β-Lactamase-Mediated β-Lactam Resistance in Campylobacter Species: Prevalence of Cj0299 ( bla _OXA-61 ) and Evidence for a Novel β-Lactamase in C . jejuni

Abstract

Fifty-two percent of 1,288 poultry isolates of campylobacters were ampicillin resistant, and resistance was more common among Campylobacter coli isolates (67.4%) than among Campylobacter jejuni isolates (47.5%). Production of ␤-lactamase was typically associated with resistance to ampicillin, amoxicillin (amoxicilline), penicillin, and ticarcillin. Regardless of ␤-lactamase production, all isolates were resistant to piperacillin (MICs > 256 g/ml), and most were resistant to carbenicillin, cloxacillin, and cephalosporins. Of all ampicillin-resistant campylobacters tested, 91% (347/380) carried the bla OXA-61 gene, and 77% (136/175) of those tested with nitrocefin produced a ␤-lactamase, presumably OXA-61. The isoelectric point (pI) of OXA-61 was 8.7, and the molecular mass was 31.0 kDa. Insertional inactivation of bla OXA-61 in C. jejuni NCTC 11168 and two ampicillin-resistant isolates resulted in increased susceptibility to ampicillin, co-amoxiclav (amoxicillin and clavulanic acid), penicillin, carbenicillin, oxacillin, and piperacillin, but the effects on MICs of cephalosporins and imipenem were negligible. Some C. jejuni isolates that lacked bla OXA-61 produced a ␤-lactamase, CjBla2, with a pI of 9.2 and molecular mass of 32.4 kDa. Mass spectrometry confirmed that the most prevalent ␤-lactamase was the product of bla OXA-61 , but CjBla2 was not identified. OXA-61 is prevalent among Campylobacter spp. of veterinary origin and is similar to the ␤-lactamase previously reported in human isolates. Production of OXA-61 was associated with resistance to penams but not cephalosporins. Co-amoxiclav remained active against all isolates tested.

Resistance to β-lactam and tetracycline in Campylobacter spp.isolated from broiler slaughterhouses in southern Brazil

“…Recently, it was proposed that an oral β-lactam may provide an alternative therapy, although epidemiological studies have shown β-lactam resistance to be commonly observed in Campylobacter spp. (Griggs et al 2009, Gormley et al 2010). β-lactam antimicrobials, which inhibit bacterial cell wall biosynthesis, have been the most commercially available antimicrobials in the market because of their high specificity and potent killing effect (Zeng et al 2014).…”

Genome-wide association study identifies vitamin B 5 biosynthesis as a host specificity factor in Campylobacter

“…S3), with only seven polymorphic sites among 774 nucleotides. This gene, which encodes a characterized enzyme called bla OXA-61 conferring resistance to β-lactam antibiotics (12), was found at the highest frequency in cattle isolates and was rarest in isolates from wild birds (Fig. S1).…”

“…Three upstream genes (surE, Cj0294, and Cj0295) are also involved in biosynthesis. Immediately downstream is an ampicillin resistance-associated gene, bla OXA-61 (12), which is also associated with cattle. The gene is notable for being the least diverse of the common genes in the Campylobacter genome (Fig.…”

Functional Cloning and Characterization of Antibiotic Resistance Genes from the Chicken Gut Microbiome

“…To address this issue, we first constructed an E. coliCampylobacter shuttle vector (pZW) ( Table 1) carrying a Campylobacter promoter according to the procedure described by Larsen et al (8). The two selected novel AR gene products (FRAmp1.1, conferring ampicillin resistance, and FRSpe1.1, conferring spectinomycin resistance) shared low sequence similarity to the major determinants of ampicillin resistance (Cj0299) and spectinomycin resistance (AadA) in Campylobacter (6,17). FRAmp1.1 and FRSpe1.1 were PCR amplified using proper primers pairs (Table 2), and the PCR fragments were cloned into pZW, creating pZW1 and pZW2, respectively.…”