Quinolone Resistance amongSalmonella entericafrom Cattle, Broilers, and Swine in Denmark

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Abstract
This study was conducted to determine the susceptibility to nalidixic acid and fluoroquinolones of Salmonella Dublin, S. Enteritidis, and S. Typhimurium isolates from cattle, broilers, and pigs over time in Denmark and to characterise the gyrA, gyrB, and parC genes in quinolone-resistant isolates. A total of 584 S. Typhimiurium and 573 S. Dublin isolates from cattle during 1984 through 1999, and 241 S. Enteritidis and 131 S. Typhimurium from broilers and 452 S. Typhimurium from pigs isolated during 1997-1999 were tested. All isolates from cattle from the period 1984 through 1992 were susceptible to quinolones. A single (1.1%) S. Typhimurium isolate from 1995 and three (5.9%) from 1998 were resistant to nalidixic acid. Six (9.0%) S. Dublin isolates from 1996, four (4.2%) from 1997, and one (1.7%) from 1998 were resistant to nalidixic acid. Resistance was not observed among isolates from cattle in 1999. All broiler isolates from 1997 except for one were susceptible to nalidixic acid, whereas seven (6.2%) S. Enteritidis and two (6.3%) of the S. Typhimurium isolates from 1998 and 9 S. Enteritidis (26.5%) from 1999 were resistant. Among isolates from pigs, four isolates from 1997, three from 1998, and one from 1999 were resistant to nalidixic acid. All the nalidixic acid-resistant isolates had reduced susceptibility to fluoroquinolones. Sequence analysis of the gyrA gene in 37 nalidixic-resistant isolates identified two different base substitutions at codon serine-83 and two at aspartate-87. The base substitutions in serine-83 were TCC (Ser)-->TAC (Tyr), and TCC (Ser)-->TTC (Phe). The base substitutions in aspartic-87 were GAC (Asp)-->AAC (Asn), and GAC (Asp)-->GGC (Gly). Sequence analysis of the gyrB and parC genes revealed no mutations in 27 selected isolates. This study showed that quinolone-resistant isolates have emerged in recent years among food-producing animals, especially among S. Enteritidis from broilers in Denmark, and that the resistance mainly is associated with mutations in gyrA.