Nepal Journal of Biotechnology
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Comparative Assessment of Antibiotic Resistance in Lactic Acid Bacteria Isolated from Healthy Human Adult and Infant Feces
Lactic acid bacteria are normal inhabitants of the gastrointestinal tract of humans. Their occurrence in infant and adult feces is abundant. The current study assesses and compares the antibiotic resistance in lactic acid bacteria isolated from healthy human adult and healthy infant fecal samples. A total of 255 lactic acid bacteria isolates (126 from adult feces and 129 from infant feces) were isolated and characterized from 60 fecal samples. Lactobacillus spp., Pediococcus spp. and Enterococcus spp. were included in the study. The study was done using the WHONET software for the analysis of antibiotic susceptibility data of lactic acid bacteria. Most of the Lactobacillus and Pediococcus strains were sensitive to vancomycin. Enterococcus strains showed resistance against vancomycin. Ampicillin, ciprofloxacin and cefuroxime resistance were significantly (p<0.05) higher in Lactobacillus strains isolated from adult fecal samples than those isolated from infant fecal samples. A similar pattern was observed in Enterococcus strains with erythromycin, gentamycin and tobramycin resistance. Pediococcal isolates from adult feces showed significantly higher resistance against tobramycin, ciprofloxacin, gentamycin, cefotaxime and cefuroxime in comparison with infant fecal isolates. Antibiotic resistance was exhibited by lactic acid bacteria against most commonly used antibiotics and it was higher in strains isolated from adult fecal samples than in the strains isolated from infant fecal samples. The increasing trend in antibiotic resistance from infant to adult might be due to food habits and antibiotic intakes. Thus, the widespread antibiotic resistance in different lactic acid bacteriamay pose a food safety concern as well
Nipah Virus (NiV) Infection: Is Nepal Prepared for the Possible Outbreak?
After 20 years of the first Nipah Virus (NiV) outbreak in the world, it re-emerged as the outbreak in India. WHO has recognized NiV as a potent epidemic threat to human health. Both animal-to-human and human-to-human transmission of zoonotic NiV has been documented. Fruit bat of Pteropodidae family is the natural reservoir of the virus. Thus, the territorial habitat of these bats is the high risk zone of NiV outbreak. The symptoms are very nonspecific and the pathogenicity of NiV is yet to be fully understood. Diagnosis of NiV infection still relies on molecular techniques. Till date, no drugs or vaccines against NiV has been approved. Some research have presented arrays of the possible treatment and prevention option, but without sure shot implications. So, appropriate precautions are the only currently available prevention option. Nepal is yet to experience a NiV outbreak but that does not undermine the risk posed to the general population. High risk countries including Nepal should be well prepared to tackle the possible outbreak in future
In Vitro Comparative Study of Antioxidant and Antibacterial Activity of Selected Dietary Plants
Ethanolic extracts of Garlic (Bulb), Aloe (leaf), Flower bud (buds), Turmeric (rhizomes) and Ginger (rhizomes) were used for relative analysis of antioxidant and antimicrobial activity. Antioxidant activity was determined by DPPH [1, 1-Diphenyl-2-picryl hydrazyl] assay and expressed with Ascorbic acid. It was observed that turmeric and ginger have more antioxidant activity than garlic, Aloe and Flower bud. These extracts were further studied for antibacterial activity by agar well diffusion and spectrophotometric method against tetracycline as reference. The result showed that Flower bud is more effective against Escherichia coli, Pseudomonas aeuroginosa, Bacillus subtilis and Staphylococcus aureus compared to other plants extract. However, all the plants extract did show antioxidant and antibacterial activity
Isolation and Screening of Antibiotics Producing Streptomyces spp from the Soil Collected around the Root of Alnus nepalensis from Godawari
Actinomycetes are considered as the most invaluable prokaryotes whose genome mining show a great number of putative secondary metabolite biosynthesis pathways as well as gene clusters ranging from 20 to 50 per genome. The genus Streptomyces has been explored for its ability to produce 60% antibiotics worldwide. Alnus nepalensis (Alder) has been found to harbor diverse Eubacteria in its rhizosphere. To evaluate the antibiotic production potential from Actinomycetes, we collected soil samples from rhizosphere (5-7 cm deep) of Alder tree. Primary screening was done by cross-streak method against Multidrug Resistant (MDR) such as Methicillin resistant Staphylococcus auereus (MRSA), Vancomycin resistant Enterococcus feacalis (VRE), Imepenem resistant Acinetobacterbaumannii, Vancomycin resistant Klebsiella pneumonia and Imepenem resistant E. coli as well as Non-MDRs (E. coli, Bacillus subtilis, Klebsiella pneumoniae, S. aeureu and Enterococcus feacalis). Extraction of antibiotics was done using rota-vapour from extract obtained by solid-substrate fermentation technique followed by solvent extraction. Secondary screening was done using well diffusion assay against MDRs. Among total of 40 isolates of Actinomycetes recovered, 14 showed remarkable zone of inhibition (ZOI) to various MDRs. NASA 303 showed 26 mm of ZOI against VRE, NASA 101 had ZOI of 34 mm against MRSA, NASA 319 had 33.7 mm ZOI against Imepenem resistant E. coli, NASA 306 had 36 mm of ZOI against Vancomycin resistant Klebsiella pneumoniae, and NASA 108 showed ZOI of 29.6 mm against Imepenem resistant E. faecalis. This investigation revealed that the Actinomycetes found in Rhizosphere of Alder tree had MDR killing potent antibiotics, which needs to be further explored
Phenotypic characterization of beta-lactamases producing Gram-negative bacteria in a tertiary hospital, Nepal
Infections caused by beta-lactamases producing Gram-negative bacteria are increasing, thus posing a challenge to the management of such infections. The surveillance data of such bacteria is limited in Nepal so this study aimed to detect the beta-lactamase producing Gram-negative bacteria in a tertiary setting. A total of 604 clinical samples, including urine, blood, sputum and body fluids, were cultured and identified by the routine standard laboratory protocols. Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method following Clinical and Laboratory Standard Institute guidelines (2014). Extended-spectrum beta-lactamases (ESBL) producers were identified by combined disk method and metallo-beta-lactamases (MBL) producers were identified by Imipenem- EDTA combined disk method. Out of 604 samples, 282 (46.7%) samples showed significant growth, of which 229 (81.2%) were Gram-negative bacteria. Of 229 Gram-negative bacteria, 200 (87.3%) were multidrug resistant, 67 (29.3%) were ESBL producers and 16 (7.0%) were MBL producers. Klebsiella pneumoniae were among higher ESBL producers and Pseudomonas aeruginosa were among higher MBL producers. The findings suggest higher antibacterial resistance among Gram-negative bacteria with the added burden of beta-lactamase production. Imipenem was effective against 125 of 229 Gram-negative bacteria tested. Thus, imipenem can be the drug of choice for empirical management. The higher multidrug resistance and higher beta-lactamases production among Gram-negative bacteria warrant the continuous monitoring, surveillance, early detection, and infection control practices of such bacteri
Vancomycin Intermediate MRSA Isolates Obtained from Retail Chicken Meat and Eggs Collected at Pokhara, Nepal
Antimicrobial resistance among food animal isolates is increasing as a result of their uncontrolled uses. The monitoring of antibiotic resistance among these isolates is very necessary. S aureus was isolated from eggshells and chicken meat samples collected from different retail outlets of the Pokhara metropolitan. Samples were inoculated on Mannitol salt agar aseptically and inoculated overnight. Isolated yellow colonies were further examined by Gram-staining, catalase, and coagulase test to detect S aureus. Methicillin resistance was screened using cefoxitin disc. Vancomycin minimum inhibitory concentration (MIC) of Methicillin-resistant S aureus (MRSA) isolates were determined by the agar dilution method following CLSI guidelines. 139 S aureus were isolated from 205 samples. Among them, 89 were from egg samples (out of 125 samples) and 50 from chicken (out of 80 samples). The overall prevalence of MRSA was 12.94%. Antibiotic resistance was significantly higher in MRSA isolates compared to Methicillin sensitive S aureus (MSSA) isolates. The highest rate of resistance was noted for ampicillin, amoxicillin, and erythromycin while the least resistance was noted for gentamicin and amikacin. Vancomycin minimum inhibitory concentration (MIC) range of the MRSA isolates was 0.25-8μg/ml indicating the detection of both vancomycin-intermediate and sensitive isolates from the samples. This is the first study reporting vancomycin-intermediate S aureus (VISA) isolates from Nepal and indicates the increasing drug resistance among animal isolates. Further surveillance studies about the transmission of these pathogens to humans as well as detail molecular analyses are imminent
Detection of Pyuria by Microscopic Urinalysis as a Marker of Pediatric Urinary Tract Infection
Globally, different diagnostic tests of urinary tract infection (UTI) are in clinical practices. A reliable test can increase the efficiency of the healthcare system, especially in a developing country like Nepal, reducing cost and time. Thus, we accessed the possibility of pyuria detected by microscopic urinalysis as a marker of pediatric UTI. The prospective study was conducted fromJuly2014 to January 2015 at Alka hospital, Lalitpur. Microscopic urinalysis of 353clean-catch urine samples was done by the wet mount method, followed by urine culture by a semi-quantitative method. We confirmed 64 (18.1%) UTI cases by culture, the gold standard for UTI diagnosis. Fever was the most common clinical manifestation in UTI cases. The sensitivity, specificity, positive predictive value and negative predictive value of pyuria detected by microscopic urinalysis to identify UTI were 50%, 70.9%, 27.6% and 86.5% respectively. In 318 febrile cases, the sensitivity, specificity, positive predictive value and negative predictive value of pyuria detected by microscopic urinalysis to identify UTI were 73.2%, 72.6%, 28.3% and 94.8% respectively. The findings suggest pyuria detected by microscopic urinalysis as not a worth while marker of pediatric UTI. But it is a trust worthy marker in febrile pediatric cases
Fecal carriage of Extended Spectrum β-Lactamases (ESBL) Producing Escherichia coli and Klebsiella spp. among School Children in Pokhara, Nepal
Extended-spectrum β-lactamases (ESBL) producing microbes in recent years have been a major problem in developing countries like Nepal, with limited treatment options. This study aimed to determine the prevalence of ESBL producing E. coli and Klebsiella spp. in school children in Pokhara, Nepal. The study was conducted from June to October, 2015 at the microbiology laboratory of Manipal Teaching Hospital, Pokhara, Nepal. Antibiotic Susceptibility Test (AST) was done after isolation and identification of bacterial isolates. Then, presence of ESBL enzymes in E. coli and Klebsiella spp. were tested by combination disc diffusion test using cefotaxime and ceftazidime alone and with clavulanic acid. Out of total 309 school children, 211 (68%) bacterial isolates were detected from stool samples. Among them, E. coli and Klebsiella spp. were detected in 97 (46%) and 39 (19%) stool samples respectively. Bacteria isolated from 14 (5%) stool samples were multi-drug resistant (MDR) positive. After applying combined disk method, 88 (29%) isolates were found to be ESBL producer. Emerging prevalence rate of ESBL producing E. coli and Klebsiella spp. are major problem in medical history. Therefore, rapid need of surveillance for effective management of such MDR-strain is required
Infectious sources of Histoplasmosis and molecular techniques for its identification
Histoplasmosis, a fungal infection caused by Histoplasma capsulatum (H. capsulatum), acquired from contaminated soil with droppings of chicken or birds and found to be distributed in many parts of the world. The prevalence of histoplasmosis has not well studied in Nepal. The common symptoms of acute and epidemic histoplasmosis include high fever, cough, and asthenia and weight loss. Most of the infections associated with histoplasmosis are asymptomatic. People with compromised immune systems such as HIV/AIDS (PLWHA), cancer, and organ transplant recipients are at risk of developing this disease. In this review, we have summarised the current status of histoplasmosis in Nepal and molecular techniques available for its identification. To date, the significant outbreak is not reported in Nepal, but the risk of infection for the vulnerable population cannot be undermined. Appropriate preventive measures and treatment on time can reduce the burden of this fungal disease. Further, this review is also focused on molecular identification of H. capsulatum. Hence, careful considerations by concerned stakeholders for national surveillance programs and the treatment of patients on time after proper diagnosis is highly recommended
Comparative Antioxidant, Antimicrobial and Phytochemical Assesments of Leaves of Desmostachya bipinnata L. Stapf, Hordeum vulgare L. and Drepanostachyum falcatum (Nees) Keng f.
Nepal is rich in varieties of religious plants. The locally used religious plants also carry medicinal importance. Desmostachya bipinnata L. Stapf, Hordeum vulgare L. and Drepanostachyum falcatum (Nees) Keng f. are three plants belonging to the family Poaceae having religious significance in different practices of Hinduism. They were also used as traditional medicines by our ancestors but nowadays they are underutilized. In this research, our core objective was to validate the traditional assumption of use of these plants in medicinal purposes by carrying out the assessments like antimicrobial assessment, antioxidative assessment and phytochemical assessment. Methanolic extracts produced from leaves of all three plants were examined for antimicrobial activities through agar well diffusion method. The same extracts were also assessed for determining their antioxidative potentials with the use of DPPH (1, 1-diphenyl-2-picryl hydrazyl) free radical scavenging assay followed by qualitative phytochemical analysis and GCMS (Gas Chromatography Mass Spectroscopy). Most promising antimicrobial activity was shown by Desmostachya bipinnata L. against Salmonella typhimurium and Staphylococcus aureus, Drepanostachyum falcatum (Nees) Keng f. against Salmonella typhimurium and Klebsiella pneumoniae and Hordeum vulgare L. against Salmonella typhmurium and Staphylococcus aureus. The antioxidant activity of the plant extracts were observed in descending order of Hordeum vulgare L.>Desmotachya bipinnata L. > Drepanostachyum falcatum (Nees) keng f. and phytochemical assessment of the extracts indicated the presence of alkaloids, glycosides, sterols, Triterpenes, Saponins, Flavonoids, Coumarins, Phlobatanin and reducing sugars. Through this project, we can clarify that the above mentioned plants have bioactive compounds which contributed for the presence of antimicrobial and antioxidative property in the plants