Bovine leukemia virus causes enzootic bovine leukosis, the most common neoplastic disease of dairy cattle worldwide. Infection is lifelong, with most cattle remaining asymptomatic, approximately one-third developing persistent lymphocytosis, and a small proportion progressing to lymphoma. The proviral load (PVL), measured as the number of provirus copies per estimated white blood cell, is an indicator of transmission risk and disease progression and can be used to identify animals that contribute most to virus spread in control programs. However, longitudinal data describing PVL dynamics at the individual cow level are limited. The objectives of this study were to quantify temporal changes in PVL in naturally infected dairy cows over 3 years and evaluate the implications for repeated PVL testing in BLV control programs. A total of 716 BLV-positive cows from 25 commercial dairy herds in the Canadian Maritime provinces (Prince Edward Island, Nova Scotia, and New Brunswick) were included. Cows underwent annual PVL testing using quantitative PCR and were categorized as high, moderate, low, or undetectable PVL. Temporal changes in PVL were evaluated using repeated measurements, and conversion between PVL categories was assessed over one- and 2-year intervals. BLV proviral load showed an overall temporal trend toward higher values, with corresponding shifts in PVL categories observed across initial classification groups and within individual animals. Of cows with an initial low-PVL, 12% progressed to high-PVL over the study period. Among cows initially classified as moderate-PVL, 53% transitioned to high-PVL. A subset of cows maintained undetectable-PVL despite confirmed BLV seropositivity. These findings indicate that the likelihood of PVL progression is influenced by the initial PVL category, with moderate-PVL cows representing the group at greatest risk of becoming highly infectious. The results have practical implications for BLV control programs in dairy herds. Repeated PVL testing of cows with moderate-PVL may help identify animals at increased risk of viral transmission and guide targeted culling or segregation decisions. In contrast, cows with low-PVL may require less frequent testing, and cows with persistently undetectable-PVL may not require repeated PVL monitoring. Tailoring PVL testing frequency according to initial PVL status may improve the efficiency of BLV control programs by prioritizing management of highly infectious animals and reducing herd-level virus transmission.
IntroductionBovine Leukemia Virus (BLV) prevalence remains high in dairy cattle in North America. Quantifying the proviral load (PVL) in BLV-positive cows can be used to control this disease in herds where BLV is prevalent by focusing culling of high PVL animals to reduce the risk of transmission. The impact of high BLV PVL on dairy cows’ performance is not well established. The objective of this study was to assess the effect of high PVL status on milk production, occurrence of subclinical ketosis or mastitis, or fertility in BLV-infected cows.MethodsTwenty-five herds from the three Maritime provinces in Atlantic Canada were enrolled in this study. BLV infected cows were first identified by individual milk or serum testing. A validated quantitative qPCR was used to quantify the PVL in cows with positive BLV antibody results. Parity, 305-day milk production, annual geometric average somatic cell count, fat-to-protein ratio in milk on the first test post-calving, days in milk at first service, and calving-to-conception interval were collected from DairyComp305 software. Two-level mixed multivariable regression models were used to assess the relationship between BLV PVL and milk production, subclinical mastitis and ketosis and reproduction performance.ResultsHigh PVL was strongly associated with reduced milk production (387 kg and 431 kg) and reproduction performance (calving-to-conception interval lengthened by 50 days and 49 days), and higher odds of subclinical mastitis (Odds ratio = 2.38 and 2.48), when compared to BLVpositive cows with a low PVL and BLV-negative cows, respectively.ConclusionThese results support implementing a control program to prioritize culling high PVL cows.
Objective The primary objective was to determine the youngest age group where bovine leukemia virus (BLV)-infected dairy animals were identified. The secondary objective was to investigate associations between age-specific management practices and BLV infection status of different age groups of dairy calves and heifers. Procedure For enrolled herds, BLV status was determined using blood samples from pre-weaned calves, weaned calves, and breeding-age heifers; and bulk tank milk from the adult herd. A questionnaire investigating age-specific management factors was administered for each herd. Ordinal logistic regression was performed to identify management factors associated with the youngest age range in which BLV was identified. Results Fifty-three dairy herds from the 4 provinces in Atlantic Canada were enrolled. Bovine leukemia virus was most commonly earliest identified in pre-weaned heifers (18 herds, 32.1%) and the adult herd (18 herds, 32.1%). Ordinal logistic regression revealed that BLV was first identified in older age groups more often than in younger age groups when herds regrouped weaned heifers at least once, when fly control was used for breeding-age heifers, when herds practiced foot trimming on breeding-age heifers, and when bred heifers were brought in. Conclusion Producers can use results to identify the youngest age group(s) in which BLV is identified and to tailor management strategies to prevent new infections.
Abstract Background There is currently no commercially available method in Canada to identify bovine leukemia virus (BLV)‐positive cows with high proviral load (PVL). Objectives First, develop a model to predict PVL using common, commercially available, cost‐effective diagnostic tests. Second, investigate the relationship between lymphocyte count and PVL in BLV‐positive cows. Animals A total of 339 BLV‐positive and 62 BLV‐seronegative cows on 15 dairy farms. Methods Cross‐sectional study. Blood and milk samples were collected from all lactating BLV‐positive cows on each farm and 5 to 10 BLV‐seronegative cows depending on herd size. Blood and milk samples were tested for anti‐BLV antibodies using enzyme‐linked immunosorbent assay (ELISA). Complete blood counts were performed on blood samples, and standard components analyses were obtained for milk samples. Proviral load was determined by quantitative polymerase chain reaction for each cow. Results The inverse of lymphocyte count, the square of the inverse of lymphocyte count, and milk ELISA percent positivity were positively associated with increasing PVL in BLV‐positive cows. For BLV‐positive cows, lymphocyte count >5.2 × 109/L predicted a high PVL (BLV:Bovine DNA of >1 in blood) with a sensitivity of 92.4% and a specificity of 79.8%. For BLV‐positive cows, white blood cell count >10.8 × 109/L predicted a high PVL, with a sensitivity of 85.5% and a specificity of 83.6%. Conclusions and Clinical Importance Based on these results, producers can implement commonly available diagnostic tests to identify cows with high probability of having high PVL, which may help in designing effective disease control strategies for BLV‐positive herds.
The Atlantic Johne's Disease Initiative (AJDI) aims to control Mycobacterium avium ssp. paratuberculosis infection by using veterinary-administered risk assess-ments to identify high-risk management practices and prompt changes in management behavior. Objectives for this study were to measure producer satisfaction with the veterinary-administered risk assessment and management plan (RAMP) process in a volun-tary Johne's disease (JD) control program, compare RAMP-specific satisfaction results based on herd JD status, and measure knowledge transfer from certi-fied veterinarians to producers during the RAMP. A satisfaction questionnaire was adapted to the RAMP process in the AJDI to measure producer satisfaction. The questionnaire included 9 RAMP-specific producer satisfaction items, 1 global RAMP satisfaction item, and 16 questions to assess producer knowledge and knowledge translation about JD, bovine viral diarrhea (BVD), and bovine leukosis virus (BLV) during the RAMP (BVD and BLV used for comparison purposes). A total of 133 dairy producers in the AJDI (79.6% re-sponse rate) completed the questionnaire by telephone. The RAMP-specific satisfaction was high among the AJDI producers surveyed, and these results were not found to differ based on herd JD status. The lowest satisfaction scores and the highest number of "unable to assess" responses were for the item relating to cost. Factors that contributed to RAMP-specific producer satisfaction were not identified from the demographic and herd information available in this study. The knowledge scores indicated moderate knowledge about JD and fair knowledge about BVD and BLV. Evidence of knowledge translation from the RAMP was mixed in this study. Bovine viral diarrhea knowledge scores were not found to differ based on whether or not the certi-fied veterinarian discussed BVD during the preceding RAMP, but BLV knowledge scores were higher among dairy producers that discussed BLV during the preced-ing RAMP. Strengths and gaps in producer knowledge about these 3 infectious diseases were identified. By using this producer questionnaire, interventions aimed at improving the content, delivery, and satisfaction of RAMP in JD control programs, such as the AJDI, can be developed.
Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provides accurate species-level identification of many, microorganisms retrieved from bovine milk samples. However, not all those microorganisms are pathogenic. Our study aimed to: (1) determine the species-specific prevalence of microorganisms identified in bovine milk of apparently healthy lactating quarters vs. quarters with clinical mastitis (CM); and (2) map current information and knowledge gaps on udder health relevance of microorganisms retrieved from bovine milk samples. A mixed study design (meta-analysis and mapping review) was chosen. We gathered several large Canadian, US and Brazilian data sets of MALDI-TOF results for organisms cultured from quarter milk samples. For meta-analysis, two datasets (apparently healthy quarters vs. CM samples) were organized. A series of meta-analyses was conducted to determine microorganisms’ prevalence. Then, each species reported was searched through PubMed to investigate whether inflammation (increased somatic cell count (SCC) or signs of CM) was associated with microorganism’s recovery from milk. A total of 294 different species of microorganisms recovered from milk samples were identified. Among 50,429 quarter-milk samples from apparently healthy quarters, the 5 most frequent species were Staphylococcus chromogenes (6.7%, 95% CI 4.5–9.2%), Aerococcus viridans (1.6%, 95% CI 0.4–3.5%), Staphylococcus aureus (1.5%, 95% CI 0.5–2.8%), Staphylococcus haemolyticus (0.9%, 95% CI 0.4–1.5%), and Staphylococcus epidermidis (0.7%, 95% CI 0.2–1.6%). Among the 43,924 quarter-milk CM samples, the 5 most frequent species were Escherichia coli (11%, 95% CI 8.1–14.3%), Streptococcus uberis (8.5%, 95% CI 5.3–12.2%), Streptococcus dysgalactiae (7.8%, 95% CI 4.9–11.5%), Staphylococcus aureus (7.8%, 95% CI 4.4–11.9%), and Klebsiella pneumoniae (5.6%, 95% CI 3.4–8.2%). When conducting the PubMed literature search, there were 206 species identified by MALDI-TOF for which we were not able to find any information regarding their association with CM or SCC. Some of them, however, were frequently isolated in our multi-country dataset from the milk of quarters with CM (e.g., Citrobacter koseri, Enterococcus saccharolyticus, Streptococcus gallolyticus). Our study provides guidance to veterinarians for interpretation of milk bacteriology results obtained using MALDI-TOF and identifies knowledge gaps for future research.
In food animal production medicine (FAPM), the success of control programs for infectious diseases that have serious animal health and economic consequences frequently rely on the veterinarian's effective communication and producer adherence to veterinary recommendations. However, little research has been conducted on communication skills of practicing FAPM veterinarians. During this study, we developed a communication training workshop intervention to support the Atlantic Johne's Disease Initiative. Seventeen FAPM veterinarians across 10 clinics practicing within Maritime Canada participated in a pre-post intervention study design. Communication skills were evaluated utilizing 3 assessment tools; an objective structured clinical exam (OSCE), standardized client feedback, and an instrument designed for veterinary participants to assess their self-efficacy. Study results showed that before training, communication skills of participating veterinarians had limitations, including skill deficits in communication tasks strongly associated with increased adherence to veterinary recommendations. Based on the 3 assessment tools, communication skills of participating veterinarians improved with the training provided. Significant increases were detected in pre- to postintervention self-efficacy percentage scores, OSCE percentage and global scores from expert raters, and OSCE percentage and global scores from standardized client feedback. These improvements emphasize the importance of communication skills training specific to FAPM.
Bayesian latent class models were used to estimate the test accuracy (sensitivity (Se), specificity (Sp), and predictive values (NPV and PPV)) of cow-level somatic cell counts (SCC) data, quarter-level Petrifilm® on-farm milk culture, and quarter-level standard milk bacteriology for the identification of quarters that should possibly be treated with antimicrobials at dry off in dairy cows. Data of 282 cows from 9 dairy herds in Québec, Canada, with bulk tank SCC < 250,000 cells/mL were used. Estimated median herd-prevalence of infections that should be treated was 16.2 % (95 % credibility interval (CI): 11.0-22.7). Se and Sp estimates for quarter-milk culture using Petrifilm® were 82.2 % (95 %CI: 74.0-89.5) and 62.0 % (95 %CI: 58.6-65.6), respectively. Se and Sp for quarter-milk standard bacteriology were 67.4 % (95 %CI: 55.8-81.2) and 79.6 % (95 %CI: 76.4-83.0), respectively. Se and Sp of different SCC scenarios and thresholds were estimated. For first parity cows, using only the last Dairy Herd Improvement (DHI) test SCC with a threshold of 100,000 cells/mL appeared quite accurate, with Se, Sp, PPV, NPV and reduction of antimicrobial usage of 85.6 % (95 %CI: 69.6-95.6), 86.0 % (95 %CI: 80.0-91.7), 58.0 % (95 %CI: 42.3-74.2), 96.4 % (95 %CI: 91.3-99.0), and 75.3 % (95 %CI: 70.7-79.3), respectively. For cows of ≥ 2nd parity, using only the last DHI test SCC with a threshold of 200,000 cells/mL resulted in Se, Sp, PPV, NPV and reduction of antimicrobial usage of 75.3 % (95 %CI: 55.8-87.3), 84.0 % (95 %CI: 78.8-89.3), 47.2 % (95 %CI: 32.0-63.7), 94.7 % (95 %CI: 89.0-97.6), and of 77.0 % (95 %CI: 73.3-80.3), respectively. Adding quarter-level milk culture using Petrifilm® to cows identified as unhealthy using cow-level SCC data improved the test accuracy (mainly the PPV) and further reduced the use of antimicrobials. For instance, in ≥ 2nd parity cows, using only the last DHI SCC with a threshold of 200,000 cells/mL, adding a subsequent Petrifilm® test increased the reduction from 77.0 % (95 %CI: 73.3-80.3) to 89.5 % (95 %CI: 86.7-91.8). Considering the availability of SCC data, the easiness of using just the last DHI test, and the high NPV that could be achieved, producers may consider using just the last DHI test as a potential tool to identify cows that should be treated with antimicrobials at dry off. It may be used alone or in combination with quarter-level on-farm Petrifilm® milk culture on high SCC cows to further reduce the use of antimicrobials by identifying quarters that need to be treated.
This study was carried out to determine the antimicrobial resistance (AMR) genes and mobile genetic elements of 4 fecal blaCMY-2-producing Escherichia coli isolated from Holstein dairy calves on the same farm using whole-genome sequencing. Genomic analysis revealed that 3 of the 4 isolates shared similar genetic features, including sequence type (ST), serotype, plasmid characteristics, insertion ST, and virulence genes. In addition to genes encoding for complex multidrug resistance efflux systems, all 4 isolates were carriers of genes conferring resistance to β-lactams (blaCMY-2, blaTEM-1B), tetracyclines (tetA, tetB, tetD), aminoglycosides [aadA1, aph(3")-lb, aph(6)-ld], sulfonamides (sul2), and trimethoprim (dfrA1). We also detected 4 incompatibility plasmid groups: Inc.F, Inc.N, Inc.I, and Inc.Q. A novel ST showing a new purA and mdh allelic combination was found. The 4 isolates were likely enterotoxigenic pathotypes of E. coli, based on serotype and presence of the plasmid Inc.FII(pCoo). This study provides information for comparative genomic analysis of AMR genes and mobile genetic elements. This analysis could give some explanation to the multidrug resistance characteristics of bacteria colonizing the intestinal tract of dairy calves in the first few weeks of life.
The objective of this randomized controlled trial was to assess the efficacy of an on-farm culture system using Petrifilm (3M, London, ON, Canada) for targeted treatment decisions at the quarter level at dry-off and its effects on dry period intramammary infections (IMI) and udder health and milk production in the subsequent lactation. A total of 568 cows (2,247 quarters) from 9 dairy herds with bulk tank somatic cell count <250,000 cells/mL in Québec, Canada, were systematically enrolled and randomly allocated to 4 groups: 2 quarter-based selective (QSDCT) groups, using results of quarter-milk culture on Petrifilm, and 2 blanket dry cow therapy (BDCT) groups. The 2 QSDCT groups consisted of (1) antimicrobial to infected quarters and internal teat sealant (ITS) to healthy quarters (QSDCT/ITS); and (2) antimicrobial and ITS to infected quarters and ITS to healthy quarters (QSDCT+ITS/ITS). The 2 BDCT groups were (1) antimicrobial alone to all quarters (BDCT); and (2) antimicrobial and ITS to all quarters (BDCT+ITS). Quarter milk samples were collected at dry-off and after calving for routine bacteriological culture at the laboratory to monitor IMI; data on milk production, somatic cell count, and clinical mastitis recorded up to 120 d in milk were retrieved from health and DHI records. The probability of avoiding antimicrobial treatment in QSDCT groups was estimated at 48.3% (95% confidence interval: 35.7, 60.9). There was no significant difference between the 4 treatment groups regarding acquisition of new IMI (15.9, 13.2, 15.8, and 15.1% probability for BDCT, BDCT+ITS, QSDCT/ITS, and QSDCT+ITS/ITS, respectively) or persistence of existing IMI (3.2, 2.1, 3.4, and 2.7% probability, respectively) over the dry period. In the subsequent lactation, there was no difference between groups regarding incidence of clinical mastitis (2.4, 3.7, 2.9, and 1.7% respectively for BDCT, BDCT+ITS, QSDCT/ITS, and QSDCT+ITS/ITS), mean milk somatic cell score (1.7, 2.0, 2.0, and 2.0 respectively), or mean daily milk production (43.8, 44.2, 43.2, and 42.6 kg/d, respectively) during the first 120 d in milk. In conclusion, QSDCT using the Petrifilm on-farm culture system to detect infected quarters at dry-off is an interesting option to decrease antibiotic use without any negative effects on udder health or milk production in the first 120 d of the subsequent lactation compared with BDCT.
Over the past 30 yr, the prevalence of bovine leukemia virus (BLV) infection has increased in North America, including Atlantic Canada, at both the herd and individual cow levels. This has occurred despite increased awareness of the disease and its deleterious effects and despite implementation of management practices aimed at reducing disease transmission. Our objectives were to identify risk factors associated with the within-herd prevalence of BLV-infected cows by using a risk assessment and management program workbook, as well as to determine the current level of BLV prevalence in the Atlantic Canada region. We hypothesized that previously established risk factors, including management practices associated with calf rearing and fly control, would affect within-herd BLV prevalence. Bulk tank milk (BTM) samples were collected in January and April of 2016 and again during the same months in 2017 and 2018 from all dairy farms shipping milk in the region. Samples were tested with ELISA for levels of anti-BLV antibodies to estimate within-herd prevalence. Regional BLV prevalence at the herd level was 88.39% of dairy herds infected in 2016 and 89.30% in 2018. All dairy farms shipping milk and who had BTM samples collected in 2017 (n = 605) were eligible to participate in the risk assessment and management program questionnaire (RAMP), which was developed and distributed to all bovine veterinarians in Atlantic Canada. One hundred and six RAMP were returned, with representation from all 4 provinces. The RAMP results were combined with the mean BTM ELISA results, and univariable logistic regression followed by multivariable logistic regression was performed to investigate the association between RAMP risk factors and the estimated within-herd BLV prevalence. Factors in the multivariable model significantly associated with the odds of a herd being classified as >25% estimated within-herd prevalence included history of diagnosis of clinical BLV and calves receiving colostrum from cows with unknown BLV status. Differences in within-herd prevalence were not associated with hypodermic needle and injection practices, rectal sleeve practices, or using bulls for natural breeding, based on these 106 dairy farms.
As part of the ProAction on-farm program to implement national biosecurity requirements on dairy farms, Canadian veterinarians are called upon to complete biosecurity risk assessments. The objective of the present study was consequently to understand veterinarians' perceptions of biosecurity, and to compare perceptions among producer-veterinarian pairs. Questionnaires were administered to 106 producers from Quebec and their veterinarians (n = 82), and the answers of the producer-veterinarian pairs were compared. While most veterinarians reported it was easy to discuss biosecurity with their clients, most veterinarians and producers did not report discussing biosecurity. Moreover, when producers or veterinarians reported discussing biosecurity topics, there was no agreement amongst pairs. Veterinarians also misjudged how their clients perceived the importance and usefulness of biosecurity. Finally, veterinarians reported they had the knowledge and opportunity to assess and discuss biosecurity on dairy farms but lacked skills for evaluating farms and communicating their knowledge.
Information is scarce regarding the epidemiology of gastrointestinal nematodes in Canadian dairy heifers. The objectives of this study were to estimate the prevalence and fecal egg counts of gastrointestinal nematodes in dairy heifers, and using a novel deep-amplicon sequencing approach, to identify the predominant gastrointestinal nematode species in Canadian dairy replacement heifers. Fresh environmental fecal samples (n = 2,369) were collected from replacement heifers on 306 dairy farms across western Canada, Ontario, Québec, and Atlantic Canada. Eggs per gram of feces (EPG) were determined using a modified Wisconsin double-centrifugation sugar flotation technique. Predominant nematode species at the farm level were identified by deep-amplicon nemabiome sequencing of the internal transcribed spacer-2 rDNA locus of nematode third-stage larvae. Generalized estimating equations were used to estimate predicted parasite prevalence and mean EPG in all heifers and by province, allowing for clustering within herds. Individual heifer egg counts ranged from 0 to 141 EPG (median: 0 EPG; interquartile range: 0 to 71 EPG). Gastrointestinal nematodes were detected in 20.9% (95% confidence interval: 17.2 to 24.6%) of heifers, and the predicted mean strongylid EPG accounting for clustering on farms was 1.1 (95% confidence interval: 0.6 to 1.6). The predominant parasite species were Cooperia oncophora and Ostertagia ostertagi. This is the first study in Canada to use a combination of deep-amplicon nemabiome sequencing and a traditional egg count method to describe the epidemiology of gastrointestinal nematodes in dairy heifers.
The primary objective of this observational study was to examine the association between passive transfer of immunity and growth performance in preweaning calves. A secondary objective was to evaluate the utility of a heart girth tape (HGT) to estimate body weight (BW) in preweaning calves. A total of 142 Holstein calves were enrolled in this study. Blood samples were collected 24 to 48 hours after birth and serum immunoglobulin G (IgG) concentration for each calf was measured by radial immunodiffusion assay. Calf BW was determined at birth, at 21 days, and at weaning using an electronic scale (ES) and HGT. A significant positive association was detected between serum IgG and both BW at 21 days and average daily gain (ADG) from 0 to 21 days of life. Additionally, ADG from 0 to 42 days of life showed a trend toward an improved rate of gain as IgG concentration increased. The Pearson correlation coefficient between BW obtained from ES and HGT was 0.81 at birth, 0.86 at 21 days, and 0.83 at weaning. The mean differences between BW obtained from ES and HGT were -3.1 kg at birth, -3.2 kg at 21 days, and -7.7 kg at weaning. In conclusion, serum IgG concentration in neonatal calves is an important contributing factor for the variation in growth performance of preweaning calves. The HGT can be used to estimate the BW of preweaning calves but has a tendency to overestimate weight, especially at weaning compared to birth and 21 days of age.
Accurate diagnosis of failure of transfer of passive immunity (FTPI) in newborn calves is an essential component of dairy farm management plan. Several methods (direct and indirect) are available for diagnosis of FTPI in dairy calves. However, the indirect methods offer an advantage over the direct methods in not requiring an experienced veterinarian, rapid, cost efficient and can be performed under field-setting. The objective of this study was to estimate the diagnostic performance of radial immunodiffusion (RID) assay, transmission infrared (TIR) spectroscopy and digital Brix refractometer for diagnosis of FTPI in dairy calves using latent class models at four cut-off values of digital Brix refractometer. Holstein calves (n = 691) from 40 commercial dairy farms in the four Atlantic Canada provinces were blood-sampled and tested for detection of FTPI. Results showed that the number of calves with FTPI was 253 (36.6%) by RID, 194 (28.1%) by TIR and 204 (293%) by Brix refractometer at cut-off value of 8.2%. Estimates of Se-RID was higher than Se-TIR and Se-Brix, at all Brix refractometer cut-offs, but with increase of Brix refractometer cut-off from 8.2 to 8.5%, Se-RID and Se-TIR were decreased from 96.0% (95% PCI: 88.0-99.0) and 79.0% (95% PCI: 70.0-85.0), to 92.0% (95% PCI: 77.0-99.0) and 74.0% (95% PCI: 61.0-82.0), respectively. SPRID and Sp(TIR) were always higher than Sp(Brix) at all tested cut-offs and were above 92.0%, and 96.0%, respectively. With increasing the cut-off of Brix refractometer from 8.2 to 8.5%, Se-Brix estimate has remarkably increased from 79.0% (95% PCI: 70.0-96.0) to 95.0% (95% PCI: 87.0-100.0), respectively. Whilst, Sp(Brix) was decreased from 95.0% (95% PCI: 91.0-98.0) at cut-off 8.2% to 84.0% (95% PCI: 78.0-94.0) at cut-off 8.5%. In conclusion, RID has a higher Se than TIR and Brix, if the latter is used with cut-offs of 8.2% or 8.3%. However, the higher the cut-off, the more comparable sensitivities of RID and digital Brix refractometer. The median estimate of Sp(TIR) was always higher than Sp(RID) and Sp(Brix) at all tested cut-offs. However, the 95% confidence interval estimates of the three tests were overlapping across the tested cut-offs of digital Brix refractometer reflecting the inability to prefer a test over the other based on the Sp estimate.
Staphylococcus aureus is a bacterial pathogen causing bovine intramammary infections (IMIs) often leading to chronic clinical or subclinical mastitis. Predicting the outcome of S. aureus IMIs (duration and clinical vs subclinical) based on the characterization of isolates would help to make better case management decisions. For this purpose, 583 S. aureus isolates from series of quarter milk samples were characterized by genotypic tests (detection of virulence genes seg, tst, lukM), epidemiological typing (spa type) and by a phenotypic test (biofilm production). VNTR typing (variable number of tandem repeats) was used to establish persistence of the same S. aureus strain in each series of sequential isolates. This allowed to associate each strain to a clinical/subclinical status and to validate the duration of infection. We found differences in the distribution of spa types between the strains from clinical and subclinical cases. Prevalence of lukM was also higher in strains from clinical cases than in strains from subclinical cases. A Kaplan-Meier analysis was then used to determine factors influencing the duration of the infection. Considering a multivariable model of the logistic regression, time to elimination was shorter with the strains of the subclinical lactation series compared to the clinical series (series with at least one clinical case). Strains from the spa type t359 and t529 were less likely to persist compared to those of spa type t13410. In sum, strain characterization including determination of the spa type helps to predict duration of infection and the clinical or subclinical outcome of S. aureus IMIs.
The objective of this study was to explore the potential of transmission infrared (TIR) spectroscopy in combination with partial least squares regression (PLSR) for quantification of dairy and beef cow colostral immunoglobulin G (IgG) concentration and assessment of colostrum quality. A total of 430 colostrum samples were collected from dairy (n = 235) and beef (n = 195) cows and tested by a radial immunodiffusion (RID) assay and TIR spectroscopy. Colostral IgG concentrations obtained by the RID assay were linked to the preprocessed spectra and divided into combined and prediction data sets. Three PLSR calibration models were built: one for the dairy cow colostrum only, the second for beef cow colostrum only, and the third for the merged dairy and beef cow colostrum. The predictive performance of each model was evaluated separately using the independent prediction data set. The Pearson correlation coefficients between IgG concentrations as determined by the TIR-based assay and the RID assay were 0.84 for dairy cow colostrum, 0.88 for beef cow colostrum, and 0.92 for the merged set of dairy and beef cow colostrum. The average of the differences between colostral IgG concentrations obtained by the RID- and TIR-based assays were -3.5, 2.7, and 1.4 g/L for dairy, beef, and merged colostrum samples, respectively. Further, the average relative error of the colostral IgG predicted by the TIR spectroscopy from the RID assay was 5% for dairy cow, 1.2% for beef cow, and 0.8% for the merged data set. The average intra-assay CV% of the IgG concentration predicted by the TIR-based method were 3.2%, 2.5%, and 6.9% for dairy cow, beef cow, and merged data set, respectively.The utility of TIR method for assessment of colostrum quality was evaluated using the entire data set and showed that TIR spectroscopy accurately identified the quality status of 91% of dairy cow colostrum, 95% of beef cow colostrum, and 89% and 93% of the merged dairy and beef cow colostrum samples, respectively. The results showed that TIR spectroscopy demonstrates potential as a simple, rapid, and cost-efficient method for use as an estimate of IgG concentration in dairy and beef cow colostrum samples and assessment of colostrum quality. The results also showed that merging the dairy and beef cow colostrum sample data sets improved the predictive ability of the TIR spectroscopy.
This study was carried out to determine the frequency of fecal carriage, antimicrobial susceptibility, and resistance genes in Salmonella enterica and Escherichia coli with reduced susceptibility to extended-spectrum cephalosporins (ESC) isolated from 488 dairy calves from 8 farms in New Brunswick, Canada. Both S. enterica and E. coli with reduced susceptibility to ESC were isolated using selective culture. Minimum inhibitory concentrations to a panel of antimicrobial drugs were determined for randomly selected E. coli isolates and all of the Salmonella isolates. Multiplex PCR were conducted on the selected ESC-resistant E. coli to assess the β-lactamase resistance genes (blaCTX-M, blaCMY-2, blaSHV, and blaTEM) and plasmid-mediated qnrB and qnrS resistant genes. Information on ceftiofur use and other farm management practices were collected by the use of a questionnaire to determine the risk factors for the fecal recovery of E. coli with reduced susceptibility to ESC. Salmonella enterica frequency in calves' fecal samples was 3.3%, and all were pansusceptible. Salmonella isolates belonged to 3 serovars namely Salmonella Senftenberg, Salmonella Typhimurium, and Salmonella Derby. The frequency of fecal carriage of E. coli with reduced susceptibility to ESC in calves was 81.2%. Of the selected isolates (n = 100), all were multi-drug resistant, whereas 88% were ESC resistant based on minimum inhibitory concentration testing. From the selected ESC-resistant E. coli isolates, blaTEM was detected in 84.1%, blaCMY-2 was detected in 52.2%, blaCTXM groups were detected in 30.7%, and blaSHV was detected in 1.1% of isolates. Plasmid-mediated quinolone resistance genes were identified in 7 of 9 isolates resistant to quinolones. Five isolates were positive for qnrB, whereas 2 isolates were positive for both qnrB and qnrS. Whereas neonatal calves [odds ratio (OR) = 2.42, 95% confidence interval (CI): 1.87-3.12], regular ceftiofur use on the farm (OR = 3.83, 95% CI: 2.29-6.39), feeding of unpasteurized nonsalable milk (OR = 1.6, 95% CI: 1.18-2.18), and use of florfenicol (OR = 2.02, 95% CI: 1.43-2.86) were statistically associated with fecal recovery of E. coli with reduced susceptibility to ESC, use of ceftiofur for the treatment of respiratory diseases (OR = 0.57, 95% CI: 0.41-0.79) was statistically associated with decreased recovery of E. coli with reduced susceptibility to ESC. This study has provided information on the resistance genes associated with the occurrence of ESC and fluoroquinolone resistance in dairy calves within this region.
In the past decade, substantial changes have occurred in the way dairy heifer calves are managed. The objectives of this study, part of phase I of the 2015 Canadian National Dairy Study, were to examine heifer calf health and adoption of rearing practices, and to explore factors associated with different rearing strategies on Canadian dairy farms. The questionnaire was open to all licensed dairy producers in Canada and had a 12% response rate (n = 1,373). Fifty-one percent of respondents reported never allowing heifer calves to nurse their dam, and 17% always removed calves within 30 min of birth. Sixty-seven percent reported always feeding heifer calves 4 L of colostrum within 12 h of birth; 17% always fed colostrum between 2100 and 0400 h; 5% pasteurized all colostrum fed on farm. Seventy-seven percent of respondents reported calving and stillbirth information for 2014; the mean reported stillbirth rate was 4.9% (SD = 3.3). Forty percent housed calves in individual pens, 34% in group pens, 21% in individual hutches, 2% reported tethering calves, and 1% used group hutches. Of those who housed calves in groups, 59% reported a maximum group size of 3 to 10 calves, 31% reported a pair (2) of calves per pen or hutch, and 10% reported a group >10 calves. The maximum amount of milk offered to calves per day during the preweaned period was a mean of 8 L (SD = 3). Fifty percent of respondents reported mortality data for 2014; mean preweaning mortality was 6.4% (SD = 8.3), and postweaning mortality was 2.4% (SD = 4.4). Over 95% of producers reported disbudding practices; 86% used cautery, 11% used surgical amputation, and 9% used caustic paste. Twenty-eight percent reported disbudding at less than 3 wk of age, 60% at 3 to 8 wk, and 22% at 8 to 16 wk; 5% of respondents reported dehorning at more than 16 wk of age. Sixty-six percent of cautery users reported use of local anesthetic, 33% used sedation, and 25% used a nonsteroidal anti-inflammatory drug (NSAID). Multivariable regression models showed that the use of local anesthetic when disbudding calves was associated with social media activity (odds ratio = 2.3) and high-speed internet access (odds ratio = 2.0), whereas sedation was associated with geographic region, and NSAID use was associated with disbudding at more than 3 wk of age. Exploring heifer rearing management practices, including adoption of best practices, may help focus future education and extension efforts. Poor reporting of mortality data may reflect a lack of recordkeeping on farm.