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This study aimed to identify the pathogens isolated from the milk of cows with clinical mastitis in the subtropical region of Australia and to determine the antimicrobial susceptibility of these bacteria. Thirty dairy herds in the subtropical dairy region were asked to submit milk samples for the first 5 cases of clinical mastitis each month for 12 mo. Samples underwent aerobic culture, and isolates were identified via MALDI-TOF mass spectrometry. Antimicrobial susceptibility was determined for Escherichia coli, Enterococcus spp., Streptococcus agalactiae, Streptococcus uberis, Streptococcus dysgalactiae, Staphylococcus aureus, and non-aureus staphylococci and mammaliicocci (NASM). Between March 2021 and July 2022, 1,230 milk samples were collected. A positive culture result was recorded for 812 (66%) of the milk samples; from these samples, 909 isolates were obtained, including 49 isolates where no identification was possible. The remaining samples were classified as having no growth (16.8%) or as being contaminated (17.2%). The most common isolates with a MALDI-TOF diagnosis (n = 909) were Strep. uberis (23.6%), followed by the NASM group (15.0%). Farms enrolled in the study were in 3 distinct locations within the subtropical dairy region: North Queensland, Southeast Queensland, and Northern New South Wales. Some variation in isolate prevalence occurred between these 3 locations. We found lower odds of a sample being positive for E. coli in North Queensland (odds ratio [OR]: 0.25; 95% confidence interval [CI]: 0.07-0.87) and higher odds in Southeast Queensland (OR: 4.01; 95% CI: 1.96-8.20) compared with the reference, Northern New South Wales. We further found higher odds of Strep. dysgalactiae in North Queensland (OR: 5.69; 95% CI: 1.85-17.54) and Southeast Queensland compared with Northern New South Wales (OR: 3.99; 95% CI: 1.73-9.22). Although some seasonal patterns were observed, season was not significant for any of the analyzed isolates. Farm-level differences in pathogen profiles were obvious. Overall, clinical mastitis pathogens had low levels of resistance to the antimicrobials tested. This research demonstrates that Strep. uberis and the NASM bacterial group are the most common pathogens causing clinical mastitis in the subtropical dairy region. It highlights the importance of understanding pathogenic causes of mastitis at the farm and regional level for targeted control and therapy., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

A 5-year retrospective study was conducted to describe the mastitis-causing organisms isolated from bovine milk samples submitted to four veterinary diagnostic laboratories in Australia. The aim of this study was to identify temporal, geographical, and seasonal patterns of occurrence for the organisms and report the in vitro susceptibility of the most common mastitis-causing pathogens. In total, 22,102 milk samples were submitted between 2015 and 2019. The results were reported as positive growth for at least one significant organism ( n = 11,407; 51.6%), no growth ( n = 5,782; 26.2%), and mixed/contaminated growth ( n = 4,913; 22.2%). Culture results for no growth, gram-negative bacteria, and eukaryotic organisms were combined for each region, and they were accounted for between 23 and 46% of submissions. These results represent a subset of mastitis cases for which the antibiotic treatment may not be warranted. A total of 11,907 isolates were cultured from 11,407 milk samples. The most common isolated organisms were Streptococcus uberis [41.3%; 95% confidence interval (CI): 40.4-42.1%] and Staphylococcus aureus (23.6%; 95% CI: 22.8-24.3%). For S. uberis and S. aureus , there was an association between a positive culture result and the dairy region. All regions except for the Sub-tropical Dairy region were more likely to culture S. uberis compared to the reference, Dairy NSW ( P < 0.001). Similarly, for S. aureus , a positive culture result was more likely in all other dairy regions compared to Dairy NSW ( P < 0.001). The LISA cluster analysis identified differences between High-High (hotspot) postcodes for S . aureus and S . uberis throughout all the analyzed dairy regions. These results highlight the need for further investigations into specific risk factors, such as environmental factors and herd-level predictors, which may have influenced the observed regional variations. Common mastitis-causing pathogens showed overall good susceptibility to a range of antimicrobials used in the treatment of mastitis. On-going surveillance of mastitis-causing pathogens and their antimicrobial susceptibilities will facilitate targeted mastitis control and treatment programs., Competing Interests: MH is employed by The Milk Road. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Langhorne, Gupta, Horsman, Wood, Wood, Barker, Deutscher, Price, McGowan, Humphris, Ranjbar, Henning and Gibson.)