Fatty Liver Cows Research Articles

Serum macroelements and microelements levels in periparturient dairy cows in relation to fatty liver diseases

BackgroundFatty liver in dairy cows is a common metabolic disease defined by triglyceride (TG) buildup in the hepatocyte. Clinical diagnosis of fatty liver is usually done by liver biopsy, causing considerable economic losses in the dairy industry owing to the lack of more effective diagnostic methods. Therefore, this study aimed to investigate the potential utility of blood biomarkers for the diagnosis and early warning of fatty liver in dairy cows.ResultsA total of twenty-four lactating cows within 28 days after parturition were randomly selected as experimental animals and divided into healthy cows (liver biopsy tested, n = 12) and cows with fatty liver (liver biopsy tested, n = 12). Inductively coupled plasma mass spectrometry (ICP-MS) was used to determine the macroelements and microelements in the serum of two groups of cows. Compared to healthy cows (C), concentrations of calcium (Ca), potassium (K), magnesium (Mg), strontium (Sr), selenium (Se), manganese (Mn), boron (B) and molybdenum (Mo) were lower and copper (Cu) was higher in fatty liver cows (F). Meanwhile, the observed differences in macroelements and microelements were related to delivery time, with the greatest major disparity between C and F occurring 7 days after delivery. Multivariable analysis was used to test the correlation between nine serum macroelements, microelements and fatty liver. Based on variable importance projection and receiver operating characteristic (ROC) curve analysis, minerals Ca, Se, K, B and Mo were screened as the best diagnostic indicators of fatty liver in postpartum cows.ConclusionsOur data suggested that serum levels of Ca, K, Mg, Se, B, Mo, Mn, and Sr were lower in F than in C. The most suitable period for an early-warning identification of fatty liver in cows was 7 days after delivery, and Ca, Se, K, B and Mo were the best diagnostic indicators of fatty liver in postpartum cows.

Expression of hepatic genes involved in bile acid metabolism in dairy cows with fatty liver

Bile acids are cholesterol-derived molecules that are primarily produced in the liver. In nonruminants with fatty liver, overproduction of bile acids is associated with liver injury. During the transition period, fatty liver is a metabolic disorder that can affect up to 50% of high-producing dairy cows. The purpose of this study was to provide a comprehensive evaluation on hepatic bile acid metabolism in dairy cows with fatty liver by assessing expression changes of genes involved in bile acid synthesis, export and uptake. The serum activities of aspartate aminotransferase, alanine aminotransferase and glutamate dehydrogenase and concentration of total bile acids were all greater, whereas serum concentration of total cholesterol was lower in cows with fatty liver than in healthy cows. Content of total bile acids was higher but total cholesterol was slightly lower in liver tissues from fatty liver cows than from healthy cows. The hepatic mRNA abundance of cholesterol 7a-hydroxylase (CYP7A1), hydroxy-delta-5-steroid dehydrogenase, 3 β- and steroid delta-isomerase 7 (HSD3B7) and sterol 12α-hydroxylase (CYP8B1), enzymes involved in the classic pathway of bile acid synthesis, was higher in fatty liver cows than in healthy cows. Compared with healthy cows, the hepatic mRNA abundance of alternative bile acid synthesis pathway-related genes sterol 27-hydroxylase (CYP27A1) and oxysterol 7α-hydroxylase (CYP7B1) did not differ in cows with fatty liver. The protein and mRNA abundance of bile acid transporter bile salt efflux pump (BSEP) were lower in the liver of dairy cow with fatty liver. Compared with healthy cows, the hepatic mRNA abundance of bile acid transporters solute carrier family 51 subunit α (SLC51A), ATP binding cassette subfamily C member 1 (ABCC1) and 3 (ABCC3) was greater in cows with fatty liver, whereas the solute carrier family 51 subunit β (SLC51B) did not differ. The expression of genes involved in bile acid uptake, including solute carrier family 10 member 1 (NTCP), solute carrier organic anion transporter family member 1A2 (SLCO1A2) and 2B1 (SLCO2B1) was upregulated in dairy cows with fatty liver. Furthermore, the hepatic protein and mRNA abundance of bile acid metabolism regulators farnesoid X receptor (FXR) and small heterodimer partner (SHP) were lower in cows with fatty liver than in healthy cows. Overall, these data suggest that inhibition of FXR signaling pathway may lead to the increased bile acid synthesis and uptake and decreased secretion of bile acids from hepatocytes to the bile, which elevates hepatic bile acids content in dairy cows with fatty liver. As the hepatotoxicity of bile acids has been demonstrated on nonruminant hepatocytes, it is likely that the liver injury is induced by increased hepatic bile acids content in dairy cows with fatty liver.

Evaluation of hepatic AMPK, mTORC1, and autophagy-lysosomal pathway in cows with mild or moderate fatty liver

The aim of the present study was to investigate the activity of AMPK and mTORC1 as well as TFEB transcriptional activity and autophagy-lysosomal function in the liver of dairy cows with mild fatty liver (FL) and cows with moderate FL. Liver and blood samples were collected from healthy dairy cows (n = 10; hepatic triglyceride content <1% wet weight) and cows with mild FL (n = 10; 1% ≤ hepatic triglyceride content < 5% wet weight) or moderate FL (n = 10; 5% ≤ hepatic triglyceride content < 10% wet weight) that had a similar number of lactations (median = 3, range = 2-4) and days in milk (median = 6 d, range = 3-9). Blood parameters were determined using a Hitachi 3130 autoanalyzer with commercially available kits. Protein and mRNA abundances were determined using western blotting and quantitative real-time PCR, respectively. Activities of calcineurin and β-N-acetylglucosaminidase were measured with commercial assay kits. Data were analyzed using one-way ANOVA with subsequent Bonferroni correction. Blood concentrations of glucose were lower in moderate FL cows (3.03 ± 0.21 mM) than in healthy (3.71 ± 0.14 mM) and mild FL cows (3.76 ± 0.14 mM). Blood concentrations of β-hydroxybutyrate (BHB, 1.37 ± 0.15 mM in mild FL, 1.88 ± 0.17 mM in moderate FL) and free fatty acids (FFA, 0.69 ± 0.05 mM in mild FL, 0.96 ± 0.09 mM in moderate FL) were greater in FL cows than in healthy cows (BHB, 0.76 ± 0.12 mM; FFA, 0.42 ± 0.04 mM). Compared with healthy cows, phosphorylation of AMPK was greater and phosphorylation of its downstream target acetyl-CoA carboxylase 1 was lower in cows with mild and moderate FL. Phosphorylation of mTOR was lower in cows with mild FL compared with healthy cows. In cows with moderate FL, phosphorylation of mTOR and its downstream effectors was greater than in healthy cows and cows with mild FL. The mRNA abundance of TFEB was downregulated in cows with moderate FL compared with healthy cows and mild FL cows. In mild FL cows, the mRNA and protein abundances of TFEB were greater than in healthy cows. Compared with healthy cows, the mRNA abundances of autophagy markers sequestosome-1 and microtubule-associated protein 1 light chain 3-II, and the protein and mRNA abundances of lysosome-associated membrane protein 1 and cathepsin D were increased in mild FL cows but decreased in moderate FL cows. Compared with healthy cows, the mRNA abundance of mucolipin 1 and activities of β-N-acetylglucosaminidase and calcineurin were higher in cows with mild FL but lower in cows with moderate FL. These data demonstrate that hepatic AMPK signaling pathway, TFEB transcriptional activity, and autophagy-lysosomal function are increased in dairy cows with mild FL; the hepatic mTORC1 signaling pathway is inhibited in mild FL cows but activated in moderate FL cows; and activities of AMPK and TFEB as well as autophagy-lysosomal function are impaired in moderate FL cows.

Transcriptome analysis reveals disruption of circadian rhythms in late gestation dairy cows may increase risk for fatty liver and reduced mammary remodeling.

Circadian disruption increased insulin resistance and decreased mammary development in late gestation, nonlactating (dry) cows. The objective was to measure the effect of circadian disruption on transcriptomes of the liver and mammary gland. At 35 days before expected calving (BEC), multiparous dry cows were assigned to either control (CON) or phase-shifted treatments (PS). CON was exposed to 16-h light and 8-h dark. PS was exposed to 16-h light to 8-h dark, but phase of the light-dark cycle was shifted 6 h every 3 days. On day 21 BEC, liver and mammary were biopsied. RNA was isolated (n = 6 CON, n = 6 PS per tissue), and libraries were prepared and sequenced using paired-end reads. Reads mapping to bovine genome averaged 27 ± 2 million and aligned to 14,222 protein-coding genes in liver and 15,480 in mammary analysis. In the liver, 834 genes, and in the mammary gland, 862 genes were different (nominal P < 0.05) between PS and CON. In the liver, genes upregulated in PS functioned in cholesterol biosynthesis, endoplasmic reticulum stress, wound healing, and inflammation. Genes downregulated in liver function in cholesterol efflux. In the mammary gland, genes upregulated functioned in mRNA processing and transcription and downregulated genes encoded extracellular matrix proteins and proteases, cathepsins and lysosomal proteases, lipid transporters, and regulated oxidative phosphorylation. Increased cholesterol synthesis and decreased efflux suggest that circadian disruption potentially increases the risk of fatty liver in cows. Decreased remodeling and lipid transport in mammary may decrease milk production capacity during lactation.

Effect of heat stress on the values of biochemical blood parameters of dairy cattle

The aim of the study was to determine the effect of heat stress on differences between selected biochemical blood parameters in cows in June and October, taking into account different housing systems and cooling systems used. The research was carried out on 60 cows from stanchion barns (farm A) and loose-house barns (farm B). Material for research, venous blood, was collected at the same time, in the morning, after milking and clinical examination. Then, parameters describing the energy balance and the activity indexes of the parenchymal organs as well as the mineral balance were determined. Milk production in both farms was clearly lower in June than in October, but the difference between milk yields obtained from the two farms was over 15% in June and only less than 10% (5-6 kg of milk) in October. Both farms A and B showed an increase in aspartate aminotransferase and creatine kinase. In farm A, the statistically significant increase exceeded the normal physiological range, which could indicate developing disturbances in the locomotor system caused by the impact of high temperature. Energy balance disturbances occurred only in farm A, where a statistically significant decrease in glucose and cholesterol concentration was found in June samples, which, according to the authors, could be related to a higher energy demand. The results of biochemical tests of venous blood collected in June may indicate the onset of fatty liver in cows (decreased cholesterol, glucose, and total protein levels, as well as increased bilirubin levels and GGTP activity) as a consequence of exposure to high temperatures. Heat stress has a significant impact on the functioning of the cow’s organism, which will not always be manifested by characteristic clinical signs. Accurate laboratory analyses make it possible to diagnose changes developing as a result of cows being exposed to heat stress.

Low Expression of Sirtuin 1 in the Dairy Cows with Mild Fatty Liver Alters Hepatic Lipid Metabolism.

Simple SummarySirtuin 1 (SIRT1), a NAD-dependent histone deacetylase, is involved in oxidative stress and lipid metabolism regulation. Limited studies exist regarding the role of SIRT1 in lipid metabolism disorder in periparturient dairy cows. This study explores the effect of hepatic steatosis on the expression of the SIRT1 gene and protein and the proteins encoded by the genes downstream to it, all of which are involved in lipid metabolism in the liver. Control cows (n = 6, parity 3.0 ± 2.0, milk production 28 ± 47 kg/d) and mild fatty liver cows (n = 6, parity 2.3 ± 1.5, milk production 20 ± 6 kg/d) were retrospectively selected based on liver triglycerides (TG) content (% wet liver). The present study indicates that low SIRT1 expression caused by hepatic steatosis promotes hepatic fatty acid synthesis and inhibits fatty acid β-oxidation. We believe that our study makes a significant contribution to the literature because it demonstrates that hepatic steatosis is associated with increased hepatic fatty acid synthesis, inhibited fatty acid β-oxidation and reduced lipid transport.Dairy cows usually experience negative energy balance coupled with an increased incidence of fatty liver during the periparturient period. The purpose of this study was to investigate the effect of hepatic steatosis on the expression of the sirtuin 1 (SIRT1), along with the target mRNA and protein expressions and activities related to lipid metabolism in liver tissue. Control cows (n = 6, parity 3.0 ± 2.0, milk production 28 ± 7 kg/d) and mild fatty liver cows (n = 6, parity 2.3 ± 1.5, milk production 20 ± 6 kg/d) were retrospectively selected based on liver triglycerides (TG) content (% wet liver). Compared with the control group, fatty liver cows had greater concentrations of cholesterol and TG along with the typically vacuolated appearance and greater lipid droplets in the liver. Furthermore, fatty liver cows had greater mRNA and protein abundance related to hepatic lipid synthesis proteins sterol regulatory element binding proteins (SREBP-1c), long-chain acyl-CoA synthetase (ACSL), acyl-CoA carbrolase (ACC) and fatty acid synthase (FAS) and lipid transport proteins Liver fatty acid binding protein (L-FABP), apolipoprotein E (ApoE), low density lipoprotein receptor (LDLR) and microsomal TG transfer protein (MTTP) (p < 0.05). However, they had lower mRNA and protein abundance associated with fatty acid β-oxidation proteins SIRT1, peroxisome proliferator-activated receptor co-activator-1 (PGC-1α), peroxisome proliferator–activated receptor-α (PPARα), retinoid X receptor (RXRα), acyl-CoA 1 (ACO), carnitine palmitoyltransferase 1 (CPT1), carnitine palmitoyltransferase 2 (CPT2) and long- and medium-chain 3-hydroxyacyl-CoA dehydrogenases (LCAD) (p < 0.05). Additionally, mRNA abundance and enzyme activity of enzymes copper/zinc superoxide dismutase (Cu/Zn SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and manganese superoxide dismutase (Mn SOD) decreased and mRNA and protein abundance of p45 nuclear factor-erythroid 2 (p45 NF-E2)-related factor 1 (Nrf1), mitochondrial transcription factor A (TFAM) decreased (p < 0.05). Lower enzyme activities of SIRT1, PGC-1α, Cu/Zn SOD, CAT, GSH-Px, SREBP-1c and Mn SOD (p < 0.05) and concentration of reactive oxygen species (ROS) were observed in dairy cows with fatty liver. These results demonstrate that decreased SIRT1 associated with hepatic steatosis promotes hepatic fatty acid synthesis and inhibits fatty acid β-oxidation. Hence, SIRT1 may represent a novel therapeutic target for the treatment of the fatty liver disease in dairy cows.

Short communication: Enhanced autophagy activity in liver tissue of dairy cows with mild fatty liver

During the transition period, dairy cows are challenged by increased energy demands and decreased dry matter intake, which can induce a variety of metabolic disorders, especially fatty liver. Dairy cows suffering from mild or moderate fatty liver in this period show no distinct clinical symptoms, indicating the occurrence of adaptive processes. The process of autophagy (an adaptive response) leads to degradation of intracellular components to generate energy and maintains cellular homeostasis during negative nutrient status. Whether autophagy is involved in metabolic adaptations of the pathological course of mild fatty liver is unclear. Thus, the aim of this study was to determine hepatic autophagy status in dairy cows with mild fatty liver. Liver samples were collected from healthy cows (n = 15), defined as having hepatic triglyceride (TG) content <1% on a wet weight basis, and cows with mild fatty liver (n = 15), defined as having hepatic TG content between 1 and 5%. The abundance of the ubiquitinated proteins, microtubule-associated protein 1 light chain 3 (MAP1LC3, also called LC3-II) and sequestosome-1 (SQSTM1, also called p62) was lower, whereas the mRNA abundance of MAP1LC3 and SQSTM1 was greater in cows with mild fatty liver. The hepatic mRNA abundance of autophagy-related (ATG) genes ATG5 and ATG7 was greater in response to fatty liver. However, the protein abundance of ATG5 and ATG7 did not differ between healthy and mild fatty liver cows. Together, these data indicate that the formation and degradation of autophagosomes is enhanced in the liver of cows with mild fatty liver. Besides, these results are conducive to define the adaptation mechanisms of dairy cows during the transition period.

Fatty acid-induced endoplasmic reticulum stress promoted lipid accumulation in calf hepatocytes, and endoplasmic reticulum stress existed in the liver of severe fatty liver cows

Disruption of endoplasmic reticulum (ER) homeostasis, often termed ER stress, is intrinsically linked with perturbation of lipid metabolism in humans and mice. Whether ER homeostasis is affected in cows experiencing fatty liver is unknown. The aim of this study was to investigate the potential role of ER stress in hepatic lipid accumulation in calf hepatocytes and ER stress status in dairy cows with severe fatty liver. In vitro experiments were conducted in which hepatocytes were isolated from calves and treated with different concentrations of fatty acids, tauroursodeoxycholic acid (TUDCA; a canonical inhibitor of ER stress), or both. The increase in phosphorylation level of protein kinase RNA-like ER kinase (PERK) and inositol requiring protein-1α (IRE1α) proteins, and the cleavage of activating transcription factor-6 (ATF6) protein in response to increasing doses of fatty acids (which were reversed by TUDCA treatment) in primary hepatocytes underscored a mechanistic link between fatty acids and ER stress. In addition, fatty acid treatment increased the abundance of sterol regulatory element-binding protein 1c, acetyl-CoA carboxylase-α, fatty acid synthase, and diacylglycerol acyltransferase 1, and lipid accumulation in calf primary hepatocytes, whereas inhibition of ER stress by incubating with TUDCA significantly weakened these effects. Overall, results in vitro indicate that inhibition of ER stress in calf hepatocytes alleviates fatty acid-induced lipid accumulation by downregulating the expression of lipogenic genes. In vivo experiments, liver and blood samples were collected from cows diagnosed as healthy (n = 15) or with severe fatty liver (n = 15). The phosphorylation level of PERK and IRE1α, the cleavage of ATF6 protein, and the abundance of several unfolded protein response genes (78 kDa glucose-regulated protein, AMP-dependent transcription factor 4, and spliced X-box binding protein 1) were greater in liver of cows with severe fatty liver. The present in vivo study confirms the occurrence of ER stress in dairy cows with severe fatty liver. Considering the causative role of fatty acid-induced ER stress in hepatic lipid accumulation in calf hepatocytes, the existence of ER stress in the liver of severe fatty liver cows may presage its participation in fatty liver progression in dairy cows. However, the mechanistic relationship between ER stress and fatty liver in dairy cows remain to be determined.

Exploration of serum sensitive biomarkers of fatty liver in dairy cows

Serum proteins are sensitive with diseases in dairy cows, and some of them could be used as biomarkers for fatty liver. This study aimed to explore serum biomarkers for fatty liver in dairy cows. A total of 28 early lactating dairy cows were chosen from a commercial dairy herds, liver samples were collected for determining concentration of triacylglycerol (TAG), and serum samples were collected for measuring fibroblast growth factor-21 (FGF-21), adiponectin, Lipoprotein-associated phospholipase A2 (LP-PLA2), and hemoglobin (Hb). Dairy cows were divided into fatty liver (liver TAG > 5%, wet weight) and control group (liver TAG < 5%, wet weight). Concentration of FGF-21 was greater in fatty liver cows, while the concentration of LP-PLA2 and Hb was less. The concentration of FGF-21 and total Hb had strong correlation with the liver TAG as well as good prediction power (kappa value = 0.79 and 0.58, respectively). These results suggested that the serum concentration of FGF-21 and total Hb could be potentially used as fatty liver biomarkers in lactating dairy cows.

Serum Ornithine Carbamoyltransferase Activity and Correlation with Fatty Liver in Dairy Cows with Displaced Abomasum

Ornithine Carbamoyltransferase (OCT) is a hepatocyte-specific enzyme; elevated serum activity indicates liver damage. Displaced abomasum frequently occurs around the time of parturition in dairy cattle and is associated with fatty liver. We analyzed serum OCT activities to investigate the relationship between displaced abomasum and fatty liver. Lactating Holstein cows, 75 clinically healthy and 22 with displaced abomasum, were used. Serum OCT activities were compared between healthy cows and those with displaced abomasum. Liver biopsies of cows with displaced abomasum were classified by the severity of fat accumulation. Blood samples were collected preoperatively and at 4 and 8 days after surgery and analyzed for serum OCT, aspartate aminotransferase, alkaline phosphatase, γ-glutamyltransferase, urea nitrogen, total protein and hematocrit. Serum OCT activity was significantly higher in cows with displaced abomasumthan in healthy cows. The degree of fatty liver in cows with displaced abomasum was classified as mild (13 cows), moderate (5 cows), or severe (4 cows). Statistical analysis revealed no significant differences in blood test results among cows with different degrees of fatty liver. However, our results indicated significant differences in serum OCT, aspartate aminotransferase, urea nitrogen, total protein and hematocrit when stratified by the number of days after surgical treatment for displaced abomasum. Serum OCT activity is increased in dairy cows with displaced abomasum and the influence of the degree of fatty liver on serum OCT activity is unclear. Therefore, OCT should be reconsidered as a marker of fatty liver in cows around the time of parturition, when displaced abomasum frequently occurs. Fatty liver should be evaluated by various clinical parameters, not by one hepatic enzyme.

New insights into body condition score and its association with fatty liver in holstein dairy cows

The aim of this study was to estimate the association between body condition and fatty liver in high-yielding dairy cows. One hundred dry Holstein cows were selected. Cows were scored once for body condition during the dry period, puerperium and month 2 of lactation, according to the system provided by Elanco Animal Health Buletin Al 8478. Body condition loss was determined as marked if loss was over 0.7 points between two consecutive phases of cycles and over 1.5 points between puerperium and month 2 of lactation. Liver tissue samples were taken 12 d after calving and tested for lipid content. 50% of cows had mild (3.92 ± 3.33% fat), 33% moderate (19.28 ± 5.18% fat), and 17% severe fatty liver (36.21 ± 4.55% fat). The mean body condition scores were 3.79 ± 0.55 (dry period), 3.18 ± 0.34 (puerperium) and 2.90 ± 0.29 (month 2 of lactation). Marked body condition loss from the dry period to puerperium had 38% of cows, 20% from dry period to month 2 of lactation and 8% from puerperium to month 2 of lactation. Body condition scores out of the physiological range had 39% dry cows, 49 % cows in puerperium and 9% cows at month 2 of lactation. After dividing cows into groups according to fatty liver degree, it was shown that only cows with severe fatty liver were obese during the dry period and that 76.47% of cows from this group had marked condition loss from the dry period to puerperium, 47.06% from dry period to month 2 of lactation and 23.53% from puerperium to month 2 of lactation. These indicate that in cows with severe fatty liver lipomobilisation is intensive and starts around calving, before milk production enhacement. Our results indicate a strong association between body condition and fatty liver in cows, which can be estimated only when body condition is analyzed in details. Besides, our results clearly indicate that body condition is not the only etiological factor that leads to fatty liver in dairy cows.

Possibilities for preventing fatty liver in cows during peripartal period

The objective of these investigations was to examine possibilities for preventing fatty liver in cows during the peripartal period using the preparation HEPARENOL which includes methionine, lysine and choline in its composition. Investigations were performed on cattle of the Holstein breed whose average production during their last lactation was 8120 liters. The cows included in the experiment were divided into three groups of 10 cows each. Cows of the first group were administered the preparation HEPARENOL perorally 5-7 days before partus in daily doses of 50 ml per animal, and then daily during the first seven days after parturition in daily doses of 100 ml per animal. The second group of cows were administered the preparation in the same way as the first group, but they also received vitamin C injected deep i.m. in doses of 1000 mg per animal per day. The third group of cows were not exposed to any treatment and served as the control group. The body condition of the cows was marked according to the system Elanco Animal Health Bulletin Al 8478. Blood samples were taken by puncture of the jugular vein before the beginning of treatment, and on the third and 12th days after partus. The glucose concentration was determined in the blood samples. On the 12th day after partus, liver tissue samples were taken by biopsy for pathohistological examinations. Since the cows were fed in the same way and were in the same phase of the production-reproduction cycle, the average score of the body condition before treatment was approximately the same. However, on the 3rd day after partus, cows of the second experimental group showed average values that were significantly lower in comparison with the other two groups of cows. It is very interesting that a significantly lower value of glycemia (x=1.93?0.34 mmol/l) was established in the same group of cows at this time. In cows of the first experimental and the control group, glycemia was approximately the same during the examined period. In cows of the second experimental group on the 12th day after partus the glucose concentration significantly increased (x=2.42?0.58 mmol/l) against the values obtained for the 3rd day, but it was still significantly lower in comparison with the glycemia of cows of the first experimental (x=3.34?0.61 mmol/l) and the control group (x=3.48?0.49 mmol/l). Practically, throughout the entire period glycemia was statistically significantly lower in the second experimental group in comparison with the other two groups of cows. The degree of fatty liver on the 12th day postpartally, or five days after the end of treatment, was the lowest in cows of the second experimental group. In order to stress the differences, it is pointed out that there were two cows without fatty liver in the second group, while there were no such cows in the first experimental or in the control group. This group contained the smallest number of cows with a mean degree of fatty liver (n=2), while two cows had very strong fatty liver. There were no differences between the control and the first experimental group in the number of cows with a mean degree of fatty liver, with the exception that the control group had two and the first experimental group one cow with a mild degree of fatty liver. It seems that treatment using the preparation HEPARENOL, or methionine, lysine and choline, before and after parturition did not have a significant effect on the incidence and degree of fatty liver. Only cows with additional and simultaneous treatment with vitamin C had a lower incidence and degree of morphological changes in the liver, in addition to the sigfnificant changes in body condition and the blood glucose concentration.

Decrease in stearic acid proportions in adipose tissues and liver lipids in fatty liver of dairy cows

ABSTRACTSamples of liver and perirenal, mesenteric and subcutaneous fat were collected from 16 sick necropsied dairy cows to evaluate the fatty acid profiles in the hepatic and adipose tissues associated with advanced fatty liver or hepatic lipidosis. Hepatic triglyceride and eight fatty acids were measured in the hepatic and adipose tissues. Six cows had more than 3% triglyceride on fresh weight in their livers and were classified as having fatty liver. Stearic and linoleic acid proportions in the liver decreased markedly with increased hepatic triglyceride levels, while the proportion of palmitic and oleic acids increased. The most striking fluctuations in hepatic lipidosis were manifested as decreased stearic acid in the adipose tissues including subcutaneous fat with the trend of decreasing stearic acid. Palmitic acid was elevated in hepatic and perirenal fat in fatty liver cows. In instances of advanced hepatic lipidosis, palmitoleic acid increased in only subcutaneous fat and not in perirenal or mesenteric fat. In addition to the proportions of hepatic fatty acids in fatty liver, this study also clarified the fluctuations observed in the profiles of fatty acids of the adipose tissues in cows with advanced hepatic lipidosis, particularly the decline in the proportions of stearic acid.

Strong relationships between mediators of the acute phase response and fatty liver in dairy cows

The objective of this study was to investigate the relationship between activation of acute phase response and fatty liver in transition dairy cows. Fatty liver was induced in dairy cows by feeding 8 kg of cracked corn 1 mo before the expected day of parturition. Liver and blood samples were obtained at days -4, 3, 8, 12, 14, 22, 27, and 36 postpartum. Cows that developed fatty liver (n = 4) reached peak total lipids in the liver at day 12 postpartum with 11.4% (wet wt.) compared with 6.6% in control cows (n = 4). Cows with fatty liver had greater plasma tumor necrosis factor-alpha (TNF-α), nonesterified fatty acids (NEFA), and lower lactate concentrations than did control cows at day -4. During highest concentrations of total lipids in the liver, for at least one time-point, fatty-liver cows had greater concentrations of plasma serum amyloid A (SAA), haptoglobin, and NEFA and lower concentrations of calcitonin gene-related peptide (CGRP), prostaglandin E2 (PGE2), cortisol, and TNF-α than did control cows. Concentrations of total lipids in the liver at 12 d postpartum were correlated (a) positively with plasma TNF-α, SAA, and NEFA and negatively with plasma CGRP before calving, (b) positively with plasma SAA, haptoglobin, and NEFA and negatively with plasma PGE2, CGRP, total cholesterol, and glucose at days 3, 8, and 12 postpartum, and (c) negatively with concentrations of plasma glucose, lactate, and total bilirubin after day 12 postpartum. In conclusion, this study indicates that the acute phase response occurs in cows with fatty liver as well as strong relationships between mediators of immune response and fatty liver. Key words: Acute phase response, fatty liver, dairy cow

Fatty acid profiles in relation to triglyceride level in the liver of dairy cows.

To elucidate possible relationships between triglyceride (TG) levels and fatty acid composition in bovine liver, hepatic TG and seven individual fatty acids were measured in 23 Holstein dairy cows, of them 6 are healthy. Liver TG level was greater than 3 % in 12 cows which were ruled fatty liver. Palmitic and oleic acid proportions were significantly higher in fatty liver cows than in the healthy cows, while stearic acid was lower in fatty liver cows. With increased liver TG, stearic acid proportions decreased dramatically. Results indicate that hepatic lipidosis markedly alters the proportions of the various fatty acids in the liver of dairy cows.

Decreases of apolipoprotein B-100 and A-I concentrations and induction of haptoglobin and serum amyloid A in nonfed calves.

Reduced feed intake near parturition is suggested to be one of the major causal factors for the development of fatty liver in cows, and nonfeeding has been used as an experimental model for fatty liver. In cows with fatty liver, concentrations of lipoprotein lipids and proteins are decreased. In addition, the acute-phase protein haptoglobin is induced. The purpose of the present study was to examine whether the decrease of lipoprotein concentrations and the induction of acute-phase proteins were similarly reproduced by non-feeding. Holstein female calves (n=5) were nonfed for 3 days and thereafter refed. Serum concentrations of nonesterified fatty acids and beta-hydroxybutyric acid were initially increased by the nonfeeding, and followed by decreases in concentrations of cholesteryl esters, phospholipids, apolipoprotein (apo) B-100 and apoA-I. The apoC-III concentration was not distinctly decreased. Haptoglobin and serum amyloid A were induced during the nonfeeding and refeeding process. Haptoglobin was distributed in different proportions in the high-density lipoprotein, very high-density lipoprotein and the lipoprotein-deficient fractions, whereas almost all serum amyloid A was associated with the high-density lipoprotein fraction. These results suggest that the decreases in lipoprotein concentrations and induction of acute-phase proteins found in cows with fatty liver and those with fatty liver-related diseases such as ketosis are primarily due to the reduced feed intake near parturition.

Glucose and insulin responses to glucagon injection in dairy cows with ketosis and fatty liver.

The purpose of this investigation was to study the metabolic situation in clinical cases of bovine ketosis and to diagnose additional diseases. Extensive clinical examination, clinical biochemistry, haematology and fine-needle aspiration biopsy of liver was performed on 17 ketotic and eight control dairy cows in the field, and on seven hospitalized hyperketonaemic fatty liver patients. Additional findings in the ketotic group were heat (n = 7), indigestion (n = 5), endometritis (n = 2), cystic ovaries (n = 1), and mastitis (n = 1), and in the fatty liver group displaced abomasum (n = 4), abomasal ulcers (n = 3), mastitis (n = 2), laminitis (n = 1), bronchopneumonia (n = 1), and hypomagnesaemia (n = 2). There were no additional findings in the control group. Aspartate aminotransferase (AST) and creatine kinase (CK) were elevated in the ketosis and fatty liver groups. Total bilirubin, gamma-glutamyl transferase (GGT) and glutamate dehydrogenase (GD) were elevated in the fatty liver group and in some animals in the ketosis group. Total bile acid was not different between the groups. The free fatty acid/cholesterol ratio was higher in the fatty liver group compared with the control and ketosis groups. There was no or only slight fatty degeneration of the liver cells in the control and ketosis groups. Glucose and insulin preinjection concentrations and changes from basal values after glucagon injection were significantly lower in the ketosis group if compared with the control group. The responses in the fatty liver animals after glucagon injection were more heterogeneous than in the control and ketosis animals, a sign of disturbance in the metabolic adaptation, which together with high free fatty acid (FFA) levels can lead to fatty liver in cows with concurrent diseases.

Reduced protein kinase C activity and endogenous protein phosphorylation in ethionine-induced fatty liver in cows.

Protein kinase C (PKC) activity was evaluated and the phosphorylation of its endogenous substrates was explored in fatty liver induced by administration of ethionine (an analogue of methionine) to cows in order to assess the relevance of PKC-dependent phosphorylation in the development of fatty liver. PKC activity was decreased in both the cytosolic and the total particulate fractions from fatty livers, compared to the corresponding fractions from control liver. The mode of activation by the PKC cofactors (1-oleoyl-2-acetyl-sn-glycerol, 12-O-tetradecanoylphorbol-13-acetate, phosphatidyl-serine and Ca2+) was similar in both control and fatty livers, suggesting a quantitative but not a qualitative change in PKC in fatty liver. At least three substrate proteins (34 kDa, 26 kDa and 19 kDa) were found in the cytosolic fraction and their phosphorylation was reduced in fatty liver. These results suggest that impairment of the signal transduction pathway mediated by PKC is involved in the pathogenesis of fatty liver in cows.

Possible involvement of protein kinase C with induction of haptoglobin in cows by treatment with dexamethasone and by starvation

Summary Haptoglobin (Hp), an acute-phase protein, is detected in serum of cows with hepatic lipidosis (fatty liver). To assess the relevance of Hp in fatty liver, induction of Hp was examined, using conditions similar to those involving development of fatty liver in cows. Induction of Hp was achieved by a combination of dexamethasone administration (0.1 mg/kg of body weight) and 2 days’ starvation. Haptoglobin appearance in serum was not associated with the increase of α1-acid glycoprotein (a marker for inflammation). This treatment increased serum nonesterified fatty acids concentration and decreased serum triglycerides concentration. Protein kinase C activity was decreased in the cytosolic fractions of liver and mononuclear cells. Reduction of protein kinase C-catalyzed endogenous protein phosphorylation also was observed, particularly in the cytosolic fractions of the tissue and cells. Detection of Hp in serum of cows with fatty liver appears to be explained by the fact that Hp is induced by dexamethasone administration and starvation, which are similar to the condition responsible for fatty liver development. The change of protein kinase C-catalyzed phosphorylation was suggested to be involved in the induction of Hp in cows.

Studies on the pathogenesis of fatty liver in cows

Increased fat mobilization in early lactation in cows is described by many authors. Energy deficit in connection with the start of milk secretion is considered to be of importance as cause of the lipolysis. One consequence of increased mobilization of body fat can be development of a fatty liver.&#x0D; In an in vitro of lipid metabolism in bovine adipose tissue McNamara and Hillers have shown that there is an adaptation in prepartum. There is a strong reduction in lipogenesis and esterification and increase in norepinephrine- and epinephrine-stimulated free fatty acid release. The lipoprotein lipase activity in adipose tissue decreases and increases in the mammary gland. This metabolic shift precedes the energy demand for lactation and is stimulated by prolactin and inhibited by progesterone. At two months postpartum there is a dramatic rebound in lipogenesis and esterification.&#x0D; Besides the increased transport of fatty acids to the liver near partus, decreased removal of fat from the liver can be of importance in the development of fatty liver. The aim of the present investigation was to elucidate the pathogenesis of fatty liver in cows by analysing serum lipoproteins and plasma lipids in normal cows round partus on different prepartum feeding and in cows with fatty liver.