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ABSTRACT: This open-label, randomized clinical trial with positive control compared the treatment of active digital dermatitis (DD) lesions (stages M1, M2, and M4.1) on dairy cattle hind feet with an enzyme alginogel or a copper and zinc chelate gel (coppergel). Upon recruitment (d 0), active DD lesions were cleaned, photographed, treated, and bandaged. This procedure was repeated on d 3 and d 7, with treatment and bandaging discontinued for those lesions that had transitioned to the M0, M3, or M4 stage on d 7. Day 10 was considered the end of the treatment trial, and all recruited feet were cleaned and photographed. Treatment effect of the 2 products was assessed not only using the M-score but also using general wound healing progress criteria. Improvement of M-score was defined as transition to M0, M3, or M4 stages, or to lesions with a smaller ulcerative area (e.g., M2 stage to M1 stage). Lesions with improved wound healing had at least one of the following criteria when compared with the previous observation: decreased defect size, healthier granulation tissue color (pink-red instead of purple-grayish), more regular aspect of granulation tissue surface, wound contraction, or epithelization starting from the surrounding skin. Both primary outcomes were assessed using a multivariable logistic regression analysis. Lesions treated with the enzyme alginogel had a decreased adjusted odds ratio for M-score improvement (aOR: 0.04; 95% confidence interval: 0.01–0.11). Lesions treated with the coppergel mostly transitioned to chronic lesions, whereas lesions treated with the enzyme alginogel mostly remained active lesions. The wound healing progress of almost 70% of the lesions treated with coppergel could not be scored, for the greater part due to the presence of crust materials. With these unscorable lesions classified as “improved,” there was no treatment effect on wound healing progress (aOR: 0.99; 95% confidence interval: 0.34–3.05), whereas with unscorable lesions classified as “not improved,” the enzyme alginogel outperformed the coppergel with regard to wound healing progress (aOR: 2.48; 95% confidence interval: 1.07–5.79). None of the products used in our study achieved high cure rates (transition to the M0 stage) for active DD lesions. Low cure rates of topical treatment of DD, together with the important role of chronic lesions in the epidemiology of DD, indicate that future research should investigate how to achieve successful wound management of DD lesions, thereby mitigating pain associated with the lesions and reducing both transmission and prevalence of DD within herds.

Dairy cows that are unable to adapt to a change in their metabolic status are at risk for hyperketonaemia (HK). Reported HK herd level prevalences range a lot and we hypothesized that this is partly due to differences in used tests and monitoring protocols. Insights in milk β-hydroxybutyrate (BHB) metrics can potentially explain why the reported incidences or prevalences vary between test strategies. Automated collection and repeated analyses of individual milk samples with the DeLaval Herd Navigator ™ (HN) provides real-time data on milk BHB concentrations. We aimed to use that information to gain insight in BHB metrics measured in milk from 3 to 60 days in milk (DIM). Using different cut-offs (0.08, 0.10 and 0.15 mmol/L), 5 BHB metrics were determined. Furthermore, the impact of 4 arbitrary test protocols on the detected incidence of HK was assessed. We used HN data of 3,133 cows from 35 herds. The cumulative incidence of HK between 3 and 60 DIM varied between 30.5 and 76.7% for different cut-off values. We found a higher HK incidence for higher parity cows. The first elevated BHB concentrations were roughly found between one and two weeks after calving. For higher parity cows the maximum BHB concentrations were higher, the onset of HK was earlier after calving, and the number of episodes of HK was higher. It appeared that the sensitivity of a HK test protocol can be increased by increasing the testing frequency from once to twice a week. Also extending the number of days of the test window from 4-14 to 4-21 days enhances the chance to find cows experiencing HK. In conclusion, HN data provided useful insights in milk BHB metrics. The chosen cut-off value had a large effect on the reported metrics which explains why earlier reported incidences or prevalences vary such a lot. Differences in test period and sample selection also had a large impact on the observed HK incidence. We suggest to take this in consideration while evaluating whether HK is an issue on farm level and use a uniform protocol for benchmarking of HK between farms., Competing Interests: AR-D is employee at Elanco Benelux and HV is employee at DeLaval Benelux. EJ was student when she performed the study. WS and RJ supervised EJ and received no additional funding. The authors declare that this study received funding from Delaval Benelux and Elanco Benelux. The funder had the following involvement in the study: article processing costs., (Copyright © 2023 De Jong, Rijpert-Duvivier, Veldman, Steeneveld and Jorritsma.)

Introduction: While prevention is increasingly important in the dairy sector, implementation of cost-effective preventive measures is often lacking. To increase the use of these measures and consequently improve animal welfare and reduce financial losses for farmers, it is necessary to know the drivers and constraints of farmers to engage in prevention., Methods: Therefore, we invited farmers to participate in an online questionnaire, which contained questions about their behavior toward either claw health or calf health. We used the theory form the Stage of Change model, COM-B, as well as the Theory of Planned Behavior to formulate our questions. We used the responses of 226 farmers in our analyses, who were equally distributed over the two groups of diseases., Results and Discussion: We found that 63.5% of responding farmers were in the action phase or the maintenance phase to prevent claw diseases and even more (85.4%) to prevent calf diseases. The responses also suggest that many farmers have the knowledge and skills to implement preventive measures for both claw and calf diseases. The scores for social and physical opportunities for calf diseases were significantly higher than for claw diseases and all other COM-B components were also numerically higher for calf diseases. This suggests that farmers' perception of taking preventive measures against claw diseases is more difficult than taking preventive measures against calf disease. The automation of preventive behavior scored relatively low for both groups of diseases, which suggests that farmers may need reminders to persist in their activities and support to create habitual prevention behaviors. From these results, we concluded that creating social norms, supporting discussions among farmers, and using environmental adaptations may result in more preventive behavior., Competing Interests: JS was employed by MSD Animal Health. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Jorritsma, Swinkels, Werven, Lahaye, Martena and Stok.)

An extended calving interval (CInt) by extending the voluntary waiting period (VWP) could be associated with altered metabolism in dairy cows. The aim of this study was first to evaluate the effects of VWP on metabolism and body condition during the first 305 d after the first calving in the experiment (calving 1), around the end of the VWP, and during pregnancy (280 d before calving 2). Second, the effects of the VWP on metabolism were determined from 2 wk before until 6 wk after calving 2. Third, individual cow characteristics were used to predict milk production and body condition of cows after different VWP. Holstein-Friesian cows (N = 154, 41 primiparous [PP], 113 multiparous [MP]) were blocked for parity, milk production, and lactation persistency, randomly assigned to a VWP of 50, 125, or 200 d (VWP50, VWP125, or VWP200) and followed from calving 1 until 6 wk after calving 2. In the first 6 wk after calving 1 and from 2 wk before until 6 wk after calving 2, weekly plasma samples were analyzed for nonesterified fatty acids (NEFA), β-hydroxybutyrate, glucose, insulin, and insulin-like growth factor 1 (IGF-1). From wk 7 after calving 1 until 2 wk before calving 2, insulin and IGF-1 were analyzed every 2 wk. Fat- and protein-corrected milk (FPCM) and body weight (BW) gain were measured weekly. Cows were divided in two parity classes based on calving 1 (PP and MP) and remained in these classes after calving 2. During pregnancy, MP cows in VWP200 had greater plasma insulin and IGF-1 concentration and lower FPCM compared with MP cows in VWP125 (insulin: 18.5 vs. 13.9 µU/mL, CI 13.0-19.7, P < 0.01; IGF-1: 198.5 vs. 175.3 ng/mL ± 5.3, P = 0.04; FPCM: 22.6 vs. 30.0 kg/d ± 0.8, P < 0.01) or VWP50 (insulin: 15.8 µU/mL, P < 0.01; IGF-1: 178.2 ng/mL, P < 0.01; FPCM: 26.6 kg/d, P < 0.01) and had a greater daily BW gain compared with cows in VWP50 (3.6 vs. 2.5 kg/d ± 0.2; P < 0.01). After calving 2, MP cows in VWP200 had greater plasma NEFA concentration (0.41 mmol/liter) compared with MP cows in VWP125 (0.30 mmol/liter, P = 0.04) or VWP50 (0.26 mmol/liter, P < 0.01). For PP cows, the VWP did not affect FPCM or body condition during the first lactation in the experiment, or metabolism after calving 2. Independent of the VWP, higher milk production and lower body condition before insemination were associated with higher milk production and lower body condition at the end of the lactation. Variation in these characteristics among cows could call for an individual approach for an extended VWP., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Colostrum intake is one of the most important factors in neonatal health in ruminants, mainly because of its unique immunological properties. Both in practice as well as in research, the attention of lactogenic immunity is focused on the importance of colostral antibodies and less attention is given to the functional role of maternal cells in colostrum. Here we study the transfer of maternal leukocytes via colostrum and the functionality in goat kids. In experiment 1, twenty twin pairs of goat kids from dams previously immunized with an inactivated Mycobacterium avium subsp. paratuberculosis (MAP) vaccine were fed maternal colostrum from their dam (kid 1) or pasteurized and frozen/thawed bovine colostrum (kid 2). The presence of cell mediated immune response (CMIR) against Mycobacterium avium antigens in the kids was assessed using intradermal skin testing with PPD-A tuberculin. Linear mixed effect models showed an increase in skin thickness in response to intradermal PPD-A injection in maternal colostrum fed kids compared to bovine colostrum fed kids. After intradermal PPD-A application, serum concentration of MAP specific antibodies increased in kids fed maternal colostrum, indicating antigen specific activation of the adaptive immune system. We did not detect a similar increase in antibodies in the kids fed bovine colostrum. In experiment 2, a more reductionistic approach was applied to specifically study the effects of the transfer of maternal colostral leukocytes on CMIR in goat kids. Similar to experiment 1, twin kids from MAP immunized dams were randomly divided over two groups. The experimental group received colostrum replacer supplemented with fluorescently labelled colostral cells of the dam and the control group received colostrum replacer only. No difference in skin response following intradermal PPD-A injection was observed between both groups of kids. Histologic examination of the skin at the intradermal injection site did not show fluorescently labelled cells. In conclusion, in our initial experiment we observed an antigen specific CMIR in goat kids fed fresh colostrum with colostral leukocytes from vaccinated dams. The lack of a DTH response in kids fed colostrum replacer supplemented with maternal colostrum derived leukocytes indicated that the complete colostral matrix is probably required for colostrum leukocytes to transfer across the intestinal epithelial barrier and modulate the neonatal immune response. In line with earlier studies, our results indicate that caprine maternal leukocytes present in colostrum can functionally contribute to the newborns' early adaptive immune responses adding to the importance of colostrum feeding in ruminant neonates., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

This open-label, randomized clinical trial with positive control compared the treatment of active digital dermatitis (DD) lesions (stages M1, M2, and M4.1) on dairy cattle hind feet with an enzyme alginogel or a copper and zinc chelate gel (coppergel). Upon recruitment (d 0), active DD lesions were cleaned, photographed, treated, and bandaged. This procedure was repeated on d 3 and d 7, with treatment and bandaging discontinued for those lesions that had transitioned to the M0, M3, or M4 stage on d 7. Day 10 was considered the end of the treatment trial, and all recruited feet were cleaned and photographed. Treatment effect of the 2 products was assessed not only using the M-score but also using general wound healing progress criteria. Improvement of M-score was defined as transition to M0, M3, or M4 stages, or to lesions with a smaller ulcerative area (e.g., M2 stage to M1 stage). Lesions with improved wound healing had at least one of the following criteria when compared with the previous observation: decreased defect size, healthier granulation tissue color (pink-red instead of purple-grayish), more regular aspect of granulation tissue surface, wound contraction, or epithelization starting from the surrounding skin. Both primary outcomes were assessed using a multivariable logistic regression analysis. Lesions treated with the enzyme alginogel had a decreased adjusted odds ratio for M-score improvement (aOR: 0.04; 95% confidence interval: 0.01-0.11). Lesions treated with the coppergel mostly transitioned to chronic lesions, whereas lesions treated with the enzyme alginogel mostly remained active lesions. The wound healing progress of almost 70% of the lesions treated with coppergel could not be scored, for the greater part due to the presence of crust materials. With these unscorable lesions classified as "improved," there was no treatment effect on wound healing progress (aOR: 0.99; 95% confidence interval: 0.34-3.05), whereas with unscorable lesions classified as "not improved," the enzyme alginogel outperformed the coppergel with regard to wound healing progress (aOR: 2.48; 95% confidence interval: 1.07-5.79). None of the products used in our study achieved high cure rates (transition to the M0 stage) for active DD lesions. Low cure rates of topical treatment of DD, together with the important role of chronic lesions in the epidemiology of DD, indicate that future research should investigate how to achieve successful wound management of DD lesions, thereby mitigating pain associated with the lesions and reducing both transmission and prevalence of DD within herds., (© 2022, The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This research paper addresses the hypothesis that cow introductions in dairy herds affect milk production and behaviour of animals already in the herd. In dairy farms, cows are commonly regrouped or moved. Negative effects of regroupings on the introduced animals are reported in other studies. However, little is known about the effects on lactating cows in the herd. In this research a herd of 53 lactating dairy cows was divided into two groups in a cross-over design study. 25 cows were selected as focal cows for which continuous sensor data were collected. The treatment period consisted of replacing non-focal cows three times a week. Many potentially influencing factors were taken into account in the analysis. Replacement of cows in the treatment period indeed affected the focal animals. During the treatment period these cows showed increased walking and reduced rumination activity and produced less milk compared to the control period. Milk production per milking decreased in the treatment period up to 0.4 kg per milking on certain weekdays. Lying and standing behaviour were similar between the control and the treatment period. The current study suggests that cow introductions affect welfare and milk production of the cows already in the herd.

There is increased concern about the selection pressure of antimicrobial use (AMU) in humans as well in farm animals resulting in antimicrobial (AM) resistance. The introduction of monitoring of AMU in food-producing animals since 2011 has led to a considerable quantitative reduction of AMU in those animal species in the Netherlands. This survey was conducted to explore the possibilities to improve prudent use of AM in the cattle industry. We sent an online questionnaire to 373 veterinarians and asked which antimicrobial prophylaxis they used for their most recently performed cesarean section (CS) and left displaced abomasum (LDA) correction. With a response rate of 30%, we found that older graduates used more AM for CS than recently graduated veterinarians (odds ratio = 2.4 to 2.7 depending on category), whereas antimicrobial prophylaxis in LDA surgeries was significantly different for the available surgical correction methods. The results indicate that the respondents do not adjust the surgical antimicrobial prophylaxis for different conditions of the patient or the environment; 37 (38%) respondents consider that the risks of contamination are similar for CS and LDA. In CS and all LDA correction methods preoperative prophylaxis was significantly less often applied than postoperative antimicrobial treatments (odds ratio = 0.05 and 0.08, respectively). When preoperative prophylaxis was applied, the choice of (combinations of) the AM and the timing of administration were not adequate at the start of the surgical procedure (88% for CS and 90% for LDA). We conclude that considerable qualitative (timing, choice of antimicrobial, route of administration) and quantitative (limit AMU to indicated procedures) improvement on antimicrobial prophylaxis for CS and LDA is possible by appreciation of the risks of the type of surgery (CS, LDA) and its conditions as well as by selecting classes of AM and timing and routes of administration that result in effective drug concentrations at the start of the procedure., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).)

Newborn calves are agammaglobulinemic and rely for their first immune protection almost completely on the transfer of immune constituents via colostrum. Inadequate colostrum management practices such as on-farm colostrum storage practices and colostrum feeding methods could affect immune components in colostrum and subsequently immune status of the newborn calf. We conducted a scoping review to identify all literature on the interactions between several colostrum management factors and immunological colostrum quality and passive transfer of immunity. Three major stages were defined: milking methods, colostrum treatment and storage, and administration procedures. Separate CAB Abstracts searches were performed for each of the subjects of interest. The search process was completed on November 9, 2020. Colostrum should be milked as soon as possible, as IgG concentration diminishes over time, probably due to dilution. To minimize bacterial contamination, it is advised to pasteurize colostrum in small batches at maximal 60°C for 30 or 60 min. Freeze/thawing of colostrum does not or only slightly affect IgG concentrations, as long as thawing is done au bain-marie and temperature does not exceed 40°C. In on-farm situations, it is difficult to determine the volume that should be fed as the variables contributing to the absorption of IgG by the newborn calf are many and include the quality of the colostrum, the bacterial contamination, the time interval between birth and first moment of feeding and the weight of the calf. Despite all knowledge regarding optimal colostrum management strategies, it remains challenging to predict the effects of certain colostrum management choices in field conditions. Therefore, we recommend measuring the colostral quality, weighing the newborn calf, adjusting the feeding volume accordingly to ensure optimal colostrum intake for each calf., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Robbers, Jorritsma, Nielen and Koets.)

Colostrum feeding is essential for the transfer of passive immunity and health of newborn calves. Information on current colostrum management practices to reduce calf morbidity and mortality is important but lacking for Dutch dairy herds. We therefore conducted a survey to investigate colostrum management strategies on Dutch dairy farms. The survey was specifically focused on the most recently born calf and was returned by 107 respondents (response rate of 13.4%). The mean amount of colostrum fed at first feeding was 2.9 liters. Overall, 79% of farmers provided the calf with at least 6 liters of colostrum in up to three feedings. The majority of respondents (84%) claimed to provide the calf with colostrum for the first time within 2 h post-partum. Using ordinal logistic regression and Wilcoxon rank sum test, we found no differences in time to first colostrum feeding or total amount of colostrum fed between bull calves and heifer calves, respectively. Ordinal logistic regression showed no significant differences in time to first colostrum feeding or time between calving and removing the calf from the dam between AMS and conventional milking herds. Two sample T -test comparing the total volume of colostrum showed no significant difference between AMS and conventional milking herds. Time of day at which a calf was born affected both volume fed at first colostrum feeding and time until first colostrum feeding. Calves born between 00.00 and 06.00 were significantly at risk of receiving the first colostrum later as compared to calves born at other times. Calves born in the evening received on average a lower amount of colostrum at first feeding. Survey results on colostrum management on most Dutch dairy farms are in agreement with the advice to feed as soon as possible after parturition and to provide at least 6 liters within 24 h of age. The current study points at time of calving as a potential risk factor for sub-optimal colostrum feeding. Further research is necessary to determine the consequences of this observation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Robbers, Bijkerk, Koets, Benedictus, Nielen and Jorritsma.)

Random effects regression models are routinely used for clustered data in etiological and intervention research. However, in prediction models, the random effects are either neglected or conventionally substituted with zero for new clusters after model development. In this study, we applied a Bayesian prediction modelling method to the subclinical ketosis data previously collected by Van der Drift et al. (2012). Using a dataset of 118 randomly selected Dutch dairy farms participating in a regular milk recording system, the authors proposed a prediction model with milk measures as well as available test-day information as predictors for the diagnosis of subclinical ketosis in dairy cows. While their original model included random effects to correct for the clustering, the random effect term was removed for their final prediction model. With the Bayesian prediction modelling approach, we first used non-informative priors for the random effects for model development as well as for prediction. This approach was evaluated by comparing it to the original frequentist model. In addition, herd level expert opinion was elicited from a bovine health specialist using three different scales of precision and incorporated in the prediction as informative priors for the random effects, resulting in three more Bayesian prediction models. Results showed that the Bayesian approach could naturally take the clustering structure of clusters into account by keeping the random effects in the prediction model. Expert opinion could be explicitly combined with individual level data for prediction. However in this dataset, when elicited expert opinion was incorporated, little improvement was seen at the individual level as well as at the herd level. When the prediction models were applied to the 118 herds, at the individual cow level, with the original frequentist approach we obtained a sensitivity of 82.4% and a specificity of 83.8% at the optimal cutoff, while with the three Bayesian models with elicited expert opinion, we obtained sensitivities ranged from 78.7% to 84.6% and specificities ranged from 75.0% to 83.6%. At the herd level, 30 out of 118 within herd prevalences were correctly predicted by the original frequentist approach, and 31 to 44 herds were correctly predicted by the three Bayesian models with elicited expert opinion. Further investigation in expert opinion and distributional assumption for the random effects was carried out and discussed., Competing Interests: No authors have competing interests.

Clinical ketosis (CK) and subclinical ketosis (SCK) are associated with lower milk production, lower reproductive performance, an increased culling of cows and an increased probability of other disorders. Quantifying the costs related to ketosis will enable veterinarians and farmers to make more informed decisions regarding the prevention and treatment of the disease. The overall aim of this study was to estimate the combined costs of CK and SCK using assumptions and input variables from a typical Dutch context. A herd level dynamic stochastic simulation model was developed, simulating 385 herds with 130 cows each. In the default scenario there was a CK probability of almost 1% and a SCK probability of 11%. The herds under the no risk scenario had no CK and SCK, while the herds under the high-risk scenario had a doubled probability of CK and SCK compared to the default scenario. The results from the simulation model were used to estimate the annual cash flows of the herds, including the costs related to milk production losses, treatment, displaced abomasum, mastitis, calf management, culling and feed, as well as the returns from sales of milk and calves. The difference between the annual net cash flows of farms in the no risk scenario and the default scenario provides the estimate of the herd level costs of ketosis. Average herd level costs of ketosis (CK and SCK combined) were €3,613 per year for a default farm and €7,371 per year for a high-risk farm. The costs for a single CK case were on average €709 (with 5 and 95 percentiles of €64 and €1,196, respectively), while the costs for a single SCK case were on average €150 (with 5 and 95 percentiles of €18 and €422, respectively) for the default farms. The differences in costs between cases occurred due to differences between cases (e.g., cow culled vs cow not culled, getting another disease vs not getting another disease)., Competing Interests: The authors have declared that no competing interests exist.

Mammalian milk is not only a source of nutrition for the newborn, but also contains various components that regulate further development. For instance, milk is an abundant source of microRNAs (miRNAs), which are evolutionary conserved small non-coding RNAs that are involved in post-transcriptional regulation of target mRNA. MiRNAs present in milk can occur in extracellular vesicles (EVs), which are nanosized membrane vesicles released by many cell types as a means of intercellular communication. The membrane of EVs protects enclosed miRNAs from degradation and harbors molecules that allow specific targeting to recipient cells. Although several studies have investigated the miRNA content in milk EVs from individual species, little is known about the evolutionary conserved nature of EV-associated miRNAs among different species. In this study, we profiled the miRNA content of purified EVs from human and porcine milk. These data were compared to published studies on EVs from human, cow, porcine, and panda milk to assess the overlap in the top 20 most abundant miRNAs. Interestingly, several abundant miRNAs were shared between species (e.g., let-7 family members let-7a, let-7b, let-7f, and miR-148a). Moreover, these miRNAs have been implicated in immune-related functions and regulation of cell growth and signal transduction. The conservation of these miRNA among species, not only in their sequence homology, but also in their incorporation in milk EVs of several species, suggests that they are evolutionarily selected to regulate cell function in the newborn.

Societal concerns about antibiotic resistance prompted us to evaluate the use of prophylactic and postoperative antibiotic treatments following cesarean section (CS) and exploratory laparotomy (EL) in a surgery theater within the Farm Animal Health clinic. All procedures were performed by supervised veterinary students for training purposes. Cows undergoing CS (n = 100) received either a prophylaxis with a single 5-g injection of ampicillin-sodium i.v. only (CSL), or in combination with postoperative i.m. injections of ampicillin-trihydrate 20% for 3 d (10 mg/kg, twice a day; CSH). Cows undergoing EL (n = 110) received either no antibiotic prophylaxis (ELN) or were given a single 5-g injection of ampicillin-sodium i.v. (ELL). The primary outcome measure was healthy recovery after surgery, which we assessed according to the need to treat surgery-related complications within a 10-d follow-up period based on daily clinical observations. Cows in all groups had a normal temperature and feed intake, and a satisfying clinical appearance at the end of the follow-up period. The clinical need to treat cows with antibiotics to deal with postoperative complications was higher for the CS groups than the EL groups. Within both CS and EL groups, the number of complications for each protocol was the same. However, in terms of the secondary outcomes, we observed that CSL cows required, for example, more treatments for mastitis and other diseases unrelated to the surgery than CSH cows (odds ratio 2.8; confidence interval 1.2-7.2). The percentage of infected sutures was higher for ELN cows compared with ELL cows (odds ratio 2.6; confidence interval 1.5-4.9). We estimated that 29 CSH treatments were needed to prevent 1 CS cow with serious surgery-related complications in the CSL group. Likewise, 53 ELL treatments would prevent 1 EL cow with surgery-related complications in the ELN group. We therefore concluded that it is possible to reduce antibiotic prophylaxis in CS and EL cows. The low number cows of clinically detected complications were effectively treated with a postoperative antibiotic intervention at the moment of detection., (The Authors. Published by FASS Inc. and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).)

Previous reports have shown that reproductive tract score (RTS) can predict reproduction outcomes in seasonally bred beef heifers, although the accuracy can vary. Some ultrasonographic measures of the female reproductive tract and pelvis area have also been associated with reproductive outcome in young heifers. The objectives of this study were to determine which transrectal ultrasound or pelvis measures taken at a single examination are independent predictors of reproductive failure and whether the RTS system can be optimized with this information. In this observational study, year-old beef heifers (n = 488) in 2 birth cohorts were followed from just before the first breeding until confirmation of pregnancy. A single pre-breeding examination included body condition score, RTS, ultrasound measures of the reproductive tract (length and diameter of the left and right ovaries, presence and diameter of a CL, largest follicle diameter and left uterus horn diameter) and transverse and vertical diameters of the pelvis. Additional farm records including dam parity, sire, birth weight and birth date, weaning weight, weaning date, prebreeding body weight, AI dates, and semen used were available. Breeding consisted of 50 days of AI, followed 5 to 7 days later by a 42-day bull breeding period. Pregnancy failure was defined as the failure to become pregnant after the AI and bull breeding periods, while anestrus was defined as the failure to be detected in estrus during the 50-day AI period. From the prebreeding data and farm records, independent predictors of pregnancy failure and anestrus were identified using stepwise reduction in multiple logistic regression models. Age at the onset of breeding was the only consistent independent predictor of pregnancy failure and anestrus in both cohorts of this study (P < 0.05). Body condition score, uterus horn diameter, absence of a CL, largest follicle of less than 13 mm, and pelvis area (PA) were the prebreeding examination variables that remained in prognostic models (P < 0.1). Combining either the model based on the 3 remaining ultrasound measures or RTS with PA provided more accurate prognostic models for pregnancy failure and anestrus than using RTS alone (P < 0.05). It is concluded that ultrasound measures have prognostic value for pregnancy failure in restricted bred yearling heifers as a result of their association with anestrus, and that smaller PA has additional prognostic value for poorly performing heifers., (Copyright © 2016 Elsevier Inc. All rights reserved.)

This study investigated the metabolic effects of glucocorticoids when administered to propylene glycol-treated cows with clinical ketosis. Clinical ketosis was defined by depressed feed intake and milk production, and a maximal score for acetoacetate in urine. All cows received 250 mL oral propylene glycol twice daily for 3 days and were randomly assigned to a single intramuscular injection with sterile isotonic saline solution (n = 14) or dexamethasone-21-isonicotinate (n = 17). Metabolic blood variables were monitored for 6 days and adipose tissue variables for 3 days. β-Hydroxybutyrate (BHBA) concentrations in blood decreased in all cows during treatment, but were lower in glucocorticoid-treated cows. Cows treated with glucocorticoids had higher plasma glucose and insulin concentrations, whereas concentrations of non-esterified fatty acids, 3-methylhistidine and growth hormone were unaffected. mRNA expression of hormone-sensitive lipase, BHBA receptor and peroxisome proliferator-activated receptor type γ in adipose tissue was not affected. This shows that lipolytic effects do not appear to be important in ketotic cows when glucocorticoids are combined with PG. Plasma 3-methyl histidine concentrations were similar in both groups, suggesting that glucocorticoids did not increase muscle breakdown and that the greater rise in plasma glucose in glucocorticoid-treated cows may not be due to increased supply of glucogenic amino acids from muscle., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In a 7-year longitudinal study 292 Bovelder beef cows in a restricted breeding system in South Africa were observed from 1 to 2 days before their first breeding season, when reproductive tract scoring (RTS, scored from 1 to 5) was performed, until weaning their 5th calves. The objective was to determine whether pre-breeding RTS in heifers is a valid tool to predict long-term reproductive performance. Outcomes measured were failure to show oestrus during the first 24 days of the first 50-day AI season (24-day anoestrus), failure to become pregnant during each yearly artificial insemination (AI) season (reproductive failure), number of days from the start of each AI season to calving, and number of years to reproductive failure. The effect of RTS on each outcome was adjusted for year of birth, pre-breeding age, BW and body condition score (BCS), and for 24-day anoestrus, bull, gestation length, previous days to calving and previous cow efficiency index, the latter two in the case of the 2nd to the 5th calving season. During their first breeding season, heifers with RTS 1 and 2 combined were more likely to be in anoestrus for the first 24 days (OR=3.0, 95% CI 1.5, 6.4, P=0.003), and were also more likely to fail to become pregnant even after adjusting for 24-day anoestrus (OR=2.1, 95% CI 1.1, 3.9, P=0.025), compared to those with RTS 4 and 5 combined. Animals with RTS 1 and 2 combined were at increased risk of early reproductive failure compared to those with RTS 4 and 5 combined (HR=1.4, 95% CI 1.0, 1.9, P=0.045) although RTS was not associated with calving rate or days to calving after the second calving season. Low RTS at a threshold of 1 had consistent specificity of ≥94% for both 24-day anoestrus and pregnancy failure, however its predictive value was lower in the age cohort with a higher prevalence of anoestrus. We conclude that RTS is a valid management tool for culling decisions intended to improve long-term reproductive success in a seasonal breeding system, by excluding heifers that are likely to fail to become pregnant or likely to calve late during their first calving season., (Copyright © 2014 Elsevier B.V. All rights reserved.)

An in vitro model was used to investigate effects of β-hydroxybutyrate and isoproterenol (β-adrenergic receptor agonist) on lipolysis in isolated adipocytes from late pregnant and recently calved dairy cows (n=5) and cows with clinical ketosis (n=3). Incubation with 3.0 mmol/L β-hydroxybutyrate reduced lipolysis in isolated adipocytes. This inhibitory effect was lower in the first lactation week (47%±16%) compared with late pregnancy (71%±6.5%). Incubation with 0.3 μmol/L isoproterenol stimulated lipolysis in isolated adipocytes from periparturient dairy cows. Basal lipolysis resulted in non-esterified fatty acid to glycerol ratios in the incubation media of 2.0±0.23 in prepartum samples, 2.1±0.23 in the first lactation week and 2.2±0.09 in cows with clinical ketosis. β-Hydroxybutyrate reduced lipolysis by 45%±9.6% in isolated adipocytes from cows with clinical ketosis, indicating that impaired feedback of β-hydroxybutyrate may not play a role in the disease etiology., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

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., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The conception rate in dairy cows is dependent on a number of cow factors such as days in milk and insemination number. Unfortunately, some of these factors were not accounted for in optimal insemination and replacement decision models. By using wrong estimates of the conception rate, the calculated optimal insemination and replacement policy might differ from the real optimal insemination and replacement policy. The objective of this study was to evaluate different sets of conception rates with an increasing level of accuracy to determine the insemination policy. An existing dynamic program for optimal insemination and replacement was used to compare three different scenarios in the estimation of conception rates, based on the reproductive performance of Dutch dairy cattle: (i) constant conception rate throughout lactation, (ii) conception rate dependent on parity and months in milk, and (iii) conception rate dependent on parity, months in milk and insemination number. The time step of the model was 30.4 days (1 month). The discounted future cash flow of culling a cow at each time step (replace a heifer immediately) was compared with keeping that cow under optimal future decisions. The difference between immediate culling and optimal decisions is defined as the retention pay-off. The insemination value was calculated as the difference between the future cash flow between immediate insemination of a cow and waiting one time-step. The results show that the difference in the insemination values and the optimal time to stop insemination depend on parity, lactation stage and the relative milk yield. In older cows with equal milk yields and at the same months in milk, the insemination value was lower than in younger cows. Within a parity, the insemination value was higher for cows with a higher milk yield. On individual cow level, using more accurate conception rate as input in the optimal insemination and replacement model might reduce miscalculation of the economic consequences for at least of €20-€38 per cow per year. Basing insemination decisions on less accurate input of the probabilities of conception, however, did not have an economic consequence at the herd level. In conclusion, using the appropriate conception rate as input in the optimal insemination and replacement model would increase the precise decision for the optimal time to stop insemination and hence improve the reproductive management efficacy., (© 2012 Blackwell Verlag GmbH.)

The objective of this study was to obtain information on variation between dairy cows in muscle and fat tissue mobilization around parturition and to study the association between protein and fat mobilization and serum β-hydroxybutyrate (BHBA) concentrations (hyperketonemia) in this period. Thirty-four cows kept under similar conditions at a university dairy farm (no experimental treatments) were monitored from 4 wk before until 8 wk after calving. Mobilization of muscle protein was investigated by analysis of plasma 3-methylhistidine concentrations (3-MH, analyzed by a recently developed HPLC tandem mass spectrometry method) and ultrasound measurements of longissimus muscle thickness. Mobilization of fat tissue was monitored by serum nonesterified fatty acid (NEFA) concentrations and ultrasound measurements of backfat thickness. Large variation was observed between cows in onset and duration of periparturient protein and fat mobilization. Plasma 3-MH concentrations and muscle thickness profiles indicated that protein mobilization started, on average, before parturition and continued until approximately wk 4 of lactation. Serum NEFA concentrations and backfat thickness profiles showed that fat mobilization occurred from parturition until the end of the study. Thus, muscle protein mobilization occurred in advance of fat mobilization in most cows from this study. We hypothesized that this might be due to a prepartum amino acid deficiency in the absence of negative energy balance. The incidence of hyperketonemia in this study was 16/34 = 47%. With the exception of 3 cows defined as having severe hyperketonemia, cows with lower 3-MH concentrations had higher serum BHBA concentrations. A possible explanation for this observation might be that higher mobilization of protein around calving might restrict ketone body production due to the higher availability of glucogenic precursors in the period of most severe negative energy balance and highest fat mobilization. The validity of this hypothesis needs to be confirmed, but data from this study indicate that further research on the role of protein mobilization in the etiology of hyperketonemia in dairy cows is needed., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

To improve monitoring of protein mobilization in dairy cows, we developed and evaluated a method to quantify 1-methylhistidine and 3-methylhistidine in plasma by HPLC-tandem mass spectrometry. The analytical method described is (1) sensitive: both histidine derivates can be detected in the picomole range; (2) accurate: intra- and interassay coefficients of variation were < 5% for all standard solutions of 1-methylhistidine and 3-methylhistidine measured (31 to 500 pmol); (3) specific: 1-methylhistidine is clearly separated from 3-methyl-histidine in plasma samples from dairy cows; and (4) flexible: can be easily adapted to measure other amino acids or compounds containing a primary amine. 1-Methylhistidine is present in plasma of dairy cows at concentrations of 5.0 ± 1.7 μM, similar to concentrations of 3-methylhistidine (4.4 ± 2.4 μM). Analytical separation of both histidine metabolites is essential when plasma 3-methylhistidine is used as indicator for muscle breakdown in dairy cows. Specific quantification of the concentration of 3-methylhistidine in bovine plasma samples by HPLC tandem mass spectrometry can improve monitoring of protein mobilization in dairy cows., (Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

Problem: In cattle, retained placenta (RP) is suggested to arise from failure of immune-mediated rejection of the fetal membranes by the maternal immune system and is associated with major histocompatibility (MHC) class I compatibility between calf and dam., Method of Study: To study the association between RP and different MHC class I compatibilities between calf-dam-granddam combinations, massively parallel pyrosequencing was used to determine the MHC class I haplotypes of cows with and without RP., Results: Two-way calf to dam MHC class I compatibility gave a high risk for RP. There was a tendency for a higher risk for RP with calf to dam MHC class I compatibility., Conclusions: We concluded that in two-way compatible pregnancies, the maternal immune system fails to reject the fetal membranes, and the fetal immune system does not mount an immune response against maternal MHC class I antigens that could influence the immune-mediated rejection of the fetal membranes by the maternal immune system. The lack of immune-mediated rejection of the fetal membranes by the maternal immune system increases the risk of occurrence of RP., (© 2011 John Wiley & Sons A/S.)