Occurrence of aminoglycoside phosphotransferase subclass I and II structural genes among Enterobacteriaceae spp. isolated from meat samples

Abstract

3'-Aminoglycoside phosphotransferase [APH(3')] enzymes are a group responsible for resistance to the antibiotics kanamycin (Km) and neomycin (Nm) in bacteria. Escherichia coli ECT24, originally isolated from a meat sample, harboured an 83-kb conjugative R-plasmid (pRPJ24) that carries transferable resistance to Km and Nm. Plasmid pRPJ24 was transferred by conjugation to Enterobacter cloacae 94R, which was used as the source of plasmid DNA in development of a probe for the Km-resistance determinant. Random cloning of BamHI and HindIII double-digest restriction fragments of pRPJ24 in the pUC18 vector plasmid produced clones resistant to both Nm and Km carrying a 1.9-kb DNA insert. Southern hybridization of pRPJ24 cloned chimeric plasmid DNA (pKPJ94) showed homology with the APH(3')II gene from transposon Tn5. A PstI digest of pKPJ94 produced a 920-bp fragment which hybridized with the APH(3')II structural gene, and was used as a DNA probe for the APH(3')II subclass gene. A 980-bp BamHI fragment from plasmid pGH54 carrying the APH(3')I gene from transposon Tn903 was used as a subclass I probe. Total DNA from 206 randomly screened Km-resistant Enterobacteriaceae isolates from raw ground beef and chicken meat samples were examined for the occurrence of APH(3') subclass I and II using non-radioactively-labelled DNA probes. Thirty-six percent and 60% of the isolates examined carried subclass I and II resistances, respectively, in the isolates from chicken meat samples. The corresponding values for bacterial strains from raw ground beef samples were 51% and 72%, respectively. Four percent of the resistant bacterial isolates from chicken samples did not display homology to either probe.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibiotic Resistance inEscherichia coliIsolated from Retail Raw Chicken Meat in Taif, Saudi Arabia

“…This association has also been demonstrated for antimicrobial agents used for growth promotion (Bager et al, 1997;Aarestrup and Carstensen, 1998;Aarestrup et al, 2000). At slaughter, resistant strains from the gut readily contaminate poultry carcasses, and as a result, poultry meats are often contaminated with resistant E. coli (Linton et al, 1977;Caudry and Stanisich, 1979;Nazer, 1980;Bensink and Botham, 1983;Chaslus Dancla and Lafont, 1985;Jayaratne et al, 1990;Turtura et al, 1990; Van den Bogaard et al, 2001;Vaidya et al, 2005;Akond et al, 2009;Altekruse et al, 2009). The high frequency of antibiotic resistance obtained in the present work is in agreement with that stated by Van et al (2007Van et al ( , 2008.…”

Real-Time Polymerase Chain Reaction Method for Detecting Contamination of Beef by Material from Genetically Engineered Cattle

“…3) We applied this detection method to testing five samples of beef imported from the U.S.A., and trace amounts of the neomycin-resistance gene were detected in one of the five samples in our survey. Sequences homologous to the 3Ј-aminoglycoside phosphotransferase gene, which is responsible for neomycin resistance, have been detected in raw ground beef, 11) consistent with our own results. Since it is almost impossible to completely remove contaminating bacteria containing DNA sequences homologous to the 3Ј-aminoglycoside phosphotransferase gene from beef samples, we considered it important to simultaneously detect both the neomycin-resistance gene and puromycin-resistance gene as an appropriate criterion for a positive result.…”

Avian Colibacillosis and Salmonellosis: A Closer Look at Epidemiology, Pathogenesis, Diagnosis, Control and Public Health Concerns

“…At slaughter, resistant strains from the gut readily soil poultry carcasses and as a result poultry meats are often contaminated with multiresistant E. coli [8][9][10][11][12][13][14]; likewise eggs become contaminated during laying [15]. Hence, resistant faecal E. coli from poultry can infect humans both directly and via food.…”