Browsing by Author "Ghimire, Prakash"

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    Efficacy of Native Bacillus thuringiensis against Mosquito Vector
    (Nepal Health Research Council, 2023) GC , Ganga; Parajuli, Kshama; Gautam, Ishan; Banjara, Megha Raj; Ghimire, Prakash; Rijal, Komal Raj
    Abstract Background: Larval source management is an effective measure to control mosquito-borne diseases. Bacillus thuringiensis produces specific insecticidal crystal proteins toxic to mosquito larvae. In many parts of the South East Asian region, Bacillus thuringiensis is used for larval source management. In Nepal, larvicidal Bacillus thuringiensis is not available. The study aims to isolate larvicidal Bacillus thuringiensis from soil samples of Nepal to control mosquitoes. Methods: Native Bacillus thuringiensis was obtained from soil samples by the acetate selection method. It was identified by observing crystal protein with Coomassie Brilliant Blue stain in a light microscope. The mosquito larvae were collected from different breeding habitats. A preliminary bioassay was performed by inoculating three loopful of 48 hours culture of spherical crystal protein producing Bacillus thuringiensis in a plastic cup containing 25 larvae and 100 ml of sterile distilled water. The cup was incubated at room temperature for 24 hours to observe the mortality of larvae. Further selective bioassay was performed with the isolate which showed 100% mortality, as described above in four replicates along with the negative and positive control. Results: Out of 1385 Bacillus thuringiensis obtained from 454 soil samples, 766 (55.30%) were spherical crystal protein producers, among them, a single strain (14P2A) showed 100% mortality against mosquito larvae. The lethal concentration doses required to kill 50% and 90% of the larval population were 32.35 and 46.77 Parts per million respectively. Conclusions: The native Bacillus thuringiensis produces the crystal protein effective in killing mosquito larvae. The native Bacillus thuringiensis should be included as a tool to control mosquito-borne diseases in Nepal. Keywords: Bacillus thuringiensis; mosquito; Nepal; tool.
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    Epidemiological Characteristics of Hepatitis C Patients Attending a Tertiary Care Hospital
    (Nepal Health Research Council, 2024) Kattel, Hari Prasad; Sharma, Sangita; Alfsnes, Kristian; Pathak, Rahul; Rijal, Komal Raj; Ghimire, Prakash; Andreassen, Ashild K; Banjara, Megha Raj
    Background: Hepatitis C infection is a major public health concern in Nepal. Epidemiological information on hepatitis C virus along with the status of co-infection with hepatitis B virus and human immunodeficiency virus is essential to controlling the hepatitis C burden. The objective of this study was to determine the sero-prevalence of hepatitis C virus infections, proportions of co-infections with human immunodeficiency virus and hepatitis B virus, and identify the demographic characters, and routes of transmission. Methods: A cross-sectional study was conducted from December 2019 to February 2024 at Tribhuvan University Teaching Hospital. The serological tests were performed by enzyme-linked immunosorbent assays from 25133 patients’ serum in four years. Results: The sero-prevalence of hepatitis C virus -infected patients was 0.8% (211/25133). Among them, 6.6% (14/211) were co-infected with human immunodeficiency virus and 1.4% (3/211) with hepatitis B virus. Among 211 hepatitis C virus patients, 174 (82.5%) were male, 156 (73.9%) were young aged 15–47 years with various professions, 167 (79.1%) were literate, and almost one-third of the patients (33.2%, 70/211) were regular alcoholics. Needle sharing among intravenous drug users (45.5%, 96/211) and sexual intercourse (28%, 59/211) were the most common modes of transmission. Conclusions: Although the prevalence of hepatitis C infections is less than 1%, it is more common among young male intravenous drug users. Awareness of the spread of hepatitis C infections among this population needs to be emphasized to control hepatitis C in Nepal. Keywords: characters; co-infection; hepatitis C, sero-prevalence.
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    Extended Spectrum Beta Lactamase Escherichia Coli in Bagmati River, Kathmandu Valley
    (Nepal Health Research Council, 2023) Ghimire, Bindu; Pokherel, Muna Kumari; Banjara, Megha Raj; Rijal, Komal Raj; Ghimire, Prakash
    Abstract Background: Antimicrobial resistance organisms in the peripheral communities of an environment can be predicted by the presence of extended-spectrum beta-lactamase Escherichia coli in that environment. The close connectivity between humans and water sources can facilitate the entry of antimicrobial resistant organisms into the human ecosystem. The aim of this study was to assess beta lactamase producing Escherichia coli from Bagmati river within Kathmandu valley. Methods: In the year 2020, a cross-sectional study was conducted on water samples collected from 66 locations along the Bagmati River. Coliforms were isolated by five tubes dilution method and identified by cultural and biochemical tests. Further Escherichia coli was isolated in eosin methylene blue agar at 44.5 ?C. Antibiotic susceptibility test was performed by Kirby Bauer disk diffusion methods. Beta lactamase gene types were detected by using conventional multiplex polymerase chain reaction. Results: A total of 615 bacterial isolates were identified among which 39 % (n=241) were Escherichia coli. Extended spectrum beta lactamase producing Escherichia coli was confirmed in 16.6 % (40/241) of total Escherichia coli isolates. Among 66 sites this isolate was detected in 26 (40 %) sampling sites excluding upstream regions. All the Escherichia coli isolates were multidrug resistance showing higher percentage (>99 %) of resistant for penicillin, tetracycline and erythromycin antibiotics. There were significant differences in resistance rate for cefotaxime and ceftazidime by extended spectrum beta lactamase producing and non-producing Escherichia coli (p<0.05). Conclusions: Presence of multidrug resistance extended spectrum beta lactamase producing Escherichia coli in river streams suggests the chances of circulating within river system and hence transmitting in human community. Key words: Bagmati river; drug resistance; escherichia coli; human.
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    Metallo-β-lactamase-producing Pseudomonas aeruginosa Isolates from Two Tertiary Care Centres in a District of Nepal: A Descriptive Cross-sectional Study
    (Nepal Medical Association, 2024) Shrestha, Pushpa Man; Kattel, Hari Prasad; Sharma, Sangita; Bista, Pratibha; Basnet, Bhupendra Kumar; Ghimire, Prakash; Rijal, Komal Raj
    Abstract Introduction: Pseudomonas aeruginosa isolates producing metallo-β-lactamase have caused nosocomial outbreaks, severe infections, and ineffective carbapenem therapy worldwide since 1991. Due to their prevalence, hospital infection control techniques are difficult. This study aimed to find out the prevalence of metallo-β-lactamase among P. aeruginosa isolates from two tertiary care hospitals in Kathmandu. Methods: A descriptive cross-sectional study was conducted at the Department of Microbiology and Department of Pathology of two tertiary care centres in Kathmandu from 7 December 2021 to 6 April 2023, after receiving ethical approval from the Ethical Review Board. Isolated strains were identified and tested for antibiotic susceptibility by modified Kirby-Bauer Methods. Metallo-β-lactamase presence was confirmed using an imipenem-imipenem/ ethylenediaminetetraacetic acid disc. A convenience sampling method was used. The point estimate was calculated at 95% Confidence Interval. Results: Among 255, Pseudomanas aeruginosa isolates, the distribution of metallo-β-lactamase-producing Pseudomanas aeruginosa was 103 (40.39%) (34.32-46.69 at 95% Confidence Interval). Multidrug resistance categories included multidrug resistance 74 (71.80%), extensively drug resistance 32 (31.10%), P. aeruginosa difficult-to-treat 16 (15.53%) and carbapenem-resistant P. aeruginosa was determined to be 82 (79.60%). Conclusions: The study found a high prevalence of metallo-β-lactamase-producing Pseudomanas aeruginosa isolates, requiring early identification, infection control measures, and an all-inclusive antimicrobial therapy protocol to reduce their spread in medical settings.
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    Multidrug resistant Escherichia coli isolated at National Public Health Laboratory, Nepal
    (2025) Acharya, Jyoti; Shrestha, Anjana; Rijal, Nisha; Jha, Runa; Rijal, Komal Raj; Sharma, Supriya; Banjara, Megha Raj; Ghimire, Prakash
    Background: Antimicrobial resistance in Escherichia coli is mostly associated with ß-lactamases and carbapenemases enzyme production resulting in treatment challenges. This study was conducted with the aim to detect and characterize antimicrobial resistance in E. coli isolates. Methods: A cross-sectional study was conducted during 2018-2022, at National Public Health Laboratory where the clinical specimens (24636) received were processed for identification and characterization of antimicrobial resistance following conventional & advanced methods. Antibiotic susceptibility tests were performed by Modified Kirby Bauer disc diffusion and Minimum inhibitory concentrations using VITEK2 compact (Biomeriux). The isolates were tested for extended-spectrum ?-lactamases and Carbapenemase production following Clinical Laboratory Standards Institute guidelines. Results: Bacterial growth was observed in 9% (2166/24636) of the specimens, of which 44% (959) were E. coli. Among the 959 E. coli isolates, 320 were reconfirmed with VITEK-MS (Biomeriux). Phenotypic multi-drug resistance was observed in 75% (240/320) of the isolates with 62% (197/320) extended-spectrum ?-lactamases, 12% (39/320) AmpC-ß-lactamase, 10% (31/320) serine carbapenemases and 7% (22/320) Metallo-ß-lactamase while 3% (9/320) produced three types of enzymes. The extended-spectrum-?-lactamase producing E. coli were sensitive to Tigecycline (100%), Amikacin (92%), Imipenem (87%), and Meropenem (84%). Carbapenemase producers were sensitive to Tigecycline (100%), with 61% to Amikacin. Extensive-drug resistance was observed in 2% (7/320) of the isolates, with Colistin resistance in one. Conclusions: The findings highlight alarmingly high antimicrobial resistance in E. coli posing significant challenges in treatment. Early detection of multi-drug resistant isolates in healthcare settings is crucial to combat antimicrobial resistance. Keywords: ?-lactamase; carbapenemases; extensive-drug resistance; metallo-ß-lactamase; multidrug resistance

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