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Clinical signs of fatty liver and kidney syndrome in broilers and their alleviation by the short‐term use of biotin or animal tallow D. Balnave

a c

b

, M. N. Berry & R. B. Cumming

a

a

Department of Biochemistry and Nutrition , University of New England , Armidale, New South Wales, 2351, Australia b

Department of Biochemistry, School of Medicine , The Flinders University of South Australia , South Australia, 5042, Australia c

New South Wales Department of Agriculture , Poultry Research Station , Seven Hills, Sydney, New South Wales, 2147, Australia Published online: 08 Nov 2007.

To cite this article: D. Balnave , M. N. Berry & R. B. Cumming (1977) Clinical signs of fatty liver and kidney syndrome in broilers and their alleviation by the short‐term use of biotin or animal tallow, British Poultry Science, 18:6, 749-753, DOI: 10.1080/00071667708416430 To link to this article: http://dx.doi.org/10.1080/00071667708416430

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Br. Poult. Sci., 18: 749-753. 1977

Longman: printed in Great Britain

CLINICAL SIGNS OF FATTY LIVER AND KIDNEY SYNDROME IN BROILERS AND THEIR ALLEVIATION BY THE SHORT-TERM USE OF BIOTIN OR ANIMAL TALLOW D. BALNAVE1, 3, M. N. BERRY2 AND R. B. CUMMING 1

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1

Department of Biochemistry and Nutrition, University of New England, Armidale, New South Wales 2351, Australia and 2 Department of Biochemistry, School of Medicine, The Flinders University of South Australia, South Australia 5042, Australia Received for publication 4th February 1977

1. Birds affected by fatty liver and kidney syndrome (FLKS) had elevated concentrations of serum Na + , K+, lactate, pyruvate and uric acid and reduced concentrations of serum HCO - 3 and glucose. 2. Short-term treatment with biotin or animal tallow reduced the mortality from FLKS and prevented the clinical signs. 3. Lactic acidosis may be a major factor contributing to the mortality and physical symptoms observed in birds affected by FLKS. The lactic acidosis and the hypoglycaemia observed in FLKS are due primarily to an accumulation of pyruvate as a result of an insufficiency of biotin for normal pyruvate carboxylase activity. INTRODUCTION

The fatty liver and kidney syndrome (FLKS), which can cause high mortality in growing broilers, is a biotin-responsive condition (Payne et al., 1974; Whitehead and Blair, 1974). It can also be prevented by supplementing FLKS-inducing diets with animal tallow for a short period of 54 h (Balnave et al., 1976; 1977a). In this latter work the changes in liver composition and enzyme activities which developed with the onset of FLKS were prevented by short-term feeding with animal tallow. Other studies of glucose metabolism indicated that birds suffering from the syndrome have significantly lower glucose pool sizes and synthesis rates and plasma glucose concentrations than birds fed on the same diet supplemented with biotin or tallow (Balnave et al., 19773). Studies have shown that FLKS induces changes in the composition of blood as well as liver (Bannister et al., 1975; Evans et al., 1975; Johnson et al., 1976; Whitehead et al., 1976). The present studies were carried out to investigate the clinical changes in blood composition induced by FLKS and to examine the role of animal tallow in preventing the syndrome. 3 Present address: New South Wales Department of Agriculture, Poultry Research Station, Seven Hills, Sydney, New South Wales 2147, Australia. 749

750

D. BALNAVE, M. N. BERRY AND R. B. CUMMING

MATERIALS AND METHODS

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Two experiments were conducted using broiler pullets from a young breeder flock which were obtained at 1 day of age from a commercial hatchery. They were fed on crushed wheat for 4 d and then on the diet shown in Table 1. Food and water were freely available. Three treatments were applied: birds maintained to day 28 on the basal diet; birds given animal tallow as a dietary supplement (150 g/kg) for the 54 h prior to starving on day 28 and birds given biotin. In experiment 1 the biotin was given as a dietary supplement of 0*3 mg/kg of diet from day 21 while in experiment 2 a total of 300 fig biotin was injected in three injections during the 54 h prior to starving on day 28. TABLE 1 Composition of basal diet (g/kg)

Crushed wheat Meat and bone meal DL-methionine L-lysine Premix1

794 200 1 -4 2-5 2-1

1 Provided the following micronutrients per kg of diet; retinol, 1-8 mg; cholecalciferol, 62-5 fig; DL-atocopherol, 8 mg; menaphthone, 2 mg; nicotinic acid, 25 mg; pantothenic acid, 10 mg; folic acid, 1-5 mg; riboflavin, 5 mg; vitamin B12, 7 fig; Mn, 80 mg.

In each experiment food, but not water, was removed for 18 h on day 28 in order to initiate the syndrome. At the end of the period of starvation blood samples were taken from surviving birds selected at random and the sera separated and used for subsequent analyses. Birds on treatment 1 were grouped depending on whether or not they showed physical signs of FLKS (classes 3 to 5, Whitehead, 1975). Blood samples were taken from each group. Lactate was measured by the method of Hohorst (1965) and pyruvate according to Holzer and Holldorf (1956). Glucose, uric acid, sodium, potassium and bicarbonate were measured by standard techniques on a Technicon multichannel autoanalyser. RESULTS AND DISCUSSION

In experiment 1 seven birds from the 35 on treatment 1 died as a result of the 18-h period of starvation on day 28 while in experiment 2 eight from the 49 died. Only one other bird (experiment 2, treatment 2) died as a result of the withdrawal of food. The results are given in Table 2. These indicate that the onset of FLKS was associated with significant increases in serum Na+, K+, lactate, pyruvate and uric acid and a significant reduction in serum glucose. Serum alanine aminotransferase was also significantly increased in birds affected with FLKS. FLKS is known to be a biotin-responsive condition and birds affected by the syndrome have low concentrations of liver biotin (Johnson et al., 1976). The elevations in serum lactate and pyruvate in birds affected by FLKS are therefore not

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TABLE 2

Serum composition (fimol/ml) in birds starved for 18 h (5 birds/treatment)

Treatment FLKS diet

Experiment

Birds showing signs of FLKS Birds without FLKS

{a

{I

FLKS diet + tallow FLKS diet+biotin bhM

{5 {I

Na+

K+

Glucose

Uric acid

162-6*** 162-6**

11-3*** 13-9***

44*** 8-4

V-45***

1490 151-4

8-3 11-5*

13-0 11-2

0-45

148-4 148-6

5.4** 10-4

12-9 10-6

0-34

145-2 149-4

8-0 8-5

130 9-6

0-20

1-77 3-07

0-51 0-78

0-69 0-93

HCO3

Pyruvate

6-0***

18-9*** 15-6***

0-34*** 0-40***

7-2***

12-8* 12-2***

009 0-22**

8-2

7-6 15-5***

0-23** 0-21*

8-4

005 004

5.2

0-034 0043

1-55 ...

12-7 13-4

0-107

Lactate

Alanine aminotransferase

1-08 ...

7-9 6-9 1-29 0-77

* P d

5 z

i

D O W

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752

D. BALNAVE, M. N. BERRY AND R. B. CUMMING

unexpected and have been reported previously (Bannister et al., 1975; Johnson et al., 1976). However, the high concentrations of Na + and K+ now reported in birds affected by FLKS, in combination with high lactate concentrations, suggest that the chemical pathology of the condition is a developing lactic acidosis which is secondary to the accumulation of pyruvate because of a biotin insufficiency. The relatively high concentration of lactate and low concentration of HCO~ in sera from birds on treatment 1 but not showing signs of FLKS is an indication that the same changes were occurring in these birds, but that the acidosis was not as pronounced as in birds affected by the syndrome. In experiment 1 both biotin and tallow prevented the lactic acidosis. However, in experiment 2 the tallow had no effect on the lactate concentrations and in both experiments the pyruvate concentrations remained partly elevated. Balnave et al. (1976; 1977a) and Johnson et al. (1976) observed that hepatic lipogenic enzyme activity is increased in birds affected by FLKS. Balnave et al. (1977a) suggested that the beneficial effect of tallow is due to its inhibiting effect on lipogenic enzyme activity with the resulting release of biotin from acetyl CoA carboxylase and availability for pyruvate carboxylase. However, the half-life of acetyl CoA carboxylase is 48 h (Majerus and Kilburn, 1969) so that it would be unlikely that a complete recovery would be observed in all variables within 54 h of dietary supplementation with tallow. Nevertheless, in this time positive responses were obtained in a reduction in mortality and in all the serum metabolites measured except serum lactate in experiment 2. The results of the present experiments are compatible with the hypothesis of Balnave et al. (1977a) that in the absence of an adequate supply of biotin the distribution of the available biotin is the major factor involved in preventing FLKS. An increased availability of biotin for pyruvate carboxylase, resulting in an increased rate of gluconeogenesis, would suffice to alleviate the clinical signs observed in the present work. Although gluconeogenesis and blood glucose concentrations are reduced in birds affected by FLKS and mortality may arise because of a lack of glucose for essential functions when birds are exposed to a stress such as fasting (Evans and Bannister, 1974; Bannister et al., 1975; Balnave et al, 19773) the present investigations suggest that mortality may also be related to a developing lactic acidosis. ACKNOWLEDGEMENTS

The authors acknowledge the skilled technical assistance of Mr J . Wolfenden and Miss Leigh Fricker. The present work was carried out when the senior author was a Visiting Research Fellow at the University of New England. Financial assistance was generously supplied by the Australian Chicken Meat Research Committee and the Council of Egg Marketing Authorities of Australia. REFERENCES BALNAVE, D., CUMMING, R. B. AND SUTHERLAND, T. M. (1976). The role of dietary fat in alleviating

fatty liver and kidney syndrome in broilers. Aust. vet. J., 52: 433-434.

FATTY LIVER AND KIDNEY SYNDROME

753

BALNAVE, D., CUMMING, R. B. AND SUTHERLAND, T. M. (1977a). A biochemical explanation for

the fatty liver and kidney syndrome of broilers; its alleviation by the short-term use of dietary fat. Br. J. Nutr. (in press). BALNAVE, D., WOLFENDEN, J., BALL, F. M., CUMMING, R. B. AND LENG, R. A. (1977b).

Studies

of fatty liver and kidney syndrome in chickens: dynamics of glucose metabolism. Br. J. Nutr. (in press).

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BANNISTER, D. W., EVANS, A. J . AND WHITEHEAD, C. C. (1975). Evidence for a lesion in carbo-

hydrate metabolism in fatty liver and kidney syndrome in chicks. Res. vet. Sci., 18: 149-156. EVANS, A. J . AND BANNISTER, D. W. (1974). Fatty liver and kidney syndrome in broilers: abnormalities in lipid and carbohydrate metabolism. Wld's Poult. Sci. J., 30: 231. EVANS, A. J., BANNISTER, D. W. AND WHITEHEAD, C. C. (1975). Some aspects of lipid metabolism in fatty liver and kidney syndrome in chicks. Res. vet. Sci., 18: 26-31. HOHORST, H. J . (1965). I n : Methods of Enzymatic Analysis, pp. 266-270. Edit. BERGMEYER, H. U . New York, Academic Press. HOLZER, A. AND HOLLDORF, A. (1957). Isolierung von D-glycerat-dehydrogenase, einige eigenschaften des enzyms und seine verwendung zur enzymatischoptischen bestimmung von hydroxypyruvat neben pyruvat. Biochem. Z., 329: 292-312. JOHNSON, A. R., HOOD, R. L., PEARSON, J . A. AND FOGERTY, A. C. (1976).

The role of biotin in

the stress-induced death of chickens exhibiting fatty liver and kidney syndrome. Proc. Nutr. Soc., 35: 129A-130A. MAJERUS, P. W. AND KILBURN, E. (1969). Acetyl CoA carboxylase. The roles of synthesis and degradation in regulation of enzyme levels in rat liver. J. biol. Chem., 244: 6254-6262. PAYNE, C. G., GILCHRIST, P., PEARSON, J . A. AND HEMSLEY, L. A. (1974).

Involvement of biotin

in the fatty liver and kidney syndrome of broilers. Br. Poult. Sci., 15: 489-498. WHITEHEAD, C. C. (1975). Tissue lipid composition in fatty liver and kidney syndrome in chicks. Res. vet. Sci., 18: 32-35. WHITEHEAD, C. C., BANNISTER, D. W., EVANS, A. J., SILLER, W. G. AND WIGHT, P. A. L. (1976).

Biotin deficiency and fatty liver and kidney syndrome in chicks given purified diets containing different fat and protein levels. Br. J. Nutr., 35: 115-125. WHITEHEAD, C. C. AND BLAIR, R. B. (1974). The involvement of biotin in the fatty liver and kidney syndrome in broiler chickens. Wld's Poult. Sci. J., 30: 231.

Clinical signs of fatty liver and kidney syndrome in broilers and their alleviation by the short-term use of biotin or animal tallow.

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