Nepal Journal of Biotechnology
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Cyclospora cayetanensis: An Infestation among Diarrheal Children in Kathmandu Valley, Nepal
Cyclospora cayetanensis, a coccidian parasites which is responsible for recurring diarrhea and gastroenteritis especially among children living under poor hygienic condition in developing country like Nepal. Aimed of this research is to find out the prevalence of intestinal parasites and C. cayetanensis among diarrheal children in a Pediatric Hospital in Kathmandu valley, Nepal. Cross-sectional type of study was done. Altogether 196 stool specimens were collected from June to September 2013 among outpatient diarrheal children in Kanti Children’s Hospital. Modified Ziehl Neelson staining method was applied for detection of oocysts of Cyclospora after formal-ether sedimentation. Parasites were detected in 13.7% (27/196) of stool samples from ≤ 15 year old diarrheal patients. C. cayetanensis was detected in 4.8% (8/196). In genderwise, infection rate of C. cayetanensis 4.5 % (5/112), in male were higher than 3.6% (3/84) in female. In agewise, infection rate of C. cayetanensis in 5.7 % (3/52) 11-15 year old were highest followed by 4.1% (3/78) in 0-5 year old and 3.0% (2/66) in 6-10 year old. In seasonwise, infection rate of Cyclospora was highest in the month of August 7.4% (4/54) followed by 3.9% (2/51) in July, 2.3% (1/44) September and 2.1% (1/47) in June. Altogether five different type of parasites were detected. Infection rate of Giardia lamblia were highest 5.1 % (10/196) whereas lowest was Cryptosporidium parvum 1.0 % (2/196). Prevalence of C. cayetanensis is highly probable to infant, neonate, toddler and diarrheal children. Therefore, attention should be made in laboratory investigation of C. cayetanensis while suspecting the diarrheal patients infected with other parasites
Antimicrobial Resistance Patterns and Plasmid Profiles of Methicillin Resistant Staphylococcus aureus Isolated from Clinical Samples
Methicillin-resistant Staphylococcus aureus (MRSA), showing resistance to several antibiotics is a global health problem associated with considerable mortality and morbidity. Antibiotic susceptibility test is a commonly used method to characterize MRSA in epidemiologic studies. Additionally, plasmid profile has been reported to be useful in tracing the epidemiology of antibiotic resistance. This research was conducted to determine the antimicrobial resistance patterns and plasmid profiles of MRSA isolated from clinical samples at KIST Medical College, Imadol, Kathmandu, Nepal. All the clinical specimens sent to the laboratory were processed by standard microbiological techniques and antibiotic susceptibility testing was done by the modified Kirby Bauer disc diffusion method. Further, plasmid profiling was done by Alkaline-lysis method. A total of 27 (38.02%) MRSA were isolated from 71 S. aureus positive samples. MRSA showed the highest resistance towards penicillin (92.60%) and ampicillin (92.60%). In contrast, high levels of sensitivity were shown towards vancomycin (85.19%) and tetracycline (85.19%). Out of 27 MRSA positive samples, single plasmids were isolated from only 6 (22.22%) MRSA isolates. Antibiograms alone are inadequate to accomplish the characterization of MRSA during epidemiological studies. However, plasmid profile analysis in conjunction with the antibiotic susceptibility pattern is valuable in the epidemiological investigation of MRSA, and for reducing MRSA prevalence and treatment cost
Ethanol Extract of Choerospondias axillaris Fruit Pulp Enhances Haematological Parameters in Oncorhynchus mykiss Cultured in Nepal
The present outdoor experiment was carried out to evaluate the effect of dietary supplementation of lapsi Choerospondias axillaris (Roxburgh, 1832) on haematological parameters in rainbow trout Oncorhynchus mykiss in Nepal. The lapsi fruits were obtained from local market of Kathmandu. The feeding trail was conducted for 90 days. About 270 trout with similar body weight (5±1g) were distributed randomly at the rate of 15 fishes per cage (1m3) into 18 cages placed in raceway pond. Six practical diets containing 40% protein were prepared as T1 (0.0 g kg-1) T2 (0.1 g kg-1), T3 (0.2 g kg-1), T4 (0.4 g kg-1), T5 (0.8 g kg-1) and T6 (1.6 g kg-1) supplemented by the ethanol extract of lapsi fruits along with other usual ingredients viz. fish meal, wheat flour and cod liver oil etc. At the end of the experiment the haematological parameters were measured. A significant difference (P< 0.05) in haematological parameters was observed between the treated diets fed groups to that of control diet fed group. Total erythrocytes count (RBC), Haematocrit (Hct), Haemoglobin concentration (Hb), and erythrocyte indexes (MCV, MCH and MCHC) were found significantly higher in T4 (0.4 %) diet fed trout as compared to the control. A minimum of 0.4 % (0.4g kg-1) lapsi fruit extract in fish feeds gave more increase in haematological parameters of Oncorhynchus mykiss. Inclusion of lapsi fruit extract at 0.4 % concentration is therefore could be used effectively in aquaculture
Isolation, Identification and Production of Encapsulated Bradyrhizobium japonicum and Study on their Viability
Rhizobium, a nitrogen-fixing bacteria is the essential feature of leguminous plants which is essential for the regeneration of nutrient-deficient soil. This study was aimed to isolate, identify, mass culture and immobilize Bradyrhizoium japonicum in encapsulated form and test their viability. Root nodules were sterilized, grinded and cultured aseptically in YEMA media containing Congo red. The obtained colon was sub-cultured to get a pure culture and different biochemical tests were conducted which proved Bradyrhizobium japonicum as the slow-growing species. The test shows a positive result of catalase production and nodulation test whereas the pH tolerance test shows more tolerance to the acidic pH. Similarly, Bradyrhizaobium japonicum can tolerate 1% and 2% NaCl concentration and it doesn’t show resistance to the penicillin disc of 10mg. The mass culture and encapsulation with sodium alginate adding sucrose as nutrient proved the simplicity for handling. Altogether 548 beads were prepared from the 100ml of the cultured broths which were viable for more than 190 days at 1%, 2% and 3% sucrose concentration but less viable at 5% and 10% sucrose concentration under room temperature
Technologies for the Extraction, Separation and Purification of polyphenols – A Review
Polyphenols are high molecular weight, organic molecules mainly found in plant kingdom. They are mostly known for their positive impact on health, specifically for their antioxidant activity. Indeed, they are widely studied for the prevention of multiple diseases such as cancer, inflammatory, cardiovascular and neurodegenerative diseases. Nevertheless, extractions of these growing interest molecules remain challenging using conventional methods such as solvent extraction. That is why recent researches have focused on improving the extraction of polyphenol by using different technologies such as ultrasound, microwave, pressurized liquid, pulsed electric field, supercritical fluid and high hydrostatic pressure. In the current context, the assisted-extraction should demonstrate their potential to improve the extraction efficiency while being cost-effective and with a low environmental impact. To this end, technologies ought to, for instance, increase the solubility of polyphenol and the permeability of the cell wall. Consequently, this review is focused on the use and potential of these technologies to improve polyphenol extractions from plants as well as their purification using various methods. It discusses of the advantages and disadvantages with some examples of all these technologies assisted-extraction in comparison with conventional extraction method as well as purification technology
Nematode Fauna Associated with Kiwi (Actinidia delicosa, Chev.) Plants in Machchhegaun, Kathmandu, Nepal
Plants harbor many trophic groups of nematodes in them. The plant production is also determined by the occurrence of nematodes adjacent to the rhizosphere of plants, such as parasitic, free-living etc. Altogether 40 samples from the 30 cm away from the Kiwi plants were examined to detect the nematode distribution in Kiwi plants in Machchhegaun, Kathmandu, Nepal. Total 10 genera of nematodes including both free living/beneficial and parasitic were identified. Overall, 880 individuals of 10 nematodes genera belonging to four orders were found to be linked with Kiwi plants, among them the highest report was of order Dorylaimida (40.91%) (Dorylaimus spp., Cephalobus spp., Eucephalobus spp. and Discolaimus spp.) which was followed by Mononchida (36.36%) (Iotonchus spp., Parahadronchus spp. and Mononchus spp.), Rhabditida (18.18%) (Rhabditis spp. and Mesorhabditis spp.) and Tylenchida (4.55%) (Helicotylenchus spp.). No any published data about study of plant nematodes was found from study area. So, these four order of plant nematodes have been reported for the first time associated with Kiwi plants from Machchhegaun in Nepal. The result specified no significant distribution of nematodes in all Kiwi plants. Proper management of manures and Kiwi plants treatment is recommended for more production of Kiwi fruits in study area by nitrogen reduction, phosphorus reduction, odor reduction, energy recovery and adding value to manure techniques
Evaluation of Phytochemical, Antimicrobial, Antioxidant Activity and Cytotoxic Potentials of Agave americana
Ethnomedicinal plants are being used as a source of medicine from ancient time but they lack the proof of modern scientific evidence for their effectiveness. This study focuses on the evaluation of phytochemical, antimicrobial, antioxidant properties of one of the ethnomedicinal plant Agave americana from Dhulikhel region of Nepal. The plant extract was prepared using solvent-based warm soxhlet extraction from the leaves of the plant and antimicrobial activity against six different non-resistant clinical isolates of bacteria (Staphylococcus aureus, Shigella, Klebsiella pneumoniae, Escherichia coli, Bacillus thuringiensis, and Salmonella paratyphi) was evaluated using agar disc diffusion method along with qualitative analysis for presence/absence of phytochemicals. Antioxidant activity was measured by DPPH assay and the cytotoxicity was evaluated using MCF-7 (human breast adenocarcinoma) cancer cell. Presence of phytochemicals like alkaloids, flavonoids, reducing sugars and saponins were detected in the plant extract. The extract was found to show some level of antimicrobial activity against Staphylococcus aureus, Escherichia coli and Bacillus thuringiensis at 50, 100 and 200 mg/ml. The IC50 value of the extract was found to be 7.68 μg/ml. The extracts of Agave americana showed 50 % cell-death of MCF-7 in 12 h at 5 μg/ml. Although this study provided some scientific evidence for the medicinal value of Agave americana, further studies are still needed for the detailed evaluations of every molecule present in this plant along with screening in larger geographical area of Nepal
Hepatitis B and C Virus Infections among Blood Donors in Blood Transfusion Center, Pokhara, Nepal: Seroprevalence and its Associated Risk Factors
Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) infections lead to chronic diseases and are the most common causes of liver cirrhosis and cancer in developing countries like Nepal. The study is carried out to determine the seroprevalence of HBV and HCV by using a Rapid kit method and Elisa Method to find out its risk factors. The cross-sectional study was done among blood donating people from 16th August 2016 to 19th November 2016. Blood donors in Pokhara Valley were screened for anti-HCV antibodies, anti-HBV antibodies using third generation ELISA kits and automated ELISA Processor in serology laboratory at Central Blood Transfusion Service (CBTS) of Nepal Red Cross Society (NRCS) in Pokhara, Nepal. 1777 (87.2%) units were male blood donors and 260 (12.6%) units were female donors out of 2037 participants. Gender wise, the ratio between male and female was 1:0.1. HBV and HCV infection rate in blood donors were detected at 0.7% (15/2037) and 0.5% (8/2037) respectively. HBV infection rate in volunteer blood donor people was 0.7% (14/1881) which was higher than the replacement donors i.e. 0.6% (1/156). Similarly, in HCV infection rate in volunteer donor were 0.4% (8/1881). HBV infected people are detected higher than the HCV infected people among the blood donors. In addition, there was no significant relationship between positive results of HBV and HCV tests with the gender, age, tattoo, donor type
Potential of Ayurvedic Drugs in Differentiating Neuronal Stem Cells from Human Breast Milk: A Review
Recently, Stem cell therapy has revolutionized excellent clinical therapy with the potential of stem cells to differentiate into various cell types and it may help to replace different cell lines of an organism. Frequent, clinical trials are carried out to merge the new scientific stem cell information and techniques with traditional knowledge and plant extracts that may result in less toxic, affordable, and highly available natural alternative therapeutics. Ayurveda, the traditional Indian system of medicine has given great emphasis to the promotion of health. Ayurveda therapies are based on the restoration of body balance and nourishment of dhatus. Rasayana concept of Ayurveda explains tissue regeneration and cell renewal. Rasayana drugs and therapies provide research opportunities for the biology of regeneration. Specific Medhya Rasayana stimulates and nourishes respective Medha (Dhi, Driti, and Smriti) dhatus. Interpretation of this description offers clues for specific differentiation of neuronal stem cells from human breast milk (HBM) in the presence of some herbal extracts. The previous studies suggest that neuronal stem cells differentiate from Human Breast Milk (Human mesenchymal stem cell) more effectively with Madhya Rasayana drugs.
The present review highlights the potential of Ayurveda and its possible contributions in regenerative medicine
Development of Cost Optimized Horizontal Gel Electrophoresis Running Unit for Developing Countries (Nepal)
The use of expensive lab techniques has left many high schools and even university students unacquainted with the basic experimental procedures and protocols in developing country including Nepal. Horizontal gel electrophoresis is one of the expensive protocols, which every student in the laboratory may not get an equal chance to access individually. However, this technique, being indispensable and inevitable in molecular biology principles, is of abounding importance for students to be familiar with. Thus, realizing its importance, we present an extremely simple and inexpensive design of gel-electrophoresis unit, which emulates electrophoresis analysis with the use of nichrome and aluminum wires as a substitute for platinum wires, together with daily used plastic materials. Because of these factors, the approximate cost of unit design is significantly reduced to an amount of 10 USD. The efficiency of the substitute wires was confirmed and it resulted in satisfactory data characterized by good resolution of the DNA fragments. The inexpensive nature, good results and simplicity of the device make it an ideal unit for teaching and learning in developing countries