BackgroundThe increasing and rapid spread of metallo-beta-lactamase (MBL) producing Enterobacteriaceae, particularly Escherichia coli and Klebsiella pneumoniae represents an emerging public health threat. However, limited data is available on MBL production in clinical isolates of E. coli and K. pneumoniae from Nepal. We have therefore undertaken this study to ascertain the incidence of MBL production in clinical isolates of E. coli and K. pneumoniae at a tertiary care teaching hospital in central Nepal.MethodsA total of 401 consecutive, non-duplicate isolates of E. coli (n = 216) and K. pneumoniae (n = 185) were recovered from various clinical samples between July and December, 2012. These isolates were screened for the detection of carbapenemase production on the basis of their reduced susceptibility to meropenem or ertapenem by the disc diffusion method. The screened isolates were further phenotypically studied for carbapenemase production by modified Hodge test (MHT). MBL production was detected by performing combined disc test by using imipenem discs with and without ethylenediaminetetraacetic acid (EDTA), which chelates zinc required for MBL activity.ResultsOut of 216 E. coli isolates, a total of 41 isolates (18.98%) and out of 185 K. pneumoniae isolates, a total of 39 isolates (21.08%) were suspected to be carbapenemase- producers on the basis of their reduced susceptibility to meropenem or ertapenem. Interestingly, all the initially suspected isolates of E. coli and K. pneumoniae for carbapenemase production were found to be positive in both MHT and combined disc test. However, few weakly positive reactions were observed in MHT. All the MBL producing isolates were multidrug-resistant (MDR). In addition, 75.60% E. coli and 71.79% of K. pneumoniae isolates producing MBL were found to be “pandrug- resistant”.ConclusionsOur findings showed MBL production in a considerable number of E. coli and K. pneumoniae isolates with MDR and pandrug-resistant phenotypes. Combined disc method can provide a sensible choice for phenotypic detection of MBL production in clinical microbiology laboratories as detection of MBL in bacterial isolates is indispensable for establishing the effective antibiotic policies and infection control strategies in the hospital setting.

Background: Pseudomonas aeruginosa is an opportunistic pathogen which causes most of the chronic infection in humans. This study was undertaken to determine the prevalence rate of Pseudomonas aeruginosa that is isolated from various clinical specimens along with its antibiotic susceptibility pattern.Methods: This descriptive cross sectional study was conducted in Kathmandu Medical College and Teaching Hospital (KMCTH) from February to May 2018. Pseudomonas aeruginosa isolated from various clinical specimens were processed in clinical laboratory, Department of Microbiology, KMCTH. Isolation, identification and sensitivity of Pseudomonas aeruginosa to antibiotics were measured.Results: A total of 7527 samples were been processed of which 46 isolates of Pseudomonas aeruginosa were obtained. Pseudomonas aeruginosa was isolated mainly from Pus, Wound swab, Sputum and Tracheal aspirate. Here 63.04% Pseudomonas aeruginosa isolates were resistant to Ceftazidime, 65.21% to Cefixime, 56.52% to Ceftriaxone and Cefotaxime followed by 56.52% to Piperacillin. Furthermore, the current study reveals antibiotics like Imipenem, Meropenem, Piperacillin/Tazobactam, Ciprofloxacin, Gentamicin, Amikacin and Tobramycin were found to be good choice for the treatment of infection caused by this organism.Conclusions: Continuous monitoring of antibiotic susceptibility pattern of Pseudomonas aeruginosa is essential and rational treatment regimens prescription by the clinicians is required to limit the spread of antimicrobial resistance.Keywords: Antibiotic resistance; clinical isolates; Pseudomonas aeruginosa.

Coronavirus disease-19 is caused by severe acute respiratory syndrome coronavirus-2 (SARSCoV-2). Specimen quality, and proper transportation is vital for accurate diagnosis. This standard operating procedure is designed to educate the clinicians, nurses, paramedics, and laboratory personnel regarding proper methods of sample collection, packaging, and transportation. Nasopharyngeal swabs and/or oropharyngeal swabs should be collected for real-time quantitative polymerase chain reaction to detect SARS-CoV-2. The sample should be collected wearing proper personal protective equipment, packed in a triple packaging system, and transported maintaining cold chain.

Biofilm refers to the complex, sessile communities of microbes found either attached to a surface or buried firmly in an extracellular matrix as aggregates. Microbial flora which produces biofilm manifests an altered growth rate and transcribes genes that provide them resistance to antimicrobial and host immune systems. Biofilms protect the invading bacteria against the immune system of the host via impaired activation of phagocytes and the complement system. Biofilm-producing isolates showed greater multidrug resistance than non-biofilm producers. Biofilm causes antibiotic resistance through processes like chromosomally encoded resistant genes, restriction of antibiotics, reduction of growth rate, and host immunity. Biofilm formation is responsible for the development of superbugs like methicillin-resistant Staphylococcus aureus, vancomycin-resistant Staphylococcus aureus, and metallo-beta-lactamase producing Pseudomonas aeruginosa. Regular monitoring of antimicrobial resistance and maintaining hygiene, especially in hospitalized patients are required to control biofilm-related infections in order to prevent antimicrobial resistance.

Introduction: Gram negative bacilli are the important causes of common clinical infections. Carbapenem resistant Enterobacteriaceae are considered as important public health threat and is classified as urgent by the Centers of Disease Control and Prevention because of their progressive geographic dissemination and limited therapeutic alternatives. This study was done to find out the resistance pattern of Carbapenem among Enterobacteriaceae.Methods: The descriptive cross-sectional study was carried out in Clinical Microbiology laboratory from February 2018 to May 2018 after ethical approval. Organism was identified on the basis of its microscopic observation by performing Gram’s stain and by identification of morphology after its growth in culture media followed by its biochemical reactions. Antibiotic sensitivity test of isolated pathogens was done using Muller Hinton Agar by the standard disk diffusion technique of Kirby-Bauer method.Results: In our study, total 1055 sample belongs to the family Enterobacteriaceae. From the family Enterobactericeae, 348 (27%) of the bacilli were found to be Carbapenem resistant. Among which most common bacteria was Klebsiella pneumoniae followed by Escherichia coli. All strains of Carbapenem resistant Enterobacteriaceae were sensitive to Colistin, Polymyxin B and Tigecycline. Conclusions: Among Enterobacteriaceae, around one-third of the bacterial isolates were Carbapenem resistant. However, to reduce drug resistance antimicrobial stewardship programme and proper infection control measures is required.