDIES ON FEEDING BEHAVIOUR ; FOOD ;
FEEDING AND DIGESTION

................ it is to be noted that

the functional activities of the
neuroendocrine system of
horseshoe crabs are , probably
attributed with the moulting
phenomena , visual system and
mating behaviour.
 116

INTRODUCTION

From several recent reviews of the natural history of horseshoe crabs (Shuster ,1979 ,1982 ;
Rudloe , 1979a , 1981 ; Sekiguchi and Nakamura , 1979 ; Sekiguchi , 1988) , it is evident that relatively
very little is known of the merostomate's feeding biology , inspite of its familiarity to a large number
of zoologists and palaeontologists , and its growing importance in biomedical research (Cohen et. al. ,
1979 ; Pearson and Weary , 1080 ; Nobitsky , 1982 , 1984) . Information on the feeding biology of
horseshoe crabs is limited (lockwood , 1870 ; Fowler , 1908 ; Shuster , 1950 ; Smith and Chin , 1951 ;
Smith , 1953 ; Smith et. al. , 1955 ; Botton , 1981) . The first comprehensive study of feeding
behaviour of Limulus polyphemus is of Botton (1984 a, b) which revealed a resonable method-
ology to conduct the gut-content analysis in Indian horseshoe crab , Tachypleus gigas (Debnath ,
1985 ; Debnath and Choudhury 1987) . Botton and Haskin (1984) provided a more intensive study of
the food habits of L. polyphemus recovered from the continenta l shelf of Delaware Bay and the
subsequent analysis of their stomach-contents .
Except Schlottke (1934a, 1934b, 1935) ; Lockhead (1950) and Tanimoto and Kondo
(1982) , so far no attempt has been made to study the digestive physiology of Merostomates . It is
for the first time that the gut contents of T. gigas have been examined in context of the digestive
enzymes secreted by the organism ( Debnath et. al., 1989). This part of the thesis discusses in brief the
digestive system , feeding and food-items in context to digestive physiology and finally deals with
the trophic level to ascertain the possible position of the Indian horseshoe crabs in coastal or
estuarine trophic level .

Corvus predator
Deliceous dish Carnivore predator

Large predator fish Small goboid fish

?
Crab's eggs
OR SED C
SH on
N H BA PHI

OE
DIA as TRO

SE

Tachypleus Shrimp Carcinoscorpius

f IN WEB OF
2B
AB UDIE FOOD ORM
.3
F
FIG

D
TH ELS IFIE

So

Polychaetes Gastropods Copepods Bivalves Ophiuroids

&
L
LE IMP

T
S

ES
S
V
A

CR

Plants & Detritus (organic decompose)

 117

OBSERVATION

DIGESTIVE SYSTEM

The early works of merostomate digestive system were on L. polyphemus ( Patten and
Redenbough , 1899 ; Schlottke , 1935 ; Shuster , 1948) , but not on that of the Asian horseshoe crabs ,
except for a short description in T. tridentatus ( Shoji , 1929) .
The digestive system of horseshoe crabs consists of two main parts ; the digestive tract and the
mid-gut gland . The digestive tract is a J-shaped simple duct sub-divided into 3 parts : i) the fore-gut ,
comprising mouth , oesophagus , and proventriculus ; ii) the mid-gut or stomach-intestine ; and iii) the
hind-gut , comprising the rectum and anus .
The mid-gut gland , filling most of the prosoma , consists of two different tissues ; the hepatic
diverticulae i.e. the blind branches of the hepatic ducts arising from the intestine , and the yellow connec-
tive tissue surrounding the mid-gut diverticulae . In adult horseshoe crabs , the mid-gut diverticulae are
located only in the prosoma , but the yellow connective tissue is seen in the opisthosoma , on either side
of the intestine .

1. Digestive tract :

The mouth is a longitudinal slit that opens on the ventro-median surface of the prosoma sur-
rounded with three kinds of mouth parts : a pair of chelicerae , coxae or gnathobases of five pairs of
walking legs , and a pair of chilaria . The elastic peristomal cuticle enables the mouth parts to move freely
The oesophagus , the wall of which is provided with developed muscles , passes upwards from the
mouth , through the vascular ring within which the circum-oesophageal nerve ring is situated , in fornt of
the endosternite it then runs forwards to the more swollen proventriculus . It is often regarded as the
"stomach" or "gizzard" ; however , it is not a true stomach of endodermal orgin , rather it belongs to the
ectodermal foregut and called as the crop . At the half-way point , it is strongly bent backwards , divided
into front and hind parts . The wall of the proventriculus , i.e. the crop (front) or gizzard (hind) , is
provided with well-developed muscles , is the thickest one in the whole digestive tract . Externally , a
dorsal notch is often seen that marks the distinction between crop and gizzard ( Fig. 32 C) .
The posterior end of the proventriculus (hind-part = gizzard) narrows sharply into apyloric
valve , protruding into the intestine . The first part of the intestine thus has a swollen appearence that
covers the pyloric valve and regarded as the true stomach . A marked constriction indicates the boundary
between the gizzard and the stomach-intestine . The intestine is a soft and wide tube stretched between
the prosoma and opisthosoma , lying longitudinally under the heart and running to near the posterior end
of the opisthosoma . A short rectum leads to the anus , opening ventrally at the base of the telson which is
guarded with a pair of anal valves . There is no boundary such as valve or constriction between the
intestine and the rectum . The wall of the rectum , with its well developed muscles , is thicker than that of
the intestine . One on each side of the anterior part of intestine lies the hepatopancreas ; paired ducts from
each of them enter into the intestine. Openings of the first pair lie slightly antero-ventral and the second
one slightly postero-dorsal (Fig. 32C) .
The length and diameter of five major parts of the alimentary tracts (10 mated pairs of T.
gigas ) are given in the following table as mean values expressed in mm. and it was also found that the
total mean length of male tract as 110.38 mm (range : 105.0 - 119.4 mm) ; while it was 153.80 mm
(range : 147.5 - 163.5 mm) in female .
 118

Length and diameter of different parts of the digestive system

MALE FEMALE
(mm) (mm)
Length Diameter Length Diameter

1.Oesophagus 28.75 3.52 34.50 8.12

+5.97 + 0.56 ±4.08 + 2.03

2.Proventriculus (Crop) 14.53 8.75 21.76 12.45

+ 2.19 + 1.87 ±1.25 + 3.16

3.Gizzard 16.98 7.42 20.44 7.75

+ 1.83 + 1.34 ±1.40 + 1.13

4.Intestine 39.45 4.52 50.25 8.87

+ 2.41 + 0.81 ± 2.89 + 2.29

5.Rectum 25.25 7.76 37.12 11.05

+2.25 + .047 ± 3.42 + 5.24

The inner surface of the oesophagus is deeply folded into 6-7 longitudinal ridges and furrows ,
which are covered with a transparent moltable cuticle . In fore part of the proventriculus (crop) the inner
surface has many longitudinal ridges as extensions of those present in the oesophagus . But in contrast ,
the hind part i.e. , gizzard has 13-15 , usually 14 , deep longitudinal ridges and furrows , the ridges are
remarkably deeply notched into several chitinous denticles; the hardened cuticle of which often coloured
blackish brown . These features show that this part is undoubtedly a place for mechanical manipulation or
a masticatory stomach (Yamasaki et. al. , 1988) . In the pyloric valve , the toothed lines are reduced into
6 fine serrated ridges . This valve opens into the stomach or the first swollen part of the intestine with 6
papillae (Fig. 32 C) .
The oesophagus , the proventriculus , the gizzard including the protruding pyloric valve into
the stomach-intestine , all are characteristics in having the muscular wall on the outer part and transpar-
ent or thick moltable cuticle in the inner surface . These common features show that the parts have the
same ecotodermal or the stomodeal orgin (Yamasaki , et. al. 1988) .
The inner surface of the intestine is brownish in colour , has a thin mucous layer , instead of
the cuticle . The excreted faeces of the horseshoe crab are surronded by a mucous envelope . One or
more such faecal blocks or threads enveloped with fine mucous layer were observed . These features of
the intestine , which are quite different from those in the oesophagus and to the proventriculus , show that
the intestine is of endodermal orgin (Yamasaki et. al. , 1988) .
In the final phase the inner surface of the rectum is again folded into 6 or 7 longitudinal furrows
and ridges and covered with a transparent moltable cuticle , which further confirms the rectum originating
 119

2. The mid-gut gland :

According to Yamasaki et.al. ( 1988 ), the mid-gut diverticulum is a blind branch of the hepatic
duct , 100 - 200 µm in diameter . These are distributed throughout the prosoma surrounded with yellow
connective tissue in an anastomosis with the midgut gland and the gonad . The wall of the midgut diverticu-
lum consists of two layers - the inner hepatic epithelium and the outer muscle layer . The former is
columnar cells and a small number of conical cells . The former cell types include several large secretory
granules , the distal part of which is often cut off from the cells and floating in the lumen of the diverticu-
lum. The conical epithelial cells lie sporadically among the columnar epithelial cells . In form and stainibility
the granules in the conical cells are similar to the granules stored in the yellow connective tissue . A
remarkable level of non-specific esterase enzyme activity has been noted in these granules of the conical
cells (Tanimoto and Kondo , 1982) . Fage (1949) described the midgut diverticula as partly located in the
opisthosoma in L. polyphemus . In young L. polyphemus of the 1st and 2nd instars , some branches of
the hind pair of hepatic ducts enter into the opisthosoma (Shuster , 1948) , but the mid-gut deverticulae
are located only in the prosoma in adults of any species .

3. Yellow connective tissue :

Yamasaki et. al. (1988) described this specialized tissue in great detail . It is a soft mesen-
chyme-like tissue , yellow or light brown in colour ; consists of a single type of large yellowish cells ,
which are often filled with many eosinophilic granules . Nutritive material (oesinophilic granules) may be
stored in this tissue and released into the blood. This tissue on the other hand , may be homologous with
the chloragogen tissue of certain annelids , because of its colour , its celluar constituents , and its location
around the intestine . Most of the yellow connective tissue composes the midgut gland together with the
midgut diverticula . For this , several authors described the opisthosomatic midgut gland as the
hepatopancreas or liver in adult crabs (e.g. , Fage , 1949 ; Shoji , 1929) .
The digestive system of the four species of horseshoe crabs have a good many morphological
similarities (except a few) in minute details , such as longitudinal ridges in the inner surface of the
oesophagus , the proventriculus , and the rectum . These similarities may be due to evolutionary
conservation , rather than convergence , and the basic feeding behaviour remains same and unchanged in
all extinct and extant horseshoe crabs , which leads to the final conclusion that the horseshoe crab's
digestive system retains some primitive features like the well-defined yellow connective tissue
(Manton,1977; Yamasaki et. al. , 1988) .

GUT - CONTENT ANALYSIS

Major food-items of T. gigas occurring at Digha sea shore , were comprised of vascular plant
parts , foraminiferan shells , cnidarians , polychaetes , tabanid larvae , crustaceans , specially amphipods
and junvenile shripms , soft-shelled molluscs (both bivalves and gastropods) , ophiuroids , and goboid
fishes . The major food-items recovered from the gut-contents may grossly be grouped as below :
 120

Male Female

1. Polychaetes : 11.46% 16.28%

2. Gastropods : 11.64% 16.94%
3. Bivalves : 19.04% 16.28%
4. Crustaceans : 6.35% 8.47%
5. Tabanid larvae : 35.62% 18.89%
6. Ophiuroids : 5.46% 16.94%
7. Miscellaneous : 10.40% 6.18%

The miscellaneous category includes unidentified prey items of various taxa and few
zooplanktons besides few goboid fishes and synthetic fibres . Several varieties of foraminiferans ,
Casuarina needles , vascular plant debris and fruits mixed with sand-particles and pebbles were also
found in the guts (Fig. 29 , 30 & 31) .
Now , a detailed description of food items recovered and identified upto possible lowest taxa
can be presented :

Bivalve molluscs :
Several kinds of bivalve shells (intact very rarely) were found . Usually fragments of the bivalve
shells made it difficult to identify the prey-item. In most cases shells were found in complete corrossive
form , although remained intact . But juvenile bivalves less than 0.5 mm in length were frequent in the gut
contents as indicated by unbroken shells . The possible taxonomically identified catgories are : Anadara
sp. , Dosinia sp. , Placenta sp. , Macoma sp. , Solen sp. , Neosolen sp. and Teredo sp. etc. (Fig. 30B).

Gastropod molluscs :
These were also very difficult to identify , because most of the shells became corroded ,
crashed or fragmented . However few of them were identified as Cerithedia sp. , Littorina sp. ,
Assimenea sp. , Nerita sp. , Cassidula and Cymia sp.etc. (Fig. 30C) .

Annelids :
Polyehaetes were the only annelid worms recovered with intact head or tail and sometimes
complete bodies of juvenile forms . The major polychaete worms are Gattyana sp. , Phyllodoce sp. ,
Nereis sp. , Perinereis sp. , Glycera sp. , Sabellaria sp. , Polydora sp. etc. (Fig. 31A) .

Arthropods :
Amongst this category the major food items consist of crustaceans , mostly unidentified .
These were mysid shrimps , gammarid amphipods , copepods (e.g. Microstella sp. ) , decapods (e.g.
Crangon sp.) and Cirripeds (Fig. 31B) .
Under Insecta , the most dominating food item is a tabanid larva , identified as Atylotus
agrestis . The other rare insect larva was Chironomus sp. Frequently body parts , usually , appendages of
unidentified insect were also recovered from the gut (Fig. 29C) .

Miscellaneous food matters :

These include numerous foram shells like Ammonia beccarii , A. tepida , Asterorotalia
dentata , A . multispinosa , A . trispinosa , Bolivina sp. , Elphidium sp. , Ouinquloculina sp. These
benthic foraminiferans are found along the south-east coast of Bay of Bengal (Mazumder , 1992, pers.
 121

communication ) . Other miscellaneous food matters include unidentified bryozoans , nemarteans ,

turbellarian trematodes , few nematodes , eggs , synthetic fibres and calcareous spicules of Echinoidea ,
siliceous crystals etc. mixed with sand particles . (Fig. 29A , 30A) .

Vascular plant matters :

Vascular plant parts include Casuarina needles , plant fruits , algal bodies etc. These plant
matters constitute a major part of the gut-content (Fig. 29B)

Teleost fishes :
Three goboid fishes with distinct ventral suckers (?) , fins with fin rays and scales were found
from the gut-content of T. gigas collected from Chandipur sea shore (Fig. 29 C(A) .
Remark - I : Razor clams (Solen sp.) brachyuran crabs ; and polychaetes etc. were the major food
constituents of C. rotundicauda collected from Prentice Island (West Bengal , Sundarbans) and estua-
rine mangroves of Dhamra (Orissa mangroves) .
Remark - II : Beside the above food items , T. gigas of Digha sea shore possessed several juvenile
anthozoan actinarians (unidentified) .
Remark - III : Fragmented shells , spicules , sands and other solids were found in compact ball-forms
within the proventriculus , just below the dorsal-notch which marks the junction of crop and gizzard ,
ready to be ejected out through the mouth . Such "shell-balls" were found in 15 specimen-guts out of 72
dissected for content-analysis (Fig. 32C) .
Table 26 shows the measurement of food-items recovered from the gut-contents of T. gigas .
This table clearly shows the length and breadth (both expressed in mm) of food matters . The maximun
body fragment of polychaete worm measured 25.8 mm and it was 30.0 to 40.0 for goboid fishes ; while
the minimum length ranges from 0.3 to 0.5 mm for juvenile molluscs . Most of the junenile molluscs
were observed in the semidigested body of polychaete worms and tabanid larvae ; few from both gizzard
as well as rectal content of horseshoe crabs . Similarly foram shells were obtained from both gizzard and
rectal content which also measured less than 1 mm. in diameter .
In Table 27 the frequency ( f ) and number of food items along with their ratio ( r = n/f ) have
been presented . The data are arranged in three major columns , that is , for 2 solitary males , for 15 mated
males and 15 mated females . The 'r' values clearly indicate that the solitary males consumed more
food than the mated males except in the cases of miscellaneous category . At the same time , it is found
that mated females had consumed food much more than either the solitary or the mated males . In terms
of percent if 'r' values compared the following assumptions can be drawn :

i) Adult males (both solitary and mated combained) consumed only 65.78% food than those of
females .
ii) Mated males alone can consume as much as 92.65% food if compared with the 100% of solitary
males , i.e. 7.35% less .
iii) 61.65% consumpsion by mated males and 66.65% consumpsion by solitary males , if compared
separately with that of the mated females .
Table 28 presents the weighed values of gut-contents of 15 mated males and females , respec-
tively .This observation apparently reveals the high gut-content value in females (0.952 + 0.47) than the
males (0.862 + 0.80) irrespective of their body weight .
 122

DIGESTIVE PHYSIOLOGY
The results of the examination of the gut content of 20 specimens, 10 of each sex , are
recorded in Table 29 in relation to their body weights and inter - ocular length (IOL in mm.) . Abundant
food matters were found in seven females out of ten , whereas only one male contained abundant food .
Vascular plant debris were found in all females and in all males but one . Numerrical abundance showed
that major food items are molluscs , polychaetes and crustaceans (mostly gammarid amphipods) . Numer-
ous foraminiferan shells mixed with sand particles were found in the gut-contents (Table 29) .
The results of digestive enzyme assay have been presented in table 30 . The activity of acid
protease is highest in gizzard and lowest in oesophagus , whereas alkaline protease is highest in intestine
and lowest in gizzard . Esterase shows maximun activity in oesophagus , whereas in the other three regions
studied , the activity is reduced by 50% . Cellulase activity is highest in gizzard and oesophagus , but for
invertase and amylase , highest activities are found in digestive gland and lowest in the oesophagus .

TROPHIC LEVEL :
A simplified food web based on Shuster (1982) , comprising various trophic levels , has been
constructed (Fig. 32B) after analysing the gut contents and predatory phenomena of horseshoe crabs . In
this study it has been observed that horseshoe crabs preyed on various prey forms from micro-to
macro-organisms . For instance , an important dietary components of horseshoe crabs was polychaetes
which were dependent on juvenile gastropods and bivalves . Similarly larger soft shelled molluscs are
detritus feeders . On the other hand , no ophiuroids are used as food by man , and none are venomous ; but
they have a considerable indirect economic importance in view of their immense number and consequent
significance in natural food chains involving commercially sought species like horseshoe crabs , T.
gigas . Most of the pelagic forms of polychaetes and zooplanktons like calanoids ( a copepode) , mysid
shrimps , and Microsetella sp. etc. are common food items of the mated males of horseshoe crabs ,
as they did not come access to the benthic communities , were dependant on the pelagie micro-
organisma , while in motion along the back of the carrier female in amplexus . The goboid teleosts ,
on the other hand , are either micro-carnivores or detritivores .
In the above category horseshoe crabs can be regarded as a predator . While , the crow , Corvus
splendens was found to predate on horseshoe crab's egg. Thus a very complex food chain in the form of
a net-work (food web) is being obtained . In the present investigation , it is evident that horseshoe crabs
might be regarded as primary , secondary tertiary consumer as and when the circumstance demands
certain trophic level for any individual nutritional flow system . Higher trophic level like crows , jackles
or even human-being ( in Thailand, coastal China including Hong Kong as referred by Sekiguchi and Nakamura , 1979; & also
a pers.comm. with Huang Qin, Helen Chiu and Brian Morton, 2003....."the fact that they are, locally and elsewhere in China, fished,
sold in markets and eaten as a specialist sea food." ) seem to be under the tertiary consumer category as observed in
the present investigation .

FEEDING
The mechanism of feeding in horseshoe crabs have been described by a number of authors ,
begining with Lockwood (1870) . Usually small macrofauna and juvenile forms are initially captured by
the animal using the chelate walking legs . These ventral appendages along with chelicerae , chilaria and
genital operculum , manipulate the food-items to the gnathobases of the legs . The mouth lies between the
1st pair of legs . Posteriorly from it , and between the dentate process of the coxae which form the
gnathobases , runs a groove called the food basin ( Fig. 15; * it may be comparable with the pro-meso-
sternite as suggested by the expert, Prof.C.N.Shuster,Jr.1993) . The chitinous gnathobases are fortified
 123

Table 26

Measurements of various food items (all expressed in mm) of T. gigas

Taxonomic category Items Length (mm) Breadth (mm)

Annelida
Polychaete jaw 0.9 - 1.2 0.2 - 0.3
Polychaete body(fragmented) 2.0 - 25.8 1.8 - 2.2
Polychaete setae 1.3 - 1.6 -
Polychaete head (complete) 3.7 - 5.4 1.6 - 2.1
Juvenile Polychaete (complete) 1.8 - 2.0 0.3 - 0.4
Mollusca

Bivalve shell (complete) 5.0 - 8.0 2.5 - 4.3

Bivalve shell (fragmented) 3.5 - 6.2 -
Juvenile bivalve 0.3 - 0.5 0.2 - 0.3
Gastropod shell (complete) 4.0 - 7.2 3.1 - 4.3
Gastropod shell (crashed) 3.2 - 5.8 2.8 - 5.0
Gastropod shell (fragmented) 2.3 - 2.5 -
Crustacea
Amphipod body (complete) 2.7 - 4.0 0.8 - 1.3
Crustacean head(semidigested) 3.5 - 4.0 0.7 - 1.5
Crustacean appendages 1.0 - 4.2 -
Zooplankton (complete) 0.4 - 0.7 -

Insecta
Tabanid larvae(Atylotus 4.5 - 5.0 0.2 - 0.3
agrestis)

Cnidaria
Juvenile anemones (?) 3.0 - 6.8 0.5 - 0.7
Hydroid colony 3.8 - 4.2 -

Foraminifera
( shells of various kinds) - 0.3 - 0.7

Pisces Goboid teleosts 30.0 - 40.0 5.7 - 10.5

Eggs Unidentified eggs - 0.1 - 0.7

with sharp spines to grind molluscan shells and other prey (Shuster , 1982) . Anteriorly along the food
basin , food is passed to the mouth while being triturated , and then enters a muscular , chitinous gizzard
or proventriculus , where additional grinding occurs . Food is grounded in the gizzard and admitted to the
stomach-intestine through the pyloric valve . Coarse fragments are ejected through the mouth
( Lockhead , 1950) as bolus forms , found in the present investigation , at least in 15 guts dissected out .
 124

Table 27 .

Occurrence of food items in digestive tracts of 72 Tachypleus gigas collected from Digha
sea shore , West Bengal , India ; during the summer of 1985 ( f = frequency of food items occurred
at least one in the gut-content analysed ; n = total number of food items counted )

Taxonomic
category 42 solitary males 15 mated males 15 mated females
of food items 36 with food 7 with food 15 with food

f n r f n r f n r

Arthropoda
Crustacea 18 29 1.611 5 7 1.400 10 26 2.600
Insecta 29 63 2.172 6 10 1.666 15 35 2.333

Annelida
Polychaete 28 35 1.250 4 4 1.000 13 42 3.230

Echinodermata
Ophuroidea 11 17 1.545 2 3 1.500 10 15 1.500

Mollusca
Bivalvia 20 75 3.750 4 12 3.000 12 40 4.166
Gastropoda 16 36 2.250 3 7 2.333 11 33 3.000

Miscellaneous
Unidentified 28 31 1.107 7 12 1.714 15 28 1.866
and others

Vascular plant 28 22 + 3 2+ 15 2+
matters 5 ++ __ 1 ++ __ 7 ++ __
1 +++ 0 +++ 6 +++

*r = ratio of n / f ; + , present ; ++ , moderate ; +++ , high ; and

Foraminiferan shells were excluded from the count .
 125

Table 28 .
Weight of gut-contents of 15 segregated males and females (expressed in
gm.) UG , EG & GC , weight of undissected gut , emptied gut and gut-
contents ; FP , SP & DP , food , sand and debrices present .

UG EG GC FP SP DP Body
weight

3.75 3.24 0.51 + ++ - 130.0

4.28 3.87 0.41 ++ + + 155.0
5.03 4.25 0.78 ++ ++ + 120.0
3.43 3.03 0.40 + + ++ 175.0
MALE 2.95 2.85 0.10 - + + 200.0
N=15 2.57 2.55 0.02 - + - 135.0
4.87 3.75 1.12 - + + 180.0
5.25 3.96 1.29 ++ +++ + 140.0
5.04 3.87 1.17 ++ ++ ++ 210.0
4.85 3.25 1.60 + +++ + 125.0
5.86 3.28 2.58 ++ +++ ++ 120.0
6.23 3.95 2.28 + +++ ++ 170.0
4.28 3.57 0.71 + ++ +++ 155.0
3.05 3.05 - - - - 140.0
2.57 2.56 0.01 - + - 135.0

4.87 4.53 0.34 ++ - - 405.0

5.95 5.45 0.50 +++ + ++ 490.0
6.35 5.03 1.32 +++ ++ ++ 510.0
6.54 5.15 1.39 +++ ++ + 425.0
5.85 5.54 0.31 ++ - - 485.0
FEMALE 6.83 5.59 1.24 + +++ - 515.0
N=15 6.25 4.83 1.42 ++ +++ - 355.0
5.56 4.71 0.85 ++ + ++ 505.0
4.95 4.50 0.45 ++ + - 480.0
4.87 3.92 0.95 ++ ++ ++ 545.0
5.08 3.56 1.52 +++ + +++ 490.0
6.05 4.47 1.58 +++ + +++ 505.0
5.34 4.09 1.25 ++ +++ ++ 410.0
4.87 4.55 0.32 + - ++ 405.0
6.35 4.50 0.85 +++ + + 540.0
 Table 29 .
Data on gut contents of 20 mated Males and Females of Tachypleus gigas .

Sex Weight (g) IOL(mm) Gizzard Plant Foraminiferan Ophiuroids Mollusc Polychetes Crustaceans
content debris shells (Echinodermata) Shells

Male
N=10 130.0 73.0 + ++ - - - - -
140.0 76.0 + + - - - - -
120.0 71.0 ++ ++ 8 1 22 - 1
155.0 74.0 ++ + 150 1 4 2 1
101.0 75.0 + + - - - 1 -
120.0 69.0 + + 7 - - 4 1
150.9 73.0 ++ + - - 3 3 1
120.0 71.0 +++ + 4 - 2 3 1
195.0 78.0 ++ + - - 5 1 -
125.0 71.0 - - - - - - -

------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

Female 405.0 92.0 + + 5 1 2 - -

N=10 409.0 101.0 ++ +++ 150 - - 1 -
510.0 103.0 +++ +++ 100 15 8 3 1
425.0 92.0 +++ ++ 75 1 6 2 1
485.0 101.0 +++ ++ 15 - 4 4 -
515.0 105.0 + ++ 8 1 - 3 2
355.0 89.0 +++ ++ 2 3 11 1 2
505.0 102.0 +++ ++ 12 - 7 - 1
480.0 103.0 +++ +++ 3 19 15 4 1
585.0 107.0 +++ +++ 5 2 12 2 1

IOL , Interocular length . + , present ; ++ moderate ; +++ , high

126
 127
Table 30

Dig esti
Digesti ve enzyme assa
estiv assay y of T. gigas
gigas
(Mean + SD values are presented )
1 2 3 4
Enzymes Oesophagus Gizzard Intestine Hepatopancreas
Assayed n=20 n=20 n=20 n=20

Acid Protease 0.192 + 0.015 0.679 + 0.061 0.464 + 0.037 0.259 + 0.023
(mg. tyrosine)

Alkali Protease 0.406 + 0.053 0.372 + 0.059 0.756 + 0.113 0.642 + 0.098
(mg. tyrosine)

Esterase 0.500 + 0.055 0.260 + 0.031 0.273 + 0.035 0.296 + 0.034

(mg. B-napthol)

Cellulase 10.944 + 1.061 11.529 + 1.037 7.736 + 0.618 9.351 + 0.912

(mg. glucose)

Invertase 3.462 + 1.290 4.683 + 0.601 5.899 + 0.612 6.059 + 0.557

(mg. glucose)

Amylase 13.587 + 1.290 15.640 + 1.801 15.658 + 1.733 16.608 + 2.011

(mg. glucose)

MO
UT
H
US
AG
PH
SO HEPATOPANCREAS
OE
OP
CR

T
U
-G
ID
M
A HIND-GUT
RECTUM

ANUS

INTESTINE
GIZ STOMACH
ZAR
D
L
NA

FOOD BOLLUS
RID ITUDI
S
GE
NG
CON

B
LO
ICA

D
LVA
LVE

H LAR
TE TICU
ET
CU

C
Fig. 32 C , Shows different parts of the gastro-intestinal tract of horseshoe crab .
 128

B C

Fig . 29 (A-C)

A, Shows various kinds of foram shells obtained from the stomach content ; B , Plant
parts comprising fruits , vascular structures , Casuarina needle and others ; C ,
spicules , polychaete jaw , body of tabanid larva , segments of ophiuroid arm and full
goboid fish body have been recovered from the stomach content of T. gigas .
 A

Fig . 30 (A-C)

A, Various parts of food matters , the most important being the razor clams (RS) occur in the gut contents
of C. rotundicauda ; B , Different bivalve prey shells of both fragments and intact juveniles occurring in
the gut ; C , Shows the broken as well as intact gastropod shells in the gut of T. gigas .
 130

Fig . 31 (A&B)
A, Shows pelagic forms of polycheates and intact body parts , particu-
larly the head and tail regions ; B , Plate showing the intact as well as
broken or partly corroded crustacean body parts . The most conspicu-
ous is being the gammarid amphipod (C) .
 131

A B

C
Fig . 32 (A-C)
A, Dissection of the alimentary tract for its primary preservation in 3.7% formalin .
Numerous mature eggs are also seen in the tray recovered from the female's
prosoma ; B , A simplified trophic structure showing its various levels of organization
for both T. gigas (Tg) and C. rotundicauda (Cr) . Polychaetes (P) , gastropods (G)
, mysid shrimps (M) , bivaves (B) and ophiuroids (O) that depend primary on the
benthic detritus and plant life ; C , Shows various parts of the gastro-intestinal tract of
horseshoe crab .
 132

DISCUSSION
Molluscs are the most importent food item in terms of numerical abundance to adult
T. gigas and L. polyphemus of Delaware Bay (Botton 1984a,b) during the season .Next to its stand the
numerical abundance of arthropods e.g. gammarid amphipods and tabanid larvae . On the continental shelf
of New Jersey , crabs preyed on a wide variety of benthic organisms , primarily molluscs , arthropods and
annelids (Botton and Haskin , 1984) . The size inequality between adult male and female horseshoe crabs
generated the hypothesis that diets of males and females might differ . Females are larger than males ,
which might enable females to feed on large prey . Futhermore females can dig deeper than males and
might therefore have access to such type of fauna which are not available to males . However , based on
gut contents , males and females in the present study show similar diets and food preferences . In the
aquarium experiment , Botton (1984b) observed similar response by the males and females ,
furthermore , males were to feed more .
The above results of L. polyphemus do not corroborate with the results obtained in the present
investigation . Adult mated females of T. gigas were found to consume more food than either the solitary
or mated partner males . On the other hand , solitary males are found to consume more food than the
mated males . These findings lead to the dorsal opisthosoma of the female partner for a long time they do
not have access to the bottom benthic community ; while this was more easy for the carrier females . At
the same time solitary males come in direct contract to the benthic community for feeding . This hypoth-
esis is further confirmed by the presence of all zooplanktons or pelagic prey found in the gut-content of
mated males .
The tenable alternative hypothesis (Botton , 1984b) explaining the food selection in horseshoe
crabs involves chemically mediated preferences . Both gnathobase spines and chelae of the walking legs
possess chemorecaptors (Barber , 1956 ; Wyse , 1971) which are supposed to be involved in food selec-
tion .
As regards the preferential consumption of prey materials of L. polyphemus , Botton's (1984
a,b) analysis shows that the 'crab' lays its preferential importance on the large over small molluscan shells
and soft shells over the thicker one in both the sexes . Of course this type of observation in the gut
contents of T. gigas and C. rotundicauda could not be established , at least in this geographical locale.
Effects of predator's differential prey digestibility may induce bias results in the identification
and enumeration of ingested prey materials in favour of taxa with shells and exo-skeletons (Botton 1981)
. Keeping this in mind , the gut contents were meticulously examined , and molluscs were found to be the
most abundant animal prey followed by the arthropods and polychaetes (Botton , 1984b) . Consumption
of plant detritus of T. gigas is supported by the report of ( Botton , 1984a,b) i.e. 90% L. polyphemus
consumed heavy amount of vascular plant matters , although the nutritional value of plant to this predator
is not known .
In the present investigation , gut content was seen to reflect similar food preferences between
the two sexes of the species studied , through the females were found to have more ingested matter in gut
than the males . This may be due to the fact that in the coupled T. gigas , the smaller males clings to the
back of the larger female , which has its chelae free for foraging . These observations led the author to
infer that T. gigas which grazed along the sea shore , was a dietary generalist (Debnath and
Choudhury , 1987) . This observation has also been endorsed by Botton and Haskin (1984) . The
presence of proteases , esterases and carbohydrases in the digestive tract and associated gland lends
further evidence to the above assumptions (Debnath et. al. , 1989) .
Lockhead (1950) reported in Limulus ( = Xiphosura) polyphemus , that digesion occurred in
the stomach-intestine by means of enzymes from hepatopancreas (midgut gland) , except for splitting
dipeptides , which occurred (Schlottke , 1934a,b) . A second set of enzymes active in acid medium in
 133

the foregut has been reported which may be the reason why acid protease activity is highest here . But the
stomach - intestine which received secretions from hepatopancreas has high alkaline protease activity
(Lockhead , 1950) . Among proteases , the alkaline and among carbohydrases , amylase are the most
abundant . A remarkable amount of non-specific esterase has been reported in T. tridentatus by Tanimoto
and Kondo (1982) in the conical cells of the hepatopancreas , but how this esterase has occurred abun-
dantly in the oesophagus of T. gigas is yet to be ascertained . Presence of celluase even after through
cleansing of gut lumen by flushing indicates that the cellulase in question is perhaps not bacterial in origin
(Debnath et.al. , 1989) . Because , so far the amount of concentration is concerned , cellulase stands next
to amylase (Table 30) . That Whether this should be regarded as a preadaptation for its omnivorous habit
or as retention of its ancestral enzymic complement is not clear . Fage (1949) reported the hepatopancreas
to be the site of enzyme production and digestion , the digestion being not entiraly extracellular , but also
occurring intracellularly in hepatopancreas (Schlottke , 1934a,b) . This is further coroborated by the
present observation that most of the hydrolysing enzymes are present in large amount in the hepatopancreas
A synopsis of horseshoe crab's food-items with special reference to L. polyphemus is given
below ( based on Shuster 1982 ) :

1. Trilobite stage : Experimental feeding with a variety of organic matter and organisms e.g. bits of
Mytilus , Nereis , frozen brine shrimps , marine nematodes , small whole polychaetes , and alga ,
Enteromorpha sp. (French , 1979) .

2. Immature crabs : Worms - Cerebratulus , Nereis and Cistenids (Shuster , 1950) ; Bivalves -
Mya , consumption of 99 items in 72 hours (Turner et. al. , 1948) ; Macoma , Gemma , Ensis , Spisula
and Mytilus (Shuster , 1950) .

3. Mature (adult crabs) : Comparatively soft-shelled molluscs , both gastropods and bivalves ; annelids
(mainly polychaetes) , arthropods , plant materials and miscellaneous category including bryozoa ,
nematodes , teleosts , turbellarians and foram shells . (Shuster , 1982 ; Botton , 1981 ; 1984a,b etc.) A
considerable amount of ophiuroids reported for the first time T. gigas as food item (Debnath and
Choudhury , 1987; Debnath et.al. , 1989) .

NEXT
CHAPTER
ON

CENTRAL
NERVOUS SYSTEM

&

NEURO-ENDOCRINE
SYSTEM
 134

PART - III
STUDIES ON ANATOMY AND CYTOMORPHOLOGY
OF THE CENTRAL NERVOUS SYSTEM
WITH SPECIAL REFERENCE TO THE
NEURO - SECRETORY ELEMENTS

Olfactory nerve
Optic nerve
Cheliceral nerve
PROTOCEREBRUM
(Supra-pharyngeal ganglion)

TRITOCEREBRUM
(Circum-oesophgeal Ring)

ORAL COMMUSSURE

Nerves to walking legs

Nerve to 6th walking legs

SUB-PHARYNGEAL
GANGLION
VIII VENTRAL NERVE
CORD
Stomodeal nerve
Nerve to genital operculum
IX

X
Nerves to book-gills
XI
XII
XIII
XIV

Caudal nerves

Central Nervous System of Horseshoe Crab,

Carcinoscorpius rotundicauda
 135

INTRODUCTION

Research literature on arthropod nervous and neuroendocrine system is immense (Bullock and
Horridge , 1965 ; Gabe , 1966 ; Highnam , 1965 ; Novak , 1975) . The organization of the arthropod
nervous system has a striking correlation with their increased cephalization . This cephalization in
arthropods is seen not only in greater abundance of cephalic sensory organs and their specialisation but is
also reflected in a more highly developed brain . The arthopod brain , in general , consists of three major
regions - an anterior protocerebrum , a median deutocerebrum and a posterior tritocerebrum . Of these
three , the deutocerebrum receives the antennal nerves ( 1st antennae of crustaceans) and contain their
associated organs. Antennae are lacking in the chelicerates like merostomates (horseshoe crabs) and
arachnids (spiders and scorpions). As a result the chelicerate brain is devoid of the deutocerebrum . The
brain in arthropods is joined to a ventral cord via circum-oesophageal connective . Arthropod nervous
system , like those of annelids contains many sensory cells and interneurones . The flexibility of arthopod
behaviour and their impressive power of association learning , may thus seem surprising . Part at least of
the explanation , however , is doubtless , to be searched in the extensive branching of their nerve
processes , and the variety of connections thereby made possible , so that the nervous system is very much
more flexible in action .
The nervous system in all invertebrates contains in addition to "ordinary" or " typical"
neurones (as concerned with the propagation of nerve impluses and the secretion of chemical transmitter
substances) a second type of neurone containing secretory product that is readily stainable and that can
be seen with the light microscope . It is referred to as neurosecretion . This material , like the
neurohumours , arises in the cell body , passes down the axon , and is released from the nerve endings .
After being discharged from the cells it passes into the blood stream and circulates round the body , to
produce specific physiological effects at targets that may be remote from the region of its release . The
cells producing this substance are customarily referred to as neurosecretory cells ; their function is ,
thus , the secretion of neurohormones , which form part of the endocrine system of the body and thereby
termed as the "neuro-endocrine ststem" (Barrington , 1979) .
Among the arthropods most of the work on neurosecretion are confined to the mandibulates
like Insecta and Crustacea (Bliss and Welsh 1952 ; Matsumoto , 1954 ; Kobayashi , 1957 ; Highnam ,
1965 ; Madrell , 1967 ; Novak , 1975 ; Ramadon and Matta , 1976 ; Nanda et. al. , 1984 ; Ghosh and
Nanda , 1985) , but information is scanty and unsatisfactory regarding the neurosecretion in Chelicerate
arthropods (Scharrer , 1941 ; Gabe , 1955 ; Enami , 1957 ; Habibulla , 1961 , 1970 ; Herman and Preus ,
1973) . The earliest report of neurosecretion in Merostomata is that of B . Scharrer (1941) who reported
the occurrence , localozation and quantitative distribution of "putative neurosecretory cells" throughout
the central nervous ststem of Limulus polyphemus and compared the findings with Tachypleus
gigas(collected from Penang , Malayasia by the author as "Limulus moluccanus"in her paper). Later ,
Enami , 1957 ; and Herman and Preus (1973) reported two types of neurosecretory cells in rudimentary
eyes of both L. polypheus and T. tridentatus and the abdominal ganglia of L. polyphemus
respectively , on the basis of cell dimension and cytoarchitecture .
The present investigation deals with anatomy and cytomorphology of the central nervous
system (CNS) and neurosecretory cells (NSCs) of Carcinoscorpius rotundicauda which has hitherto
not been recorded so far . Such studies are necessary to correlate the event of histophysiological changes
brought about by environmental stimuli. Since all the functional activities of an arthropod , like breeding
and feeding behaviours , development , growth , molting , circadian rhythmicities , dispersal and migra-
tion etc. , are directly or indirectly controlled by its neuroendocrine system , the present investigation ,
obviously , is expected to highlight a new area of study on Indian horseshoe crabs .
 136

OBSERVATIONS

(A) Anatomy of the CNS

The nervous system displays (Fig. 33A) a large degree of fusion plus the very unique feature of
being almost entirely enclosed within arterial vessels (hence the term " vascular-ring") . The brain forms
a collar around the oesophagus . The anterior part (suprapharyngeal ganglion) of the collar forms the
protocerebrum , while the lateral portions (subpharyngeal portions) represent a fusion of the tritocerebrum
plus the ganglia for all the remaining first seven segments . The collar circumscribes and unites behind the
oesophagus , giving rise to both a ventral nerve cord with five ganglia and lateral nerves extending through
the abdomen .
The brain receives nerves from two pairs of lateral eyes as well as from the frontal organ (i.e.
the olfactory nerve). The tritocerebrum fuses with the posterior part of the brain and receives the cheliceral
nerves . Ganglia II to VIII have moved forward and fused to a large sub-pharyngeal mass , serving not only
the prosoma but also abdominal segments VII and VIII . Abdominal ganglia IX to XIV stay in their respec-
tive segments and form a nerve chain , the anterior ones close to the appendages , the last three in one
mass , possibly a vestige of the 15th segment .
So , in general , the nerve ring consists of 3 or 4 post oral commissures on the dorsal part and 1
pre-oral commissure on the ventral side at the posterior margin , an anterior cerebral swelling , the nerves
to the appendages , the stomodeal nerves , the olfactory nerves , and the nerves to the median and lateral
eyes .

(B) Cytomorphology of CNS

The CNS is covered by an outer thick connective tissue sheath and the neurosecretory cells
(CNCs) are ordinarily distributed throughout the CNS . The NSCs are mainly concentrated in groups
extending from ventral to dorsal surface of each ganglia and the arranged symmetrically all around the
central fibrous neuropile with respect to the midsagittal plane . It is to be noted that the NSCs stain
faintly with CHP and AF , moderately with Masson's trichrome and intensely with Azan method . On the
basis of their size , cytoarchitecture and pattern of distribution in the CNS , the NSCs may be classified
into two major types : namely, A and B cells .

A-cells : These cells are considerably large (diameter ranging from 90 to 100 µm ) , spherical or
pear-shaped in appearence ( Fig. 33 B&D) and possess moderately stained cytoplasm . Axonal process
may or may not be present . Such type of cells are often distributed beneath the perineurium and usually
form groups . A-cells is characterised by the presence of a centrally placed conspicuous round nucleus
measuring about 20-22 µm in diameter besides possessiing well defined nucleous (Fig. 33C) . The
cytoplasm exhibits clumpy appearence and indications for having cyclical secretory activity ranging from
a homogeneous condition to a state of fine and occasionally coarse granulation . Strikingly , majority of
the cells possess a ring of cytoplasmic vacuoles in between the cytoplasmic clump and the cell membrane
(Fig. 33E) . Axonal bundle ( Figs. 33 E and F) . Seldom transport of neurosecretory material through the
axons could be visualised .
 137

B-cells : Cells of such types are smaller with wide range of fluctuation in their diameter (30 to 50
µm ) . They are irregular in outline and generally exhibit deep stained cytoplasm having coarse consist-
ency (Figs. 33 B and D) and possess centrally placed prominent nucleus (12 -15 µm ) . These cells are
more numerous than A-types and ordinarily distributed , very close to the neuropile forming compact
groups of cells . Axonal processes are generally long , filamentous , deeply stained and frequently form-
ing large axon bundles (Fig. E & G) .

Discripancies in the pattern of distribution of NSCs in different parts of CNS

A and B cells are deterctable in all the ganglionic parts of the CNS . But their distribution
varies with respect to different ganglia . The posterior third of the circumoesopageal ring (containing
fused 2-3 thoracic ganglia) shows dramatic increase in number of NSCs when being compared with the
anterior two third of the ring . The abdominal ganglia also retains increased population of NSCs .

Brain : B-cells (mean diameter 35 µm ) are much more in number than the A-cells (mean
diameter 90 µm ) and form compact cell groups (Fig. 33 B) . Axon bundles emerging out from these B-
cell groups make a configuration of decussation within the neuropile (Fig. 34 B) . Well defined capillary
networks that supply the brain are found to encricle the neurosecretory cell groups and even the solitary
cell also ( Fig. 33C , 34A) . Besides these NSCs , there are large clusters of conspicuous small cells
with delicate axonal processes that are found all around the neuropile in the antero-median portion of
the brain . These cells , however , do not show any sign of secretory activity and probably these cells
are the ordinary neurones (Fig. 34C) .

Thoracic ganglia : A drastic enhancement in the number of both types of NSCs is noticeable .
Three important features are on record when compared with the brain NSCs :

1. Both categories of cells maintain larger dimension (A-cell : 100 µm , B-cell : 40µm ) .

2. Number of A-cells is relatively enhanced (Fig. 33D and F) .

3. Distinct of specific groupings of A- and B-cell types are demonstrable despite existence of
mixed groups comprising both A - and B - types .

Axnal bundles are , however , common in both categories of cells ( Fig. 33 E & G) . Besides
these , involvement of blood capillaries in respect of NSCs remain similar to that of brain .

Abdominal ganglia : The abdominal ganglionic NSCs in respect of diameter , number , groupings ,
supply of blood capillaries etc. have close resemblance with the NSCs displayed in the thoracic ganglia
(Fig. 33 F and G) . However , a few A-cells show giant size (100-120 µm ) with coarse cytoplasm , large
peripheral vacuoles and long tubular axonal processes (Fig. 34A) . Axon bundles are more abundant in the
abdominal ganglia in comparison with other regions of CNS .

A comparative account of size range and staining affinities of the NSC types in the CNS of
Carcinoscorpius rotundicauda are given in Table 31 .
 138

Table 31

Comparative size ranges and staining affinities of the two types of BSCs
in the central nervous system (CNS) of Carcinoscorpius rotundicauda .

ø
Cell Mean cell Mean nuclear Staining affinity of NSM
type diameter(µ m) diameter (µ m)
CHP AF Masson's Trichrome Azan

Brain

A 90 20 + + ++ +++

B 35 12 + +/- +++ ++++/+++

Thoracic
ganglion

A 100 22 + + ++ +++

B 40 15 + + ++ +++

Abdominal
ganglion

A 98 22 + + ++ +++

B 38 12 +/- + +++ ++++/+++

ø NOTE : + = weak/ faintly positive , ++ = moderately positive ,

++++/+++ = strong (intensely positive) , +/- = doubtful reaction
 139

Central nervous system

of C. rotundicauda

B C

D E

A F G

Fig . 33 (A-G)

A, Central nervous system (CNS) of C. rotundicauda showing the circumoesophageal ring

(brain) and the ventral nerve cord with emerging nerves from them ; B. Section of brain of C.
rotundicauda showing distribution of two types of NSCs ( A & B) , Azan x 125 ; C , Section of
brain showing individual A cells encircled by blood capillaries , Azan x 532 ; D , Section of
thoracic ganglion , showing an increase in the number of A cells . Azan , x 125 ; E , Section
showing groups of A cells and B cells forming a large axon bundle , Masson's trichome , x 125 ;
G , Section of abdominal ganglion showing a large group of A cells , Azan , x 125 .
 Blood Capillry Neuro-secretory
granules
B-cells of the Brain Peripheral Vacuole

A B

B Central
C
Decussation of Nucleus
axon bundles
Nucleolus

Atypical Neurones

Axon bundle

Fig . 34 (A-C)

A, Shows " decussation" of axonal bundles emerging from the B cells

in the brain region ; B , Shows an enlarged cell-body of A-type displaying
peripheral vacuoles containing NSM , central rounded nucleus with a
distinct nucleolus . The cell-body is encircled by a capilary system
(dark) , Azan , x 1100 ; C , Aggregation of numerous minute ordinary
( "atypical") neurones associated with cellular bodies of NSCs .
 141
DISCUSSION

Histological probe of the central nervous system of Carcinoscorpius rotundicauda shows the
existence of major two types of cells which maintain characteristic pattern of distribution within the
circumoesophageal ring , as well as , the ventral nerve cord . In her pioneering work B. Scharrer (1941)
reported a single type of neurosecretory cell throughout the CNS of Limulus polyphemus . She charac-
terised these cells on the basis of the possession of large colloidal mass , marginal vacuolations and
eccentric nuclei . In contrast with this observation , Herman and Preus (1973) observed 2-types-Type I
and Type II of NSCs in the same species . These observations , however , based on both light and electron
microscopic studies and chiefly restricted to the morphological characteristics of the granules despite
other cytomorphic features . In the present investigation on C. rotundicauda the author did not observe
coloid laden NSCs throughout the CNS as reported by B. Scharrer (1941) . On the other hand , two types
of NSCs ( A- and B-types) are encountered and classified on the basis of their distribution , shape , size
and other morphological features and is in agreement with the observation of Enami (1957) , Herman and
Preus (1973) , as and when , morphological features of cell types as such is taken into account . It is
interesting to record that like the ventral ganglionic 'C' cells of mandibulates neurosecretory elements of
horseshoe crabs are readily identifiable with Azan , as well as Masson's trichome stains but react poorly
with either CHP or AF . This may have some bearing with the histochemical nature of the neurosecretory
product in the Xiphosura which possibly retains its strong acidopholic nature that could not be scuttled
after oxidation of tissue section with potassium permenganete (Gabe , 1966) .
In Carcinoscorpius differential pattern in the distribution of NSCs has been recorded in the
CNS . The most "active" neurosecretory region in CNS is posterior part of the circumoesophageal ring .
Here the neuroglandular elements are found in clusters that occupy a considerable portion of the
ganglia . B. Scharrer (1941) who reported maximum concentration of chromatophototropic principle in
posterior third of the Limulus circumoesophageal ring . Increased population and compact arrangement
of NSCs and occurrence of more axonal bundles in the abdomical ganglion are all indicative of its func-
tional importance when correlation between physiological and anatomical findings are taken into account
(B. Scharrer , 1941) .
Although occurrence of well defined axon bundles is a regular feature in the neurosecretory
system of Carcinoscorpius rotundicauda , transport of discrete stainable material (NSM) through
them , is seldom .It is striking that despite the presence of open type of circulation in Xiphosura , capillar-
ies are found to surround neurosecretory cells throughout the CNS of Carcinoscorpius . Similar ar-
rangement of capillaries around the NSCs had also been reported in the central nervous system of crabs
like Paratelphusa hydrodromus (Parameswaran , 1956) and Eriocheir japonicus (Matsumoto ,
1954) . These observations indicate that the neurosecretory materials in Carcinoscorpius sp. are possi-
bly discharged directly from the perikarya into the blood and thus reach the general circulation of the
body readily without undergoing axoplasmic flow or terminating to the conventional neurohaemal
organs . Enami's (Enami , 1957) and our's (Choudhury et. al. , 1991) similar findings suggest that the
ventral arteries surrounding the central and peripheral nervous systems play a role in transportation of
the NSM , instead of the axons as in many other animals . Such an arrangement of the circulatory system
in the CNS also suggests special functions in the event of attributing the species under the category of
"Living fossil" (Choudhury et. al. , 1991) . Nevertheless , neurosecretory cells must be expected to have
a high metabolic rate and , therefore , require a rich blood supply . This is also the case in vertebrates
where nuclei composed of secreting nerve cells are among the most richly vascularised structures of the
central nervous system (Scharrer and Scharrer 1954) .
In fine , it is to be noted that the functional activities of the neuroendocrine system of horse-
shoe crabs are , probably , attributed with the moulting phenomena (jegla , 1972 ; Jegla and Costlow ,
1979a,b ; Krishnakumaran and Sneiderman , 1970) , visual system and mating behaviour.
 142

(Barlow , 1982 , 1983 ; Barlow et. al. , 1977 , 1982 , 1984 , 1985 , 1986 , 1987 ; Lall and Chapman ,
1973 ; Rudloe , 1979b , 1980) and other behavioral patters (Botton and Loveland , 1987) . It is also quite
possible to control the feeding behaviour as was observed in the red cotton bug , Dysdercus cingulatus
that the median neurosecretory cells of the brain are responsible for the stimulation of food consumption
and respective enzyme activity through "Secretogogue mechanism" (Muraleedharan and Prabhu , 1979) .
Similarly Thomsen and Moller (1959) reported the role of median neurosecretory cells in digestion of
Calliphora erythrocephala . The presence of two putative neurohormones (dores and Herman , 1980) in
the CNS of L. polyphemus might have some correlation with the present observations.
Nucleolus

Blood Capillry
Cytoplasm

Nucleolus
Cytoplsmic vacuoles
Sec
ret

Central Nucleus
ory

Spherical
Gra

Nucleus
nul
es

Axoplasm
on
Ax

A - type NSC of Thoracic Ganglion A - type NSC of Brain

Since all the functional activities

of an arthropod , ...................... are
directly or indirectly controlled by
its neuroendocrine system , the
present investigation , obviously ,
is expected to highlight a new area
of study on Indian horseshoe crabs
 143

SUMMARY of THE THESIS

The present study on geographic distribution confirms that the western limit of distribution
of the two species of Indian horseshoe crabs ,Tachypleus gigas(Muller)and Cacinoscorpius(=Tachypleus
n.comb.) rotundicauda (Latreille) is up to the coastal zone of Orissa (lake Chilka) and West Bengal
(Sundarbans) through Bangladesh . Discrete local population of T. gigas occurs in the sandy sea shores .
C. rotundicauda dominates the muddy shores of Sundarbans and Dhamra estuaries , while T. gigas also
occurs at the later area showing typical sympatrism with C. rotundicauda . The present work also
confirms that T. tridentatus does not exist in the Indian coast .
Comparative morphology as well as morphometry reveal that the male is nearly 78% and 85%
of the female of T. gigas and C. rotundicauda , respectively . But in respect to body weight the male
T. gigas is almost 37% of the female , while it is 52% in C. rotundicauda .Considering all the
morphometric parameters it is found that the female specimens of C. rotundicauda are smaller than the
females of T. gigas , but the situation is just reversed in case of male specimens . It is also evident that a
perfect correlation occurs between the dorsal body length (DBL) and the dorsal prosoma breadth
(DPB) . So it seems to be justified to make a length - weight relationship taking count the DBL as the
only size parameter . In this study , the male increases in weight by an exponent of 3.08 and the female by
2.52 in T. gigas , while in C. rotundicauda males and females increase by an exponent of 3.09 and
2.35 , respectively . The general growth patterns are found to be more or less identical in both the
species . The marking study reveals that nearly 5,000 T. gigas come ashore at Chandipur (Orissa) during
the fullmoon tides of March , 1985 . The total absolute estimate is obtained after regression
method to be nearly 70,000 during the entire breeding season that ranges from March to July
(Chandipur) .This figure is rather small i.e. almost 2,000 in the sea shore area of Digha (West
Bengal) .
During fullmoon high tides of spring and summer months horseshoe crabs are found to emerge
from the shallow sea to spawn . The usual 1:1 sex-ratio is obtained for the occurrence of normal amplexus
i.e. the larger female carrying a smaller male on her back . Very frequently male biased sex-ratio is found
due to stranded male crabs occurring at the exposed sandy beach or mud-flat . It is evident from the
present study that deviation in the sex-ratio is significant in all coastal stations of West Bengal , while
it is not statistically significant at the coast of Orissa . This deviation in sex-ratio (6.89 :1) at Digha
than of Chandipur (1.04:1) might be attributed to the severe predatory pressure exerted by the
ravens , Corvus splendens L. and human interference .
Preliminary studies on the feeding habit and digestive physiology of T. gigas has indicated its
omnivorous food habit . Gut content analysis also enumerates a varieties of food matters e.g. vascular
plant parts , bivalves , gastropods , polychaetes , ophiuroids , amphipods and several others to infer the
horseshoe crabs also as a " dietary generalist ."
Study on central nervous system of C. rotundicauda has revealed two types of neurosecretory
cells ( A & B ) differing as regards to their size , cytoarchitecture and distribution . The A cells are
large , with fine cytoplasmic granules (NSM) , and are distributed below the perineurium , while the B
cells are much smaller , contain coarse granules and are distributed in the vicinity of the neuropile . The
posterior third of the circum-oesophageal ring and the abdominal ganglia contain a far larger number of
NSCs as part of the CNS . In fine , it is to be noted that the functional activities of the neuroendocrine
system of horseshoe crabs are , probably , attributed with the moulting phenomena , visual system and
mating behaviour.
 144
REFERENCES

Allee , W.C. 1923 . Studies in marine ecology II . An annotated of the distribution of common inv-
ertebrates of the Woods Hole littoral.Marine Biological Lab .Woods Hole, Massachusetts.

Andrewartha , H.G. 1970 . Introduction to the study of animal population . 2nd Edn. ELBS & Chapman
and Hall Ltd. , London , U . K .
Andrews , E.A. and Nelson , T.C.1942. A folliculinid carried by Limulus Anat. Rec. 84 : 495
Annandale , N.1909 . The habit of King crabs , Rec. Ind. Mus. 3 : 294-295 .
Annandale , N.1922 . The marine element in the fauna of Ganges . Bijdr. Dierk. 143-154 .
Asano , U.1942 . On the life history of horseshoe crabs . Bot. and Zool. 10 : 120-124 (in Japanese) .
Bang , F.B.1956 . A bacterial disease of Limulus polyphemus . Johns Hopkins Hosp. 98 : 325-351 .
Bang , F.B.1979 . Ontogeny and phylogeny of response to gramnegative endotoxins among the marine
invertebrates in Cohen , E et.al. , (eds) . Biomedical Applications of the Horseshoe Crab
(Limulidae) : Progress in Clinical and biological research . vol. 29. Alan , R. Liss . New
York , pp. 109-123
Banner , A.H. and Stephens , B.J. 1966 . A mote on the toxicity of the horseshoe crab in the Gulf of
Thailand . Nat. Hist. Bull. Siam . Soc . 21 : 197 -203
Baptist , J.P. 1953 . Record of hermaphroditic horseshoe crab , Limulus polyphemus L. Breviora,
14 : 1-4
Baptist , J.P; O.R . Smith and Ropes , J.W. 1957 . Migrations of the horseshoe crab , Limulus
polyphemus , in Plum Island Sound , Masschusetts , U.S. Fish. Wild. Svc. , Spec. Sci. Rep.
Fish . no. 220- : 1-5
Barber , S.B. and Hayes , W.F. 1963 . Properties of Limulus chemoreceptors . Proc. XVI. Int.
Cong. Zool. (Washinton , D.C.) pp. 76-78
Bailey , N.T.J. 1951 . On estimating the size of mobile population from recapture data . Biometrika ,
38 : 293-306
Bailey , N.T.J. 1952 . Improvements in the interpretation of recapture data . J. Anim. Ecol. 21: 120-127
Bailey , N.T.J. 1959 . Statistical methods in Biology . ELBS , London , UK.
Bargmann , W.1949 .Uber die neurosekretorische Verknupfung von Hypothalamus and Neuropophyses .
S. Zellforsch. 34 :610
Barlow , R.B. , Jr. 1982 . Seasonal changes in the circadian modulation of sensitivity of the Limus
lateral eye . Biol. Bull. 163 : 380
Barlow , R.B. , Jr. 1983 . Circadian rhythms in the Limulus visual system . J. Neurosci . 3 : 856-870.
Barlow , R.B. , Jr., Bolanowski , Jr. , and Brachman , M.L. 1977 . Efferent optic nerve fibres mediate
circadian rhythms in the Limulus eye. Science . 197 : 86-89
Barlow , R.B. , Jr. Ireland , L.C. and Kass , L. 1982 Vision has role in Limulus mating behavior
Nature . 296 : 65-66
Barlow , R.B. , Jr. Kaplan , E. , Renninger , G.H. and Saito , T. 1985 . Efferent control of circadian
rhythms in the Lumuls lateral eye . Neurosci . Res . Suppl . 2 : 565-578
Barlow , R.B. , Jr. and Palpai , M.K. , Howward , H. and Kass , L. 1986 . Migration of Limulus for
mating : Relation to lunar phase , tide height , and sunlight . Biol . Bull . 171 : 310-329
Barlow , R.B. , Jr., Powers , M.K. , and Kass , L. 1987 . Vision and mating behavior in Limulus.
chapter 17 . in Sensory Biology of Aquatic Animals . Popper , A. R. Fay , and J. Atema (ends)
Springer - Verlag , Berlin .
Barlow , R. B. Jr. Powers , M.K. , Kass , L. Fiordalice , R.W. , Camara , M.D. and Howard , H.A.1984.
Vision in Lumulus mating behavior during day and night . Biol . Bull . 167 : 522
 145

Barrington , E.J.W. 1979 . Invertebrate structure and function , 2nd Edn. ELBS , Great Britain , p. 765

Barthel , K.W.1974 Limulus : A living fossile . Horseshoe crab aids interpretation of an Upper
Jurassic environment (Solnhfen). Naturwissenschaften , 61 : 428-433
Beamont , W.B.S.1884 . On the testis of Lumulus . Trans . Linn. Soc . London . (Zool) , 2 :362-366
Benham , W.B.S.1885 . On the muscular and endoskeletal systems of Lumulus and Scorpio .Trans .
Zool. Soc. London , 11 : 311-384
Bennett , D.C. , Pezella P.F. and Herman , W.S. 1972 . A sexual mosaic with internal embryogenesis
in the diocious horseshoe crab , Limulus polyphemus . Bijd. Dierkd . 42 (2) : 192
Bernfeld , P. 1955 . Methods in Engymology , Colwick , S.P. and Kaplan , N.O. (ends) . Acad . Press ,
New York 1 : 149
Bishayee , S. and Thambi Dorai , D. 1980 . Isolation and characterization of a sialic acid binding
lectin (carcinoscorpin) from Indian horseshoe crab , Carcinoscorpius rotundicauda .
Biochem . Biophys . Acta . 623 (1) : 89-97
Bishop , J.A. and Sheppard , P.M. 1973 . An evaluation of two capture recapture models using the
technique of computer simulation in Bartlett , M.S. and R.W. Uirons (ends).The Math -
ematical Theory of the Dynamics of Biological populations . pp. 235-252 , Academic
Press , London
Bliss , D.E. and Welsh , J.H. 1952 . The neurosecretory system of brachyuran Crustacea . Biol . Bull.
103 : 157-169
Bonaventura , J . , Bonaventura , C. and Tesh S. (ends), 1982 . Physiology and Biology of Horseshoe
Crabs : studies on Normal and Environmentally Stressed Animals . Progress in Clinical
and Biological Research . (vol. 81) Alan R. Liss , Inc. New
Botton , M.K. 1981 . Food habits of breeding horseshoe crabs in Delaware Bay . Bull. N.J. Acad . Sci.
26 : 68
Botton M.L.1982 . Patterns of abundance of the horseshoe crab , Limulus polyphemus , in Delaware
Bay and coastal New Jersey . Bull . N.J. Acad . Sci . 27 : 39
Botton M.L.1984a . The importance of predation by horseshoe crabs , Limulus polyphemus , to an
intertidal sand flat community . J. Mar . Res. 42 (1) : 139-162
Botton M.L.1984b. Diet and food preference of the adult horseshoe crab , Limulus polyph in
Delaware Bay , New Jersey , USA . Mar. Biol . 81 : 199-207
Botton M.L.1984c. Effects of laughing gull and shorebird predation on the intertidal fauna at Cape
May , New Jersey (USA) . Coastal Shelf Sci . 18 (2) : 209-220
Botton M.L.1984d. Spatial distribution of 3 species of bivalves on an intertidal flat : The interaction of
life history strategy with predation and disturbance . Veliger 26 (4) : 282-287
Botton , M.L.1984e. What determines the vulnerability of bivalve prey to horseshoe crab predation?
Anonymous J. Shellfish Res . 83-84 : 1982 . Annual Meeting of the National shellfisheries
Association , Baltimore , Maryland , USA . (June 14-17 , 1982)
Botton M.L. and Haskin , H.K. 1984 . Distribution and feeding of the horseshoe crab , Limulus
polyphemus , on the continental shelf of New Jersey . Fish . Bull . 82(2) : 383-389.
Botton M.L. and Loveland R.E. 1987. Orienatation of horseshoe crab Limulus polyphemus , on the
sandy beach . Biol . Bull 173 (2) : 289-298
Botton M.L. and Loveland R.E. 1989 . Reproductive risk : high mortality associated with spawning
by horseshoe crabs ( Limulus polyphemus) in Delaware Bay (USA) .Mar . Biol. 102 (2) :
143-152
 146

Botton M.L. , Loverland , R.E. and Jacobson , T.R. 1988 . Beach erosion and geochemical factors :
influence on spawning success of horseshoe crabs ( Limulus polyphemus) in Delaware Bay
(USA) . Mar. Biol. 99 (3) : 325-332
Botton , M.L. and Ropes , J.W. 1987. Populations of horseshoe crabs , Limulus polyphemus , on the
north-western Atlantic continental Shelf . U.S. Natl. Mar. Fish . Surv. Fish . Bull . 85 94) :
805-812
Botton , M.L. and Ropes , J.W. 1988 . An indirect method for estimating longecity of the horseshoe
crab (Limulus polyphemus) based on epifaunal slipper shells ( Crepidulafornicata) J . Shell
fish Resh. 7 (3) : 407-412
Bolau , H. 1879. Moult of Limulus polyphemus (Lat.) described . Verh Ver Naturw Unterh Hamburg,
1879 :22-23
Brockmann , H.J. 1990 . Mating behaviour of horseshoe crabs , Limulus polyphemus Behavior , 114
: 206-219
Brown , G.G. 1979 . Scanning electron microscopical and other observations of sperm fertilization
reactions in Limulus polyphemus L. (Merostomate : Xiphosura ) .J.Cell . Sci. 22 : 547-562
Brown , G.G. and Knouse , J.R. 1973 . Effects of sperm concentration , sperm aging and other
variables on fertilization in the horseshoe crab , Limulus polyphemus , Biol . Bull. 144 (3) :
462-470
Brown , G.G. and Humphries , W.J. 1971 . Sperm-egg interactions of Limulus polyphemus with
Scanning electron microscopy J. Cell . Biol . 51 : 904-907
Brown , G.G. and Clapper , D.L. 1980 . Cortical reactions in inseminated eggs of the horseshoe
crab, Limulus polyphemus L . Dev. Biol. 76 : 410-417
Brown , G.G. and Clapper , D.L. 1981 . Procedures for maintaining adults , collecting , gametes and
culturing embryos and juveniles of the horseshoe crab , Limulus polyphemus L. in Labora
tory Animal Management : Marine Invertebrates . National Academic Pres, Washington , D.C.
Brown , J.A. and Cunningham , 0 . 1941 . Upon the presence and distribution of a chromatophorotropic
principle in the central nervous system of Limulus . Biol. Bull. 81 : 80
Bullock , T.H. and Horridge , G.H. 1965 . Structure and function in the nervous system of Inverte -
brates. 2 vols . Freeman , San Francisco .
Cameron , M.L. and Steele J.E. 1959 . Simplified aldehyde fuchsin staining of neurosecretory cells .
Stain . Technol . 34 :265-266
Cavanaugh , C.M. 1975 . Observations on mating behaviour in Limulus polyphemus . Biol. Bull. 149
: 422
Chamberlain , S.C. and Barlow , R.B. Jr. 1984 . Transient membrane shedding in Lumulus
photoreceptors : control mechanism under natural lighting . J. Neurosci . 4 : 2792-2810
Chanchal, A.K. , Pandey , B.N. and Sing , S.B. 1978 . Studies on some aspects of the biology of
Anabas testtiudineus (Teleosti : Anabantidae) . Matsya 4 : 15-19
Chatterjee , A. , Vijay Kumar , R. and Parulekar , A.H. 1988 . Growth and morphometric characteristic
in the horseshoe crab , Carcinoscorpius rotundicauda (Latreille) from Canning (West
Bengal) , India . Pak . J. Sci. Ind. Res. 31 (5) : 352-353
Cheng , T.C. 1982 . The re-emerging area of marine biomedicine (Limulus polyphemus etc.) J.
Invert . Pathol . 40 (2) : 155
 147

Choudhury , P.S. Nanda . D.K. and Debnath , R. 1991 . Cytomorphic studies on the neurosecretory
cells in the central nervous system of the Indian horseshoe crab Carcinoscorpius
rotundicauda (Latreille). Func. Dev. Morp. 1 (1) : 13-16
Chowdhury , S.H. and Hafizuddin , A.K.M. 1980 . Horseshoe crabs (Chelicerata : Merostomata)
occurring along the south eastern coast of Bangladesh . Bangladesh J. Zool. 8 (1) : 5-14
Clare , A.S. , Lumb , G. , Clare , P.A. and Costlow , J.D. Jr. 1990 . A morphological study of wound
response and telson regeneration in post - larval Limulus polyphemus (L). Invertebr .
Reprod . Dev. 17(1) : 77-88
Cohen , E. , Bang, F.B. , Levin , J. , Marchalonis , J.J. , Pistole , T.C. , Prendergast , R.A. ,
Shuster , C.N. Jr. and Watson , S.W. (ends) 1979 . Biomedical Applications of the Horseshoe
crab (Limulidae). Progress in Clinical and Biological Research , (vol. 29) Alan R. Liss ,
Inc. New York. , p. 688
Cohen , J.A. and Brockmann , H. J. 1983 . Breeding activity and mate selection in the horseshoe
crab , Limulus polyphemus . Bull. Mar. Sci. 33 (2) : 274-281
Dana , J.D. 1852 . Crustacea . U.S. Exploring Expedition , 13 (1) pp. 1393 , Philadelphia .
Debnath , R. 1985 . Studies on the distribution , abundance , breeding behaviour , and feeding
habit of Indian horseshoe crabs (Merostomata : Xiphosura) in the coastal water of
Orissa and West Bengal , India . M. Phil . Diss. (Unpubl.) University of Calcutta , Calcutta .
Debnath , R. 1987 . "Vanishing crabs " in Letters to the Editor . The Statesman (16th July) p. 8
Calcutta , India .
Debnath , R. 1991 . Studies on species morphology and sexual dimorphism of Indian horseshoe
crabs . Geobios . New . Reports . 10 : 128-131
Debnath , R. and Choudhury , A . 1985 . Breeding behaviour of Indian horseshoe crabs ,
Carcinoscorpius rotundicauda and Tachypleus gigas (Merostomata : Xiphosura) in
the Sundarbans estuarine complex , India . Natl. Symp. Biol . Util . Cons . Mangt.
(18-20th Nov . 1985) Contr. no. 89 : 45 (abstr.) Shivaji Univ . , Kolhapur , India .
Debnath , R. and Choudhury , A . 1987 . Studies on food preference and feeding habit of horseshoe
crabs (Merostomata : Xiphosura) in the coastal waters of West Bengal , India .Internatl.
Symp. Trop. Ecol . (IX ISTE , 11-16th Dec. 1987) , Contr . no. 54 : 38-39 (abstr.) ,
Banaras Hindu Unit . , Varanasi , India .
Debnath , R. and Choudhury , A . 1988a . Predation of Indian horseshoe crab , Tachypleus gigas
by Corvus splendens . Trop. Ecol . 29 (2) : 86-89
Debnath , R. and Choudhury , A . 1988b. Length-weight relationship of the living fossil ,
Carcinoscorpius rotundicauda (Latraille) (Merostomata : Xiphosura) of Sundarbans
estuary , India . Env. Ecol. 6 (4) : 923-927
Debnath , R . and Choudhury , A . 1988c . Population estimation of horseshoe crab Tachypleus
gigas by capture-recapture method at Chandipur sea shore (Orissa) , India . J. Mar . Biol .
Ass. India . 30 (1&2) : 8-12
Debnath , R. and Choudhury , A . 1989 . Geographic distribution of Indian horseshoe crabs
(Merostomata : Xiphosura) in the coastal water of Bay of Bengal , India . J. Bengal Nat.
Host. Soc . (New Series) , 8 : 30-35
Debnath , R. and Choudhury , A . 1991 . Comparative assessment of two breeding population of
crabs ( Tachypleus gigas (Muller) (Merostomata : Xiphosura) . Env. Ecol. 9 (1) :
163-166
 148

Debnath , R. and Choudhury , A . 1992a . Length-weight relationship of the living fossile , Tachypleus
gigas (Muller) (Merostomata : Xiphosura) of Digha coast (West Bengal) India. Acta . Zool.
Bulgarica , 44 : 70-74.
Debnath , R. and Choudhury , A . 1992b . Rare occurrence of tandem amplexus by male Indian
horseshoe crabs , Carcinoscorpius rotundicuada , with a comment on mating in Limulidae
(Merostomata : Xiphosura) . Adv. Biosci. 15 : 103 - 107 .
Debnath , R . , Nag , S.K. , Choudhury , A , Dasgupta , R. and Sur , R.K. 1989 . Feeding habit and
digestive phusiology of the Indian horseshoe crab , Tachypleus gigas (Muller) . Ind. J. Physiol.
Allied . Sci . 43 (2) : 44-49
Dorai , D. T. , Bachhwat , B. K. , Bishayee , S. , Kannan , K. and Rao , D. R. 1981 . Further
characterisation of the sialie acid binding lectin from the horseshoe crab Carcinoscorpius
rotundicuada . Arch. Biochem . Biophysl . 209 : 325-333
Dores , R. M. and Herman , W.S. 1980 . The localization of 2 putative neurohormones in the central
nervous system of Limulus polyphemus . Comp. Biochem . Physiol . Comp . Physiol . 67
(3) : 459-464
Dunton , S. 1953 . The horseshoe crabs are gathering along the Atlantic coast . Animal Kingdom, 56 :
79-83
Eldredge , N. 1970 . Observations on burrowing behaviour in Limulus polyphemus (Chelicerata ,
Merostomata) , with implications on the functional anatomy of trilobites Am. Mus . Novit .
2436 : 1-17
Eldredge , N. 1974 . Revision of the sub-order synziphosurina (Chelicerata , Merostomata) with
remarks to Merostome phylogeny . Am . Mus . Novit . 2543 : 1-43
Enami , J. 1957 . Xiphosura in an Introduction to Neurosecretion . pp. 54-57 . Kyodo-isho ,
Tokyo (in Japanese) .
Fage , L. 1949 . Class de Merostomaces . Traite de Zoologie (Grasse , P. ed.) Masson at Cie , Paris , 6 :
219-262
Finn , J. J. , Shuster , C. N. Jr. and Swan , B. L. 1990 Limulus spawning activity on Delaware Bay
Shores . Finn . Tech . Industries . Inc . New Jersey
Fisher , D. C. 1975 . Swimming and burrowing in Limulus and Mesolimulus . In Martinson (ed.)
Evolution and Phylogency of the Trilobite , Trilobitoidae and Merostomata . Fossils
and Strata No. 4 Oslo . Universitetsforlaget , pp. 281-290
Fisher , D.C. 1977. Functional significance of spines in pennsylvanian horseshoe crab , Euproops
danae . Paleobiol . 3 : 175-195
Fisher , D. C. 1982 . Phylogenetic and macroevolutionary patterns within the Xiphosurida . J.
Paleontol . 56 (2) : 10
Fisher , D.C. 1984 . The Xiphosurida : archetypes of bradytely ? In : Eldredge , N. and S.M.
Stanly (eds) Living fossile . Springer Verlag , New York , p. 196-212
Fletcher , B.S.1973 . The ecology of natural population of the Queensland fruit fly , Dacustrvoni
IV . The immigration and emigration of adults . Austr. J. Zool . 21 : 541-565
Fowler , H.1908 . The king crab fisheries in Delaware Bay , and further note on New Jersey fishes ,
amphibians and reptiles. Annu . Rep. N.J. State Mus . (1907) 3 : 349-432
Freenkel , G. 1960 . Lethal high temperatures for three marine invertebrates : Lumulus
polyphemus , Littorina littorea and Pagurus longicarpus . Oikos , 11 : 171-182
French , K.A. 1979 . laboratory culture of embryonic and juvenile Limulus , in Cohen , E. et. al. ,
(eds) . Biomedical Application of the Horseshoe Crab (Limulidae) . Alan , R. Liss , New York .
pp. 61-71
 149

French , K.A. 1977 . An effect of light on the hatching pattern of Limulus polyphemus . Biol . Bull .
153 : 425
French , K.A. and Doliner , S. 1978 . Photopositive behaviour of young Limulus . Biol . Bull . 155
437-438
Gabe , M.1955 . Donnes histologiques surla neurosecre chez les Arachnides . Arach . Anat . Micr .
44 : 351-383
Gabe , M .1966 . Neurosecretion , Pergamon press , Oxford .
Gaskell , T.J. and George , B. J. 1972 . A Bayesian modification of the Lincoln Index . J . Appl. 9 :
377-384
Ghosh , P.K. and Nanda , D.K. 1985 . Cytomorphic studies of the thoracic ganolia of the prawn
Penaeus monodon(Fabricius) (Order : Decapoda ; Suborder : Macrura ; Family : Penaeidae) .
Folia Morph . 33 (3) : 299-303
Gomez Aguirre , S . 1979 . Notes for population studies of Likulus polyphemus L. (Xiphosura :
Xiphosuridae) at Islanel Carman , Campeche , Mexico . An . Inst . Biol . Univ . Nal . Auton .
Mexico . 50 , Ser . Zoolagica (1) : 769-772
Gorham , F. P. 1904 . Report of the special commission for the investigation of the lobster and soft
shell clam . III . The causes of death in artificially reased lobster fry . V.S. Comm . Fish.
Fisheries Rep . Comm . 1903 : 175-193
Goto , S. and Hattori , O . 1929 . Notes on the habits and growth stages of the Japanese king crab . Xe.
Congr . Intern . Zool . (Budapest) . 2 : 1147-1155
Gravier , C.1929a . Sur les caracteres sexual secondaires des Limulus . Compt. Rend. Acad . Sci.
189 : 1-14
Gravier , C .1929b. Revision de la collection des Limulus du Museem National d' Histoire Naturell .
Bull . Mus . Natl . Hist . Nat . Ser . 2(1) : 313-331
Groff , J. and Leibovitz , L. 1982 . A gill disease of Limulus polyphemus associated with triclad
turbellarian worm infection . Biol . Bull . 163 : 392
Gurr , E .1959 . A practical manual of medical and biological staining techniques . Leonard Hill (Books)
Ltd. London
Habibulla , M .1961 . Secretory structures associated with the neurosecretory system of the immature
scorpion , Heterometrus swammerdami . Q . J . Micr . Sci . 102 (4) : 475-479
Habibulla , M .1970 . Neurosecretion in the scorpion ( Heterometrus swammerdami) . J. Morph .
131 : 1-16
Hammen , V . D . 1986 . Comparative studies in Chelicerata : IV , Aptellata , Arachnida , Scorpionida ,
Xiphosura . Zool . Verh . (London) , O(226) : 1-52
Herman , W.S. and Preus , D. M. 1973 . Ultrastructural evidence for the existence of two types of
neurosecretory cells in the abdominal ganglia of the chelicerate arthropod , Limulus
polyphemus . J . Morph . 140 : 53-61
Howard , H. A., Fiordalice , R. W., Camara , M. D. , Kass , L., Powers , M.K. and Barlow , R.B. Jr.
1984 . Mating behaviour of Limulus : relation to lunar phase , tide height and sunlight . Biol .
Bull . 167 :527
Highnam , K.C.1965 . Some aspects of neurosecretion in arthropods . Zool . Jb. Physiol. 71 :
558- 582
Humm , H . J. and Wharton , G. W. 1942 . ecology of sand beaches at Beaufort , N.C. Ecol. Monogr.
12 : 135-190
 150
Ichishima , E.1970 . Methods in Enzymology Colowick , S.P. and Kaplan , N.O. (ends) , Acad .
Press , New York , 19 : 397
Lreland , L.C. and Barlow , R.B. Jr. 1978 . Tracking normal and blind folded Limulus in the ocean
by means of acoustic telemetry . Biol . Bull . 155 : 445-446
Ito Y . , Murai , M . , Teruya , T. , Hamada , R . and Sugimoto , A. 1974 . An estimation of population
density of Dacus cucurbitae with mark recapture methods. Res. Pop. Ecol .15: 213-222
Ives , J. E.1891 . Crustacea from the northern coast of Yucatan , the harbour of Vera Cruz , the
west coast of Florida and the bermuda Islands . Proc . Acad . Nat . Sci . Phila .43 :176-207
Iwanoff , P.P.1907 . Eiablage and larven von Limulus moluceanus . Bull . Dep. Agric . Inde.
Neerland. 8 : 18-221
Iwanoff , P.P. 1933 . Die embryonale Entwiek lung von Limulus moluceanus . Zool . Jahrb. (Anat.)
56 : 163-348
Jackson , C.H.N.1933a. On the true density of tsetse flies . J. Anim. Ecol. 2 : 204-209
Jackson , C.H.N.1933b. On a method of marking tsetse flies . J. Anim. Ecol. 2 :289-290
Jackson , C.H.N.1937 . Some new methods in the study of Glossina morsitans . Proc. Zool. Soc .
London , 1936 : 811-896
Jackson , C.H.N. 1939. The analysis of animal population . J. Arim. Ecol. 8 : 238-248
Jegla , T.C. and Costlow , J.D. Jr. 1970 . Induction of molting in horseshoe crab larvae by polyhydrxy
steroids . Gen . Comp. Endocrinol. 14 : 295-302
Jegla , T.C.1972 . Development and molting physiology horseshoe crab larvae . Am. Zool. 12
: 724
Jegla , T.C. and Costlow , J.D. Jr. 1979a. The Limulus bioassay ecdysteroids . Biol. Bull . 156 :
103- 114
Jegla , T.C. and Costlow , J.D. Jr.1979b. Ecdysteroids in Limulus larbae.Experientia, 35 :554-555
Jegla , T.C. and Costlow , J.D. Jr. 1982 . Temperature and salinity effects on developmental and early
posthatch stages of Limulus in Bonaventura , J. et.al. ,(eds.) Physiology and biology
of horseshoe crabs . Alan R. Liss . New York , pp. 103-113
Jegla , T.C. and Costlow , J.D. Jr. and Alspaugh , J .1972. Effects of ecdysones and some synthetic
analogs on horseshoe crab larvae . Gen. Comp. Endocrinol. 19 :159-166
BJerrison , H.J.1973. Evolution of the Brain and Intellegence ,Academic Press , New York .
Johansen , K . and Peterson , J.A. 1977. Respiratory adaptations in Limulus polyphemus (L) . In
Verenberg , F.J. (ed) . Physiological Ecology of Estuarine Organisms . pp . 129-145 .
University of South Carolina Press , Columbia .
Jolly , G.M.1965 . Explicit estimates from capture - recapture data with both death and
immigration - stochastic model . Biometrika, 52 : 225-247
Kampa , E.M.1970 .Underwater daylight and moonlight measurement in the eastern north Atlantic.
J . Mar . Biol. Assoc. U.K. 50 :397-420
Kaplan , E . and Barlow , R.B. Jr. ,Chamberlin , S.C. and Stelzner , D.J. 1976 . Mechanoreceptors
on the dorsal surface of Limulus . Brain Res. 109 : 615-622
Kawahare , D. 1982 . Investigations on ecology of horseshoe crab larvae . Aquabiology . 4 (5) :
380-382 . (in Japanese) .
Kawaguchi , S.1969. Report on protection and breeding of Tachypleus tridentatus in Kasaoka
Bay . 31 pp. Chugoku-Shikoku Agricultural Administration Bureau . (in Japanese)
Kawano , K. Yoneya , T. , Miyata , T. , Yoshikaea , K. , Tokunaga , F. , Terada , Y . and Iwanaga , S.
1990 . Antimicrobial peptide , tachyplesin I , isolated from hemocytes of the horseshoe crab
( Tachypleus tridentatus ) : NMR determination of the B-sheet structure . J . Biol.Chem .
265 (26) : 15365-15367
 151
Kawakatsu , M. and Sekiguchi , K. 1989 . Redescription of an ectoparasitic marine triclad ,
Bdelloura candida (Girard , 1850) Turbellaria ; Tricladida ; Moricola) . Collected from the
American horseshoe crab , Limulus polyphemus . Bull. Biogeogr. Soc. Japan. ,44 : 183-198
Kingsley , J.S. 1892 . The embryology of Limulus . J. Morphol . 7 : 35-68
Kingsley , J.S. 1893. The Embryology of Limulus , pt . 2. J.Morphol.8 : 195-268
Kishinouye , E.1892 . On the development of Limuluslongispina , J. Cel. Sci. Imp. Univ. Jpn. 5
: 53-100
Kobayashi , M.1957 . Studies on the neurosecretion in the silk-sorm , Bombyyx mori L. , Bull.
Ser. Expt. Stn. (Tokyo). 15 : 263-273
Koons , B. F.1883 . Sexual characters of Limulus , Amer . Nat . 17 : 1297-1299
Kooiman , P.1964 . J . Cell . Comp. Physiol. 63 : 197
Krishnakumaran , A. and Schneiderman , N .A . 1970 . Control of molting in mandibulate and
chelicerate arthropods by ecdysones , Biol. Bull . 139 : 520-538
Lafon , M.1941 . Sur la composition du tegument des crustaces , C.R. Soc. Biol. 135 : 1003-1007
Lall , A . and Chapman , R. 1973 . Phototaxis in Limulus under natural condition : evidence for
reception of nearultraviolet light in the median dorsal ocellus , J. Exp. Biol. 58 : 213-224
Latreille , P. A .1802 . Histoire Naturelle Generale et particuliere des Cristaces et Insectes . vol .3 .
Familles naturelles et generes . 467 pp. Dufact , Paris .
Laughlin , R.B.Jr. 1981 . Sodium chloride and water exchange in selected developmental stages of
the horseshoe crab , Limulus polyphemus , J. Exp. Mar. Biol. Ecol . 52 (2/3) : 135-146
Laughlin , R.B. Jr. 1982. The effects of temperature and salinity on larval growth of the horseshoe
crab , Limulus polyphemus , J. Exp. Blol. Ecol . 64 (1) : 93-113
Laughlin , R.B.Jr. . and Neff , J.M. 1977 . Interactive effects of temperature , salinity shock and
chronic exposure to No. 2 fuel oil on survival , development rate and respiration of the ]
horseshoe crab , Limulus polyphemus in : Wolff , D.A. (ed) . Fate and effects of petroleum
hydro carbons in marine organisms and ecosystems . Pergamon , Oxford , pp. 182 -194
Lazarus , R.J. and Reddy , P.S.R. 1986 . On the length weight relationship of the littleknown lesser
spiny Macrognathus aculeatus (Bloch) (Pisces : Mastacembeledae) , Proc. Ind.
Acad . Sci. B 95 (4) : 423-427
Leach , W.E.1819 . Dictionnaire des Science Naturelle , 14 : 537-538
Le Cren , E.D. 1951. The length-weight relationship and seasonal cycle in gonad weight and
condition in the pearch ( Perca fluvialis) , J. Animal. Ecol. 20 : 210-209

Leibovitz , L.1986 . cyanobacterial diseases of the horseshoe crab (Limulus polyphemus) , Biol.
Bull. 171 : 482-483
Leibovitz , L. and LEwbart , G.A. 1987 . A green algal (Chlorophycophytal) infection of the dorsal
surface of the exoskeleton , and associated organ structures , in the horseshoe crab,
Limulus polyphemus , Biol. Bull. 173 :430
Levi , H.W.1968. The name of the horseshoe crab is Limulus , Turtox News 46 (6) : 183

Lewis , A.E.1966 . Biostatistics . pp. 227 Litton Educational Publishing Inc. Pennsylvania , U .S . A .

Lincoln , F.C.1930 . Calculating water fowl abundance on the basis of banding returns , U.S.D.A.
Circ. 118 : 1-4
Linnaeus , C.1785 . Systema Naturae . 10th ed. , 1 : 824 pp. Holmiae .
Lockhead , J.A. 1950. Arthropoda : Xiphosura polyphemus in : Selected invertebrate types . pp.
360-381 . Brown , F.A. (ed) , New York , Wiley .
 152

Lockwood , S. 1870 . The horsefoot crab , Am. Nat . 4 : 257-274 , 3 pls .

Lowry , H. , Rosebrough , N.J. Farr , A.L. and Randall , R. J. 1951 . J. Biol. Chem. 193 : 265

Mackenzie , C. L.1962 . Tranportation of oyster drills by horseshoe crabs , Science 137 : 36-37

Maddrell , S. H. P. 1967 . Neurosecretion in insects . in : Beament , J.W.L. and J.E. Treherne (eds)
Insects and Physiology , Oliver & Boyd Ltd. Edinburgh and London , pp. 103-118

Mangrove Report (Annotated) , 1987 . A long term multidisciplinary research approach and report
on mangrove ecosystem of Sundarbans , Deptt. Mar. Sci. C.U. Calcutta .
Mangum , C. P. and Ricci , J. 1989 . The influence of temperature on oxygen uptake and transport in
the horseshoe crab Limulus polyphemus L ., J. Exp. Mar. Biol. Ecol. 129 (3) : 243-250
Manly , B. F. J. 1971 . A simulation study of Jolly's method for analysing capture-recapture data .
Biometrics 27 (2) : 415-424
Matsumoto , K.1954 . Neurosecretion in the thoracic ganglion of the crab , Eriocheir japonicus .
Biol. Bull. 106 : 60-68
Mc Mohan , T. A. 1973 . Size and shape in biology . Science 179 : 1201-1204
Mc Mohan , T. A. 1980 . Scaling physiological time ; Lectures on Mathematics in the Life
Sciences, 13 : 131-133
Mc Manus , J. J.1969. Osmotic relations in the horseshoe crab Limulus polyphemus , Am. Midl.
Nat. 81 : 569-573
Milne , L. J. and Milne , M. J. 1965.The crab that crawled out the past . G.Bell and Sons Ltd.London.
Miller , M. F. 1982. Limulicubichnus : A new Lchnogenus of Limulid resting traces . J. Paleontol .
52 (2) : 429-433
Miyata , T. , Tokunaga , F. Yoneya , T. , Yoshikawa , K. , Iwanaga , S. , Niwa , M . , Takao , T .
and Shimonishi , Y. 1989 . antimicrobial peptides , isolated from horseshoe crab hemocytes ,
tachyplesin II , and polyphemusins I and II : Chemical structure and biological activity .
J. Biochem . (Tokyo) . 106 (4) : 663-668
Mukherjee , K. N. 1983. Calcutta ; "Nature and problems of Neoreclamation in the Sundarban's"
Ind. J. Landscape syst . & Ecol. Studies . 6 : 1-19
Muller , O. F. 1785 . Entomostraca seu Insecta Testacea . 135 pp. Lipsiae et Hafniae .
Munford , R. S. , Hall , C. L. and Grim , L. 1984 . Detection of free endotoxin in cerebrospinal
fiuid by the Limulus laysate test . Infect. Immun . 45 (2) : 531-533
Mulaleedharan , D. K. and Prabhu , V. K. K. 1979 . Role of the median neurosecretory cells in the
secretion of protease and invertase in the red cotton bug , Dysderous cingulatus .
J. Insect. Physiol. 25 : 237-240
Muta , T. , Fujimoto , T. , Nakajima H. and Iwanaga , S. 1990 . Tachyplesins isolated from
hemocytes of South east Asian horseshoe crabs ( Carinoscorpius rotundicauda &
Tachypleus gigas) : Indentification of a new tachyplesin , tachyplesin III , and a processing
intermediate of its precursor . J. Biochem . (Tokyo) 108 (2) : 261-266
Nanda , D. K. , Bhowmik , L. K. and Choudhuri , P.S. 1984 . Histological study of neurosecretion
in the larval brain of Anomis sabulifera Guen (Noctuidae : Lepidoptera) . J. Zool.Soc.
India . 36 . (1&2) : 107-116
 153

Neff , J. M. and Giam , C. S. 1977 . Effects of Arochlor 1016 and Halowax 1099 on juvenile
horseshoe crabs Limulus polyphemus . in : Vernberg , F.J. et. al. , (eds.) Physiological
responses of marine biota to pollutants . Academic Press , New York , pp. 21-35.
Nishi , H. (ed) . 1975 . Kabutogami-Jiten (Encyclopedia of the Horseshoe Crabs) . Add. Rev. Ed. 221
pp. Private Publ. , Kasaoka , (in Japanese) .
Novak , V.J.A. 1975 . Insect hormones (2nd Edn). Chapman and Hall , London
Novitsky , T. J. 1982 . Limulus amoebocyte lysate : Live saving extract from horseshoe crab .J.
New England A quarium (A quasphere) 16 :26-31
Novitsky , T.J. 1984 . Discovery to commercializaion : The blood of the horseshoe crab . Oceanus .
27 (1) : 13-18
Page , C.H.1973. Loealization of LImulus polyphemus oxygen sensitivity . Biol. Bull.144 :383-390
Pal , S. G. , Sur , R. K. and Das , T. K. 1980 . Secience & Culture , 46 : 152
Panikkar , K . 1951 . Marine and estuarine fauna . In Chopra , B.N. (ed.) Handbook of Indian
fisheries . p. 24-33
Parameshwaran , R. 1956 . Neurosecretory cells of the central nervous system of the crab ,
Paratelphusa . Q. J. Micr. Soc. 97 :75
Parr , M. J. 1965. A population study of a colony imaginal Ischnura elegas (van der Linden)
(Odonata : Coenargriidae) at Dale , Pembrokeshire . Fld. Stud. 2 (2) : 237-282
Pattern , W . and Redenbaugh , W.A. 1899 . Studies on LimulusI .II. The nervous system of Limulus
polyphemus , with observations upon the general anatomy . J. Morph . 16 :91-200
Paul, A. K. 1986 . The development of accumulation forms and erosion in the coastal tracts of
Balasore Medinipur and 24-Parganas (Sundarbans) , West Bengal . Ind . J .Landscape
Syst. &Ecol. Studies , 9 (1) : 30-42
Pearse , A. S.1947. On the occurrence of actoconsortes on matine animals at Beaufort , N.C.
J. Parasitol . 33 : 453-458
Pearse , A. S.1949 . Observations on flat worms and nemarteans collected at Beaufort , NC .
Proc . US. Natl . Mus . 100 (3255) : 25-38
Pearson , F. C. and B Weary , M. 1980 . The Lilulus amoebocyte lysate test for endotoxin . Biosci .
30 : 461-464
Peters , R. H.1983.Ecological Implications of Body Size ,Cambr .Univ .Press ,Campridge ,U.K.p.329
Pocock , R . I. 1902 . The taxonomy of recent species of Limulus . Am. Mag. Nat. Hist. Sci.
9 (7) : 256-266
Pomerat , C. M.1933. Mating in Limulus polyphemus . Biol. Bull. 64 : 243-252
Pough , E. N.1980. The advantages of ectothermy for tetrapods . The Am. Nat. 115 : 92-112
Ramadan , A . A . and Matta , C. 1976. Studies on the central nervous system of the prawn. I. A
histological study for mapping the neurosecretory cells. 24 (2) : 148-154
Rama Rao , K. V. and Surya Rao , K. V. 1972. Studies on Indian King crabs (Arachnida : Xiphosura) .
Proc. Ind. Natl. Sci. Acad. 38B (3/4) : 206-211
Rao , D. V. and Bhagirathi , B. 1989. Carinoscorpius rotundicauda amoebocyte lysate for
detection of endotoxins : its preparation , stability , generitivity and comparison with
Limulus amoebocyte lysate . Ind. J. Exp. Biol. 27 (1) : 26-31
Riska , B.1981 . Morphological variation in the horseshoe crab , Limulus polyphemus.
Evolution, 35 :647-658
Robertson , J.D.1970 . Osmotic and ionic regulation in the horseshoe crab , Limulus polyphemus
(Linnaeus) . Biol. Bull. 141 : 157-183
 154

Roff , D. A.1973. On the accuracy of some mark-recapture estimators. Oecologia (Berl.) 12 :15-34
Roonwal , M. L.1944 . some observations on the breeding biology and on the swelling , weight ,
water-content and embryonic movements in the developing eggs of the Moluccan king crabs
Tachypleus gigas (Muller) (Arthropoda : Xiphosura). Proc.Ind. Acad.Sci. 20 :115-129
Ropes , J. W.1961 . Longevity in a horseshoe crab . Trans . Am. Fish. Soc. 90 : 79-80
Rose-Farhang , S. , Gadient , J. L. and Wasserman , G. S. 1990.Electrophsiological and behavioral
measures of seasonal and monthly variations in Limulus ventral eye sensitivity .
J. Interdiscip . Cycle . Res. 21 (2) : 97-118
Rudloe , A.1978 . Some ecologically significant aspects of the behaviour of the horseshoe crab ,
Limulus polyphemus . Ph.D. Thesis.The Floride State University ,Tallahassee, Floria .246pp
Rudloe , A.1979a. Limulus polyphemus : a review of the ecologically significant literature ,
P. 27-35 . In Cohen , E (ed) Biomedical Applications of the Horseshoe Crabs (Limulidae) ,
Alan R. Liss , Inc. New York.
Rudloe , A.1979b. Locomotor and light responses of larvae of the horseshoe crabs , Limulus
polyphemus (L.) Biol. Bull. 157 (3) : 494-505
Rudloe , A.1980 . The breeding behaviour and patterns of movement of horseshoe crab ,Limulus
polyphemus in the vicinity of breeding beaches of Aplachee Bay , Florida . Estuaries, 3 :
177-183
Rudloe , A.1981 . Aspects of the biology of juvenile horseshoe crabs , Limulus polyphemus .
Bull. Mar. Sci. 31 :125-133
Rudloe , A.1982 . Influence of man as an environmental threat to Limulus polyphemus. In
Bonaventura , J. et. al. , (eds.) Physiology and Biology of Horseshoe Crabs : Studies
on normal and environmentally stressed animals . Alan R. Liss , Inc . N. Y. U. S. A
Rudloe , A.1983 . The effect of heavy bleeding on mortality of the horseshoe crab , Limulus
polyphemus , in the natural enviroment J. Invert. Pathol. 42 :167-176
Rudloe , A. and Herrnkind , W. F. 1976 . Orientation of Limulus polyphemus in the vicinity of
breeding beaches . Mar. Behav. Physiol. 4 : 75-89
Rudloe , A. and Herrnkind , W. 1980. Orientation by horseshoe crabs , Limulus polyphemus , in a
wave tank . Mar. Behav. Physiol. 7 : 199-211
Rudloe , A. and Rudloe , J. 1981 . The changeless horseshoe crab. Natl. Geogr. 159 : 562-572.
Russel-Hunter , W. D. 1969 . A biology of higher invertebrates . Macmillan Company , New York .
Ryder , J. A. 1882 . Observations on the species of planarians on Limulus Am. Nat. 16 :48-51
Seber , G. A. F. 1965 . A note on the multiple-recapture census . Biometrika , 52 : 249-259
Seber , G . A. F. 1973 . The Estimation of Animal Abundance and Related Parameters . 506 pp. ,
Griffin , London .
Sekharan , K. V.1968 . Length-weight relationship in Sardinella albella (val.) and S. gibbosa
(Bleeker) . Ind .J. Fish. 15 : 166-174
Sekiguchi , K. 1960. Embryonic development of the horseshoe crab studied by vital staining :
Bull. Mar. Biol. St. Asamushi 10 :161-164
Sekiguchi , K. 1966 . Determination in the development of the horseshoe crab . Jap. J. Expt.
Morphol. 20 : 84-89
Sekiguchi , K. 1970a. On the embryonic moultings of the Japanese horseshoe crab , Tachypleus
tridentatus . Sci. Rep. 14 B : 121-128
Sekiguchi , K. 1970b. On the inner egg membrane of the horseshoe crab. Zool. Mag. 79 :115-118
(In Japanese).
 155

Sekiguchi , K 1973 . A normal plate of the development of the Japanese horseshoe crab Tachypleus
tridentatus . Rep. 15 B : 153-162
Sekiguchi , K. 1978. Surveys of horseshoe crabs in the Bay of Bengal and in the coast of South east
Asia. Jpn. Scientific Monthly , 31 :190-194 (In Japanese).
Sekiguchi , K. (ed.) 1988 . Biology of Horseshoe Crabs . Science House Co. , Ltd. Tokyo .pp. 440
Sekiguchi , K. , Ito , T. and Takasawa , J. 1976a . On a thin egg membrane formed inside the chorion
during the early embryonic development of the horseshoe crab. Acta Arachnol. 27(1) :
31-36
Sekiguchi , K. , Makioka , T. and Sato , T. 1976b. On the formation of the inner egg membrane in the
Japanese horseshoe crab ,Tachypleus tridenttatus . Zool. Mag. 85 (1) : 57-64
Sekiguchi , K. and Nakamura , K.1979 . Ecology of the extamt horseshoe crabs in Cohen , E. et.al.
(eds.) . Biomedical Applications of the Horseshoe Crab (Limulidae) , Alan R. Liss. Inc. N.Y.
pp. 37-45
Sekiguchi , K. and Nakamura , K. 1980 . Sympatric distribution of three species of Asian
horseshoe crabs. Proc. Jap. Soc. Syst. Zool. 18 :1-4
Sekiguchi , K. , Nakamura , K. , Sen , T.K. and Sugita , H. 1976 . Morphological variation and
distribution of a horseshoe crab , Tachypleus gigas , from the Bay of Bengal and
the Gulf of Siam . Ibid . 12 :13-20
Sekiguchi, K. ,Nakamura, K. and Seshimo, H.1978.Morphological variation of a horseshoe
crab, Carcinoscorpius rotundicauda , from the Bay of Bengal and the Gulf of Siam.
Ibid. 15 : 24-30
Sekiguchi , K. , Nishiwaki , S. , Makioka , T , Srithunya , S. , Machjajib , S. Nakamura ,K. and
Yamasaki , T. 1977. A study on the egg laying habits of the horseshoe crabs , Tachypleus
gigas and Carcinoscorpius rotundicauda , in Chonburi area of Thailand. Ibid. 13 : 39-45
Sekiguchi , K. , Seshimo , H. and Sugita , H.1988 . Post embryonic development of the horseshoe
crab . Biol. Bull. 174 : 337-345
Sekiguchi , K. and Sugita , H. 1980. Systematics and hybridization in the four living species of
horseshoe crabs. Evolution 34 (4) : 712-718
Sekiguchi , K. Yamamichi ,Y. and Costlow , J. D. Jr. 1982 . Horseshoe crab developmental studies I.
Normal embryonic development of Limulus polyphemus compared with Tachypleus
tridentatus in Bonaventura , J. et.al. , (eds.). Physiology and Biology of Horseshoe
Crabs: Studies on normal and environmentally stressed animals , pp , 53-73 . Alan R. Liss.
Inc. New York.
Selander , R. K. and Yang. S.Y. 1970. Horseshoe crab lactate dehydrogenase : Evidence for dimeric
structure . Science . 169 179-181
Selander , R.K. , Yang, S. Y. , Lewontin , R. C. and Johnson , W. E. 1970. Genetic variation in the
horseshoe crab (Limulus polyphemus) , a phylogentic " relic " . Evolution 24 : 402-414
Scharrer , B.1941. Neurosecretion . IV Localization of neurosecretory cells in the central nervous
system of Limulus . Biol. Bull. 81 : 96-104
Scharrer , E. and Scharrer , B. 1954. Neurosecretion V . Mollendorffs Hanb. D. mikr. Anat. d.
Menschen. 6 (5) : 953-1066
Schlottke , E. 1934a. Histologische Beobachtungen am Darmkanal von Limulus und vergleich
vergleich seinen Zwischengewebes mit dem von Spinnen . Zeitsch. Mikrosk. Anat.
Forsch. 34 . 57-70
 156

Schlottke, E.1934b.Unterschiede in der Entwicklung des phagozytierenden und des resorbier-

enden Datmepitteles . Biol. Zbl. 54 :51-64
Schlottke , E. 1935. Biologische , physiologische , und histologische Untersuchung uber die
Verdauung von Limulus . Z . vergl. Physiol. 22 : 359-413
Seligman , A. M. and Nochlas , M . M. 1963. Methods in Enzymatic Analysis , Bergmeyer , H. (ed.).
Acad. Press , N. Y. and London , p. 776
Sellenschlo , T. 1983. Encounter with Ziphosura . New Entomol . Nachr. O (5) : 9-12
Sewell , R. B. S.1912. Capture of Limulus on the surface . Rec. Ind. Mus. 7 : 87-88
Sheppard , P. M. , Macdonald , W. W. , Tonn , R. J. and Grab , B. 1969. The dynamics of an adult
population of Aedes aegypti in relation to dengue haemorrhagic fever in Bankok .
J. Anim . Ecol. 38 : 661-702
Shipley , A. E. 1909 . introduction to arachnida and Xiphosura in Harmer , S.F. and A. E. Shipley
(eds.). The Cambr. Nat. Hist. col. 4 pp. 253-279 , Mac Millan & Co. , London.
Shishikura , F. , Nakamura , S. , Jakahashi , K. 1982 . Horseshoe crab phylogeny based on amino
acid sequence of fibrino-peptide like C. J. Exp. Biol. 223 (1) : 89-92
Shoji , K. 1927.On the egg - spawning of the horseshoe crab. Zool. Mag.Tokyo , 39 :55-59(in Japanese)
Shoji , K. 1929a. Laevae of Limulus longispinus and its metamarphosis. Ibid. 41 :28-36 (in Japanese)
Shoji , K. 1929b. Kabuto-gani (The japanese horseshoe crab) in Okada , Y. (ed.), Collected Atlas of
Anatomy of Animals , part 3 , pp. 109-160 , Kyoritsu - sha , Tokyo ( in Japanese)
Shoji , K. 1932 . Morphology and ecology of Limulus longispinus . Fukuoka Hakubutsu - gaku
Zasshi , 1 (1) : 28-37 (in Japanese)
Shuster , C. N. Jr. 1950. Observations on the natural history of the American horseshoe crab ,
Limulus polyphemus . 3rd Rept. , Investigations of Methods of Improving the Shellfish
Resources of massachusetts , Woods Hole Oceanographic Inst. Contr. No. 564 :18-23
Shuster , C. N. Jr. 1948. On the gross anatomy and histology of the alimentary tract in early
developmental states of Xiphosura ( = Limulus) polyphemus Linn. M.Sci. Thesis (Unpubl.)
(Rutgers Univ . N.J. 50 pp.
Shuster , C. N. Jr. 1953. Odyssey of the horseshoe "crab". Audobon Mag. 55 (4) : 162-163
Shuster , C. N. Jr. 1954. A horseshoe "crab" grow up. Ward's Nat. Sci. Bull. 28 (1) : 1-6
Shuster , C. N. Jr. 1955. On morphometric and serological relationships within the Limulidae , with
particular reference to Limulus polyphemus (L) Diss. Abstr. 18 (2) : 371-372 .
Shuster , C. N. Jr. 1957. Xiphosura (with special reference to (Limulus polyphemus) .
In Hedgpeth , J. W. (ed.), Treatise of Marine Ecology & Paleoecology , vol. 1 Geol. Soc.
Am. Mem. 67 (1) : 1171-1174
Shuster , C. N. Jr. 1958. "Study thesis" - the story of the horseshoe crab . Staff Reporter ,
Wilmington Public Schools , Delaware , 10 : 4-5
Shuster , C. N. Jr. 1960a. Horses "Crabs" in former years , during the month of May , these animals
dominate Delaware Bay shores . Estuarine Bulletin 5 (2) : 1-9
Shuster , C. N. Jr. 1960b. Xiphosurida , in Eneyclopedia of Sci &tech. Mc Graw Hill Book Co. Inc .
14 : 563-567 (revised 1971 and 1982 = Xiphosurida 14 : 766-770)
Shuster , C. N. Jr. 1962. Serological correspondence among horseshoe "crabs" (limulidae).
Zoologica , 47 : 1-8 , + pl. 1.
Shuster , C. N. Jr. 1978. The circulatory system and blood of the horseshoe crab. U . S. Dept. of
Energy , FERC , Washington , D.C. pp. 1-63
 157

Shuster , C. N. Jr. 1979. Distribution of the American horseshoe "crab" , Limulus polyphemus (L.)
in Cohen , E. et.al. , (eds.) Biomedical Applications of the Horseshoe Crab (Limulidae) .
p. 3-26 , Alan R. Liss. N. Y.
Shuster , C. N. Jr. 1982. A pictorial review of the natural history and ecology of the horseshoe crab ,
Limulus polyphemus , with reference to other Limilidae . p. 1-52 , in Bonaventura ,J.
et. al. , (eds.) Physiology and Biology of Horseshoe Crabs . Alan R. Liss. Inc. New York .
Shuster , C. N. Jr. 1988. Research on Xiphosura : In Retrospect / prospect , presented at the
Scientific Conference on Limulus polyphemus. Marine Biological , Inc. New Jersey .
p. 1-13 . + Appendix 1-8 .
Shuster , C. N. Jr. and Horrell , H. C. 1966 LImulus exoskeletion as a teaching aid. Turtox News ,
44 (1) : 40-41
Shuster , C. N. Jr. and Botton ,M. L. 1985 . A contribution to the population biology of Horseshoe
crabs , Limulus polyphemus (L.) in Delaware Bay . Estuaries 8 (4) : 363-372
Shuster , C. N. Jr. and Langrall , C. H. P. and Shevock R.C. 1992 . Biometric analysis of sexual
dimorphism in mated pairs of Limuslus polyphemus from Delaware Bay . Adv. Biosci.
(in press)
Sluys , R.1984. A new marine triclad ectoparasite on Malayasian and Indonesian horseshoe crabs .
Bijdr. Dierkd . 53 (2) : 218-226
Smedley , N.1929 . Malayasian King crab . Bull. Raffles Mus. 2 :73
Smith , O. R.1953 . Notes on the ability of the horseshoe crab , Limulus polyphemus , to locate
soft - shell clams , Mya arenaria . Ecology 34 : 636-637
Smith , O. R. and Chin , E. 1951 . The effects of predation on soft clams , Mya arenaria . Natl.
Shellfish Assoc . Conv . Adress , p. 37-44
Smith , O. R. , Baptist , J.P. and E. Chin ,1955 . Experimental farming on the soft-shell clam ,
Mya arenaria , in Massachusetts , Comm . Fish . Rev. 17 (6) : 1-16
Smock , L. A.1980. Relationship between size and biomass of a quatic insects . Freshwater
Biology, 10 : 375-383
Snedecor , W. H. and Cochran , W. H. 1967 . Statistical Methods . The lowa State Univ. Press , USA
Snodgrass , R. E.1952 . A textbook of Arthropod Anatomy . X + 363 pp. Ithaca .
Sokoloff , A. 1978 . Observations on populations of the horseshoe crab Limulus ( = Xiphosura
polyphemus) . Res. Popul. Ecol. 19 : 226-236
Southwood , T. R. E. 1978 . Ecological Methods : with particular reference to the study of insect
population . 2nd Edn. ELBS + Chapman and Hall . , London , U.K.
Srimal , S. , Miyata , T. ,Kawabata , S. , Miyata , T. and Iwanaga , S. 1985. The complete
aminoacid sequence of coagulogen irolated from South east Asian horseshoe crab ,
Carcinoscorpius rotundicauda . J. Biochem . 98 : 305-318
Stomer , L.1952.Phylogency and taxonomy of fossile horseshoe crabs . J. palentol. 26 (4): 630-639

Stomer , L.1955. Merostomata . In Moore , R. C. (ed.) . Treatise on Invertebrate Paleontology , part P.

pp. 4-41 Geol. Soc. Am. Univ. Kansas press , lawrence .
Sugita , H. , Murakami , K. and Sekiguchi , K. 1985 . The effect of salinity on embryonic
development of the horseshoe crabs . Acta Arachnol . 34 (1) : 1-10
Sugita , H. and Sekiguchi , K. 1979 . Protein components in the previtellini fluid of the embryo of
the horseshoe crab , Tachypleus tridentatus . Develop. Biol. 73 : 183-192
 158
Sugita , H. and Sekiguchi , K. 1982 Horseshoe crab developmental studies II . Physiological
adaptation of horseshoe crab embryos to the environment during embryonic development .
in Bonaventura , J. et. al. , (eds.) Physiology and Biology of Horseshoe Crabs.
pp.75 - 82 , Alan R. Liss. Inc. New York.
Sugita , H. and Sekiguchi , K. 1983. The developmental appearance of paternal forms of lactate
dehydrogenase and males dehydrogenase in hydrid horseshoe crabs . Biol. Bull. 165 : 436-443
Sugita , H. and Sekiguchi , K. 1984. Lactate dehydrogenase of the horseshoe crabs and their
hybirds . Zool. Sci. 1 :421-426
Sugita , H. and Sekiguchi , K. , Shishikura , F. and Yamamichi , Y. 1982 . An evolutionary aspect to
horseshoe crabs based on developmental capacity of the interspecific hybirds . Proc. Jap.
Soc. Syst. Zool. 22 :1-6 (in Japanese) .
Tanimoto , S. and Kondo , A. 1982 . Histochemical localization of some enzymes in mid-gut of
Japanese horseshoe crab , Tachypleus tridentatus . Zool. Mag. (Tokyo) . 91 (2): 135-139
Teale , E. W. 1957. The oldest migration Nat. Hist. 66 : 364-369
Thiemmedh , J .1965 . Poisonous king crab ( Caecinoscorpius rotundicauda) . Notes Fac. Fish.
Kasetsurt Unit (Bankok) 2 : 3-4
Thomsen , E. and Moller , I.B. 1959. Neurosecretion and intestinal protease activity in an insect,
Calliphora erythrocephala . Nature 183 : 1401-1402
Thys , J. P. 1984 . Diagnosis and treatment of bacterial meningitis . Resuscitation 11 (3/4):243-248
Turner , H. J. Jr . , Ayers , J. C. Wheeler , C. L. 1948. The horseshoe crab and boring snail as factors
limiting the abundance of the soft shell clam . Woods Hole Ocean , Inst. Contr. 462:43-45
Van der Hoeven, J., 1983. Recherches sur 1" Histoire naturelle et 1" Anatomic des Limules. 48 pp.7
pls. Luchtmans, Leyde.
Verrill, A.E. 1983. marineplanarians of New England. Trans CT. Acadl. 8 : 459-520.
Verrill, A.E. 1985. Supplement to the marine nemarteans and planarians of New England. Trans.
CT. Acad. 9 : 141-152.
Vijaya Rao, D. and Bhagirathi, B. 1989. Crcinoscorpius rotundicauda amoebocyte lysate
detection of endotoxins - its preparation, stability, sensitivity and comparsion with
Limulus amoebocyte lysate. Ind. J. Exp. Bion. 27 : 26-31.
Vosatka, E. D. 1970. Observations on the swimming, righting and burrowing movements of young
horseshoe crabs, Limulus polyhemus. Ohio. J. Sci. 70 (5) : 276-283.
Waterman, T.H.1953. Xiphosura from Xoung - Ha. Am. Scint. 41 : 292-302.
Waterman, T.H.1954. Relative growth and the compound eye in Xiphosura. J. Morphol. 95:125-128.
Waterman. T.H.1958. On the doubtful validity of Tachypleus hoeveni Pocock, an Indonseian
horsesoe crab (Xiphosure). Positilla Yale peabody Mus. 36 : 1-17.
Wheeler, W.M.1894. Syncoelidium pellucidum, a new marine triclad. J. Mrophol : 167-194.
Willemoes-Suhn,1883. On a crustacean larva at one time supposed to be larvae of Q. J. R. Von.
Microscopic. Sci. 23 : 145-150.
Wyse, G.A.1971.Receptor organization and function in Limulus Chelae.Z.Vergl.Physion.73:249-273.
Yamamichi,Y. and Sekiguchi, K.1974.Embryo and organ cultures of the horseshoe crab,
Thchypleus tridentatus. Dev. Growth. Diff. 16 (4) : 295-304.
Yamamichi,Y., Sugita, H., and Sekiguchi, K. 1983. Morphological characterization of first instar
larva of Asian horseshoe crabs and their hybrids. Dev. Growth Diff. 25(3):271-280.
Yamasaki, T., Makioka, T. and Saito. J.1988. "Morphology" in Sekiguchi, K. (ed.) Bioligy of
Horseshoe Crabs. Science House Co., Ltd. Tokyo, pp. 69-132.
Zhao, Y. 1983. Effects of Limulus lysate against lethal action of endotoxions. Tianjin Med. J.
11 (10) : 607-609.
End of T he T hesis Paper
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