The Dissemination of a Single Staphylococcusaureus Strain, Spa-t2873, as the Predominant Cause of Bovine Mastitis in Israeli Dairy Farms.

Staphylococcus aureus is one of the most important pathogens causing intramammary infection (IMI) in dairy herds. The goals of this study were (1) to describe the prevalence of S. aureus in Israeli dairy farms; (2) to characterize the spa-based clonal structure of mastitis-related S. aureus isolates; (3) to analyze the transmission network of a large outbreak within a single farm and (4) to characterize the virulence factors of the outbreak strain. The prevalence and the molecular survey were performed on all Israeli IMI-related isolates, 9.2019-8.2020. Molecular methods included spa-typing for the survey and whole-genome sequencing (WGS) for the investigation of the farm 'A' outbreak. During the one-year survey, S. aureus was identified in 152 dairy farms, with a total of 440 positive samples. The spa t2873 was found in 284 isolates (64.5%) across 112 farms (73.6%). Other common types included t529 (n = 46), t9303 (n = 34) and the methicillin-resistant S. aureus t011 (n = 11). The highest number of cases (n = 25) was detected in Farm 'A', all of which were found to be spa t2873. Phylogenetic analysis confirmed that most transmission events occurred within the same milking group, and inter-group transmission was due to the transfer of cows between groups or due to consecutive milking order. The spa t2873 strain contained putative virulence genes, including various intracellular and collagen adhesion proteins. Our study revealed the dissemination of the t2873 strain to the majority of Israeli dairy farms. The possibility of inter-farm transmission should be monitored and prevented.

From six to zero per cent Mycobacterium avium subsp. paratuberculosis milk ELISA positivity in three years - probably induced by Mycobacterium vaccae.

This research communication reports the results of a study aimed at investigating the effects of introducing Mycobacterium vaccae on paratuberculosis carriage in a dairy herd. M. vaccae is a non-pathogenic member of the Mycobacteriaceae, with immunomodulatory and immunotherapeutic capabilities, acting by stimulating the cellular immune system, important in protection against paratuberculosis. Starting in 2014 we administered, by gavage, 1010 live M. vaccae bacteria to all new-born heifers on a dairy farm, first within 24 h of birth and again 2 weeks later. Paratuberculosis carriage was monitored yearly by milk ELISA. Faecal samples of 50% of cows, aged 3 years, born 1, 2 or 3 years before the experiment's onset, were tested by qPCR for MAP shedding and compared to 100% treated cows of the same age. Within 3 years, milk ELISA positivity was reduced from 6 to 0% and remained unchanged for the subsequent 2 years. One qPCR positive control cow was found each year for a total of 3 animals (2.46%). One positive cow (1%) was found among the treated cows. Two of the 3 positive control animals, still present on the farm at the end of 2019, tested negative whereas the positive test cow continued shedding MAP. M. vaccae shedding heifers mixing with adult cows were the probable means of the microorganism's propagation. The results of this investigation indicate that the introduction of live M. vaccae may be an inexpensive and fast alternative to current paratuberculosis control practices, justifying further exploration of the topic.

Bovine subclinical mastitis caused by Mannheimia granulomatis.

Mannheimia granulomatis was isolated for 10 months from the milk of a cow with elevated somatic cell counts. The infection was self-limiting. Phenotypic and molecular characteristics of the isolate were determined.

Assessment of accuracy of disk diffusion tests for the determination of antimicrobial susceptibility of common bovine mastitis pathogens: a novel approach.

A novel approach was used to assess disk diffusion accuracy for determination of antibiotic susceptibility of various bovine mastitis pathogens (Escherichia coli, Staphylococcus aureus, Staphylococcus chromogenes, and Streptococcus dysgalactiae). MIC and disk diffusion diameters were compared for 587 bovine mastitis bacterial isolates collected in Israel and 3,186 drug-organism combinations. Results were analyzed by ROC curves, Bayesian statistics, and standard descriptive methods. Low correlation was observed between results of disk diffusion and MIC for S. dysgalactiae and all antimicrobial agents, S. aureus and erythromycin and neomycin, and E. coli and gentamicin, neomycin, and polymyxin B. On a few occasions in which correlation was satisfactory, accepted susceptibility breakpoints to some of the antimicrobial agents resulted in high discrepancies with MIC results and new breakpoints were suggested-e.g., 21 mm for S. aureus susceptibility to penicillin G instead of 29 mm recommended by the National Committee for Clinical Laboratory Standards (NCCLS) and <21 mm, resistant, 21-25 mm, intermediate, and >25 mm, susceptible for susceptibility of E. coli to trimethoprim/sulfamethoxazole. Thus, this approach enabled determination of the most accurate breakpoints that best fitted the specific prevalence of susceptibility in Israel. Thus, we suggest its adoption by microbiology diagnostic laboratories for the provision of accurate antimicrobial susceptibility results when using the disk diffusion test.