Quarterly Journal of Experimental Physiology (1987) 72, 593-600 Printed in Great Britain

FLUID BALANCE IN FOOD-DEPRIVED LACTATING GOATS DRINKING SALINE

KRISTINA DAHLBORN
The Swedish University of Agricultural Sciences, Department of Animal Physiology, P.O. Box 7045, S-750 07 Uppsala, Sweden
(RECEIVED FOR PUBLICATION II DECEMBER 1986)

SUMMARY

Fluid balance was studied in four lactating goats during two 3 week periods, which included 30 h periods of food deprivation. In one period the goats were given 0 9 % NaCl to drink, and in the other they were given water. Prior to food deprivation, fluid intake and urinary flow were similar in the two groups, but urinary Na+ excretion was higher in the saline-drinking goats. The plasma renin activity was depressed in saline-drinking goats, while the plasma aldosterone concentration was the same both in saline-drinking and water-drinking animals. Food deprivation depressed fluid intake and urine flow in all goats, but the reduction was more pronounced in goats drinking saline. The urinary Na+ and K+ excretion also decreased, in both groups, as did plasma Na+ concentration and osmolality. The plasma protein concentration increased in both groups, indicating that hypovolaemia had developed. The renin-angiotensinaldosterone system (r.a.a.s.) became activated in goats drinking water, but not in the group drinking saline. It is suggested that sodium retention may have attenuated the activation of the r.a.a.s. in the latter group. The results of this study show that hyponatraemic hypovolaemia develops during starvation in lactating goats, regardless of the sodium state of the animals. The possibility that the hyponatraemic hypovolaemia is secondary to an impeded Na+ and fluid absorption from the rumen reticulum is discussed.
INTRODUCTION

Starvation leads to decreased water intake and negative water balance in most species, including man (Wolf, 1958), rat (Adolph, 1947), dog (Kleitman, 1927) and goat (Chaiyabutr, Faulkner & Peaker, 1980). Food deprivation also results in hypovolaemia, and Bloom & Mitchell (1960) reported from their studies in man that the blood volume reduction is accompanied by urinary Na+ losses. If humans are given entero-soluble sodium chloride tablets during fasting, blood volume reduction is attenuated, but natriuresis still occurs (Maagoe, 1968). Similarly, if rats are given saline instead of water to drink during starvation, the hypovolaemia becomes less pronounced (Wright, Reynolds, Kenny & Donlon, 1978). The reports concerning the role of the renin-angiotensin-aldosterone system (r.a.a.s.) during starvation are conflicting (Spark, Arky, Boulter, Saudek & O'Brian, 1975). Recent experiments in goats demonstrate that 26 h of food deprivation can cause hyponatraemic hypovolaemia and activation of the r.a.a.s. When the goats are given a load of water intraruminally after this period of food deprivation, the r.a.a.s. becomes further activated while a similar load of saline reduces this activity. The plasma volume and Na+ concentration increase towards normal immediately after a saline load, but not after water (Dahlborn & Karlberg, 1986). To study whether saline drinking can attenuate or prevent the development of hyponatraemic hypovolaemia during starvation in ruminants, experiments were designed in which
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the same goats were subjected to two 3 week periods, which included 30 h of food withdrawal. In one period they were given isotonic saline to drink, and in the other water. Chaiyabutr et al. (1980) have studied the cardiovascular changes which develop during starvation. They suggested that a decreased blood volume and reduced mammary blood flow could reduce milk secretion. Therefore, these stdies were done with lactating goats to investigate if saline drinking would delay the reduction of milk secretion which occurs during food deprivation.
METHODS

Animals Four lactating Swedish Landrace goats aged 1-2 years were used. The animals were kept in metabolism cages and were fed 400 g hay and 200 g grain (with 3 g of NaCl added) at 8.00 and 16.00 h, except during food deprivation. The hay contained 870 g dry matter/kg, 8-25 MJ of metabolizable energy/kg and 98 g digestible crude protein/kg, and the grain 900 g dry matter/kg, 117 MJ of metabolizable energy/kg and 140 g digestible crude protein/kg. The experiments were made 2-8 weeks after parturition and the mean milk production was 1-2 + 01 I/d. Experimental procedures The goats were investigated during 6 weeks. During the first 3 weeks two goats were given 0 9 % NaCI to drink and the other two were given water. During the second 3 week period, the goats were changed between the two treatments. Fluid intake, and urinary and milk volumes were measured twice daily at feeding times, and samples were taken for analysis. The means for the last five days before food withdrawal were taken as the control values for these parameters. During the second 3 week period faeces was collected at feeding times. Body weight was measured and a jugular blood sample taken at 10.00 h on day 15 in each 3 week period. These values were denoted as control values. On day 16 no food was given at 8.00 h (= 0 h). The goats were weighed at 8.00 h. Blood samples were taken and liquid consumption and urinary excretion measured at 2, 4, 6, 8,12, 16, 22, 24 (blood sample omitted), 26, 28 and 30 h. Food was given at 30 h and at 48 h, and the liquid consumption and urinary and milk volumes recorded (refeeding period). A final blood sample was taken at 50 h. Analysis For analysis of total plasma proteins, plasma Na+ and K+ concentration and the plasma osmolality, the blood was taken in tubes containing Li+-heparin. Total plasma proteins was measured on a TS refractometer. Na+ and K+ were analysed on an Electrolyte Analyzer (System E2A), and the osmolality on an Advanced Instrument osmometer. For determination of the plasma renin activity and the plasma aldosterone concentration blood was collected in pre-chilled tubes containing K3EDTA and then spun at +4 'C. The plasma was separated and stored at -80'C until analysed. Plasma renin activity was determined by the radioimmunoassay method described by Fyhrquist, Soveri, Puutula & Stenman (1976), and plasma aldosterone concentration by the method of McKenzie & Clements (1974). Values are presented as the mean and standard error of the mean. Test for significance was done with paired Student's t test.
RESULTS

The water-drinking group weighed 30-0 + 1'4 kg, and the saline-drinking group 31-0+ 1 6 kg before food withdrawal. The body weight decreased gradually during food deprivation, with the greater change in goats drinking saline. After 30 h the weight in waterdrinking goats had decreased to 26-6 + I-3 kg (P < 0 001) and in the saline-drinking group to 26-8 + 1l7 kg (P <0-0001). There were no significant differences in fluid intake and urine flow between groups during the control period (Fig. 1). Three main drinking occasions were seen in the water-drinking
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Control 10
0-8

Food deprivation

Refeeding

~0-6

02,,_,

1 .1 T-------A
-

A,

0-2.

FLil
/4
8 12

16

20

24

28''

Time (h)

Fig. 1. Changes in fluid intake and urine flow during 30 h of food deprivation. Dashed lines, water-drinking goats; continuous lines, saline-drinking goats (mean + s.E.M.; n = 4).

goats, namely after 0-4, 6-12 and 22-28 h. The mean water consumption during the whole food deprivation was not significantly altered as compared to the control period, but the water intake during 24-26 h was significantly less than the amount drank at 0-2 h (P < 0-01). In saline-drinking goats fluid intake was lowered during the whole of the food deprivation period compared to the control (P <0-01). After refeeding the fluid intake increased (towards controls) in saline-drinking animals, but was depressed when they drank water (P < 0-001). Urine flow had decreased after 12 h in the saline-drinking group and after 22 h of food deprivation in the water-drinking group and then remained low (P < 0 001), but if the mean urine flow was calculated for the whole deprivation period it was significantly depressed only in saline-drinking goats (P < 0-01). After refeeding, urine flow remained lower than the control period in both groups (P < 0-01). The urinary Na+ excretion was significantly greater in goats drinking saline instead of water (P < 0-05) during the control period (Fig. 2). The calculated daily Na+ intake was about 275 mmol larger than the output (urinary, faecal and milk fluid losses) in salinedrinking goats, whereas Na+ input and output were balanced in water-drinking goats. The mean urinary K+ excretion was similar in both groups (Fig. 2). The Na+ excretion was significantly depressed after 2 h and then fell gradually throughout the deprivation period in goats drinking saline (P <0-05-0-001). The fall in urinary Na+ excretion became significant after 6 h in the water-drinking group, and then remained lowered throughout the
20
EPH

72

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40 Control

K. DAHLBORN

Food deprivation

Refeeding

35
0

30 -3

25

20-

10) +~~~~~~~~

15

~ 0

'T- -71--

-_,__ -

,___ _
-

-_

15
10

215~~~~4
T
U

121

42

N;~~~~~
Time (h)

Fig. 2. Changes in urinary Na+ and K+ excretion during 30 h of food deprivation. Dashed lines, water-drinking goats; continuous lines, saline-drinking goats (mean S.E.M.; n =4).

deprivation period (P < 0-05-0 001). Urinary K+ excretion fell during food deprivation in both groups (P < 0-5-0 001). After refeeding, the urinary Na+ and K+ excretion increased to the control level in saline-drinking animals, but remained lower when they drank water
(P <OO1).
The faecal Na+ excretion decreased in the two saline-drinking goats and in one of the water-drinking goats, while the other water-drinking goat excreted the same amount of Na+ as before the deprivation period (Table 1). Milk yield decreased during starvation from S 1 + 2 to 25 + 4 ml/h in goats drinking water (P < 0001) and from 5 2 +6 to 25 +4 ml/h when they drank saline (P < 0O1). Total plasma proteins increased during starvation in both groups (Fig. 3A). After refeeding total plasma proteins returned to control levels. The plasma Na+ concentration and plasma osmolality had fallen after 2 h starvation in all animals (Fig. 3 B and C). In water-drinking goats the lowest values were obtained after 16 h. In saline-drinking goats the plasma Na+ concentration reached its lowest value after 4 h, and thereafter it increased slightly. The plasma osmolality in this group also reached its
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Table 1. Change in faecal amount and sodium excretion during 30 h offood deprivation
Water-drinking
Goat 1
Goat 2

Saline-drinking
Goat 3 35 4 41

Goat 4
28 3 37 395 12-9 43 039

Control Faeces (g/h) Dry matter (%) Na+ (mmol/h)

30 h of food deprivation Faeces (g/h) Dry matter (%) Na+ (mmol/h) A

26-6 38 047 17-2 36 007

30 4 35 0-23
16-6 39 0-23

2d14
26-6 41 097

Control .S 70
0.

Food deprivation

Refeeding

o
0

60

155

150
0

145-

Z140j
CU

300

E 295

285
50 22 262830 12 16 Time (h) Fig. 3. Changes in total plasma proteins (A), plasma Na+ (B) and plasma osmolality (C) during 30 h of food deprivation. 0, water-drinking goats; 0, saline-drinking goats (mean S.E.M.; n =4). * P < 0-05; ** P < 0-01; * P < 0 001. Values vs. the control period.
-22

2 4 6 8

20-2
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Control ,,

K. DAHLBORN

Food deprivation

Refeeding

053 0*4

023
1R0 2
x
/

700

600

,500
0

400-

0
0

200100

12 16 22 26 28 30 50 Time (h) Fig. 4. Effects of 30 h of food deprivation on plasma renin activity and plasma aldosterone concentration. 0, water-drinking goats; 0, saline-drinking goats (mean + S.E.M.; n = 4. * P < 0-05; ** P < 0-01. Values vs. the control period.

-22

2 4 6 8

lowest value after 4 h, but then increased so that after 16 h it was no longer significantly lower than during the control period. Before food withdrawal plasma renin activity was higher when the goats drank water compared to saline (P <0-01; Fig. 4). During food deprivation plasma renin activity increased after 6 h in water-drinking goats, and then remained elevated even after refeeding. One out of four goats drinking saline showed an increased plasma renin activity during food deprivation, while there was no change in the other three animals. Plasma aldosterone concentration was the same in both groups during the control period. In water-drinking animals plasma aldosterone concentration increased towards the end of the deprivation period, and it remained high also after refeeding (Fig. 4). In salinedrinking animals the plasma aldosterone concentration did not change significantly during food deprivation, although the goat showing an increased plasma renin activity also had an elevated plasma aldosterone concentration.
DISCUSSION

In the lactating goats studied here saline drinking caused neither increased fluid intake nor increased urinary flow, in contrast to observations in non-lactating sheep (Potter, 1963;
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Tomas & Potter, 1975). The goats, like the sheep (Tomas & Potter, 1975), maintained their plasma Na+ concentration and plasma osmolality. The goats in both groups had a drinking pattern related to regular feeding times also during starvation. In saline-drinking goats the mean fluid intake was depressed during the whole deprivation period, and a significant reduction of the water intake was seen in the morning after 24 h starvation in water-drinking goats. Thus, food deprivation in lactating goats depresses water intake, as has previously been reported by Chaiyabutr et al. (1980), and an even more pronounced effect was obtained in goats drinking saline. Natriuresis occurs during food deprivation in monogastric species (Bloom & Mitchell, 1960; Cizek, Simchon & Nocenti, 1977), but to the best of our knowledge, no natriuresis during starvation has been reported in ruminants. In the goats studied here the renal Na+ excretion fell gradually throughout the deprivation period and reached low values in both groups at the end of this period. There was a decreased faecal Na+ excretion and a diminished milk secretion which also reduced the Na+ losses from the body. It has earlier been shown that both the plasma volume and the glomerular filtration rate decrease during food deprivation in goats (Dahlborn & Karlberg, 1986). The increase in the total plasma protein concentration seen here indicates that the plasma volume decreased. This, in addition to the lowered urinary Na+ and K+ excretion, implies that the glomerular filtration rate fell also in the present study. Thus, several effects seem aimed at preservation of the Na+ content of the body fluids during starvation. However, the plasma Na+ concentration fell after the first meal had been omitted in both groups, and then remained low throughout the deprivation period, although plasma Na+ concentration and plasma osmolality were somewhat higher in saline-drinking than in water-drinking goats. The fact that intravascular dehydration occurs during starvation in rats drinking water, but not in rats drinking saline, led Wright et al. (1978) to draw the conclusion that rats appear to terminate drinking prematurely in order to avoid cellular overhydration when sodium is unavailable. In ruminants there is an important internal circulation of Na+ via blood plasma-salivarumen fluid-blood plasma. Dobson (1959) estimated the normal Na+ absorption from the reticulo-rumen in the sheep to be 720 mmol/d. One explanation for the development of hyponatraemic hypovolaemia in food-deprived goats may be a slower absorption of Na+ and fluid from the rumen to plasma during starvation. This has been reported in sheep (Stacy & Warner, 1966), and in goats K. Holtenius & K. Dahlborn (submitted for publication) have shown that the rumen fluid Na+ concentration increases and the net absorption of fluid from the rumen becomes depressed. Because the hyponatraemia, and probably also the hypovolaemia, were of a similar magnitude in both groups, neither the fall in plasma Na+ concentration nor the hypovolaemia seemed to be the primary cause of the activation of the r.a.a.s. which was seen in waterdrinking goats (Dahlborn & Karlberg, 1986). Here, the r.a.a.s. was activated in one of the four saline-drinking goats, but not in the other three, which indicates that some factor other than hyponatraemia and hypovolaemia must have been responsible. A possible explanation could be that in saline-drinking goats the accumulated Na+ content of the body inhibited activation of the r.a.a.s. as it has been shown that the total Na+ content of the body rather than the plasma Na+ concentration determines the activity of the r.a.a.s. (Denton, 1982). In addition the lack of activation of the r.a.a.s. in starving saline-drinking goats could be due to a greater load of NaCl reaching the macula densa cells of the kidneys in these animals (Ganong, 1985). From the results obtained in this study, it seems justified to assume that the blood flow to
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the mammary gland decreased to a similar extent in both groups. Therefore the observation that the reduction of the milk secretion was similar was not unexpected. In conclusion, starvation was accompanied by hypovolaemia in ruminants, as previously shown for monogastric species. However, in ruminants this effect was not secondary to increased Na+ losses via the urine and could not be prevented by saline consumption, but rather appeared to be caused by an impaired Na+ and fluid absorption from the rumen.
This study was supported by the Swedish Council for Forestry and Agricultural Research (Project 612/82).
REFERENCES ADOLPH, E. F. (1947). Urges to eatand drink in rats. The American Journal of Physiology 151, 110-125. BLOOM, W. L. & MITCHELL, W. (1960). Salt excretion of fasting patients. Archives of Internal Medicine 106, 321-326. CHAIYABUTR, N., FAULKNER, A. & PEAKER, M. (1980). Effects of starvation on the cardiovascular system, water balance and milk secretion in lactating goats. Research in Veterinary Science 28, 291-295. CIZEK, L. J., SIMCHON, S. & NOCENTI, M. R. (1977). Effects of fasting on plasma volume and fluid and sodium exchanges in male rabbits. Proceedings of the Society for Experimental Biology and Medicine 154, 299-303. DAHLBORN, K. & Karlberg, B. E. (1986). Fluid balance during food deprivation and after intraruminal loads of water or isotonic saline in lactating and anoestral goats. Quarterly Journal of Experimental Physiology 71, 223-233. DENTON, D. (1982). The Hunger for Salt. An Anthropological, Physiological and Medical Analysis. Berlin, Heidelberg, New York: Springer-Verlag. DOBSON, A. (1959). Active transport through the epithelium of the reticulo-rumen sac. Journal of Physiology 146, 235-251. FYHRQUIST, F., SOVERI, P., PUUTULA, L. & STENMAN, U. H. (1976). Radioimmunoassay of plasma renin activity. Clinical Chemistry 22, 250-256. GANONG, W. F. (1985). Review of Medical Physiology. ed. DRAWER, L., Los Altos, CA, U.S.A.: Lange Medical Publications. KLEITMAN, N. (1972). The effect of starvation on the daily consumption of water by the dog. American Journal of Physiology 81, 336-340. MAAGOE, H. (1968). Changes in blood volume during absolute fasting with and without sodium chloride administration. Metabolism 17, 133-138. MCKENZIE, J. K. & CLEMENTS, J. A. (1974). Simplified radioimmunoassay for serum aldosterone utilizing increased antibody specificity. Journal of Endocrinology and Metabolism 38, 622-627. POTTER, B. J. (1963). The effect of saline water on kidney tublular function and electrolyte excretion in sheep. Australian Journal of Agricultural Research 14, 518-528. SPARK, R. F., ARKY, R. A., BOULTER, P. R., SAUDEK, C. D. & O'BRIAN, J. T. (1975). Renin, aldosterone and glucagon in the natriuresis of fasting. Seminars in medicine of the Beth Israel Hospital, Boston. New England Journal of Medicine 292, 1333-1340. STACY, B. D. & WARNER, A. C. I. (1966). Balances of water and sodium in the rumen during feeding: osmotic stimulation of sodium absorption in the sheep. Quarterly Journal of Experimental Physiology 51, 79-93. TOMAS, F. M. & POTTER, B. J. (1975). Influence of saline drinking water on the flow and mineral composition of saliva and rumen fluid of sheep. Australian Journal of Agricultural Research 26, 585-598. WOLF, A. V. (1958). Thirst. Physiology of the Urge to Drink and Problems of Water Lack. Springfield, IL, U.S.A.: Charles C. Thomas Publisher. WRIGHT, J. W., REYNOLDS, T. J., KENNY, J. T. & DONLON, K. (1978). Inhibition of food deprivation induced hypovolemi.- by saline consumption in rats. Physiology and Behavior 21, 215-221.

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