Characterization of Erythromycin-Resistant Porcine Isolates ofCampylobacter coli

Abstract

Erythromycin-resistant Campylobacter organisms were isolated from swine, and their resistance to the antibiotic was characterized. One hundred fourteen Campylobacter organisms were isolated from 572 swine intestinal samples. All isolates were identified as Campylobacter coli by sequence analysis of 16S rRNA gene and polymerase chain reactions with primers specific to hippuricase gene in Campylobacter jejuni and aspartokinase gene in C. coli. Minimal inhibitory concentrations (MICs) of erythromycin were determined by using the agar dilution method, and 80 isolates were found to be resistant to erythromycin (MIC ≥ 4 μg/ml). Of these, 31 isolates had low-level resistance (MIC =4-16 μg/ml), and 49 isolates had high-level resistance (HLR, MIC ≥ 32 μg/ml). The HLR isolates carried a point mutation at position A2075 → G in domain V of the 23S rRNA gene, whereas the low-level resistance isolates carried no mutation. These 49 HLR isolates were characterized by pulsed-field gel electrophoresis and multilocus sequence typing to study their genetic diversity. Pulsed-field gel electrophoresis identified 16 distinct types with 50% genetic similarity as the cutoff. On the other hand, 28 different sequence types (STs), including 10 new STs, were identified with multilocus sequence typing. Forty-six of 49 erythromycin HLR isolates showed crossresistance to 6 macrolide derivatives. The correlation between the inhibitory activity of carbonyl cyanide m-chlorophenylhydrazone and the existence of cmeB, which is responsible for efflux in HLR isolates, was found to be low. Erythromycin resistance was transferred from 38 of the 43 HLR isolates to susceptible C. coli by natural transformation, with a frequency of 1.217 x 10(-8)-4.618 x 10(-5) per recipient cell. All transformants were erythromycin resistant and had A2075 → G mutation in at least one of three copies of the 23S rRNA gene. Results indicate that variable genotypes of HLR C. coli coexist in swine and high-level erythromycin resistance can be transferred to other strains.