Indian Journal of Geo-Marine Sciences

Vol. 40(4), August 2011, pp. 516-521

Sex-ratio, maturation and spawning of the Indian ilisha, ilisha melastoma

(clupeiformes: pristigasteridae) in coastal waters of Pakistan
(northern Arabian Sea)
Khalid Mahmood , Zarrien Ayub* & Ghazala Siddiqui
Centre of Excellence in Marine Biology, University of Karachi, Karachi-75270, Pakistan
*
[E-mail address: zarrienayub@yahoo.com]
Received 22 March 2010; revised 5 December 2010
Present study showed that the over-all sex-ratio deviated in favour of females in Ilisha melastoma. However, the sexratio was approximately 1:1 in certain months while in others it was in favor of females. Similarly the sex-ratio deviated
from 1:1 ratio in different size-classes. Minimum size at which the specimens of I. melastoma get matured is 14.9 cm in total
length (11.5 cm in SL). Four maturation stages of ovaries, namely, immature, ripening, ripe and spent/recovering were
recognized in I. melastoma. Two peaks of spawning were observed in this species, the major peak was from February to
May and the minor peak appeared in August to November.
[Keywords: Maturation, spawning, Indian ilisha, Pakistan]

Introduction
Species of the genus Ilisha are distributed in the
Indo-Pacific region, i.e., Japan, China, Hong Kong,
Singapore, Java, Borneo, Sumatra, Burma, India,
Pakistan, Persian Gulf and Africa. According to
Whitehead1 genus Ilisha have a circumtropical
distribution in estuaries and coastal waters, with one
species in the tropical east Atlantic, two in South
America and eleven in tropical Asia. Genus Ilisha is
important in artisanal fisheries throughout the tropics
of South and Southeast Asia, however little is known
about the species belonging to genus Ilisha2-4.
Two species of genus Ilisha have been reported
from the coastal waters of Pakistan, I. melastoma and
I. megaloptera5. Information on the reproductive
biology of a species, such as length and age at first
maturity, spawning season and fecundity are
important for effectively managing the fisheries for
that species6. In the present study, the sex-ratio,
maturation and spawning of Ilisha melastoma (Bloch
and Schneider, 1801) was studied based on
macroscopic and microscopic examination of gonad.
Materials and Methods
Samples of Ilisha melastoma were procured
monthly from the landings at Karachi Fish Harbour
_________________

For Correspondence

during the period March 2004 to February 2005 and a

total of 725 specimens of I. melastoma were
examined. Total length (cm) and weight (g) of each
fish was recorded. Fish was dissected from the
abdominal region and the sex and gonadal maturity of
fish was determined by visual examination of gonads
(Table 3). For histological study, every month the
ovaries of 20-25 fishes were fixed in Davidsons
Fluid for 48 hours. After fixation the ovaries were
transferred to 70% alcohol. Process of dehydration,
infiltration and embedding was followed as
mentioned in Humason7. Embedded ovaries were
sectioned at 7 m and then stained with Delafields
Haematoxylin and counterstained with Eosin.
For fecundity estimates, 33 ripe ovaries were
removed and weighed accurately by using electrical
balance to nearest 0.01 g. From each ovary three
subsamples from anterior, middle and posterior region
of the ovary, weighing 0.1 g were preserved in 10%
formalin for 5 days. Each and every ripe oocyte in the
subsample was counted under the binocular
microscope and the fecundity was estimated
according to Hunter et al.8.
F = W/w N
where F = Fecundity; W = Total ovary weight;
w = subsample weight; N = no of oocytes in the
subsample.

KHALID et al.: SEX RATIO, MATURATION AND SPAWNING OF THE INDIAN ILISHA MELASTOMA

The oocyte diameter was measured on the

histological sections with the help of an ocular
micrometer. The diameter of oocyte was taken at one
axis running parallel to the slide and the widest part of
the oocyte was measured.
Results and Discussion
Sex-ratios

Chi square (2) was used to test whether the sexratios observed were significantly different from the
expected sex-ratio of 1:1. The overall sex-ratio of
I. melastoma was significantly in favour of females
(X2 = 25.98; P < 0.001) (Table 1). Month-wise
distribution of sexes fluctuated significantly in favour
of females in May, June, August, and February while
in November it showed the preponderance of males
(X2 = 9.68; P = < 0.01). In the remaining months it
followed 1:1 sex-ratio. In I. africana respectively
from Nigeria and Ghana, the over-all sex-ratio did not
differ significantly from the expected 1:1 ratio9-10,
however, the occurrence of more females in certain
months was related to the peak of their spawning
periods9. Presence of more females of I. melastoma in
the present study in February, March, May, June, and
August coincides with the peak of their spawning.
Table 1 Chi square test of sex-ratios of I. melastoma in
various months during the period from March 2004 to February
2005 (*P<0.05; **P<0.01; ***P<,0.001).
Months

Total

March
April
May
June
July
August
September
October
November
December
January
February
Total

23
54
102
77
72
75
60
70
50
61
45
36
725

Males Females Unsexed

11
23
39
11
27
21
20
32
36
29
24
1
274

12
31
63
28
41
54
39
37
14
32
21
35
407

38
4
1
1
44

x2
0.04
1.19
5.65*
7.41**
2.88
14.52***
6.12*
0.36
9.68**
0.15
0.20
32.11***
25.98***

517

Deviation in sex-ratio could be due to partial

segregation of mature forms through habitat
preferences11 due to migration12 or behavioral
differences between sexes13 thus rendering one sex to
be more easily caught than another.
The 1:1 ratio of I. melastoma was found to be
disturbed in various size-classes (Table 2). Males
showed preponderance in the size-class 14.0-15.9 cm
(X2 = 6.26; P < 0.05). Size-class 16.0-17.9 cm
followed the ratio 1:1, while in the remaining sizeclasses the 1:1 ratio was in favour of females
(Table 2). Fish >24.0 cm showed the males and
females approximately in equal numbers. Differences
in the size-specific sex-ratio in fish have been related
to sexual differences in growth, mortality or energetic
cost of reproduction14-15.
Macroscopic examination of gonads

The sex of I. melastoma was determined on the

visual examination of the gonads and four maturation
stages of male and female gonad of I. melastoma were
recognized, that is, immature, ripening, ripe and
spent/recovering whose characteristics are given in
Table 3. However, the sex of 44 specimens of
I. melastoma could not be identified on macroscopic
examination and the sex in these individuals remained
indistinct even on histological examination of the
gonad. Unsexed individuals of I. melastoma may be
the spent condition of the gonad as has been reported
Table 2 Chi square test of sex-ratios in different size-classes
of I. melastoma obtained from the landings of Karachi Fish
Harbour (*P<0.05; **P<0.01; ***P<,0.001)
Size classes
(cm)

Total

Males

Females

Unsexed

X2

14.0-15.9
16.0-17.9
18.0-19.9
20.0-21.9
22.0-23.9
24.0-25.9
26.0-27.9

31
215
291
145
25
13
5

20
87
120
33
7
3
4

7
95
166
110
18
10
1

4
33
5
2
-

6.26*
0.35
7.40**
41.46***
4.84*
3.77
1.80

Table 3 Macroscopic description of the maturity stages of the gonads of I. melastoma

Stages
Females
Males
Stage 1 (Immature)
Translucent or whitish and thread-like, no White and thread-like, histological examination revealed
oocytes visible, histological examination the sex
revealed the sex
Stage 2 (Ripening)
Pinkish with blood capillaries, eggs appear as Whitish or creamish, size of gonad increased and can be
whitish granules, size of gonad further increased recognized as male gonad as milt extruded on pressure
Stage 3 or Ripe
Pinkish or reddish, eggs distinctly visible, gonad Whitish with pink vascularization, gonad occupy of
occupy of the body cavity
the body cavity, milt running on pressure
Stage 4 or
Pinkish or reddish, eggs opaque, gonad shrinks
Whitish, flaccid, gonad seems empty
Spent/Recovering

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INDIAN J. MAR. SCI., VOL. 40, NO. 4, AUGUST 2011

Fig. 1 Photomicrographs showing the developmental stages of the ovary in I. melastoma. A. Section of an immature ovary containing
chromatin nucleolar and perinucleolar oocytes. B. Section of a ripening ovary showing perinucleolar oocytes and partially yolked
oocytes. C. Section of a ripe ovary showing advanced yolked oocytes. D. Section of a spent/recovering ovary showing atretic oocytesb
and perinucleolar oocytes. CN, chromatin nucleolar oocytes; PO, perinucleolar oocytes; PYO, partially yolked oocytes; AYO, advanced
yolked oocytes; AO, atretic oocytes, Scale = 100 m.

by West16 that such gonads were probably spent

ovary in which the egg production has ceased for the
current season and therefore, present problems in
identifying.
Histological staging of female gonads

Four arbitrary maturation stages of ovaries were

recognized in I. melastoma on the basis of histological
examination (Table 4 and Figure 1).

Size at first maturity of females

Smallest female of I. melastoma with ripe ovary

measured 14.9 cm in total length (11.5 cm in SL). The
18.0-19.9 cm size-class showed the greatest number
of I. melastoma with ripe ovaries. Blaber et al.4
reported that I. melastoma matured at about 100 mm
SL and this size is smaller than the size (11.5 cm in
SL) obtained in the present study. Size of first sexual
maturity in females of some other Ilisha species

KHALID et al.: SEX RATIO, MATURATION AND SPAWNING OF THE INDIAN ILISHA MELASTOMA

519

Table 4 Microscopic description of the maturity stages of

the ovaries of I. melastoma.
Stages

Ovaries

Stage 1 (Immature)

Oogonia, chromatin nucleolar

and perinucleolar oocytes are
present. The perinucleolar
oocytes had a large nucleus.
Oocytes
diameter
ranges
between 25-60 m.

Stage 2 (Ripening)

Partially yolked oocytes and

perinucleolar oocytes present.
The partially yolk ooyctes
contained yolk granules and
few yolk globules in their
cytoplasm.
Oocytes
diameter
ranges
between 60-200 m.

Stage 3 (Ripe)

Advanced yolked oocytes are

abundantly
present.
The
cytoplasm of the oocytes are
filled with more yolk globules
and few yolk granules.
Oocytes
diameter
ranges
between 200-300 m.

Stage 4 (Spent/Recovering)

Ovary containing perinucleolar

oocytes. Few atretic oocytes
and post-ovulatory follicles can
also be seen.

reported are for I. africana from Nigeria had

TL 11.7 cm (ref.9); I. kampeni matured at about 100
mm SL, I. megaloptera, I. elongata and
I. pristigastroides matured between 100 and 200 mm
SL while I. filigera matured at about 400 mm from
Sarawak, Malaysia4. While, Yankson and Azumah10
reported that the smallest mature female of I. africana
from Ghana had TL 13.2 cm.
Spawning season based on macroscopic
examination of gonads
Males

On the basis of the macroscopic examination of the

male gonads, it was found that ripe males of
I. melastoma were present throughout the year with
peak during September to November period (Figure 2).
Females

On the basis of the macroscopic examination of the

female gonads, it was found that ripe females of
I. melastoma were present during the period from
February to May and from August to November
(Figure 2).
Spawning season based on histological staging of
female gonads

Fig. 2Monthly percentages of ripe males and females

(based on macroscopic examination of gonads) of I. melastoma
from March 2004 to February 2005.

The spawning season is based on the presence of

females with ripe ovaries. With the exception of
certain months the ripe females were found
throughout the year (Figure 3). However, two peaks
of spawning were observed in I. melastoma, the major
peak from February to May and the minor peak from
August to November. Marcus and Kusemuje9 reported
that the ripe and ripe running females were caught in
all months indicating that the breeding occurred
throughout the year in I. africana off the Lagos coast,
Nigeria. However, the peak spawning was observed
between May and December. Yankson and Azumah10
had reported the major spawning season during
January to May in I. africana off Cape Coast, Ghana.
Previous studies have indicated that many clupeids
bred throughout the year17-18.
In the present study the spawning season of
I. melastoma based on the macroscopic examination
of the gonads was in agreement with the spawning
season, from February to May and from August to
November, based on the histological examination of
the gonads. However, the use of histology in maturity
studies has become more and more widespread as it is
more consistent and reliable19.
Fecundity

Egg counts were made of females ranging in size

between 17.5 to 25.7 cm in total length and
48 to 123 g in weight. The fecundity ranged from
4392 to 32756 eggs.
Relationship between fecundity and fish total
length (TL) and fecundity and fish weight (W) was
estimated and described by the equation:
F = aXb

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INDIAN J. MAR. SCI., VOL. 40, NO. 4, AUGUST 2011

Fig. 3Monthly percentages of developmental stages of I. melastoma ovaries (based on microscopic examination) from
March 2004 to February 2005. 1 = Immature stage; 2 = Ripening stage; 3 = Ripe stage; 4 = Spent/recovering stage.

where F = Fecundity, X = an independent variable

a and b were estimated by using linear regression
as shown in the equations:
F = 0.011 TL4.675
F = 995.4 W0.623
There was no significant relationship between
fecundity and total length (r2 = 0.298; P > 0.05) and
fecundity and body weight (r2 = 0.030; P >0.05).
In the present study the fecundity in I. melastoma
ranged from 4392 to 32756 eggs. The fecundity
reported in the present study is greater than that of
Marcus and Kusemuje9 and Yankson and Azumah10 in
I. africana which ranged from 2098 to 11687 and

1013 to 10600 eggs, respectively. In the present study

no significant relationship was found between
fecundity and fish length or weight. According to
Blaber et al.4 there is a positive relationship between
fecundity and fish length in species of Ilisha. King20
had also conformed to this relationship in I. africana.
According to Marcus and Kusemuje9 fecundity in
I. africana increased with fish length and weight and
the increment of fecundity relative to the length and
weight of fish were nearly isometric.
The result of this study of I. melastoma will
provide valuable information to the fisheries
biologists and fisheries management authorities as
this species has wide distribution from India to
Indonesia and to Taiwan Islands1.

KHALID et al.: SEX RATIO, MATURATION AND SPAWNING OF THE INDIAN ILISHA MELASTOMA

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