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Jan Marczuk, Lukasz Kurek Krzysztof Lutnicki ©
Department of Internal Medicine of Farm Animals and Horses Chair and Clinic of Internal Medicine University of Life Sciences in Lublin ul. Glqboka 30 20 - 612 Lublin, Poland e-mail: doktorjm@o2.pl
BIOCHEMICAL CHANGES IN BLOOD IN THE COURSE OF SUBCLINICAL AND CLINICAL FORMS OF FATTY LIVER SYNDROME
IN DAIRY COWS
Summary
Fatty liver syndrome in dairy cows is characterized by excess triglyceride (TG) accumulation in the liver and apparent decreased hepatic lipoprotein output. The aim of this work was to assess carbohydrate-lipid metabolism parameters and to investigate the liver function status indicators. Investigations were carried out on 36 dairy cows, black-white breed, which were in the last month of pregnancy period. The animals were divided into two groups- the experimental one consisting of 24 animals with fatness in drying period, and 12 cows at normal state of nutrition as a control group. In both groups clinical examinations and biochemical estimations were carried out in 4th and 2nd weeks to labours as well as in 1-2, 4-5 and 7-8 weeks after parturition respectively. In blood the concentration of ketone bodies (KB), free fatty acids (FFA), glucose, total cholesterol and triglicerids, total bilirubin and enzyme AST, GGT, GLDH activity was estimated. In extremely fatness it comes in cows to subclinical form of fatty liver syndrome after parturition. The clinical form of this disease was always fatal and pathological examination showed varied degree of liver steatosis. The GLDH, AST activity and total bilirubin were increased in sublinical and clinical forms of above mentioned syndrome.
Additionally, there were 12 cows included in the study treated in our clinic due to excessive lipid mobilization in the course of fatty liver syndrome, being in different periods after parturition - as a another experimental group (clinical trial).
Key words: fatty liver syndrome, dairy cows, parturition, carbohydrate and lipid disorders
Introduction.
Metabolic diseases in dairy cows occur predominantly in peripartum period and are suggested to be derived from fatty liver initially developed in drying time ( ). Hepatic uptake of free fatty acids (FFA) released in excess from adipose tissues in negative energy balance conditions and impaired lipoprotein metabolism in the liver is crucial for developing of this illness.
The fatal influence of extreme fatness in drying period on the health of dairy cows after labour has been well recognized from about 30 years. It may cause fatal
© Jan Marczuk, Lukasz Kurek Krzysztof Lutnicki, 2008
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disease named fatty liver syndrome, fat cow syndrome or extreme fatty mobilization syndrome, as well steatosis hepatis. The clinical form of this disease exist predominantly only in single cows, whereas subclinical form usually concern 50-90% of the herd, being the most particular economic problem in animal husbandry due to ineffective treatment, which lead to cow elimination without the possibility to its effective exploitation according to their genetic potency to milk production. Subclinical form of this syndrome leads to metabolic disorders and infections susceptibility, rise in milk production, conditional worsening and persisting fertility disorders.
Mainly subclinical character of this disease and the fact that the clinical form has no characteristic nor specific signs make this disease difficult to recognize, so the diagnosis is predominantly (generally) based on laboratory analysis and liver biopsy.
Bibliographical data concerning this disease show the great amounts of divergences in use of blood biochemical parameters in treatment, early diagnosis and possibility of liver function and damage estimation - especially in sublinical form of fatty liver syndrome. The aim of this work was to assess:
- energy metabolism pathways parameters in country conditions and the liver functional status in dairy cows with subclinical form of fatty liver syndrome
- analysis and comparison of the data in clinical and subclinical forms of the disease
- optimization of biochemical parameters useful to diagnose the liver damage and functional disorders in cows during postpartum period.
Material and methods.
The examinations were carried out during two winter-spring seasons in dairy cows farm herd , which consisted of 36 animals, 4-10 years old, breed black white with addition of Holstein Friesian breed blood. All animals were in the last month of pregnancy.
The first experimental group consist of 24 cows, which showed fatness during the drying period. The second experimental group consisted of 12 cows, coming from many other farms, which were treated in our clinic due to clinical or subclinical forms of liver steatosis, diagnosed post mortem. The control group consisted of 12 cows at normal state of nutrition. The average milk efficiency estimated during 305 day lactation period was 5 300 to 8 100 kg of milk. The examinations were carried out at 4 and 2 weeks to labour time and 1 -2, 4-5, 7-8 weeks after labour. Lipid-carbohydrate metabolism parameters as well as liver function status were assessed. In blood were estimated: ketone bodies(KB) concentration according to Goschke method, free fatty acids (FFA) concentration according to Dole method, glucose concentration using Alfa Diagnostic kit, triglicerids (HTL). Enzymes activity -aspartate transaminase (AST), gamma glutamate transferase GGT, glutamate dehydrogenase GLDH, were determined spectrophotometrically using manufacture kits Alfa Diagnostics and Cormay and Epoll 200 spectrophotometer according to original procedure. Liver function was determined on the basis of total bilirubine concentration according to Jendrassik and Cleghorn method, albumine concentration with help of Cormay factory kit, total cholesterol with Alpha Diagnostic kits. The
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livers obtained from dead cows affected with fatty liver syndrome were examined histopatologically with the use HE and Sudan IV staining. The data were expressed as mean (SD). Statistical analysis for significant differences was performed according to the Student's t-test for unpaired data and p value < 0,01 was considered statistically significant.
Results
The analysis of the above mentioned parameters shows, that in the groups of cows affected with fatness status during drying period (gr 1) leads to liver function injury and energy metabolism pathway disruption , markedly expressed in after labour (post partum) period .They were most particularly exhibited 1-2 weeks after labour . The similar changes were observed in patients with clinical form of fatty liver syndrome (liver steatosis), but more extensive especially in carbohydrate-lipid metabolism and liver injury (Tab.1, 2)
Table 1.
Means values of investigated parameters in blood obtained from experimental __group (farm trial) D and control group._
Parameters Cows PERIOD IN WEEKS
Before parturition After parturition
4 . 2 . 1-2 . 4 -5 . 7-8 .
Glucose [mmol/L] D 3,95 3,98 2,84**b 3,33 **a 3,38
control 4,01 3,95 3,75 3,66 3,82
FFA [mmol/L] D 0,21 0,26 1,03 **b 0,70 **b 0,40 **
control 0,22 0,23 0,39 ** 0,49 ** 0,34 **
Ketone bodies [mmol/L] D 0,47 0,41 1,77 **b 0,96 **b 0,62 **b
control 0,64 0,6 0,62 0,63 0,45
Cholesterol [mmol/L] D 3,31 3,16 2,83 **a 4,26 **b 5,36 **
control 3,26 3,32 3,59 5,84 ** 6,00 **
Triglicerides [mmol/L] D 0,29 0,27 0,08 **b 0,17 **b 0,20 **b
control 0,25 0,27 0,20 0,26 0,29
AST [U/L] D 56,6 58,4 112,2 **b 83,3 ** b 67,1 ** b
control 54,4 56,5 67,7 ** 67,3 ** 63,2 **
GGT [U/L] D 19,7 18,8 a 25,0 ** b 28,2 ** b 25,7 ** b
control 19,7 18,8 20,8 21,5 20,9
GLDH [U/L] D 5,1 5,2 18,5 ** b 15,9 ** b 8,6 ** b
control 4,9 4,9 6,5 * 6,3 * 6,5 *
Total bilirubin [mmol/L] D 3,9 4,5 14,2 ** b 10,6 ** b 5,5 ** a
control 3,6 4,1 5,1 * 4,8 4,5
Albumin [g/L] D 38,7 37,1 33,8 * b 33,6 * b 33,4 * b
control 38,0 37,7 37,0 36,2 36,9
* (p<0,05), ** (p<0,01) - statistically significant difference in comparison to cows being 2-4 weeks before parturition .
a (p<0,05), b (p<0,01) - statistically significant difference in comparison to control group.
Obtained results show, that dairy cow fatness evokeed markedly expressed negative energy balance in prelabour period and 1-2 weeks after parturition, which was characterized by increased blood FFA and KB concentration and decreased
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glucose concentration. In the group affected with subclinical form of fatty liver syndrome appeared to retrained fetal membranes (8 cows) metritis (10 cows), sublinical ketosis (18 cows), paralysis (5 cows), coexisting with liver function injury, which was expressed as increased enzymes activity (AST, GGT,GLDH) and total bilirubine concentration whereas the concentration of cholesterol, albumin and triglicerids were decreased. Cows with clinical form of fatty liver syndrome (clinical trial) exhibited the same changes in blood, but they were more exacerbated. Histopathological examination showed high degree liver steatosis. The intensity of these pathological changes and lack of successful treatment caused fatal prognosis.
Table 2.
Mean values of parameters obtained from cows treated in clinic (clinical trial), _investigated in farm herd (farm trial) and control group._
Examined parameters clinical trial farm trial control
Glucose [mmol/L] 1,98 ** 2,84 ** 3,75
FFA [mmol/L] 1,60 ** 1,03 ** 0,39
Ketone bodies [mmol/L] 3,18 ** 1 77 ** 0,62
Cholesterol [mmol/L] 2,4* 2,83 * 3,6
Triglycerides [mmol/L] 0,07 ** 0,08 ** 0,20
AST [U/L] 161,1** 112,2 ** 67,7
GGT [U/L] 43,2* 25,0 ** 20,8
GLDH [U/L] 30,4** 18,5 ** 6,5
Total bilirubin [mmol/L] 20,7** 14,2 ** 5,1
Albumine [g/L] 33,2* 33,8 * 37,0
(p<0,05), ** (p<0,01) - significant in comparison to control.
Conclusions.
1. Cows with fatness exhibit energy metabolism pathway disruption and liver function injury predominantly 1-2 weeks after labour, which indicates the sublinical course of fatty liver syndrome. In clinical form of this diseases the above mentioned data are more expressed
2. The estimations of AST,GLDH, GGT activity and total bilirubine concentrations in correlation to FFA, KB and triglycerides amount in blood of dairy cows during peripartum period are useful for early diagnoses and give a beneficial effects in treatment.
3. The above mentioned parameters were reliable indicators of the liver function and were correlated with degree of hepatic lipidosis.
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