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Advances in Environmental Biology, 5(7): 1794-1798, 2011 ISSN 1995-0756 1794 This is a refereed journal and all articles are professionally screened and reviewed ORIGINAL ARTICLE Study of Fatty Liver Syndrome Frequency in Dairy Cattle by Evaluating BHBA, Urea, Total Protein, Cholesterol and Serum Values in Tabriz 1 Amirparviz Rezaeisaber, 2 Mehrdad Nazeri 1 Department of Clinical Science, Tabriz Branch, Islamic Azad University, Tabriz, Iran. 2 Young Researchers Club, Tabriz Branch, Islamic Azad University, Tabriz, Iran Amirparviz Rezaeisaber, Mehrdad Nazeri; Study of Fatty Liver Syndrome Frequency in Dairy Cattle by Evaluating BHBA, Urea, Total Protein, Cholesterol and Serum Values in Tabriz ABSTRACT Fatty liver syndrome (Hepatic lipidosis) or fat cow syndrome is a major metabolic disorder in many dairy cattle's in early period of lactation. The aim of this study was to evaluating fatty liver syndrome in dairy cattle in Tabriz by measurement of BHBA, Urea, Total Protein, cholesterol and serum values. The results showed that has a positive relationship with urea and BHBA serum values and reverse relationship with Cholesterol and Total Protein. Thus, with elevating of serum values, Urea and BHBA also increased and Cholesterol and Total Protein contrary diminished. Key words: fatty liver, dairy cattle,, BHBA, Urea, Total Protein, cholesterol. Introduction Fatty liver syndrome (Hepatic lipidosis) or fat cow syndrome is a major metabolic disorder in many dairy cattle's in early period of lactation [3,6] and it's combined with decrease in health and reproduction rate of livestock [9,14]. Fatty liver syndrome was documented in forties (decade 1940) but there were few researches about it until mid-seventies. In early 70 and 80 decades, this syndrome was reported around parturition widely and it was recorded in many countries [3,6]. When this disorder is severe, milk production and appetite of cow, both are decreased. So effective prevention of fatty liver can save millions of dollars every year and prevent from decrease in milk production [4]. Incidence of Fatty liver in dairy cattle is mainly in first four weeks after parturition [10], when more than 50% of cows show different degrees of Triacylglycerol (TAG) accumulation in their livers [13,14]. One of the reasons is that daily nutrition of cow is not sufficient and it can't meet increasing need of energy in cattle that is producing milk. In this condition, none Esterified fatty acid () is released from adipose tissue, often more than it's needed, and extra amount is transferred to liver, especially in fat cows [15]. Fatty liver occurs when liver harvesting of lipids is more than their Oxidation and secretion by the liver and it is with high plasma concentrations of that is resulted from high adipose tissue [4,10]. Extra fat is stored in liver as TAG and results in decrease of metabolic function of liver [4]. Liver is classified to three types, according to fat level: normal liver, liver with average fat and liver with very high fat [4,10]. The latter type is categorized to Nonencephalopathic fatty liver [1] and hepatic encephalopathy [4,10]. Unbalanced or insufficient nutrition, overweight and high concentration of estrogen are involved in etiology of fatty liver [9]. The disorder can be accompanied with high rate of dystocia, infectious and inflammatory disease, long interval between parturitions and reduction of milk and longevity average [9]. Forasmuch as even slight fatty liver is dual with decrease in health and reproduction status of cow, prevention of its occurrence with supplying enough food and creating an isolated place at preparation period for parturition can reduce decline rate of producing milk and it Corresponding Author: Amirparviz Rezaeisaber, Department of clinical science, Tabriz branch, Islamic Azad University, Tabriz, Iran. E-mail: aprs_1352@yahoo.com Am_rezaei@iaut.ac.ir Tel: +98 914 411 7297

would be the most efficient therapeutic procedure among the other methods [18]. However this prevention is not enough for fat cows or the ones that are not feed well, the cows that have problem during parturition or had twins, the cows that have metabolic or infectious disease and the ones that have developed severe energy imbalance because of producing high amount of milk immediately after parturition [18]. Assuming existence of about 9 million dairy cattle all over the America, annual charges of fatty liver in this country is estimated more than 60 million dollars [1]. If there are more studies about molecular changes and relationship between the disease and immunity function, better remedies and more efficient ways to prevent fatty liver can be presented [19]. In our country, because of industrial methods that speed for nurture and maintenance of dairy cattle, and because of producing more milk, more nutrition is considered; occurrence of this syndrome is most likely. According to these conditions, providing exact diagnose of this syndrome and estimate it's incidence rate and finally how to prevent it in our country is a necessity and this case made us do the first study about this disease in Tabriz. It's possible that origin of many diseases happening around parturition could be fatty liver incidence in this region's dairy cattle. Materials and Methods This research is descriptive analytical. In this quest, during frequently visits from dairy cattle farms of Tabriz, according to statistics of dairy cattle in Tabriz area, the inspection of 400 Holstein cows were done. In this inspection, age, body condition score and Pregnancy status of animals was investigated [7]. In next stage, according achieved results; Cows based on Table 1 were divided into four groups. Simultaneous inspection of animals, attempting to obtain blood samples of 10 ml of jugular vein was done by venoject. Blood samples taken near the ice and sent to the laboratory and after serum preparation were freezing inside the micro tube. At the time of testing, sera were defrosted and levels in serum by Randox kit and Auto analyzer were measured. In this study, levels of Urea, Total Protein and cholesterol in serum by Pars test kits and by spectrophotometric method was measured. In this study to analyzing and comparison of data were used of ANOVA test and to evaluate the relationship between the together, correlation test was used. Results: Serum Value Average: Based on table 2 and ANOVA test results, serum value average in group +8 was 825.60±105.23, in group +1 was 628.40±65.9, in 1795 group -8 was 728.64±35.66 and in group -1 was 1125.58±123.60 that has significant difference (P<0.001) with normal value (86.18). Urea Serum Value Average: Based on table 3 and ANOVA test results, Urea serum value average in group +8 was 16.10±2.18, in group +1 was 32.60±5.79, in group -8 was 21.52±5.07 and in group -1 was 13.00±2.86 that has significant difference (P<0.001) with normal value (49.34). BHBA Serum Value Average: Based on table 4 and ANOVA test results, BHBA serum value average in group +8 was 9.22±0.55, in group +1 was 6.41±1.34, in group -8 was 7.79±0.70 and in group -1 was 9.31±1.70 that has significant difference (P<0.001) with normal value (18.20). Cholesterol Serum Value Average: Based on table 5 and ANOVA test results, Cholesterol serum value average in group +8 was 121.60±23.61, in group +1 was 228.66±13.93, in group -8 was 158.76±29.79 and in group -1 was 84.91±11.07 that has significant difference (P<0.001) with normal value (108.58). Total Protein Serum Value Average: Based on table 5 and ANOVA test results, Total Protein serum value average in group +8 was 8.66±0.36, in group +1 was 10.24±0.87, in group -8 was 9.18±0.43 and in group -1 was 7.70±1.04 that has significant difference (P<0.001) with normal value (27.73). Relationship Between and Urea: Based on table 6 and pearson's Correlation index revealed that between and urea serum values there is a significant and direct correlation so that correlation index was r=0.71. This index indicates a positive effect of on urea serum values. Thus, with elevating of serum values the urea values also increase. Relationship Between and BHBA: Based on table 7 and pearson's Correlation index revealed that between and BHBA serum values there is a significant and direct correlation so that correlation index was r=0.61. This index indicates a positive effect of on BHBA serum values. Thus, with elevating of serum values the BHBA values also increase.

1796 Table 1: Classification of cattle based on uterine position Non-pregnant cows New born to under one month postpartum (-1) More than one month of their last calving (+1) pregnant cows Less than 8 month (-8) More than 8 month (+8) Table 2: Mean serum in Holstein dairy cows based on pregnancy status. +8 825.60±105.23 601413.04 6978.49 86.14 0.001 +1 628.40±65.9-8 728.64±35.66-1 1125.58±123.60 total 806.96±201.55 Table 3: Mean serum Urea in Holstein dairy cows based on pregnancy status. +8 16.10±2.18 1001.51 20.29 49.34 0.000 +1 32.60±5.79-8 21.52±5.07-1 13.00±2.86 total 21.70±8.71 Table 4: Mean serum BHBA in Holstein dairy cows based on pregnancy status. +8 9.22±0.55 24.66 1.35 18.20 0.000 +1 6.41±1.34-8 7.79±0.70-1 9.31±1.70 total 8.01±1.63 Table 5: Mean serum Cholesterol in Holstein dairy cows based on pregnancy status. +8 121.60±23.61 50579.21 465.79 108.58 0.000 +1 228.66±13.93-8 158.76±29.79-1 84.91±11.07 total 154.88±57.46 Table 5: Mean serum Total Protein in Holstein dairy cows based on pregnancy status. +8 8.66±0.36 14.98 0.54 27.73 0.000 +1 10.24±0.87-8 9.18±0.43-1 7.70±1.04 total 9.05±1.16 Table 6: correlation index between and urea serum values. urea R= 0.71 Table 7: correlation index between and BHBA serum values. BHBA R= 0.61 Relationship Between and Cholesterol: Based on table 8 and pearson's Correlation index revealed that between and Cholesterol serum values there is a reverse correlation so that correlation index was r= -0.75. This index indicates a reverse effect of on Cholesterol serum values. Thus, with elevating of serum values the Cholesterol values decreased. Relationship Between and Total Protein: Based on table 8 and pearson's Correlation index revealed that between and Total Protein serum values there is a reverse correlation so that correlation index was r= -0.65. This index indicates a reverse effect of on Total Protein serum values. Thus, with elevating of serum values the Total Protein values diminished.

1797 Table 8: Correlation index between and Cholesterol serum values. Cholesterol R= -0.75 Table 8: Correlation index between and Total Protein serum values. Total Protein R= -0.65 Discussion: For awareness of fatty liver syndrome, blood biochemical parameters can be used or we can measure TAG and total fat of hepatic cell. Some researchers inspect fatty liver based on TAB or hepatic fat percent [18]. Raid (1980) divided livers in 4 levels depending on severity of fat accumulation in it: Normal, slight, average and severe [16]. Nowadays general opinion is that a high percent of mature cows show signs of slight or severe fatty liver around parturition [4,11]. Almost near parturition increases in blood and moves to liver, and can cause ketosis, abomasums displacement, metritis and fatty liver after parturition [4,5,8]. In a normal situation and positive energy balance, value is about 200 meq/lit in blood. This value increases since 3 weeks is parturition and reaches to 300 meq/lit in the last week. In the last days before parturition, it reaches to 800-1200mcq/lit. After parturition these acids should wane immediately and if it remains more than 700meq/lit after 7 days, represents negative energy balance and probability of fatty liver incidence. 3 weeks after parturition the amount of these acids should return to normal level (200meq/lit) [5]. Also the results of this study have conformity with Grummer results that showed three is most lipid aggregation in liver in first 4 weeks after parturition [10]. There was a research in Netherlands about 71 dairy cattle before parturition that showed 5 percent of liver is occupied with TAG [12]. Also in a slaughterhouse research in Tehran, aggregation of TAG more than 10% in liver in last month of pregnancy was reported. These researchers had not measured values. In this study, TAG aggregation in liver in last month of pregnancy had occupied more than 5% of liver cells and amount of was more than 900meq/lit being nonspecific and some other reasons. Slight and Mild forms of fatty liver con destroy hepatocytes and disturb liver function without making any changes in activity of hepatic specific enzymes found in serum. Measurement of liver enzymes in serum is useful for evaluating fatty liver disease but with certain restrictions such as is being nonspecific. Mild and moderate forms of fatty liver with damaged hepatocytes can cause liver and no specific changes in liver enzymes in serum, liver dysfunction to establish [2,17]. References 1. Bobe, G., J.W. Young and D.C. Beitz, 2004. Pathology, Etiology, Prevention, and treatment of fatty liver in dairy cows Journal of Dairy Science, 87(10): 3105-3124. 2. Bogin, E., Y. Avidan, M. Merom, S. Soback and G. Brenner, 1988. Biochemical changes associated with the fatty liver syndrome in cows. Journal of Comparative pathology, 98: 337-347. 3. Bruss, M.L., 1993. Metabolic fatty liver of ruminants. In " Advances in veterinary science and comparative medicine, Animal Models in liver research " edited by C.E. Cornelius, 37: 417-449. 4. Drackley, J.K., 1999. Biology of dairy cows during the transition period. Journal of Dairy Science, 82: 2259-2273. 5. Drackley, J.K., 2000. Use of as a Tool to monitor energy balance in transition dairy cows, w.w.w.livestocktrail.uiuc.cdu.uploods/dairy net/, pp: 1-3. 6. Eddy, R.G., 1992. Fatty liver syndrome. In " Bovine Medicin" edited by A.H. Andrews, R.W. Blowey, H. Boyd and R.G. Eddy. Black well scientific publications, London, pp: 598-600. 7. Edmonson, A.J., I.J. Lean, L.D. Weaver, T. Farver and G. Webster, 1989. A body condition scoring chart for Holstein dairy cows.journal of Dairy Research, 72: 68-78. 8. Geelen, M.J.H. and T. Wensing, 2006. Studies on hepatic lipidosis and coinciding health and fertility problems of high-producing dairy cows using the "Utrecht fatty liver model of dairy cows". Veterinary Quarterly, 28(3): 90-104. 9. Goff, J.P. and R.L. Horst, 1997. Physiological changes at parturition and their relationship to metabolic disorders. Journal of Dairy Science, 80: 1260-1268. 10. Grummer, R.R., 1993. Etiology of lipid-related metabolic disorders in periparturient dairy cows. Journal of Dairy Science, 76: 3882-3896. 11. Grummer, R.R., 1995. Impact of changes in organic nutrient metabolism on feeding the transition dairy cow. Journal of Animal Science, 73: 2820-2833.

12. Johannsen, U., S. Menger, R. Staufenbiel and N. Rossow, 1993. Investigations on morphology and function of the liver of high-yielding cows two weeks post partum. Dtsch. Tieraerztl. Wochenschr, 100: 177-181. 13. Jorritsma, R., H. Jorritsma, Y.H. Schukken and G.H. Wentink, 2000. Relationships between fatty liver and fertility and some periparturient diseases in commercial Dutch dairy herds. Theriogenology, 54: 1065-1074. 14. Jorritsma, R., H. Jorritsma, Y.H. Schukken, P.C. Bartlett, T. Wensing and G.H. Wentink, 2001. Prevalence and indicators of postpartum fatty infiltration of the liver in nine commercial dairy herds in the Netherlands. Livest. Product. Science, 68: 53-60. 15. Mcnamara, J.P., 2000. Integrating genotype and nutrition on utilization of body reserves during lactation of dairy cattle. Pages 353-370 in Symposium on Ruminant Physiology. P. B. Cronje, ed. CAB Int., London, UK. 1798 16. Reid, I.M., 1980. Incidence and severity of fatty liver in dairy cows. Veterinary Record, 107: 281-284. 17. Rukkwamsuk, T., T.A.M. Kiuip and T. Wensing, 1999. Relationship between over feeding conditioning in the dairy Peroid and the problems of high producing dairy cows during the post parturient period. Veterinary Quart, 21: 71-77. 18. Wensing, T., T. Kruip, M.J.H. Geelen, G.H. Wentink and A.M. Van den Top, 1997. Postpartum fatty liver in high-producing dairy cows in practice and in animal studies. The connection with health, production and reproduction problems. Comp. Haematol. Int., 7: 167-171. 19. Zerbe, H., N. Schneider, W. Leibold, T. Wensing, T.A.M. Kruip and H.J. Schuberth, 2000. Altered functional and immunophenotypical properties of neutrophilic granulocytes in postpartum cows associated with fatty liver. Theriogenology, 54: 771-786.