In vitro bactericidal activity of enrofloxacin against gyrA mutant and qnr-containing Escherichia coli isolates from animals.

The objective of this work was to investigate the bactericidal activity of enrofloxacin against gyrA mutant and qnr-containing Escherichia coli isolates from animals. The minimum inhibitory concentrations (MICs) of gyrA mutant and qnr-containing E coli isolates ranged from 1 µg/ml to 32 µg/ml for enrofloxacin. Time-kill experiments were performed using selected E coli isolates. For the time-kill experiments, the colony counts were determined by plating each diluted sample onto plate count agar and an integrated pharmacokinetic/pharmacodynamics area measure (log ratio area) was applied to the colony-forming units (cfu) data. In general, enrofloxacin exhibited bactericidal activity against all the gyrA mutant E coli isolates at all concentrations greater than four times the MIC. However, the bactericidal activity of enrofloxacin for all the qnr-containing E coli isolates was less dependent on concentration. The results of the present study indicated that the genetic mechanism of resistance might account for the different bactericidal activities of enrofloxacin observed for the gyrA mutant and the qnr-containing E coli isolates. Therefore, in addition to MIC assays, genetic mechanism-based pharmacodynamic models should be used to provide accurate predictions of the effects of drugs on resistant bacteria.

Molecular characterisation of quinolone resistance in Escherichia coli from animals in Turkey.

The aim of this study were to detect the gyrA, parC and marR mutations and qnr genes (qnrA, qnrB and qnrS) in 120 strains of Escherichia coli isolated from animals. European Committee on Antimicrobial Susceptibility Testing and Clinical Laboratory Standards Institute disc diffusion and minimum inhibitory concentration (MIC) tests, respectively, were used to determine fluoroquinolone (FQ) resistance, and molecular methods were used to detect the mutations and the genes. E coli isolates with an MIC of ≥8 mg/l had mutation at Ser-80 in parC in addition to mutations at Ser-83, Asp-87 or both in gyrA. The nucleotide change was detected in marR (Ser-3 → Asn, Ala-53 → Glu, Gly-103 → Ser, Tyr-137 → His). Only four E coli isolates (3.3 per cent) contained qnrA and qnrS, and qnrB was not detected. Two E coli isolates from healthy calves also contained qnrA and qnrS. The MICs of enrofloxacin and danofloxacin for qnr-containing E coli isolates ranged from 32 mg/l to 256 mg/l. The results of this study indicated that the FQ-resistant E coli isolates presented an alteration in gyrA (Ser-83 → Leu, Asp-87 → Asn) and parC (Ser-80 → Ile) with high MICs (8-256 mg/l), and there was a low prevalence of qnr genes among E coli isolated from animals.

Comparison of direct culture versus PCR for the detection of Brucella in aborted fetuses of cattle and sheep in Turkey.

The aim of this study was to detect Brucella in samples from aborted fetuses of sheep and cattle in Turkey using PCR and bacteriological analysis, and to determine the sensitivity and specificity of the PCR. Organ homogenates from 38 aborted fetuses of cattle and 56 aborted fetuses of sheep were tested. All organ homogenates were cultured for bacteriological analysis, and all of the homogenates and the Brucella isolates obtained by culture were examined with a commercial PCR kit. On bacteriological analysis, Brucella species were found in 30 (31.9 per cent) of the 94 organ homogenates, eight (21.1 per cent) of which were from cattle and 22 (39.3 per cent) from sheep. Using PCR, a total of 29 (30.9 per cent) homogenates were positive for Brucella species, eight (21.1 per cent) of which were from cattle and 21 (37.5 per cent) from sheep. Compared with the bacteriological method, the diagnostic sensitivity and specificity of the PCR kit used in this study were 83 per cent and 94 per cent, respectively.

Characterisation of Brucella abortus biovar 3 isolates from Turkey as biovar 3b.

Between 2002 and 2007, 75 strains of Brucella abortus were isolated from aborted bovine fetuses collected from several regions of Turkey. The isolates were all identified as B abortus biovar 3 by conventional methods. However, when they were analysed by enhanced amos-ery pcr, a 5.4 kb deletion, different from that in the Tulya strain (B abortus biovar 3, atcc 23450), was identified in all of them. As a result, they were subtyped as B abortus biovar 3b.