Infective Vaginal Discharge among Women of the Reproductive Age in the Outpatient Department of a Primary Care Centre.

Introduction: Infective vaginal discharge is the most common complaint of the women of reproductive age group attending gynaecology outpatient department. Vaginal discharge may be normal or abnormal. Infective vaginal discharge is usually related to one of the three conditions, like bacterial vaginosis, vulvovaginal candidiasis, and trichomoniasis. This study aimed to find out the prevalence of infective vaginal discharge among women of the reproductive age in the outpatient department of a primary care centre. Methods: A descriptive cross-sectional study was carried out among women of the reproductive age group visiting the outpatient Department of the primary care hospital from 1 March 2022 to 1 August 2022 after obtaining ethical approval from the Institutional Review Committee. A convenience sampling method was used. The point estimate was calculated at a 95% Confidence Interval. Results: Among 138 patients, infective vaginal discharge was found in 42 (30.43%) (21.32-39.54, 95% Confidence Interval). Bacterial vaginosis was seen in 22 (52.38%), Candida was seen in 13 (30.95%), and Trichomonas was seen in 7 (16.66%) cases. Conclusions: The prevalence of infective vaginal discharge was lower as compared to other studies done in similar settings. Keywords: bacterial; candidiasis; prevalence; vaginitis.

Acute Conjunctivitis among Patients Visiting the Outpatient Department of Ophthalmology in a Tertiary Care Centre.

Introduction: Conjunctivitis is a highly prevalent ocular disease that flares up every year. The humidity and high temperature favour the causative agents responsible for the epidemic. Acute infective conjunctivitis may be either viral or bacterial, a distinct type of condition with unique clinical features and treatment approaches. The aim of the study was to find out the prevalence of acute conjunctivitis among patients visiting the outpatient Department of Ophthalmology in a tertiary care centre. Methods: This descriptive cross-sectional study was conducted among the patients visiting the outpatient Department of Ophthalmology. Data of 30 August 2023 to 30 September 2023 was collected between 21 November 2023 to 24 November 2023. All patients presenting in the Ophthalmology Department having complete hospital record were included in the study. Patients having missing data on the medical records of the hospital were excluded. A convenience sampling method was used. The point estimate was calculated at a 95% Confidence Interval. Results: Among 5,507 patients, acute conjunctivitis was seen in 1240 (22.52%) (21.42-23.62, 95% Confidence Interval). The majority were male 732 (59.03%) and adults 760 (61.29%) with a mean age of 32.56±18.74 years. Conclusions: The prevalence of conjunctivitis among patients visiting the outpatient Department of Opthalmology was found to be higher than other studies done in similar settings. Keywords: conjunctivitis; disease outbreaks; enterovirus.

Multidrug Resistant Escherichia coli among Urinary Samples of Patients with Urinary Tract Infection in the Microbiology Department of a Tertiary Care Center: A Descriptive Cross-Sectional Study.

Introduction: Urinary Tract Infection one of the most common and manageable infections still holds its position as a major public health issue worldwide due to an increase in the number of multidrug resistant bacteria. This study aims to find out the prevalence of multidrug resistant Escherichia coli among urinary samples of patients with urinary tract infections in the microbiology Department of a tertiary care center. Methods: A descriptive cross-sectional study was carried out at a tertiary care centre from 8 August 2018 to 9 January 2019. Ethical approval was received from the Institutional Review Committee (Reference number: 123/2018). Clinically suspected cases of urinary tract infection were included in this study. A convenience sampling method was used. Point estimate and 95% Confidence Interval were calculated. Results: Among 594 patients with urinary tract infections, the prevalence of multidrug resistant Escherichia coli was 102 (17.17%) (14.14-20.20, 95% Confidence Interval). Out of which, the production of extended-spectrum beta-lactamase and AmpC beta-lactamase were observed in 74 (72.54%), and 28 (27.45%) isolates respectively. The co-production of extended-spectrum beta-lactamases/AmpC was observed in 17 (16.67%). Conclusions: The prevalence of multidrug resistant Escherichia coli among patients urinary samples of patient with urinary tract infection was lower as compared to the other studies done in similar settings. Keywords: antibiotics; Escherichia coli; urinary tract infection.

Phenotypical Detection of ß-Lactamases in a Multidrug-Resistant and Extensively Drug-Resistant Chryseobacterium indologens: A Rare Human Pathogen With Special References to Risk Factor.

Chryseobacterium indologenes is gram-negative bacteria that cause infection in humans. It is less frequently isolated in the laboratory. The development of drug-resistant and its intrinsic ability to resist a wide range of antimicrobials enables them to cause mortality in an immunocompromised patient with a longer hospital stay. Our study objectives are to investigate antimicrobial-resistant patterns, drug-resistant enzymes, and the risk factor analysis associated with multidrug-resistant (MDR), extensively drug-resistant (XDR), and Pan-drug resistant (PDR) within 2 years. Altogether 53 strains of Chryseobacterium indologens were obtained from 5000 specimens that were processed for routine bacterial culture. The bacterial identification was done using conventional techniques (colony morphology, gram staining, flexirubin test, and biochemical tests) as well as the VITEK-2 System to further confirm. The bacterial isolate were processed to observe antimicrobial susceptibility test (AST) using disk diffusion method. MDR XDR and PDR were classified following European Centre for Disease Prevention and Control guidelines. C. indologens strains with beta-lactamases such as extended-spectrum beta-lactamases (ESBL), metallo beta-lactamases (MBL), and Amp-C beta-lactamases (Amp-C) were detected phenotypically. The highest isolation of C. indologens was observed in a sputum sample. In vitro antimicrobial susceptibility test revealed susceptibility to tigecycline followed by levofloxacin, cotrimoxazole, and piperacillin-tazobactam. From 53 isolates of C. indologens, MDR accounts for 56.60% and 22.64% for XDR. Combined antimicrobial therapy and longer hospital stay were found to be the leading risk factor. All 53 C. indologenes strains were detected as MBL. Total ESBL was detected in 16.98% of MBL producer strains and Amp-C was observed in 13.20% of MBL-producing strains. All 3 enzyme co-oproducers were seen in only 5.66% of C. indologens. Although it is rarely encountered in the laboratory, it showed a remarkable effect in patients with underlying predisposing factors and prolonged hospital stays. The presence of betalactamases determined the drug-resistant activity on a wide spectrum of tested antibiotics.

Multidrug-resistant among Non-Fermenting Gram-negative Bacteria Isolated in the Department of Microbiology of a Tertiary Care Centre.

Introduction: Infection caused by Non-fermenting Gram-negative bacteria (NFGNB) like Pseudomonas aeruginosa and Acinetobacter baumannii leads to life-threatening conditions. These bacteria are often multidrug-resistant which leads to limited therapeutic options leading to treatment failure. Little information is available regarding the prevalence and resistance pattern of such bacteria in our country. The aim of the study was to find out the prevalence of multidrug-resistant among non-fermenting Gram-negative bacteria isolated in the Department of Microbiology of a tertiary care centre. Methods: A descriptive cross-sectional study was conducted in the Department of Microbiology of a tertiary care centre from 1 September 2021 to 30 August 2022 after obtaining ethical approval from the Institutional Review Committee. All samples received in the Microbiology laboratory for diagnostic purposes were included. A convenience sampling method was used. The point estimated was calculated at a 95% Confidence Interval. Results: Among 412 non-fermenting Gram-negative bacteria, multidrug resistance was observed in 373 (90.53%) (87.70-93.36, 95% Confidence Interval) isolates. Among 373 isolates, Acinetobacter baumannii was 253 (67.83%) and Pseudomonas aeruginosa was 120 (32.17%). Conclusions: The prevalence of multidrug-resistant non-fermenting Gram-negative bacteria was found to be higher than in the study conducted in similar settings. Keywords: Acinetobacter baumannii; antibiotic; Pseudomonas aeruginosa; prevalence.

Comparative genomic analyses of multi-drug resistant Mycobacterium tuberculosis from Nepal and other geographical locations.

Nepal exhibits a tuberculosis (TB) incidence rate that is comparable to neighbouring high TB incidence countries. In addition, it records >500 cases of multi-drug resistant (MDR) TB each year. The objective of this study was to perform whole-genome bioinformatic analysis on MDR-TB isolates from Nepal (n = 19) to identify the specific mutations underlying their phenotypic resistance. In addition, we examined the dominant genotype among the Nepal MDR-TB isolates, the East-Asian Beijing sub-lineage, to determine its relatedness to a panel of 1274 genomes of international strains available from public databases. These analyses provided evidence that the XDR-TB isolates in our collection were not derived from importation of primary XDR-TB to Nepal but were more likely the result of acquisition of second-line drug resistance in Nepal. Resistance to fluoroquinolones was detected among a high proportion of the Nepal isolates. This has implications for the management of TB, including appropriate antimicrobial stewardship and susceptibility testing for fluoroquinolones and other second-line TB drugs, to minimise the development of XDR-TB among Nepal TB cases.

Diagnostic performance of GeneXpert MTB/RIF assay compared to conventional Mycobacterium tuberculosis culture for diagnosis of pulmonary and extrapulmonary tuberculosis, Nepal.

Tuberculosis is an infectious disease caused by the Mycobacterium tuberculosis. It is a global health problem and major cause of death in resource-limited countries like Nepal. Timely diagnosis with sensitive testing methods could assist in early management of the disease. This study was conducted to compare the diagnostic performance of GeneXpert MTB/RIF and conventional acid-fast staining with M. tuberculosis culture. The study was carried out in the Department of Microbiology, Shree Birendra Army Hospital, Nepal. Samples (n=500) were tested with a GeneXpert MTB/RIF assay and acid-fast bacilli (AFB) smear microscopy. All samples were sent for M. tuberculosis conventional culture by the German-Nepal Tuberculosis Project, Kathmandu, Nepal (GENETUP). Out of a total 500 pulmonary and extrapulmonary samples tested, 97 samples were positive for M. tuberculosis by GeneXpert MTB/RIF assay. Out of the positive samples, only 95 samples were found positive by the culture method. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of AFB microscopy was 45.3%, 99.5%, 99.5% and 88.5%, respectively. The sensitivity, specificity, PPV and NPV of GeneXpert MTB/RIF was found to be 100%, 99.5%, 97.5% and 100%, respectively compared to the gold standard culture method. The GeneXpert MTB/RIF test was comparable with culture diagnosis of both pulmonary and extrapulmonary tuberculosis cases.

Draft Genome Sequence of an Isolate of Extensively Drug-Resistant Mycobacterium tuberculosis from Nepal.

Extensively drug-resistant (XDR) Mycobacterium tuberculosis has become a challenge to the treatment of tuberculosis (TB) in several countries, including Nepal. Here, we report for the first time the draft genome sequence of an isolate of XDR-TB collected in Nepal and describe single-nucleotide variations associated with its extensively drug-resistant phenotype.

Biofilm and metallo beta-lactamase production among the strains of Pseudomonas aeruginosa and Acinetobacter spp. at a Tertiary Care Hospital in Kathmandu, Nepal.

INTRODUCTION: Pseudomonas aeruginosa and Acinetobacter spp. are found to be associated with biofilm and metallo-ß-lactamase production and are the common causes of serious infections mainly in hospitalized patients. So, the main aims of this study were to determine the rates of biofilm production and metallo beta-lactamase production (MBL) among the strains of Pseudomonas aeruginosa and Acinetobacter spp. isolated from hospitalized patients. METHODS: A total of 85 P. aeruginosa isolates and 50 Acinetobacter spp. isolates isolated from different clinical specimens from patients admitted to Shree Birendra Hospital, Kathmandu, Nepal from July 2013 to May 2014 were included in this study. The bacterial isolates were identified with the help of biochemical tests. Modified Kirby-Bauer disc diffusion technique was used for antimicrobial susceptibility testing. Combined disc diffusion technique was used for the detection of MBL production, while Congo red agar method and tube adherence method were used for detection of biofilm production. RESULTS: Around 16.4% of P. aeruginosa isolates and 22% of the strains of Acinetobacter spp. were metallo ß-lactamase producers. Out of 85 P. aeruginosa isolates, 23 (27.05%) were biofilm producers according to tube adherence test while, only 13 (15.29%) were biofilm producers as per Congo red agar method. Similarly, out of 50 Acinetobacter spp. 7 (14%) isolates were biofilm producers on the basis of tube adherence test, while only 5 (10%) were positive for biofilm production by Congo red agar method. Highest rates of susceptibility of P. aeruginosa as well as Acinetobacter spp. were seen toward colistin. CONCLUSION: In our study, biofilm production and metallo beta-lactamase production were observed among Pseudomonas aeruginosa and Acinetobacter spp. However, no statistically significant association could be established between biofilm production and metallo beta-lactamase production.

Detection of Methicillin Resistant Staphylococcus aureus and Determination of Minimum Inhibitory Concentration of Vancomycin for Staphylococcus aureus Isolated from Pus/Wound Swab Samples of the Patients Attending a Tertiary Care Hospital in Kathmandu, Nepal.

The present study was conducted to evaluate the performance of cefoxitin disc diffusion method and oxacillin broth microdilution method for detection of methicillin resistant S. aureus (MRSA), taking presence of mecA gene as reference. In addition, inducible clindamycin resistance and beta-lactamase production were studied and minimum inhibitory concentration (MIC) of vancomycin for S. aureus isolates was determined. A total of 711 nonrepeated pus/wound swab samples from different anatomic locations were included in the study. The Staphylococcus aureus was identified on the basis of colony morphology, Gram's stain, and biochemical tests. A total of 110 (15.47%) S. aureus isolates were recovered, of which 39 (35.50%) isolates were identified as MRSA by cefoxitin disc diffusion method. By oxacillin broth microdilution method, 31.82% of the Staphylococcus aureus isolates were found to be MRSA. However, mecA gene was present in only 29.1% of the isolates. Further, beta-lactamase production was observed in 71.82% of the isolates, while inducible clindamycin resistance was found in 10% of S. aureus isolates. The MIC value of vancomycin for S. aureus ranged from 0.016 µg/mL to 1 µg/mL. On the basis of the absolute sensitivity (100%), both phenotypic methods could be employed for routine diagnosis of MRSA in clinical microbiology laboratory; however cefoxitin disc diffusion could be preferred over MIC method considering time and labour factor.

Correlation between biofilm formation and resistance toward different commonly used antibiotics along with extended spectrum beta lactamase production in uropathogenic Escherichia coli isolated from the patients suspected of urinary tract infections visiting Shree Birendra Hospital, Chhauni, Kathmandu, Nepal.

BACKGROUND: Escherichia coli is the most predominant causative agent of urinary tract infection (UTI). Recently, increase in drug resistance among the uropathogenic bacteria has caused great problem in treatment of UTI. The main objective of this research is to determine the correlation between biofilm formation and resistance toward different commonly used antibiotics along with extended spectrum beta lactamase production in uropathogenic Escherichia coli. METHODS: The urine samples collected from the patients suspected of urinary tract infections (visiting Shree Birendra Hospital, Chhauni, Kathmandu, Nepal between July to December 2013) were cultured in cystine lactose electrolyte deficient (CLED) agar by using semi quantitative culture technique. Extended spectrum beta lactamase (ESBL) production was detected by combined disc diffusion technique and biofilm formation was detected by Congo red agar method. Chi-square test was applied and p-value < 0.05 was considered statistically significant. RESULTS: Out of 1480 urine samples, E. Coli was isolated from 208 (14.1 %) samples. Of total 69 (33.2 %) ESBL producing uropathogenic strains of E. coli, 20 (29 %) were strong biofilm producers, 22 (31.9 %) were moderate biofilm producers, 11 (15.9 %) were weak biofilm producers and 16 (23.2 %) were biofilm non producers. Whereas among 139 ESBL non producing E. coli, 22 (15.8 %) were strong biofilm producers, 20 (14.4 %) were moderate biofilm producers, 13 (9.4 %) were weak biofilm producers and 84 (60.4 %) were biofilm non producers. Among total 108 biofilm producing E. coli, maximum resistance was observed toward cephalexin followed by amoxicillin and highest susceptibility was seen toward amikacin. CONCLUSION: The ability of biofilm formation was found to be significantly higher in ESBL producing strains of E. coli than that in ESBL non producing strains (p < 0.05). There was higher resistance rate to antimicrobial agents among biofilm producing strains of E. coli than that in biofilm non producing strains. According to our antimicrobial susceptibility pattern for E. coli, to start preliminary treatment for UTI in Nepal, we recommend to use amikacin or nitrofurantoin. Further, for the treatment of the UTI, the antibiotics should be selected on the basis of the urine culture and sensitivity report.

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

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.