High‐temperature short‐time (HTST) pasteurisation is not sufficient for complete elimination of Bacillus sp. in milk. Hence, it is important to assess their prevalence, phenotypic and genetic antimicrobial resistance profiles in pasteurised milk. In this study, we investigated Bacillus prevalence and their phenotypic resistance to 25 antimicrobials and prevalence of five plasmid antimicrobial resistance genes. All isolates were resistant to β‐lactams; most isolates were susceptible to chloramphenicol ( n = 52), ciprofloxacin, meropenem, sulphonamides, tetracycline and vancomycin ( n = 56). Intriguingly, despite phenotypic sulphonamide and tetracycline sensitivity, Sul2 and tetA genes were detected in different Bacillus species, signifying a potential risk of horizontal gene transfer of these mobile resistance genes through pasteurised milk.
The emergence of macrolide and tetracycline resistance within Pasteurella multocida isolated from feedlot cattle and the dominance of ST394 in Australia was reported recently.
Abstract Background Biofilm production by canine otitis externa (COE) pathogens and resistance development to multiple antimicrobials are commonly reported problems in veterinary practice. The use of adjuvants to disrupt biofilms may be a viable adjunctive therapy. Hypothesis/Objectives To compare the in vitro antibiofilm activity against COE pathogens of three otic cleansers: PHMB‐EDTA (poly [hexamethylene] biguanide hydrochloride and disodium edetate), N ‐acetylcysteine (NAC) and Triz‐EDTA. Animals/Isolates Thirty isolates of each species, including Staphylococcus pseudintermedius, Pseudomonas aeruginosa, Streptococcus canis, Proteus mirabilis, Escherichia coli , and Malassezia pachydermatis , were collected from COE cases and stored at −80°C until tested. Methods and Materials Biofilm production was determined by Congo‐red agar and microtitre plate‐assay methods. Ten of the best biofilm‐producing isolates per species were selected to determine minimum biofilm eradication concentration (MBEC) values. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were determined to compare MBEC/MIC and MBEC/MBC. Results PHMB‐EDTA possessed antibiofilm activity at low concentrations (MBEC range 3.9/2.3–500/300 μg/mL) against all tested COE pathogens. NAC demonstrated antibiofilm activity for all tested bacterial COE pathogens (MBEC range 4,925–19,700 μg/mL); however, most M. pachydermatis isolates exhibited MBEC values >20,000 μg/mL. Triz/EDTA at the highest concentration tested (3,025/19,520 μg/mL) did not demonstrate antibiofilm activity against most COE pathogens except for S. canis (94.5/610 μg/mL). Conclusions and Clinical Relevance PHMB‐EDTA had intrinsic antibiofilm activity at low concentrations against all COE pathogens. Therefore, it is likely to be a very effective adjuvant when used in conjunction with other antimicrobials for the treatment of COE caused by biofilm‐producing pathogens.
The similarity of commensal Escherichia coli isolated from healthy cattle to antimicrobial-resistant bacteria causing extraintestinal infections in humans is not fully understood. In this study, we used a bioinformatics approach based on whole genome sequencing data to determine the genetic characteristics and phylogenetic relationships among faecal Escherichia coli isolates from beef cattle (n = 37) from a single feedlot in comparison to previously analysed pig faecal (n = 45), poultry extraintestinal (n = 19), and human extraintestinal E. coli isolates (n = 40) from three previous Australian studies. Most beef cattle and pig isolates belonged to E. coli phylogroups A and B1, whereas most avian and human isolates belonged to B2 and D, although a single human extraintestinal isolate belonged to phylogenetic group A and sequence type (ST) 10. The most common E. coli sequence types (STs) included ST10 for beef cattle, ST361 for pig, ST117 for poultry, and ST73 for human isolates. Extended-spectrum and AmpC β-lactamase genes were identified in seven out of thirty-seven (18.9%) beef cattle isolates. The most common plasmid replicons identified were IncFIB (AP001918), followed by IncFII, Col156, and IncX1. The results confirm that feedlot cattle isolates examined in this study represent a reduced risk to human and environmental health with regard to being a source of antimicrobial-resistant E. coli of clinical importance.
This study investigated the antimicrobial resistance (AMR) profile of fecal Escherichia coli isolates from beef cattle (n = 150) at entry and exit from an Australian feedlot. Sample plating on MacConkey agar and Brilliance ESBL agar differentiated generic from extended-spectrum β-lactamase (ESBL)-producing E. coli, respectively. Resistance profiles were determined by minimum inhibitory concentration (MIC) testing and further analyzed by whole-genome sequencing (WGS). At entry, the prevalence of antimicrobial resistance to amoxicillin/clavulanic acid, ampicillin, streptomycin, and trimethoprim/sulfamethoxazole was very low (0.7%, each). At the exit, the resistance prevalence was moderate to tetracycline (17.8%) and low to ampicillin (5.4%), streptomycin (4.7%), and sulfisoxazole (3.9%). The most common AMR genes observed in phenotypically resistant isolates were tet(B) (43.2%), aph(3″)-Ib and aph(6)-Id (32.4%), blaTEM-1B, and sul2 (24.3%, each), which are responsible for resistance to tetracyclines, aminoglycosides, β-lactams, and sulfonamides, respectively. The ESBL-producing E. coli were recovered from one sample (0.7%) obtained at entry and six samples (4.0%) at the exit. The ESBL-producing E. coli harbored blaTEM (29.7%), blaCTX m(13.5%), and blaCMY (5.4%). The resistance phenotypes were highly correlated with resistance genotypes (r ≥ 0.85: p < 0.05). This study demonstrated that E. coli isolated from feedlot beef cattle can harbour AMR genes, but the low incidence of medically important resistance reflected the prudent antimicrobial use in the Australian industry.
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