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Item BACTERIOLOGICAL ANALYSIS OF MILK SOLD IN KATHMANDU AND ANTIBIOTIC SUSCEPTIBILITY PATTERN OF Staphylococcus species ISOLATED FROM MILK(Amrit Campus, 2022-08-08) Soniya, BohoraMilk is a rich source of nutrients. Milk –borne pathogenic bacteria pose a serious threat to human health. Staphylococcus aureus, Salmonella spp., Listeria monocytogenes with Escherichia coli and Campylobacter are the main microbial hazards associated with contaminated milk. Therefore, it can cause milk borne diseases like scarlet fever, Brucella, diphtheria typhoid etc. This study was conducted to assess and compare microbial quality of raw milk and pasteurized milk and also determine the antimicrobial susceptibility of Staphylococcus species isolated from milk sample consumed in Kathmandu. For this, 30 milk samples (15 raw milk and 15 pasteurized) were collected from different location of Kathmandu district. Total Plate Count and Total Staphylococcus Count for each sample were determined by pour plate technique. While for isolation of Staphylococcus species, samples were isolated by using selective media (MSA) and characterized by biochemical test. Antibiotic susceptibility testing of isolates was carried out by Kirby Bauer disk diffusion method. Total bacterial count of all raw milk samples were within the range while for TBC of pasteurized milk 93% were within the range. 17 Staphylococcus species were isolate from TSC. Among them 15 were identified as Staphylococcus aureus. 17 Staphylococcus species were 100% sensitive to Cotrimoxazole, Amikacin and Levofloxacin but resistant to Penicillin G (100%),Ceftriaxone (52.92%), Tetracycline (17.68%), Cefoxitin (23.58%), Ampicillin (76.82%) Ciprofloxacin (17.68%) and Chloramphenicol (11.79%). 3(17.68%) of Staphylococcus aureus isolated from raw milk samples showed multi-drug resistance and 4(23.58%) MRSA were detected. It is concluded that the milk produced by small scale farm from different places of Kathmandu district are not of quality and can be potential source of milk-borne infection. It is recommended that routine assessment of microbial quality of milk should be done for the safeguard of consumer health.Item EFFICACY OF HAND SANITIZERS AGAINST STANDARD BACTERIAL CULTURES(Amrit Campus, 2022-08-08) Sherpa, Shyronह्याण्ड स्यानिटाइजरहरु हातको स्वच्छता कायम राख्न प्रयोग गरिन्छ । कोरोना महामारीले यसको प्रयोग बारे ध्यान खिचेको थियो । धेरै कम्पनीहरुले उत्पादन गरिरहेका छन् तर प्रभावकारीता राम्रोसँग स्थापित हुन सकेको छैन । यस अध्ययनको मुख्य उद्देश्य काठमाण्डौमा बिक्री हुने ह्याण्ड स्यानिटाइजरहरुको व्याक्टेरियालाई मार्ने क्षमताको प्रभावकारीता मूल्याङ्कन गर्नु थियो । यस अध्ययन मार्च देखि अप्रिल २०२२ सम्म गरिएको थियो । यस अध्ययनमा काठमाण्डौका बजारबाट ३१ वटा अल्कोहल आधारित ह्याण्ड स्यानिटाइजर किनिएको थियो । Hand sanitizers are used to maintain hand hygiene. Corona pandemic had gain the attention; many companies are producing it and are marketed but their efficacy have not been well established. Thus, the main aim of this study was to evaluate the antibacterial efficacy of the hand sanitizers sold in Kathmandu. This study was carried out from March to April, 2022. In this study, 31 alcohol based hand sanitizers were purchased from the markets of Kathmandu. Among them, 15 liquid based hand sanitizers (9 contain alcohol and 6 contain alcohol with additional ayurvedic ingredients) and among 16 gel based hand sanitizers, (10 contain alcohol and 6 contain alcohol and additional ayurvedic ingredients). Efficacy of hand sanitizers were evaluated using standard ATCC cultures: Escherichia coli 35218, Escherichia coli 25922, Staphyloccoccus aureus 43300 and Klebsiella pneumoniae 700603 by agar well diffusion method. In the volume of 50 μl hand sanitizers, (46.67%, 60%, 26.67% and 33.33%) liquid based, (20%, 40%, 33.33% and 26.67%) liquid with ayurvedic ingredients based, (12.5%, 18.75%, 18.75% and 12.5%) gel based and (12.5%, 12.5%, 12.5% and 6.25%) gel with ayurvedic ingredients based hand sanitizers showed the antibacterial effect against E. coli 35218, E. coli 25922, S. aureus 43300 and K. pneumoniae 700603 respectively. In the volume of 100 µl hand sanitizers, (53.33%, 60%, 46.67% and 46.67%) liquid based, (40%, 40%, 40% and 40%) liquid with ayurvedic ingredients based, (50%, 62.5%, 62.5% and 62.5%) gel based and (25%, 31.25%, 25% and 25%) gel with ayurvedic ingredients based hand sanitizers showed the antibacterial effect against E. coli 35218, E. coli 25922, S. aureus 43300 and K. pneumoniae 700603 respectively. In the volume of 150 µl hand sanitizers, (60%, 60%, 60% and 60%) liquid based, (40%, 40%, 40% and 40%) liquid with ayurvedic ingredients based, (62.5%, 62.5%, 62.5% and 62.5%) gel based and (31.25%, 31.25%, 31.25% and 31.25%) gel with ayurvedic ingredients based hand sanitizers showed the antibacterial effect against E. coli 35218, E. coli 25922, S. aureus 43300 and K. pneumoniae 700603 respectively. Comparatively, liquid with ayurvedic ingredients based sanitizers were more effective than gel with ayurvedic ingredients based sanitizers.Item ENUMERATION AND DETECTION OF ANTIBIOTIC SUSCEPTIBILITY PATTERN OF COLIFORM BACTERIA FROM MILK SAMPLES IN KATHMANDU.(Amrit Campus, 2022-08-08) SURAJ, CHAULAGAINThe presence coliforms and their resistance in milk is the big issue in present time. Milk is an excellent source of nutrients and also serves as a good medium for the growth of milk-borne pathogens. Cross-sectional study was conducted to assess and compare microbial quality of raw milk and pasteurized milk and also determine antimicrobial susceptibility patterns of coliforms from the milk samples. For this, 30 milk samples (15 raw and 15 pasteurized milk) were collected from different locations of Kathmandu district. Starch adulteration test and MBRT were done. TCC, FCC for each sample were determined by pour plate technique and interpretated with BIS guidelines (1992), DFTQC guidelines and identification was done. Antibiotic susceptibility testing of isolates was carried out by Kirby Bauer disk diffusion method using 12 different antibiotics. TCC of the 12 raw samples were higher than the guideline and its FCC was also found to be higher in 9 samples. In case of pasteurized samples, TCC was higher in 6 samples and FCC in 4 samples. A total of 31 isolates, 21 from raw samples and rest from pasteurized samples were identified. Out of 31 isolates, 17 (54.84%) were identified as Klebsiella spp., 13 (41.94%) were E. coli and 1 (3.22%) was Citrobacter spp. AST of coliform isolates were 100% sensitive against TE. 96.77% of the isolates were sensitive towards NIT, PIT, COT, C and AK. Out of total, 11 (35.48%) were MDR (Multi-Drug Resistant). Among them, 7 (63.64%) were from raw samples and rest 4 (36.36%) from pasteurized samples. Although, quality of most of the samples were good as per MBRT but the presence of antimicrobial resistant bacteria and adulterants questions the overall quality ofmilk. Thus, it is concluded that the milk produced by small-scale farm from the studied area are not of good quality, caused by coliforms especially the antibiotic resistant. Therefore, such type of study for monitoring the microbial quality of milk should be done in order to safeguard the consumers. Otherwise, it will be hazardous for the consumers and can be a potential source of milk-borne infections.Item SCREENING OF ANTIBIOTIC PRODUCING ACTINOMYCETES FROM THE SOIL SAMPLES AND ANTIBACTERIAL ACTIVITY IN VARYING SODIUM NITRATE CONCENTRATION(Amrit Campus, 2022-08-08) DANGOL, SWORNIMAActinomycetes are gram positive filamentous, slow growing bacteria, best known to produce antibiotic. The aim of this study was to screen antibiotic producing actinomycetes and determine its antibiotic activity against ATCC cultures at different gradient of NaNO3. This study was carried out from March 29 to April 28, 2022. Thirty collected samples were collected and transported and processed in Amrit campus. Primary and secondary screening were performed by perpendicular streaking and agar well diffusion method respectively. Characterization and identification of the isolated actinomycetes were performed. From thirty collected samples, twenty-eight samples were actinomycetes. Only two samples were antibiotic producing actinomycetes MI410 -3 and NP110 -2 showed antibacterial activity against ATCC cultures viz: S. aureus 43300, E. coli 35218, E. coli 25922 and Klebsiella 700603 in primary screening. Antibiotic was produced by sub-merged state fermentation with varying concentration of NaNO3 and secondary screening was done by agar well diffusion against ATCC cultures in comparison to standard streptomycin (100 µg/mL). In comparison to standard streptomycin (100 µg/mL) extract MI410-3 with 1% NaNO3 was effective only against E. coli 25922 while NP110-2 with 0.5% NaNO3 against E.coli 35218 (14.67mm), E.coli 25922 (19.67mm) and S. aureus 43300 (17mm) whereas NP110-2 has also shown antibacterial effect with 1.5% NaNO3 against E. coli 35218 (13mm), Klebsiella spp. ATCC 700603 (11.5mm) and S. aureus 43300 (12.5mm). Statistically, there is significant difference at 5% level of significance between the sample concentration at 0.5% of NaNO3 (.001, P<0.05) and 1.5% of NaNO3 (0.024, P<0.05) in antimicrobial activity. However, there is no significant difference at 5% level of significance between the sample concentration at 1% (0.356, P>0.05). The isolated actinomycetes was presumed as Streptomyces spp. Both MI410-3 and NP110-2 (7.14%) showed antibacterial activity against cultures in primary screening. The indigenous species of actinomycetes, isolated from various places of Kathmandu valley can be used in industrial production of antibiotics which can help in economic growth of Nepal.Item SCREENING OF POTENTIAL ANTIBIOTIC PRODUCING ACTINOMYCETES EXTRACTED FROM SOIL AND WATER SAMPLES FROM DIFFERENT REGIONS OF KATHMANDU VALLEY(Amrit Campus, 2022-08-08) Sardar, Chandra KishoreActinomycetes are Gram-positive, aerobic spore forming bacteria that are characterized by aerial and mycelial growth and are chief antibiotic producers. The aim of this study was to screen antibiotic producing actinomycetes and determine its antibiotic activity against ATCC cultures. A total of 60 samples (30 water and 30 soil) were collected from different regions of Kathmandu Valley viz. Kathmandu, Lalitpur and Bhaktapur spanning from the month of March 2022 to May 2022. Spread plate technique was employed to isolate the actinomycetes on Starch M-Protein Agar, and primary and secondary screening techniques were performed via Perpendicular Streak method and Agar-well diffusion respectively for screening their ability to produce antibiotics. The actinomycetes were confirmed by the macro and microscopic examination, biochemical and physiological tests. The crude extract obtained from the submerged state fermentation was filtered and centrifuged; tested against the Standard cultures viz; Staphylococcus aureus ATCC 43300, Escherichia coli ATCC 35218, Escherichia coli ATCC 25922 and Klebsiella spp. ATCC 700603 via agar well diffusion method. Out of the 28 (93.3%) isolates obtained from 30 soil samples, only two isolates (7.3%) i.e., NP1 and MI4 showed antimicrobial activity against the ATCC cultures which were presumed to be Streptomyces. No actinomycetes were obtained from water samples. ANOVA revealed no significant difference at 5% level of significance (0.535; P>0.05) between the standard streptomycin (100 µg/ml) and NP1.The soil of Kathmandu Valley harbors microbial diversity that encompasses potential antimicrobial producing actinomycetes which in turn can help in booming the economy by enabling the production of indigenous antibiotics.