Molecular detection of genes encoding resistance to tetracycline and determination of plasmid-mediated resistance to quinolones in avian pathogenic Escherichia coli in Sukabumi, Indonesia

Abstract

Aim:This study aimed to identify genes encoding resistance to tetracycline (TE) and plasmid-mediated resistance to quinolones in Escherichia coli isolates from clinical cases of avian colibacillosis in Sukabumi, Indonesia.Materials and Methods:A total of 25 E. coli archive isolates were collected in 2013-2017 from clinical cases of avian colibacillosis in Sukabumi, Indonesia. All isolates were tested for TE and quinolone resistance using the disk diffusion method. TE -resistant E. coli isolates were screened for the presence of tet(A) and tet(B) genes by single polymerase chain reaction (PCR). The qnr(A), qnr(B), and qnr(S) genes were detected by multiplex PCR in quinolone-resistant E. coli isolates.Results:Result of this study shows that 19 of 25 (76%) E. coli isolates are resistant to oxytetracycline and 64% are resistant to TE; among them, 63.2% and 31.5% were positive tet(A) and tet(B), respectively. 13 out of 25 (52%) are resistant to ciprofloxacin and 36% are resistant to enrofloxacin either norfloxacin; among them, 61.6% were positive qnr(A), 7.7% were positive qnr(B), 23% were positive qnr(S), and 7.7% were positive both of qnr(A) and qnr(S).Conclusion:This study shows that a few pathogens of E. coli are resistant to TE and quinolone. The frequency of tet and qnr genes that are responsible for this resistance among avian pathogenic E. coli isolates in Sukabumi, Indonesia, was high.

Research Note: Characterization of antibiotic resistant phenotypes and linked genes of Escherichia coli and Klebsiella pneumoniae from healthy broiler chickens, Karnataka, India

“…Pure bacterial colonies grown on blood agar (HiMedia, India) at 37°C for 24 hr were used for DNA extraction by QIAamp DNA mini kit (QIAGEN, Dusseldorf, Germany). ESBLs - bla TEM , bla SHV and bla CTX-M ( Sinha et al., 2015 ), bla CTX-M-15 ( Zowawi et al., 2014 ); plasmid-mediated quinolone resistance - qnrA, qnrB, qnrS ( Huang et al., 2009 ) and tetracycline efflux - tet(A), tet(B) ( Kurnia et al., 2018 ) genes were analyzed by PCR. The PCR reaction was as follows – 25 μL volume contained 12.5 μL KAPA Taq Hotstart 2X master mix (Kapa Biosystems, Wilmington, MA, USA), 1 μL each primer, 2μL DNA template, and 8.5 μL nuclease-free water.…”

Avian Pathogenic Escherichia coli (APEC): An Overview of Virulence and Pathogenesis Factors, Zoonotic Potential, and Control Strategies

“…Many antibiotics belonging to different classes, such as tetracyclines (tetracycline, oxytetracycline, chlortetracycline), sulfonamides (sulfadimethoxine, trimethoprim, sulfadiazine, sulfamethazine, sulfaquinoxaline, ormethoprim), aminoglycosides (apramycin, gentamicin, neomycin, spectinomycin), penicillins (amoxicillin, ampicillin), cephalosporins (ceftiofur), quinolones (danofloxacin, sarafloxacin, enrofloxacin), polymyxins (colistin), chloramphenicols (florfenicol), macrolides (erythromycin), and lincosamides (lincomycin) have been used in poultry industry worldwide for the control of APEC infections [ 23 ]. However, APEC resistance to multiple antibiotics has been reported [ 32 , 34 , 35 , 51 , 53 , 54 , 70 , 71 , 72 , 73 , 107 , 108 , 108 , 146 , 147 , 148 , 149 , 150 , 151 , 152 , 153 , 154 , 155 , 156 , 157 , 158 , 159 , 160 , 161 , 162 , 163 , 164 , 165 , 166 , 167 , 168 , 169 , 170 , 171 , 172 ], which limits the use of these antibiotics and suggests a challenge ahead in using these antibiotics. Table 2 provides a summary of antibiotic resistance and resistance genes (mechanisms) reported worldwid...…”

Isolation and molecular characterization of fowl aviadenovirus associated with inclusion body hepatitis from poultry in Banten and West Java, Indonesia

“…Amplified samples were analyzed by electrophoresis in 1.5% agarose gel and stained by ethidium bromide. A molecular weight marker with 100 bp (Geneaid, Taiwan) was used as a standard size [ 10 , 11 ].…”