Effects of cold treatment and ketosis induced by starvation on interferon production in leukocytes of lactating cows.

The production of interferons in blood and milk leukocytes of three groups of cows was measured to determine the effect of 6-days cold treatment (-2 degrees to -8 degrees C) and/or starving. The first group (cold) was treated with low ambient temperature (-2 degrees C to -8 degrees C) 11 hours every day for 6 days, the second (cold and starved) was treated with low temperature and starved for 6 days. The third group (controls) was fed normally and kept in a barn at room temperature (18 degrees to 20 degrees C). The leukocytes of the control and the cold treated cows responded normally to interferon induction with Newcastle Disease Virus (NDV) and mitogens: phytohemagglutinin (PHA) and concanavalin A (ConA). The cows treated with low temperature and starved for 6 days developed biochemical blood changes of ketosis. Leukocytes of these cows with ketosis produced less interferon (p less than 0.05) than before starvation and less than leukocytes of the control cows and the cold treated cows. It can be assumed that ketosis caused by starving decreases the ability of a cow's leukocytes to produce interferons.     

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%。     

Beta-hydroxybutyrate levels in peripheral blood and ketone bodies supplemented in culture media affect the in vitro chemotaxis of bovine leukocytes.

The role of ketone bodies on chemotactic capacities of leukocytes was characterized in two experiments. Experiment I was performed to investigate the association between serum beta-hydroxybutyrate concentrations (BHB) and in vitro chemotaxis of leukocytes. Cows were divided into low-BHB, medium-BHB, and high-BHB ones and classified according to their BHB. Leukocytes from high-BHB cows had a significantly lower chemotactic differential than leukocytes from low-BHB cows (p < 0.01). The effect of adding ketone bodies into in vitro chemotaxis cultures on leukocytes chemotaxis was studied in Experiment II. Either individual or a combination of commercial ketone bodies - sodium salts of BHB (BHBA), lithium salt of acetoacetate (ACAC), and acetone (Acetone) - were diluted in culture media and divided into eight concentrations corresponding to concentrations of bovine subclinical and clinical ketosis. For leukocytes from medium- and high-BHB cow, the chemotactic indexes of leukocytes were reduced by ACAC and Acetone. Chemotactic differentials of cultures with ACAC and acetone supplementation from both sources of leukocytes were significantly lower than that of the control culture (p < 0.05). For leukocytes from high-BHB cows, chemotactic indexes were suppressed in a ketone-body environment. In conclusion, leukocytes from naturally-occurring ketotic cows have lower chemotactic differentials than those from non-ketotic cows, and a chemotactic capacity indicated by a chemotactic differential is impaired when leukocytes migrate in an environment with ketone bodies in vitro.     

Subclinical ketosis in dairy cows

Subclinical ketosis is defined as a preclinical stage of ketosis. The peak prevalence of subclinical ketosis occurs during the fourth week of lactation. Herd-related factors, breed, parity, and season are other important determinants. Subclinical ketosis can be revealed by determining levels of plasma glucose, plasma NEFA and blood, and milk or urine ketone body concentration. There are theoretical and practical advantages of using milk ketone bodies. Most authors are agreed on approximate lower and upper borderlines for subclinical ketosis. The risk of an outbreak of clinical symptoms has been evaluated by some authors. Most authors have found significant negative relationships between energy balance and ketone body concentration. Some disagreement may be attributable to the fact that the diets used in different experiments can have different glucogenic potential, even if the energy content is the same. This affects the relationship between energy balance and ketone body concentration, as the ketone body level is influenced by both the energy balance and plasma glucose. Feeding silage with high butyric acid content increases the risk of subclinical ketosis. There are indications that cows with the highest milk yield directly after calving are at greatest risk for developing ketosis. Increased ketone body level secondarily reduces milk production, a decrease that has been quantified by some authors. Subclinical ketosis causes delayed reproductive functions return to normal after calving, increased intervals from calving to first and last service, and an increased frequency of ovarian cysts. The routine determination of milk acetone levels in control programs can be used to evaluate the status of individual cows, to indicate the energy feeding in early lactation at a herd level, and to evaluate sires for breeding. The heritability and the tendency toward a positive genetic correlation between milk acetone and milk yield have also been discussed, as have aspects of nutritional prevention. Factors such as energy- and protein-rich roughage, tasty high-energy concentrates, suitable feeding during the dry period, and division of the concentrates into at least four meals are considered to be important.     

Roles for Insulin and Glucagon in the Development of Ruminant Ketosis — A Review

Ketonemia can be a physiological response to a reduction in dietary intake. It also may occur when energy demands exceed the energy intake. Normally, alimentary ketogenesis is the major source of ketone bodies in ruminants. During ketonemia there is increased hepatic ketone body production. During physiological ketosis, the mobilization of free fatty acids is inadequate to support a high rate of hepatic ketogenesis. However, during clinical ketosis, the hormonal status (low insulin, high glucagon/insulin ratio) in combination with hypoglycemia promotes excessive lipid mobilization and a greater hepatic removal of fatty acids and switches the liver to a higher rate of ketogenesis. The low insulin, furthermore, can impair maximal ketone body utilization, thus exacerbating the hyperketonemia.     

Subclinical ketosis: prevalence and associations with production and disease.

Cows in 32 southern Ontario Holstein herds were monitored for subclinical ketosis for a period of two and one half years. Milk samples were routinely collected and the level of milk ketone bodies determined by the use of a commercial nitroprusside based test powder (reactions scored as negative, +1 or +2). Approximately 92% of positive reactions were observed in the first 65 days of lactation and for that time period the prevalence of ketosis was 12.1%. Based on this prevalence, the minimum possible duration of subclinical ketosis would be 7.9 days and the minimum possible lactational incidence rate would be 12.1%. The prevalence during the first 65 days of lactation in individual herds ranged from 0 to 33.9%. Subclinical ketosis was more likely to be found in cows experiencing metritis than in unaffected cows. Detection of elevated levels of milk ketones also indicated that the cow had a significantly higher risk of having clinical ketosis, metritis or cystic ovaries diagnosed within the following four days. Milk ketone scores of +1 and +2 were found to be associated with a reduction in daily milk production of 1.0 and 1.4 kg of milk respectively.     

The effect of subclinical ketosis in dry cows on the composition of the colostrum and on health indicators in newborn calves

Subclinical ketosis of dry cows was studied as to its effect on the composition of colostrum and on the health condition and selected clinico-biochemical parameters in the new-born calves of these cows. The experimental group consisted of eight cows with ketonuria and their calves. The control group consisted of six cows without ketonuria and their new-born calves. Calves of both groups were reared in similar conditions. Colostrum samples were taken from the first milking and then from the milking 24 hours after parturition. The blood of the calves was sampled before the intake of colostrum and 24 hours after birth. The concentration of ketone bodies, particularly beta-hydroxybutyrate, was much higher in the colostrum of both groups of cows than in the blood (62.6 mg total ketone bodies per 1 litre of the first colostrum of the cows of the experimental group). The concentration of ketone bodies slightly increased after parturition. The cows with ketonuria secreted into colostrum a larger amount of oxidated ketone bodies. Colostrum quality was good in both groups. Decreased viability was not recorded during the clinical examination of the calves after birth. The level of total ketone bodies in the blood of pre-colostral calves in the experimental group was 10.4 mg per litre (a much lower concentration than in blood of their dams), and exhibited no appreciable change after taking in the colostrum. The blood of calves born to cows with ketonuria contained significantly higher amounts of oxidated ketone bodies. No correlation was demonstrated between the contents of ketone bodies in the blood of cows, in the blood of calves, and in the colostrum. The calves of the experimental group were found to have a significantly higher AST activity in plasma, as compared with the control group. Twenty-four hours after birth, the quantity of immunoglobulins in the blood serum was significantly lower in the calves of the experimental group. Thanks to good attendance and hygiene, the morbidity did not increase and no losses occurred.     

不同方法检测奶牛亚临床酮病的比较

本文以试剂盒测血清β-羟丁酸含量大于1.2mmol/L作为亚临床酮病奶牛的诊断标准,采用酮粉法、试剂法、ROSS法和试纸法分别对血酮、乳酮和尿酮含量进行检测,并将定性检测的结果与定量检测的结果相比较。结果表明,酮粉法测血酮的符合率最高、为88.54%;试纸法测尿酮次之、为87.50%;ROSS法测尿酮的符合率和阳性率最低,分别为79.17%和13.54%。酮粉法测血酮敏感性最高,为70.37%;ROSS法测尿酮的敏感性最低,为37.04%;试剂法测乳酮、ROSS法测尿酮和试纸法测尿酮三者的特异性均为95.65%,高于酮粉法测血酮的特异性(94.20%),酮粉法测尿酮特异性最低、仅为88.41%。从操作难易度和成本上考虑,酮粉法测定乳酮含量比较适合牧场亚临床酮病的检测。     

Stability of plasma metabolites and hormones in parturient dairy cows

Metabolic changes that accompany the transition from parturition to lactation in dairy cows were studied. To measure these changes, plasma samples were obtained from 20 mature Holstein-Friesian dairy cows 10 days before through 10 days after parturition. They were analyzed for glucose, free fatty acids (FFA), lactic acid, ketone bodies, glucocorticoids, insulin, and growth hormone concentration. Lactic acid and glucocorticoids remained constant during the experiment, except for the day of parturition itself. In the prepartum period, changes were not detected in concentrations of hormones (glucocorticoids, insulin, and growth hormone), whereas, plasma metabolites began changing prior to parturition. Most evident were prepartum increased in FFA, ketones, and glucose. Postpartum plasma glucose concentration rapidly returned to prepartum concentrations. Plasma concentration of FFA and ketone bodies remained elevated for longer periods.     

Clinical evaluation of ketone bodies in body fluids of cows in late pregancy and their fetuses

Ketone body concentrations were determined in six body fluids of cows in late pregnancy and their fetuses (blood plasma and urine of cows, amniotic and allantoic fetal fluids, and fetal blood serum and urine) using the head-space gas chromatographic method. Physiological ratios 0/T% of oxidized (acetone and acetoacetic acid) to total (acetone, isopropanol, acetoacetic and 3-hydroxybutyric acids) ketone bodies were below 10% in cow blood plasma and urine, fetal serum and urine, and allantoic fluid, and below 20% in amniotic fluid. A scheme for the evaluation of the degree and duration of ketosis based on the 0/T% ratio is proposed. The existence of a renal regulatory mechanism for excretion of ketone bodies is suggested by analogy with the renal acidobasic regulation.     

一例奶牛酮病的诊断与防治

奶牛酮血病简称酮病,是由于奶牛体内碳水化合物及挥发性脂肪酸代谢紊乱所引起的一种全身性功能失调的代谢性疾病,本文结合病例,介绍了奶牛酮病的发病原因和临床症状,给出了几种诊疗方法。     

Effect of fasting on plasma metabolites and hormones in lactating dairy cows

Metabolic and pathophysiologic changes of secondary ketosis were studied. Plasma samples were obtained from a group of 8 mature, lactating dairy cows before, during, and after a 48-hour fast. These samples were analyzed for glucose, free fatty acids (FFA), lactic acid, ketone bodies, glucocorticoids, insulin, and growth hormone concentrations. In the prefasting period, metabolites and hormones remained constant. Lactic acid and glucocorticoids also remained stable during the entire experiment, except on the 1st day after termination of the fast, when glucocorticoids increased significantly (P less than 0.05). Glucose values decreased significantly (P less than 0.01) with the onset of fasting, began to rise halfway through the fast, and were elevated following termination of the fast. Plasma glucose concentration returned to normal by the end of the experiment. Plasma FFA concentrations increased during the early portions of the fast and decreased thereafter. Plasma ketone body concentration responded similarly, but the change occurred approximately 12 hours after the plasma FFA changes. A value determined as ketotic was reached during the fast (14.6 +/- 1.3 mg/dl). Insulin concentrations remained low during the fast and increased after fasting. Plasma growth hormone concentration increased to a new plateau in response to fasting and then decreased somewhate after fasting was ended.     

乳牛产后血浆瘦素动态与亚临床酮病的相关分析

对某奶牛场16头乳牛产前10d至产后56d血样的瘦素、酮体、葡萄糖、脂肪水平及其动态特征和相关性进行了检测。结果表明，血浆瘦素、血酮、血脂含量在不同乳牛和不同时间点之间均有显著差异。酮体在产后14～49d出现高峰，瘦素和血脂从产后第0d到产后第56d内逐渐升高。瘦素水平与血脂水平呈极显著正相关，血酮与血糖呈极显著负相关。亚临床酮病组乳牛的瘦素和血脂水平分别极显著或显著低于血酮正常组，其瘦素、血酮和血糖在试验期内的波动频率明显减少，跨度变长，瘦素在试验期内仅出现一个两端接近0、波峰不超过1．5ng／mL的波。证实，产后乳牛瘦素、血酮、血糖含量的低频率波动变化和产后8周内血浆瘦素跨时持久的单峰动态变化与乳牛亚临床酮病的发生有关。     

Composition of Milk Fat in Normal,Ketotic and Fasting Cows

By means of gas chromatography the authors have found changes in the composition of the milk fat in ketosis and after fasting, the concentration of short-chain fatty acids falling and of long-chain rising. This is interpreted as indicating that no essential difference in this respect exists between ketotic cows and cows after a period of fasting associated with hyperketonaemia. Since clinically healthy cows without hyperketonaemia likewise had a lowered concentration of short-chain and elevated concentration of long-chain fatty acids in the milk fat if they received a low-energy diet during the first weeks after calving, the authors question whether the impaired lipogenesis reported by others in primary ketosis is not a physiological consequence of fasting and therefore lacks aetiological significance for the primary ketosis.     

Prevalence of subclinical ketosis in dairy cattle in the Southwestern Iran and detection of cutoff point for NEFA and glucose concentrations for diagnosis of subclinical ketosis

Subclinical ketosis (SCK) is simply a condition marked by increased levels of circulating ketone bodies without the presence of the clinical signs of ketosis. Subclinical ketosis can cause economic losses through decreased milk production and association with preparturient diseases. Limited information is available regarding the prevalence of SCK in dairy herds in Southwestern Iran. The objectives of this study were (i) determination of the cutoff point of nonesterified fatty acids (NEFAs) and glucose concentrations for diagnosis of SCK using receiver operating characteristic (ROC) analysis, and (ii) determination of prevalence of subclinical ketosis in apparently healthy dairy cattle in Southwestern Iran. From October to December 2009, a total of 100 clinically healthy multiparous Holstein cows (3-8 years old) were randomly selected from 16 dairy herds around Kazerun, Fars Province, Iran. The cows had two-six lactations, with body weight ranging from 500 to 650 kg. Blood samples for each cow were taken at 2, 4 and 6 weeks post parturition and 3-4h after the morning feeding. The optimal cutoff point was set, by the ROC method, to >0.26 mmol/L for NEFA, and < 2.26 mmol/L for glucose with corresponding 82.54% sensitivity and 91.89% specificity for NEFA and 44.44% sensitivity and 78.38% specificity for glucose. Cows with BHB concentrations higher than 1200 μmol/L were classified as having SCK. In 2, 4 and 6 weeks post parturition 63%, 68% and 59% of the tested cows were subclinically ketotic. Overall, 97% of tested cows (97/100) were considered subclinically ketotic in at least one sample period. Thirty percent of tested cows (30/100) suffered from subclinical ketosis in all of the 2, 4 and 6 weeks postpartum. The results suggest that, a cut-off point of 0.26 mmol/L for NEFA concentrations can be used during early lactation for diagnosis of subclinical ketosis and making management decisions for prevention and treatment. Glucose cannot be a good criterion for diagnosis of SCK and it does not appear to be useful for monitoring subclinical ketosis.     

Impact of bovine somatotropin administration beginning at day 70 of lactation on serum metabolites, milk constituents, and production in cows previously exposed to exogenous somatotropin.

Metabolic and production responses are reported for 72 cows treated with bovine somatotropin (BST) for 30 days starting at day 70 of lactation. Of these 72 cows, 48 had been exposed in the preceding lactation to long-term treatment with BST at 3 dosages and 24 (controls) had not been given BST. Approximately half of the cows in each group were parity-2 cows, the rest were older. Comparisons between groups were made separately for parity-2, and older cows. Analyses, using pretreatment values of each variable as a covariate, indicated that older cows, but not parity-2 cows, significantly (P less than 0.05) increased milk production during treatment. Parity-2 cows, however, had a significantly higher milk fat percentage than controls following treatment. Cows treated with 51.6 or 86 mg BST/d in both parity groups had significantly higher serum-free fatty acids than controls. Estimated net energy balances were significantly lower for older treated cows, but did not significantly differ from controls for parity-2 treated cows. Older cows in the 86 mg of BST/d group tended to have higher concentrations of blood glucose than did older control-group cows. Treatment with BST did not significantly increase serum ketone concentrations in any group of animals, and none of the cows developed clinical ketosis during this period. Estimated net energy balance (ENEB) during treatment was a significant (P less than 0.05) covariate for free fatty acid concentrations in older cows and for milk fat percentage in parity-2 cows. Covariate adjusted analyses, using ENEB during treatment as a covariate, indicated that lipolytic stimuli already acting may be enhanced by treatment with BST, but a negative energy balance was not a necessary precondition for free fatty acid concentrations to increase following somatotropin treatment. Similarly, milk fat percentages for parity-2 treated cows were significantly (P less than 0.05) higher during treatment than controls when ENEB during treatment was used as a covariate. Increased milk fat concentrations in parity-2 treated cows were not associated with significant increases in the ratio of C18:C4-10 milk fatty acids, indicating that increased milk fat resulted from either an increase in incorporation of C18 fatty acids into milk fat coupled with an increase in de novo mammary synthesis of C4-10 milk fatty acids or an increase in C12-16 fatty acids that may arise either from increased tissue mobilization, from diet, or from de novo mammary synthesis.     

Effects of long-term feeding of a diet supplemented with clinoptilolite to dairy cows on the incidence of ketosis, milk yield and liver function

Fifty-two clinically healthy Holstein cows were randomly assigned to one of three groups according to their age and parity. The first group (A) consisted of 17 cows that were fed a concentrate ration supplemented with 1.25 per cent clinoptilolite, the second group (B) consisted of 17 cows fed a ration supplemented with 2.5 per cent clinoptilolite, and the third group (C) consisted of 18 cows, which were fed the basal ration containing no clinoptilolite. The rations were fed from four weeks before the cows' expected parturition dates until the beginning of the next dry period. Blood samples were collected from each animal at the start of the experiment, on the day of calving and then monthly, and analysed for serum glucose, ketone bodies, liver enzymes, blood urea nitrogen (BUN) and total proteins. The milk yield of each cow was recorded monthly. The cows in group B had significantly fewer cases of clinical ketosis during the first month after calving and a higher total milk yield. Feeding the cows with clinoptilolite for a long period had no apparent adverse effects on their liver function, and did not significantly affect the concentrations of glucose, ketone bodies, BUN and total proteins in their serum.     

Relationship between ketosis and dairy cows’ blood metabolites in intensive production farms of the periurban area of Dakar

This study which involved 140 Holstein and Montbeliard was carried out in the periurban area of Dakar with the aim to establish the relationship between ketosis, milk production and biochemical blood metabolites. The results showed that ketosis is a real problem in periurban farms around the city of Dakar with high proportions of 33.57% for subclinical ketosis and 6.43% for clinical ketosis. In their second month of milking, cows with subclinical ketosis had a decrease of 12.4 and 15.,6% in milk yield respectively for Montbeliard and Holstein, whereas cows with clinical ketosis had a decrease of 18.6 and 26%. Ketogenic cows (subclinical and clinical) have significantly lower average levels of blood glucose (p<0.05) and significantly higher average levels of blood urea (p<0.05) than cows with normal blood beta-Hydroxy Butyrate (BbHB) levels. Also, from one farm to another, significant difference was recorded with concentration of total proteins and globulin, calcium and magnesium.     

Disturbed bovine mitochondrial lipid metabolism: a review

In mammals, excess energy is stored primarily as triglycerides, which are mobilized when energy demands arise and cannot be covered by feed intake. This review mainly focuses on the role of long chain fatty acids in disturbed energy metabolism of the bovine species. Long chain fatty acids regulate energy metabolism as ligands of peroxisome proliferator-activated receptors. Carnitine acts as a carrier of fatty acyl groups as long-chain acyl-CoA derivatives do not penetrate the mitochondrial inner membrane. There are two different types of disorders in lipid metabolism which can occur in cattle, namely the hypoglycaemic-hypoinsulinaemic and the hyperglycaemic-hyperinsulinaemic type with the latter not always associated with ketosis. There is general agreement that fatty acid β-oxidation capability is limited in the liver of (ketotic) cows. In accord, supplemental L-carnitine decreased liver lipid accumulation in periparturient Holstein cows. Of note, around parturition concurrent oxidation of fatty acids in skeletal muscle is highly activated. Also peroxisomal β-oxidation in liver of dairy cows may be part of the hepatic adaptations to a negative energy balance (NEB) to break down fatty acids. An elevated blood concentration of nonesterified fatty acids is one of the indicators of NEB in cattle among others like increased β-hydroxy butyrate concentration, and decreased concentrations of glucose, insulin, and insulin-like growth factor-I. Assuming that liver carnitine concentrations might limit hepatic fatty acid oxidation capacity in dairy cows, further study of the role of acyl-CoA dehydrogenases and/or riboflavin in bovine ketosis is warranted.     

Hyperketonemia and the impairment of udder defense: a review

The objective of this study was to review the possible relationships between hyperketonemia and the function of phagocytes with respect to the bovine udder defense mechanism. We hypothesize that an increased incidence of clinical mastitis in high-producing cows is caused by the impairment of the udder defense mechanism during hyperketonemia. First, we review the acute phase of udder defense mechanisms after intramammary infection. The physiological changes of cows in negative energy balance are subsequently discussed. Finally, possible relationships between udder defense and physiological changes during negative energy balance, especially hyperketonemia, are reviewed. The three stages of an acute phase of udder defense are: (1) immediately eliminating invading pathogens by phagocytes, (2) releasing inflammatory substances, especially chemoattractants, and (3) migration of polymorphonuclear leukocytes into the infected udder. Leukocytes from hyperketonemia subjects show a lower capacity of the phagocytic defense mechanism. In addition, the phagocytic and bactericidal capacities of neutrophils are reduced when these cells are acting in the presence of high concentrations of ketone bodies. Lower amounts of cytokine production after bacterial infection are observed in ketotic subjects. The chemotactic capacity of blood leukocytes is impaired in leukocytes obtained from ketotic cows. Lower numbers of blood leukocytes are observed in ketotic cows. In conclusion, the impairment of the udder defense mechanism in negative energy balance cows seems related to hyperketonemia.