Tribhuvan University Journal of Microbiology 2021-02-11T19:43:23+00:00 Dr. Megha Raj Banjara Open Journal Systems <p>The Tribhuvan University Journal of Microbiology is published by the Central Department of Microbiology, Tribhuvan University, Kirtipur, Kathmandu, Nepal. Full text content available here and on the journal's own website <a href="" target="_blank" rel="noopener"></a></p> Contribution of Microbiologists in Nepal during COVID-19 Pandemic 2021-02-11T19:42:58+00:00 Megha Raj Banjara <p>No abstract available.</p> 2021-02-11T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Antifungal Susceptibility Pattern of Candida Isolates Causing Vulvovaginitis in Reproductive Age Women 2021-02-11T19:42:56+00:00 Pabitra Shrestha Sunil Mani Pokharel Anima Shrestha <p><strong>Objectives: </strong>The study was designed to isolate and identify <em>Candida </em>species from high vaginal swab, and to determine the antifungal susceptibility pattern of <em>Candida </em>spp. among women of reproductive age group i.e. 15- 50 years old.</p> <p><strong>Methods: </strong>High vaginal swabs were processed to isolate <em>Candida </em>species and identified by Gram’s stain, germ tube formation test, carbohydrate (glucose, sucrose, maltose, lactose) fermentation tests and antifungal susceptibility test were performed as recommended by Clinical Laboratory Standard Institute.</p> <p><strong>Results: </strong>Out of total 261 samples, 42.1% (110/261) were found to be culture positive for <em>Candida </em>spp. <em>Candida albicans </em>was the most common (56.4%) and among non-albicans, predominate species was <em>C. krusei </em>(19.1%) isolated from high vaginal swab specimens All the isolates of <em>Candida </em>species were sensitive to Nystatin and Miconazole and resistant to Itraconazole. Fluconazole, Itraconazole and Clotrimazole are widely used drug against vaginal candidiasis but showed high resistance which leads to treatment failure.</p> <p><strong>Conclusion: </strong>This study on the infection rate of <em>Candida </em>and its antifungal susceptibility pattern may help in the choice of appropriate therapy in the clinical setting.</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Antibacterial Activity of Common Spices Extracts on Bacterial Isolates found in Kachhila, a Newari Cuisine 2021-02-11T19:42:59+00:00 Shiv Nandan Sah Hemanta Khanal Dev Raj Acharya <p><strong>Objectives: </strong>The objective of this study was to determine the antibacterial property of aqueous extracts of common spices used in the preparation of <em>Kachhila </em>such as garlic, ginger, and turmeric.</p> <p><strong>Methods: </strong>After washing and cleaning separately, aqueous extracts were extracted from each spice by crushed with mortar and pestle. Antibacterial activity of aqueous extracts of each spice was evaluated by using agar well diffusion assay and paper disc diffusion assay against test bacteria isolated from buff meat used in <em>Kachhila </em>preparation<em>. </em></p> <p><strong>Results: </strong>Minimum inhibition concentration value of each spice was evaluated against all bacterial isolates. Total bacterial count (CFU/g) of prepared <em>Kachhila </em>was determined in each 30 mins interval up to 150 mins to reveal antibacterial activities of spices used in <em>Kachhila</em>. The antibacterial effect of aqueous extract of garlic extract was found to be the stronger in comparison, followed by turmeric and ginger against test bacterial isolates. The MIC of individual spice extract was found to be 125 to 4000 μl/ml against all the test bacteria. Spices used in <em>Kachhila </em>revealed that bacterial load decreased with time due to the antibacterial property of spices.</p> <p><strong>Conclusion: </strong>This study concluded that the antibacterial effect of aqueous extract of Garlic extract was stronger in comparison, followed by Turmeric and Ginger against four test bacteria isolated from buff meat namely; <em>E. coli, Salmonella </em>spp., <em>Pseudomonas </em>spp., <em>Staphylococcus aureus</em>. Therefore this study revealed that spices used in <em>Kachhila </em>have an antibacterial property and enhance the shelf life of <em>Kachhila.</em> &nbsp;</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Methicillin Resistant Staphylococcus aureus in Health Care Workers of a Tertiary Care Infectious Disease Hospital in Nepal 2021-02-11T19:43:00+00:00 Priyanka Shah Binod Dhungel Anup Bastola Megh Raj Banjara Komal Raj Rijal Prakash Ghimire <p><strong>Objectives: </strong>Acquisition of <em>mecA </em>gene in infectious strains of Methicillin resistant <em>Staphylococcus aureus </em>(MRSA) are considered as one of the potential virulence factors that enables the host bacteria to carry out several nosocomial and community-acquired infections. The main aim of this study was to determine the prevalence of MRSA, their antibiogram and <em>mec</em>A gene in the bacterial isolates obtained from the asymptomatic healthcare workers (HCWs) working in Sukraraj Tropical and Infectious Disease Hospital (STIDH), Kathmandu Nepal.</p> <p><strong>Methods: </strong>This prospective cross-sectional study involved the collection of nasal and hands swab of 125 randomly selected HCWs from December 2019 to February 2020. Conventional microbiological methods were used to isolate and identify <em>S. aureus</em>. Antimicrobial susceptibility testing was done by modified Kirby Bauer disc diffusion method. MRSA was confirmed by using cefoxitin disc. Detection of mecA gene in the chromosome which was extracted by Phenol: Chloroform: isoamyl alcohol DNA extraction method, amplified by using PCR and visualized by running agarose gel electrophoresis.</p> <p><strong>Results: </strong>The overall and MRSA carriage rate among the HCWs was found to be 28% (35/125) and 10.4% (13/125) respectively. <em>S. aureus </em>carriage rate was highest among sanitation staffs (34.2%) followed by pharmacy staffs (33.3%), laboratory personnel (18.8%), doctors (9.1%) and nurses (7.5%). Similarly, 34.2% (13/38) of the <em>S. aureus </em>isolates were resistant to methicillin, 31.6% (12/38) were inducible-clindamycin resistant and 63.2% (24/38) of them were multi-drug resistant (MDR). All the 13 MRSA isolates harbored the <em>mecA </em>gene.</p> <p><strong>Conclusions: </strong>Carriage rate of MRSA among HCWs was high and alarming, indicating the prompt need of intervention measures to curb the growth and spread of resistant isolates in the hospital settings. Effective surveillance (of infectious diseases) and establishment of advanced diagnostic facilities can assist in estimating the actual burden of the MRSA which in turn helps to formulate and implement the appropriate policies and infection-control programs to address the increasing antimicrobial resistance in the country. &nbsp;</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Antibiotic Susceptibility Pattern of Salmonella Enterica serovars Typhi and Paratyphi A Isolated From Patients Suspected of Enteric Fever 2021-02-11T19:43:02+00:00 Dhirendra Niroula Jyotsna Shrestha Supriya Sharma Anjana Singh <p><strong>Objectives: </strong>The study aimed to assess the antibiotic susceptibility profile of <em>Salmonella </em>spp isolated from patients suspected of enteric fever.</p> <p><strong>Methods: </strong>This cross-sectional prospective study was carried out from April to June, 2014among 484 patients clinically suspected of enteric fever visiting Bir Hospital, Kathmandu, Nepal. Blood sample collected from each patient was processed for culture in bile broth. Identification of <em>Salmonella </em>spp was done by conventional microbiological techniques including colony characteristics, Gram's staining and biochemical tests. Antibiotic susceptibility testing of identified isolates was done by Kirby-Bauer disk diffusion method following the 2014 CLS I guideline.</p> <p><strong>Results: </strong>Out of 484 blood samples, 36 (7.43%) cases showed the growth of <em>Salmonella </em>spp; of which 27 (75%) were <em>Salmonella enterica </em>serovar Typhi (ST) and 9 (25%) were <em>Salmonella enterica </em>Paratyphi A (SPA). Among the <em>Salmonella </em>isolates, 5.55% were multidrug resistant and 41.66% were fluoroquinolone resistant. More than 80% of isolates were sensitive to chloramphenicol, amoxicillin, and cotrimoxazole whereas 58%, 50% and 6% of isolates were sensitive to fluoroquinolone antibiotics i.e. ciprofloxacin, ofloxacin and nalidixic acid respectively. All the isolates were susceptible to ceftazidime. All SPA and 89% of ST were sensitive to azithromycin.</p> <p><strong>Conclusion: </strong>Higher percentage of susceptible isolates to chloramphenicol, cotrimoxazole, and amoxicillin suggests the reconsideration of these antibiotics for the treatment of enteric fever. Azithromycin can be considered as drug of choice for the treatment of enteric fever. &nbsp;</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Prevalence of Methicillin-Resistant Staphylococcus aureus Isolated from Clinical Samples at Narayani Samudayik Hospital, Chitwan, Nepal 2021-02-11T19:43:04+00:00 Rama Adhikari Bindu Kshetri Khem Narayan Sharma Santosh Khanal Om Prakash Panta Suprina Sharma Pramod Poudel <p><strong>Objectives: </strong>The main objective of this study was to determine the prevalence of Methicillin Resistance <em>Staphylococcus aureus </em>(MRSA) and MDR bacteria isolated from various clinical specimens from the patients attending Narayani Samudayik Hospital, Chitwan</p> <p><strong>Methods: </strong>A cross sectional study was carried in NPI-Narayani Samudayik Hospital, Chitwan from June to December 2017. Altogether, 3610 clinical specimens mainly pus, blood and urine were collected, streaked on Mannitol Salt Agar and Blood Agar and incubated at 37°C for 24 hours. The confirmed colonies of <em>S. aureus </em>were sub-cultured on Nutrient Agar. The antibiotic susceptibility pattern of all isolates <em>S. aureus </em>was determined by Kirby Bauer disc diffusion method. Isolates resistant to cefoxitin (30mcg) were confirmed as MRSA.</p> <p><strong>Result: </strong>Among 3610 total clinical samples, 17.6 % (635/3610) showed growth and 95(14.96%) <em>S. aureus </em>were isolated. Higher number of <em>S. aureus </em>was isolated from pus sample (93.15%). Out of 95 <em>S. aureus </em>isolates, 55 (57.89%) were identified as MRSA while 40 (42.10%) were MSSA. Vancomycin, ceftriaxone and chloramphenicol were found to be most effective antibiotic against isolates. Whereas, the least effective antibiotic was cefoxitin followed by amoxiclav, oxacillin and amoxicillin.</p> <p><strong>Conclusion: </strong>This study concludes that the overall prevalence of MRSA and MDR among the bacterial isolates is higher compared to other studies. So, it is recommended to monitor the antibiotic susceptibility pattern of pathogens regularly and study the epidemiology of such isolates. &nbsp;</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Methicillin Resistant and Biofi lm Producing Staphylococcus species Isolated from Different Clinical Specimens and Antibiotic Susceptibility Pattern of Isolates 2021-02-11T19:43:05+00:00 Pawana Pandey Anup Bastola Beena Shrestha Puspa Raj Dahal Pradeep Kumar Shah <p><strong>Objectives: </strong>To determine prevalence of Methicillin Resistant <em>Staphylococcus aureus </em>in different clinical specimens and biofilm production along with antimicrobial susceptibility pattern of isolates.</p> <p><strong>Methods: </strong>Cross-sectional study was conducted from September 2019 to February 2020at Sukraraj Tropical and Infectious Disease Hospital. Total 3091 clinical specimens like blood, urine, sputum, pus, swab, body fluid were processed. Identification was done on the basis of colony characteristics, gram staining, culture in Mannitol Salt Agar, coagulase and oxidation fermentation test. Antibiotic susceptibility test and biofilm detection were performed by Kirby Bauer’s disc diffusion methods and Tissue Culture Plate technique (TCP) respectively. Methicillin resistant <em>Staphylococcus </em>species were detected by using Cefoxitin disc.</p> <p><strong>Results: </strong>Out of 52 <em>Staphylococcus </em>species, 39 were <em>Staphylococcus aureus </em>and 13 were Coagulase negative <em>Staphylococcus </em>species. Highest numbers of <em>Staphylococcus </em>species were isolated from blood Sixteen (30.8%) were Methicillin resistant <em>Staphylococcus aureus </em>(MRSA) and 5(9.6%) were Methicillin resistant Coagulase negative <em>Staphylococcus </em>species. There was no significant association (p=0.25) between age group and prevalence of MRSA, MSSA, MRCoNS and MSCoNS. Methicillin resistant <em>Staphylococcus </em>species were resistant to antibiotics like amoxicillin, cloxacillin, erythromycin and higher sensitivity was found in gentamycin. Among 52 Staphylococcal isolates, 11(21.1%) were biofilm producers and 41(78.9%) were non biofilm producers. 90.9%of 90.9% of Biofilm producing <em>Staphylococcus </em>species were resistant towards penicillin and erythromycin</p> <p><strong>Conclusion: </strong>The study shows Methicillin resistant <em>Staphylococcus </em>species were resistant to most antibiotics and rate of resistance was slightly higher in biofilm producing isolates comparing to other isolates. resistance. Regular surveillance of methicillin resistance <em>Staphylococcus </em>species and routine screening of biofilm production is important. &nbsp;</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Effectiveness of Commonly Used Antibiotics in Combination with Honey Against Bacterial Infection 2021-02-11T19:43:07+00:00 Niraj Shrestha Shova Shrestha Bharat Mani Pokharel <p><strong>Objectives</strong>: The study was carried out to compare the inhibitory effects between commonly used antibiotics and bee honey samples, so as to correlate the inhibitory effects between bee honey alone and in combination with antibiotics.</p> <p><strong>Methods</strong>: This study was carried out between December 2012 to September 2013. A total of one hundred and twenty-two clinical microbiological specimens and five different floral sourced honey samples were collected between December 2012 to September 2013. Twenty-three multi-drug resistant organisms were selected. Then, AST for commonly used antibiotics, honey alone and combination of honey-antibiotics discs was done. The difference in ZOI of antibiotic contrasting with the antibiotics containing honey were statistically analysed to define the synergism.</p> <p><strong>Results</strong>: The inhibition due to honey is variable among bacteria types (F=39.17, p&lt;0.05). From means plot, <em>Staphylococcus </em>and <em>Acinetobacter </em>were recognized as highly susceptible bacteria for honey (<em>Χ</em> = 21.1 ± 6.2 mm and <em>Χ</em> = 18.3 ± 3.3 mm respectively) but <em>Acinetobacter </em>species could not show synergism to honey-antibiotic combination. The tested organisms from <em>Enterobacteriaceae </em>family showed effective susceptibility to Chloramphenicol-honey mixture. Imipenem-honey combination and Gentamicin-honey combination showed significant effects against <em>Pseudomonas aeruginosa</em>.</p> <p><strong>Conclusion</strong>: Thus, honey can be used in various bacteria-directed infections and found to be effective in various infections. Incorporation of honey in antibiotics like Chloramphenicol, Imipenem, and Gentamicin work better in healing various infection. &nbsp;</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Incidence of Intestinal Parasites in Government and Private School Going Children 2021-02-11T19:43:08+00:00 Kritika Mishra Freeda Maharjan Reshma Chitrakar Nikita Chhetri Monica Shrestha Richa Chaudhary Shashi Bhushan Chaturwedi <p><strong>Objective: </strong>Aim to assess the incidence of intestinal parasites in government and private school going children.</p> <p><strong>Methods: </strong>The work was conducted from October, 2018 to March, 2019 at Microbiology Laboratory of DAV College, Dhobighat, Lalitpur. A total of 100 stool samples of children aged between 5-12 years were collected from both government and private schools situated in Lalitpur metropolitan city, during school hours. The stool samples were examined for intestinal parasites by Saline wet mount; Iodine wet mount and Formal – ether sedimentation technique. The questionnaires accompanying the queries related to the study were filled.</p> <p><strong>Results: </strong>Of the total 100 stool samples examined, intestinal parasites were observed in 7% (7/100) of the total stool samples. Among the positive stool samples, 71% (5/7) of the stool samples were from government school’s children whereas 29% (2/7) were from private school’s children. Fifty seven percentage 57% (4/7) girls and 43% (3/7) boys were found to be infected with intestinal parasite in the tested stool samples. Out of total parasite detected, 57% (4/7) were eggs of Ancylostoma <em>duodenale</em>, 29% (2/7) were eggs of Ascaris <em>lumbricoides </em>and 14% (1/7) were cysts of Giardia <em>lamblia. </em>The study indicates that <em>Ancylostoma </em>is the most commonly infecting parasite followed by <em>Ascaris </em>and <em>Giardia</em>.</p> <p><strong>Conclusion</strong>: Personal hygiene and sanitary condition were responsible for the incidence of intestinal parasites in the school going children. Environmental sanitation improvement and health education promotion will be helpful to reduce the parasitic infection rate.</p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Detection of Metallo-β-Lactamases and Carbapenemase Production Pseudomonas aeruginosa Isolates from Burn Wound Infection 2021-02-11T19:43:10+00:00 Reshma Maharjan Basudha Shrestha Sanjit Shrestha Khadga Bikram Angbuhang Binod Lekhak Krishus Nepal Milan Kumar Upreti <p><strong>Objective</strong>: The study aims to detect carbapenemase producing <em>P. aeruginosa </em>isolated from burn wounds and confirm MBL production by Imipenem-Combined disk method.</p> <p><strong>Method</strong>: A total of 310 non-repeated clinical specimens including tissues, pus aspirates, and wound swabs were processed using standard microbiological procedure. Each identified isolate of <em>P. aeruginosa </em>was subjected to <em>in vitro </em>antibiotic susceptibility test by using modified Kirby-Bauer disc diffusion method. Two imipenem (10μg) disks were placed on the surface of the agar plate in which one of them was added with 5μl of 0.5M EDTA solution. The result was interpreted after 18 hours of incubation at 37ºC by comparing the inhibition zone of imipenem and imipenem-EDTA disks. The increase in inhibition zone by ≥7mm with imipenem-EDTA disks than imipenem alone was considered as MBL Positive. Similarly, for detecting carbapenemase Modified Carbapenem Inactivation Method (mCIM) was used.</p> <p><strong>Results</strong>: <em>P. aeruginosa </em>was found to be the predominant organism (13.99%). Among 20 <em>P. aeruginosa </em>isolates resistant to imipenem and meropenem, 20% were found to be carbapenemase producer by mCIM assay and 15% were found to be MBL producers by Imipenem-Combined disk method. High percentage of MBL producing isolates of <em>P. aeruginosa </em>were found resistant towards tested antibiotics.</p> <p>&nbsp;<strong>Conclusion</strong>: This study reports that the clinical isolates of <em>Pseudomonas aeruginosa </em>have the ability to produce MBL. The increasing and rapid spread of <em>P</em>. <em>aeruginosa, </em>as well as the rate of drug resistance among the isolates, was found to be a worrisome situation. </p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Phenotypic detection of Extended Spectrum Beta lactamase production from E. coli and K. pneumoniae in urinary samples among children 2021-02-11T19:43:12+00:00 Kalyan Subedi Farisna Karki Sanju Lama Agya Pandey Unita Dahal Rabin Paudyal <p><strong>Objectives</strong>: The main objective of this study was to detect antimicrobial drug resistance (AMR) and Extended Spectrum Betalactamase (ESBL) production phenotypically in <em>E. coli </em>and <em>K. pneumoniae </em>isolated from urines with significant bacteriuria.</p> <p><strong>Methods</strong>: This cross-sectional study was carried out in Microbiology laboratory of Kathmandu College of Science and Technology, Kamalpokhari. The urine samples from suspected urinary tract infected cases were collected from both genders of children below 15 years of age from Out and In-patient department of International Children Friendship Hospital, Maharajgunj and those with significant bacteriuria were cultured for isolating the bacterial etiology targeted as <em>E. coli </em>and <em>K. pneumoniae</em>. AMR for these two bacteria were tested and detected using Kirby Bauer Disc Diffusion technique. ESBL production was confirmed by Double Disc Synergy test (DDST) and Phenotypic Confirmatory Disc Diffusion Test (PCDDT) after screening for all the isolates showing resistance to third generation cephalosporin namely Cefotaxime and Ceftriaxone according to CLSI instructions.</p> <p><strong>Results</strong>: Out of 388 urine samples processed, 29.89% (116/388) showed significant bacterial growth. Five (5) different Aerobic Gram Negative bacterial species were detected and identified. <em>E. coli </em>topped the list (70.68%) followed by <em>K. pneumoniae </em>(15.52%), <em>K. oxytoca </em>(8.62%), <em>Proteus vulgaris </em>(3.45%) and <em>Pseudomonas aeruginosa </em>(1.73%). Among positively screened (44.82%) beta lactamase producers (36.2%) of total isolates were confirmed to produce ESBL. Among ESBL producing isolates, highest susceptibility was seen to Ceftazidime (23.80%) followed by Cefotaxime (16.67%). The ESBL producing isolates were least susceptible to Ceftriaxone (2.38%). AMR was detected using Kirby- Bauer Disc diffusion technique. Comparatively less resistance to amikacin and nitrofurantoin (19.1% and 9.53% respectively) was seen among ESBL producers. 40 out of the 42 (95.23%) ESBL producing strains showed susceptibility to the combination drug, piperacillin/tazobactam. The resistance to meropenem was observed to be less (9.53%) as compared to that to imepenem (7.15%).</p> <p><strong>Conclusion</strong>: This study concluded that there is high prevalence of multidrug resistant uropathogenic clinical strains of <em>E. coli </em>and <em>K. pneumoniae </em>with higher rates of ESBL production. A resistance to the carbepenems is also emerging. Appropriate antimicrobial regimen selection for empirical therapy is thus important for such cases. On managing the empirical antibiotics practice, one can reduce the risk of ESBL producers. There is an essence need of regular routine practice of ESBL detection. </p> 2020-12-26T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Evaluation of Ground Water Quality of Kathmandu Valley and Antibiotic Susceptibility test against Klebsiella pneumoniae 2021-02-11T19:43:13+00:00 Samita Ghartimagar Puja Khatri Swekshya Neupane Dev Raj Joshi Tista Prasai Joshi <p><strong>Objectives: </strong>The aim of this study was to assess quality status of ground water in Kathmandu valley and describe the antibiotic susceptibility of the isolated <em>Klebsiella pneumoniae. </em></p> <p><strong>Methods: </strong>A total of 100 samples were collected from different places of Kathmandu valley with 50 each from two different groundwater sources namely boring and well. This study was conducted from June to September, 2019 at Environment and Climate Study Laboratory, Nepal Academy of Science and Technology (NAST). The physicochemical analysis of the samples was done according to standard methodology. Membrane filtration technique was performed for the enumeration of total coliform and different biochemical tests were performed for isolation and identification of <em>Klebsiella pneumoniae </em>followed by Kirby-Bauer disc diffusion method for antibiotic susceptibility test.</p> <p><strong>Results: </strong>This study reveals the poor microbiological aspects of ground water sources as 98% of total water samples crossed the standard value for total coliform count. The pH, turbidity, ammonia, nitrate and iron content were found to be higher than Nepal Drinking Water Quality Standard (NDWQS 2005) in 15%, 26%, 34%, 7% and 26% of total water samples respectively. The chloride and arsenic content in all the water samples were within the NDWQS, 2005. The 12 out of 18 isolates of <em>Klebsiella pneumoniae </em>from ground water source were highly resistant against first generation Cefazolin however, 15 out of 18 isolates were sensitive to Chloramphenicol. Additionally, four isolates showed zone of inhibition in intermediate range provided by Clinical and Laboratory Standard Institute (CLSI) guideline towards Imipenem and Meropenem.</p> <p><strong>Conclusion: </strong>This study concludes that ground water sources were heavily contaminated by coliform bacteria and most of the physicochemical aspects were under standard limit. Although <em>Klebsiella pneumoniae </em>isolated from ground water were not multidrug resistant, one isolate was recorded to be resistant to Meropenem. These results alarm for circulation of antibiotic resistance in environmental bacterial isolates. Therefore, the appropriate water purification methods should be applied before consumption and should be examined periodically.</p> 2020-12-27T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Characterization of Intestinal Parasitosis in Pregant Women at Ram Janaki Hospital, Janakpurdham 2021-02-11T19:43:15+00:00 Khushbu Yadav Satyam Prakash Basant Kumar Yadav <p><strong>Objectives: </strong>The objective of this study was designed to focus the prevalence, detection and identification of intestinal parasites and its associated factors among pregnant women.</p> <p><strong>Methods: </strong>Total 264 stool samples were collected in a labeled dry, clean disinfectant free wide mouthed plastic container during antenatal visits at Ram Janaki Hospital, Janakpurdham and were examined by macroscopically and microscopically. The detection and identification of protozoal cysts, oocysts, trophozoites and helminthic eggs or larva was done by wet preparation and formalether sedimentation concentration technique. The data was analysed using SPSS 20 version and Microsoft Excel 2007. A Chi-square test was performed to predict the parasite detection using predictor variables. The p-values &lt;0.05 was considered as significant.</p> <p><strong>Results: </strong>The prevalence of intestinal parasitosis among pregnant women was 42%. There was positive association of symptoms of intestinal parasitosis among pregnant women (p &lt; 0.05). The most predominant intestinal parasites among study participants were <em>E. histolytica </em>(20%) slightly dropped by <em>G. lamblia </em>(16%) followed by Hook worm (13%) and <em>A. lumbricoides </em>(11%). The correlation between all the variables with intestinal parasites presence and absence was statistically significant (p&lt;0.05) but statistically insignificant for age and consumption of green leafy vegetables (p&gt;0.05).</p> <p><strong>Conclusion: </strong>The overall prevalence of intestinal parasitosis was relatively moderate. Lack of awareness, low hygienic and sanitation habits regarding parasitic infections were the major determinant factors for higher prevalence. Improving sanitation, awareness creation and public health programes should be organized at regular interval in community. &nbsp;</p> 2020-12-27T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Screening of Indigenous Yeast From Different Ecological Region of Kathmandu Valley and Its Application in Wine Production 2021-02-11T19:43:17+00:00 Bipanab Rajopadhyaya Bipana Maharjan Roshani Maharjan Amrit Acharya <p><strong>Objectives: </strong>The aim of the study was to isolate and screen the potent yeast from the air for implementing new yeast in wine fermentation.</p> <p><strong>Methods: </strong>In this study, 35 air samples collected in sterile grape juice in glass jar and left over for four days exposure for the growth of yeast from different locations around the Kathmandu Valley. Yeasts were screened by culturing on selective Ethanol Sulfite Agar (ESA) media at 30°C for 2-3 days in Microbiology Lab of Pinnacle College. Yeast isolates were characterized based on colony morphology, microscopic characteristics, Fermentative capacity, Hydrogen sulfide production. Selected yeast isolates were subjected to ethanol fermentation and tested for alcohol tolerance capacity. Wine quality was assessed by sensory evaluation.</p> <p><strong>Results: </strong>Of 35 samples, only 20 yeast isolates were isolated. Among these isolates, the variation in colony characteristics along with oval and ellipsoidal microscopic appearance was observed. All the isolates were able to ferment major sugars such as glucose, fructose and sucrose, but few could not ferment galactose and maltose, while none-fermented lactose and xylose. Here, isolates showing no H2S (L29, L34) and mild H2S producer (isolate L31) were subjected to ethanol fermentation. Also, Comparative analysis was made by using commercial standard wine yeast (STAN). Rapid fermentation of grape juice with initial 21 0Brix was observed in L31 isolate, which produced 12.99% v/v alcohol with titratable acidity (TA) 5.25 g/L, followed by L29 strain with 11.99%v/v alcohol and 4.5 g/L TA which were higher than STAN (10.99% alcohol). These isolates specified as Ethanol tolerance up to 13%v/v, while none of them were able to grow at 15% v/v ethanol concentration and 45°C temperature. However, significant growth was observed at pH 3 along with sugar tolerance capacity at 30 0Brix. The wine produced by these isolates was found to be remarkably different among each other. While the sensory analysis of wine led to isolate L31 being congenial to tasters.</p> <p><strong>Conclusion: </strong>L31 isolate was found to be efficient and advantageous for wine production indicating its industrial application. &nbsp;</p> 2020-12-27T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Field Evaluation of Native B. thuringiensis Isolates Against Aphids (Aphis fabae) 2021-02-11T19:43:18+00:00 Ganga GC Charu Arjyal <p><strong>Objectives</strong>: The purpose of this study was to evaluate the aphicidal activity of native <em>Bacillus thuringiensis </em>(Bt) strains.</p> <p><strong>Methods</strong>: Soil samples of Provinces 2 and 3 of Nepal were collected randomly for isolation of Bt by acetate selection method. Bt were identified by observing insecticidal crystal proteins (ICPs) by Commassie Brilliant Blue (CBB) staining technique. Aphicidal activity of 12 <em>B. thuringiensis </em>isolates was evaluated by two processes. The preliminary screening was done by spraying the suspension containing the spore and ICPs mixture in <em>Phaseolus </em>species heavily infested with black aphids (<em>Aphis fabae</em>) in fi elds. The second process (selective bioassay) was done by counting the number of aphids (nymphs, instar, winged, wingless) before and after spraying 5ml of suspension containing the spore and ICPs mixture on the leaf or on the beans pods surface infested by Aphids. The mortality percentage of Aphids after treatment was calculated on the 4th day, by counting the live aphids and the result was recorded.</p> <p><strong>Results: </strong>Preliminary screening for aphicidal activity revealed that 4 isolates ML5(1), CW1(1), SN2(1) and MP2(1) producing spherical crystal protein, showed 100% mortality against nymphs, instar, winged and wingless Aphids. Isolates were effective in controlling the Aphid (<em>Aphis fabae</em>) within 4 days and the part of the plant that was sprayed becomes free of Aphids. Selective bioassay of native isolate MP3(3) was most effective in killing 95.83% of aphids followed by CW2(1), 85.71%, ML5(1), 77.34%, SN3(1), 72.72%, CW1(1), 70.21%.</p> <p><strong>Conclusion: </strong>This study revealed that indigenous <em>Bacillus thuringiensis </em>of Terai region of Nepal are effective in controlling Aphids. &nbsp;</p> 2020-12-27T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Bacteriological Profile and Antibiotic Susceptibility Pattern of Isolates of Wound Infection In Children Visiting Kanti Children Hospital 2021-02-11T19:43:20+00:00 Gita Ghimire Chaudhary RP Binod Lekhak <p><strong>Objectives</strong>: The objectives of this study was to isolate and identify the bacterial etiological agent of wound infection and explore the status of methicillin-resistant <em>Staphylococcus aureus </em>( MRSA), multidrug Resistant (MDR) and extended spectrum β-lactamase (ESBL) producers’ strains in clinical specimens and to find the antibiotic susceptibility pattern.</p> <p><strong>Methods</strong>: A prospective cross sectional study design was conducted from period of February 2014 to October 2014 at Kanti Children Hospital, Kathmandu. The organisms were isolated and identified from pus sample by standard microbiological methods. Antimicrobial susceptibility test was performed by modified the Kirby Bauer disc diffusion method to evaluate the status of MRSA and MDR. ESBL detection was performed by the combined disc diffusion method.</p> <p><strong>Results</strong>: Out of 365 specimens collected between the age group below &lt; 2 to 15 years, 210 (57.73%) samples from male patients and 155 (42.47%) from female patients. In the total samples processed, Gram-positive organisms were found to be more prevalent in which <em>Staphylococcus aureus </em>accounts for 135(47.20%), followed by <em>P</em>. <em>aeruginosa 62 (21.67%), E. coli 29 (10.20%), K. pneumoniae </em>27 (9.44%), <em>Acinetobacter </em>spp. 20 (6.70%), <em>P. vulgaris </em>7 (2.44%) and CoNS 6 (2.10%). Among the <em>S. aureus </em>isolates<em>, </em>29 (21.48%) were found MRSA. Of the total Gram-negative organisms isolated, 74 (51.03%) were MDR and 14 (100%) ESBL producer, (P&lt;0.01). <em>S. aureus </em>was found to be the most important and leading cause of wound infection in this study.</p> <p><strong>Conclusion</strong>: Thus, routine antibiotic susceptibility testing is recommended for empirical drug therapy and proper management of disease. &nbsp;</p> 2020-12-27T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University Characterization of β-Galactosidase from Lactose Utilizing Yeast Isolated from the Dairy Sample 2021-02-11T19:43:21+00:00 Bivek Dahal Sujan Karki Nabaraj Adhikari Upendra Thapa Shrestha <p><strong>Objectives: </strong>The objective of the study was to isolate lactose positive yeasts from dairy samples collected from local markets of Kathmandu, to extract crude β-galactosidase from the lactose positive yeast and to characterize the enzyme for optimum time duration, pH, temperature, Michaelis-Menten constant (K<sub>m</sub>) and maximum activity (V<sub>max</sub>).</p> <p><strong>Methods: </strong>Four lactose positive yeast strains were isolated from dairy samples collected from local market of Kathmandu by pour plate method. Single strain having maximum lactose positive activity was selected for the study. The mass culture of the lactose positive yeast strain was lysed by 2% Chloroform and the yeast cell lysate containing β-galactosidase (i.e. crude enzyme extract) was characterized by using ONPG (Ortho-Nitrophenyl-β-D-galactopyranoside) as substrate. ONPG is a colorless substrate for the enzyme assay which is hydrolyzed by the enzyme into yellow colored product ONP (Ortho-Nitrophenol). The concentration of product formed was monitored spectrophotometrically at 420 nm to determine the enzyme activity and to characterize the enzyme.</p> <p><strong>Results: </strong>The enzyme had wide range of working temperature from 0-50ºC, with optimal temperature of 37ºC. However, greater than 50% hydrolyzing ability was maintained in the range of 14-40ºC. Optimum time of reaction was 70 min. The enzyme had maximum activity in the near neutral pH of 6.8. Michaelis-Menten constant of the enzyme was found to be 2.23 mM of ONPG and Vmax was 58.82 nmol/min/ml. Enzyme activity was 27.88 nmol/min/ml, Specific enzyme activity was 59.97 nmol/ min/mg and total enzyme activity was 3346.33 nmol/min.</p> <p><strong>Conclusion: </strong>The activity over a wide range of temperature 0-50ºC with low K<sub>m</sub> value shows that the enzyme has a commercial application in clearance of lactose pollution in waste water in different environmental conditions.</p> 2020-12-28T00:00:00+00:00 Copyright (c) 2020 Central Department of Microbiology, Tribhuvan University