ABSTRACT
OBJECTIVES: Pseudomonas aeruginosa is an opportunistic pathogen with remarkable adaptation ability to thrive in diverse environmental conditions. This study aimed at phenotypic and molecular analysis of metallo beta lactamases (blaIMP, blaVIM, blaNDM-1 and blaSPM-1) and genetic diversity analysis among imipenem resistant clinical isolates of Pseudomonas aeruginosa. METHODS: This study was conducted from May 2017 to June 2018. The study included 187 Pseudomonas aeruginosa isolates collected from different clinical specimens from Peshawar, Pakistan. The isolates were analyzed for resistance to imipenem. Combined disc test (CDT) was then performed for phenotypic detection of metallo beta lactamases among imipenem resistant isolates of Pseudomonas aeruginosa. Molecular detection of metallo beta lactamases genes i.e. blaIMP, blaVIM, blaNDM-1 and blaSPM-1 was analyzed through polymerase chain reaction. Genetic diversity was determined through RAPD-PCR. RESULTS: MBL production was observed in 76% (n=19) isolates. The occurrence of MBL genes blaIMP, blaNDM-1 and blaVIM was 68% (n=17), 48% (n=12), and 4% (n=1) respectively. The blaSPM-1 gene was not detected. High genetic diversity was observed in current study. Out of 182 isolates 171 isolates showed different RAPD profiles (93.95% polymorphism); 160 were unique RAPD strains and based on similarity coefficient ≥ 80%, 22 isolates were clustered into 11 distinct clones. CONCLUSION: A high prevalence of blaIMP and blaNDM-1 among imipenem resistant isolates of Pseudomonas aeruginosa is alarming that calls for proper control and prevention strategies. RAPD technique was found to be a good genotyping technique when limited resources are available.
ABSTRACT
We report the prevalence of extended-spectrum ß-lactamases and carbapenemases in Escherichia coli isolated from retail chicken in Peshawar, Pakistan. One hundred E. coli isolates were recovered from retail chicken. Antibiotic susceptibility testing was carried out against ampicillin, chloramphenicol, kanamycin, nalidixic acid, cephalothin, gentamicin, sulfamethoxazole-trimethoprim, and streptomycin. Phenotypic detection of ß-lactamase production was analyzed through double disc synergy test using the antibiotics amoxicillin-clavulanate, cefotaxime, ceftazidime, cefepime, and aztreonam. Fifty multidrug-resistant isolates were screened for detection of sul1, aadA, cmlA, int, blaTEM, blaSHV, blaCTX-M, blaOXA-10, blaVIM, blaIMP, and blaNDM-1 genes. Resistance to ampicillin, nalidixic acid, kanamycin, streptomycin, cephalothin, sulfamethoxazole-trimethoprim, gentamicin, cefotaxime, ceftazidime, aztreonam, cefepime, amoxicillin-clavulanate, and chloramphenicol was 92, 91, 84, 73, 70, 67, 53, 48, 40, 39, 37, 36, and 23% respectively. Prevalence of sul1, aadA, cmlA, int, blaTEM, blaCTX-M, blaIMP, and blaNDM-1 was 78% ( n = 39), 76% ( n = 38), 20% ( n = 10), 90% ( n = 45), 74% ( n = 37), 94% ( n = 47), 22% ( n = 11), and 4% ( n = 2), respectively. blaSHV, blaOXA-10, and blaVIM were not detected. The coexistence of multiple antibiotic resistance genes in multidrug-resistant strains of E. coli is alarming. Hence, robust surveillance strategies should be developed with a focus on controlling the spread of antibiotic resistance genes via the food chain.
Subject(s)
Bacterial Proteins/metabolism , Chickens/microbiology , Drug Resistance, Multiple, Bacterial , Escherichia coli , beta-Lactamases/metabolism , Animals , Anti-Bacterial Agents/pharmacology , Escherichia coli/drug effects , Escherichia coli/enzymology , Food Microbiology , PakistanABSTRACT
Tuberculosis is a major infectious disease caused by Mycobacterium tuberculosis complex. Antimicrobial drugs are used to control TB infections. Molecular mechanisms controlling resistance to second-line drugs are not completely understood and no endogenous information is available regarding these mechanisms. The present study reports mutational analysis of rrs gene in Mycobacterium tuberculosis isolates collected from Khyber Pakhtunkhwa province of Pakistan. A total of 499 Mycobacterium tuberculosis isolates were analyzed for resistance against amikacin. Thirty resistant isolates were selected for mutational analysis in rrs gene. Among the 30 amikacin resistant isolates of Mycobacterium tuberculosis, 9 (30%) had mutation in the hotspot region of rrs gene. The predominant mutation was 1401A > G which was observed in 5 isolates. Maximum number of mutations was observed in isolate 6 and isolate 16 with six different mutations each. Mutations in isolate 6 included 1260G > A, 1278A > T, 1278_1279insT, 1300C > T, 1321G > A and 1445C > T. Mutation in isolate 16 included 1255_1256insA, 1364_1365insG, 1384_1385insA, 1880_1881insT, 1487G > A, and 1493delA. The mutation 1263G > A was observed in isolate 1. Isolate 2 had the 1484G > T mutation. The findings could be used as reference for future endures. It was evident from the results that mutations in rrs gene do not always contribute to amikacin resistance; hence, traditional drug susceptibility testing is still helpful for evaluation of such samples.
