Journal of Nepal Biotechnology Association <p>The Journal of Nepal Biotechnology Association (J. Nepal Biotech. Assoc.) is an annual scientific publication of the Nepal Biotechnology Association. It is a double-blind peer-reviewed journal that publishes articles on different fields of Biotechnology.</p> en-US (Dr. Bijaya Pant) (Sioux Cumming) Thu, 14 Mar 2024 08:51:19 +0000 OJS 60 Exploring Holistic Wellness: Unveiling the Probiotic Wonders of Fermented Dairy – Meet Kefir and Its Health Benefits <p>This article explores the rich historical background and emerging health advantages of kefir, a fermented dairy product renowned for its abundant probiotics and positive impact on gut health and overall well-being. Additionally, the research delves into the microbial composition of kefir, its potential probiotic benefits, and the intricate interplay of microorganisms within the kefir grains. Examining specific <em>Lactobacillus</em> strains, the study assesses their health-promoting benefits, ranging from improved digestion and immune system support to potential contributions to mental health. By scrutinizing the probiotic microorganisms found in kefir, this paper aims to offer valuable insights to researchers, food technologists, and entrepreneurs interested in the nexus of nutrition, health, and sustainable food production.</p> Sushmita Soni, Sarbesh Das Dangol, Jarina Joshi Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 The Medicinal Fungus Cordyceps militaris: Academia to Industry in Thailand <p>The medicinal mushroom (<em>Cordyceps</em> species) is a highly valued source of useful natural products having diverse biological activities. <em>Cordyceps militaris</em> (commonly known as orange caterpillar fungus), a kind of herbal drug and food additive mushroom, were used as a tonic food in Asia from ancient times. Their products have been developed from cultivated fruit bodies and fermented mycelia of <em>C. militaris</em>. Various studies of culture techniques will be expected to reduce contamination. A traditional methodology (autoclave) to sterilize culture media was compared with a protocol that included hydrogen peroxide to sterilize culture media (without autoclaving and inoculating cultures without the laminar airflow cabinet) for growing <em>C. militaris</em> CMRU strain. The results showed that the two methods had no significant differences in cordycepin production and biomass production. Production of <em>C. militaris</em> products is now becoming a large industry in Thailand.</p> Tanya Tapingkae, Mongkol Yachai, Wanaporn Tapingkae Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Qualitative Analysis of Adulterant Mixed in Different Food Stuffs <p>This study focuses on identifying adulterants present in items such as food, fuels, chemicals and cosmetics, known for degrading their overall quality. The escalating concern over food adulteration prompted this research, emphasizing the detection of adulterants in daily consumables. The detrimental effects of food adulteration are profound, leading to health issues such as cancers (colon and peptic ulcer diseases), chronic liver diseases, electrolyte imbalance, kidney failure, heart diseases, blood disorders, and bone marrow abnormalities. The primary objective of this research is to ensure the quality of commonly consumed food items by detecting potential adulterants. Numerous rapid detection techniques have been developed to address this problem, including the implementation of quick and straightforward DART methods (Detect Adulterant Rapid Test). In this study, we applied various DART and DIY methods to test selected food items like milk, turmeric powder, and chilli powder. Each sample underwent testing with specific chemical reagents to determine the presence of adulterants. Post-tests, the samples were analyzed for observable changes, and conclusions were drawn regarding the presence or absence of adulterants in each tested item.</p> Prabhat Kharel, Anita Sapkota, Pallavee Regmi, Bimala Subba, Khaga Raj Sharma Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Ex-situ Conservation of Bulbophyllum leopardinum, A Threatened Medicinal Orchid of Nepal <p>A successful micropropagation method was developed via the in-vitro seed germination and seedling growth of the epiphytic and/or lithophytic orchid <em>Bulbophyllum leopardinum</em>, a species having horticultural and therapeutic significance. To enhance seed germination, several quantities and combinations of naphthalene acetic acid (NAA), 6-benzyl amino purine (BAP), indole acetic acid (IAA), gibberellic acid (GA<sub>3</sub>), and coconut water (CW) were added to 0.8% (w/v) agar-solidified MS medium. Half-strength MS medium has been experimented with alone and in combination with BAP, Kinetin (Kn), and GA<sub>3</sub> to promote shoot development. In-vitro-developed healthy shoots were chosen to establish roots in a half-strength MS (HMS) medium supplemented with various auxins. The best and earliest seed germination with the greenest protocorms (96.3±0.5% in 7 weeks) was achieved on HMS medium fortified with 15% CW (H15C). Further tests for the shoot as well as root development were continued with an H15C medium. H15C with 1 mg/l BAP and 1.5 mg/l Kinetin was most effective for early in vitro development and differentiation into seedlings with the many long shoots (9.3±0.1 shoots and 2.4±0.1 cm per culture) within 12 weeks of sub-culture. The most suitable rooting hormone was 1 mg/l NAA (4.2±0.26 roots per culture). This medium also produced the longest roots (1.9±0.09 cm per culture). By successfully developing a protocol for the mass propagation of <em>B. leopardinum</em>, this research has enhanced both the cultivation and the commercialization potential of this species.</p> Bir Bahadur Thapa, Krishna Chand, Laxmi Sen Thakuri, Manju Kanu Baniya, Bijaya Pant Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Induction, Proliferation and Differentiation of Callus in Paris polyphylla Sm. through Leaf Culture <p><em>Paris polyphylla</em> Sm. is a vulnerable medicinal plant employed in the treatment of various ailments. This study seeks to establish a protocol for callus induction, proliferation, and differentiation of <em>P. polyphylla</em>. Immature leaf explants were cultured on MS medium with varying concentrations of plant growth regulators (PGRs), including 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (Kn), 6-benzylaminopurine (BAP), Thidiazuron (TDZ), α-Naphthalene acetic acid (NAA), and Gibberellic acid (GA<sub>3</sub>), along with 10% coconut water. After 12 weeks of primary culture, the optimal callus induction was observed in MS medium supplemented with 0.25 mg/l 2,4-D + 0.5 mg/l Kn. In the secondary culture at 8 weeks, the best callus proliferation, as determined by callus weight or growth index, occurred in MS medium supplemented with 2.0 mg/l BAP alone, 2.0 mg/l Kn alone, 1.0 mg/l TDZ alone, combinations of 2.0 mg/l Kn + 1.0 mg/l BAP + 2.0 mg/l GA<sub>3</sub>, and combinations of 0.5 mg/l NAA + 2.0 mg/l BAP + 2.0 mg/l GA<sub>3</sub>, as well as 10% coconut water. Furthermore, callus differentiation into mini rhizomes with root primordia was successfully achieved in MS media containing 2.5 mg/l Kn and 10% coconut water. This study reports, for the first time, the formation and differentiation of callus from leaf explants in <em>P. polyphylla</em>. Large-scale callus generation from leaf explants has the potential to enhance the production of bioactive secondary metabolites for therapeutic purposes and facilitate the development of plantlets through organogenesis.</p> Chandra Bahadur Thapa, Krishna Kumar Pant, Hari Datta Bhattarai, Mahendra Thapa, Bijaya Pant Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Chemical and Morphological Characterization of Crinis Carbonisatus <p>Crinis Carbonisatus, prepared by pyrolysis of human hair, is known as a traditional Chinese medicine used for increasing blood clotting and wound healing. Its uses have been explored in literature but no detailed structural study is yet reported. This work is aimed at studying the chemical and morphological variation of Crinis Carbonisatus under given heating conditions. Crinis Carbonisatus was obtained after pyrolyzed of human hair at 300 °C in a sealed ceramic pot. The obtained samples were characterized in terms of their physicochemical properties by scanning electron microscopy (SEM), Fourier Transform Infrared (FTIR) spectroscopy and X-ray Diffraction (XRD). Distinct morphology with nanoparticulate structure was observed on the SEM micrograph. FTIR spectroscopy of the samples revealed the presence of functional groups like –OH, -COO<sup>-­</sup>, and -NH as well as methyl (-CH<sub>3</sub>) and methylene (-CH<sub>2­</sub>-) groups. The nanoparticulate graphitic form was confirmed by XRD. It has been found that with the increase in pyrolysis time; the amorphous nature of the Crinis Carbonisatus materials increases while their particle size decreases.</p> Tika Ram Bhandari, Bidit Lamsal, Prasamsha Panta, Nilam Shrestha Pradhan, Marco Liebscher, Tika Bahadur Katuwal, Rameshwar Adhikari Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Pharmacological Activities of Six Species of Hedychium J. Koenig from Nepal <p>Species of <em>Hedychium </em>Koenig are perennial herbs. Some of the species like <em>H</em>. <em>ellipticum</em>, <em>H. spicatum</em>, <em>H</em>. <em>coronarium</em> are traditionally used as medicinal plants. In the present study, methanolic rhizome extracts of six different species of <em>Hedychium</em> namely <em>H</em>.<em> spicatum</em>, <em>H</em>. <em>ellipticum</em>, <em>H</em>.<em> thyrsiforme</em>, <em>H</em>.<em> coccineum</em>, <em>H</em>.<em> gardnerianum</em> and <em>H</em>. <em>coronarium</em> were analysed for their antioxidant, antidiabetic and antibacterial activities. The antioxidant activity analysed by DPPH assay showed the highest potential (lowest IC<sub>50 </sub>value) in <em>H</em>.<em> coccineum </em>(148.82±2.83 µg/ml) and lowest potential (maximum IC<sub>50 </sub>value) in <em>H</em>. <em>thyrsiforme </em>(996.55±9.42 µg/ml). The rhizome extracts of different species showed moderate α-amylase inhibition activity <em>in vitro</em>. The highest α-amylase inhibition (79.67%) was observed for <em>H. coronarium </em>while the lowest inhibition (64.0%) was observed in<em> H. thyrsiforme.</em>&nbsp; However, these values were found lower than the value (92.37%) obtained for positive control, i.e., Acarbose. The antibacterial activity was determined against two Gram-positive (<em>Bacillus subtilis</em> and <em>Staphylococcus aureus</em>) and two Gram-negative (<em>Klebsiella pneumoniae</em> and <em>Pseudomonas aeruginosa</em>) bacterial strains by agar well diffusion method. Except for <em>H. ellipticum</em> the extracts of all other species showed antibacterial activity against all the bacterial strains tested. The extracts of <em>H. ellipticum </em>showed antibacterial activity only against <em>B</em>.<em> subtilis</em> and <em>K</em>.<em> pneumoniae</em>. The extract of <em>H. coronarium</em> showed the highest zone of inhibition (16.67±1.15 mm) against <em>B</em>.<em> subtilis</em>. However, the antibacterial activity was weak compared to standard antibiotics for all the extracts and at all concentration tested. These results show that rhizomes of other species can also be used in the same manner as that of <em>H</em>.<em> coronarium</em> and <em>H</em>.<em> spicatum</em>, two species most used in various ethnomedicinal applications.</p> Dama Pun, Giri Prasad Joshi, Deepak Raj Pant Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 In vitro Callus Regeneration and Chlorophyll Content Estimation in Glycine max (L.) variety from Uttarakhand, India <p>Soybean (<em>Glycine max) </em>is considered one of the most substantial produced crops across the globe because of its high nutritional value. It is a great alternative for lactose-intolerant patients and vegans to fulfil their daily protein requirements. Soybean has various varieties depending upon the colour of their seed coat. In the past few years, the consumption of Soybean has increased which demands higher production, better yield, and better seed quality. Conventional propagation methods fail to fulfil such demands. The alternate method of plant tissue culture ensures rapid mass propagation, better yield, and quality of the plants. However, the technique is often beset with challenges of low-field performance of tissue culture-raised plants due to defective chloroplast machinery. The present study investigates the effect of various plant growth regulators (PGRs) on <em>in vitro</em> propagation of soybean cultivars from different regions of Uttarakhand, India, and their effect on chlorophyll content in the regenerated tissues.</p> Aarushi Gautam, Priya Saini, Ayushi Negi, Astha Saini, Manu Pant Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Association between Vitamin D Receptor Fokl and Bsml Gene Polymorphism and Diabetes Mellitus in Nepalese Population <p>Vitamin D being involved in the secretion of insulin is a known fact. Moreover, studies have shown that steroids might be a factor in influencing insulin sensitivity. Vitamin D receptor (VDR), a factor required for genetic regulation involving vitamin D, thus can be regarded as a good candidate for Diabetes Mellitus (DM). Several studies have been conducted on the association between VDR polymorphism and the risk of DM but did not provide clear-cut answers. This study was conducted to search for the involvement of FokI and BsmI polymorphisms of the VDR gene with DM in a Nepalese population. A total of 200 blood samples were collected; 100 from clinically diagnosed DM patients and 100 from healthy controls. DNA was extracted from blood by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, where FokI and BsmI primers as well as restriction enzymes were used. After restriction digestion, SNPs of FokI (T/C) [rs2228570] and BsmI (A/G) [rs1544410] were assayed using agarose gel electrophoresis. As patients and controls were likened for genotype distribution and allelic frequencies, it was found that the frequency of VDR gene BsmI&nbsp;rs1544410 differed significantly (p &lt; 0.05, each) between cases and control whereas A allele was dominant (91%) in healthy controls with Odd ratio (OR) of 0.55, unlike VDR Fokl were not significantly associated between subjects and control. The data obtained from this research suggests that the VDR gene (especially BsmI) is associated with the risk of DM.</p> Anil Kumar Sah, Melina Dahal, Sandip Baniya, Santoshi Pyakurel, Sonu Rai, Lasta Maharjan, Susmita Bista, Keshab Raj Budha, Biraj Lohani, Prakash Poudel, Pragati Poudyel, Bajrangi Rauniyar Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Oral Microbial Diversity Among Nepalese Individuals Across Various Geographical Regions <p>A comprehensive study encompassed the collection of 153 samples derived from oral patients across 17 diverse locations throughout Nepal. The assortment of samples included extracted teeth, dental plaque, and dental calculus, procured from dental clinics, dental hospitals, and dental camps. Employing six distinct culture media, namely nutrient agar (NA), Muller Hilton agar (MHA), mannitol salt agar (MSA), blood agar (BA), brain heart infusion agar (BHA), and potato dextrose agar (PDA) for potential fungal strains, plates were meticulously incubated at 37°C for 5-7 days. The ensuing bacterial colonies were judiciously isolated, and their morphological and biochemical traits were scrutinized. The microscopic structures of the bacterial cells were examined, considering shape, size, colour, opacity, and texture. Gram-staining was employed, and each colony's biochemical attributes were assessed for protease, pectinase, cellulase, and lipase enzymes. From the 1200 colonies isolated from dental samples, 300 diverse colonies, distinguished by morphological and biochemical characteristics, were chosen for further taxonomic identification. Subsequent sequencing revealed the identification of 60 distinct species within 21 genera of bacterial isolates, including <em>Achromobacter</em>, <em>Bacillus</em>, <em>Chryseobacterium</em>, <em>Citrobacter</em>, <em>Curtobacterium</em>, <em>Enterobacter</em>, <em>Enterococcus</em>, <em>Escherichia</em>, <em>Flavobacterium</em>, <em>Klebsiella</em>, <em>Kocuria</em>, <em>Lyinibacillus</em>, <em>Novosphingobium</em>, <em>Ochrobactrum</em>, <em>Proteus</em>, <em>Pseudomonas</em>, <em>Sporosarcina</em>, <em>Staphylococcus</em>, <em>Stenotrophomonas</em>, <em>Serratia</em> and <em>Streptococcus</em>. The research underscored the presence of various pathogenic bacterial species in oral samples.</p> Nirmal Panthi, Mukesh Thapa, Rajani Malla, Arun Nagarkoti, Hari Datta Bhattarai, Babita Paudel Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000 Impact of Invasive Ageratina adenophora on Soil Fungi in Native Plant-Grown Soils in Nepal <p>Invasion of alien plants impacts both the above- and below-ground communities. The specific response of soil microbes to alien plants may depend upon the presence of native inhabitants and the origin of the byproducts released from different parts of invasive plants. This study evaluated the response of common soil fungi against <em>Ageratina adenophora </em>byproducts (extracts) in the presence of two native shrubs, <em>Elsholtzia blanda</em> and <em>Osbeckia stellata</em>, in Nepal. A total of eight fungal species were isolated from soils where these two native species were grown separately, and the occurrence of the fungi was evaluated. The occurrence and frequency of fungus species varied with extracts of <em>A. adenophora </em>leaves, litter and roots as well as with the presence of specific native plants. Particularly, <em>A. adenophora </em>further inhibits the fungi that are naturally less frequent in soil, like <em>Hormodendrum </em>sp. and fresh leaves and litter of <em>A. adenophora </em>were responsible for inhibiting antagonistic fungi like <em>Trichoderma harzianum.</em> Influence in below-ground fungal communities by <em>A. adenophora</em> is one of the reasons for poor growth and development of native seedlings and the mechanism could be a strategy of invasion of <em>A. adenohora </em>in novel areas.</p> Tej Bahadur Darji, Seeta Pathak, Reetu Deuba, Khageshwari Saud, Gunanand Pant, Lal Bahadur Thapa Copyright (c) 2024 Nepal Biotechnology Association Thu, 14 Mar 2024 00:00:00 +0000