Accuracy of direct and indirect methods for assessing bovine colostrum quality using a latent class model fit within a Bayesian framework.

Feeding high-quality colostrum is essential for calf health and future productivity. Therefore, accurate assessment of colostrum quality is a key component of dairy farm management plans. Direct and indirect methods are available for assessment of colostrum quality; however, the indirect methods are rapid, inexpensive, and can be performed under field settings. A hierarchical latent class model fit within a Bayesian framework was used to estimate the sensitivity (Se) and specificity (Sp) of the radial immunodiffusion (RID) assay, transmission infrared (TIR) spectroscopy, and digital Brix refractometer for the assessment of low-quality bovine colostrum in Atlantic Canada dairy herds. The secondary objective of the study was to describe the distribution of herd prevalence of low-quality colostrum. Colostrum quality of 591 samples from 42 commercial Holstein dairy herds in 4 Atlantic Canada provinces was assessed using RID, TIR spectroscopy, and digital Brix refractometer. The accuracy of all tests at different Brix value thresholds was estimated using Bayesian latent class models to obtain posterior estimates [medians and 95% Bayesian credibility intervals (95% BCI)] for each parameter. Using a threshold of <23% for digital Brix refractometer and <50 g/L for RID and TIR spectroscopy, median (95% BCI) Se estimates were 73.2 (68.4-77.7), 86.2 (80.6-91.0), and 91.9% (89.0-94.2), respectively. Median (95% BCI) Sp estimates were 85.2% (81.0-88.9) for digital Brix refractometer, 99.4% (97.0-100) for RID, and 90.7% (87.8-93.2) for TIR spectroscopy. Median (95% BCI) within-herd low-quality colostrum prevalence was estimated at 32.5% (27.9-37.4). In conclusion, using digital Brix refractometer at a Brix threshold of <23% could reduce feeding of low-quality colostrum to calves and improve colostrum and calf management practices in Atlantic Canada dairy herds. The TIR spectroscopy showed high Se in detection of low-quality colostrum. However, the RID assay, which is used as the reference test in several studies, showed limited Se for detection of low-quality colostrum.

Relationships between type of hoof lesion and behavioral signs of lameness in Holstein cows housed in Canadian tiestall facilities.

Although foot pain can affect gait, the presence of a hoof lesion may or may not cause the cow to show visible changes in their gait. This can be dependent on the type and severity of the lesion; for example, the presence of a sole ulcer (SU) has been associated with increased gait scores, whereas digital dermatitis (DD) and sole hemorrhage (SH) have not. In tiestall facilities, gait scoring can be difficult to perform. An alternative method, known as stall lameness scoring (SLS), allows observers to assess cattle for lameness while they remain in their stall. Lameness is determined based on behavioral changes in weight bearing and foot positioning, which include: shifting weight, resting a foot, standing on the edge of the stall, and uneven weight bearing when stepping side to side. The aim of this study was to examine relationships between hoof lesions and these behavioral indicators. A total of 557 observations of SLS and corresponding hoof trimming records, collected during routine trimming events on 7 tiestall herds, were obtained. Trimming was performed by 2 trained hoof trimmers with good agreement on lesion identification, based on quizzes taken at the beginning and mid-way through the study. To ensure trimming had no effect on the behavioral indicators observed, SLS was always performed by a trained observer before trimming. Behavioral indicators focused on the hind limbs only; therefore, the analysis was confined to hind limb lesions using logistic regression to detect the presence of hoof lesion based on observations made during SLS. Seventy-five percent of observed cows had no SLS behavioral indicators, whereas, 11, 12, and 1% had 1, 2, and 3 behavioral indicators, respectively. At least one hind limb lesion was noted during trimming in 19% of cows, with the most common lesions being DD (7%), SU (6%), and SH (4%). A cow that was observed resting one foot and bearing weight unevenly when moving side to side had higher odds of having a hind limb hoof lesion than a cow not displaying these behaviors. When looking at specific hoof lesions, a cow observed resting one limb and bearing weight unevenly had higher odds of having a SU compared with those not displaying these behaviors. A cow observed shifting their weight from one foot to another had higher odds of having SH, and a cow observed bearing weight unevenly had higher odds of DD. Behavioral indicators in weight bearing and foot positioning can help identify cows in tiestalls with hind limb hoof lesions. Producers could routinely observe their cattle for these indicators to assist in the identification of cows that may require treatment. This could help reduce the duration of clinical lameness through earlier intervention.

Associations of freestall design and cleanliness with cow lying behavior, hygiene, lameness, and risk of high somatic cell count.

The objective of this study was to investigate associations of freestall design and cleanliness with cow lying behavior, hygiene, lameness, and risk of new high somatic cell count (SCC). Cows from 18 commercial freestall dairy herds (22 ± 15 cows/farm; mean ± SD) in Ontario, Canada, were enrolled in a longitudinal study. Four hundred focal cows that were <120 d in milk, had no mastitis treatment in the last 3 mo, and had an SCC <100,000 cells/mL at their most recent milk test were selected for the study. Data on SCC were collected through Dairy Herd Improvement Association milk testing (at ~5-wk intervals). Each farm was visited 5 ± 3 d (mean ± SD) after each milk test until 3 tests were completed (~105 d), for a total of 3 observation periods per cow. Elevated SCC was used as an indicator of subclinical mastitis. An incident of new high SCC was defined as a cow having SCC >200,000 cells/mL at the end of an observation period, when SCC was <100,000 cells/mL at the beginning of that period. Lying behavior was recorded for 6 d after each milk sampling, using electronic data loggers. Cows were scored during each period for lameness (5-point scale, with scores ≥3 = lame), body condition score (BCS; 5-point scale; 1 = thin to 5 = fat), and hygiene (4-point scale). Stall cleanliness was assessed during each period with a 1.20 × 1.65-m metal grid, containing 88 squares. The grid was centered between stall partitions of every tenth stall on each farm, and the squares containing visible urine or fecal matter (or both) were counted. Cow lying time averaged 10.9 ± 1.9 h/d. On average, cows with low BCS (≤2.5) spent 37 ± 16.6 min/d less time lying down than high-BCS cows (≥4.0). On average, cows tended to spend 36 ± 18.3 min/d more time lying down in deep-bedded versus mattress-based stalls. Mean proportion of soiled squares per stall was 20.1 ± 0.50%. Across farms, cow lying time decreased as the proportion of soiled squares per stall increased. A difference in daily lying time of ~80 more min/d was modeled for cows housed in barns with the cleanest stalls compared with those with the dirtiest stalls. Higher neck rail height [for every 1 SD (10 cm) increase] increased the odds (odds ratio = 1.5) of cows having a dirty upper leg-flank and udder. The odds of a cow having a dirty upper leg-flank, udder, and lower legs were 1.5, 2.0, and 1.9 times greater, respectively, for cows housed with dirtier stalls. Also, cows housed on farms with dirtier stalls had 1.3 times greater odds of being lame at the time of observation. Over the study period, 50 new high-SCC cases were detected, resulting in an incidence rate of 0.45 cases of new high SCC per cow-year at risk. No measured factors were detected to be associated with risk of a new high SCC. Overall, our results confirm that cows lie down longer in cleaner and more comfortable environments. Further, these results highlight the need for improved stall cleanliness to optimize lying time and potentially reduce lameness.

Communication preferences and social media engagement among Canadian dairy producers.

The objective of this study was to determine communication preferences of dairy producers in Canada. A secondary objective was to evaluate social media engagement of dairy producers. A survey was administered to Canadian dairy producers between March and April, 2015 to collect information on current management practices on their farms. A total of 1,373 Canadian dairy producers responded to the survey, representing a response rate of 12%. The survey consisted of 192 questions; however, only questions regarding producer demographics, importance of information sources, and internet and social media use were evaluated in this study. The primary outcome variables of interest included use of the internet to access dairy information, importance of different sources of information about dairy herd health and management, and use of online search engines and social media platforms. For each outcome, logistic regression analyses were used to investigate associations between the outcome and demographic variables. Veterinarians were viewed as a "very important" source of information by the majority of respondents (79%), whereas milk recording and dairy producer organizations were viewed as a "very important" source of information by 36% of respondents. Other producers (46%) and magazines or newspapers (51%) were commonly viewed as an "important" source of information. Online search engines were commonly used by respondents (94%). Social media was viewed as less important, and had mixed levels of use. YouTube (70%), Facebook (63%), and Twitter (18%) were the most commonly used social media platforms. Eighty percent of Twitter users reported using the platform to interact with and obtain or share information about herd health management online, which was the highest reported interactivy regarding herd health among all social media platforms. This exploratory study offers insight into the communication preferences of Canadian dairy producers and can be used to facilitate future communication strategies aimed at engaging rural farming audiences across Canada.

Short communication: Describing mortality and euthanasia practices on Canadian dairy farms.

The objective of this study was to describe mortality rates and euthanasia practices used for cows, heifer calves, and male dairy calves on Canadian dairy farms. An internet survey was administered to Canadian dairy producers between March and April 2015 to collect information on current management practices. Approximately 81% (867/1,076) and 63% (673/1,065) of respondents reported that at least one animal died unassisted and at least one animal was euthanized on the farm in the preceding 12-mo period, respectively. Overall, mean mortality was 8% for cows, and 6 and 2% for preweaning and weaned heifers, respectively. On average, 48, 76, and 89% of all reported mortality events in cows, weaned heifers, and preweaning heifers were recorded as unassisted deaths. Cows that died without assistance were necropsied more often than preweaning heifers dying without assistance; these cows were also necropsied more than cows that were euthanized. Conversely, preweaning heifers that were euthanized were necropsied more frequently than those that died without assistance. Choosing not to perform necropsies on animals that die of unknown causes could represent a missed opportunity to identify cause of death and inform changes to prevent future deaths. The use of a firearm was the most common method for euthanizing cows and heifers (54%) and male dairy calves (51%). Approximately 7% of respondents reported using blunt force to euthanize animals. The use of this method was much higher for euthanasia of male dairy calves (34%) than for heifer calves (7%). Québec respondents euthanized their animals using blunt force more often than farmers from other regions. Further, 53% of respondents using blunt force indicated it was their primary method of calf euthanasia; these results are extremely concerning. Approximately 31% of respondents who reported that they do not use blunt force to euthanize heifers and cows did report using blunt force to euthanize male dairy calves. These results can be used to inform veterinarian-client communication, broader extension tools and programs, and industry policies to improve dairy cattle health and welfare, a goal that is becoming of increasing importance from a consumer assurance perspective.

Canadian National Dairy Study: Describing Canadian dairy producer practices and perceptions surrounding cull cow management.

Farmer decisions surrounding culling have an important effect on the health and welfare of cull cows. The objectives of this study were to describe the self-reported shipment behaviors of Canadian dairy producers and understand farmer perspectives on the factors that were most influential in their decision to cull a cow. A nationwide survey was administered between March and April 2015 that included 192 questions covering producer background information, farm characteristics, biosecurity practices, disease prevalence, calf health, cow welfare, lameness, milking hygiene, reproduction, and Internet and social media use. The survey yielded a 12% response rate; a total of 1,076 respondents (78% of all survey respondents completed the culling section of the survey) were included in this study for analysis. Approximately 80, 51, and 38% of respondents reported shipping at least 1 cow to auction, direct to slaughter, and to another dairy farm in the past 12 mo, respectively. Ability of the cow to remain standing (93% of respondents) and drug withdrawal times (92% of respondents) were identified as the most important factors for consideration when culling cows. The time between culling decision and when the cow was actually transported was longer for lame cows than sick cows; almost 70% of respondents reported that cows culled for illness were typically shipped within 1 wk of culling decision, whereas only 51% of respondents indicated the same was true for lame cows. Last, Canadian dairy producers generally exhibited strong confidence that their culled cows would arrive at slaughter in the same condition as they left, but felt very unsure about knowing the location of their final destination. These results highlight several gaps between producer perceptions and the true situation, and can be used to develop tailored programs and inform policy and regulatory decisions aimed at improving cull cow decisions and cow welfare.

Canadian dairy farmers' perception of the efficacy of biosecurity practices.

The adoption of biosecurity practices on dairy farms is limited worldwide. Multiple aspects, one of which is the perception of the effectiveness of these practices, influence the adoption of preventive behavior. The objectives of the present study were to investigate the general understanding of biosecurity by Canadian dairy farmers and to describe their perception of the effectiveness of specific biosecurity practices. In 2015, 368 Canadian dairy farmers were selected to participate in an on-farm national cross-sectional benchmarking study during which they were asked about their general understanding of biosecurity and their perception of the effectiveness of 17 practices to minimize biosecurity risk. For these same biosecurity practices, respondents were asked whether they discussed that particular practice with a veterinarian and whether they implemented it on their farm. Herd and respondent characteristics were also recorded. Associations between the perception of effectiveness of each biosecurity practice and the discussion with a veterinarian, its implementation on farm, and the herd and respondent characteristics were examined using ordered regression models. Most respondents thought the purpose of on-farm biosecurity was to prevent both entry of a new pathogen and spread of an existing pathogen (73%) and considered general biosecurity to be effective (92%) and important (58%). When asked about specific biosecurity practices, respondents considered most of them to be effective (60-94%). Practices related to direct animal-to-animal contact were perceived as effective by more respondents than biosecurity practices related to fomites and visitors. Less than 20% of the respondents reported discussing the different biosecurity practices with a veterinarian, and less than 60% reported implementing these practices on their farm. Geographical region, involvement of a veterinarian, and implementation of the practice on farm were associated with the perception of effectiveness for multiple practices. Finally, the reasons chosen by most respondents as motivation to implement biosecurity practices were (1) if the practice helped to regionally prevent the introduction or the spread of a disease and (2) if it was proven to be beneficial to the health and welfare of animals. These results show that there is a positive perception of the effectiveness of specific biosecurity practices, that there might be a lack of understanding of the practices involving transmission of diseases via indirect contact, and that the perceived threat associated with not adopting practices is minimal.

Biosecurity and herd health management practices on Canadian dairy farms.

The Canadian dairy industry has created national standards to support the adoption of biosecurity practices and to reduce disease risks across the country. There is, however, very little information on the adoption of these practices. The present study aimed to describe the adoption of biosecurity practices on Canadian dairy farms shortly after the creation of the national standards and to identify demographic factors associated with practice adoption. In 2015, 2 questionnaires (phase 1 and 2) were administered to Canadian dairy farmers during an extensive cross-sectional study. Associations between adoption of biosecurity practices as well as associations between adoption of these practices and demographic variables were tested using multiple correspondence analysis. A total of 1,157 questionnaires were completed in phase 1, and a subsample of 368 respondents was selected using stratified random sampling to complete phase 2 during visits to the farms. There was a lack of investigation into general disease syndromes such as a high prevalence of abortion or an unexplained death (38 and 22% of respondents, respectively). Biosecurity measures within herds and between herds to minimize the spread of infection were not widely adopted (e.g., 27% of the respondents never housed sick or lame animals in their calving pen, 41% had closed herds, and 25 and 48% of the open herds had no strategy for introducing new additions and reintroducing returning animals, respectively). Cleanliness of the cows before calving was always ensured by 29% of the respondents, and 27% of the respondents reported always sanitizing the calving pen after each calving. Less than 15% of the respondents had measures in place to limit or control visitors coming on their farm. Moreover, less than half of the respondents reported requiring family members, employees, and visitors to wear farm-designated or clean boots and coveralls. From the multiple correspondence analysis, 2 dimensions were retained and were summarized as "animal movement, calving area, and visitor biosecurity" and "employee biosecurity." Geographical region, type of housing, and milk production were associated with the "employee biosecurity" dimension. The present study demonstrates that many important biosecurity practices are not implemented on Canadian dairy farms; therefore, efforts to promote the adoption of these practices will be necessary to improve biosecurity in dairy herds.

Using serum and plasma samples to assess failure of transfer of passive immunity in dairy calves.

The objectives of this study were (1) to determine the differences in IgG and total protein (TP) content of serum and plasma samples collected from the same calves; (2) to evaluate the correlation between calf serum and plasma IgG levels, Brix scores, and TP concentrations; (3) to determine whether different cut-off values should be used for plasma and serum to assess failure of transfer of passive immunity (FTPI) in dairy calves; and (4) to evaluate the level of agreement between results obtained from using serum and plasma samples of the same calves to assess FTPI using optimal cut-off values. Blood samples (n = 217) were collected from Holstein calves at 3 to 10 d of age on 30 commercial dairy farms in Nova Scotia and Newfoundland, Canada. Paired serum and plasma samples were analyzed for IgG concentration by the reference radial immunodiffusion assay, transmission infrared (TIR) spectroscopy, digital and optical Brix refractometers, and optical TP refractometer. The IgG concentrations measured by RID and TIR spectroscopy in serum were similar to those in plasma. However, the Brix and TP refractometer readings were significantly higher in plasma than in serum. The prevalence of FTPI in serum and plasma samples based on a RID-IgG concentration <10 g/L was 43.3 and 46.5%, respectively. The RID-IgG concentration was correlated with TIR-IgG (r = 0.92 and 0.89), digital Brix (r = 0.80 and 0.80), optical Brix (r = 0.77 and 0.77), and optical TP (r = 0.75 and 0.77) refractometers in serum and plasma, respectively. The correlations between paired serum and plasma IgG content were 0.85 by TIR spectroscopy, 0.80 by digital Brix, 0.77 by optical Brix, and 0.79 by optical TP refractometer. The optimal cut-off values for TIR spectroscopy, digital Brix, optical Brix, and TP refractometers to assess FTPI using serum were 13.1 g/L, 8.7% Brix, 8.4% Brix and 5.1 g/dL, respectively; and the optimal cut-off values with plasma were 13.4 g/L, 9.4% Brix, 9.3% Brix and 5.8 g/dL, respectively. When using these optimal cut-off values, the level of agreement (88.1%) between results derived from testing serum and plasma by TIR spectroscopy was substantial, with a kappa (κ) value of 0.76. The results derived from testing serum and plasma by digital Brix refractometer showed substantial agreement (83.4%), with a κ value of 0.65, which is higher than the agreement and κ value (74.7% and 0.51) reported for the optical Brix refractometer. Substantial agreement (81.6%) between serum and plasma TP was also obtained when using the optical TP refractometer, with a κ value of 0.63. In conclusion, serum or plasma samples can be used interchangeably for measuring IgG concentrations and assessing FTPI in dairy calves. However, different cut-offs must be used to assess FTPI depending on the sample matrix. Furthermore, results obtained from serum samples showed higher agreement with the reference RID assay than those obtained from plasma samples.

Prevalence of lameness and associated risk factors on dairy farms in the Maritime Provinces of Canada.

Lameness in dairy cattle is a major issue for the industry due to the effects on the welfare of the animal, the economic impact, and consumer perception. The aim of this study was to determine the prevalence of lameness and explore potential risk factors in the Maritime Provinces of Canada. Cows were scored for lameness and potential risk factors and were assessed in 46 freestall herds and 33 tiestall herds in Nova Scotia, New Brunswick, and Prince Edward Island. In freestall herds, lameness was assessed using the most common method, locomotion scoring. A cow with a gait score of ≥3 out of 5 was considered to be lame. In tiestall herds, lameness was assessed using an alternative method known as stall lameness scoring. This assessment consisted of observation of the cow for 4 behavioral changes: standing on the edge of the stall, shifting weight, resting a limb, and uneven weight bearing when moved side to side. A cow displaying 2 or more of these behaviors was considered to be lame. At the time of the assessment, other animal-, environmental-, and management-based measurements were collected. These measurements were used in multivariable logistic regression analysis to determine risk factors that were associated with lameness for both freestalls and tiestalls independently. The prevalence of lameness was 21% for freestall-housed cattle and 15% for tiestall-housed cattle. Of the 1,488 tiestall-housed cows that were assessed, 68% showed no behavioral changes, whereas 15, 15, 2, and <1% showed 1, 2, 3, or 4 changes, respectively. In freestalls, higher odds of lameness were seen when cows spent ≥3 h/d in the holding area for milking compared with those that spent <3 h/d. In tiestall herds, higher odds of lameness were seen when bedding material was wet compared with when it was dry. For both lactating cow facility types, housing the dry cows and heifers on a deep bedded pack compared with tiestalls or freestalls was associated with a decreased odds of lameness. There were also many cow-level variables associated with lameness, including parity, daily milk production, stage of production, body condition, and width at the tuber coxae (hook bones). If producers become aware of the risk factors associated with lameness, they can make informed decisions on where to implement changes to help reduce the level of lameness in their herd.

Prevalence of hock, knee, and neck skin lesions and associated risk factors in dairy herds in the Maritime Provinces of Canada.

Skin lesions are commonly seen in dairy herds and have been associated with animal-, environmental-, and management factors. These lesions are not only a welfare concern, but they also affect profitability. Three areas on the cattle were examined for skin lesions: the hock, knee, and neck. Previous Canadian studies estimating the prevalence of lesions and the risk factors associated with them have not included the Maritime Provinces. In this study, 73 herds in the Maritime Provinces were chosen voluntarily to participate, with both tiestalls (n = 33) and freestalls (n = 40) represented. Within each herd, 67 to 90% of the lactating cows were selected and assessed for potential animal-, environmental-, and management-based risk factors. If producers were aware of the potential risk factors, this could help them reduce the prevalence in their herd. Leg lesions were scored on a 4-point scale (0-3) based on hair loss, swelling, and scabs, with a lesion defined as a score of 2 or 3 on at least 1 hock or knee. Necks were scored on a 3-point scale (0-2), with a lesion defined as score 2. For freestalls, the prevalence (95% confidence interval) of hock lesions was 39% (29-49%), knee lesions was 14% (11-18%), and neck lesions was 1% (<1-2%). Similarly, for tiestalls the prevalence (95% confidence interval) of hock lesions was 39% (33-46%), knee lesions was 17% (13-22%), and neck lesions was 5% (3-8%). Due to differences in management and methods of assessment between facility types, tiestalls and freestalls were analyzed separately. Due to dichotomization of cows as having a skin lesion or not, random-effects multivariable logistic regression was used to determine the risk factors for each lesion and facility type. Several environmental-based measurements, such as the stall base, type and dryness of bedding, and type of milking parlor, were associated with leg lesions. An environmental-based measurement that was associated with neck lesions was the design of the feed rail barrier in freestalls and the dimensions of the tie rail in tiestalls. Animal-based risk factors, such as stage of lactation, parity, and body condition, were also associated with all 3 types of lesions. This study showed that lesions to the hock, knee, and neck were common in the Maritime Provinces of Canada. Although differences were seen between facility types, in general, the results suggest that improving stall design and management and feed bunk design would help producers reduce the number of skin lesions seen in dairy cattle.

Canadian National Dairy Study: Herd-level milk quality.

The objective of this study was to estimate Canadian national milk quality parameters and estimate the bulk tank milk (BTM) prevalence of 4 mastitis pathogens, Staphylococcus aureus, Streptococcus agalactiae, Mycoplasma bovis, and Prototheca spp., on Canadian dairy farms. A questionnaire was sent to all Canadian dairy producers. Of the 1,062 producers who completed the questionnaire, 374 producers from across the country were visited and milking hygiene was assessed. Farm-level milk quality data for all Canadian dairy producers was collected from the provincial marketing boards and combined with the questionnaire and farm visit data. In addition, a BTM sample was collected either during the farm visit or by the marketing board in November of 2015 and was tested for 4 major mastitis pathogens using the PathoProof Mastitis Major 4 PCR Assay (Thermo Fisher Scientific Inc., Waltham, MA). Apparent herd-level prevalence was 46% for S. aureus, 6% for Prototheca spp., 0% for M. bovis, and 0% for Strep. agalactiae. Due to the low prevalence of M. bovis and Strep. agalactiae and a lack of significant factors associated with farms testing positive for Prototheca spp., an association analysis could only be carried out for Staph. aureus-positive farms. Factors associated with Staph. aureus-positive farms were not fore-stripping cows before milking (odds ratio = 1.87), milking with a pipeline system (odds ratio = 2.21), and stall bases made of a rubberized surface (mats and mattresses), whereas protective factors were using blanket dry cow therapy (odds ratio = 0.49) and applying a tag or visible mark on cows known to have chronic mastitis infections (odds ratio = 0.45). The Canadian national production-weighted geometric mean somatic cell count was determined to be 208,000 cells/mL. This is the first national dairy study conducted in Canada. Participating farms had higher milk yield; were more likely to have a loose housing system, parlor, or automated milking system; and had lower weighted mean BTM somatic cell count than the national level. Sampling larger farms with better milk quality means the apparent prevalence of the 4 mastitis pathogens likely underestimates the true levels.

Short communication: Evaluation of MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci.

This study evaluated MALDI-TOF mass spectrometry and a custom reference spectra expanded database for the identification of bovine-associated coagulase-negative staphylococci (CNS). A total of 861 CNS isolates were used in the study, covering 21 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 804) and the remainder were identified by sequencing of hsp60 (n = 56) and tuf (n = 1). The genotypic identification was considered the gold standard identification. Using a direct transfer protocol and the existing commercial database, MALDI-TOF mass spectrometry showed a typeability of 96.5% (831/861) and an accuracy of 99.2% (824/831). Using a custom reference spectra expanded database, which included an additional 13 in-house created reference spectra, isolates were identified by MALDI-TOF mass spectrometry with 99.2% (854/861) typeability and 99.4% (849/854) accuracy. Overall, MALDI-TOF mass spectrometry using the direct transfer method was shown to be a highly reliable tool for the identification of bovine-associated CNS.

Comparison of fecal pooling strategies for detection of Mycobacterium avium ssp. paratuberculosis in cattle.

In herds with typical moderate to low within-herd prevalence, testing for Mycobacterium avium ssp. paratuberculosis (MAP), the infectious agent of Johne's disease, will be more cost-effective if individual fecal samples are cultured in composite pools. However, sensitivity to classify a pool containing 1 or more positive individual samples as positive may depend on pool size and number of individual positive samples within a pool. Fecal samples collected from 994 dairy cows sampled at slaughter were cultured to detect MAP. Culturing was done both individually and as composite pooled samples using the TREK ESP Culture System II broth medium (Thermo Fisher Scientific, Trek Diagnostic Systems Inc., Cleveland, OH). Composite samples consisted of pools containing feces from 3, 5, 8, 10, or 15 cows. The number of individual fecal culture-positive cows within each pool ranged from 0 to 4. Culture of individual fecal samples detected MAP in 36 (3.6%) of the 994 cows. Individual samples that were detected within the first 50 d by TREK ESP Culture System II were more likely to lead to a positive pool result. In total, 840 pooled fecal samples were examined for presence of MAP, and of those, 272 pools actually contained feces from fecal culture-positive cows. The crude sensitivity (proportion of pools that contained at least 1 fecal-positive cow that tested positive) for pools of 3, 5, 8, 10, and 15 was 47, 67, 44, 59, and 39%, respectively. Across pools, an increase of the number of fecal culture-positive samples from 1 to 2 enhanced overall crude sensitivity from 44 to 71%. However, sensitivity did not further increase for pools with 3 or 4 fecal culture-positive samples (63 and 60%, respectively). Additionally, a simulation analysis assessing probability of pooled fecal samples being positive in herds of 50 and 100 cows was conducted. The simulation assumed that 1, 2, or 5 cows per herd were MAP fecal culture-positive and that pools of 5 and 10 were used. This low-prevalence herd simulation indicated that weighted mean herd probabilities of detecting a positive herd ranged between 52 and 99.3%, with the lowest probability for pools of 10 with 1 positive cow in the herd and the highest probability for pools of 5 with 5 positive cows in the herd. However, overall, pools of 5 and 10 had similar diagnostic capabilities, enabling cost savings by utilizing pools of 10.

Rapid assessment of bovine colostrum quality: How reliable are transmission infrared spectroscopy and digital and optical refractometers?

The objectives of this study were to evaluate the performance of the transmission infrared (IR) spectroscopic method and digital and optical Brix refractometers for measurement of colostral IgG concentration and assessment of colostrum quality of dairy cows. Colostrum samples (n = 258) were collected from Holstein cows on 30 commercial dairy farms in Nova Scotia and Newfoundland, Canada. Colostral IgG concentrations of 255 samples were measured by the reference radial immunodiffusion (RID) assay and IR spectroscopy. The Brix scores were determined on 240 of these samples using both the digital and optical Brix refractometers. Approximately half (48%) of the colostrum samples had RID IgG concentrations <50 g/L, which was the cut-point for poor quality. The correlation between RID and IR IgG concentrations was 0.88. The correlations between RID IgG concentration and Brix scores, as determined by the digital and optical refractometers, were 0.72 and 0.71, respectively. The optimal cutoff levels for distinguishing good- and poor-quality colostrum using IR spectroscopy, and digital and optical Brix refractometers were at 35 g/L and 23% Brix, respectively. The IR spectroscopy showed higher sensitivity (90%) and specificity (86%) than the digital (74 and 80%, respectively) and optical (73 and 80%, respectively) Brix refractometers for assessment of colostrum quality, as compared with RID. In conclusion, the transmission-IR spectroscopy is a rapid and accurate method for assessing colostrum quality, but is a laboratory-based method, whereas Brix refractometers were less accurate but could be used on-farm.

Identification of bovine-associated coagulase-negative staphylococci by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry using a direct transfer protocol.

This study evaluated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS) for the identification of bovine-associated coagulase-negative staphylococci (CNS), a heterogeneous group of different species. Additionally, we aimed to expand the MALDI-ToF MS database with new reference spectra as required to fill the gaps within the existing commercial spectral library. A total of 258 isolates of CNS were used in the study, covering 16 different CNS species. The majority of the isolates were previously identified by rpoB gene sequencing (n = 219), and the remainder were identified by sequencing of 16S rRNA, hsp60, or both rpoB and hsp60. The genotypic identification was considered the gold standard identification. All MALDI-ToF MS identifications were carried out using the direct transfer method. In a preliminary evaluation (n = 32 isolates; 2 of each species) with the existing commercial database, MALDI-ToF MS showed a typeability of 81% (26/32) and an accuracy of 96% (25/26). In the main evaluation (n = 226 isolates), MALDI-ToF MS with the existing commercial Biotyper (Bruker Daltonics Inc., Billerica, MA) database achieved a typeability of 92.0% (208/226) and an accuracy of 99.5% (207/208). Based on the assessment of the existing commercial database and prior knowledge of the species, a total of 13 custom reference spectra, covering 8 species, were created and added to the commercial database. Using the custom reference spectra expanded database, isolates were identified by MALDI-ToF MS with 100% typeability and 100% accuracy. Whereas the MALDI-ToF MS manufacturer's cutoff for species-level identification is 2.000, the reduction of the species level cutpoint to ≥1.700 improved the species-level identification rates (from 64 to 92% for the existing commercial database) when classifying CNS isolates. Overall, MALDI-ToF MS using the direct transfer method was shown to be a highly reliable tool for the identification of bovine-associated CNS.

Short communication: Molecular epidemiology of Streptococcus agalactiae differs between countries.

Group B Streptococcus or Streptococcus agalactiae continue to be challenging for milk quality programs in countries with emerging dairy industries, such as Colombia, where high prevalence has been reported. Molecular typing of isolates is needed to understand the variability and epidemiology of this pathogen and to develop effective control and eradication programs. We characterized the molecular profile of Strep. agalactiae isolated from cows with subclinical mastitis in 21 Colombian dairy herds and measured diversity within and between herds using multilocus sequence typing. Isolates belonged to sequence type 248 [clonal complex (CC) 103; n = 30), ST1 (CC1; n = 6) or ST22 (CC22; n = 4)], whereas members of CC67/61, the dominant type in North America, were not detected. Presence of multiple clonally unrelated sequence type within a herd was common, which contrasts with the situation in European countries and suggests introduction from multiple sources. Our results demonstrate that conclusions from molecular epidemiological studies in 1 region cannot necessarily be extrapolated to other regions, and no single bovine-adapted CC of Strep. agalactiae exists in Colombia. Improvements in internal and external biosecurity will be needed to reduce Strep. agalactiae prevalence in Colombian dairy herds.

Identifying management and disease priorities of Canadian dairy industry stakeholders.

The objective of this study was to identify the key management and disease issues affecting the Canadian dairy industry. An online questionnaire (FluidSurveys, http://fluidsurveys.com/) was conducted between March 1 and May 31, 2014. A total of 1,025 responses were received from across Canada of which 68% (n=698) of respondents were dairy producers, and the remaining respondents represented veterinarians, university researchers, government personnel, and other allied industries. Participants were asked to identify their top 3 management and disease priorities from 2 lists offered. Topics were subsequently ranked from highest to lowest using 3 different ranking methods based on points: 5-3-1 (5 points for first priority, 3 for second, and 1 for first), 3-2-1, and 1-1-1 (equal ranking). The 5-3-1 point system was selected because it minimized the number of duplicate point scores. Stakeholder groups showed general agreement with the top management issue identified as animal welfare and the number one health concern as lameness. Other areas identified as priorities were reproductive health, antibiotic use, bovine viral diarrhea, and Staphylococcus aureus mastitis with these rankings influenced by region, herd size, and stakeholder group. This is the first national comprehensive assessment of priorities undertaken in the Canadian dairy industry and will assist researchers, policymakers, program developers, and funding agencies make future decisions based on direct industry feedback.

Preliminary validation of a calf-side test for diagnosis of failure of transfer of passive immunity in dairy calves.

The objective of this study was to evaluate the utility of an initial version of a calf-side test (ZAPvet Bovine IgG test, ZBx Corp., Toronto, ON, Canada) for diagnosis of failure of transfer of passive immunity (FTPI) in dairy calves. Blood samples (n=202) were collected from calves from 1 to 11d of age. Serum IgG concentration was determined by radial immunodiffusion (RID) assay. The mean IgG concentration was 1,764±1,035mg/dL, with a range from 133 to 5,995mg/dL. The ZAPvet Bovine IgG test was used to assess FTPI (serum IgG <1,000mg/dL) and test characteristics were calculated. The number of samples that had FTPI from the RID assay and ZAPvet test was 55 and 96 samples, resulting in a true prevalence of 27% and an apparent prevalence of 47.5%, respectively. The sensitivity, specificity, and positive and negative predictive values of the ZAPvet test were 0.82, 0.65, 0.47, and 0.91, respectively. The results of the ZAPvet test were derived from 2 observers, and the overall level of agreement between the results of the 2 observers was 84%, with a kappa value of 0.67. The ZAPvet Bovine IgG test showed good potential for further development as a cost-effective, rapid calf-side test for monitoring FTPI in dairy calves.

Carryover of bovine leukemia virus antibodies in samples from shared milk meters.

Screening for infectious diseases of cattle using milk from the dairy herd improvement (DHI) sampling process is very convenient. However, when samples from shared milk meters are used, carryover of antibodies or other diagnostic targets can complicate the interpretation of the diagnostic test results for diseases, including bovine leukosis. The objectives of this study were (1) to assess the potential for carryover of antibodies against bovine leukemia virus (BLV) in milk samples obtained from shared meters, and (2) to determine if adjustment of the diagnostic test cut-off value would improve the test characteristics for meter-collected milk ELISA results. Eight dairy farms were randomly selected from herds with a wide range of BLV prevalence levels in Prince Edward Island, Canada. Within each chosen farm, 2 to 4milk meters were randomly selected. During the routine procedures of DHI sampling, 2 simultaneous milk samples, 1 hand-collected at the beginning of milking (after udder preparation) and the other from the corresponding milk meter, were taken from all lactating cows (n=236) that were milked at the selected meters (n=26). The sequence of cows using each meter was recorded. All samples were tested for BLV antibodies using a commercial indirect ELISA. Antibody carryover potential was assessed in meter-collected samples which were preceded by other cows using the same meters. Applying the hand-collected sample results as our reference standard, a new cut-off was defined for meter-collected samples to optimize the test characteristics. At the standard cut-off value of the diagnostic test, 110 (46.6%) of the hand-collected and 136 (57.6%) of the meter-collected samples were positive. For low-titer cows (e.g., true negatives), the likelihood of antibody carryover significantly increased as the titer of preceding cows increased, whereas this change was not substantial for high-titer cows. The odds of obtaining false diagnoses in meter-positive samples became larger with increase in the titer of preceding cows. A suspicious category for meter ELISA results was defined, and a retest was recommended for the cows falling into this category. This strategy effectively assisted in reducing the number of consequent false-positive results. When DHI-collected samples are used, carryover can affect the interpretation of dichotomous test results and may require adjustment of assay cut-off values.