R. Jorritsma

Prevalence and indicators of post partum fatty infiltration of the liver in nine commercial dairy herds in The Netherlands

Abstract A field study was performed with 218 dairy cows in nine dairy herds in order to investigate the prevalence of post partum fatty infiltration of the liver and its relationship to subsequent body condition scores, blood variables and milk production. The mean concentration of triacylglycerols in the liver was 61.2 mg/g wet liver tissue. The prevalence of fatty liver (more than 50 mg triacylglycerol in 1 g wet liver tissue) was 54.1%. Serum non-esterified fatty acids, urea and blood glucose concentrations appeared to be significant indicators of hepatic lipidosis between 6 and 17 days post partum (R2=0.33). High milk production and large losses of body condition score in early lactation were significant indicators of hepatic lipidosis from a retrospective point of view (R2=0.22). Single body condition scores were not significantly related to the concentration of triacylglycerol in the liver. It was concluded that, according to the classification used, fatty liver seems to be fairly common in early lactating dairy cows. Although correlation coefficients were too low to enable a precise and accurate description of liver triacylglycerol content, we found some biologically explicable variables in commercial herds that were significantly related to the liver triacylglycerol content.

Relationships between fatty liver and fertility and some periparturient diseases in commercial Dutch dairy herds

Declining fertility in dairy cows is frequently suggested to arise from the occurrence of a more negative energy balance and/or the concomitant increased accumulation of triacylglycerol in the liver. Therefore, we performed a field study to assess the clinical effects of postpartum fatty liver in dairy cows on fertility and reproductive disease. Data were collected from 360 cows from nine dairy herds on fertility, diseases, and the liver triacylglycerol content on two occasions during lactation: 6 to 17 days and 38 to 50 days postpartum. The mean concentration of triacylglycerol in the liver was 54.6 mg/g from 6 to 17 days and 38.4 mg/g from 38 to 50 days postpartum. The probability of pregnancy was 30% lower for cows with higher contents of triacylglycerol in the liver compared to the probability for cows with low liver triacylglycerol (P = 0.049). The probability of estrus was also 35% lower for the cows with high triacylglycerol in the liver. This resulted in larger intervals between parturition and first heat and parturition and pregnancy for these cows. There was no effect observed on the first insemination conception rate. Given a certain level of triacylglycerol, recorded milk production had a positive effect on time to pregnancy. The incidences of endometritis, lochiometra and cystic ovarian follicles were not higher in cows with higher liver triacylglycerol contents. Endometritis was associated with a lower first insemination conception rate and more days open (chi2 = 4.26, P = 0.03 and T-test = -2.02, P= 0.04 respectively). We concluded that our results support the idea that differences in the negative energy balance or the accumulation of triacylglycerol in the liver of postpartum dairy cows affect fertility performance. The data also indicate that an increase in milk production has no negative impact on fertility as long as the amount of triacylglycerol in the liver remains the same.

Prevalence, prediction and risk factors of enteropathogens in normal and non-normal faeces of young Dutch dairy calves.

Abstract Between January and April 2007, 424 calves under 22 days of age from 108 Dutch dairy herds were sampled to estimate the prevalence of non-normal faeces (‘custard-like’—yellowish-coloured with custard consistency or diarrhoea: watery-like faeces) and the shedding of enteropathogens Escherichia coli K99 (E. coli), Coronavirus, Cryptosporidium parvum (C. parvum), Rotavirus and Clostridium perfringens (Cl. perfringens). In addition, information was collected on animal characteristics and herd-management practices. The probability of detecting each one of five enteropathogens given a calf with ‘custard-like’ faeces or diarrhoea was estimated using Bayes’ rule and was based on the predicted probabilities from a multinominal model including each of five enteropathogens as independent variables. In addition, putative risk factors for the presence of each of five enteropathogens were analysed using logistic regression models with random herd effects. Fifty-seven percent of calves had faeces of normal colour (brownish) and consistency (firm), 23.8% (95%CI: 19.8–28.2%) had ‘custard-like’ faeces and 19.1% (95%CI: 15.5–23.2%) had diarrhoea. E. coli was the least detected enteropathogen (2.6% (95%CI: 1.3–4.6%) of calves, 9% (95%CI: 5–16%) of herds) and Cl. perfringens was most detected (54.0% (95%CI: 49.1–58.8%) of calves, 85% (95%CI: 77–91%) of herds). E. coli and Coronavirus were detected incidentally in only one or two calves per herd, whereas C. parvum and Cl. perfringens were frequently detected in up to four calves per herd. For calves with ‘custard-like’ faeces, the probability of detecting Rotavirus from a calf in its first week of age was 0.31 whereas for a calf in its second week, there was a 0.66 probability of detecting C. parvum. The probabilities of detecting E. coli, Rotavirus and C. parvum in calves with diarrhoea in their first week of age were 0.10, 0.20 and 0.43, respectively. In calves with diarrhoea between 1 and 2 weeks of age, the probability of detecting enteropathogens was 0.43 for C. parvum. None of the tested enteropathogens were related to ‘custard-like’ faeces or diarrhoea in the third week of age. Putative risk factors for E. coli, Coronavirus and C. parvum included the presence of peer-calves shedding Coronavirus, C. parvum or Rotavirus, respectively. Additionally, managerial risk factors such as non-optimal hygienic housing (for Coronavirus) and the routine use of antibiotics for diarrhoeic calves (for C. parvum) were found. No animal or managerial factors were associated with shedding of Cl. perfringens.

Acute fasting in heifers as a model for assessing the relationship between plasma and follicular fluid NEFA concentrations.

It is known from epidemiological studies that negative energy balance in early lactating dairy cows is related to a depression in reproductive performance. Elevated plasma concentrations of non-esterified fatty acids (NEFA) are a typical metabolic characteristic of these animals and are proposed as the possible link. The suggestion is that NEFA might have a direct effect on the ovary, by affecting development of the oocyte or the granulosa cells. However, no data is available concerning the relationship between the concentration of NEFA in follicular fluid and plasma. Therefore, a cross-over study with 10 heifers around 15 months of age was conducted to analyze this relationship and examine the suggested effects of the negative energy balance on follicular growth. Investigation of these effects was performed on fasted heifers. The experimental treatment consisted of feeding hay with a subsequent period of fasting, to induce elevated plasma NEFA concentrations. Sampling of follicular fluid was performed using transvaginal aspiration of follicles, which were standardized using a synchronization protocol. In addition, concentrations of glucose, insulin, NEFA, and estradiol were measured in plasma. Follicular estradiol and progesterone concentrations were also measured to assess the quality of the dominant follicle. Fasting resulted in significantly lower plasma glucose (P=0.0006) and plasma insulin (P<0.0001) concentrations, higher plasma estradiol (P=0.008) and higher NEFA (P<0.001) concentrations, and smaller follicles (P=0.04) with lower estradiol:progesterone (E/P) ratios (P=0.05). Concentrations of NEFA in follicular fluid and plasma were closely related. Given this close relationship, we concluded that the presence of high plasma NEFA concentrations might link energy metabolism in early lactation with fertility.

Activities of the enzymes of hepatic gluconeogenesis in periparturient dairy cows with induced fatty liver.

The objective was to measure the activities of all the enzymes essential for hepatic gluconeogenesis in dairy cows with induced fatty liver. We aimed to induce severe fatty liver in ten experimental cows by overfeeding them during the dry period while seven control cows were maintained on a restricted diet. To induce a marked negative energy balance, the experimental cows were deprived of feed for 8 h immediately after parturition. In addition, the experimental cows were given a restricted amount of diet during the first 5 d of lactation. Liver samples were collected 1 week before and 1, 2 and 4 weeks after parturition. Before parturition, liver triacylglycerol concentrations did not differ between the two groups. After parturition, the experimental cows developed marked fatty liver as indicated by a higher level of triacylglycerols in the liver compared with the control cows. Before parturition, all gluconeogenic enzymes in the liver were lower in experimental cows than in control cows. Phosphoenolpyruvate carboxykinase, pyruvate carboxylase and propionyl-CoA carboxylase were significantly lower and fructose 1,6-bisphosphatase and glucose 6-phosphatase tended to be lower in the experimental cows. The activities of two crucial enzymes for gluconeogenesis in ruminants, i.e., phosphoenolpyruvate carboxykinase and propionyl-CoA carboxylase, remained low throughout the sampling period post partum. Activities of pyruvate carboxylase and glucose 6-phosphatase in the experimental cows post partum were upgraded to values similar to those of the control cows. The results showed that the capacity for hepatic gluconeogenesis before parturition was lower in cows with induced fatty liver than in control cows. After parturition, the low activities of crucial gluconeogenic enzymes indicated insufficient production of glucose. It is suggested that the low gluconeogenic capacity leads successively to low blood glucose concentrations, low insulin levels and high rates of mobilization of fatty acid, causing severe hepatic lipidosis.

Routine detection of hyperketonemia in dairy cows using Fourier transform infrared spectroscopy analysis of β-hydroxybutyrate and acetone in milk in combination with test-day information

The objective of this study was to assess the quality of a diagnostic model for the detection of hyperketonemia in early lactation dairy cows at test days. This diagnostic model comprised acetone and β-hydroxybutyrate (BHBA) concentrations in milk, as determined by Fourier transform infrared (FTIR) spectroscopy, in addition to other available test-day information. Plasma BHBA concentration was determined at a regular test day in 1,678 cows between 5 and 60 d in milk, originating from 118 randomly selected farms in the Netherlands. The observed prevalence of hyperketonemia (defined as plasma BHBA ≥1,200 µmol/L) was 11.2%. The value of FTIR predictions of milk acetone and milk BHBA concentrations as single tests for hyperketonemia were found limited, given the relatively large number of false positive test-day results. Therefore, a multivariate logistic regression model with a random herd effect was constructed, using parity, season, milk fat-to-protein ratio, and FTIR predictions of milk acetone and milk BHBA as predictive variables. This diagnostic model had 82.4% sensitivity and 83.8% specificity at the optimal cutoff value (defined as maximum sum of sensitivity and specificity) for the detection of hyperketonemia at test days. Increasing the cutoff value of the model to obtain a specificity of 95% increased the predicted value of a positive test result to 56.5%. Confirmation of test-positive samples with wet chemistry analysis of milk acetone or milk BHBA concentrations (serial testing) improved the diagnostic performance of the test procedure. The presented model was considered not suitable for individual detection of cows with ketosis due to the length of the test-day interval and the low positive predictive values of the investigated test procedures. The diagnostic model is, in our opinion, valuable for herd-level monitoring of hyperketonemia, especially when the model is combined with wet chemistry analysis of milk acetone or milk BHBA concentrations. By using the diagnostic model in combination with wet chemistry milk BHBA analysis, 84% of herds were correctly classified at a 10% alarm-level prevalence. As misclassification of herds may particularly occur when only a limited number of fresh cows are sampled, we suggest using prevalence estimates over several consecutive test days to evaluate feeding and management practices in smaller dairy farms.

Analysis of the economically optimal voluntary waiting period for first insemination

The voluntary waiting period (VWP) is defined as the time between parturition and the time at which the cow is first eligible for insemination. Determining the optimal VWP from field data is difficult and unlikely to happen. Therefore, a Monte-Carlo dynamic-stochastic simulation model was created to calculate the economic effects of different VWP. The model is dynamic and uses time steps of 1 wk to simulate the reproductive cycle (ovulation, estrous detection, and conception), the occurrence of postpartum disorders, and the lactation curve. Inputs of the model were chosen to reflect the situation of Dutch dairy cows. In the model, we initially created a cow of a randomly selected breed, parity, month of calving, calf status of last calving, and expected 305-d milk yield. The randomly varied variables were based upon relevant distributions and adjusted for cow statuses. The lactation curve was modeled by Woods function. The economic input values in the analysis included: cost of milk production (€0.07 to €0.20 per kg), calf price (€35 to €150 per calf), AI cost (€7 to €24 per AI), calving management cost (€137 to €167 per calving), and culling cost, expressed as the retention pay-off (€118 to €1,117). A partial budget approach was used to calculate the economic effect of varying the VWP from 7 to 15 wk postpartum, using a VWP of 6 wk as reference. Per iteration, the VWP with either the lowest economic loss or the maximum profit was determined as the optimal VWP. The optimal VWP of most cows (90%) was less than 10 wk. On average, every VWP longer than 6 wk gave economic losses. Longer VWP were in particular optimal for the first parity of breeds other than Holstein-Friesian, cows calving in winter with low milk production, high milk persistency, delayed peak milk yield time, a delayed time of first ovulation, or occurrence of a postpartum disorder, and while costs of milk production are low and costs for AI are high.

Genetic and nongenetic variation in plasma and milk β-hydroxybutyrate and milk acetone concentrations of early-lactation dairy cows

This study assessed genetic variation, heritability estimates, and genetic correlations for concentrations of plasma β-hydroxybutyrate (BHBA), milk BHBA, and milk acetone in early lactation to investigate differences between cows in susceptibility to hyperketonemia and possibilities to use test-day milk ketone bodies for genetic improvement. Blood and test-day milk samples were collected on randomly selected dairy farms in the Netherlands from cows of various parities between 5 and 60 d in milk. Plasma samples were analyzed for BHBA (reference test for hyperketonemia) and test-day milk samples were analyzed for BHBA and acetone using Fourier-transform infrared spectroscopy. The final data set consisted of plasma BHBA concentrations of 1,615 cows from 122 herds. Milk BHBA and milk acetone concentrations were determined for 1,565 cows. Genetic variation, heritability, and proportion of phenotypic variation attributable to the herd were estimated using an animal model with fixed effects for parity and season, a covariate for days in milk, and random effects for herd, animal, and error. Genetic correlations for plasma BHBA, milk BHBA, and milk acetone were estimated using bivariate analyses. The heritability estimate for plasma BHBA concentrations in early lactation was 0.17, whereas heritability estimates for milk BHBA and milk acetone were 0.16 and 0.10, respectively. This indicates that selective breeding may contribute to a lower incidence of hyperketonemia in early lactation. For the 3 traits, the proportion of variance attributable to herd was larger than the additive genetic variance, underlining the importance of on-farm feeding and management in the etiology of hyperketonemia in fresh cows. Prevention strategies for hyperketonemia can, therefore, include both feeding and management strategies at dairy farms (short-term) and genetic improvement through breeding programs (long-term). Genetic correlations between concentrations of plasma BHBA and milk BHBA (0.52) or milk acetone (0.52) were moderate. As milk ketone bodies can be routinely analyzed at test days, this may provide a practical alternative for breeding programs aimed at reducing hyperketonemia in early lactation.

Evaluation of a milk test for detection of subclinical ketosis

This study included 190 lactating cows from eight dairy farms and was designed to investigate the sensitivity and specificity of a milk test (Ketolac) for detecting subclinical ketosis in dairy cattle. Blood and milk samples were collected between 38 and 50 days post partum. At a beta-hydroxybutyric acid concentration in blood of 1.2 mmol/L, the prevalence of ketosis was 14%. A Response Operator Characteristic curve was drawn for two milk test values: 100 mumol/L and 200 mumol/L. Assessment of several beta-hydroxybutyric acid cut-off values in blood showed that when 100 mumol/L on the Ketolac milk test was defined positive, the most desirable sensitivity-specificity combination was achieved. This milk test can be routinely used to screen a herd for subclinical ketosis.

Effect of milk yield characteristics, breed, and parity on success of the first insemination in Dutch dairy cows

The objective of this study was to determine the contribution of cow factors to the probability of a successful first insemination (SFI). The investigation was performed with 51,791 lactations from 1,396 herds obtained from the Dutch dairy cow database of the Cattle Improvement Co-operative (CRV). Cows that had the first insemination (AI) between 40 and 150 d postpartum were selected. The first AI was classified as successful when cows were not reinseminated and either calved between 267 and 295 d later or were culled within 135 to 295 d after first AI. The lactation curve characteristics of individual lactations were estimated by Wilminks curve using the test-day milk records from CRV. The lactation curve characteristics (peak milk yield, milk yield at the first-AI date, time of peak yield (PT), and milk persistency) were calculated. Breed, parity, interval from calving to first AI (CFI), lactation curve characteristics, milk production traits, moment of AI related to PT (before or after PT), calf status, month of AI, and month of calving were selected as independent variables for a model with SFI as a dependent variable. A multivariable logistic regression model was used with farm as a random effect. Overall SFI was 44%. The effect of parity on SFI depended on CFI. The first-parity cows had the greatest SFI (0.43) compared with other parities (0.32-0.39) at the same period of CFI before 60 d in milk (DIM), and cows in parity ≥5 had the least SFI (0.38-0.40) when AI was after 60 DIM. After 60 DIM, extending CFI did not improve SFI in the first-parity cows, but SFI was improved in multiparous cows. Holstein-Friesian cows had lesser SFI (0.37) compared with cross-breed cows (0.39-0.46). Twin and stillbirth calving reduced SFI (0.39) compared with a single female calf (0.45) or a male calf (0.43) calving. The SFI in different months of AI varied and depended on CFI. Cows that received AI before 60 DIM had a lesser SFI, especially in March, June, and July (0.18, 0.35, and 0.34, respectively). Artificial insemination before PT reduced SFI (0.39) in comparison with AI after PT (0.44). The effect of milk yield at the first-AI date on SFI varied depending on CFI. After 60 DIM at the same period of CFI, a high level of milk yield at the first-AI date reduced SFI. In conclusion, knowledge of the contribution of cow factors on SFI can be applied to support decision making on the moment of insemination of an individual cow in estrus.