No nationwide studies of the incidence rate of clinical mastitis (IRCM) have been conducted in Canada. Because the IRCM and distribution of mastitis-causing bacteria may show substantial geographic variation, the primary objective of this study was to determine regional pathogen-specific IRCM on Canadian dairy farms. Additionally, the association of pathogen-specific IRCM with bulk milk somatic cell count (BMSCC) and barn type were determined. In total, 106 dairy farms in 10 provinces of Canada participated in the study for a period of 1 yr. Participating producers recorded 3,149 cases of clinical mastitis. The most frequently isolated mastitis pathogens were Staphylococcus aureus, Escherichia coli, Streptococcus uberis, and coagulase-negative staphylococci. Overall mean and median IRCM were 23.0 and 16.7 cases per 100 cow-years in the selected herds, respectively, with a range from 0.7 to 97.4 per herd. No association between BMSCC and overall IRCM was found, but E. coli and culture-negative IRCM were highest and Staph. aureus IRCM was lowest in low and medium BMSCC herds. Staphylococcus aureus, Strep. uberis, and Streptococcus dysgalactiae IRCM were lowest in the Western provinces. Staphylococcus aureus and Strep. dysgalactiae IRCM were highest in Québec. Cows in tie-stalls had higher incidences of Staph. aureus, Strep. uberis, coagulase-negative staphylococci, and other streptococcal IRCM compared with those in free-stalls, whereas cows in free stalls had higher Klebsiella spp. and E. coli IRCM than those in tie-stall barns. The focus of mastitis prevention and control programs should differ between regions and should be tailored to farms based on housing type and BMSCC.

Antimicrobial use (AMU) data are critical for formulating policies for containing antimicrobial resistance. The present study determined AMU on Canadian dairy farms and characterized variation in AMU based on herd-level factors such as milk production, somatic cell count, herd size, geographic region and housing type. Drug use data were collected on 89 dairy herds in 4 regions of Canada, Alberta, Ontario, Québec, and the Maritime provinces (Prince Edward Island, New Brunswick, and Nova Scotia) for an average of 540 d per herd. Dairy producers and farm personnel were asked to deposit empty drug containers into specially provided receptacles. Antimicrobial use was measured as antimicrobial drug use rate (ADUR), with the unit being number of animal defined-daily doses (ADD)/1,000 cow-days. Antimicrobial drug use rates were determined at farm, region, and national level. Combined ADUR of all antimicrobial classes was 14.35 ADD/1,000 cow-days nationally. National level ADUR of the 6 most commonly used antimicrobial drug classes, cephalosporins, penicillins, penicillin combinations, tetracyclines, trimethoprim-sulfonamide combinations, and lincosamides were 3.05, 2.56, 2.20, 1.83, 0.87, and 0.84 ADD/1,000 cow-days, respectively. Dairy herds in Ontario were higher users of third-generation cephalosporins (ceftiofur) than in Québec. Alberta dairy herds were higher users of tetracyclines in comparison to Maritimes. Antimicrobial drug use rate was higher via systemic route as compared with intramammary and other routes of administration (topical, oral, and intrauterine). The ADUR of antimicrobials used intramammarily was higher for clinical mastitis treatment than dry cow therapy. For dry cow therapy, penicillin ADUR was greater than ADUR of first-generation cephalosporins. For clinical mastitis treatment, ADUR of intramammary penicillin combinations was greater than ADUR of cephapirin. Herd-level milk production was positively associated with overall ADUR, ADUR of systemically administered ceftiofur, cephapirin administered for dry cow therapy, and pirlimycin administered for clinical mastitis treatment. Herd size and ADUR of systemically administered ceftiofur were also positively associated. In conclusion, β-lactams were most commonly used on Canadian dairy farms. Among antimicrobials of very high importance in human medicine, the use of fluoroquinolones was rare, whereas third-generation cephalosporins and penicillin combinations containing colistin were used very frequently on Canadian dairy farms.

Costs and feasibility of extensive sample collection and processing are major obstacles to mastitis epidemiology research. Studies are often consequentially limited, and fundamental mastitis researchers rarely have the opportunity to conduct their work in epidemiologically valid populations. To mitigate these limitations, the Canadian Bovine Mastitis Research Network has optimized research funds by creating a data collection platform to provide epidemiologically meaningful data for several simultaneous research endeavors. This platform consists of a National Cohort of Dairy Farms (NCDF), Mastitis Laboratory Network, and Mastitis Pathogen Culture Collection. This paper describes the implementation and operation of the NCDF, explains its sampling protocols and data collection, and documents characteristics, strengths and limitations of these data for current and potential users. The NCDF comprises 91 commercial dairy farms in 6 provinces sampled over a 2-yr period. Primarily Holstein-Friesian herds participating in Dairy Herd Improvement milk recording were selected in order to achieve a uniform distribution among 3 strata of bulk tank somatic cell counts and to reflect regional proportions of freestall housing systems. Standardized protocols were implemented for repeated milk samplings on clinical mastitis cases, fresh and randomly selected lactating cows, and cows at dry-off and after calving. Just fewer than 133,000 milk samples were collected. Demographic and production data were recorded at individual cow and farm levels. Health management data are documented and extensive questionnaire data detailing farm management and cleanliness information are also captured. The Laboratory Network represents coordinated regional mastitis bacteriology laboratories using standardized procedures. The Culture Collection archives isolates recovered from intramammary infections of cows in the NCDF and contains over 16,500 isolates, all epidemiologically cross-referenced between linked databases. The NCDF is similar to Canadian dairies in relation to mean herd size, average production, and freestall percentages. Pathogen recovery was greater than anticipated, particularly for coagulase-negative staphylococci and Corynebacterium spp. International scientists are encouraged to use this extensive archive of data and material to enhance their own mastitis research.

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This study was conducted to evaluate the association between subclinical intramammary infection (IMI) with coagulase-negative staphylococci (CNS), mammary quarter milk somatic cell count (SCC), and persistence of IMI in dairy cattle. Convenience samples of CNS isolates harvested from milk samples of subclinically infected mammary quarters collected between 4 and 2wk before drying-off, between 2wk before drying-off and the day of drying-off, within 24h after calving, between 1 and 2wk after calving, and during lactation were evaluated. Isolates were obtained from the Canadian Bovine Mastitis Research Network culture bank and were identified to the species level using rpoB gene sequencing. Cow and quarter-level data were obtained from the Canadian Bovine Mastitis Research Network database and used for statistical analyses. In addition, for mammary quarters that had more than one isolation of the same CNS species at different time points, the isolates were evaluated using pulsed-field gel electrophoresis to identify persistent IMI. Milk SCC was compared between mammary quarters infected with different CNS species and to a cohort of uninfected mammary quarters. A total of 877 isolates from 643 mammary quarters of 555 cows on 89 Canadian dairy farms were identified to the species level. Twenty different species were identified, with Staphylococcus chromogenes being the most common species identified (48% of isolates), followed by Staphylococcus simulans (19%) and Staphylococcus xylosus (10%). Of the 20 species identified, only 9 species were found in persistently infected quarters. Milk SCC was significantly higher in the CNS-infected mammary quarters than in the uninfected control quarters for 8 of the 20 species studied. Also, mean SCC differed significantly between mammary quarters infected with different CNS species. Within a given species, a high degree of variability was noted in milk SCC. These data corroborate recent data from Europe with regard to the predominance of certain species of CNS (e.g., Staph. chromogenes). In addition, some species of CNS appear to have a greater effect on milk SCC. Finally, some CNS species are associated with persistent IMI suggesting that some species (e.g., Staph. chromogenes and Staph. simulans) are better host-adapted, whereas others may have an environmental reservoir.

A systematic review of the scientific literature on relationships between management practices used on dairy farms and herd somatic cell count (SCC) was undertaken to distinguish those management practices that have been consistently shown to be associated with herd SCC from those lacking evidence of association. Relevant literature was identified using a combination of database searches (PubMed, Medline, CAB, Agricola, and Web of Science) and iterative screening of references. To be included in the review, a manuscript had to be published after 1979 in French, English, or Dutch; study design had to be other than case report or case series; herds studied had to be composed of ≥ 40 milking cows producing on average ≥ 7,000kg of milk in 305 d; interventions studied had to be management practices applied at the herd level and used as udder health control strategies; and SCC had to be measured using electronic cell counting methods. The 36 manuscripts selected were mainly observational cross-sectional studies; 8 manuscripts dealt exclusively with automatic milking systems and 4 with management of calves and heifers and its effect on SCC in early lactation heifers. Most practices having consistent associations with SCC were related to milking procedures: wearing gloves during milking, using automatic take-offs, using postmilking teat dipping, milking problem cows last, yearly inspection of the milking system, and use of a technique to keep cows standing following milking; all were consistently associated with lower herd SCC. Other practices associated with lower SCC were the use of a freestall system, sand bedding, cleaning the calving pen after each calving, surveillance of dry-cow udders for mastitis, use of blanket dry-cow therapy, parenteral selenium supplementation, udder hair management, and frequent use of the California Mastitis Test. Regarding SCC of heifers, most of the consistent associations reported were related to interventions made during the peripartum period. Studies on automatic milking systems have frequently reported elevation of the herd SCC following transition to the new system. These elevations seemed to be mediated both by the lack of monitoring of chronically infected cows and by an elevated incidence of intramammary infections. By assembling the results reported in many different studies, this review generates a more comprehensive understanding of the management practices influencing SCC and highlights areas of SCC control knowledge that lack evidence of effectiveness.

Monitoring of antimicrobial resistance (AMR) in bacteria has clinical and public health significance. The present study determined prevalence of AMR in common mastitis pathogens Staphylococcus aureus, including methicillin-resistant Staph. aureus (MRSA; n=1,810), Escherichia coli (n=394), and Klebsiella species (n=139), including extended-spectrum β-lactamase (ESBL)-producing E. coli and Klebsiella species, isolated from milk samples on 89 dairy farms in 6 Canadian provinces. Minimum inhibitory concentrations (MIC) were determined using the Sensititer bovine mastitis plate (Trek Diagnostic Systems Inc., Cleveland, OH) and a National Antimicrobial Resistance Monitoring System gram-negative panel containing antimicrobials commonly used for mastitis treatment and control. Denim blue chromogenic agar and real-time PCR were used to screen and confirm MRSA, respectively. Resistance proportion estimates ranged from 0% for cephalothin and oxacillin to 8.8% for penicillin in Staph. aureus isolates, and 15% of the resistant Staph. aureus isolates were multidrug resistant. One MRSA isolate was confirmed (prevalence: 0.05%). Resistance proportion estimates ranged from 0% for ceftriaxone and ciprofloxacin to 14.8% for tetracycline in E. coli, and 0% for amikacin, ceftiofur, ciprofloxacin, and nalidixic acid to 18.6% for tetracycline in Klebsiella species isolates. Further, 62.8 and 55% of the resistant E. coli and Klebsiella species isolates were multidrug resistant, respectively. Resistance to >5 and >2 antimicrobials was most common in E. coli and Klebsiella species isolates, respectively, and no ESBL producers were found. Prevalence of AMR in bovine mastitis pathogens was low. Most gram-negative udder pathogens were multidrug resistant; MRSA was rarely found, and ESBL E. coli and Klebsiella species isolates were absent in Canadian milk samples.

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.