Prevalence and characterization of plasmid-mediated quinolone resistance genes in Salmonella isolated from poultry in Korea.

The purpose of this study was to investigate the prevalence and characteristics of plasmid-mediated quinolone resistance (PMQR) genes qnr, aac(6')-Ib-cr, and qepA in a total of 185 non-duplicate Salmonella spp. isolated from hatcheries, poultry farms, and poultry slaughterhouses during the period 2001 to 2010 in Korea. Additionally, mutation analysis of quinolone resistance determining regions (QRDRs), conjugation experiments, and plasmid analysis were performed in the PMQR-positive isolates. Among the 185 isolates, six (3.2%) contained qnr genes (two qnrB4 and four qnrS1) but none carried the aac(6')-Ib-cr or qepA genes. Among the six PMQR-positive isolates, one showed a single mutation (Ser83-Phe substitution) in the QRDRs of gyrA. Among them, three were non-susceptible (intermediate or resistant) to nalidixic acid (minimum inhibitory concentration [MIC] ≥256 µg/ml), ciprofloxacin (MIC 2 µg/ml), and levofloxacin (MIC 4 µg/ml), but others were susceptible to all of the three fluoroquinolones. They were resistant to six or more antimicrobial agents tested and were able to transfer quinolone resistance to recipient Escherichia coli J53 by conjugation. By performing a hybridization test, plasmids harbouring qnrB4 and qnrS1 genes were less than 8 kb and about 70 kb in size, respectively. The horizontal dissemination of qnrS1 gene was mediated by IncN plasmid. Compared with the recipient strain, MICs of the transconjugants increased two-fold to four-fold for nalidixic acid, and eight-fold to 16-fold for ciprofloxacin and levofloxacin. This report is the first to describe the detection of qnr genes in Salmonella spp. isolated from poultry in Korea. Widespread horizontal transfer of these genes among bacteria may be a serious public health concern because these can rapidly increase fluoroquinolone resistance. To ensure the public health, it is essential to continuously survey and carefully monitor the spread of PMQR genes in Salmonella from poultry.

Characterization of Salmonella spp. isolated from an integrated broiler chicken operation in Korea.

The purpose of this study was to investigate the biological and genetic characterization of persistent Salmonella isolates in an integrated broiler chicken operation, in an attempt to elucidate the source of contamination. From the breeder farm, the hatchery, the broiler farm and the chicken slaughter house of an integrated broiler chicken operation, a total of 6 serotypes were observed. Although S. Heidelberg was not detected in the broiler farm, it was consistently found in the breeder farm, the hatchery and the chicken slaughter house. Also, S. Enteritidis and S. Senftenberg were found in the hatchery and the chicken slaughter house, and the hatchery and the broiler farm, respectively. S. Gallinarum and S. Blockley were found only in the broiler farm, and S. Virchow was only recovered in the chicken slaughter house. Isolated S. Heidelberg, S. Enteritidis and S. Senftenberg strains were divided into 3, 5 and 7 types, respectively, on the basis of all properties. Especially, S. Senftenberg isolates, divided into four types by their antimicrobial resistance patterns, were all obviously the XbaI PFGE pattern. Also, four S. Enteritidis isolates resistant to nalidixic acid showed a difference in phage type and PFGE pattern. Such a different pattern was shown despite Salmonella isolates originating from an integrated broiler operation, suggesting that further epidemiological studies on many integrated chicken companies in Korea are needed.

Fluoroquinolone resistance and gyrA and parC mutations of Escherichia coli isolated from chicken.

Escherichia coli is a common inhabitant of the intestinal tracts of animals and humans. The intestines of animals also represent an ideal environment for the selection and transfer of antimicrobial resistance genes. The aim of this study was to investigate the resistance of E. coli isolated from chicken fecal samples to fluoroquinolones and to analyze the characterization of mutations in its gyrA and parC gene related resistance. One hundred and twenty-eight E. coil isolates showed a high resistance to ciprofloxacin (CIP; 60.2%), enrofloxacin (ENO; 73.4%) and norfloxacin (NOR; 60.2%). Missense mutation in gyrA was only found in the amino acid codons of Ser-83 or Asp-87. A high percentage of isolates (60.2%) showed mutations at both amino acid codons. Missense mutation in parC was found in the amino acid codon of Ser-80 or Glu-84, and seven isolates showed mutations at both amino acid codons. Isolates with a single mutation in gyrA showed minimal inhibitory concentrations (MIC) for CIP (

Biochemical characteristics and antimicrobials susceptibility of Salmonella gallinarum isolated in Korea.

Fowl typhoid (FT) reported since 1992 in Korea is a septicemic disease of domestic birds caused by Salmonella gallinarum (S. gallinarum). The purpose of this study was to investigate the biochemical characteristics and antimicrobials susceptibility of field isolates of S. gallinarum isolated by year in Korea. A total of 258 isolates of S. gallinarum from 1995 to 2001 showed the same pattern in the majority of biochemical test such as IMViC (indole, methyl red, Voges-Proskauer and citrate utilization), carbohydrate fermentation and amino acid decarboxylation, and these results were almost in accordance with the traditional biochemical characteristics of S. gallinarum strain. When the antimicrobial susceptibility test against 258 isolates of S. gallinarum was performed by the disk diffusion method using 12 antimicrobial agents, all isolates from 1995 appeared to be susceptible to all of the antimicrobial agents tested except for tetracycline and oxytetracycline, whereas the vast majority of isolates from 2001 showed the reduced susceptibility to ampicillin (13.0%), gentamicin (43.4%), kanamycin (69.6%), enrofloxacin (6.5%), ciprofloxacin (10.9%), norfloxacin (52.5%) and ofloxacin (82. 6%). The prevalence of the prevalence of completely resistant isolates resistant isolates to one or more drugs rapidly increased from 0% in 1995 to 93.5% in 2001. The minimal concentrations range of the majority of antimicrobial agents to inhibit 50% (MIC50s) against S. gallinarum isolates increased from 0.06 approximately 8 microg/ml in 1997 to 2 approximately 256 microg/ml in 2001. Especially, MIC50s for gentamicin and fluoroquinolones of isolates from 2001 increased over 10-fold than those of isolates from 1997. Therefore, our results indicate that sorbitol fermentation and arginine decarboxylation showed the diversity by isolates and the vast majority of isolates from 2001 showed the reduced susceptibility to antimicrobials tested.