Surveillance for multidrug resistant Escherichia coli carriage in cattle, dogs and humans reveals predominance of CMY-2, CTX-M-15 and CTX-M-9 groups of ß-lactamases.

Global spread of antimicrobial multidrug resistance (MDR) in human and veterinary medicine relies upon diagnostics, surveillance and stewardship to guide mitigation. Utilizing surveillance of fecal samples from our service area for detecting MDR Escherichia coli carriage in humans (2143), dogs (627), and cattle (130), we found isolates resistant to third/fourth generation cephems present in 3.7 %, 13.1 %, and 51.5 %, respectively. CMY-2, CTX-M-15-like and CTX-M9 group genes in descending order were predominant in all hosts and accounted for 83.3 % of non-wild-type gene targets. MDR carriage mirrored cephem non-susceptibility rates as published in annual antibiograms for humans and dogs; notably, no carbapenem-resistant carriage isolates were detected. Given the scale of MDR E. coli carriage in cattle (14X) and dogs (3.5X) compared to humans, bench-marking of the resistance gene pool by host species utilizing regional One Health surveillance may aid in assessing occupational and geographic risks for acquiring resistance and for monitoring of mitigation strategies.

Multisite reproducibility of the broth microdilution method for susceptibility testing of Nocardia species.

Antimicrobial susceptibility testing (AST) of clinical isolates of Nocardia is recommended to detect resistance to commonly used antimicrobial agents; such testing is complicated by difficulties in inoculum preparation and test interpretation. In this study, six laboratories performed repetitive broth microdilution testing on single strains of Nocardia brasiliensis, Nocardia cyriacigeorgica, Nocardia farcinica, Nocardia nova, and Nocardia wallacei. For each isolate, a total of 30 microdilution panels from three different lots were tested at most sites. The goal of the study was to determine the inter- and intralaboratory reproducibility of susceptibility testing of this group of isolates. Acceptable agreement (>90% agreement at ±1 dilution of the MIC mode) was found for amikacin, ciprofloxacin, clarithromycin, and moxifloxacin. After eliminating MIC values from single laboratories whose results showed the greatest deviation from those of the remaining laboratories, acceptable agreement was also found for amoxicillin-clavulanic acid, linezolid, minocycline, and tobramycin. Results showed unsatisfactory reproducibility of broth microdilution testing of ceftriaxone with N. cyriacigeorgica and N. wallacei, tigecycline with N. brasiliensis and N. cyriacigeorgica, and sulfonamides with N. farcinica and N. wallacei. N. nova ATCC BAA-2227 is proposed as a quality control organism for AST of Nocardia sp., and the use of a disk diffusion test for sulfisoxazole is proposed as a check of the adequacy of the inoculum and to confirm sulfonamide MIC results.

Multilaboratory testing of antifungal combinations against a quality control isolate of Candida krusei.

Candida krusei ATCC 6258 was tested by eight laboratories using 96-well plates containing checkerboard pairwise combinations of amphotericin B (AMB), posaconazole (PSC), caspofungin (CSP), and voriconazole (VRC). The methodology led to reproducible results across the laboratories. All drug combinations yielded MICs lower than the MICs of any two drugs tested singly, and combinations of AMB, PSC, CSP, and VRC were indifferent (no antagonism) by summations of fractional inhibitory concentration.

Factors influencing broth microdilution antimicrobial susceptibility test results for dalbavancin, a new glycopeptide agent.

Performance of antimicrobial susceptibility tests with new agents requires careful consideration of the properties of the antimicrobial to ensure that the tests are standardized, reproducible, and reflect the true potency of the drug. Dalbavancin is a new glycopeptide with potent activity against gram-positive bacterial species. The investigations described here demonstrated that methodologic modifications of procedures are necessary to ensure consistent test results, both for quality control and for routine testing of clinical isolates. Dimethyl sulfoxide is the preferred primary solvent. The addition of 0.002% polysorbate-80 (a surfactant) to dalbavancin-containing wells in the reference broth microdilution assay resulted in consistent and reproducible MIC results for three quality control strains: Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, and Streptococcus pneumoniae ATCC 49619. The same degree of consistency was observed among clinical isolates of gram-positive bacterial species tested in several clinical laboratories. These results indicate that the addition of 0.002% (final concentration) of the surfactant in broth microdilution tests produces optimal dalbavancin MICs required for accurate and reproducible clinical laboratory tests, without untoward influences of substrate binding or media constituents.