J.-P. Roy

Manageable risk factors associated with the lactational incidence, elimination, and prevalence of Staphylococcus aureus intramammary infections in dairy cows

Staphylococcus aureus intramammary infections (IMI) are a major cause of mastitis on farms worldwide. Incidence and elimination rates are the key determinants of prevalence of Staph. aureus, and risk factors associated with these rates must be identified, prioritized, and controlled to obtain long-term reduction in prevalence. The objectives of this study were to identify manageable risk factors associated with the lactational incidence, elimination, and prevalence of Staph. aureus IMI. A cohort of 90 Canadian dairy farms was recruited and followed in 2007 and 2008. Quarter milk samples were collected repeatedly from a selection of cows, and bacteriological culture was realized to assess incidence, elimination, and prevalence of Staph. aureus IMI. Practices used on farms were measured using direct observations and a validated questionnaire. A linear regression model was used to explore the relationship between herd IMI prevalence and incidence and elimination rates. Multilevel logistic regression models were used to compute measures of associations between practices used on farms and IMI incidence, elimination, and prevalence. The herd incidence rate was the most important predictor of herd IMI prevalence: a reduction of the incidence rate equivalent to its interquartile range (0.011 new IMI/quarter-month) was associated with a prevalence reduction of 2.2 percentage points; in comparison, an equivalent increase of the elimination rate by its interquartile range (0.36 eliminated IMI/quarter-month) resulted in a prevalence reduction of 0.4 percentage points. Postmilking teat disinfection and blanket dry-cow therapy were already implemented by most herds. Most of the practices associated with Staph. aureus IMI incidence were related to milking procedures. Among these, wearing gloves during milking showed desirable associations with IMI incidence, elimination, and prevalence. Similarly, adequate teat-end condition and use of premilking teat disinfection were associated with lower IMI incidence and prevalence. The initial herd prevalence of Staph. aureus IMI was positively associated with subsequent IMI incidence. This indicates that, in some situations, an initial reduction of the pool of infected quarters could be justified. Some housing practices were associated with IMI incidence, elimination, or prevalence. The effects of these latter practices, however, were often influenced by specific cow characteristics such as parity or days in milk. These results highlight the importance of good milking practices to prevent Staph. aureus IMI acquisition and, therefore, reduce their prevalence.

Epidemiology of coagulase-negative staphylococci intramammary infection in dairy cattle and the effect of bacteriological culture misclassification.

Objectives of this study were to identify the manageable risk factors associated with the lactational incidence, elimination, and prevalence of coagulase-negative staphylococci (CNS) intramammary infections (IMI) while taking into account the difficulties inherent to their diagnosis. A second objective was to evaluate the effect of CNS IMI misclassification in mastitis research. A cohort of 90 Canadian dairy herds was followed throughout 2007 to 2008. In each herd, series of quarter milk samples were collected from a subsample of cows and bacteriological culture was performed to identify prevalent, incident, and eliminated CNS IMI. Practices used on farms were captured using direct observations and a validated questionnaire. The relationships between herd CNS IMI prevalence and herd incidence and elimination rates were explored using linear regression. Manageable risk factors associated with the prevalence, incidence, or elimination of CNS IMI were identified via Bayesian analyses using a latent class model approach, allowing adjustment of the estimates for the imperfect sensitivity and specificity of bacteriological culture. After adjustment for the diagnostic test limitations, a mean CNS IMI quarter prevalence of 42.7% [95% confidence interval (CI): 34.7, 50.1] and incidence and elimination rates of 0.29 new IMI/quarter-month (95% CI: 0.21, 0.37) and 0.79 eliminated IMI/quarter-month (95% CI: 0.66, 0.91), respectively, were observed. Considerable biases of the estimates were observed when CNS IMI misclassification was ignored. These biases were important for measures of association with risk factors, were almost always toward the null value, and led to both type I and type II errors. Coagulase-negative staphylococci IMI incidence appeared to be a stronger determinant of herd IMI prevalence than IMI elimination rate. The majority of herds followed were already using blanket dry cow treatment and postmilking teat disinfection. A holistic approach considering associations with all 3 outcomes was used to interpret associations between manageable risk factors and CNS IMI. Sand and wood-based product bedding showed desirable associations with CNS IMI compared with straw bedding. Quarters of cows that had access to pasture during the sampling period had lower odds of acquiring a new CNS IMI and of having a prevalent CNS IMI. Many practices showed an association with only one of the CNS outcomes and should, therefore, be considered with caution.

Effect of nonclinical Staphylococcus aureus or coagulase-negative staphylococci intramammary infection during the first month of lactation on somatic cell count and milk yield in heifers

Coagulase-negative staphylococci (CNS) are the most prevalent cause of intramammary infections in heifers around calving, but Staphylococcus aureus should not be ignored because it is also prevalent, contagious, and more likely to persist into lactation. The objective of this study was to determine the effect of a subclinical infection caused by S. aureus or CNS diagnosed during the first month of lactation in heifers on SCC, milk production, and culling risk during the entire first lactation. Data were obtained from a cohort of 50 farms following a mastitis monitoring and control program and subscribing to the animal health record system (DS@HR) through the ambulatory clinic of the Faculté de médecine vétérinaire of the Université de Montréal (St-Hyacinthe, Québec, Canada). This program included routinely collecting a composite milk sample at each farm visit from all recently freshened heifers. A total of 2,273 Holstein heifers were examined. Among the 1,691 heifers meeting the full selection criteria, 90 (5%) were diagnosed with S. aureus, 168 (10%) were diagnosed with CNS, and 153 (9%) were negative (no pathogen isolated). Test-day natural logarithm somatic cell count (lnSCC) was modeled in a repeated measures linear regression model with herd as random effect. The model-adjusted mean lnSCC in S. aureus and CNS groups were significantly higher than in the culture-negative group from 40 to 300 d in milk. At the test-day level, lnSCC in S. aureus and CNS groups were on average 1.2 and 0.6 higher, respectively, than the culture-negative group. A similar model for milk yield showed that mean milk yield was not statistically different between culture groups from 40 to 300 d in milk. The presence of a S. aureus or CNS intramammary infections in the first month of lactation in heifers correlates with future increased SCC over the entire first lactation.

Genotypic and phenotypic characterization of Staphylococcus aureus causing persistent and nonpersistent subclinical bovine intramammary infections during lactation or the dry period

Staphylococcus aureus is a significant pathogen frequently causing persistent intramammary infections (IMI) in dairy cows. We compared some genotypic and phenotypic characteristics of 285 strains collected from quarter milk samples from cows with persistent and nonpersistent subclinical IMI across Canada. Variable number of tandem repeats typing was used to infer the persistence of the same S. aureus strain in 3 consecutive quarter milk samples collected at intervals of 3 wk during lactation or before and after dry-off. All first isolates of the series were used as the representative strains from persistent IMI and were compared with nonpersistent strains for the presence of genes seg, sen, sec, and tst as well as by spa typing. Biofilm production in vitro and hld-RNAIII expression levels were also quantified. The gene seg was associated with a reduction in the likelihood of the bacteria to cause a persistent IMI during lactation. Strains persisting through the dry period produced significantly more biofilm in vitro than strains that do not persist after calving. Also, we showed that strains expressing more hld were more likely to be nonpersistent during either lactation or through the dry period. Three spa types were predominant (t529, t267, and a novel type: t13401). In the strains studied, the spa type tbl 2645 was the most frequent, and 97.0% of the strains of this spa type carried both sen and seg. Strains from the spa type tbl 2645 were less likely to cause a persistent IMI in the dry period. Most (86.7%) of the strains of the novel spa type (t13401) were negative for seg, sen, or both and produced significantly more biofilm in vitro than tbl 2645 and t267. The present study expanded our current knowledge on the genotypic and phenotypic traits of S. aureus strains recovered from persistent and nonpersistent IMI in Canada.

Evaluation of selective dry cow treatment following on-farm culture: Milk yield and somatic cell count in the subsequent lactation

Compared with blanket dry cow therapy (DCT), the selective antimicrobial treatment of cows based upon on-farm culture results has the potential to reduce the amount of antimicrobials used in dairy production. The objective of the current study was to determine the effect of a Petrifilm (3M Canada, London, Ontario) on-farm culture-based selective DCT program on milk yield and somatic cell count (SCC) in the following lactation. A total of 729 low-SCC (<200,000 cells/mL) cows from 16 commercial dairy herds with a low bulk tank SCC (<250,000 cells/mL) were randomly assigned to receive either blanket DCT or Petrifilm-based selective DCT. Cows belonging to the blanket DCT group were infused with a commercial DCT product and an internal teat sealant (ITS) at drying off. Using composite milk samples collected on the day before drying off, cows in the selective DCT group were treated at drying off based on the results obtained by the Petrifilm on-farm culture system with DCT and ITS (Petrifilm culture positive) or ITS alone (Petrifilm culture negative). Milk test-day records for the following lactation were obtained from Dairy Herd Improvement for all cows enrolled in the trial. Repeated measures linear mixed models were used to assess the effect of study group (blanket or selective DCT) on test-day milk production and natural logarithm of SCC over the first 180 d of the subsequent lactation. According to the final multivariable models, when low-SCC cows were selectively treated with DCT at drying off based on results obtained using the Petrifilm on-farm culture system, no effect on milk production (least squares means for blanket DCT = 39.3 kg vs. selective DCT = 39.0 kg) or natural logarithm of SCC (least squares means for blanket DCT = 3.95 vs. selective DCT = 3.97) was observed in the subsequent lactation when compared with cows receiving blanket DCT. The results of this study indicate that selective DCT based on results obtained by the Petrifilm on-farm culture system enabled a reduction in the use of DCT without negatively affecting milk production and milk quality.

Increased blood-circulating interferon-γ, interleukin-17, and osteopontin levels in bovine paratuberculosis.

Paratuberculosis-infected cattle initially develop an effective cell-mediated immune response that declines as the disease progresses. Blood is one of best sources for characterizing the inflammatory status of infected cows and for studying mediators related to chronic diseases. The aim of this study was to evaluate the cow-level association between blood cytokine concentration, the influence of serum on immune cell proliferation, and dairy cows naturally infected with Mycobacterium avium ssp. paratuberculosis (MAP). Positive animals (n=41) from 19 herds were selected on the basis of 2 positive fecal culture results and divided into 2 groups: single-positive, or serum ELISA-negative cows (n=32), and double-positive, or cows that gave positive results for both mycobacterial culture and serum ELISA (n=9). Negative animals (n=39) were selected from paratuberculosis-negative herds in which at least 80% of the animals had been diagnosed as negative by fecal culture and ELISA and that did not produce positive results during the 2-yr study. Analysis of plasma levels of the cytokines IL-4, IL-10, IL-17, IFN-γ, and osteopontin was performed, revealing distinct patterns. The ELISA-positive cows with MAP shedding had similar plasma concentrations of IL-4 and IL-10 but elevated levels of IFN-γ, IL-17, and osteopontin, which is indicative of inflammatory disease in these subclinical positive cows. In vitro MAP infection of bovine macrophages showed increased gene expression of tumor necrosis factor-α, IL-1β, IL-6, IL-23, and transforming growth factor-β as early as 6h postinfection for all of the cytokines involved in the establishment of a T-helper type-17 immune response. To determine the systemic influence of serum on immune cell functions, lymphoproliferation assays were also performed in presence of JD serum. The serum from shedding cows showed 15% less proliferation. These results indicate that infected cows have a lower systemic capacity to maintain a protective immune response and that, as the disease progresses, an emerging T-helper type-17 immune response is established.

A randomized controlled trial on the effect of incomplete milking during early lactation on ketonemia and body condition loss in Holstein dairy cows

Limiting milk production for a short period of time in early lactation could be a relevant strategy to prevent hyperketonemia (HYK). From December 2013 to March 2015, 838 multiparous Holstein cows from 13 herds were enrolled in a randomized controlled trial evaluating the effect of incomplete milking in early lactation on ketonemia and its effect on body condition score (BCS) loss. Cows were randomly assigned 4 wk before expected calving date to 1 of 2 treatment groups, (1) a conventional milking protocol (CON) for which cows were completely milked or (2) an incomplete milking protocol (INC) for which a maximum of 10 to 14 kg of milk/d were withdrawn during the first 5 d in milk (DIM). β-Hydroxybutyrate (BHB) concentrations were measured from blood samples collected on each cow 3 times at weekly intervals. Hyperketonemia was defined as BHB ≥1.4 mmol/L. Body condition score variation in the postcalving period was calculated by subtracting BCS assessed at wk 7 from BCS assessed at first week after calving. Effect of treatment on ketonemia and prevalence of HYK were evaluated for 4 specific time periods: 1 to 3, 4 to 7, 8 to 17, and 18 to 26 DIM. Effect of treatment on ketonemia was investigated using linear mixed models with natural logarithm of BHB measurements as outcome and treatment groups as fixed effect. Generalized linear mixed models with HYK as outcome, using logit link, and treatment groups as fixed effect were used to investigate effect of treatment on odds of HYK. A logistic regression model with BCS loss (<0.75 or ≥0.75) as outcome and treatment groups and herd as fixed effects was used to study effect of INC on odds of having BCS loss ≥0.75. A total of 813 lactations had complete data and were used for statistical analysis of ketonemia and HYK. A total of 709 lactations had complete data and were used for analysis of BCS loss. Geometric means of blood BHB concentrations during the 1 to 3, 4 to 7, 8 to 17, and 18 to 26 DIM periods were, respectively, 0.72 (95% confidence interval = 0.66, 0.80), 0.66 (0.60, 0.73), 0.90 (0.80, 1.01), and 0.93 (0.83, 1.05) mmol/L for INC, and 0.65 (0.59, 0.72), 0.79 (0.72, 0.87), 0.94 (0.84, 1.06), and 0.92 (0.82, 1.04) mmol/L for CON. Cows in INC group had lower ketonemia during the 4 to 7 DIM period. Predicted prevalence of HYK during the 1 to 3, 4 to 7, 8 to 17, and 18 to 26 DIM periods were, respectively, 2.8 (3.2, 15.1), 4.6 (2.0, 10.0), 13.4 (8.4, 20.0), and 23.0% (17.4, 29.7) for INC and 2.6 (2.5, 13.8), 10.7 (5.6, 19.3), 19.4 (13.0, 27.9), and 21.3% (16.0, 27.8) for CON. The INC treatment reduced the prevalence of HYK during the 4 to 7 and 8 to 17 DIM periods. No association was observed between INC and BCS loss in the postcalving period. Overall, the incomplete milking protocol was effective for reducing ketonemia and prevalence of HYK during the early postpartum period.

Invited review: A systematic review and qualitative analysis of treatments other than conventional antimicrobials for clinical mastitis in dairy cows

Clinical mastitis is an important disease in dairies. Its treatment is mainly based on the use of antimicrobial drugs. Numerous non-antimicrobial drugs and treatment strategies have already been reported for clinical mastitis treatment, but data on their efficacy have never been collated in a systematic way. The objective of this systematic review was to identify treatments other than conventional antimicrobials for the treatment of clinical mastitis in lactating dairy cows. A systematic review was performed with studies written in English or French selected from CAB Abstracts, PubMed, and Web of Science from January 1970 to June 2014. Controlled clinical trials, observational studies, and experimental challenges were retained. Lactating dairy cows with clinical mastitis were the participant of interest. All treatments other than conventional antimicrobials for clinical mastitis during lactation were retained. Only studies comparing the treatment under investigation to a negative or positive control, or both, were included. Outcomes evaluated were clinical and bacteriological cure rates and milk production. Selection of the study, data extraction, and assessment of risk of bias was performed by 3 reviewers. Assessment of risk of bias was evaluated using the Cochrane Collaboration tool for systematic review of interventions. A total of 2,451 manuscripts were first identified and 39 manuscripts corresponding to 41 studies were included. Among these, 22 were clinical trials, 18 were experimental studies, and 1 was an observational study. The treatments evaluated were conventional anti-inflammatory drugs (n = 14), oxytocin with or without frequent milk out (n = 5), biologics (n = 9), homeopathy (n = 5), botanicals (n = 4), probiotics (n = 2), and other alternative products (n = 2). All trials had at least one unclear or high risk of bias. Most trials (n = 13) did not observe significant differences in clinical or bacteriological cure rates in comparison with negative or positive controls. Few studies evaluated the effect of treatment on milk yield. In general, the power of the different studies was very low, thus precluding conclusions on noninferiority or nonsuperiority of the treatments investigated. No evidence-based recommendations could be given for the use of an alternative or non-antimicrobial conventional treatment for clinical mastitis. However, probiotics and oxytocin with or without frequent milk out should not be recommended. We concluded that homeopathic treatments are not efficient for management of clinical mastitis.

A randomized controlled trial on the effect of incomplete milking during the first 5 days in milk on culling hazard and on milk production and composition of dairy cows

An incomplete milking in early lactation could help limit negative energy balance in dairy cattle, but its potential effects on culling hazard and on milk production and composition throughout the entire lactation are unknown. The objective of this study was to evaluate the effect of an incomplete milking during the first 5 d in milk on culling hazard, milk weight, milk fat and protein concentrations, and energy-corrected milk (ECM) yield during the whole lactation. A randomized controlled trial was conducted in 13 dairy farms near St-Hyacinthe, Quebec, Canada. Approximately 1 mo before expected calving, Holstein multiparous cows calving between December 2013 and March 2015 (n = 846 cow lactations) were randomly assigned to a control or a treatment group. Cows in the control group were milked conventionally, whereas cows in the treatment group were submitted to an incomplete milking protocol (maximum of 10, 12, and 14 L/d of milk was collected on days in milk 1-3, 4, and 5, respectively). All farms were registered on Dairy Herd Improvement Association, which was used to obtain records on culling, monthly milk yield, and milk fat and protein concentrations. In addition, daily milk yield records were available for 6 farms. A Cox proportional hazards model with a herd frailty term was fitted to the data to compare culling hazard among treatment groups. Regarding milk production and composition, 4 linear mixed models with herd as a fixed effect, cow as a random effect, and using an autoregressive covariance structure were used to study the effect of the incomplete milking on (1) milk weight, (2) milk fat concentration, (3) milk protein concentration, and (4) ECM yield. Culling hazard did not differ among treatment groups (hazard ratio = 1.0; 95% CI = 0.82, 1.3). We observed no differences in milk weight, milk fat, or protein concentration among treatment groups between weeks in milk (WIM) 2 and 44 (the studied period). We noted a difference in ECM between treatment groups for WIM 38, with incompletely milked cows producing less milk than conventionally milked cows (-2.7 kg/d; 95% CI = -0.02, -5.2 kg/d), but no differences were found for any of the other WIM. These results suggest that this strategy for controlling the negative energy balance has negligible effect on cow productivity.

Short communication: An observational study investigating inter-observer agreement for variation over time of body condition score in dairy cows

Body condition score (BCS) is strongly correlated with energy reserves. The ease, rapidity of scoring, and high intra- and inter-observer repeatability make it a widely used herd management tool in bovine practice and in scientific studies. Loss or gain of BCS, rather than a single BCS measurement, is frequently used to monitor energy balance in dairy cows. It is unknown if the difference between 2 BCS measures taken at different moments (ΔBCS) would demonstrate inter-observer agreement similar to that of a single BCS measurement. The objective of this study was to compare inter-observer agreement of BCS and ΔBCS in dairy cows when multiple observers perform data collection. An observational study was conducted between April and September 2015; 3 observers independently assessed BCS of 73 Holstein cows from 1 commercial dairy herd. Body condition score assessments of the animals were performed between 1 and 20 d in milk (early lactation; exam 1) and again between 41 and 60 d in milk (peak of milk production; exam 2). Quadratic weighted kappa (κw) was computed to quantify agreement between observers for single BCS measurements and ΔBCS. For single BCS measurements, κw of 0.79 (95% CI: 0.69, 0.85) and 0.84 (95% CI: 0.77, 0.89) were obtained for exam 1 and exam 2, respectively. Such values would be interpreted as strong agreement and are consistent with the available literature on BCS repeatability. When computing agreement for ΔBCS, a κw value of 0.49 (95% CI: 0.32, 0.63) was obtained, suggesting moderate agreement between observers. These findings suggest that studies investigating single BCS measures could use many observers with a high degree of accuracy in the results. When ΔBCS is the parameter of interest, more reliable results would be obtained if one observer conducts all assessments.