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R.K. Cooper

Publications and source records attributed to R.K. Cooper.

5 recordsLinked to original sources
USGS1998

Biochemical and conjugation studies of romet-resistant strains of Aeromonas salmonicida from salmonid rearing facilities in the eastern United States

Strains of Aeromonas salmonicida ( n = 585) were collected from covertly infected and diseased salmonid hosts from 12 hatcheries in the eastern United States. Strains and sites were selected because of their potential for harboring antimicrobial resistance, in particular, to Romet™. Resistance to Romet was displayed by 315 strains (53.8%), which were isolated from all six host species sampled at 10 of 12 sites. Thirty of the resistant strains (9.5%) from five sites had no zone of inhibition, whereas the other strains had either confluent growth or resistant colonies within a zone of inhibition. Fifty-one resistant strains, representing each of the three resistance phenotypes, were selected for biochemical and antimicrobial comparisons with Romet-sensitive strains. All were confirmed to be A. salmonicida , and no characteristic biochemical phenotypes were found to be associated with resistance to Romet. Differential resistances between resistant and sensitive strains were detected to the antimicrobials oxytetracycline, tetracycline, sulfadiazine, sulfamethizole, trimethoprim, and SXT, a potentiated sulfonamide composed of trimethoprim and sulfamethoxazole. Plasmid DNA isolation and agarose gel electrophoresis were done for 25 Romet-resistant strains, and R-plasmids, not present in sensitive strains, were detected in 23 of these. Two different sizes of R-plasmids were detected, one about 55 kilobase pairs long and another about 50 kilobase pairs. Two strains isolated from New York brook trout Salvelinus fontinalis had reduced confluent growth within a zone of inhibition but contained no large plasmids. This may indicate chromosomally mediated resistance. Conjugational mating studies evaluated transfer of the R-plasmid DNA using eight Escherichia coli recipients. Successful R-plasmid transfer was accomplished with two donor strains (MI1 and MI2 from New Hampshire brook trout). Our results, in addition to those of other workers, illustrate the widespread resistance in A. salmonicida to approved antimicrobials and the capacity of this bacterium to become resistant in the fish culture environment.

Journal of Aquatic Animal Health↗
USGS1993

Plasmid-mediated romet resistance of Edwardsiella ictaluri

Romet®, a potentiated sulfa drug composed of five parts sulfadimethoxine and one part ormetoprim, is used to treat channel catfish Ictalurus punctatus infected with Edwardsiella ictaluri , the causal agent of enteric septicemia of catfish (ESC). Recently, several Romet-resistant isolates of E. ictaluri were isolated from channel catfish that had died from ESC in Virginia and Mississippi. This antimicrobial resistance was determined to be plasmid-encoded, as shown by agarose gel electrophoresis of plasmid DNA, and by the fact that the plasmid and associated antimicrobial resistance could be transferred to a recipient by single-step conjugation. Size of the R plasmid was approximately 55 kilobase pairs. Plasmids also conferred resistance to tetracycline, oxytetracycline, streptomycin, trimethoprim, and SXT (another potentiated sulfonamide, composed of trimethoprim and sulfamethoxazole). Each Romet-resistant E. ictaluri isolate was mated with a plasmid recipient, the Romet-sensitive Escherichia coli isolate 1932. Transconjugates were selected by plating on Mueller-Hinton agar that contained antimicrobials, and the mean transfer frequency was 1.483 × 10 −3 transconjugates per donor cell in the mating mixture. Each E. coli transconjugate was then mated with 20 Romet-sensitive isolates of E. ictaluri to assess the propensity of these E. ictaluri isolates to accept the R plasmid. The average transfer frequency when E. ictaluri isolates of channel catfish origin (N = 14) were used as recipients was 1.197 × 10 −2 transconjugates per donor cell in the mating mixture; mean transfer frequency when E. ictaluri isolates not originating from channel catfish were used was 1.019 × 10 -3 transconjugates per donor cell, which was significantly less (P = 0.0002).

Journal of Aquatic Animal Health↗
USGS1993

Comparison of plasmids isolated from Romet-30-resistant Edwardsiella ictaluri and tribrissen-resistant Escherichia coli

Edwardsiella ictaluri , the etiological agent of enteric septicemia of channel catfish (ESC) is the leading cause of bacterial disease in commercially raised channel catfish Ictalurus punctatus . The only drug approved by U.S. Food and Drug Administration for use against ESC is Romet-30. Recently, several isolates were obtained that had a naturally occurring resistance to Romet-30. On further characterization these isolates were shown to possess a 55-kilobase (kb) plasmid that encodes resistance to the drug. We compared Romet-30-resistant E. ictaluri and a Tribrissen-resistant strain of Escherichia coli (strain 1898) isolated from equine cystitis. Antimicrobial profiles, plasmid screening, restriction digest, and Southern blot analysis indicated that the two plasmids are very similar. The resistance afforded to the E. coli was encoded by a 55-kb plasmid. Each of the R plasmids conferred resistance to Romet-30, tetracycline, and Terramycin (oxytetracycline). The R plasmid from E. coli strain 1898 was transferred to Romet-30-sensitive isolates of E. ictaluri by single-step conjugation, rendering the transconjugates resistant to Romet30, tetracycline, and Terramycin. Each plasmid was cleaved into fragments by restriction enzymes, then electrophoresed in an agarose gel and transferred to nitrocellulose. A labeled probe prepared from the R plasmid of E. ictaluri isolate 5-90-156, one of the original Romet-30-resistant E. ictaluri isolates, was added. Restriction enzyme digestion resulted in an equal number of fragments of equal mobility for the two plasmids, and the probe hybridized with the same fragments of each plasmid. These results indicate that the R plasmids of E. ictaluri and E. coli are similar, if not identical.

Journal of Aquatic Animal Health↗