Detection of efflux pump CmeABC in enrofloxacin resistant Campylobacter spp. strains isolated from broiler chickens (Gallus gallus domesticus) in the state of Rio de Janeiro, Brazil

Abstract

Fowls are the main reservoirs of the highly important food-originating pathogen called Campylobacter spp. and broilers’ meat and byproducts are the main vehicles of this microorganism. Increasing of Campylobacter spp. resistant strains to fluorquinolones, an antimicrobial class often employed in poultry farming and in human medicine has become a great concern to poultry breeders. In fact, several studies evaluated increasing bacterial resistance against these antimicrobial agents. The role of CmeABC efflux system has been underscored among the resistance mechanisms in Campylobacter spp. to fluorquinolones. This study investigated the occurrence of CmeABC efflux pump in 81 and 78 enrofloxacin resistant strains of Campylobacter jejuni and C. coli respectively, isolated from broilers collected from six abattoirs situated at São José do Vale do Rio Preto/RJ poultry center and from two commercial abattoirs situated at Metropolitan Region of Rio de Janeiro, from 2013 to 2016. The resistance to enrofloxacin was assessed by agar dilution to determine minimum inhibitory concentration (MIC). The CmeABC efflux system was investigated through the detection of genes genes cmeA, cmeB and cmeC by PCR. The activity of CmeABC efflux pump was investigated in 20 strains by using the efflux pump inhibitor Phenylalanine-Arginine β-Naphthylamide (PAβN). The three genes cmeA, cmeB and cmeC were detected in 94.3% of the strains (C. jejuni = 80 and C. coli = 70), whereas the system was absent or incomplete in 5.7% of strains (C. jejuni = 1 and C. coli = 8). MIC varied between 0.5μg/ml and 64μg/ml, and 88.7% of strains were enrofloxacin resistant and 11.3% featuring intermediate resistance. The inhibition of the efflux pump by PAβN reduced the MIC to enrofloxacin up to eight times in fifteen strains (75%). These results indicate that this system is frequent and active in Campylobacter spp. Resistant strains in the presence of enrofloxacin.

Antimicrobial Resistance Challenged with Platinum(II) and Palladium(II) Complexes Containing 1,10-Phenanthroline and 5-Amino-1,3,4-Thiadiazole-2(3H)-Thione in Campylobacter jejuni

“…The presence of the ermB gene encoding 23S rRNA methyltransferase and the cmeABC multidrug efflux pump has also been shown to be involved in acquired resistance to erythromycin [ 41 ]. The latter has been identified in 80/235 (34.0%) strains in our study to date, including susceptible strains [ 40 ].…”

“…The latter has been identified in 80/235 (34.0%) strains in our study to date, including susceptible strains [ 40 ]. The presence of this efflux pump is favorable to the pathogen, not only in the elimination of antimicrobials but also in the excretion of metabolites and bile salts harmful to C. jejuni [ 39 , 41 ].…”

Antimicrobial Resistance in Campylobacter jejuni Isolated from Brazilian Poultry Slaughterhouses

“…Mutation in QRDR of gene gyrA is the main resistance mechanism to fluoroquinolones. However, chromosomal efflux pumps, especially those codified by cmeA, cmeB and cmeC genes, are important factors to antimicrobial in Campylobacter species (Wieczorek & Osek, 2013;Nascimento et al, 2019). Previous studies have demonstrated that almost all strains of Campylobacter jejuni isolated in Brazil presented these three genes (Nascimento et al, 2019).…”