International Journal of Applied Sciences and Biotechnology

3-14

Authors: Shivani Sachdev, Sanjay Kumar Ojha, Snehasish Mishra

Amylase is one of the leading enzymes used in industry from decades. The preliminary function of this enzyme is the hydrolysis of the starch molecule into glucose units and oligosaccharides. Amylases have spectacular application in broad spectrum of industries such as food, detergent, pharmaceutical and fermentation industries. Among different type of amylases α- amylase is in utmost demand because of its striking features. This particular enzyme is a good substitute over the chemicals catalyst used in industries. α- amylases can be acquired from different sources such as microorganism, animals and plants. Microorganisms are the major source of production of amylase because of the ease of availability, manipulation and operation. The starch converting enzymes is basically generated using submerged fermentation. Some of the prominent characteristics of amylase are its mode of action, substrate specificity and operating condition (temperature and pH). Amylases from different bacterial sources contribute differently to the particular trait of the enzyme. Bacillus amylases have been studied and applied so far because of their robustness in nature and easy accessible pure form of it. Thus this makes it more specific and fit for distinct application in the industry. The purpose of this manuscript was the comparative analysis of the physical and chemical features of α amylases from Bacillus species. It also focuses on the unique characteristics of this enzyme and their industrial applications.

21-26

Authors: A.I. Inegbenebor, A.O. Inegbenebor, R.C. Mordi, N. Kalada, A. Falomo, P. Sanyaolu

Clay is one of the major raw materials in ceramics, paint and paper industries. In this paper, a report is given on the chemical composition of clay deposits in Ilaro and Ikorodu, South West Nigeria and their suitability as raw materials in the above mentioned industries is suggested by comparison of their composition with industry standards. The clay samples were activated with H2SO4 and their moisture content was determined. The mineral content of the clays was determined using atomic absorption spectroscopy, which showed significant presence of Si, Al, Fe, and Ti. The morphology of the clays was studied using scanning electron microscope (SEM/EDX) in combination with X-ray analysis. The investigation revealed the agglomeration of the particles of quartz and rutile on the surface structure of the activated clays. The results obtained from their observed properties and chemical compositions indicate that these clays are suitable for use in the ceramics, paint and paper industries. This proposition is supported by reduction in moisture contents of the activated clay samples from both Ilaro and Ikorodu.

32-38

Authors: Gowri Rajkumar, Jagathpriya Weerasena, Rangika Silva, Kumudu Fernando

Genetic relationships among 28 new improved rice varieties were established using Amplified Fragment Length Polymorphism (AFLP) markers. Cultivars were analyzed with 10 EcoR1 and MseI primer combinations. A total of 517 fluorescent AFLP markers were generated and analyzed. Of these 480 fragments were polymorphic (92.84%) and 37 (7.16%) fragments were monomorphic. The Jaccard’s similarity indices (J) based on the AFLP profiles of the 28 varieties were computed and Unweighted Pair Group Method with Arithmetic mean (UPGMA) based dendrogram was constructed. The dendrogram separated varieties into three major clusters. Outliers used in the study were uniquely separated from the rest confirming the reliability of data and analysis. The Cophenetic correlation with 0.862 strongly supported the clustering pattern of UPGMA dendrogram. Principal Coordinate analysis and the unrooted tree also confirmed the clustering pattern of the UPGMA dendrogram. Rice varieties in the same cluster showed similar characteristic features (Eg. Grain colour, life span etc). Therefore this genetic diversity data at molecular level will provide detailed estimates of the genetic variation among Sri Lankan new improved rice varieties and also useful in ex situ and in situ genetic conservation, utilization and exchange of genetic material.

44-52

Authors: K Rajendiran, L Gowsalya, J Sudaroli Sudha

Even though seeds form the natural progenies of all sexually reproducing plants, change in climate makes it mandatory to device in vitro methods for selecting germplasm for conservation and regeneration. An experiment was conducted to induct callus in stem explants (nodal region from third node from top of canopy) harvested on 30 days after seed germination from in situ control and supplementary ultraviolet-B irradiated (UV-B = 2 hours daily @ 12.2 kJ m-2 d-1; ambient = 10 kJ m-2 d-1) three varieties of cowpea (Vigna unguiculata (L.) Walp.) viz. Gowmathi, Fola and NS-634 to assess their response to in vitro culture. Axillary bud initiation occurred only from GOWMATHI control stem explants, while callus induction occurred in GOWMATHI and FOLA control stem explants. Stem explants harvested from all the three UV-B stressed cowpea varieties did not proliferate callus. The nodal stem explants of the three varieties of cowpea are not the suitable materials for germplasm preservation for growing in UV-B elevated climate.

57-63

Authors: Shipra Kalra, Kanav Midha, - Monika, Manpreet Kaur

Lysyl oxidase produces H2O2 i.e. ROS by acting on L-Lysyine. In the present study, ROS thus produced has been used by in vitro cytotoxicity assay on ovarian cancer cells thereby achieving 250 percent inhibition. Lysyl oxidase activity was determined spectrophotometrically by Dintrophenylhydrazine (DNPH) reagent. For isolation of lysyl oxidase produced by Trichodermaviride, acetone precipitation and ammonium sulphate precipitation was carried out. The effect of temperature on lysyl oxidase production was determined by incubating the media with 1.5 % inoculum at different temperature ranging from 10 , 20, 30,40,50,60,70 , 80 0C for 72 hr . Anti-tumor properties of Lysyl oxidase was checked using Rhodamine assay and NBT Assay. Comparative results for Acetone and Ammonium Sulphate precipitation showed that Enzyme activity(U/ml) of acetone precipitation is 124.6 and 144.3 IU/ml and Protein Content is 1.8 and 2.2 mg/ml with the specific activity 68.4IU/mg and 64.1IU/mg . The optimum enzyme production was found to be at pH 8 and optimum temperature for lysyl oxidase production was 500C with maximum enzyme activity of 0.38 (U/ml). 7th fraction contained highest enzyme activity so the retention time of lysyl oxidase was found to be 7 min. The protein content was also calculated by using Bradford method and it was highest in the 1st fraction and in 6th, 7th and 8th fractions. The specific activity has been improved from 75.1 IU/mg to 86.5 IU/mg. 10 units of lysyl oxidase inhibits 3x104 cells in each well to 82.5% and inhibition achieved at same cell count at 200 units which was 250% as observed with NBT and Rhodamine assay.

67-73

Authors: Dastigir Sayyad Mundaganur, Yasmin Dastigir Mundaganur, Ashokan K.V.

Keeping soil quality at par is one of the needed things for sustainable development and the existence of millions of living things in biosphere. Soil enzymes are used as soil quality indicators for quick response of changes for environmental stress, pollution and agricultural practices much more sooner (1–2 year) than other soil properties (organic matter); easy to measure (relatively simple procedure), having relations with plant productivity, soil quality parameters (organic matter, soil physical properties, microbial activity, and microbial biomass), and biogeochemical cycle; and being integrative. To assess the detrimental effect of the soil in grape cultivating field we selected four farming sites and various soil enzymes like protease. Urease, cellulose, chitinase, beta glocosidase, phosphatase, amylase, aryl sulphatase and dehydrogenase. As an additional support, we estimated the microbial population in all these fields and in all the stages of the cyclic process of the vine cultivation. Our study showed that the extensive use of the chemical pesticide badly affect the soil microorganism and which in turn badly affect the quality and quantity of the soil enzymes and subsequently the quality of the soil.

79-81

Authors: Rajani Shrestha, sheetal vaidya

External morphology of teleost brain is organized in such a way that it reflects the correlation between sensory adaptation and principal modes of activity very clearly. Channa gachua, Garra annandelei and Heteropneustes fossilis are among the 168 fresh water fishes recorded in Nepal. While examining the gut contents of these fishes, it was found that C. gachua fed primarily on insects, G. annandelei fed mainly on filamentous algae whereas H. fossilis fed both on plants and animals. Their brain morphology revealed that olfaction in C. gachua was stronger than in G. annandelei and H. fossilis. Similarly, optic sense was also more powerful in C. gachua than in G. annandelei and in H. fossilis. On the contrary, cerebellum of H. fossilis was more developed than that of G. annandelei and C. gachua. These morphological differences of brain could be undoubtedly correlated with the carnivorous, herbivorous and omnivorous feeding behaviour of C. gachua, G. annandelei and H. fossilis respectively. Moreover, in all three fishes, around 20% of the total gut content was made of mud and sand, and they all had inferior mouths, indicating that they were bottom feeders.

92-103

Authors: Md.Jahirul Islam, Kaniz Fatema, Pipasha Biswas

Zaire ebolavirus (EBOV) is one of the dangerous and a negative-stranded ssRNA virus. EBOV is a zoonotic pathogen that causes severe hemorrhagic fever in humans. Nowadays epidemic outbreak caused by EBOV is incurable with present technologies; thus figure out as a major health risk which needs enhanced surveillance. The study was conducted with seven proteins of Zaire ebola (EBOV) and gene sequences are available in NCBI database. The homology modeling was done by SWISS-MODEL, Phyre2 and HHpred. The obtained model was verified with structure validation programs such as PROCHECK, Verify3D and ERRAT. PROCHECK analysis of seven proteins showed that 85-96.6% of the residues are in the most favored region, the verify 3D value of 80-100% indicates that constructed model is good and ERRAT value of 87.442-100% indicates that overall good quality factor. In this study, we also reported phylogenic relationship, physico-chemical characteristics, secondary structure,3-D structure. Moreover, active sites were identified by CASTp suggests that these proteins can be utilized as a potential drug target. Furthermore, the initial findings were reinforced by the results from I-Mutant and mCSM as these tools predicted significant and functional instability of the mutated vp35 protein.

113-116

Authors: B.P. Yadav, D. N. Yadav, K.B. Koirala, K.R. Pandey, R.B. Thapa

Non-primed top quality seed of maize having cent-percent germination in lab resulted only 61% emergence in the field based on 4806 observations for non-primed seed while treated seed (priming cum incubation) recorded 82 % emergence in the field based on 4806 observations during first year. During second year experiment also non-primed top quality seed of hybrid maize having cent-percent germination in lab resulted 71.29% emergence while priming cum incubation treatment recorded 91.84% emergence. The result of both years clearly indicates the importance of seed treatment for maintaining the optimum plant population and there by insuring higher yield of winter hybrid maize.

130-132

Authors: Binoy S Vettical

The objective of the study was to evaluate the quality of semen retrieved from cauda epididymis of crossbred bulls in the tropics. Testes from one hundred and twenty six recently slaughtered cross bred bulls in the tropics were used for the analyais. The total and progressive motility percentage obtained for epididymal semen were 49.17±9.26 per cent and 27.5±9.11 per cent respectively. The mean concentration obtained for epididymal semen was 37,175x106 ±7612x106 per ml. The mean percentage of live and dead sperms was 84.5±8.02 per cent and 15.5±8.02 per cent respectively. The mean percentage of normal spermatozoa, spermatozoa with abnormal heads, abnormal tails, spermatozoa with a proximal protoplasmic droplet and distal protoplasmic droplet were 35.67±2.30, 3.17±1.58, 2.33±0.61, 11.67±4.01 and 47.17±3.17 per cent respectively.