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Development of a broth microdilution testing method for aquatic isolates at 22degreeC and 28degreeC using Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29212 as quality control strains


, : Development of a broth microdilution testing method for aquatic isolates at 22degreeC and 28degreeC using Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 29212 as quality control strains. Abstracts of the General Meeting of the American Society for Microbiology 103: C-063

Many bacterial pathogens isolated from aquatic animals require growth conditions that are considerably different from the growth conditions required by bacterial pathogens of warm-blooded animals. To accurately assess the susceptibility of these bacterial pathogens to antimicrobial agents it is necessary to develop appropriate standardized antimicrobial susceptibility testing (AST) methods. To facilitate this, the National Committee for Clinical Laboratory Standards (NCCLS) established an Aquaculture Working Group, charged with developing standardized AST methods for bacterial pathogens of aquatic species. One of the testing methods being developed is a broth microdilution (BMD) method. Following NCCLS guidelines, two ATCC(R) reference strains, E. coli 25922 and S. aureus 29212 have been shown to be excellent quality control (QC) strains for BMD testing at the lower temperatures of 22degreeC and 28degreeC. These organisms have a wide temperature tolerance range and are susceptible to a wide range of antimicrobials commonly used in aquaculture. When the incubation temperature was lowered from the standard QC temperature of 35degreeC, E. coli 25922 and S. aureus 29212, in many cases, demonstrated lower minimum inhibitory concentrations (MICs). This was most likely due to a decrease in the rate of growth caused by the decrease in temperature. In order to evaluate the utility of E. coli 25922 and S. aureus 29212 as control strains, their performance was compared to that of several aquatic isolates tested at 22degreeC or 28degreeC against 10 antimicrobials commonly used in aquaculture. These isolates included Aeromonas spp., Vibrio spp., Edwardsiella spp., Photobacterium spp., and Pseudomonas spp. The results from this study showed that the drug concentration required to inhibit the growth of the two QC organisms was in the same range as the MICs for most of the aquatic isolates tested. The next phase of the study is to demonstrate intra- and inter-laboratory reproducibility in accordance with NCCLS guidelines. The results of these studies will provide a foundation for developing international standardized BMD methods for testing aquatic animal isolates.

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