Publications by authors named "Shyam Kumar Mishra"

24 Publications

  • Page 1 of 1

Comparative study of Amsel's criteria and Nugent scoring for diagnosis of bacterial vaginosis in a tertiary care hospital, Nepal.

BMC Infect Dis 2021 Aug 17;21(1):825. Epub 2021 Aug 17.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: The most common pathological cause of abnormal vaginal discharge in reproductive-aged women is bacterial vaginosis (BV). Amsel's criteria and Nugent scoring systems are commonly employed approaches for the diagnosis of BV. Despite the Nugent scoring system being the gold standard method for diagnosing BV, Amsel's criteria are generally preferred in clinical setup owing to the fact Nugent scoring requires considerable time and expert microscopist. This study was conducted to determine the diagnostic value of Amsel's criteria by comparing it with the Nugent scoring system.

Methods: This was a descriptive cross-sectional study conducted at Tribhuvan University Teaching Hospital, Nepal from October 2016 to September 2017. Vaginal specimens were collected from a total of 141 women presenting with abnormal vaginal discharge. The sensitivity, specificity, positive predictive value, and negative predictive value of Amsel's criteria were calculated, and each component of Amsel's criteria was compared to the Nugent scoring system.

Results: The sensitivity, specificity, positive predictive value, and negative predictive value of Amsel's criteria were 50%, 98.2%, 87.5%, and 88.8% respectively. The clue cells showed 100% specificity and vaginal discharge with pH > 4.5 had 89.3% sensitivity while compared with Nugent's scoring system.

Conclusions: Amsel's criteria can be used as an adjunct method to Nugent scoring for the diagnosis of BV in the hands of skilled manpower in resources limited countries. The presence of clue cell and positive whiff test of Amsel's criteria shows good match with Nugent's score.
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http://dx.doi.org/10.1186/s12879-021-06562-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8369704PMC
August 2021

Characterisation of ESKAPE Pathogens with Special Reference to Multidrug Resistance and Biofilm Production in a Nepalese Hospital.

Infect Drug Resist 2021 14;14:2201-2212. Epub 2021 Jun 14.

St. Xavier's College (Tribhuvan University), Department of Microbiology, Kathmandu, Nepal.

Background: "ESKAPE" is an acronym for a group of life-threatening nosocomial pathogens, viz, and spp. Global efforts on controlling multidrug-resistant (MDR) organisms have been hampered by their ability to escape antibacterial drugs. This study was undertaken to determine the prevalence of ESKAPE pathogens with prime focus on biofilm production and antibiotic resistance.

Methods: A total of 8756 clinical samples were processed for the isolation and identification of ESKAPE pathogens following standard microbiological procedures. These isolates were subjected to antimicrobial sensitivity test as per Clinical and Laboratory Standards Institute (CLSI) guidelines. Test for MDR, extended-spectrum β-lactamase (ESBL), metallo-β-lactamase (MBL), methicillin-resistant (MRSA), vancomycin-resistant (VRE) was done by the disk diffusion and E-test methods. In the case of VRE molecular detection was done for and genes. All the isolates were processed for biofilm detection by the tube adherence method.

Results: The percentage distribution of was 5.5%, 33.4%, 33.0%, 8.6%, 18.6%, and 0.9%. MRSA was 57.6%, and vancomycin resistance among was 20%. ESBL- and MBL-producing were 16.1%, and 8.1%, 10.3% each and 10.7% and 8.3%, respectively. A total of 42.3% of isolates were biofilm producers. Linezolid was the drug of choice for VRE. Ampicillin-sulbactam was most useful against apart from polymyxins, whereas piperacillin-tazobactam was effective against other Gram-negative bacteria. gene was detected in all the VRE isolates.

Conclusion: This study estimates the burden of the ESKAPE organisms and their antimicrobial resistance pattern in a hospital setting. A high percentage of drug resistance and biofilm production was noted; hence antimicrobial resistance surveillance targeting ESKAPE pathogens should be incorporated in the infection control policy in Nepal.
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http://dx.doi.org/10.2147/IDR.S306688DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8214009PMC
June 2021

Biofilm Formation by Pathogens Causing Ventilator-Associated Pneumonia at Intensive Care Units in a Tertiary Care Hospital: An Armor for Refuge.

Biomed Res Int 2021 28;2021:8817700. Epub 2021 May 28.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Emerging threat of drug resistance among pathogens causing ventilator-associated pneumonia (VAP) has resulted in higher hospital costs, longer hospital stays, and increased hospital mortality. Biofilms in the endotracheal tube of ventilated patients act as protective shield from host immunity. They induce chronic and recurrent infections that defy common antibiotics. This study intended to determine the biofilm produced by pathogens causing VAP and their relation with drug resistance.

Methods: Bronchoalveolar lavage and deep tracheal aspirates ( = 70) were obtained from the patients mechanically ventilated for more than 48 hours in the intensive care units of Tribhuvan University Teaching Hospital, Kathmandu, and processed according to the protocol of the American Society for Microbiology (ASM). Antibiotic susceptibility testing was done following Clinical and Laboratory Standards Institute (CLSI) 2017 guidelines. Biofilm formation was determined using the microtiter plate method described by Christensen and modified by Stepanovoic et al.

Results: Significant microbial growth was seen in 78.6% of the total samples with 52.7% monomicrobial, 45.5% polymicrobial, and 1.8% fungal infection. Among the 71 isolates obtained, bulk was gram-negative ( = 64, 90.1%). (31.0%) was the predominant isolate followed by complex (16.9%), (16.9%), (15.5%), (7.0%), (5.6%), (2.8%), (1.4%), complex (1.4%), and (1.4%). Of the total isolates, 56.3% were biofilm producers. Multidrug-resistant (MDR) organisms, extended-spectrum -lactamase (ESBL), and metallo--lactamase (MBL) producers were preeminent among the biofilm producers. The highest producer of biofilm was (19.7%). Among gram-negative biofilm producers, 42.2% were MDR, 21.9% were ESBL producers, and 7.8% were MBL producers.

Conclusion: Gram-negative nonfermenter bacteria account for the bulk of nosocomial pneumonia. MDR, ESBL, and MBL production was preponderant among the biofilm producers. The rampant spread of drug resistance among biofilm producers is summoning novel interventions to combat multidrug resistance.
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http://dx.doi.org/10.1155/2021/8817700DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8179767PMC
May 2021

In vitro Antimicrobial Synergy Testing of Extensively Drug-Resistant Clinical Isolates at an Organ Transplant Center in Nepal.

Infect Drug Resist 2021 30;14:1669-1677. Epub 2021 Apr 30.

School of Optometry and Vision Science, Faculty of Medicine and Health, UNSW, Sydney, NSW, Australia.

Purpose: Inappropriate use of broad-spectrum antibiotics contributes to the emergence of multidrug-resistant (MDR) bacteria. Finding novel antimicrobial agents and strategies based on synergistic combinations are essential to combat MDR infections. This study was designed to determine in vitro synergy of different antimicrobials against extensively drug-resistant (XDR) Gram-negative clinical isolates.

Methods: A descriptive, cross-sectional study was conducted at Human Organ Transplant Center, Nepal, for five months. Clinical isolates were checked for their drug-resistance properties including extended-spectrum beta-lactamase- (ESBL-) and metallo-beta-lactamase- (MBL-) production. The XDR isolates were further tested for antimicrobial synergy, and the results were interpreted as synergistic, additive, indifferent or antagonistic determining fractional inhibitory concentration of the antibiotics.

Results: Out of total 1155 clinical samples, 308 showed significant growth. was the most common isolate (n=142) followed by complex, and miscellaneous bacteria. Out of the culture positive isolates, 21.4% were MDR and 10.06% were XDR. The XDR population comprised (18.42%), (9.86%), complex (7.41%) and (4.17%). Among the culture positive isolates, 4.5% and 5.8% were ESBL- and MBL-producers, respectively. Colistin, polymyxin B, and tigecycline were the antibiotics effective in majority of MDR isolates as compared to carbapenems. The combination of antibiotics - meropenem and colistin showed the highest proportion of "synergy" among all XDR whereas the combination of amikacin and colistin showed synergistic effect in XDR .

Conclusion: A significant proportion of isolates were MDR among which a large fraction was XDR. The combination of meropenem, amikacin and colistin with one another in pair showed beneficial activity in vitro. Such combinations can be utilized as effective therapy for XDR infections. Further studies are required to confirm these findings, and accordingly treatment protocols should be developed in the management of such infections.
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http://dx.doi.org/10.2147/IDR.S309531DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8096450PMC
April 2021

Carbapenem Resistance in Non-Fermentative Gram-Negative Bacilli Isolated from Intensive Care Unit Patients of a Referral Hospital.

J Nepal Health Res Counc 2021 Apr 23;19(1):55-61. Epub 2021 Apr 23.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Non-fermentative Gram-negative bacilli or non-fermenters are opportunistic pathogens associated with serious infections in intensive care unit patients. Although carbapenems were considered as a backbone of treatment for life-threatening infections, these bacteria are increasingly acquiring resistance to carbapenems. Carbapenem-resistant Acinetobacter baumannii and Pseudomonas aeruginosa are prioritized as critical pathogens by the World Health Organization. The objective of the study was to document the status of carbapenem-resistant and carbapenemase-producing non-fermenters isolated from intensive care unit patients.

Methods: This study was conducted at Tribhuvan University Teaching Hospital, Kathmandu, Nepal. The clinical specimens collected from intensive care unit patients were processed for isolation and identification of non-fermenters and antibiotic susceptibility profile of bacterial isolates was determined. The multidrug-resistant isolates were identified and carbapenemase enzyme was detected in the carbapenem-resistant isolates.

Results: A total of 157 non-fermenters were isolated from 1063 samples which included Acinetobacter species (n=85), Pseudomonas aeruginosa (n=55), Burkholderia cepacia complex (n=15), and Stenotrophomonas maltophilia (n=2). Carbapenem resistance was reported in 85.9%, 72.7%, and 33.3% of Acinetobacter species, Pseudomonas aeruginosa, and Burkholderia cepacia complex, respectively. Among total non-fermenters, 91.1% isolates were multidrug-resistant and 60.8% carbapenem-resistant isolates were carbapenemase producers. The carbapenem-resistant isolates demonstrated an extremely high degree of resistance than carbapenem-susceptible isolates towards other antimicrobial classes.

Conclusions: This study reported high rates of carbapenem-resistant, carbapenemase-producing, and multidrug-resistant non-fermenters isolates. Therefore, preventing the spread of these superbugs among the critically ill patients in intensive care units should be a major initiative in hospitals.
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http://dx.doi.org/10.33314/jnhrc.v19i1.3240DOI Listing
April 2021

Ventilator-associated pneumonia among ICU patients in WHO Southeast Asian region: A systematic review.

PLoS One 2021 9;16(3):e0247832. Epub 2021 Mar 9.

Department of Microbiology, Institute of Medicine, Tribhuvan University, Kathmandu, Nepal.

Ventilator-associated pneumonia (VAP) is one of the most frequent ICU-acquired infections and a leading cause of death among patients in Intensive Care Unit (ICU). The South East Asian Region is a part of the world with limited health resources where infectious diseases are still underestimated. We aimed to review the literature in this part of the world to describe incidence, mortality and microbiological evidence of VAP and explore preventive and control strategies. We selected 24 peer-reviewed articles published from January 1, 2000 to September 1, 2020 from electronic databases and manual searching for observational studies among adult patients diagnosed with VAP expressed per thousand days admitted in ICU. The VAP rates ranged from 2.13 to 116 per thousand days, varying among different countries of this region. A significant rate of mortality was observed in 13 studies ranging from 16.2% to 74.1%. Gram negative organisms like Acinetobacter spp., Pseudomonas aeruginosa and Klebsiella pneumoniae and Gram-positive organisms like Staphylococcus aureus and Enterococcus species were frequently found. Our findings suggest an alarming situation of VAP among patients of most of the countries of this region with increasing incidence, mortality and antibiotic resistance. Thus, there is an urgent need for cost effective control and preventive measures like interventional studies and educational programs on staff training, hand hygiene, awareness on antibiotic resistance, implementation of antibiotic stewardship programs and appropriate use of ventilator bundle approach.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0247832PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7942996PMC
March 2021

How Well the Government of Nepal Is Responding to COVID-19? An Experience From a Resource-Limited Country to Confront Unprecedented Pandemic.

Front Public Health 2021;9:597808. Epub 2021 Feb 17.

Centre for Primary Health Care and Equity, University of New South Wales, Sydney, NSW, Australia.

COVID-19, caused by SARS-CoV-2, was first reported in Wuhan, China and is now a pandemic affecting over 218 countries and territories around the world. Nepal has been severely affected by it, with an increasing number of confirmed cases and casualties in recent days, even after 8 months of the first case detected in China. As of 26 November 2020, there were over 227,600 confirmed cases of COVID in Nepal with 209,435 recovered cases and 1,412 deaths. This study aimed to compile public data available from the Ministry of Health and Population (MoHP), Government of Nepal (GoN) and analyse the data of 104 deceased COVID-19 patients using IBM SPSS (Version 25.0). Additionally, this study also aimed to provide critical insights on response of the GoN to COVID-19 and way forward to confront unprecedented pandemic. Figures and maps were created using the Origin Lab (Version 2018) and QGIS (Version 3.10.8). Most of the reported cases were from Bagmati Province, the location of Nepal's capital city, Kathmandu. Among deceased cases, >69% of the patients were male and patients ≥54 years accounted for 67.9% ( = 923). Preliminary findings showed respiratory illness, diabetes, and chronic kidney diseases were the most common comorbid conditions associated with COVID-19 deaths in Nepal. Despite some efforts in the 8 months since the first case was detected, the government's response so far has been insufficient. Since the government eased the lockdown in July 2020, Nepal is facing a flood of COVID-19 cases. If no aggressive actions are taken, the epidemic is likely to result in significant morbidity and mortality in Nepal. The best way to curb the effect of the ongoing pandemic in a resource-limited country like Nepal is to increase testing, tracing, and isolation capacity, and to set up quality quarantine centers throughout the nation. A comprehensive health literacy campaign, quality care of older adults and those with comorbidity will also result in the effective management of the ongoing pandemic.
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http://dx.doi.org/10.3389/fpubh.2021.597808DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7925847PMC
March 2021

Prevalence of vancomycin-resistant enterococci in Asia-A systematic review and meta-analysis.

J Clin Pharm Ther 2021 Oct 25;46(5):1226-1237. Epub 2021 Feb 25.

Department of Microbiology, Institute of Medicine, Tribhuvan University, Kathmandu, Nepal.

What Is Known And Objective: The rise of vancomycin-resistant enterococci (VRE) has been a major health problem in most countries of the world including Asia, since its discovery. There is a paucity of data on VRE in many countries of Asia as well as limited pooled estimates. Therefore, we performed a systematic review and meta-analysis to estimate a pooled prevalence of VRE in Asia.

Methods: A literature search in electronic databases like PubMed, Embase and Google Scholar and manual searching of references and grey literature, comprising the information on the prevalence of VRE with at least two species of enterococci, conducted in different countries of Asia from January 1, 2000, to September 20, 2020, was done. The random-effect model and 95% CIs was used to calculate the pooled prevalence. Subgroup, sensitivity and meta-regression analyses were performed to address heterogeneity while Egger's test for publication bias.

Results And Discussions: We identified 39 studies, comprising a total of 11,875 enterococcal isolates. The result of the analysis showed that the pooled prevalence of VRE in Asia was 8.10% (95% CI; 7-9; I  = 93.79%; p < 0.001). Resistance to vancomycin was greater among strains of E. faecium compared to the strains of E. faecalis (22.40% vs. 3.70%). Amongst various regions of Asia, the highest prevalence of VRE was found in the Western Asian region and the lowest in the South-east Asian region. Moreover, the rate of VRE was higher than most European countries and lower than USA.

What Is New And Conclusions: With an upsurge of VRE in Asia in recent years, efficient infection control programmes, robust surveillance systems and adherence to antibiotic stewardship are paramount to halt the further rise of VRE.
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http://dx.doi.org/10.1111/jcpt.13383DOI Listing
October 2021

Bacteria on Medical Professionals' White Coats in a University Hospital.

Can J Infect Dis Med Microbiol 2020 29;2020:5957284. Epub 2020 Oct 29.

Department of Microbiology, Institute of Medicine, Tribhuvan University, Kathmandu, Nepal.

The transient contamination of medical professional's attires including white coats is one of the major vehicles for the horizontal transmission of microorganisms in the hospital environment. This study was carried out to determine the degree of contamination by bacterial agents on the white coats in a tertiary care hospital in Nepal. Sterilized uniforms with fabric patches of 10 cm × 15 cm size attached to the right and left pockets were distributed to 12 nurses of six different wards of a teaching hospital at the beginning of their work shift. Worn coats were collected at the end of the shifts and the patches were subjected for total bacterial count and identification of selected bacterial pathogens, as prioritized by the World Health Organization (WHO). Fifty percent of the sampled swatches were found to be contaminated by pathogenic bacteria. The average colony growth per square inch of the patch was 524 and 857 during first and second workdays, respectively, indicating an increase of 63.6% in colony counts. The pathogens detected on patches were , , , and sp. Additional bacteria identified included sp., sp., and coagulase-negative (CoNS). The nurses working in the maternity department had their white coats highly contaminated with bacteria. On the other hand, the least bacterial contamination was recorded from the nurses of the surgery ward. One isolate from the maternity ward was resistant to methicillin. This study showed that pathogens belonging to the WHO list of critical priority and high priority have been isolated from white coats of nurses, thus posing the risk of transmission to patients. White coats must be worn, maintained, and washed properly to reduce bacterial contamination load and to prevent cross-contamination of potential superbugs. The practice of wearing white coats outside the healthcare zone should be strictly discouraged.
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http://dx.doi.org/10.1155/2020/5957284DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7644330PMC
October 2020

A Syndemic Perspective on the Management of Non-communicable Diseases Amid the COVID-19 Pandemic in Low- and Middle-Income Countries.

Front Public Health 2020 25;8:508. Epub 2020 Sep 25.

School of Optometry and Vision Science, Faculty of Science, University of New South Wales, Sydney, NSW, Australia.

The global coronavirus disease (COVID-19) pandemic has greatly affected the lives of people living with non-communicable diseases (PLWNCDs). The health of PLWNCDs worsens when synergistic epidemics or "syndemics" occur due to the interaction between socioecological and biological factors, resulting in adverse outcomes. These interactions can affect the physical, emotional, and social well-being of PLWNCDs. In this paper, we discuss the effects of the COVID-19 syndemic on PLWNCDs, particularly how it has exposed them to NCD risk factors and disrupted essential public health services. We conclude by reflecting on strategies and policies that deal with the COVID-19 syndemic among PLWNCDs in low- and middle-income countries.
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http://dx.doi.org/10.3389/fpubh.2020.00508DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7545493PMC
May 2021

Emergence of multidrug-resistant non-fermentative gram negative bacterial infection in hospitalized patients in a tertiary care center of Nepal.

BMC Res Notes 2020 Jul 2;13(1):319. Epub 2020 Jul 2.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Objective: This study was designed for the characterization and establishment of antibiotic susceptibility profiles of non-fermentative gram negative bacteria isolated from hospitalized patients in a tertiary care hospital of Nepal.

Results: A total of 402 non-fermentative gram negative bacteria was isolated in 1486 culture-positive cases from 6216 different clinical samples obtained from hospitalized patients. Among total non-fermentative gram negative bacterial isolates, the highest number was recovered from specimens collected from lower respiratory tract infections (n = 173, 43.0%) of hospitalized patients followed by pus/swab samples (n = 99, 24.6%) and urinary tract infections (n = 49, 12.2%). The most common non-fermentative gram negative bacteria identified were Acinetobacter baumannii (n = 177, 44.0%), Pseudomonas aeruginosa (n = 161, 40.1%) and Burkholderia cepacia complex (n = 33, 8.2%). These bacterial isolates exhibited a higher rate of insusceptibility to beta-lactam antibiotics, fluoroquinolones, and aminoglycosides. On the other hand, all the isolates of P. aeruginosa and A. baumannii were completely susceptible to colistin sulfate and polymyxin B. Among total isolates, 78.1% (n = 314) were multidrug-resistant with a high rate of multidrug-resistant among A. baumannii (91.0%).
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http://dx.doi.org/10.1186/s13104-020-05163-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7330544PMC
July 2020

Burden of Multidrug-Resistant Infection in Hospitalized Patients in a Tertiary Care Hospital of Nepal.

Infect Drug Resist 2020 3;13:725-732. Epub 2020 Mar 3.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Introduction: has emerged as a significant multidrug-resistant (MDR) nosocomial pathogen worldwide and is responsible for various healthcare-associated infections. The MDR strains have been reported increasingly during the last decades in hospitalized patients. They have developed resistance to most of the available antibiotics and are known to produce various acquired β-lactamases. The β-lactamase producing strains have a potential for rapid dissemination in hospital settings, as it is often plasmid-mediated. The Infectious Diseases Society of America (ISDA) stated as one of the "red alert" pathogens that greatly threatens the utility of our current antibacterial armamentarium. The study attempted to investigate the spectrum and antimicrobial resistance among MDR and their potential implications in hospitalized patients in a tertiary care hospital of Nepal.

Methods: This study was conducted at Tribhuvan University Teaching Hospital (TUTH), Nepal from January 2017 to December 2017. A total of 177 isolated from hospitalized patients were included in the study. The AST was performed by disc diffusion method. The MDR strains were identified by the criteria of Magiorakos et al, ESBL production by CLSI guidelines, and AmpC β-lactamase production by the AmpC disc test. MBL and KPC production were detected as per the method of Tsakris et al.

Results: Out of 177 , 91.0% were MDR isolates. Among the MDR isolates, the majority were isolated from respiratory tract specimens and were isolated from ICU patients. Most of the MDR isolates were resistant to all first-line antibiotics and all were completely sensitive to only polymyxin B and colistin sulfate. MBL (67.7%) was the common β-lactamase production among MDR isolates.

Conclusion: can cause a vast variety of infections in hospitalized patients. The highly resistant MDR strains are common in tertiary care hospitals. This bacteria lead to high morbidity and mortality as we are left with the only option of treating them by potentially toxic antibiotics like colistin sulfate and polymyxin B. Detection of drug resistance and rational use of antibiotics play a crucial role in the fight against this MDR pathogen.
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http://dx.doi.org/10.2147/IDR.S239514DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7061726PMC
March 2020

Carriage of Extended-Spectrum--Lactamase- and AmpC--Lactamase-Producing (ESBL-PE) in Healthy Community and Outpatient Department (OPD) Patients in Nepal.

Can J Infect Dis Med Microbiol 2020 11;2020:5154217. Epub 2020 Feb 11.

Department of Microbiology, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Extended-spectrum -lactamase (ESBL)- and AmpC--lactamase (ESBL)- and AmpC- have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC--lactamase (ESBL)- and AmpC- have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC.

Methods: During a 6-month period (November 2014-April 2015), a total of 190 stool specimens from 190 participants were obtained from different population. Of the total 260 fecal isolates, 152 from outpatient department (OPD) and 108 from healthy volunteer were collected. Stool specimens were cultured and enterobacterial isolates were subjected to antimicrobial susceptibility tests according to the standard microbiologic guidelines. ESBL was screened using ceftazidime (CAZ, 30 g) and cefotaxime (CTX, 30 g) and cefotaxime (CTX, 30 -lactamase (ESBL)- and AmpC.

Results: The prevalence of ESBL, AmpC--lactamase (ESBL)- and AmpC--lactamase (ESBL)- and AmpC- have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC- was 70.2% followed by (12.7%), and among AmpC--lactamase (ESBL)- and AmpC- was 70.2% followed by was 70.2% followed by (12.7%), and among AmpC- (12.7%), and among AmpC- 2/7 (28.57%) were detected highest among AmpC--lactamase (ESBL)- and AmpC.

Conclusion: Our study revealed a high prevalence of ESBL- and AmpC--lactamase-producing enteric pathogen in Nepalese OPD and healthy population. The significant increase of these isolates and increased rate of drug resistance indicates a serious threat that stress the need to implement the surveillance system and a proper control measure so as to limit the spread of ESBL-producing (ESBL-PE) in both OPD as well as in community. Therefore, healthcare providers need to be aware that ESBL- and AmpC--lactamase-producing strains are not only circulating in hospital environments but also in the community and should be dealt with accordingly.-lactamase (ESBL)- and AmpC- have recently emerged as a public threat in the treatment of nosocomial as well as community-acquired infections. Very little information is currently available about its existence in Nepal. We, therefore, aim to determine the prevalence of ESBL and AmpC--lactamase (ESBL)- and AmpC.
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http://dx.doi.org/10.1155/2020/5154217DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7036101PMC
February 2020

Recent advances in infection: focus on vaccine development.

Infect Drug Resist 2019 13;12:1243-1255. Epub 2019 May 13.

Department of Microbiology, Faculty of Medicine, Zagazig University, Zagazig, Egypt.

normally colonizes the nasal cavity and pharynx. After breaching the normal habitat, the organism is able to cause a number of infections at any site of the body. The development of antibiotic resistance has created a global challenge for treating infections. Therefore, protection by vaccines may provide valuable measures. Currently, several vaccine candidates have been prepared which are either in preclinical phase or in early clinical phase, whereas several candidates have failed to show a protective efficacy in human subjects. Approaches have also been made in the development of monoclonal or polyclonal antibodies for passive immunization to protect from infections. Therefore, in this review we have summarized the findings of recently published scientific literature to make a concise report.
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http://dx.doi.org/10.2147/IDR.S175014DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526327PMC
May 2019

Isolation, speciation and antifungal susceptibility testing of Candida isolates from various clinical specimens at a tertiary care hospital, Nepal.

BMC Res Notes 2017 Jun 24;10(1):218. Epub 2017 Jun 24.

Department of Microbiology, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Candida species are responsible for various clinical infections ranging from mucocutaneous infection to life threatening invasive diseases along with increased resistance to antifungal drugs has made a serious concern. Resistance to antifungal agents has increased during the last decade. Thus, identification of Candida up to species level and its antifungal susceptibility testing has a paramount significance in the management of Candidal infections. The aim of the study was to speciate Candida species and to determine antifungal susceptibility pattern of Candida species to antifungal agents.

Methods: A total of 100 consecutive Candida species were isolated from 1248 clinical specimens over 7 months period. Growths on Sabouraud dextrose agar were evaluated for colony appearance, macroscopic examination, Gram staining, germ tube test and urea hydrolysis test. Further, they were processed for Candida speciation on CHROMagar. Antifungal susceptibility testing was performed as recommended by Clinical and Laboratory Standards Institute (CLSI) M44-A document.

Results: Out of 100 Candida isolates, Candida albicans (56%) was the most common species. Among the non-albicans Candida species, Candida tropicalis (20%) was the predominant isolate followed by Candida glabrata (14%). Regarding antifungal susceptibility pattern, Candida species were more susceptible to clotrimazole (82%) followed by fluconazole (64%) and miconazole (44%).

Conclusions: Candida albicans was the predominant species responsible for various Candidal infections. Among commonly used antifungal drugs clotrimazole, miconazole and fluconazole were most effective.
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http://dx.doi.org/10.1186/s13104-017-2547-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5483268PMC
June 2017

High burden of antimicrobial resistance among gram negative bacteria causing healthcare associated infections in a critical care unit of Nepal.

Antimicrob Resist Infect Control 2017 15;6:67. Epub 2017 Jun 15.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Healthcare associated infections (HCAI) and antimicrobial resistance are principal threats to the patients of intensive care units and are the major determining factors for patient outcome. They are associated with increased morbidity, mortality, excess hospitalization and financial costs. The present study is an attempt to investigate the spectrum and antimicrobial resistance of bacterial isolates involved in healthcare associated infections (HCAI) in the patients of a critical care unit at a tertiary care university hospital in Kathmandu, Nepal.

Methods: A laboratory based study was conducted over the period of 15 months (January 2014 to March 2015) among the patients of intensive care unit of Tribhuvan University Teaching Hospital, Kathmandu, Nepal. Clinical specimens from patients with suspected healthcare-associated infection were processed and bacterial isolates were identified with standard microbiological methods. Antimicrobial susceptibilities of the isolated strains were determined according to the CLSI guidelines and β-lactamases (ESBL, AmpC, MBL and KPC) were detected by various phenotypic tests.

Results: One hundred and forty nine clinical specimens received from 135 patients suspected of HCAI (out of 491 patients) were found with significant bacterial growth. Specimens were from patients suspected of hospital-acquired pneumonia (16%, 79/491), bloodstream infections (5.7%, 28/491), surgical site infections (4.7%, 23/491), and urinary tract infections (3.9%, 19/491). spp., spp., and were the leading bacterial pathogens. Extremely high level of drug resistance (95.8%) along with the production of β-lactamases (ESBL; 43.7%, AmpC; 27.5%), MBL; 50.2% and KPC; 4.2%) was observed among Gram negative bacterial isolates.

Conclusion: Healthcare associated infections are very common in our ICU. Gram negative bacterial pathogens are major culprits associated with these infections and there is alarming state of drug resistance among these isolates. Continuous surveillance and establishment of preventive and control measures of healthcare associated infections are urgently needed in our setting.
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http://dx.doi.org/10.1186/s13756-017-0222-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5472869PMC
June 2017

Antimicrobial susceptibility pattern of Shigella spp. isolated from children under 5 years of age attending tertiary care hospitals, Nepal along with first finding of ESBL-production.

BMC Res Notes 2017 Jun 5;10(1):192. Epub 2017 Jun 5.

Department of Microbiology, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Shigella is an important cause of bacterial gastroenteritis in resource-poor countries. The treatment of shigellosis mostly requires antibiotics. However, the increase of multidrug resistance along with emergence of extended-spectrum β-lactamase and ciprofloxacin resistance among Shigella spp. has challenged the situation. This study was conducted to determine the distribution of species and antibiotic susceptibility pattern of Shigella species isolated from stool specimen among children less than 5 years of age in Nepal.

Results: Out of total 717 stool samples collected, 15 cases of Shigella spp. was isolated which includes 12 S. flexneri and 3 S. sonnei. Multidrug resistance was found among 13(86%) of the isolates. One of the isolates of S. flexneri was found to be ESBL-producer with MIC >256 mg/L for cefixime.

Conclusion: The high occurrence of multidrug resistance among Shigella spp. along with a case of ESBL-production for the first time in Nepal alarms the concerns about dissemination of the resistant isolates. So, systemic monitoring of the antimicrobial susceptibility pattern of Shigella spp. is becoming crucial to guide therapy.
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http://dx.doi.org/10.1186/s13104-017-2512-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5460519PMC
June 2017

Epidemiology of device-associated infections in an intensive care unit of a teaching hospital in Nepal: A prospective surveillance study from a developing country.

Am J Infect Control 2017 Sep 18;45(9):1024-1029. Epub 2017 Apr 18.

Department of Clinical Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Device-associated health care-acquired infections (DA-HAIs) in intensive care unit patients are a major cause of morbidity, mortality, and increased health care costs.

Methods: A prospective, structured clinicomicrobiological surveillance was carried out for 3 common DA-HAIs: ventilator-associated pneumonia (VAP), central line-associated bloodstream infection (CLABSI), and catheter-associated urinary tract infection (CAUTI) present in the patients of an intensive care unit of a teaching hospital in Nepal. DA-HAIs were identified using the Centers for Disease Control and Prevention definitions, and their rates were expressed as number of DA-HAIs per 1,000 device-days.

Results: Overall incidence rate of DA-HAIs was 27.3 per 1,000 patient-days occurring in 37.1% of patients. The device utilization ratio for mechanical ventilation, central line catheter, and urinary catheter was 0.83, 0.63, and 0.78, respectively. The rates of VAP, CLABSI, and CAUTI were 21.40, 8.64, and 5.11 per 1,000 device-days, respectively. Acinetobacter spp (32.7%), Klebsiella spp (23.6%), Burkholderia cepacia complex (12.7%), and Escherichia coli (10.9%) were the common bacterial pathogens. Most of the bacterial isolates associated with DA-HAIs were found to be multidrug-resistant.

Conclusions: Incidence of DA-HAIs in the study intensive care unit was high compared with that of developed countries. Formulation and implementation of standard infection control protocols, active surveillance of DA-HAIs, and antimicrobial stewardship are urgently needed in our country.
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http://dx.doi.org/10.1016/j.ajic.2017.02.040DOI Listing
September 2017

Molecular epidemiology of Rotavirus causing diarrhea among children less than five years of age visiting national level children hospitals, Nepal.

BMC Pediatr 2017 04 7;17(1):101. Epub 2017 Apr 7.

Department of Microbiology, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Background: Rotaviruses are the major cause of diarrhea among the infants and young children all over the world causing over 500,000 deaths and 2.4 million hospitalizations each year. In Nepal Rotavirus infection positivity rates ranges from 17.0 to 39.0% among children less than 5 years. However, little is known about the molecular genotypes of Rotavirus prevailing. The objective of this study was to estimate the burden of Rotavirus gastroenteritis and determine the genotypes of Rotavirus among children less than 5 years.

Methods: The cross sectional study was conducted from January to November 2014 among children less than 5 years old visiting Kanti Children's Hospital and Tribhuvan University Teaching Hospital. Rotavirus antigen detection was performed by Enzyme Linked Immunosorbent Assay (ELISA) using ProSpecT Rotavirus Microplate Assay. Among the Rotavirus antigen positive samples, 59 samples were used for Rotavirus RNA extraction. Multiplex PCR was performed to identify G type comprising G1-G4, G8-G10 and G12 and P type comprising P[4], P[6], P[8], P[9], P[10], and P[11].

Results: A total of 717 diarrheal stool samples were collected from patients ranging from 10 days to 59 months of age. Rotavirus antigen positive was found among (N = 164)22.9% of patients. The highest number of the diarrhea was seen in January. Molecular analysis of Rotavirus genotypes revealed that the predominant G-Type was G12 (36%) followed by G9 (31%), G1 (21%), G2 (8.6%). The predominant P- type was P6 (32.8%) followed by P8 (31%), P10 (14.8%), P4 (14.8%). A total of seven G/P type combinations were identified the most common being G12P [6] (35.8%), G1P [8] (15.1%), G9P [8] (15.1%).

Conclusion: Rotavirus diarrhea is, mostly affecting children from 7 to 24 months in Nepal, mostly occurring in winter. The circulating genotypes in the country are found to be primarily unusual genotypes and predominance of G12P[6]. It is recommended to conduct genotyping of Rotavirus on large samples before starting vaccination in the country.
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http://dx.doi.org/10.1186/s12887-017-0858-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5385007PMC
April 2017

Ecrg4 contributes to the anti-glioma immunosurveillance through type-I interferon signaling.

Oncoimmunology 2016;5(12):e1242547. Epub 2016 Oct 14.

Division of Stem Cell Biology, Institute for Genetic Medicine, Hokkaido University , Sapporo, Hokkaido, Japan.

Esophageal cancer-related gene 4 (), a hormone-like peptide, is thought to be a tumor suppressor, however, little is known about the mechanism of how Ecrg4 suppresses tumorigenesis. Here, we show that the null glioma-initiating cell (GIC) line, which was generated from neural stem cells of knockout (KO) mice, effectively formed tumors in the brains of immunocompetent mice, whereas the transplanted wild type-GIC line GIC(+/+) was frequently eliminated. This was caused by host immune system including adaptive T cell responses, since depletion of CD4, CD8, or NK cells by specific antibodies recovered tumorigenicity of GIC(+/+). We demonstrate that Ecrg4 fragments, amino acid residues 71-132 and 133-148, which are produced by the proteolitic cleavage, induced the expression of pro-inflammatory cytokines in microglia . Moreover, blockades of type-I interferon (IFN) signaling , either depleting IFN-α/β receptor 1 or using KO mice, abrogated the Ecrg4-dependent antitumor activity. Together, our findings indicate a major antitumor function of Ecrg4 in enhancing host immunity via type-I IFN signaling, and suggest its potential as a clinical candidate for cancer immunotherapy.
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http://dx.doi.org/10.1080/2162402X.2016.1242547DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5214586PMC
October 2016

Clinicomycological Characterization of Superficial Mycoses from a Tertiary Care Hospital in Nepal.

Dermatol Res Pract 2016 24;2016:9509705. Epub 2016 Nov 24.

Department of Microbiology, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

. Superficial mycosis is a common fungal infection worldwide, mainly caused by dermatophytes. However, the prevalence of species varies geographically. In addition, fungal treatment is best guided according to species isolated. This study was carried out to determine the clinical as well as mycological profile of superficial mycoses in a tertiary care hospital, Nepal. . This was a prospective case-control laboratory based study conducted over a period of six months from January to June 2014 at Tribhuvan University Teaching Hospital, Nepal. A total of 200 specimens were collected from the patients suspected of superficial mycoses. The specimens were macroscopically as well as microscopically examined. The growth was observed up to 4 weeks. Out of total 200 specimens from the patients suspected of superficial mycoses, tinea corporis 50 (25%) was most common clinical types. KOH mount was positive in 89 (44.5%) and culture was positive in 111 (55.5%). 44 (39.6%) was the most common isolate. The diagnostic yields of KOH mount and culture were found to be complementary to each other. Thus both the methods added with clinical findings are equally important to establish superficial mycosis.
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http://dx.doi.org/10.1155/2016/9509705DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5143727PMC
November 2016

Detection of biofilm production and antibiotic resistance pattern in clinical isolates from indwelling medical devices.

Curr Microbiol 2015 Jan 20;70(1):128-34. Epub 2014 Sep 20.

Department of Microbiology, Institute of Medicine, Tribhuvan University, Maharajgunj, Kathmandu, Nepal,

Microbial biofilms pose great threat for patients requiring indwelling medical devices (IMDs) as it is difficult to remove them. It is, therefore, crucial to follow an appropriate method for the detection of biofilms. The present study focuses on detection of biofilm formation among the isolates from IMDs. We also aimed to explore the antibiogram of biofilm producers. This prospective analysis included 65 prosthetic samples. After isolation and identification of bacteria following standard methodology, antibiogram of the isolates were produced following Kirby-Bauer disc diffusion method. Detection of biofilms was done by tube adherence (TA), Congo red agar and tissue culture plate (TCP) methods. Out of 67 clinical isolates from IMDs, TCP detected 31 (46.3 %) biofilm producers and 36 (53.7 %) biofilm non-producers. Klebsiella pneumoniae, Pseudomonas aeruginosa and Burkholderia cepacia complex were found to be the most frequent biofilm producers. The TA method correlated well with the TCP method for biofilm detection. Higher antibiotic resistance was observed in biofilm producers than in biofilm non-producers. The most effective antibiotics for biofilm producing Gram-positive isolates were Vancomycin and Tigecycline, and that for biofilm producing Gram-negative isolates were Polymyxin-B, Colistin Sulphate and Tigecycline. Nearly 46 % of the isolates were found to be biofilm producers. The antibiotic susceptibility pattern in the present study showed Amoxicillin to be an ineffective drug for isolates from the IMDs. For the detection of biofilm production, TA method can be an economical and effective alternative to TCP method.
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http://dx.doi.org/10.1007/s00284-014-0694-5DOI Listing
January 2015

Emerging threat of multidrug resistant bugs--Acinetobacter calcoaceticus baumannii complex and methicillin resistant Staphylococcus aureus.

BMC Res Notes 2013 Mar 15;6:98. Epub 2013 Mar 15.

Department of Microbiology, Institute of Medicine, Tribhuvan University, Kathmandu, Nepal.

Background: Infections caused by bacteria such as multidrug resistant (MDR) Acinetobacter spp. and methicillin-resistant Staphylococcus aureus (MRSA) constitute a worldwide pandemic. Without gathering information about these strains, we cannot reduce the morbidity and mortality due to infections caused by these notorious bugs.

Methods: This study was conducted to identify the status of MDR Acinetobacter spp. and MRSA in a tertiary care centre of Nepal. Sputum, endotracheal aspirate and bronchial washing specimens were collected and processed from patients suspected of lower respiratory tract infection following standard microbiological methods recommended by the American Society for Microbiology (ASM). Double disk synergy test method was employed for the detection of extended-spectrum beta-lactamase (ESBL) in Acinetobacter isolates. Methicillin resistance in S. aureus was confirmed by using cefoxitin and oxacillin disks.

Results: Different genomespecies of Acinetobacter were isolated; these consisted of Acinetobacter calcoaceticus baumannii complex and A. lwoffii. Around 95% of Acinetobacter isolates were MDR, while 12.9% were ESBL-producer. Of the total 33 isolates of S. aureus, 26 (78.8%) were MDR and 14 (42.4%) were methicillin resistant.

Conclusions: A large number of MDR Acinetobacter spp. and MRSA has been noted in this study. The condition is worsened by the emergence of ESBL producing Acinetobacter spp. Hence, judicious use of antimicrobials is mandatory in clinical settings. Moreover, there should be vigilant surveillance of resistant clones in laboratories.
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http://dx.doi.org/10.1186/1756-0500-6-98DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3605284PMC
March 2013

Characterization of rotavirus causing acute diarrhoea in children in Kathmandu, Nepal, showing the dominance of serotype G12.

J Med Microbiol 2013 Jan 4;62(Pt 1):114-120. Epub 2012 Oct 4.

Department of Microbiology, Institute of Medicine, Tribhuvan University Teaching Hospital, Kathmandu, Nepal.

Diarrhoeal diseases are a major problem in developing countries. Though precise data on childhood mortality associated with diarrhoeal diseases in Nepal are not available, it has been estimated that approximately 25 % of child deaths are associated with diarrhoeal disease, particularly acute diarrhoea. The purpose of this study was to assess the incidence of rotavirus causing acute diarrhoea in children less than 5 years of age. A total of 525 children with acute diarrhoea in a children's hospital of Kathmandu, Nepal, were enrolled between April and September 2011. The incidence of acute diarrhoea due to rotavirus was 25.9 % (136/525) as determined by ELISA. The percentage of rotavirus-infected males was higher (64.5 %) than females (35.5 %). The frequency of rotavirus cases was higher in children less than 2 years of age, among which the majority of cases (80.2 %) were in children between 6 and 24 months old (P<0.01). Genotypic characterization by RT-PCR revealed that the serotype G12 represented 55.9 % of cases in this study associated with P-types of either P[6], P[4] or P[8]. Further to this, a total of eight G/P combinations were identified, G12P[6] being the most common strain type of rotavirus in Nepal, with a prevalence rate of 46.4 %. The aim of this study was to find out the major genotypes of rotavirus causing acute diarrhoea in children.
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http://dx.doi.org/10.1099/jmm.0.048124-0DOI Listing
January 2013
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