scholarly journals Evaluation on the Use of β-Lactamase and Aminoglycoside Modifying Enzyme Gene Sequences as Markers for the Early Detection of Antibiotic Resistance Profile ofPseudomonas aeruginosa

2004 ◽  
Vol 20 (6) ◽  
pp. 317-323 ◽  
Author(s):  
Victor A. Doss ◽  
S. Parvathi ◽  
B. Appala Raju ◽  
N. Abitha Devi

Pseudomonas aeruginosais one of the major causes of infections including the hospital acquired (Nosocomial) infections. Detection of them and their antibiotic resistance profile by conventional method takes about three days. Recently, DNA based diagnostic methods are being used for the identification of the pathogens. Hence we have tested a rapid and sensitive method using DNA sequences as markers for detecting the presence of three genes coding for the enzymes that inactivate the two most commonly used Anti-pseudomonadal drugs such as β-lactam antibiotics (Penicillin, and its derivatives) and Aminoglycosides such as Gentamicin, Tobramycin, Amikacin, Streptomycin. The internal region of these genes were used for designing and synthesizing primers and these primers were used in Polymerase Chain Reaction (PCR) to screen for the presence of these genes in the clinical isolates and to label them non-radioactively with Biotin. They in turn were used to detect the presence of the antibiotic resistance genes in the clinical isolates by hybridization. The specificity (ratio of positive results obtained in both methods and the sensitivity (the minimum amount of sample DNA and the labeled probe required for the tests) were evaluated.

2021 ◽  
Vol 14 (4) ◽  
pp. 1847-1854
Author(s):  
Vaibhavi Patel

A simple explanation for antimicrobial-resistant opportunistic infections in immunocompromised patients is Klebsiella pneumoniae which gradually being associated in insidious infections globally with high mortality rate. Eight hundred fifty-six antibiotic resistant K. pneumoniae isolates were collected over 3 years period (from different wards and different specimens) from the Microbiology department of C.U. Shah hospital, whose AST checked by Kirby Bauer disk diffusion method. To study AMR genes, virulome, interference of virulence gene with resistance gene, phylogenomic; 6 clinical isolates were proceeded for whole genome sequencing and bio informatics analysis. Klebsiella pneumoniae is a multidrug-resistant (MDR) opportunistic and one of delegate of ESKAPE pathogens groups. This pathogen causes nosocomial infections, urinary tract infections, liver abscesses, wound infections, meningitis. These strains obtain a multidrug resistant phenotype by way of horizontal transfer of ARG transported by either transposons or plasmids. This transfer is generally facilitated by Integrons. In this study antibiotic resistance profile and antibiotic resistance genes analysis as well as virulence gene of K. pneumoniae strains were investigated. The study was carried out using 853 clinical isolates collected during 3 years from C.U. Shah hospital of Surendranagar. Antibiotic resistance profile test was carried out by the VITEK 2 against 21 antibiotics. Out of that 6 samples were proceed for DNA extraction, WGS illumina sequencer and analysis of those raw sequences by TORMES pipeline. In this study antibiotic resistance profile included 13 beta lactam antibiotics which classified under 3 class (Penicillin, Cephalosporin, Carbapenem) of beta lactam and in AMR gene study got total 15 different ESBL resistance genes from 6 different klebsiella pneumoniae strain. All these genes detected with more than 90% identity by CARD. (TORMES Pipeline) CTX-M-15, NDM-5, OKP-B-6, PDC-2, OXA-1, OXA-181, OXA-362, OXA-50, OXA-9, SHV-1, SHV-11, SHV-187, TEM-1, TEM-150. In this study, we’ve analyzed the pattern of antibiotic resistance pattern as a phenotypic characteristic and antibiotic resistance genes as genotypic characteristic and co related the results. As multidrug resistance is a worrying matter, constant observation and regular clinical recognition of resistant bacteria are essential to avoid terrible public health incidents. So, our data should be inferred as a warning for need for prevention and control of the MDR K. pneumoniae in hospital settings.


Antibiotics ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 575
Author(s):  
Emi Nishimura ◽  
Masateru Nishiyama ◽  
Kei Nukazawa ◽  
Yoshihiro Suzuki

Information on the actual existence of antibiotic-resistant bacteria in rivers where sewage, urban wastewater, and livestock wastewater do not load is essential to prevent the spread of antibiotic-resistant bacteria in water environments. This study compared the antibiotic resistance profile of Escherichia coli upstream and downstream of human habitation. The survey was conducted in the summer, winter, and spring seasons. Resistance to one or more antibiotics at upstream and downstream sites was on average 18% and 20%, respectively, and no significant difference was observed between the survey sites. The resistance rates at the upstream site (total of 98 isolated strains) to each antibiotic were cefazolin 17%, tetracycline 12%, and ampicillin 8%, in descending order. Conversely, for the downstream site (total of 89 isolated strains), the rates were ampicillin 16%, cefazolin 16%, and tetracycline 1% in descending order. The resistance rate of tetracycline in the downstream site was significantly lower than that of the upstream site. Furthermore, phylogenetic analysis revealed that many strains showed different resistance profiles even in the same cluster of the Pulsed-Field Gel Electrophoresis (PFGE) pattern. Moreover, the resistance profiles differed in the same cluster of the upstream and the downstream sites. In flowing from the upstream to the downstream site, it is plausible that E. coli transmitted or lacked the antibiotic resistance gene.


Author(s):  
Anupama Bhardwaj ◽  
Jagtar Singh ◽  
Sonia Chaman ◽  
Amit Joshi

Objective: The objective of this study is to make sure biotreatment process used for treatment of dairy wastewater (DWW) is safe for human and its surrounding environment; microbes were evaluated for their antibiotic resistance profile against commonly prescribed antibiotics. Methods: Microbes were isolated using spread plating and streaking method and used to treat DWW. Reduction in organic load in DWW was determined by comparing physicochemical parameters (PCP) of DWW before and after treatment process. After selection of efficient microbial isolates, they were evaluated for their antibiotic resistance profile using antibiotic disc diffusion method. Results: In this work, 53 microbes were isolated from DWW, and these microbial isolates were screened for DWW degradation capacity by analyzing PCP. Four microbial isolates E3, E5, E11 (bacterial isolates) and F5 (fungal isolate) showed highest reduction in chemical oxygen demand (COD), biological oxygen demand (BOD), and dissolved oxygen (DO) were selected for profound degradation of DWW under optimized conditions. Efficient four microbial isolates individually performed better under anaerobic conditions by showing maximum reduction 84%, 75%, and 77% in COD, BOD, and DO, respectively. After 72 hrs of antibiotic susceptibility testing, E3 strain had shown 100%, E5 90%, E11 70%, and F5 80% susceptibility to antibiotics. Conclusion: The present study concluded that four microbial isolates had the potential of reducing the organic load of DWW along with lessor or negligible adverse effect on human or its surrounding environment and they appear to be most promising strains for treatment of DWW. 


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