WATER, BATII STUDIES OX' MAMMALS FROM SEVERAL ECOI,OGIOAL AIi,EAS

II. L. RIEDESEL and J. T YELVIRTOI\

Biology Depart'ment, The {Jniversity of New Mexico, Albuquerque, New Mexico, U.S.A.

Reprinted lrom BIOI{ETEOROLOGY

Proceedirt gs

II.
C

o| the T hird. Internati,onal B'iometeorological held at Pau, S. Irance, 7-7 Beptember 7963
PETI,CA-\ION PRESS

ongress

OXFOIiD LONDON EDINBUR,GTI NEW YORK TORONTO PAR,IS FR,ANI(FLTR,T

1966

WATER, BATII STUDIES OF MAMMALS FROM SEVERAL ECOLOGICAL AREAS*

M. L. RIEDEStsL anrl J. T. YELYERTON
Biology Deparfment, The University of Nerv Mexico, Albuquerque, New Mexico, U.S.A.
Alrsl,raet - Waterbath (25" C) experiments onC. Iateralis C. spilosoma M. auratus .aneJ. ord,i' demonstrate differences in the capacities of these animals to tolerate a severe cold stress. The laleralis and spilosomo, Lrad a, greater capacitv for shivering. Tbe ktte.ralis had a greal,er al-.ility to limit heat loss by peripheral vasoconstriction. The principal method of combating cold available to ordi ancl auratus rvas preferential cooling of peripheral

D.

tissues.

Zusammentassung - C.lateralis C. spilosorna M. auratus und D. ordi zeigten unt,erschiedliclie Kiilteempfindlichkeiten im Wasserbadexperiment, (25' C). Laterolis rnd spilosoma habten grbssere Kapazitiit fiir Muskelzittern. Lateralis verhinderte den \4'5rmever1ust,

am besten durch periphere Gefd,ssverengung. Die I(dlteabwehr in ordi und auratus beruhte hauptseichlich auf der schnelleren Abkiihlung der d,usseren Gervebe. R6sum6 - C.l'ateral'is C. sptilosoma M. auratus el D. ordi d6montrent diff6rentes sensibilit6s au froid dans I'exp6rience i, l'eau (25' C). Laleralis el spilosoma d6montrent une

plus grande capacit6 de frisson. Lateral,is limitait la perte de chaleur par striction des vaisseaux sanguins peripheriques. La principale m6thode de combattre le froid repose avant tout sur une possibilit6 plus rapide de rafraichissement des tissus extdrieurs.

Bnnevron patterns and insulative values of fur are recognized as being primarv factors in determining tolerance of anima,ls to thermal stress (Burton a,nd Edholm, 1955; frving and Krog, I954; Prosser and Brown, l96l). Few studies have explored physiological differences which may be important in determining the transfer or conseryation of body heat. Physiological capacity may be of great importance in determining the survival of a given species at extremes in the thermal environment, for instance: Irhen the limits of protection by insulation and behavior ha,ve been
surpassed, physiological capacity would determine survival. Regarding physiological capacity there is evidence of species differences in temperature regulation, zone of thermal neutrality, tolerance for hypothermia and cooling of peripheral tissues (Hannon and Viereck, 1955; Kayser, 196l; Prosser and Brown, 1g6f). Additional information is needed in these areas, particularly with respect tophysiological capacity of animals from various ecological groups. The v'ater bath is a

* This study

was supported in part by NSF Grant GB216 and a NSF fellowship.

s90

Water Bath Studies of Mammals

891

useful tool for describing transfer and conservation of body heat in response to thermal stress because environmental factors, such as radiant heat, air velocity, fluctuating air t'emperature and humidity are not involved. (Wilber, 1957).

MATERIALS AND MNTHODS

The animals studied include three hibernators from different ecological areas and a non-hibernator. Ci,tellus spilosoma rnclrg,inatus inhabit warm regions (mean air t'emperature - summer, 24' C; winter, 3" C) at elevations of 1500 to 2000 m. Ci,tellus lateral'is lateralis inhabit cool regions (mean air temperature - summer, 13" C; winter, -7" C) atelevations of 2600 to 4300 m. Mesocreci,tus auratus, Asian ancestory, were purchased from a pet shop. The fourth species, Dipotlomys ord,'i, are nocturnal non-hibernators and inhabit the same areas as spilosoma in New
Mexico.

A1l animals had been in the laboratory for I to 12 months prior to the studies. Periodic body weight measurements to the nearest 0.5 g indicated they were in good health: mean body weight 231 g (7.0f s.n.) lateralis; 132 g (4.58 s.n.; spilosoma; 1f8 g (2.99 s.n.) auratus; 68 g (3.68 s.n.) ordi. The swim chamber (33X33X25.5 cm) was constructed from 1.2 cm lucite. A rubber gasket fitted to a groove on the top edge of the chamber insured an airtight seal when the lid was fastened to the chamber with adjustable vise grip clamps. One side of the chamber was fitted with a drain outlet, while the lid was fitted with four outlets for temperature probes and an oxygen source. A wire-meshbag athached to the underside of the lid filled with soda lime provided absorption of carbon dioxide. A heating element of 110 v, 300 w resistance wire in a coiled aluminum tube was inserted through the drain outlet. Cooling was effected by immersion of the swim chamber in a I00 I water tank which provided constant temperatures t0.4o C. Metabolic rate determinat'ions were made by measuring oxygen consumption with a spirometer sensitiye to 0.6 cm3. All rectal and water temperature me&surements were made with thermistor probes. Three series of water immersion experiments were conducted : (1 ) free swimming, (2) confined, and (3) dead animals for a total of 96 water bath experiments on 14 lateralis, 10 spilosoma, 8 ordi and 4 auratus. Repetitive exposure to cold water did not effect acclimatization as reported by Adolph and Richmond (1956). The protocol for the free swimming experiments of 30 min duration was as follor'vs: weigh animals, measure rectal tempera,ture, place in water at time zeto, and. ai l0 min intervals the a,nimals were removed from the water for I - 2 rnin for rectal temperature measurements. Metabolic rate me&surements were made during the second 10 min swim period. The series on confined and dead animals involved placing the animals in a cone of one centimeter mesh wire and immersing to the head in water. Rectal temperature determinations of dead animals were made at two minute intervals for the first l0 min and at 5 min intervals for the last 20 min.

892

M. L. Rrnnnsrl aND J. T. Ynr-vnnroN

Short term experiments of 2, 4, 6, 8, and l0 min duration were conducted on swimming animals. fn this series the animal was removed from the bath at the specified times: 2, 4, 6,8, ancl l0 min, and rectal temperature w&s recorded. Three animals of each species were tested for each time interval and a given a,nimal rvas subjected to no more than one swim per day. Several assumptions have been made to facilitate interpretation of the data: (f ) Heat loss from the head which was above the water in all experiments was negIigible. (2) Heat loss from the respiratory tract was negligible. (3) The skin temperature wa,s constant, and equal to the ba,th temperature; therefore, the insulation value of the fur was lost. (a) The specific hea,t of the tissues was 0.83. (5) A respiratory quotient of 0.90 was assumed. (6) Surfa,ce area, (SA) calculations were made with the formula: SA : Weight,0'73. (?) Mean body temperature (MBT) was calculatedas follows: MBT:0.8 Tr*0.1 Ts (Burton, 1955). (8) Theinitial (IRH) and final body heat (X'BH) values were determined by multiplying the MBT by the specific heat of tissues and body weight. (9) Body hea,t loss (8H1,) was determined by the difference between IBII and X'RII. (10) Total heat loss (TI{L) was equal to BHL plus metabolic heat (M).
RXSULTS

The cha,nges in rectal temperature while swimming describe 25o C water as a severe cold stress for all animals studied, 30' C as a moderate cold stress for C. lateral'is and C. sp'ilosoma,33'C bath as a neutral temperature for C.lateralis and
C.spt'ilosoma

bul a moderate cold stress for M.a,uratus and D" orcli. The 36'C water bath represents a moderate heat stress for C. lateral,is and C. siti,losoma. (Figs. 1, 2,3, 4). The responses of the animals to severe cold, 25o C water, are of major interest.
The greatest heat loss was during the first' few minutes of the cold water exposure
Tabl,e 7. Mean Metabolic Rate

ol Animals in
25"C

Waterbath (ltcallhr lmz

)
animals 25" C
14.6

Animal O. lateralis
C.
spil,osoma

species

Free swimming animals

300c
r 1.8

330c
12.9

13.6

*(2.65)
10.1
(

(2.28)

(0.74)
10.8

(3.38)
TL.2

M.

1.7e)

auratus

12.2

r0.7 (2.67) r0.9

(1.23) 6.52

(2.63)

TI.2
(0.73)
9.9

(0.89)

1.40) 4.03 (0.67)

(

D. ordi

8. ?3

(1.26)

(1.24)

(0.52)

5. r0 (r.6e)

(One standard deviation)

Water Bath Studies of Mamrnals

893

(Figs. 1, 2, 3). The lateralis had least change in rectal temperature because of higher metabolic rates (Table l). The experiments cond,ucted for 2,4,6, 8 and. l0 min intervals demonstrate that maximum ra,te of heat loss for all animals occurs during
25"C
Swim:ninq

x--x H (H &-{

loterolis spilosomo ouroius

=8, n=5 =3, n=3 =2, n =2

ordi

o
F

x-x loterolis i=12,n=6 H spilosomo l=ll,n=6 H ourolus t = 3,n= 3 ""

;-;
E

;;;;

;= 3:;=

3
J0

I I

ro
Min

20

r
o

33"C

Swim

)e-x

H (H +-a

loterolis
sPilosomo

t=6,n=6
t=6,

ourotus
ord i zo Min

t=3,n=3 t=3,n=3

n =6

x-x loierolis t =3' n =3 H spilosomo I =3, n =3

X're. I Mean rectal l,emperal,ure of animals swimming in water

(f : number of swim t'rials;
25oC Confined
rz

bat'hs

- number of animals)
25"C
Deod

O a
o

F-{ loterolis I =2, n =2 H sPilosomo I =2,n=2 o---o ourotus 1"2,n=2

o a o c o o

E.

Frc. 2. Mean rectal temperature of confined living and dead animals after 10, 20 and 30 min immersion in water (l : number of trials; zz : number of animals)

894

M. L. RrnossEL.AND J. T. ynlvnnroN
Table 2. Mean Total Heat Loss ol Animals in 2so

water B0

Min (hcallhr/m2)

Species

Swimming

20.9

*(1.5) Confined
18.8

r9.2
(

19.5

15.

1.68)
6.7

(1.21)

(0.e4)

Il.4
(r.56)
9.41

12.r
(1.11)
9.1

(r.80)

(2.7t)
9.32

I1.3
(0.87)

(0.00)
9.9

(0.r0)
10.

(0.e1)
6.0 3.0

Swimming minus dead Confined minus dead

9.6 7.5

i.4

2.0

* (Standard deviation) the first 2 min of the exposure. The difference betlreen the dead and swimming a,nimals demonstrates the facilitation of heat loss by the blood circulation at least during the first 4 to 5 min (X'ig. B, Table 2).

F
E E
.E

Swimming Animo

ls

| = lolerolis = spilosomo o = ou/otu9 o = ordi

s

Fre. 3. Mean change in rectal temperature of swimming and dead animals after short term irnmersion in water (3 animals of each species at each time
int'erval)

Water Bath Studies of Mammals

895

There was no visual evidence of shivering when the animals were swimming but shivering began within I min after the animals were removed, from the water as evidenced bv visual observation and handling of the animals. The capacity for shivering was &n important factor in det'ermining the extent of heat loss of the animals confined during immersion (Figs. 5, 6). When confined, the lateralis and
First Period
Second & Third Periods

First, Second

Third Periods

l= s= o=

loterolis
ourotus
spilosomo

First

Period

Second

I Third Periods

First, Second I Third Periods

o
o E o F

t=4'n=31

ll

I I

r=roteroris
s= spilosomo o= ourotus o= ordi

E
t o o c c c
o o

t=3,

n=3'

t=2'n=2 Confined Animols

Second

c)

First

Period

I Third Periods

First, Second & Third Periods

o= ourotus o= ordi

s=

lolerolis
spilosomo

t=2.n=2 '

Deod Animols

s.D.=o.o

Fre. 4. Mean change in rectal temperatur:e of animals during three swirn

periods of l0 minutes duration in 25' C water (l : number of swim lria"ls; m : number of animals)

E96

M. L. RrnorsEr,

AND

J. T. YnlvontoN

ON

NNN

E
Y

rt

ot

NNN

m
l

Swimming

Animols

Confined Animols

Deod Animols

r
.9

Swimming Animols

Confined Animols

Deod Animols

Fre. 5. llody heat loss and meta,bolic rate during 30 min immersion in 25'C water (, : number of swim t'rials; m : number of animals) spilosoma were a,ble to maintain core temperature by a high metabolic rate but the a,uratus and ord.i were unable to preyent lowering of rectal temperature, partic-

N E E

otero I is

Y
.J o q

o o o

spilosomo ou roiu s ordi

E o F
Swimming

Animols

Confined Animols
30

Deod Animols
25o C

Tre. 6. Total heat loss of animals during

min immersion in

water

(kcal/brim'z)

Water Bath Studies of Mammals

897

ularly during the first ten-minute-swim period (X'ig. ). The metabolic rates of the confined animals confirm the visual evidence of shivering by lateralis and spilosoma (Table 1). Shivering, an important mechanism for rewarming during arousal from hibernation, is apparently weli developed in lateralis and spilosoma. Two additional points to be noted are: (1) Differences in body weight did not effect differences in the heat loss of the dead animals over a 30-min period (Fig. 6, Table 2) (2) The live animals were in or near thermal balance during the last two swim periods (Fig. a).
DISCUSSION

These water bath studies permit comparison of the capacity of animals to respond to thermal stress by shivering, increased gross activity and vasomotor activity. The curlres in Figs. I and 2 are similar and asymptotic. The bath temperature, body mass and form, and initial body temperature appear to be major factors in determining the extent of the drop in body temperature which occurs within the first 2 min. The capacity of a,nimals to increase metabolic rate under hypothermic conditions was important in determining the shape of the curve a,fter the first few

minutes. All animals had the highest activity during the first few minutes of submersion and became docile and easy to handle after 10 min in the water. There are several points of interest regarding the heat loss of animals during 30-min immersion in 25" C water. The srvimming lateralis, spilosoma, and auratus (Fig. 6) had verv similar lotalheat' loss (THL) (tt:0.2-0.7). When confined, the lateralis and spilosoma had much greater THL than auratus and ordi (Fig. 5) (p : O.Ot-0.05) primarily because of a greater metabolic rate (Ta,ble 1, Fig. 5). The auratus and ordi have limited capacity for shivering when compared to lateralis and spilosoma. Hvpothermia was undoubtedly an important faclor in limiting the metabolic rate of free swimming and confined ordi. The THL of spilosoma and lateralis were similar when confined and when swimming (Fig. 6, Table 2); however, as a result of shivering, the confined animals had less lowering of rectal temperature during the first swim period (Fig. 4). Shivering was more effective than swimming in maintaining core temper&ture, presumably beeause the metabolic rate was higher when shivering (Horvath et al., f 956). The capacity of auratus and ordi to resist lowering of body temperature when in 25" C water is limited, as evidenced by the similarity of the THL of confined and dead animals (Table 2). The second and third. periods of swimming and confined animals represent steady state conditions (X'ig. 4). During this thermal balance, total heat loss (THL) is equal to M plus BHL and is the sum of heat conducta,nce through tissues and heat transferred. by the circulation of blood from the warmer cote to the cooler periphery. The data collected on dead animals provides information regarding the rate of heat loss by tissue conductance. The thermal gradients within dead and live confined animals are undoubtedly different; however, by assuming that the conductance of heat through live and dead tissues is similar at a given rectal to water

898

M. L. RrsnnsEr, AND J. T. Yar,vsnroN

bath temperature gradient, it is possible to estimate the amount of heat transferred by tissue conductance, and to estimate the amount of heat transferred by circulation of blood. In Table 3 we have compared the rate of heat transfer of the swimming, confined, and dead animals. The rate of heat transfer per unit surface area for dead, animals of each species, given in Table 3, was the rate which occurred over the mean temTable 3. Mean Heat Loss of Amimals ,in 25" C Water during Seiond, amd, Third, Per,iod.s (kcal,lhr/rn2)

Total
heat
Loss swim.
Species and Range oC

3 Heat, loss dead

Total
heat
loss a

animals

1
b

less 3a

less

3b

confined

Swim.

Confined

C. Iateralis Swimming 33.0-28.0

Confined 37.5-32.3

I4.5

19. ?

r4.9

C, spilosoma Swimming 28.5-26.0

Confined 33.8-29.3

10.8

I 1.4

17.8

M. auratus Swimming 29.4-27.2 Confined 29.7-28.0 Swimming 28.7*26.6

Confined 30.2-27.9

t 3.8

pera,ture gradient (mean rectal less water bath temperature) existing during the steady state condition ofthe second and third swim periods. X'or instance, the rectal tempera,ture of slrimming lateralis during the second and third swim periods ranged from 28.0 to 33.0' C; therefore, the heat loss of the dead animals, 8.b kcalfhrf mz, in this temperature range is given in Table 3 and compared with the rate of THL of the swimming animals. The heat loss for all animals while swimming was greater than the heat loss of dead animals: lateralis, 40 per cent greater; spilosoma, 65 per cent; auratus, 60 per cent; and ordi,46 per cent. These differences are related to the volume of blood flow to the skin. Blood flow accounted for 6.0 and 4.2 kcallhrlm2 heat loss in lateralis and ordi while blood flow was effective in losing 7.0 and 8.2kcallfu lm2 in spilosoma and auratus. When considering the THL of confined and dead animals over the same temperature gradient, it is ofinterest to note that blood flow accounted for 20 per cent

Water Bath Studies of Mammals

899

of the hea,t loss of the confined lateralis. The greater heat loss of dead anim&ls as compared- to confined, animals of the spilosoma', auratus, and ordi species must be explained by differences in the thermal gradients of the dead and confined animals. fn a dead animal the thermal gradient is nearly equal throughout the body, whereas in a live animal peripheral tissues cool to a lower temperature than in the dead animal. With a given RI{L greater peripheral cooling means less heat is lost from the core, or in other words, there is heat loss from the periphery in preference to heat loss from the core. Estimation of the extent to which preferential cooling occurs in lateralis is not possible from the data available. That preferential peripheral cooling is verSr important, is evidenced. by heat loss of confined animals being 80 per cent, auratus, 34 per cent, spilosoma, and 33 per cent, ordi, Iess than heat loss of dead animals of the same species. trn summary, the major contribution of these wa,ter bath st'udies has beerr to point out physiological mechanisms which are operative in determining the tolerance of a given species to a given environmental stress. The C. lateral'is and C. spilosoma responded to cold water primarily by increasing metabolic rate. The spilosoma were .by cooling peripheral tissues. The M. auratus effective in limiting cooling of the core and D. ord,,ihave limited capacity for shivering but confined auratus demonst'rated conservation of body heat by peripheral cooling. It is da,ngerous at t'his time to say that a,ny given physiological response is responsible for survival of a given species in a given environment. Ilowever, capacity for shivering and peripherai cooling are important physiological mechanisms in determining the capacity of a,nimals to tolerate short term cold stress.

REFERDNCES

Alolrn,

E. F. antl R,rcrnroNo, J. (f 956) Adaptation to cold in golden hamster and ground squirrel measured chiefly by rates of body cooling. J. appl. Phgsi,ol.9' 53 - 58' Bunrow, A. C. and Eouor,rr, O. G. (1955) Man in a Cold, Enu'ironment. Monographs of the Physiological Society, No. 2. Edward Arnold, London, p. 23. IIeNNox, J. P. and VrrnECr, E. (ed.) (1955) Proceedings, S;znposia on Arctic Biology and Medicine, II. Comparative Physiology of Temperature R,egulation, Arctic Aerornedical Laborai,ory, Fort Wainwright', Alaska. Ifonvaru, s. M., srunn, G. 8., T{urr, B. K. and I{lurr,ron, L. H. (1956) Metabolic cost of shivering. J. appl. Physi,ol.8, 595 - 602. Invrwe, L. and Kaoe, J. ( 1954) Temperature of skin in the arctic as a regulator of .heat. J . appl.
Physi,ol. 7, 355 - 364. I(evsnn, C. (1961) The Physi,otogy of Natura,l Hi,bernat'ion fnternational series of monographs on pure and applied biology. Pergamon Press, London, p' 109. Pnossrn, E. L. and BnowN, F. A. (ed.) (196I\ comparati,ue Animal Phgsi.ology. w. B. Saunders, Philadelphia, London. W$,rrn, C. G. (195?) Influence of temperature on performance in guinea ptgs. Amer. J. Physi,ol,,

100,457-458.