International Journal of Applied Sciences and Biotechnology

367-372

Authors: Bishnu Prasad Neupane, Jiban Shrestha

This review paper highlights scenario of entomological research in grain legumes in Nepal into headings of the monitoring, survey and surveillance, insect pest management, existing problem and future strategies. A survey study on yield loss assessment of chickpea due to Helicoverpa pod borer under field condition has been recorded up to 75% in the year 1996-1998 at Banke and Bardiya district. While monitored Helicoverpa armigera through pheromone trap, during the 2nd week of March, peak catches of 91 male moths were recorded at Rampur, while it was the maximum (42 male moths) during the 3rd week of March. Synthetic pyrethroids, deltamethrin, fenvalerate and cypermethrin used at 0.01% concentration (a.i.), were effective in controlling the chickpea pod borer, Helicoverpa armigera Hubner. Genotypes NCH-18, NCH -31, NCH-138 and ICC 3075 WR were found resistant. Ripcord 10%@0.05% was found to be superior and effective chemical insecticides to control stem fly and soybean hairy caterpillar. The late sowing date December, 10 showed significantly higher percentage of chickpea pod borer damage with lower grain yield. Inter crops combinations of Chickpea +wheat (2:1) ratio was found effective against Helicoverpa armigera damage (1.2%) followed by chickpea+barley (2.36%) and Chickpea+linseed (2:1) ratio (3.7%). Bakaino (Melia azederach L.) was identified as the best indigenous pesticides to check the normal growth of hairy caterpillar (Spilarctia casigneta kollar) under laboratory condition. Metarhizium anisopliae was found effective for the management of Helicoverpa armigera that caused maximum mortality (94.67%), infection (45.33%) within 7.49 days (LT50). Similarly, Beauveria bassiana caused maximum mortality (98.67%), infection (80.00%) with the least LT50 value (5 days). Crop sprayed with HaNPV had the lowest pod damage (0.3%) and the highest in Racer (2.2%). This review work provided a lot of information to conduct entomological research activity conducted in grain legumes and opens the door for future research and strategy.

381-386

Authors: Jignasha G Patel, J.I. Nirmal Kumar, Shamiyan R Khan

The study was aimed to determine the chronic toxicity of Polynuclear aromatic hydrocarbon – Anthracene in response to pigments and metabolic study on three different cyanobacterial species such as Synechocystis sp., Anabaena fertilissima, and Nostoc muscorum. Test organisms were treated at different doses and encountered LC50/Mean Lethal Concentration (at which 50% lethality/ growth reduction occur) separately at 7.0 ppm for Synechocystis sp, 5.0 ppm for Anabaena fertilissima and 1.5 ppm for Nostoc muscorum. The influence of anthracene on pigments, metabolites and enzymes was carried out. The test doses caused concentration dependent and decreased pigments like carotenoids and phycobilliproteins. Depletion of carbohydrate by 65 to 80% and proteins by 58 to 78% was encountered with rise in Anthracene concentrations after 16th day exposure in case of Synechocystis sp however, phenols were found to raise by 26 to 37% with increased anthracene concentrations. Similar trend also observed in other two tested blue green algae. Thus the Synechocystis sp.is more tolerant to anthracene treatments as compare to Anabaena fertillissima but Nostoc muscorum showed highest sensitivity to anthracene.

392-397

Authors: Hriush Adhikari, Sangam Ghimire, Binod Khatri, Yuvraj K.C.

Background and Aim: In Nepal not much of study of Thermophilic area and Thermophiles have been done. Thermophilic bacteria are less studied but are important group of microorganisms due to their ability to produce industrially important enzymes. Methods: In this study, thermophilic bacteria were isolated from hot spring of Bhurung, Nepal. Wide range of bacteria that could grow at high temperatures and tolerate extreme temperature were characterized by morphology, biochemistry and sequencing of its 16S rRNA gene sequence. The isolates were screened for production of extracellular enzymes like protease, amylase, lipase, cellulase, caseinase, pectinase and xylanase activity. Phylogenetic tree construction and G+C content evaluation of the isolate was also studied. Results: 15 isolates with ability to tolerate high temperatures were identified as Bacillus sp. by morphology, biochemistry and sequencing of its 16S rRNA gene sequence. BLAST search analysis of the sequence was performed and result showed maximum identity (99% similarity) with Bacillus licheniformis, Bacillus subtilis and Bacillus pumilus. Isolated strains exhibited considerable amount of extracellular exozymes activity. Phylogenetic analysis of the isolates revealed the relatedness among the species. The G+C content of each species was also evaluated and was found to be in range of 54.87 to 55.54%. Conclusion: The study of isolates confirmed that the isolated Bacillus sp. to be a true thermophile and could be a source of various thermostable exozymes which can be exploited for pharmaceutical and industrials applications. Much detailed study of the isolates can

402-407

Authors: Dipak Kathayat, Swoyam P. Shrestha, Santosh Dhakal, Laxman Ghimire, Mahesh K.C.

A cross-sectional study was conducted on four major pig raising districts of eastern and mid-western region of Nepal from February to May 2014 to find out the seroprevalence of Trichinella spp. A total of 184 serum samples of pigs were collected and antibodies against trichinella were detected using ID screen trichinella indirect multi- species ELISA kit. The study revealed prevalence rate of 3.8% and difference in prevalence rate according to age, sex, breed, rearing system, ecozone, region & district were statistically insignificant (P>0.05) as analysed by Chi-square test using PHStat version 2.5 and Fisher’s exact test. This study confirms that antibodies of Trichinella spp. are circulating in pigs of Nepal. Further, the knowledge, attitude and practices survey of meat borne helminthic zoonoses was conducted among 50 pig raisers and pork consumers by face to face interview using a semi-structure questionnaire. This survey concludes that although there were significant portion of the respondents aware of meat borne helminthic zoonoses but there were still a noticeable proportion of respondents who didn’t have a proper knowledge that upsurge public health risks. Moreover, present-day situation of their pig raising practices & pork consumption system possess them to a menace of public health zoonoses. be done for enhancing the potential of isolates.

417-422

Authors: Hari Kumar Prasai, Jiban Shrestha

Coordinated Varietal Trial (CVT) and Advanced Varietal Trial (AVT) of wheat were conducted at Regional Agricultural Research Station, Doti during the year 2012 and 2013. Microplot Yield Trial (MPYT) were conducted during the year 2013. Total 20 genotypes were included in CVT experiment of both years. Although the difference in grain yield due to genotypes was not found significant during the year 2012, NL 1144 recorded the highest grain yield (4309 kg/ha) followed by NL 1140 (4295 kg/ha) and NL 1147 (4165 kg/ha) respectively. But in the year 2013, NL 1097 produced the highest grain yield (4641 kg/ha) followed by NL 1135 (4383 kg/ha) and NL 1164 (4283 kg/ha) respectively. Statistically, the difference in grain yield due to genotypes was not found significant in the year 2013. Combined analysis over years was also carried out. Out of 20, only 10 genotypes were included in the CVT experiment, which were found similar in both years. Genotypes NL 1097 (4079 kg/ha), NL 1140 (3814 kg/ha) and NL 1093 (3773 kg/ha) were found high yielding genotypes for river basin agro-environment of far western hills. Statistically, effect of year in tested characters was found significant whereas treatment effect was observed non-significant. Similarly, 20 genotypes of wheat were included in AVT of wheat during the year 2012 and 2013. Out of the genotypes included in AVT during the year 2012, KISKADEE No.1recorded the highest grain yield (3824 kg/ha) followed by CHEWINK No. 1 (3643 kg/ha) and WK 2120 (3583 kg/ha). Statistically all the tested characters except grain yield were found significantly different due to genotypes. But in the same experiment of the year 2013, WK 2412 genotype recorded the highest grain yield (4407 kg/ha) followed by WK 2411 (4329 kg/ha) and Munal-1 (4054 kg/ha). Statistically the difference in grain yield and other tested characters were found significantly different. Due to dissimilarity in the tested genotypes we could not carry-out the combined analysis over years. Total 30 genotypes were included in the MPYT experiment of the year 2013. Genotype WK 2272 recorded the highest grain yield (6080 kg/ha) followed by the genotypes WK 2274 (5152 kg/ha) and WK 2278 (4480 kg/ha) respectively. Statistically, the difference in grain yield and other tested characters were found significantly different due

427-430

Authors: Moti Lal, Shiv Kumar Verma, Vipin Mishra, Mira Debnath

An amylolytic strain was collected from rotten potato and its activity evaluated. The isolated strain was cultivated for amylase production in shake flasks at 35±2oC and the fermentation pattern was studied. Optimum temperature for maximum enzyme synthesis was observed at 35°C, when initial pH of fermentation medium was adjusted to 5.5. Maximum extracellular amylase activity of 7.9 U/mL and the maximum intracellular activity of 320 U/mL was recorded. Although maximum biomass was present at 12.6 g/L but highest growth rate was observed between 08 to 40h with maximum at 36h. The extracellular amylase present in the broth was partially purified with an overall yield of 44% through purification procedure of ammonium sulphate precipitation. After completed extraction and partial purification and stabilization, the stability of enzyme was observed in a range of temperature and pH between 60°C-90°C and 2-8 pH respectively. Maximum enzyme activity was demonstrated at 90°C, and pH of 5.5 and 6.5. The thermo stability of the amylases of this Bacillus species was comparable to that of amylases from other microbial sources.

434-438

Authors: Mrinalini Damle, Nilam Dalavi

Ellagic and Gallic acid are main phytoconstituents of S.cumini seeds. These are the phenolic compounds. An approach for the stress degradation was successfully applied for the development of stability indicating HPLC method for the determination of Ellagic and Gallic acid. Sample was resolved on a Hypersil C18 (250*4.6 mm particle size 5?) column. The mobile phase consisted of 1% OPA and ACN and in the ratio of 70:30 v/v which was sonicated to degas and delivered at a flow rate of 1ml/min at ambient temperature. The retention time of Ellagic acid and Gallic acid was 3.1±0.05 & 4.1±0.05 minutes. Studies were performed using an HPLC system equipped with a UV detector; the response was monitored at 271nm. The method is specific to Ellagic and Gallic acid; it is able to resolve the peak from ethanolic extract of s.cumini seeds and formulation. The calibration curve was linear over the concentration range of 8-24 ?g/ml (r2=0.997, 0.998 resp). The limit of detection for Ellagic acid and Gallic acid was found to be 0.25?g/ml, 0.15?g/ml resp. and the quantification limit was about 0.75?g/m, 0.49?g/ml. The accuracy of the method was established based on the recovery studies. The markers were subjected to acid, base, neutral hydrolysis, oxidation, thermal degradation and photolysis. The method was successfully validated according to ICH guidelines Q2 (R1).

443-445

Authors: M. Poudel, H.K. Paudel, B.P. Yadav

The research was conducted under RCBD with three replication to study the inter relationships among traits affecting yield and its affecting character at National Maize Research Programme (NMRP), Rampur, Nepal in 2013. The eleven inbreeds line of maize variety at winter season in 2013 were evaluated for estimation yield and its affecting character. For efficient selection of grain yield, it is necessary to know relation of yield of maize and its morphological traits which are influencing on the grain yield. One of the objectives of this paper was to determine interrelationship between traits and the yields. Correlation coefficients among traits showed that yield was positively and highly significantly related with number of kernel rows per ear (0.788), number of kernels per row(0.571), ear girth(0.516), plant height(0.498), hundreds kernels weights(0.444) and significantly related with other character. .Number of Kernels rows per ear, number of kernels per row, plant height could be the important selection criteria in improving open pollinated maize varieties for high grain yield.

452-458

Authors: Attiya Mohamedin, Magdy Michel, Marwa Tolba

Ninety ?Koshari? meals samples were collected from some restaurants and street vendors located in six public quarters in central Cairo, to determine the microbiological quality of them directly after cooking and preparing and after 4 & 8 hrs of storage at room temperature. These samples examined for aerobic bacteria, Esherichia coli and coagulase positive Staphylococci to conduct a preliminary microbial risk assessment for them in” koshari “meals. According to the CDPH (2009) only 53 samples (59%) were of satisfactory microbiological quality for Aerobic plate count (APC) and 81 samples (90%) were positive for E. coli cells and 58% of them (47 samples) are acceptable quality. About coagulase positive S. aureus, 28 samples (31.1 %) were positive and only 60.7% of them (17 samples) of satisfactory microbiological quality.Moreover, the percentage of unacceptable microbiological quality samples tested (potentially hazardous) reached to 36 samples (40%), 29 samples (32.2%), and 17 samples (18.9%) for APC, E. coli and S. aureus respectively, after 8 hrs of storage at room temperature. This study reveals that “Koshari” meals sold on the public areas are unwholesome and could be a potential source of food-borne bacteria pathogens if not properly handled. Option might be to suggest that the product should be consumed within short time of purchase in these places. Improvements in processing and handling are required and the need of food-borne bacteria disease surveillance indicated. In addition, it was evident that the Egyptian Food Code needed new legal revisions.

464-473

Authors: Suresh Kumar Wagle, Neeta Pradhan, Madhav Kumar Shrestha

Asala or snow trout (Schizothorax richardsonii, Cyprinidae), one of highly valued freshwater fish of Transhimalayan regions, is distributed in upper reaches of all major river systems of Nepal. Morphometric diversification between six river populations of S. richardsonii was examined to identify intraspecific unit for enabling better management of the resources. Significant differences were observed in 17 measured morphometric characters of 207 specimens among the six river populations. Multivariate analysis of variance (Wilks' test) indicated a significant difference for mean vectors of mophometric measurements (? =0.012, F85, 731 = 19.999, P<0.0001) among populations. Principal component and discriminant functions (DFs) analysis of morphometric measurements revealed high seperation of the stocks. The analysis showed that most of the shape and size variation among these populations occurs in the head region, body depth and fin length. Apparent morphometric divergence among S. richardsonii samples showed the existence of three differentiated groups viz., the Indrawati and Khudi populations, the Melamchi and Phalaku Rivers, and the Sabha and Tadi River populations of Nepal. The results of this study may be useful in fisheries management and potential exploitation of this species in coldwater aquaculture.

479-482

Authors: Aruna C. Shanmugavadivu, T. Deepika, R. Hemamalini, G. Madhusudhanan, K. Indhu, N.C. Ramesh Babu

The p53 expression on Gliomas are used for subtyping of tumours. The samples were extracted from the paraffin embedded tissue blocks. The Grade IV and Grade II were tested by IHC (ImmunoHistoChemistry) staining procedures. The primary antibody used was p53. It was found that there is a high expression of p53 in Grade IV (82%) was Score 3 and low expression p53 in Grade II (18%) in which the score was 1. This is in contrast to previous studies of subtyping of Gliomas where, classical types have low expression of p53 and proneural type have high expression of p53. This subtyping was based on score 0 - 3. Score 0 has no positive nuclei, Score 1 has 10% positive nuclei, Score 2 has 10 -30% positive nuclei while score 3 has >30% of positive nucleus.

490-503

Authors: Ravi Gedela, Naga Sai Babu Makke, Dinesh Karra

We have studied insilico on evolutionary uniqueness of phytoene synthase, which is one of the regulatory enzymes of ?-carotene synthesis in Neurospora crassa. This study reveals multiple sequence alignments showed high sequences with similarity within a species of bacteria, fungi and higher plants. This results designate interestingly between species of bacteria-fungi, fungi-plant, and among the species of bacteria-fungi-plant, showed tremendously less sequence with similarity, except bacteria-plant (high sequence with similarity) respectively. In Phylogenetics tree analysis showed within species of bacteria, fungi and plant 91%, 92% and 99% homology. Whereas in between species of bacteria-fungi, bacteria-plant, fungi-plant, and among the species bacteria-fungi-plant showed 99%, 96%, 100%, and 91%-99% homology respectively. N. crassa phytoene synthase enzyme encode (Isoprenoid Biosynthesis enzymes, Class 1) protein size 610aa, Cyanobacteria phytoene encode (Isoprenoid Biosynthesis enzymes, Class 1) protein size 310aa, and Oryza sativa Indica phytoene synthase 1 (chloroplast), (Isoprenoid Biosynthesis enzymes, Class 1) encode protein size 421aa (e- value 0.0, 0.0 and 0.0; identity 100%, 100% and 100%; Max.score:1238, 644 and 870) respectively. We studied insilico on basis of an evolutionary Endosymbiotic theory; a bacterium is the ancestors to eukaryotes.

513-519

Authors: Binod Prasad Luitel, Bhim Bahadur Khatri, Duryodhan Choudhary, Bishnu Prasad Paudel, Sung Jung-Sook, On- Sook Hur, Hyung Jin Baek, Ko Ho Cheol, Ryu Kyoung Yul

This experiment was conducted to assess the genotypic variation for the growth, yield and yield components of potato grown under drought and irrigated conditions at Hattiban Research Farm, Khumaltar during the summer cropping season (Feb.-May) of the two consecutive years (2013 and 2014). The results revealed that canopy cover, stem height and number of leavers were more sensitive to drought and the effect was more pronounced in early cultivar Desiree. Stem height and leaf number of plant were more (36% and 45%, respectively) reduced in Desiree than other genotypes. Mean tuber number decreased by 55% under drought condition. Drought stress reduced marketable tuber number by 79% as compared to irrigated treatment. Drought stress reduced the marketable tuber yield from 70% to 87%. The clones CIP 392242.25 and LBr-40 had relatively lower yield loss, and less drought susceptibility index under drought indicating their tolerance to drought in field condition. Further experiment is recommended to study the physiological parameters of these genotypes under different water and soil conditions.

528-536

Authors: Yehia A. Osman, Waled M. El-Senousy, Adel A. El-Morsi, Mohammed K. Rashed

The fecal bacteria have been taken as the gold standard for water industry. However, the spread of viral gastroenteritis due to drinking water have given a momentum to a recent push by microbiologists to consider viruses as important pollution indicator as fecal bacteria. Therefore, we designed a study to evaluate the efficiency of two types of water purification systems: the traditional water treatment plant and two types compact units. Both systems produced drinking waters free of bacteria, chemical contaminants and mostly viruses free. However, recent advances in molecular biology techniques, such as RT-PCR have detected Rotaviruses in chlorinated drinking waters resulted from all systems. The frequency of Rotaviruses since October 2010 till September 2012 in Shark El-Mansoura WTP in drinking water samples was 12.5% similar to raw water. While the compact unit at Depo Awam (American design) the frequency of Rotavirus was 16.6% in both raw and drinking water samples. On the other hand the virus frequency in the raw and drinking water sample in El-Danabik unit (Egyptian design) were 12.5% and 4.16% respectively. Signifying failure of the chlorination process in removing viruses completely. However, detection of Rotavirus genome in the drinking water samples does not means the presence of its infectivity. The infectious ability of the rotaviruses was confirmed by CC-RT-PCR in all positive samples, where viral RNA was not detected in the collected drinking water samples. In conclusion RT-PCR and CC-RT-PCR techniques high lightened the need to include viruses as mandatory pollution indicator in water treatment plants.

541-544

Authors: A. Khanal

Plant nutrient is one of the limiting factors affecting crop production. Nitrogen and boron are major nutrients in case of broccoli. So, an experiment was carried out to evaluate the effect of nitrogen and boron in seed yield and yield attributing characters of broccoli in Rampur, Chitwan during winter season. The experiment was laid out in factorial RCBD design with four levels of nitrogen and two levels of boron. Each plot consists of 25 plants which were separated by 60 * 60 cm spacing. There are altogether eight treatments replicates thrice. Local variety Calabrese was used. Significant effect of different dose of nitrogen and boron on yield attributing characters was found. Also interactive effect of nitrogen and boron in number of pods, pod length, seed yield and number of seeds per pod was found significantly different.

552-560

Authors: Mohamed A.M. El-Awady, Mohamed M. Hassan, Yassin M. Al-Sodany

This study was designed to isolate and characterize endophytic and rhizospheric bacteria associated with the halophyte plant Sesuvium verrucosum, grown under extreme salinity soil in Jeddah, Saudi Arabia. The plant growth promotion activities of isolated bacterial were evaluated in vitro. A total of 19 salt tolerant endophytic and rhizospheric bacterial isolates were obtained and grouped into six according to genetic similarity based on RAPD data. These six isolates were identified by amplification and partial sequences of 16S rDNA as Enterobacter cancerogenus,Vibrio cholerae, Bacillus subtilis, Escherichia coli and two Enterobacter sp. Isolates were then grown until exponential growth phase to evaluate the atmospheric nitrogen fixation, phosphate solubilization, and production of phytohormones such as indole-3-acetic acid, as well as 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. While, All of the six strains were negative for ACC deaminaseactivity, two isolates showed Nitrogen fixation activity, three isolates produce the plant hormone (Indole acetic acid) and two isolates have the activity of solubiliztion of organic phosphate. Among the six isolates, the isolate (R3) from the soil around the roots is able to perform the three previous growth promoting possibilities together and it is ideal for use in promoting the growth of plants under the high salinity conditions. This isolate is candidate to prepare a friendly biofertelizer that can be used for the improvement of the crops performance under salinity conditions.