Plasma lipoproteins and fatty liver in dairy cows

Dairy cows were classified on the basis of a histological study after a hepatic biopsy conducted in the second week post partum (mild fatty liver, moderate fatty liver, severe fatty liver). Plasma lipoproteins were separated into various density classes by repeated ultracentrifugation. The results indicate that the beginning of lactation is associated with a low concentration of the 1.006 to 1.063 g ml-1 lipoprotein fraction. The lowest concentrations occurred in cows with severe steatosis or during the evolution of moderate steatosis.     

Reduced fertility associated with fatty liver in high-yielding dairy cows

In a group of 20 high-yielding dairy cows, 10 with mild and 10 with severe fatty liver one week after calving, there was an association between fatty liver and reduced fertility. The 10 cows with severe fatty liver had a significantly longer calving interval to the next lactation compared to the cows with mild fatty liver. In addition there was a significant positive correlation between the amount of fat in the liver and calving interval. The results support the concept that liver dysfunction at calving adversely affects subsequent fertility.     

奶牛亚临床酮病检测

本研究采用试纸条法、紫外分光光度法和酮粉法检测奶牛亚临床酮病,结果表明：以紫外分光光度法定量检测结果为诊断标准,试纸条法具有较高的敏感性和特异性,分别为69％和94％,酮粉法的检测敏感性和特异性较低,分别为56％和90％.     

Hepatocyte apoptosis in dairy cows with fatty infiltration of the liver

The objective of the present study was to analyse the apoptotic process of liver cells in dairy cows with fatty infiltration of the liver using indicators of DNA damage and immunohistochemistry. For this purpose, sixteen dairy cows with fatty liver were examined. On clinical examination, the physical condition of the animals was fair in nine and poor in seven cows. The most dominant clinical signs were reduced ruminal motility, inappetance and/or anorexia and recumbency. Postmortem examination, in seven cases, revealed enlarged liver (18-33kg), icteric carcasses and distended gallbladder. Laboratory results included neutrophilia, hypochloraemia, decreased concentrations of total bilirubin and increased concentrations of β-hydroxy butyric acid, non-esterified fatty acids and insulin. The activities of aspartate aminotransferase, γ-glutamyl transpeptidase, creatine kinase and lactate dehydrogenase were high. Histopathological examination of hepatic specimen showed lipid drops in cytosol with indistinct cellular membranes. In control hepatic cells, the DNA was tightly compressed and maintained the circular disposition of the normal nucleus. However, in the diseased cows, the damaged DNA migrated from the core toward the anode, forming a tail of a comet. Compared to controls, numerous ssDNA and caspase-3-positive cells were detected in the liver. To the authors' knowledge, this is the first study to document accelerated apoptosis of hepatocytes in dairy cows with fatty infiltration of the liver.     

泌乳奶牛脂肪肝早期诊断的研究

为了寻找泌乳奶牛脂肪肝早期诊断方法,本研究从某千头奶牛场随机选取了30头乳酮阳性的奶牛,其中临床酮病牛10头,亚临床酮病牛20头,测定其肝脂含量以及血中代谢指标和肝功指标。试验结果表明,脂肪肝的奶牛存在能量负平衡现象,机体呈现高游离脂肪酸血症、高酮血症,肝脂浸润越重,能量代谢障碍越严重;乳酮阳性出现时间的早晚与脂肪肝的严重程度密切相关,乳酮阳性产后出现的越早,乳酮阳性越明显,肝脂浸润越重;随肝脂浸润程度的加重,肝功多个指标会出现异常,重度的脂肪肝会引发肝功能不全。本试验证明,酮粉法可以作为早期诊断奶牛脂肪肝的初选方法,肝功指标异常可作为诊断脂肪肝的辅助方法,两者结合可作为奶牛脂肪肝的实用、有效和准确的早期诊断方法,准确率可达80%。     

Hepatocyte apoptosis in dairy cows with fatty infiltration of the liver

The objective of the present study was to analyse the apoptotic process of liver cells in dairy cows with fatty infiltration of the liver using indicators of DNA damage and immunohistochemistry. For this purpose, sixteen dairy cows with fatty liver were examined. On clinical examination, the physical condition of the animals was fair in nine and poor in seven cows. The most dominant clinical signs were reduced ruminal motility, inappetance and/or anorexia and recumbency. Postmortem examination, in seven cases, revealed enlarged liver (18–33 kg), icteric carcasses and distended gallbladder. Laboratory results included neutrophilia, hypochloraemia, decreased concentrations of total bilirubin and increased concentrations of β-hydroxy butyric acid, non-esterified fatty acids and insulin. The activities of aspartate aminotransferase, γ-glutamyl transpeptidase, creatine kinase and lactate dehydrogenase were high. Histopathological examination of hepatic specimen showed lipid drops in cytosol with indistinct cellular membranes. In control hepatic cells, the DNA was tightly compressed and maintained the circular disposition of the normal nucleus. However, in the diseased cows, the damaged DNA migrated from the core toward the anode, forming a tail of a comet. Compared to controls, numerous ssDNA and caspase-3-positive cells were detected in the liver. To the authors’ knowledge, this is the first study to document accelerated apoptosis of hepatocytes in dairy cows with fatty infiltration of the liver.     

奶牛亚临床低血钙症的蛋白质组学分析

根据奶牛血浆钙浓度将奶牛分为试验组和对照组,并设置2个平行试验组。应用iTRAQ/LC-MS/MS技术对2组奶牛样品进行差异表达蛋白的筛选和鉴定,并结合生物信息学软件对试验结果进行分析。试验共得到398个差异表达蛋白,其中2个平行试验组重叠的蛋白为265个,经过筛选共得到14种具有统计学差异的表达蛋白。结合生物信息学分析中的基因功能分析,结果得到74个注释结果,细胞组成、分子功能和生物过程分别为6,12,55个。根据结果分析可知,奶牛亚临床低血钙症生理和病理现象能够从3个方面解释其生理和病理现象,分别是血钙调节、免疫与炎症反应和血液凝固与补体途径。本试验首次应用iTRAQ结合LC-MS/MS技术对亚临床低血钙症奶牛进行蛋白质组学分析,填补该技术对亚临床低血钙症的空白,并提供科学依据。     

Total serum bile acid concentrations in dairy cows with fatty liver and liver failure

In forty-five Holstein Frisian dairy cows (1-6 weeks post partum; mean age: 5.1 +/- 1.2 years) the serum total bile acid concentrations (SBA) were measured enzymatically. In all cows a left sided abomasal displacement was corrected surgically by right side laparotomy and omentopexy three days before investigation. The liver fat content was determined in all cows histologically. Liver failure was assumed if typical clinical signs (ataxia, general depression, recumbency or coma), an increased venous plasma ammonia level (> 35 mumol/l) and a decreased plasma amino acid index (< 4.0) were found. Cows without liver failure (N = 29) were grouped according to the liver fat content as cows with mild (N = 5), moderate (N = 19) or severe hepatosteatosis (N = 5). Histological examination of liver biopsies in cows with liver failure (N = 16) revealed in twelve cases a severe fatty liver and in four cases a hydropic degeneration of the liver tissue. Although in cows without liver failure mean SBA concentrations were higher in the group with moderate (47.3 +/- 30.9 mumol/l) or severe fatty liver (32.9 +/- 21.7 mumol/l) than in that with mild lipidosis (18.0 (16.8 mumol/l), differences were not significant. The mean SBA concentration in cows with liver failure (70.5 +/- 49.5 mumol/l) was only significantly (p < 0.05) increased compared to cows with uncomplicated mild hepatic lipidosis. In conclusion, the determination of SBA concentrations is of little value in the recognition of fatty liver or even liver failure due to the considerable variance of SBA concentrations in dairy cows.     

Efficacy of sodium borate in the prevention of fatty liver in dairy cows

The effects of sodium borate (100 mg/kg body weight, p.o., 15 days) from a month before expected calving until a month after calving were evaluated in dairy cows susceptible to fatty liver. Cows received either sodium borate (n = 13) or no treatment (n = 10). All cows had mild fatty livers and increased plasma triglycerides and very low density lipoprotein (VLDL) concentrations at the beginning of the experiment. The control group of cows developed significant fatty liver after calving, and 2 of them had severe fatty liver associated with clinical and biochemical abnormalities. There were no clinicopathological signs related to sodium borate administration. Serum triglycerides and VLDL concentrations before calving decreased significantly at calving and after calving in controls, and they were within the normal range only after calving. There were significant alterations during the experiment in some hematological and chemical variables between groups, within period, but they were within the normal range. Unlike treated cows, serum triglycerides and VLDL concentrations correlated with liver fat content after calving in untreated cows. Our results document that sodium borate decreases the degree of fatty liver in dairy cows during early lactation.     

Fatty liver in dairy cows

An increase in liver fat concentration during the peripartum period is extremely common in dairy cows and, to some degree, is probably normal. When severe, it is associated with clinical problems including increased morbidity and mortality and reduced breeding efficiency. Fatty liver develops when serum NEFA concentrations rise and hepatic uptake of NEFA exceeds the liver's ability to synthesize and secrete lipoproteins. In most cases, hepatic lipid accumulation appears to commence prepartum, in association with rising serum NEFA concentrations and declining serum lipoprotein concentrations. Commonly used clinicopathologic tests of liver function do not yield clearly abnormal results except in animals with extremely high concentrations of liver fat. Clinically useful estimates of hepatic lipid concentration can be obtained in the field by determining the buoyancy of needle biopsy samples in liquids of various specific gravities. Clinically ill animals with liver fat concentrations of greater than 35 per cent by weight have a poor prognosis, and those that do survive will have a protracted convalescence. Treatment of dairy cows with clinical fatty liver should be aimed at reducing further adipose lipid mobilization and promoting hepatic lipoprotein synthesis; however, protocols for therapy have not as yet been evaluated critically. Prevention of fatty liver is more rewarding than treatment. Dry cows should be maintained in a moderately fat condition and fed high-quality, palatable feeds in amounts necessary to met or slightly exceed their energy requirements. Free-choice feeding of high-energy feeds should be avoided in late lactation and during the nonlactating period.     

Relevance of apolipoproteins in the development of fatty liver and fatty liver-related peripartum diseases in dairy cows

Most metabolic diseases in dairy cows occur during the peripartum period and are suggested to be derived from fatty liver initially developed during the nonlactating stage. Fatty liver is induced by hepatic uptake of nonesterified fatty acids that are released in excess by adipose tissues attributable to negative energy balance. The fatty accumulation leads to impairment of lipoprotein metabolism in the liver, and the impairment in turn influences other metabolic pathways in extrahepatic tissues such as the steroid hormone production by the corpus luteum. Detailed understanding of the impaired lipoprotein metabolism is crucial for elucidation of the mechanistic bases of the development of fatty liver and fatty liver-related peripartum diseases. This review summarizes results on evaluation of lipoprotein lipid and protein concentrations and enzyme activity in cows with fatty liver and those with ketosis, left displacement of the abomasum, milk fever, downer syndrome and retained placenta. Obtained data strongly suggest that decreases in serum concentrations of apolipoprotein B-100, apolipoprotein A-I and apolipoprotein C-III, a reduction in activity of lecithin:cholesterol acyltransferase and induction of haptoglobin and serum amyloid A are intimately related to the development of fatty liver and fatty liver-related diseases. Moreover, determination of the apolipoprotein concentrations and enzyme activity during the peripartum period is useful for early diagnoses of these diseases.     

Protein expression in dairy cows with and without subclinical hypocalcaemia

AIM: To determine differences in plasma proteomic profiles between healthy cows and those with subclinical hypocalcaemia within 12 hours after calving, and thereby explore the underlying biological mechanism of subclinical hypocalcaemia in dairy cows.

METHODS: Plasma samples were collected within 6 hours of calving from Holstein cows on a farm in Heilongjiang, China; 32 with subclinical hypocalcaemia (plasma calcium concentration 1.38–2.00?mmol/L and no clinical signs) and 59 control cows (plasma calcium concentration 2.10–2.8?mmol/L). Plasma samples were applied to weak cationic exchange protein chips for protein profiling by surface-enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF-MS), and the data were analysed using the PBS-IIC system. The amplitude of peaks for the two groups were compared using the Wilcoxon sum-rank test, and the mass-to-charge ratio of the peaks that differed was used to identify peptide fragments using the Swiss-Prot protein database.

RESULTS: Seven peaks were identified in the subclinical hypocalcaemia group that differed from those of the control group (p<0.001), that represented six unique proteins. Expression of serum albumin, fibrinogen alpha chain, amyloid beta A4 proteins and neurosecretory protein VGF were increased, and expression of apolipoprotein A-II and serum amyloid A proteins were decreased in the subclinical hypocalcaemic cows compared with control cows.

CONCLUSION: Use of SELDI-TOF-MS technology can effectively identify differences in plasma protein expression patterns in cows with subclinical hypocalcaemia. Neurosecretory protein VGF and amyloid beta A4 protein might represent useful biomarkers for diagnosis of subclinical hypocalcaemia.     

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

Samples 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.     

Macromineral status of dairy cows with concurrent left abomasal displacement and fatty liver

Abstract

AIM: To evaluate the macromineral status of field cases of dairy cows surgically treated for left abomasal displacement (LDA), with concurrent fatty liver of different severity, and compare this for animals that died or recovered.

METHODS: Sixty-eight Holstein dairy cows with LDA and 110 control cows, from 28 farms, were used in the study. Blood samples and liver biopsies were obtained during standing surgery for correction of LDA, and from control cows. The concentration of macrominerals in serum, and of total lipids (tLPD) and triglycerides (TG) in liver were determined. Liver was examined histologically, and classified for its severity of fatty liver. Cows with LDA were grouped according to severity of fatty liver. Cows in Groups 1 to 3 recovered, whereas those in Group 4 died within 4 weeks of surgery. Group 1=mild (n=4) or moderate (n=6 cows, n=4 heifers) fatty liver, Group 2 = moderate to severe fatty liver (n=13), Group 3=severe fatty liver (n=15 cows, n=5 heifers), Group 4 = severe fatty liver (n=17 cows, n=4 heifers).

RESULTS: The concentration of macrominerals in serum was affected by the concurrence of fatty liver and LDA; Ca, K and Mg were significantly (p<0.05) lower in animals that died than those that survived. For cows with severe fatty liver, concentrations of tLD and TG were higher in the animals that died compared with those that recovered (p<0.01). Cows with LDA and severe fatty liver that died were earlier in lactation (median days in milk (DIM) 13 days) compared with the other cows with LDA (median DIM 21–26 days) (p<0.05); they were also significantly older (median 6 years old) than cows in the other groups (median 4 or 5 years old) (p–0.05).

CONCLUSIONS AND CLINICAL RELEVANCE: Concentrations of macrominerals in serum were influenced by the concurrence of LDA and fatty liver. Animals with low concentrations of Ca, K and Mg had a guarded prognosis. The concentration of K should always be evaluated in cows with LDA and concurrent fatty liver when providing a prognosis. Most cows with severe fatty liver were detected in the first 4 weeks of lactation, but older animals and those that had more recently calved had a worse prognosis.     

Dry cow management for the prevention of ketosis and fatty liver in dairy cows.

Dramatic increases in energy requirements during late gestation and early lactation, superimposed on an animal with a profound drop in DMI just before calving, make the dairy cow highly susceptible to the metabolic diseases ketosis and hepatic lipidosis. Increased serum concentrations of NEFA appear to be causally linked to these problems, and feeding strategies to reduce or avoid this dramatic increase are desirable for optimal health and performance. During the last 3 to 4 weeks prepartum, a diet higher in energy and protein concentration than current NRC recommendations should be fed so that adequate nutrient intake occurs within the limits of the reduced DMI. The additional energy should be provided by glucose precursors, such as starchy concentrates or propylene glycol, and not by lipid. Excessive energy and reduced fiber should be avoided both early in the dry period (more than 28 days prepartum) and immediately postpartum. Attention should be paid to the environment of the cow, especially during the last 3 weeks prepartum, to avoid environmental stressors as much as possible.     

Use of a hand-held meter for detecting subclinical ketosis in dairy cows

The Optium Xceed is a new hand-held meter for determining blood β-hydroxybutyrate (BHBA) and glucose in human medicine. The objective of this study was to compare BHBA and glucose results obtained using the hand-held meter with those results made with a laboratory method and to evaluate its usefulness as a cowside test in the diagnosis of subclinical ketosis (SCK) in dairy cows. Seventy-eight blood samples from clinically healthy Holstein cows between 5 and 60 days post-calving were analysed. BHBA and glucose values were significantly higher with the hand-held meter versus laboratory methods. Correlation coefficients (r) for BHBA and glucose with the Optium Xceed versus laboratory methods were 0.97 and 0.63, respectively. Based on Bland–Altman plot and Passing–Bablok regression, agreement between two methods was good for BHBA but the agreement for glucose was only fair. When SCK was defined as plasma BHBA levels 1200 μmol/L, the sensitivity and specificity of the hand-held meter ketone testing in determining SCK were 85% and 94%, respectively. Raising the threshold of the laboratory method to 1400 μmol/L, the sensitivity and specificity incremented to 0.90 and 0.98, respectively.In conclusion, the blood ketone-monitoring device can be used as a rapid and reliable diagnostic test to detect SCK under field conditions.