Evaluation of selected risk factors for different stages of digital dermatitis in Dutch dairy cows.

Digital dermatitis (DD) is a painful infectious disease in dairy cattle that causes ulcerative lesions of the skin just above the coronary band, mainly of the hind legs. Estimates for DD prevalence at cow level in the Netherlands range from 20% to 25%. In this study, risk factors for the various stages of DD were identified and quantified. The hind legs of 6766 cows on 88 farms were scored by trained interns, using the M-scoring system (M0-M4.1). Farms in this study were a convenience sample, based on the prevalence of DD as recorded at the latest herd trim, geographical location and willingness of the farmers to participate. A survey with questions about cow environment and herd management was conducted by the intern at the day of scoring. The data were collected between August 2017 and January 2018. DD was found on 38.6% of the scored legs; 49.8% of the cows had DD on at least one leg and M4 was the most frequent stage (20.9%). Not removing manure on a regular basis resulted in lower odds for M2, M4 and M4.1 compared to cleaning by automatic scrapers ten times a day or more (odds ratio [OR]= 0.16, 0.49 and 0.18, respectively). The odds for M2 and M4 lesions were higher in cows aged 3-5 years than in first-calved cows (OR> 1.5 and > 1.7, respectively). Rubber flooring in the passageways resulted in lower odds for both M1 and M2 (OR, 0.06 and 0.32, respectively). Prophylactic use of footbaths treatment with an alternative active compound resulted in significant higher odds for M4 lesions than formalin and a combination of formalin and copper sulphate (OR= 1.69 and 2.04 respectively). The odds for an M4.1 lesion were lower in cows from smaller herds (n = 50-100) compared to large herds (n >100; OR= 0.67).

Association between management practices and estimated mastitis incidence and milk losses on robotic dairy farms.

This study aims to describe the relation between farm-level management factors and estimated farm-level mastitis incidence and milk loss traits (MIMLT) at dairy farms with automated milking systems. In this observational study, 43 commercial dairy farms in Belgium and the Netherlands were included and 148 'management and udder health related variables' were obtained during a farm visit through a farm audit and survey. The MIMLT were estimated from milk yield data. Quarter-level milk yield perturbations that were caused by presumable mastitis cases (PMC) were selected based on quarter-level milk yield and electrical conductivity. On average, 57.6 ± 5.4% of the identified milk yield perturbations complied with our criteria. From these PMC, 3 farm-level MIMLT were calculated over a one-year period around the farm visit date: (1) the 'average number of PMC per cow per year', (2) the 'absolute milk loss per cow per day', calculated as the farm-level sum of all milk losses during PMC in one year, divided by the average number of lactating cows and the number of days, and (3) the 'relative milk loss', calculated as the farm-level sum of milk losses during PMC in one year, divided by the estimated total production in the absence of PMC. The 'average number of PMC per cow per year' was on average 1.81 ± 0.47. The PMC caused an average milk loss of 0.77 ± 0.26 kg per lactating cow per day, which corresponded to an average production loss of 2.38 ± 0.82% of the expected production in the absence of PMC. We performed a principal component regression (PCR) analysis to link the 3 MIMLT to the 'management and udder health related variables', whilst reducing the multicollinearity and the number of dimensions. The first principal component was mainly related to 'milking system brand, maintenance and settings'. The second component mainly linked to average productivity and somatic cell counts, whereas the third component mainly contained variables linked with mastitis management, treatment, and biosecurity. The 3 PCR models had R² ranging from 0.46 (for absolute milk loss per cow per day) to 0.57 (for relative milk loss). For all models, the second PC had the largest effect size. This analysis raises awareness of the impact of management factors on a factual basis and provides handles to take management actions to improve udder health.

Hygiene management practices and adenosine triphosphate luminometry of feeding equipment in preweaning calves on dairy farms in Quebec, Canada.

The objective of this study was to describe the cleaning practices currently used for preweaning calves on dairy farms in Quebec, Canada. In addition, contamination of feeding equipment for preweaning calves was described using ATP (expressed as relative light units, RLU), visual assessment, and bacteriological analysis. A questionnaire was administered on 50 commercial dairy farms in Quebec, Canada, regarding the self-reported cleaning protocol used for feeding equipment of preweaning calves. During the visit, a visual score was given to the feeding equipment available at the farm. Afterward, ATP luminometry measurements were obtained using Hygiene UltraSnap and MicroSnap swabs (Hygiene, Camarillo, CA), and the liquid rinsing technique for buckets, nipples, bottles, esophageal tube feeders (ET), the tube of automatic milk feeders (AMF), water samples, and milk replacer. An additional direct swabbing technique was performed on buckets and nipples. The fluid retrieved from the liquid rinsing technique was also used to determine the total bacterial count (TBC) and total coliform count. Based on the bacteriological analysis, optimal RLU cutoff values to determine contamination were obtained. The median (interquartile range) luminometer measurements using the UltraSnap and direct technique for buckets and nipples were 2,082 (348-7,410) and 3,462 (462-7,518) RLU, respectively; and, using the liquid technique for bottles, ET, AMF, water, and milk replacer were 43 (4-974), 15 (4-121), 301 (137-1,323), 190 (71-358), and 94 (38-218) RLU, respectively. Overall, for all equipment and both techniques used, higher RLU values were seen in UltraSnap samples compared with MicroSnap samples. Additionally, for buckets and nipples, higher RLU values were obtained for the direct swabbing method compared with the liquid sampling method for both swabs used. No differences in the level of contamination were seen between the different feeding equipment used within a farm. Overall, a higher correlation with bacteriological results was noticed for ATP luminometry compared with the visual score, with a high correlation for nipples and bottles using the UltraSnap and liquid technique. Based on the classification of "contaminated" (TBC ≥100,000 cfu/mL) or "not contaminated" (TBC <100,000 cfu/mL), optimal ATP luminometer cutoff values for buckets, nipples, bottles, AMF, water, and milk replacer were 798, 388, 469, 282, 1,432, and 93 RLU, respectively. No clear association was found between ATP measurements and the self-reported cleaning protocol. This study gave new insights into the current cleaning procedures and contamination of feeding equipment for preweaning calves on dairy farms in Quebec. In addition, ATP luminometry cutoff values could help benchmark farms regarding cleaning practices and provide customized advice, improving the overall hygiene management, and thus the health, of preweaning calves on dairy farms.

Standardization and validation of ATP luminometry as a diagnostic tool to assess the cleanliness of feeding equipment in preweaning calves.

The objective of this cross-sectional study was to standardize a reliable and repeatable swabbing technique using ATP luminometry (light emission proportional to the amount of ATP with result provided in relative light units [RLU]) to describe the cleanliness of various feeding equipment used for preweaning calves in dairy farms. A total of 7 Québec commercial dairy herds were selected conveniently. Following visual hygiene scoring, the cleanliness of every available piece of feeding equipment was assessed using direct surface swabbing for buckets and nipples with Hygiena UltraSnap swabs. A liquid rinsing technique was used for esophageal feeders, bottles, and automatic milk feeders (AMF) with UltraSnap, AquaSnap, and MicroSnap swabs. To validate direct swabbing technique of buckets, a stage within and between operators was realized, as well as a conventional bacterial culture. A total of 519 swab samples were obtained from 201 pieces of equipment. The median (interquartile range) contamination in RLU for a bottle, esophageal feeder, AMF, bucket and nipple was 2 (1;6), 2 (0;12), 52 (19;269), 886 (128;7,230) and 899 (142;6,928), respectively. The direct swabbing technique, which consists in swabbing directly the surface of an equipment, showed excellent correlation for intrarater reliability (intraclass correlation (ICC) = 0.93; 95% CI: 0.88-0.96). The interoperator (2 sessions with 3 different operators) reliability also showed high correlation (ICC = 0.88; 95% CI: 0.78-0.94 for the first session, and ICC = 0.89; 95% CI: 0.79-0.95 for the second session). Luminometer values were positively associated with the visual score of esophageal feeders, AMF and buckets. A positive correlation between bacterial culture and direct swabbing of buckets was also found for the UltraSnap (rs = 0.653; 95% CI: 0.283-0.873; P = 0.0003) and MicroSnap (rs = 0.569, 95% CI: 0.309-0.765; P = 0.002). This study describes a standardized and practical on-farm swabbing technique for assessing the hygienic status of feeding equipment by luminometry, which can be integrated in the investigation of preweaning dairy calves problems.

Relationship between blood calcium level and post-milking teat canal closure in dairy cows.

The teat canal-one of the primary defense mechanisms of the udder-ensures the milk flow during milking in bovines and prevents pathogens from entering the udder by forming a barrier through the elastic muscle and keratin layers tightly closing the surrounding area. The current study investigated the effects of blood calcium status on teat closure in cows after milking. The study covered 200 healthy teats, of which 100 were from normocalcemic (NC) cows and 100 were from subclinical hypocalcemic (SCH) cows. Teat canal length (TCL) and width (TCW) were measured with ultrasonography at 0-min pre-milking and 15- and 30-min post-milking. Cylindrically shaped teat canal volume (TCV) was calculated by deriving from TCL and TCW. Time-dependent changes in teat canal closure and their relationships with blood calcium levels were analyzed. The results showed that the calcium level did not affect TCL, TCW, and TCV (P > 0.05) during the 15-min post-milking period. However, TCL (P < 0.001), TCW (P < 0.05), and TCV (P < 0.001) were lower in NC cows than in SCH cows at 30-min post-milking. At 15-min post-milking, no correlation existed between the teat canal closure (ΔTCL, ΔTCW, and ΔTCV) and the blood calcium level, while significant correlations were available between the teat canal closure and the blood calcium level {ΔTCL (r: - 0.288, P < 0.001), ΔTCW (r: - 0.260, P < 0.001), ΔTCV (r: - 0.150, P < 0.05)} at 30-min post-milking. The current study concluded that the blood calcium status significantly impacts the teat canal closure in bovines, and calcium status should be meticulously monitored with the mastitis control program to apply necessary strategic steps.

Broad-spectrum infrared thermography for detection of M2 digital dermatitis lesions on hind feet of standing dairy cattle.

Low-effort, reliable diagnostics of digital dermatitis (DD) are needed, especially for lesions warranting treatment, regardless of milking system or hygienic condition of the feet. The primary aim of this study was to test the association of infrared thermography (IRT) from unwashed hind feet with painful M2 lesions under farm conditions, with lesion detection as ultimate goal. Secondary objectives were to determine the association between IRT from washed feet and M2 lesions, and between IRT from unwashed and washed feet and the presence of any DD lesion. A total of 641 hind feet were given an M-score and IRT images of the plantar pastern were captured. Multivariable logistic regression analyses were done with DD status as dependent variable and maximum infrared temperature (IRTmax), lower leg cleanliness score and locomotion score as independent variables, and farm as fixed effect. To further our understanding of IRTmax within DD status, we divided IRTmax into two groups over the median value of IRTmax in the datasets of unwashed and washed feet, respectively, and repeated the multivariable logistic regression analyses. Higher IRTmax from unwashed hind feet were associated with M2 lesions or DD lesions, in comparison with feet without an M2 lesion or without DD, adjusted odds ratio 1.6 (95% CI 1.2-2.2) and 1.1 (95% CI 1.1-1.2), respectively. Washing of the feet resulted in similar associations. Dichotomization of IRTmax substantially enlarged the 95% CI for the association with feet with M2 lesions indicating that the association becomes less reliable. This makes it unlikely that IRTmax alone can be used for automated detection of feet with an M2 lesion. However, IRTmax can have a role in identifying feet at-risk for compromised foot health that need further examination, and could therefore function as a tool aiding in the automated monitoring of foot health on dairy herds.

Mechanism underlying the modulation of milk production by incomplete milking.

Mammary gland secretory activity is modulated by systemic and local factors; however, the relationship between these factors is unknown. The aim of this study was to determine how a local factor, such as incomplete milking, affects mammary epithelial cell activity, number, and responsiveness to blood prolactin (PRL). Eight cows in mid-lactation were differentially milked (i.e., their right quarters were milked incompletely at approximately 70%, and their left quarters were milked completely, twice daily for 4 wk). Throughout the experiment, milk yield was measured at the quarter level. Milk samples were collected from each quarter once a week to assess the milk components, and epithelial cell concentrations, as well as to isolate milk fat globule RNA. In the weeks before and after the experiment, mammary gland functional capacity was evaluated by measuring the volume of milk harvested after complete filling of the gland. At the end of the last experimental week, mammary gland biopsies were performed on each rear quarter. The milk production of quarters milked completely remained stable during the treatment period, whereas, as expected, the milk production of quarters milked incompletely was only 53% of completely milked quarters at the end of the period. Accordingly, the expression of genes related to milk synthesis (CSN2, LALBA, and ACACA) in milk fat was lower in the quarters that were milked incompletely. Incomplete milking decreased the milk lactose content, indicating a loss of integrity of tight junctions. The total yield of epithelial cells in milk was not affected, but their concentration in milk, the BAX:BCL2 gene expression ratio, and the loss of mammary functional capacity were greater in the quarters milked incompletely, suggesting an acceleration of involution in those quarters. The expression of the short isoform of the PRL receptor gene (PRLR) tended to be lower, and the expression of STAT5A and STAT5B tended to decline in the quarters milked incompletely. In mammary gland biopsy samples, the number of both short and long isoforms of the PRLR were not affected, nor were the amount and activation of STAT3 and STAT5. However, the ratio of PRLR short isoform to PRLR long isoform was lower in the quarters milked incompletely. The decrease in milk yield induced by incomplete milking is rapid and associated with a decrease in mammary epithelial cell activity and a decrease in the number of secretory epithelial cells. The results of this experiment provide only limited support for the hypothesis that modulation of the mammary gland's responsiveness to PRL is part of the mechanism by which local factors, such as incomplete milking, modulate milk synthesis.

Assessing optimal frequency for image acquisition in computer vision systems developed to monitor feeding behavior of group-housed Holstein heifers.

Computer vision systems have emerged as a potential tool to monitor the behavior of livestock animals. Such high-throughput systems can generate massive redundant data sets for training and inference, which can lead to higher computational and economic costs. The objectives of this study were (1) to develop a computer vision system to individually monitor detailed feeding behaviors of group-housed dairy heifers, and (2) to determine the optimal frequency of image acquisition to perform inference with minimal effect on feeding behavior prediction quality. Eight Holstein heifers (96 ± 6 d old) were housed in a group and a total of 25,214 images (1 image every second) were acquired using 1 RGB camera. A total of 2,209 images were selected and each animal in the image was labeled with its respective identification (1-8). The label was annotated only on animals that were at the feed bunk (head through the feed rail). From the labeled images, 1,392 were randomly selected to train a deep learning algorithm for object detection with YOLOv3 ("You Only Look Once" version 3) and 154 images were used for validation. An independent data set (testing set = 663 out of the 2,209 images) was used to test the algorithm. The average accuracy for identifying individual animals in the testing set was 96.0%, and for each individual heifer from 1 to 8 the accuracy was 99.2, 99.6, 99.2, 99.6, 99.6, 99.2, 99.4, and 99.6%, respectively. After identifying the animals at the feed bunk, we computed the following feeding behavior parameters: number of visits (NV), mean visit duration (MVD), mean interval between visits (MIBV), and feeding time (FT) for each heifer using a data set composed by 8,883 sequential images (1 image every second) from 4 time points. The coefficient of determination (R2) was 0.39, 0.78, 0.48, and 0.99, and the root mean square error (RMSE) were 12.3 (count), 0.78, 0.63, and 0.31 min for NV, MVD, MIBV, and FT, respectively, considering 1 image every second. When we moved from 1 image per second to 1 image every 5 (MIBV) or 10 (NV, MDV, and FT) s, the R2 observed were 0.55 (NV), 0.74 (MVD), 0.70 (MIBV), and 0.99 (FT); and the RMSE were 2.27 (NV, count), 0.38 min (MVD), 0.22 min (MIBV), and 0.44 min (FT). Our results indicate that computer vision systems can be used to individually identify group-housed Holstein heifers (overall accuracy = 99.4%). Based on individual identification, feeding behavior such as MVD, MIBV, and FT can be monitored with reasonable accuracy and precision. Regardless of the frequency for optimal image acquisition, our results suggested that longer time intervals of image acquisition would reduce data collecting and model inference while maintaining adequate predictive performance. However, we did not find an optimal time interval for all feeding behavior; instead, the optimal frequency of image acquisition is phenotype-specific. Overall, the best R2 and RMSE for NV, MDV, and FT were achieved using 1 image every 10 s, and for MIBV it was achieved using 1 image every 5 s, and in both cases model inference and data storage could be drastically reduced.

A novel approach in the prevention of mastitis: electrical teat dipping.

Teat dipping is widely used in dairy cattle, especially to protect against contagious mastitis. Here we determine the effect of the device called 'Electrical Teat Dipping' (ETD), which was developed by combining teat dipping application and electrical field stimulation technique on teats. For this purpose, the front teats of 100 Holstein breed milking cows were evaluated in two groups, with ETD being applied once to the left front teat of these cows, and conventional teat dipping (CTD) being applied once to the right front teat, both after milking. Ultrasonographic measurements of the teats were made before milking and after teat dipping. We found that the width of the teat canal (1.88 ± 0.07 mm) in the teat using ETD was narrower after the application compared to those with CTD (2.28 ± 0.05 mm). Based on our findings, we conclude that the effects of ETD on the teat are very positive and can potentially be used as a new approach in the preventative control of mastitis in cows.

Comparison of ozonated water and acidified copper sulphate in prevention of digital dermatitis in dairy cows.

Digital dermatitis (DD) is the most significant infectious hoof disorder of cattle in Europe. Hoof baths are one of the most common control methods. Copper sulphate and formalin are commonly used in hoof baths, but their use is problematic in many European countries for health, environmental and safety reasons. Ozonated water and acidified copper sulphate were tested as prevention of DD in a 5-month study. Data were derived from 302 hind feet of Holstein and Estonian Red cows (no. of cows = 151) from a commercial dairy farm in Estonia. Altogether 168 hind feet were included in the acidified copper sulphate group and 134 feet in the ozonated water group. Hoof bathing was carried out three days a week (Mon, Wed, Fri) for two months and then two days a week (Mon, Wed) for three and a half months, in both groups. Ozonated water was sprayed on to the digital skin of hind feet of cows twice a day on treatment days, while the cows were eating. The copper sulphate bath consisted of copper sulphate (2%) mixed with an organic acid compound to acidify and ionize the solution. Cows walked through acidified copper sulphate solution twice a day on treatment days as they were exiting the milking parlor. DD negative and DD positive test results in both groups were compared and statistically tested for differences. The copper sulphate solution was more effective than ozonated water at preventing acute DD lesions. A random maximum likelihood model demonstrated that the odds ratio for DD in the ozonated water group was six times higher compared with DD in the acidified copper sulphate group. Most of the cows that were initially without any DD lesions (M0 + no other severe hoof lesion), remained lesion-free in both groups (copper sulphate group 97% and ozonated water group 88%). Despite trial design deficiencies, the findings indicate that acidified copper sulphate was a more effective solution in preventing DD than ozonated water.

Effect of temporary cessation of milking and estradiol combination on the antimicrobial components in goat milk.

A temporary cessation of milking is widely used in Japan to treat mastitis in dairy cows. Exogenous administration of estradiol (E2) is known to inhibit milk production in dairy cows. Therefore, we aimed to evaluate the effects of the temporary cessation of milking in combination with E2 administration on the antimicrobial components of goat milk. Twelve goats, divided into two groups-with and without E2 injection (E2 and control group, respectively), were subjected to cessation of milking in both udder halves for 3 d (day 0-2). Milk yield in the E2 group was significantly lower than that in the control group on days 7 to 10. The concentrations of cathelicidin-2, IgA, and lactoferrin in the E2 group were significantly higher than those in the control group. These results suggest that the temporary cessation of milking with simultaneous E2 administration leads to a higher concentration of certain antimicrobial components in milk than that observed after using cessation of milking alone. Thus, this combination may contribute to a stronger innate immune system and a faster recovery from mastitis, and might prove to be an alternative to antibiotic treatment upon further research.

Hoof lesions in partly housed pasture-based dairy cows.

Lameness is a symptom of a painful disorder affecting the limbs, which impacts dairy cow welfare and productivity. Lameness is primarily caused by hoof lesions. The prevalence of different lesion types can differ depending on environmental conditions and farm management practices. The aims of this observational study were to establish the cow-level and herd-level lesion prevalence during both housing and grazing periods in a partly housed, pasture-based system, establish the prevalence of lesions always associated with pain ("alarm" lesion), identify the lesions associated with a higher lameness score, determine relationships between lesions, and identify risk factors for digital dermatitis. On 98 farms during the grazing period and on 74 of the same farms during the housing period, every cow was lameness scored (0-3 lameness scoring scale), and the hind hooves of lame cows (score 2 and 3) were examined (maximum 20 cows per visit) and the prevalence of each lesion type recorded. To gather data on potential predictors for the risk factor analysis, a questionnaire with the farmer was conducted on lameness management practices and infrastructure measurements were taken at each visit. Cow-level data were also collected (e.g., parity, breed, milk yield, and so on). Noninfectious lesions were found to be more prevalent than infectious lesions in this system type. The most prevalent lesion types during both grazing and housing periods were white line separation, sole hemorrhages and overgrown claws; all remaining lesions had a cow-level prevalence of less than 15%. The cow-level prevalence of alarm lesions was 19% during the grazing period and 25% during the housing period; the most prevalent alarm lesion was sole ulcers during both periods. We found significantly more foreign bodies within the hoof sole (grazing = 14%, housing = 7%) and overgrown claws (grazing = 71%, housing = 55%) during the grazing period compared with the housing period. Cows with foul of the foot, sole ulcer, white line abscess, toe necrosis or an amputated claw had higher odds of being more severely lame, compared with mildly lame. The strongest correlation between lesions were between toe necrosis and digital dermatitis (r = 0.40), overgrown claws and corkscrew claws (r = 0.33), and interdigital hyperplasia and digital dermatitis (r = 0.31) at herd level. At the cow level, the strongest correlation was between overgrown claws and corkscrew claws (r = 0.27), and digital dermatitis and heel erosion (r = 0.22). The farmers' perception of the presence of digital dermatitis (and lameness) was significantly correlated with the actual presence of digital dermatitis recorded. Additional risk factors for the presence of digital dermatitis were cow track and verge width near the collecting yard, and stone presence on the cow tracks. Results from this study help further our understanding of the causes of lameness in partly housed, pasture-based dairy cows, and can be used to guide prevention and treatment protocols.

Multicow pose estimation based on keypoint extraction.

Automatic estimation of the poses of dairy cows over a long period can provide relevant information regarding their status and well-being in precision farming. Due to appearance similarity, cow pose estimation is challenging. To monitor the health of dairy cows in actual farm environments, a multicow pose estimation algorithm was proposed in this study. First, a monitoring system was established at a dairy cow breeding site, and 175 surveillance videos of 10 different cows were used as raw data to construct object detection and pose estimation data sets. To achieve the detection of multiple cows, the You Only Look Once (YOLO)v4 model based on CSPDarkNet53 was built and fine-tuned to output the bounding box for further pose estimation. On the test set of 400 images including single and multiple cows throughout the whole day, the average precision (AP) reached 94.58%. Second, the keypoint heatmaps and part affinity field (PAF) were extracted to match the keypoints of the same cow based on the real-time multiperson 2D pose detection model. To verify the performance of the algorithm, 200 single-object images and 200 dual-object images with occlusions were tested under different light conditions. The test results showed that the AP of leg keypoints was the highest, reaching 91.6%, regardless of day or night and single cows or double cows. This was followed by the AP values of the back, neck and head, sequentially. The AP of single cow pose estimation was 85% during the day and 78.1% at night, compared to double cows with occlusion, for which the values were 74.3% and 71.6%, respectively. The keypoint detection rate decreased when the occlusion was severe. However, in actual cow breeding sites, cows are seldom strongly occluded. Finally, a pose classification network was built to estimate the three typical poses (standing, walking and lying) of cows based on the extracted cow skeleton in the bounding box, achieving precision of 91.67%, 92.97% and 99.23%, respectively. The results showed that the algorithm proposed in this study exhibited a relatively high detection rate. Therefore, the proposed method can provide a theoretical reference for animal pose estimation in large-scale precision livestock farming.

Differential hepatic mitochondrial function and gluconeogenic gene expression in 2 Holstein strains in a pasture-based system.

The objective of this study was to assess hepatic ATP synthesis in Holstein cows of North American and New Zealand origins and the gluconeogenic pathway, one of the pathways with the highest ATP demands in the ruminant liver. Autumn-calving Holstein cows of New Zealand and North American origins were managed in a pasture-based system with supplementation of concentrate that represented approximately 33% of the predicted dry matter intake during 2017, 2018, and 2019, and hepatic biopsies were taken during mid-lactation at 174 ± 23 days in milk. Cows of both strains produced similar levels of solids-corrected milk, and no differences in body condition score were found. Plasma glucose concentrations were higher for cows of New Zealand versus North American origin. Hepatic mitochondrial function evaluated measuring oxygen consumption rates showed that mitochondrial parameters related to ATP synthesis and maximum respiratory rate were increased for cows of New Zealand compared with North American origin. However, hepatic gene expression of pyruvate carboxylase, phosphoenolpyruvate carboxykinase, and pyruvate dehydrogenase kinase was increased in North American compared with New Zealand cows. These results altogether suggest an increased activity of the tricarboxylic cycle in New Zealand cows, leading to increased ATP synthesis, whereas North American cows pull tricarboxylic cycle intermediates toward gluconeogenesis. The fact that this occurs during mid-lactation could account for the increased persistency of North American cows, especially in a pasture-based system. In addition, we observed an augmented mitochondrial density in New Zealand cows, which could be related to feed efficiency mechanisms. In sum, our results contribute to the elucidation of hepatic molecular mechanisms in dairy cows in production systems with higher inclusion of pastures.

A risk-oriented evaluation of biofilm and other influencing factors on biological quality of drinking water for dairy cows.

Despite the importance of livestock drinking water quality on animal physiology, welfare, and performance, influences such as biofilm formation on trough surfaces on microbial water quality are rarely researched. The objective of this study was to assess the microbial quality of water offered to lactating dairy cows and identify risk factors for poor water quality. We further aimed to determine the impact of biofilm formation on water quality and evaluate rapid test systems to score the hygiene status of dairy troughs on the farm. A total of 105 troughs located on 24 typical Western German dairy farms were sampled. Samples of livestock drinking water and biofilm were analyzed for aerobic total viable count (TVC), coliform count (CC), Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), and other bacteria resistant to 3rd generation cephalosporins (CRB). Surface protein- and adenosine triphosphate (ATP)-rapid tests were evaluated to detect biofilm formation. The influence of 22 selected fixed and variable trough characteristics on impaired livestock drinking water quality was evaluated by calculating odds ratios. The average TVC, CC, and E. coli counts were 4.4 ±â€…0.06 (mean ±â€…SD), 1.7 ±â€…0.1, and 0.6 ±â€…0.1 log10 cfu per mL, respectively. CC was detectable in 94.3% of all water samples and E. coli in 48.6%. MRSA was found in pooled livestock drinking water samples of a single farm and CRB on three farms, suggesting that troughs might function as a reservoir of antibiotic-resistant bacteria, thereby contributing to an exchange of antibiotic-resistant bacteria between animals. Risk factors for the impairment of at least one microbial quality criteria (TVC, CC, or E. coli) increased significantly (P < 0.05) when using high-volume troughs, other trough materials than stainless steel, a lower distance to the milking parlor, heavy visible soiling, biofilm formation, and high ambient and high water temperatures. CC (r = 0.46; P < 0.001) and E. coli (r = 0.31; P < 0.01) of water samples correlated with their equivalent in biofilm and with the results of rapid tests on trough surfaces (0.31 > r > 0.19; P < 0.05). Addressing the identified risk factors could be an approach to ensure sufficient biological quality of livestock drinking water.

Although water is recognized as one of the most important nutrients for dairy cows, only limited research on its quality and potential risk factors for quality impairments is available. This study aimed to evaluate biological water quality on Western German dairy farms, identify potential risk factors for impairment, and evaluate rapid test systems to score the hygienic status of troughs. Water, biofilm, and trough surfaces of 105 water troughs on 24 dairy farms were sampled and analyzed. The bacterial load of sampled livestock drinking water was relatively high. Nearly half of all water samples were contaminated with Escherichia coli, and antibiotic-resistant bacteria were detectable on four farms. Identified risk factors for poor biological water quality that could be used to ensure sufficient water quality on-farm were: high-volume troughs, plastic or cast iron as trough material, a lower distance to the milking parlor, heavy visible soiling, biofilm formation, and high ambient and high water temperatures. Rapid test systems to evaluate trough surfaces correlated with biofilm and water analysis results and could be a useful tool for farmers to check water quality on-farm.

Genetic and phenotypic variation and consistency in cow preference and circadian use of robotic milking units.

Many dairy herds use automatic milking stations (AMS), with cows in large herds often having access to 2 or more AMS, and must choose between them when they go for milking. Individual cows acquire routines of either consistently using a specific milking box or consistently using any available milking box. Here, we hypothesized that the degree of use of the same milking box was an expression of preference, and quantified it as preference consistency score (PCS). The PCS was calculated as a ratio between the excess frequencies of the first choice over the base frequency of "not first choice" over 15-d segments of lactation. This ratio was 0 if all choices were taken equally, and became 1.0 if only the first choice was taken in all events. We investigated the consistency of milking box preference in 2 cohorts (one Holstein and one Jersey) across 6 commercial dairy herds in Denmark (n = 4,665 cows total). In addition to PCS, we recorded and analyzed associated milking and behavior traits, including a time profile index showing use of specific clock hours when cows were milked (Time_profile, based on excess use of specific clock hours), milking frequency, time spent in the milking box, and milk yield. Records from each milking event were condensed into 15-d segments based on days in milk. The data were analyzed using a linear mixed model, with random genetic and individual cow effects, to estimate heritability (h2), repeatability (t), and individual level correlations (ri) between traits. The average PCS was 0.43 and 0.41 in Holstein and Jersey, respectively, showing that cows developed routines for consistently using the same milking box; however, some cows had lower preference (i.e., greater flexibility in use). The Time_profile indicated that some cows were milked in a few hour-bins, whereas others were more flexible. The PCS and Time_profile traits had low heritability (h2, PCS/Time_profile = 0.07 ± 0.02/0.11 ± 0.02 Holstein, 0.13 ± 0.03/0.04 ± 0.02 Jersey) and moderate repeatability (t, PCS/Time_profile = 0.47/0.40 Holstein, 0.50/0.42 Jersey). The 2 traits were weakly correlated with each other (ri = 0.18 and 0.17), and were weakly correlated with milk yield (ri range: 0.0 to -0.10). However, the time profile was strongly correlated with milking frequency (ri range: -0.81 to -0.73), and was moderately correlated with daily box time (ri range: -0.43 to -0.35). In general, Holstein and Jersey parameter estimates were of similar size, and thus in good agreement. Overall, individual cows covered a broad spectrum of preference consistency, both regarding the use of specific milking boxes and time profiles, with these 2 traits representing different aspects or dimensions of milking behavior. The findings that some cows have strong preferences for specific AMS may be most useful in herd management and farm design. The weak correlation to milk yield indicated that yield minimally affected these 2 milking associated behavior traits. In conclusion, although the traits were repeatable, heritability was low; thus, genetic selection for milk yield might minimally affect these 2 traits.

Alberta dairy farmers' and veterinarians' opinion about bovine leukemia virus control measures.

A high herd and within-herd prevalence of Bovine Leukemia Virus (BLV) infections in the dairy herds of North America and the negative effects thereof caused the Alberta dairy industry to initiate the development of an on farm BLV control program. Because BLV control is dependent on the commitment of the farmer, potential barriers were identified and farmers' and veterinarians' points of view toward different control options were investigated to inform how the control program might be adjusted. Conversations with these stakeholders were sought and four focus groups with farmers and eleven interviews with veterinarians were conducted. Testing for BLV, the most common BLV control strategies (testing/culling/segregation/management), as well as on farm best management practices (BMP) to prevent the transmission of BLV, were discussed. The thematic analysis of these conversations resulted in the following findings: Testing of animals was considered important for BLV control, but the financial investment was prohibitive for farmers. Test and cull as well as test and segregation approaches of test positive animals were considered efficient BLV control measures, but impractical and not feasible due to the supply managed Alberta dairy industry (i.e. milk is produced based on demand), with a high prevalence. The management of test positive animals with BMP to prevent new infections and thereby decreasing the within-herd prevalence was considered the only realistic BLV control strategy. The most important barriers for suggested BMP were the cost for some BMP, the inconvenience of performing other BMP, as well as difficulties in performing some BMP consistently and well. Additionally, a lack of knowledge about BLV and its control were identified as an important barrier. On the contrary, farmers indicated being inclined to implement BMP they considered feasible or that were considered a standard within the industry. Further, if BMP increased convenience on farm, they were considered easy to implement. Farmers and veterinarians agreed in many, but not all cases. For example, the single use of examination sleeves was met with differing opinions (i.e. considered doable by farmers while veterinarians assumed it to be too costly). In conclusion, stakeholders' awareness and communication amongst each other (e.g. veterinarians and farmers) about BLV and its control has to be highlighted in order to manage BLV infection successfully. In addition, by communicating and understanding barriers and motivators for specific BMP, important barriers could be identified (e.g. difficulties while changing needles), and solutions found (e.g. tool belt for needles), thereby improving BLV control efforts on farm.

A reliable method using the cytobrush for diagnosis of subclinical endometritis in dairy cattle during the late lactational period.

This experiment was performed to assess reliability of the cytobrush-cytology method (CCM) in diagnosis of subclinical endometritis (SCE) using the biopsy-histopathology method (BHM) as a reference in late lactating dairy cows. Reproductive organs were collected from 115 slaughtered multiparous crossbred cows culled due to infertility 398 ± 135 days subsequent to parturition. Samples were collected from the dorsal part of the corpus uteri for analyses. Inflammation status was graded histopathologically based on the cell percentages [(neutrophils, eosinophils, lymphocytes (LYM), macrophages (MAC), and plasma cells)]. Data were subjected to Friedman's test for group comparisons (method and diagnosis), concordance correlation and chi-square tests for consistency of results among methods, and the receiver operating characteristics curve analysis for reliability of the CCM. Percentages of LYM (2.67x) and MAC (3.00x) were greater when evaluated using BHM than with CCM (P < 0.05 for both). The agreement (Cohen's κ value) of results among methods was 0.79 ± 0.06. The sensitivity (Se) and specificity (Sp) of the CCM for defining endometrial inflammation were 79.3% and 100%, respectively. Among inflammatory cells, proportions of LYM and MAC in the CCM had merit for evaluation of uterine inflammation, with an Se of 74.1 and 84.5 and an Sp of 93.0 and 75.4 at the cut-off > 4 and > 0, respectively. The results indicate the CCM may be used in the diagnosis of SCE when the LYM and MAC percentages are considered in chronically infertile cows in the later stages of the lactational period.

Isolation and Identification of Lactic Acid Bacteria from Cow Milk and Milk Products.

Lactic acid bacteria (LAB) have long been consumed by people in several fermented foods such as dairy products. A study was conducted on lactating dairy cows to isolate and characterize LAB from dairy products found in and around Bahir-Dar city, North Western Ethiopia. Milk and milk products were randomly collected from dairy farms, milk vending shops, individual households, and supermarkets for bacteriological investigations. A total of sixteen samples were taken from different sources and cultured on different selective media: de Man, Rogosa, and Sharpe (MRS) agar for Lactobacillus spp.; M17 agar for Lactococcus spp.; Rogasa SL agar for Streptococci spp.; and MRS supplemented with cysteine (0.5%) for Bifidobacteria spp. Different laboratory techniques were implemented for LAB isolation and identification. A total of 41 bacterial isolates were grouped under five different genera of LAB and Bifidobacteria spp. were identified based on the growth morphology on the selective media, growth at a different temperature, gas production from glucose, carbohydrate fermentation, and other biochemical tests. LAB genera such as Lactobacillus, Lactococcus, Leuconostoc, Pediococcus, Streptococcus, and Bifidobacterium spp. were isolated and identified from raw milk, cheese, and yogurt. Based on the current study, the majority of the LAB (24.38%) was isolated from cheese and yogurt. Among these, Lactobacillus, Lactococcus (21.94%), Streptococcus (19.51%), Leuconostoc (14.64%), Bifidobacteria (12.19%), and Pediococcus (7.31%) spp. were also identified from these products. Furthermore, based on the bacterial load count and different identification methodologies, our study revealed that Lactobacillus spp. were the dominant LAB isolated from milk and milk products. As a result, since there are few studies on the isolation and identification of lactic acid bacteria from dairy products in Ethiopia, more research studies are needed to complete the identification and characterization to species level and their possible role as probiotics.

Effect of exogenous administration of oxytocin on postpartum follicular dynamics, oestrous rate and ovulation in Nili-Ravi buffaloes.

Based on different surveys, dairy farmers are concerned about extensive use of exogenous oxytocin in buffaloes, which is being held responsible for reproductive problems including irregular oestrous cycle and delayed ovulation. For these concerns, effects of oxytocin injection on postpartum follicular dynamics, postpartum oestrous interval (PEI), oestrous length, the interval from onset of estrus to ovulation and blood progesterone (P4) were studied in Nili-Ravi buffaloes. For this purpose, 23 animals within 1 week after calving were randomly divided into three groups: without oxytocin (CON; n = 7), 10 i.u. oxytocin (LOW; n = 8), 30 i.u. oxytocin - (HIGH; n = 8) and used to record the PEI for the study period of 154 days. At subsequent estrus, three buffaloes from each group (not served) were selected randomly to monitor two cycles for 6 weeks. Transrectal ultrasonography was performed to evaluate follicular and corpus luteum (CL) development, and blood sampling was done for progesterone (P4) analysis. These results revealed that postpartum oestrous interval (PEI) decreased significantly in oxytocin-treated groups. The number of small, medium and total follicles on the left ovary was significantly higher in the HIGH group. However, an overall number of small and total follicles on both right and left ovaries was significantly higher in CON and HIGH groups. On the other hand, there was no difference in the number of follicles on the right ovary among all treatment groups. The same was true for the size of pre-ovulatory follicles, CL, P4 concentrations and oestrous cycle length. The intervals from onset of estrus to ovulation and from standing estrus to ovulation were increased considerably in the HIGH group. It is concluded that exogenous oxytocin administration resulted in the shortening of PEI but triggered a delay in ovulation. Moreover, a higher dose of oxytocin could stimulate the growth of small, medium, and total follicles in postpartum Nili-Ravi buffaloes.