Adult Dairy Cattle Research Articles

Enhancing dairy cow population through decreasing calf mortality and increasing calving rate in Indonesia: a system dynamics approach

Abstract The dairy industry in Indonesia is facing significant challenges due to the rates of calf mortality and low calving rates. Calf mortality and calving rate became critical factors that influence the sustainability of the dairy cow population. This study aimed to assess reproduction management by decreasing calf mortality and increasing calving rates to enhance the dairy cow population in Indonesia. This study employed secondary data in 2022 using technical parameter coefficients from previous research. A causal loop diagram and a simulation model using a system dynamics approach were developed to investigate the dynamics of the dairy cow population under different scenarios of calf mortality and calving rates from 2023 to 2030. The results showed that decreasing calf mortality by 2% may increase dairy cow population by 1,991 head/year (0.31%), heifer population by 417 head/year (0.63%), and adult female dairy cattle population by 389 head/year (0.09%). The finding of this study also showed that increasing calving rate by 5% may increase dairy cow population by 17,854 head/year (2.75%), heifer population by 3,698 head/year (5.56%), and adult female dairy cattle population by 2,137 head/year (0.51%). It is concluded that reducing calf mortality rates and improving calving rates should be prioritized to enhance the dairy cow population in Indonesia.

Clostridioides difficile in calves, cattle and humans from Dutch dairy farms: Predominance of PCR ribotype 695 (clade 5, sequence type 11) in cattle

BackgroundClostridioides difficile is a leading cause of infectious diarrhea in both humans and livestock. In particular, C. difficile strains belonging to sequence type (ST) 11 are common enteropathogens. The aim of this study was to determine the presence and genetic relatedness of C. difficile types in dairy cattle and calves. MethodDutch dairy farms were visited between February and December 2021. Feces was collected from adult dairy cattle and calves of two age categories (<4 weeks and 4 weeks-4 months). Fecal samples were also requested from dairy farmers, family members and employees. Fecal samples were cultured in an enrichment medium for 10–15 days and subcultured on solid media for capillary PCR ribotyping and whole genome sequencing. ResultsC. difficile was detected on 31 out of 157 (19.8%) dairy farms. The highest prevalence was found in calves <4 weeks (17.5%). None of the 99 human samples collected were positive. Thirty-seven cultured isolates belonged to 11 different PCR ribotypes (RT) of which RT695 (56.8%) and RT078/126 (16.2%) were most abundant. In the database of the Netherlands National Expertise Centre for C. difficile infections (CDI, >10.000 patient isolates), RT695 was found in only two patients with hospital-onset CDI, diagnosed in 2020 and 2021. Sequence analysis of 21C. difficile RT695 from cattle revealed that all isolates belonged to clade 5, ST11 and contained genes encoding toxin A, toxin B and binary toxin. RT695 strains carried antimicrobial resistance genes typically found in clade 5C. difficile. Groups of genetically related RT695 isolates were found between dairy farms, whereas identical strains were only present in individual farms. ConclusionsC. difficile was found in ∼20% of dairy farms with a predominance of the relatively unknown RT695. Isolates of RT695 belonged to the same clade and sequence type as RT078/126, which is recognized as an important zoonotic type.

Actively ventilating calf hutches using solar-powered fans: Effects on hutch microclimate and calf thermoregulation

Although active ventilation via fans is an effective and widely adopted heat abatement method for use with adult dairy cattle, it has yet to be investigated in outdoor hutch-housed dairy calves despite most US calves being raised in such systems. We investigated a solar-powered fan system for outdoor calf hutches and its effect on hutch microclimate and calf thermoregulation. During summer, a 3 × 3 Latin square was replicated 4 times (n = 12 preweaning heifers) with 4-d exposure periods to minimally (CON; rear windows closed), passively (PASS; rear windows opened), or actively (ACT; solar-powered fan, activated at dry bulb temperature [Tdb] > 21°C) ventilated hutch systems. Hutch microclimate and calf thermoregulation were evaluated either continuously (Tdb, humidity, rectum surface temperature, and behavior) or after a daily 30-min inside restriction (air speed, air particle number, noise level, respiration, and sweating rate, and skin and rectal temperature). Active ventilation substantially increased hutch air speed relative to PASS and CON (1.76 vs. 0.19 vs. 0.05 m/s). However, PASS hutches had the lowest INT Tdb (27.2 vs. 26.4 vs. 27.8°C), whereas ACT INT Tdb was reduced at 0900 and 1000 h relative to CON but not PASS. Similarly, ACT reduced calf respiration rates and lowered rectum surface temperature at 0800 and 0900 h when compared with CON but not PASS. The lack of strong ACT influence on calf outcomes over PASS could partially be explained by the decreased proportion of time ACT calves spent inside their hutch (48.7 vs. 67.3 vs. 64.1% of each hour). Overall, ACT improved hutch microclimate and calf responses relative to CON but not PASS. Either ACT or PASS ventilation may be sufficient to provide heat abatement to continental hutch-housed calves.

Development and implementation of an onfarm animal health and diagnostic educational training program for farm workers in antimicrobial stewardship in adult dairy cattle

Antimicrobial stewardship (AMS) is a holistic approach, which promotes judicious use of antimicrobial drugs to preserve their effectiveness and availability. Dairy farm workers are com­monly responsible for disease diagnosis and routine treatment decisions. This highlights the importance of farm workers’ knowledge and skills to successfully implement judicious use of antimicrobials in livestock production systems. Knowledge gaps in animal health and behavior have been recognized be­fore as an important reason to provide training to farm work­ers. A better understanding of cattle sickness behavior will improve early identification of clinical signs of disease and im­prove treatment success. The main objective of this project was to develop and implement an on-farm educational training pro­gram for farm workers in antimicrobial stewardship in adult dairy cattle.

Antimicrobial stewardship on the dairy: Evaluating an on-farm framework for training farmworkers

Dairy farmworkers are commonly responsible for disease diagnosis and routine treatment decisions for cattle. This highlights the importance of farmworkers' knowledge and skills to successfully implement judicious use of antimicrobials in livestock production systems. The main objectives of this project were to develop and evaluate an on-farm educational program for farmworkers in antimicrobial stewardship in adult dairy cattle. A longitudinal quasi-experimental study design was used, by enrolling 12 conventional dairy farms in the United States (6 in California and 6 in Ohio). Farmworkers responsible for treatment decisions on the farm (n = 25) participated in a didactic and hands-on 12-wk antimicrobial stewardship training program led by the investigators. All antimicrobial stewardship training materials were available in Spanish and English. Interactive short videos with audio were developed to cover the learning objectives for each of the 6 teaching modules: antimicrobial resistance, treatment protocols, visual identification of sick animals, clinical mastitis, puerperal metritis, and lameness. Pre- and post-training assessments were administered using an online training assessment tool to evaluate changes in knowledge and attitudes about antimicrobial stewardship practices. Cluster analysis and multiple correspondence analyses were conducted to evaluate the association among categorical variables for participants' level of change in knowledge and its association with language, farm size, and state. A 32% average increase in knowledge was observed through an assessment conducted after completing the antimicrobial stewardship training, compared with the pre-training assessment. A significant improvement in 7 of 13 attitude questions related to antimicrobial stewardship practices on the farm was observed. Knowledge and attitude scores of participants on antimicrobial stewardship and identification of sick animals significantly improved after completing the antimicrobial stewardship training. The results observed in this study support the relevance of antimicrobial stewardship training programs targeting farmworkers to improve antimicrobial drug use knowledge and skills.

Fecal cortisol metabolites reflect transport stress in 3-month-old dairy calves pre- and postweaning: A pilot study

Measurement of fecal cortisol metabolites (FGCM) is a well-established, noninvasive method to assess stress in adult dairy cattle. However, this procedure has not yet been validated for unweaned dairy calves, and it can be expected that the milk proportion of the diet may influence the resulting FGCM concentrations. The aim of this study was therefore to assess whether a peak in FGCM concentrations in response to a stressor can be measured in unweaned dairy calves on a largely milk-based diet. If so, further objectives were to examine whether maximum FGCM concentrations, as well as the time lag until they are reached, are comparable to the values in the same calves on a solid-based diet after weaning. For this study, 5 German Holstein calves of about 3 mo of age (93 to 102 d preweaning) were exposed to a 45 min transport stressor once before and once after weaning, which was 3 wk apart. All voided fecal samples were collected for 24 h after termination of the transport. Fecal cortisol metabolites were analyzed with an 11-oxoetiocholanolone enzyme immunoassay and changes in FGCM concentrations relative to the individual baseline (FGCMrel) were calculated. Results showed a clear peak in FGCM concentrations on both diet types. The peak FGCMrel concentrations tended to be higher when the calves were on the preweaning diet (at peak: +233 ± 25% increase relative to baseline) in comparison to the postweaning diet (+124 ± 23%). Considering the whole 24 h sampling period, the FGCMrel concentrations for all calves were significantly higher on the preweaning diet than on the postweaning diet. There was also a numerical difference in the delay between occurrence of the stressor and appearance of the peak FGCMrel concentrations in feces, as the time lag was 1.5 ± 1.2 h longer when the calves were on the preweaning diet compared with the postweaning diet. In conclusion, our results suggest that FGCM concentrations are a useful stress marker for unweaned dairy calves in the same way they are for older cattle, but that FGCMrel concentrations tend to be higher in unweaned than in weaned calves and are thus not directly comparable.

Distinct Mannheimia haemolytica serotypes isolated from fatal infections in veal calves and dairy cows

Fatal Mannheimia haemolytica (M. haemolytica) infections in cattle, which emerged in the Netherlands between 2004 and 2018, showed two distinct disease presentations: acute fibrinous polyserositis (FPS) in veal calves, and acute fibrinous pleuro-pneumonia (FPP) in adult dairy cattle. To determine whether these presentations were caused by different M. haemolytica genotypes, whole genome sequencing was performed on 96 isolates cultured after necropsy from inflamed sites of veal calves that died of M. haemolytica-associated FPS (n = 49) or with FPP lesions (n = 2), and from dairy cows that died of M. haemolytica-associated FPP (n = 45). Among the 96 M. haemolytica isolates, 93 were shown to belong to either of two large clusters, with 48/51 calf isolates belonging to one, and 43/45 cow isolates and two calf isolates from cases of FPP to the other. All M. haemolytica isolates from veal calves with FPS were of serotype A2, whereas the isolates from dairy cows and two calves with FPP were predominantly of serotypes A1 and A6. Most serotype A2 isolates from veal calves with FPS (95.6 %) contained multiple antibiotic resistance genes (ARGs) against three to five antimicrobial classes (phenicols, sulphonamides, tetracyclines, aminoglycosides or beta-lactams). In contrast, these ARGs were only present in 10.8 % of M. haemolytica A1 and A6 isolates from pneumonic adult cattle and absent in isolates from the two calves with FPP. These two disease presentations appear to be caused by genetically distinct strains with different antimicrobial resistance gene patterns. While M. haemolytica serotype A2 is generally considered to be a commensal microorganism of cattle, it was clearly associated with fatal FPS in veal calves in the Netherlands.

Molecular identification of Cryptosporidium species in Canadian post-weaned calves and adult dairy cattle.

Cryptosporidium is a zoonotic protozoan parasite that is distributed globally and impacts both human and animal health. There are over 40 species of Cryptosporidium described to date, of which four (C. parvum, C. bovis, C. ryanae and C. andersoni) are routinely reported in cattle. The goal of this study was to identify the Cryptosporidium species infecting dairy cattle from across Canada using cow fecal samples and post-weaned calf rectal swabs obtained through the Canadian National Dairy Study. A total of 353 cattle samples (117 pooled rectal fecal swabs from post-weaned calves and 236 cow fecal samples) from 175 herds across the 10 Canadian provinces were analysed by targeting Cryptosporidium's small subunit ribosomal RNA (SSU rRNA or 18S) gene. Herd prevalence of Cryptosporidium was 27.4% nationally, ranging from 0% in Saskatchewan (SK) to 62% in Prince Edward Island (PE). The national prevalence of Cryptosporidium cattle infections was 15.4% in pooled rectal fecal swab samples from post-weaned calves and 16.1% in adult cows. Sanger sequence analysis of the SSU rRNA gene target revealed that C. bovis, C. andersoni and C. ryanae occurred in both adults and post-weaned calves, with C. bovis as the predominant species detected in pooled fecal swab samples of post-weaned calves (9/18, 50%) and C. andersoni as the predominant species in cows (25/38, 66%). Cryptosporidium parvum was not observed in any of the pooled rectal swab samples from post-weaned calves but was observed in one mixed infection of C. bovis/C. parvum in an adult cow. The fifth species identified in this study was C. muris and was present in two adult cows. Low concentrations of oocyst equivalents, as measured by quantitative real-time PCR (qPCR) of the SSU rRNA gene copy number, were observed in a subset of cattle samples. Cryptosporidium andersoni concentrations varied from province to province, with the widest range and highest counts in cows from PE. In conclusion, oocysts from Cryptosporidium species observed in this study are shed into the environment, contributing to the environmental load. However, the Cryptosporidium species in the post-weaned calves and cows found in this study pose a lower risk to the dairy cattle than if they were infected with C. parvum. Similarly, these Cryptosporidium species pose only a small risk to public health as the three species are infrequently reported in humans globally and have not been reported in Canadians to date.

Clinical examination of cattle. Part 2: calves, technology and ancillary testing

Background: A systematic clinical examination of calves is key for reaching a diagnosis and creating a management plan for sick animals, with the principles of an individual animal clinical examination giving an indication of group-level issues within the herd. The use of ancillary testing can aid in diagnosing different conditions in both youngstock and adult cattle, as well as determining the prognosis for the animal. This has become more accessible recently due to the developments in sensors and the application of diagnostic imaging, as well as calf-side tests. Aim of the article: This article is the second in a two-part series on undertaking a clinical examination of cattle. This article focuses on how to perform a clinical examination of a calf, at both an individual and a population level, and also discusses the role of technology and ancillary testing in aiding the clinical examination of both adult cattle and calves. Part 1 focused on examining adult dairy and beef cattle (Nelson and others 2022).

Investigation of Weather Triggers Preceding Outbreaks of Acute Bovine Liver Disease in Australia.

Acute bovine liver disease (ABLD) is a hepatic disease affecting cattle sporadically in southern Australia, characterised histologically by striking periportal hepatocellular necrosis. The cause of ABLD is unknown; however, the seasonality and acute presentation of outbreaks suggest mycotoxin involvement. We described the geographical and seasonal occurrence of ABLD reports from 2010 to 2020 in Victoria, Australia, and explored potential weather triggers preceding 26 outbreaks occurring across 23 properties using a case-crossover design. Outbreaks occurred most frequently in autumn/early winter and in herds located along the southern coastal plain of Victoria, and occasionally within the low-lying regions of the Great Dividing Range. Lactating adult dairy cattle represented the most reported cases. We observed a significant association between an increase in average daily dewpoint in the 15 days preceding an ABLD outbreak, suggesting that dew formation may be a key determinant for this disease. Our findings support the etiology of a potent hepatotoxic agent that requires moisture for proliferation and/or toxin production.

Clinical examination of cattle. Part 1: Adult dairy and beef cattle

Background: Carrying out a thorough and systematic clinical examination is a key veterinary skill. The high value of individual animals, and the sometimes limited access to further diagnostics in the field make this particularly important when working with cattle. Cattle may require a clinical examination after presenting as unwell to reach a diagnosis or to aid development of a management plan. The principles of an individual clinical examination can also be applied to population medicine. Individual animal findings may highlight herd‐level risk factors and warrant further investigation. Additionally, a population subset of animals can undergo a targeted examination to determine prevalence of disease in a herd. Finally, a group of animals can be examined as a whole to identify behavioural signs indicative of a problem. At a population level it is important to remember to also thoroughly look at the environment and to examine both farm records and herd data.Aim of the article: In this first article, we will outline an approach to the examination of adult cattle at both an individual and population level. A follow‐up article, to be published in a subsequent issue, will cover examination of youngstock and the use of technology and ancillary testing.

Preweaning dairy calves' preferences for outdoor access

Adult dairy cattle show a preference for outdoor spaces during summer nights, but little is known about such preferences for dairy calves. Our aim was to determine the preferences of dairy calves for outdoor access during their first 11 wk of age in summer conditions. Calves were paired (n = 10) at 7 d of age and placed in one of 10 pens (7.32 × 2.44 m, deep bedded with bark mulch), each with equally sized outdoor and indoor areas. All feed was provided indoors; calves had ad libitum access to water, hay and concentrate. During the preweaning period, from 5 to 29 d of age, calves were offered 12 L/d of milk divided into 2 feedings. Calves were weaned stepwise, with milk gradually reduced by 1 L/d from d 30 to 35 of age. Calves were fed 6 L/d from 35 to 51 d of age, then milk was again reduced by 1 L/d so that calves were completely weaned by 56 d of age. Calves remained in the trial until, on average (± SD), 72 ± 2 d of age. Pens were continuously video recorded, and behaviors were scored 3 d/wk using 5-min scan samples. Four observers scored whether calves were inside, outside, lying, standing, in the sun or shade (interobserver reliability = 0.97). Sun and shade could not always be detected, so a proportion was calculated as the amount of time in the sun or shade divided by the total time sun or shade was visible. Calves spent approximately one-third of their time outside throughout the experiment, and time spent outside was affected by several factors. Pairs of calves spent less time outside as the amount of rain increased. During the first 6 wk of age, time spent outside increased, but time spent outside decreased between wk 6 and 7, and remained around 200 min/d from wk 7 to 11. Calves with the highest average daily gain and calves that were approximately 50% white appeared to spend more time outdoors. When outside, calves spent similar amounts of time in the sun and shade during the first 8 wk, but after weaning they appeared to spend more time in the shade. When calves were outside they appeared to spend a greater proportion of their time standing than when they were indoors during the first 4 wk. These results indicate that, when given the option in the summer, calves make use of an outdoor space, but this appears to vary with weather, calf age, average daily gain, and coat color.

Detection of Antibodies Brucella abortus in dairy cattle in Puspo District, Pasuruan using Rose Bengal Test and Complement Fixation Test

Brucellosis was an infectious disease caused by the genus brucella. Brucellosis in dairy cattle was caused by Brucella abortus that impaction an abortion. The aim of this study was to determine the presence of antibodies of the Brucella abortus in adult female dairy cattle in the Puspo district Pasuruan using Rose Bengal Test (RBT) and Complement Fixation Test (CFT). A descriptive analysis method with quantitative approach was used in this study. Meanwhile, interviews were conducted to get supporting information from the farmer. Used 100 samples. The results of the study showed 2% samples considered positive brucellosis tested by RBT. To confidence false-positive result, RBT test was followed by CFT. After tested by CFT, in this study there were no brucella antibodies in adult female dairy cattle in Puspo district, Pasuruan.

Prevalence and Risk Factors of Bovine Mastitis in Southern Zone of Tigray, Northern Ethiopia.

Mastitis is the most common disease of adult dairy cattle. The disease continues to be one of the most perplexing and costly diseases of dairy cattle. The present study was conducted to detect bovine mastitis cows and identify potential risk factors for the distribution of bovine mastitis of smallholder dairy farmers using the California mastitis test. A cross-sectional study was conducted from September 2013 to May 2015 in the southern zone of Tigray, Northern Ethiopia, on 146 head of randomly selected cattle, of which 99 (67.8%) were crossed zebu and 47 (32.2%) pure local zebu using the California mastitis test and questionnaire. The overall prevalence was 65 (45.5%), of which 72.3% of crossed local and 27.69% of local zebu were found positive for the CMT test. The investigated risk factors were the season of lactation (OR = 0.510, CI = 0.208–1.247, P=0.140), washing hands between consecutive milking events (OR = 0.354, CI = 0.146–0.856, P=0.021), and history of previous mastitis (OR = 0.441, CI = 0.226–0.858, P=0.016), which had significant (P < 0.05) effect on the present prevalence of bovine mastitis in the study area. A high prevalence of bovine mastitis was observed in the southern zone of Tigray. The summer season of lactation and not washing hands between consecutive milking of cows were significant mastitis causation risk factors in addition to the history of previous mastitis disease history.

Characterization of Winter Dysentery Bovine Coronavirus Isolated from Cattle in Israel.

Bovine coronavirus (BCoV) is the causative agent of winter dysentery (WD). In adult dairy cattle, WD is characterized by hemorrhagic diarrhea and a reduction in milk production. Therefore, WD leads to significant economic losses in dairy farms. In this study, we aimed to isolate and characterize local BCoV strains. BCoV positive samples, collected during 2017–2021, were used to amplify and sequence the S1 domain of S glycoprotein and the full hemagglutinin esterase gene. Based on our molecular analysis, local strains belong to different genetic variants circulating in dairy farms in Israel. Phylogenetic analysis revealed that all local strains clustered together and in proximity to other BCoV circulating in the area. Additionally, we found that local strains are genetically distant from the reference enteric strain Mebus. To our knowledge, this is the first report providing molecular data on BCoV circulating in Israel.

124 Quantifying temperament using behavioural testing in adult dairy cattle

124 Quantifying temperament using behavioural testing in adult dairy cattle

A history of lameness and low body condition score is associated with reduced digital cushion volume, measured by magnetic resonance imaging, in dairy cattle

Claw horn lesions (CHL) are the result of a failing of the functional anatomy of the hoof in dairy cows. The digital cushion is understood to be a vital structure in the prevention of CHL. Claw horn lesions have previously been shown to lead to pathological change to the pedal bone; however, their effects on the digital cushion are unknown. The primary aim of this study was to examine associations between the history of CHL through an animal's life and the structure of the digital cushion at slaughter using magnetic resonance imaging. The retrospective cohort study resulted in the scanning of 102 pairs of hindfeet, collected from adult Holstein dairy cows culled from a research herd, using a 3-Tesla research-grade magnetic resonance imaging scanner. Volume and fat measurements were calculated for each digital cushion within each claw from a modified Dixon Quant sequence. Animal-level variables were constructed around the animals' lactating lifetime, with lameness scores and body condition score collected at least every 2 wk. The combined volume of digital cushion in the lateral claws was used as the outcome variable in multivariable linear models. The volume of the digital cushion was negatively associated with the number of lameness events or CHL recorded. Furthermore, animals with body condition score >3, culled later in lactation, or of a greater body weight were more likely to have a higher volume of digital cushion in the lateral claws. We propose that the observations made in the current study are the effects of a range of factors broadly associated with genetic, developmental, and disease-related inputs. Our understanding of how we can select for genetically more robust animals and how we can precondition the hoof before first calving needs to be improved to reduce the risk of future CHL in adult dairy cattle. Furthermore, understanding optimal treatment regimens and their effect on hoof anatomy may reduce the recurrence of CHL in the current lactation and future lactations.

English

The current study was planned to evaluate the transplacental transmission potentials of Anaplasma marginale among naturally infected adult dairy cattle in district Jhang, Punjab, Pakistan. A dairy farm was selected after a district level survey having highest number of blood smear, cELISA and PCR positives of A. marginale infected pregnant crossbred cows (Holstein Friesian × Cholistani). Blood samples were collected from dams (within 4 months of their pregnancies and at the time of parturitions) as well as from the new-born calves (before colostrum feeding within 3 h of the birth). Eighty pregnant cows were screened through the cELISA and PCR. Out of these, 54 were found positive for anaplasmosis. Further follow up revealed that 38 cows remained positive at the time of parturition. Finally, based on the criteria, 32 calves without colostrum feeding qualified for the vertical transmission trial. The results of present study revealed that 28 (9/32) and 13% (4/32) of the neonatal calves acquired intrauterine infection confirmed by cELISA and PCR, respectively. Overall occurrence of the transplacental transmission was 31%. Nevertheless, we concluded that transplacental transmission occurs and this route of transmission, can lead to a significant number of neonatal deaths. Hence, the transplacental route of disease transmission should also be considered for devising the prevention and control strategies regarding anaplasmosis in the dairy cattle. © 2021 Friends Science Publishers

Melatonin enhances in vitro developmental competence of cumulus-oocyte complexes collected by ovum pick-up in prepubertal and adult dairy cattle

Bovine oocytes from prepubertal donors have been used for in vitro embryo production to decrease the generation interval. However, reduced cumulus-oocyte competence, mainly attributed to increased apoptosis, has been observed in oocytes/embryos collected from prepubertal donors. Here, we investigated the effects of the potent antioxidative molecule melatonin on cumulus-oocyte competence and embryo development in prepubertal and adult dairy cattle in vitro. A total of fifteen Holstein Friesian calves, six to ten months old (7.6 ± 1.34 months of age). And fifteen adult cows with one to four calvings (2.3 ± 0.96 calvings) were enrolled as ovum pick up (OPU) donors in this study. Cumulus-oocyte complexes (COCs) were cultured either in the presence or absence of melatonin (0.01 nM). The proportion of cleavage stages, blastocysts, and advanced blastocysts was determined. Embryo quality was assessed via differential staining to determine the total embryonic cells and allocation to the inner cell mass (ICM) and trophectoderm (TE) cells. Melatonin treatment yielded a greater percentage of blastocysts compared to the control group, i.e. oocytes from both adult cows (P = 0.0485; 24.8 ± 3.5% vs. 16.0 ± 3.4%, respectively), and from prepubertal donors (P = 0.0007; Melatonin 23.1 ± 5.1% vs. Control: 11.1 ± 3.5%). Adult cows had significantly (P = 0.0370) greater advanced blastocyst rates than those found in the prepubertal group (13.9%± vs. 7.0±%, respectively). Additionally, the number of ICM, total cells, and the ratios ICM: Total, ICM: TE, respectively, were greater (P < 0.05) after melatonin treatment compared with the control group (39.1 ± 2.8, 98.6 ± 5.7, 0.4 ± 0.01, and 0.7 ± 0.04 vs. 27.3 ± 2.9, 81.2 ± 5.8, 0.34 ± 0.01, and 0.52 ± 0.04, respectively). Blastocysts derived from adult cows had a greater number of TE (P = 0.01) and total embryonic cells (P = 0.0095) compared to the prepubertal donor group (63.5 ± 3.2 and 101.05 ± 4.8 vs. 48.9 ± 4.3 and 78.8 ± 6.5, respectively). Nevertheless, embryonic cell counting in embryos derived from prepubertal COCs equated to that observed from adult donors after melatonin exposure. In conclusion, these results indicate that the presence of melatonin during in vitro maturation improves cumulus-oocyte competence, embryo development, and quality by increasing the allocation of embryonic cells to the ICM compartment and the total number of embryonic cells in both adult and prepubertal bovine donors.

Estimation of the sensitivity and specificity of four serum ELISA and one fecal PCR for diagnosis of paratuberculosis in adult dairy cattle in New Zealand using Bayesian latent class analysis

In New Zealand, a new diagnostic approach for the control of paratuberculosis in mixed aged milking cows has been developed using a combination of ELISA and quantitative fecal PCR (f-qPCR). Our analysis was designed to evaluate performance of these individual tests in infected or infectious mixed aged cows across the prevalence of infection typically encountered on NZ dairy farms and calculate test accuracy when used as a screening test of serological ELISAs for four separate antigens read in parallel followed by a confirmatory quantitative f-qPCR test.Data from a cross-sectional study of 20 moderate prevalence herds was combined with existing data from 2 low and 20 high prevalence herds forming a dataset of 3845 paired serum and fecal samples. Incidence of clinical Johne’s disease (JD) was used to classify herds into three prevalence categories. High (≥ 3% annual clinical JD for the last three years), moderate (<3 – 1%) and low (<1% incidence for at least the last five years). Positive tests were declared if> 50 ELISA units and f-qPCR at two cut-points (≥1 × 104 genomes/mL or >1 × 103 genomes/mL).Fixed Bayesian latent class models at both f-qPCR cut-points, accounted for conditional independence and paired conditional dependence. Mixed models at both f-qPCR cut-points, using a different mechanism to account for conditional dependencies between tests were also implemented. Models (24 in number) were constructed using OpenBUGS. The aim was to identify Mycobacterium avium subsp. paratuberculosis (MAP) infected cows that met at least one of two criteria: shedding sufficient MAP in feces to be detected by f-qPCR or mounting a detectable MAP antibody response.The best fit to the data was obtained by modelling pairwise dependencies between tests in a fixed model or by accounting for dependencies in a mixed model at a fecal cut-off of ≥1 × 104 genomes/mL. Test performance differed with prevalence, but models were robust to prior assumptions. For the fixed model, at a prevalence of 0.29 (95 % probability interval (PI) = 0.25−0.33), as a screening plus confirmatory f-qPCR, post-test probability for disease in a positive animal was 0.84 (95 %PI = 0.80−0.88) and 0.16 (95 %PI = 0.15−0.18) for disease in a test negative animal. In low prevalence herds (0.01(95 %PI = 0.00−0.04)) the equivalent figures were 0.84 (95 %PI = 0.08−0.92) and 0.00 (95 %PI = 0.00−0.02).These results suggest this is a useful tool to control JD on dairy farms, particularly in herds with higher levels of infection, where the sampling and testing cost per animal is defrayed across more detected animals.