029183   MANAGAMURI U , VIJAYALAKSHMI M, INDUPALLI M D , GANDURI V S R K, RAJULAPATI S B , PODA S (Acharya Nagarjuna Univ, Guntur-522 510, Email: profmvl08@gmail.com) : Improved bioactive metabolite production by Saccharopolyspora halotolerans vsm-2 using response surface methodology and unstructured kinetic modelling. Pharmacogn J 2018, 10(5), 833-40.

This study targets to optimize and analyse the interactive effects of process variables for improved bioactive metabolite production using RSM and unstructured kinetic modelling by S. halotolerans VSM 2. RSM was applied to optimize the interactive effects of five variables, viz., time of incubation, pH, temperature, concentration of maltose and meat extract on bioactive metabolite production and its effect against the five responses viz., S. flexneri, S. marcescens, P. vulgaris, P. aeruginosa and E. coli. Models of Logistic and Luedeking-Piret were used to simulate the cellular increase and bioactive metabolite production. RSM optimal conditions for the bioactive metabolite production recorded were incubation time (12days), pH (8), and temperature (250 C), concentrations of maltose and meat extract (1 % w/v) (each). The effect of the bioactive metabolite produced (zone of inhibition) against the responses were found to be 17 mm for S. flexneri, 17 mm for S. marcescens, 16 mm for P. vulgaris, 17 mm for P. aeruginosa and 18 mm for E coli. The data obtained from experimental values are in close agreement with the predicted values of RSM. Model adequacy was evaluated using ANOVA variance where the quadratic effect of p<0.0001 which imply the significance of the model. The unstructured-, mathematical- kinetic models provided a better approximation of profiles of S. halotolerans VSM 2 growth, optimized media utilization and bioactive metabolite production. Optimization of the independent variables for the production of the bioactive metabolite using RSM by S. halotolerans VSM 2 and its effect against the five responses were documented. The predicted values are in good agreement with the experimental values. Unstructured models provided a better approximation of kinetic profiles for bioactive metabolite production by S. halotolerans VSM 2.

6 illus, 5 tables, 34 ref

029189   MISHRA P, BHATT D K (Bundelkhand Univ, Jhansi - 284 128, Email: piyushmishrajhs@gmail.com) : Evaluation of nutritional and physico-chemical characteristics of dietary fibre enriched with carrot pomace pasta. Int J Pharm Sci Res 2018, 9(10), 4487-91.

Fibre rich pasta were prepared by substituting refined wheat flour with carrot pomace powder (CPP) at 5 %, 15 %, and 25 % and evaluated for its physical properties, chemical composition, textural properties and sensory characteristics. The fat content of the pasta showed no pronounced variation. The hardness of the pasta increased with increase in the level of incorporation of CPP in the flour blends. CPP improved the appearance of pasta by imparting it attractive colour; however the texture score decreased with the increase in the level of supplementation. Pasta with 15 % CPP was found to be most acceptable due to attractive appearance; and better taste and flavour.

1 illus, 7 tables, 12 ref

029168   HAQ I-U, MAHER A, GILL N P, FATIMA U, QADIR G, ALI K (Sindh Univ, Jamshoro - 76080, Pakistan, Email: rao.ikram@yahoo.com) : Growth of Bacillus subtilis and production of acetic acid with rotten potato: Used as substrate. Int J Pharm Sci Res 2018, 9(10), 4229-35.

The Bacillus subtilis is anaerobic gram-positive, dominant and nonpathogenic bacterial workhorses in industrial fermentations. In present work, B. subtilis LC4 cultured on a number of combinations of TY-medium with peels and peeled-off rotten potatoes to investigate its growth and level of secondary metabolites production including acetic acid. Cell culture was harvested after 42 and 62 h and maximum growth rate was observed in LB5 (peeled off potato extract) and LBo (TY-medium) cultures, while culture OD600 was low at 62h of all. It might be reduced with the shortage of nutrients in the medium that lead to killing of cells. Reducing sugars and total proteins were observed higher (p≤0.05) but reversed in both harvests to the decreasing rate of cell growth or multiplication rate of all cultures. Similar trend in production of acetic acid was also noted (p≤0.05) in all cultures. Increasing level of reducing sugars concentration among the culture indicates the cells are growing under nutritional stress. Meanwhile production of acetic acid along the higher concentrations of reducing sugars could be involved to initiate signals for sporogenesis in Bacillus subtilis cells. It could be a best adjustment of Bacillus cells for their survival in nutrient medium at stationary or decline growth phase. Both mixture of peels and starch extracts of rotten potato are recommended as the best productive and cheapest substrate for growth of Bacillus subtilis as well as may be recommended for microbial based extracellular proteins and acetic acid production.

3 illus, 1 table, 56 ref

029163   GALANDE S, KHAURSADE P, PRAKASHAM R S, KISHORE P B K (CSIR-Indian Institute of Chemical Technology, Hyderabad - 500 007, Email: prakasam@iict.res.in) : In-silico development of efficient l-asparaginase enzyme for acute lymphoblastic leukaemia therapy. Int J Pharm Sci Res 2018, 9(10), 4177-86.

L-asparaginase has been accepted clinically as an anti-tumour agent for the effective treatment of acute lymphoblastic leukaemia and lymphosarcoma. This enzyme also possesses L-glutaminase activity and causes immunological problems. Hence, efforts have been made to develop mutants with lower or no glutaminase activity. In the present study, a homology model of L-asparaginase obtained from Pectobacterium carotovorum was docked and compared with its mutants for activities, analysed for molecular dynamics and structural stability. A total of five in-silico mutants were developed using single amino acid mutagenesis and evaluated for ligand binding for both L-asparagine and L-glutamine. One of the mutants, Y306L, showed -5.89 and -5.04 while the wild L-asparaginase revealed -5.50 and -5.12 binding energies for L-asparagine and L-glutamine, respectively. Further, substrate-enzyme interaction analysis indicated that the wild L-asparaginase showed five interactions for L-asparagine as well as for L-glutamine, whereas, the mutant, Y306L depicted eight and three interactions for L-asparagine and L-glutamine. Molecular dynamics analysis denoted that mutant protein is more stable in RMSD, RMSF and radius of gyration to that of wild-type protein. Higher binding affinity was noticed with Lasparagine and lower with L-glutamine along with higher interactions with Lasparagine and lower with L-glutamine in mutant Y306L, compared to wild Lasparaginase. This suggests that this could be a possible potential candidate for the treatment of Acute Lymphoblastic Leukaemia (ALL) with less induced side effects.

12 illus, 2 tables, 16 ref

029192   PURNAMAWATI F, SANTOSO S, AULANNI’AM (Brawijaya Malang Univ, Brawijaya Malang, Indonesia, Email: d.mateen7181@gmail.com) : Detection of B. abortus S19 on uterine rat (Rattus norvegicus) with immunohistochemical method. Int J Pharm Sci Res 2018, 9(9), 4021-8.

Brucellosis in humans is the most common zoonotic infection in the world. Brucellosis can be prevented by vaccination but no effective vaccines are known for prevention of human brucellosis and minimal impact of animal brucellosis. Outer membrane proteins (OMPs) of Brucella sp. have been the focus of vaccine development and the diagnosis of brucellosis. Methods: The pathogenesis study of brucellosis used rat by intraperitoneal injection. Profile OMP detected by SDS-PAGE. The antibodies took from rabbit and the immune response did with western blotting, dot blot and immuno histochemistry. Results: B. abortus S19 cell to the rat show the presence of inflammatory cells and bleeding in the uterine rat. The 36 kDa OMP could isolated from B. abortus S19. Rabbit antibody to 36 kDa OMP can detect B. abortus of this histological of the uterus. OMP 36 kDa B. abortus S19 is antigenic characterized by its ability to stimulate antibodies in the host's body (Oryctolagus cuniculus). Conclusion: B. abortus S19 can be detected in histopathology of rats uterus shown by interaction of antigen and antibody by using immunohistochemical method.

4 illus, 26 ref

029160   DEV S S, VINEETHA K U, FRANCIS S (Biotechnology Dep, St. Peter’s Coll, Kochi - 682 311, Email: sonaaniyan@gmail.com) : Green synthesis of silver nanoparticles using leaf extract of Ayapana triplinervis and its antibacterial activity. Int J Pharm Sci Res 2018, 9(9), 3897-902.

Green synthesis of nanoparticle using plant extracts is an efficient alternative for the conventional process of preparation of nanoparticles by chemical and physical methods which leads to non-ecofriendly by-products.The main aim of the project was to produce silver nanoparticles (AgNPs) using leaf extracts of Ayapana triplinervis (Eupatorium triplinerve) a medicinally important herb and to ascertain its antibacterial properties. AgNPs were synthesized by adding 1 ml leaf extract of A. triplinervis to 10ml of 3mM silver nitrate followed by incubation at 60 °C. The AgNPs produced were further characterized by ultraviolet-visible (UV-Vis) spectroscopy, Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM) analysis. UV-Vis spectroscopy of prepared silver colloidal solution showed absorption maximum between 460 - 490 nm due to the excitation of surface plasmon resonance (SPR) of the AgNPs. FTIR spectra absorption peaks at 3431, 2065, 1635 and 530 cm⁻¹ revealed the availability of functional groups which may be involved in the AgNPs synthesis. The SEM analysis showed that AgNPs were spherical or ellipsoidal in shape.The antibacterial activity against common pathogens was evaluated by well diffusion method. Salmonella, Pseudomonas and Bacillus gave positive results. The AgNPs produced in this study by green synthesis methodology from leaf extract of A. triplinervis needs to be further evaluated and could find greater application in pharmaceutical industry.

5 illus, 35 ref

030580   MAHAPATRA K D, SAHOO L, SAHA J N, MURMU K, RASAL A, NANDANPAWAR P, DAS P, PATNAIK M (Fish Genetics and Biotechnology Div, ICAR-Central Institute of Freshwater Aquaculture, Bhubaneswar - 751 002, Email: kdmahapatra@yahoo.co.in) : Establishment of base population for selective breeding of catla (Catla catla) depending on phenotypic and microsatellite marker information. J Genet 2018, 97(5), 1327–37.

The phenotypic and microsatellite marker information of nine strains of catla (Catla catla) for growth trait was used to infer relationship within and between strains. This information helped in optimizing the proportion of individuals to be used from each strain while creating a base population for selective breeding. For this purpose, nine strains were collected from different sources and places of India namely West Bengal, Bihar, Odisha, Andhra Pradesh and Uttar Pradesh. Two riverine sources i.e. Ganga and Subarnarekha were also represented among the nine strains collected for base population. They were brought to Indian Council of Agricultural Research-Central Institute of Freshwater Aquaculture (ICAR-CIFA) at fry stage and reared separately till fingerlings. After passive integrated transponder tagging fingerlings were stocked in three communal ponds for one year culture. Live body weights were then measured and least square means were obtained after pond effect correction. A wide range of variation was observed among and between strains. Microsatellite markers were used to estimate genetic differences of different strains of catla using pair wise F_ST estimates. Overall multi locus F_ST, including all loci was estimated to be 0.4137 (P < 0.05), indicating genetic heterogeneity among them. Analysis of molecular variance revealed that, 58.63 % of variation was due to within individual variation, 3.45 % of variation was due to among individuals within strain and 37.92 % was due to among strain variations. Both phenotypic as well as microsatellite data will be used to form a base population of catla with individuals from the stock having broad genetic variation for selective breeding programme.

5 illus, 6 tables, 44 ref

030584   PAVANI M, SUNDARAM R M, RAMESHA M S, KISHOR P B K, KEMPARAJU K B (Biotechnology Dep, Indian Institute of Rice Research, Hyderabad - 500 030, Email: KB.Kemparaju@icar.gov.in) : Prediction of heterosis in rice based on divergence of morphological and molecular markers. J Genet 2018, 97(5), 1263–79.

Identifying the best performing hybrid without a field test was essential to save resources and time. In this study, the genetic divergence was estimated using morphological and expressed sequence tag (EST)-derived simple sequence repeats (SSR) markers. Cluster analysis showed that APMS6A and RPHR 1005 belong to groups I and II, respectively, and the hybrid combination recorded the highest mean grain yield of 32.25 g among generated 40 F₁s with standard heterosis of 8.4 % over hybrid check, KRH2. The coefficient of marker polymorphism (CMP) value was calculated based on EST-SSR markers; it ranged from 0.40 to 0.80, and a higher CMP value of 0.80 was obtained for the parental combination APMS6A × RPHR1005. We predicted heterosis for 40 F₁s based on correlation between CMP and standard heterosis in different traits with standard varietal and hybrid checks indicating positive correlation and significant value for grain yield per plant (r = 0.58**), productivity per day (r = 0.54**), productive tillers (r = 0.34*) and panicle weight (r = 0.42**). This study revealed that the relationship of molecular marker heterozygosity, along with the combining ability, high mean value of different traits, grouping of parental lines based on morphological and molecular characterization is helpful to identify heterotic patterns in rice.

4 illus, 11 tables, 31 ref

030590   SENTHILKUMAR P, DAWN S, SAIPRIYA C, SAMROT A V (Biotechnology Dep, Sathyabama Institute of Science and Technology, Chennai, Email: antonysamrot@gmail.com) : Synthesis of polyhydroxybutyrate nanoparticles using surfactant (SPAN20) for hydrophobic drug delivery. Rasayan J Chem 2018, 11(4), 1686-95.

Biopolymers like polyhydroxybutyrate (PHB) are biocompatible and biodegradable, it forms nanoparticle when appropriate solvent systems are employed. Thus, they fulfill the major requirement as a drug carrier. In this study, surfactant (Span20) influenced PHB nanoparticles were prepared by nanoprecipitation method with different solvent systems i.e. Chloroform:DMSO (CD), Chloroform: Water (CW), Ethylacetate: DMSO (ED) and Ethylacetate:Water (EW). The nanoparticles were also loaded with a hydrophobic drug - curcumin. Further, the produced nanoparticles were characterized and utilized for in-vitro drug release studies. The nanoparticles were below 300 nm in size and these nanoparticles were found to release the drug for longer duration i.e. more than 110 minutes. From the controlled release studies carried out against Bacillus subtilis, it was found that the nanoparticles released the encapsulated drug (Curcumin) efficiently, when acetic acid was used as the solvent system.

5 illus, 1 table, 49 ref

030602   SURYAPRABHA M, RAO N R (Biotechnology Dep, Chalapathi Institute of Pharmaceutical Sciences, Guntur - 522 034, Email: surya_pharm@yahoo.com) : Pharmacological evaluation of polyherbal extracts of Amorphophallus paeoniifolius, Citrus sinensis and Plumbago zeylantia for anti fungal activity. Int J Pharm Biol Sci 2018, 8(4), 1261-4.

Fungal infections may take the form of superficial infections involving the skin or mucous membranes or systemic involving various internal organs. Deep infections are systemically treated with drug therapy. Among the fungi that cause systemic infections are Aspergillus spp., Nocardia asteroides , Histoplasma capsulatum and Coccidiodes immitis. Among ancient civilizations, India has been known to be rich repository of medicinal plants. Recently, WHO estimated that 80 percent of people worldwide rely on herbal medicines for some aspect of their primary health care needs. Treatment with medicinal plants is considered very safe and there is no or minimal side effects. Medicinal plants are considered as rich sources of ingredients which can be used in drug development. The Ayurvedic literature Sarangdhar Samhita’ highlighted the concept of polyherbalism to achieve greater therapeutic efficacy. The active phytochemical constituents of individual plants are insufficient to achieve the desirable therapeutic effects. When combining the multiple herbs in a particular ratio, it will give a better therapeutic effect and reduce the toxicity. This research mainly focuses on important of the polyherbalism and its clinical significance.

1 illus, 1 table, 10 ref

030571   KALA K, GOPINATH L R, ABINAYA K, BHUVANESWARI R, ARCHAYA S (Biotechnology Dep, Vivekanandha Coll of Arts and Sciences for Women, Namakkal, Tamilnadu, Email: lrgopinathnamakal@gmail.com) : Identification of biologically active compounds from Withania somnifera leaf extract its antimicrobial activity and anticancer property. Int J Pharm Biol Sci 2018, 8(4), 1237-44.

Medicinal plants were identified by traditional communities in ancient times through their constant interaction with them. Modern medicines which were based on non-organic in origin lead to numerous complications interms of side effects and resistant genes. Hence more and more attention is paid to the medicines based on organic origin. Among numerous plants that have been used by traditional communities Withania somnifera one plant which was used for wide range of therapeutic properties. However, in the present study an attempt have been made to identify effective solvent and biologically active compounds for antimicrobial and anticancer property. Among the three-solvent water, methanol and ethanol. Methanol was found to be effective in identifying secondary metabolites and also quantity of secondary metabolites was higher when compared to other solvents. Phenols were found to be higher in the W. somnifera leaves followed by flavanoids and alkaloids indicating their therapeutic potential. GCMS results of methanol crude extract of W. somnifera showed 24 different compounds with alkanes, alkenes, alcohols, esters, etc. which are known for anticancer and antimicrobial activity. Antimicrobial activity among the tested organisms K. pneumonia showed the highest followed by P. mirabilis and P. aeruginosa comparable with the standard antibiotics. Antioxidant capacity of W. somnifera quantified through DPPH and ABTS assay. The DPPH scavenging activity increased up to 86.71 % with 400 μg/ml and ABTS reached up to 92.91 with 400 μg/ml. The methanol crude extract of W. somnifera showed least IC50 with HeLa 194.11 μg/ml followed by MCF 210.78 μg/ml HepG2 217.98 μg/ml cell lines.

8 illus, 3 tables, 15 ref

030561   CHOUDHARY S, SHARMA N (Bioscience and Biotechnology Dep, Banasthali Vidyapith, Vidyapith - 304 022, Rajasthan, Email: sangeeta25@gmail.com) : Copper uptake and transport across physiological barriers. Int J Pharm Biol Sci 2018, 8(4), 1231-6.

Copper is an essential multi-functional, micro-nutrient which plays a crucial role in human physiology. This element is found in various proteins and enzymes having significant function in metabolism, development and maintenance of different organs and systems. Diverse organisms, from bacteria to mammals have developed elegant mechanisms of copper uptake, transport and storage. Balance of this micronutrient is maintained by complex homeostatic processes leading to constant and sufficient supply of it and avoids any excess accumulation. Regulation of cellular copper (Cu) homeostasis involves CTR1protein, responsible for Cu uptake across the plasma membrane, the Cu chaperones and Cu transporting ATPase’s (Cu-ATPase’s), i.e., ATP7A and ATP7B.This manuscript presents a review on the copper transport across the human physiological barriers, delineating its role in metabolism and the multiple steps of metal assimilation. Our attention is focused to copper interaction and trafficking in the gut and to other systems such as blood, brain and placenta.

2 illus, 30 ref

030570   JHARIYA N, PATHAK S (Biotechnology Dep, Mata Gujri Mahila Mahavidyalaya, Jabalpur - 482 001, Email: sonal_sareen2006@yahoo.com) : Studies on heavy metal tolerance of fungal isolates from soil samples of Jabalpur region. Int J Pharm Biol Sci 2018, 8(4), 1167-76.

The presence of heavy metals in the environment has been a subject of great concern due to their toxicity and non-biodegradable nature. The fungal biomass can act as bio-sorbent for the removal of heavy metals and radio nuclides from polluted waste materials. In the present study, the heavy metal tolerant fungi were recovered from soil samples of iron workshop site and waste dumping sites of Jabalpur region. A total of 26 isolates were obtained in the present study and were then assessed for their growth in presence of heavy metals like Cr, Co, Hg, Zn and Mn. Among the fungal isolates, Aspergillus species Penicillium species and Cephalosporium species showed tolerance with the heavy metals used. The determination of heavy metal tolerance of Aspergillus sp, Penicillum sp and Cephalosporium sp was done at concentration range (100 mM- 500 mM) for Cr, Cu, Hg, Zn and Mn respectively. It was found that the tolerance limit of Aspergillus species to be 19 mm in case of 120 mM Cr, Penicillium species showed tolerance limit of 7 mm with 160 mM Cu and Cephalosporium species showed 21mm with 180 mM Cr. The results suggest potential of soil mycotic flora of Jabalpur region for the removal of heavy metal from different sites.

4 illus, 4 tables, 15 ref

030603   SURYAWANSHI S S, RANE M R, KSHIRSAGAR P R, KAMBLE P P, JADHAV J P (Biotechnology Dep, Shivaji Univ, Maharashtra - 416 004, Email: profjpjadhav@gmail.com) : Antioxidant, antimicrobial activity with mineral composition and LC-MS based phytochemical evaluation of some Mucuna species from India. Int J Pharm Bio Sci 2018, 8(4), 1061-70.

The study was designed to demonstrate the phytochemical composition and to evaluate minerals, antioxidant and antibacterial activities of different Mucuna species. Liquid Column Mass Spectrometry (LC-MS) analysis was performed to identifies the phytoconstituents like phenolic, flavonoids, alkaloids, vitamins, amino acids, anti-cancer agent, antiretroviral agents, antidiabetic agents, anti-inflammatory and anti-Parkinson’s drug. Minerals essential for healthy functioning of the human body were resoluted from these species using atomic absorption spectroscopy. The antioxidant activity was evaluated by using the DPPH and ABTS assay. The antimicrobial activities of various Mucuna extracts were performed against some micro-organisms (Gram positive and Gram-negative micro-organism) using agar well diffusion method. The results of the present study revealed variation between some different Mucuna species. The results of LCMS phytochemical profiling indicate flavonoids and related polyphenols responsible for various pharmacological activities were the major compounds in the Mucuna seeds. Estimated mineral content in various Mucuna species shows the importance of these new wild varieties (Mucuna bracteata and Mucuna imbricata). Thus, serve as a great platform for the food industry due to their macro and micronutrients contents in significant amount. The strong antioxidant and antimicrobial activity present in the wild Mucuna imbricata seeds may be good alternative to the communally used of Mucuna species. They can also be used as a potential source of food and anti-Parkinson’s drug in the pharmaceutical industry. Thus, the present article puts focus on the possible use of two wild Mucuna species as a substitute for Mucuna pruriens var utilis and herbal remedy against neurodegenerative disorders.

3 illus, 1 table, 49 ref

030588   RANJITH SANTHOSH KUMAR D S, SENTHIL KUMAR P, SURENDRAN L, SUDHAGAR B, VINODHINI J (Biotechnology Dep, Kongunadu Arts and Science, Coimbatore - 29, Email: senthilkumarp1185@gmail.com) : Isolation, characterization, and differentiation of mesenchymal stem cell from rat bone marrow in vitro. Int J Pharm Biol Sci 2018, 8(4), 1014-23.

Bone marrow mesenchymal cells have been identified as a source of pluripotent stem cells with varying degrees of plasticity in humans. MSCs have the potential to differentiate into different cell types such as osteoblast, chondroblast, adipoblast. This research describes the pioneering experiment of isolation and differentiation rat bone marrow stromal cells into the osteoblast and adipoblast cells lineages. Bone marrow stromal cells were isolated from long bones of rat, based on the adherent properties of the MSC. The stromal cells obtained by direct plastic adherence were characterized by fluorescent activating cell sorting (FACS) for established hematopoietic and non-hematopoietic markers. Under the determinate effect of culture conditions, MSCs were differentiate done osteogenic and adipogenic cell lines detected by Alizarin red and oil-red O staining invitro. Bone marrow samples from rats yielded 4-5 million bone marrow mononuclear cells/ml per femur. The MSCs culture is being investigated with microscopic observations. They form elongated or spindle-shaped fibroblast-like cells. Cell sorting by the surface marker expression is used in the current studies for confirmation of rat MSCs in the culture. Moreover, the cells differentiated toward osteoblasts and adipocytes were characterized morphologically by Alizarin Red staining and Oil Red O staining. This research consigns new ideas to develop noble markers and play a vital role in tissue engineering.

8 illus, 53 ref

030569   JEYA K R, VEERAPAGU M, VETRIKODI N, PRIYA R, LATHA G (Biotechnology Dep, Bharathidasan Univ Constituent Model Coll, Veppur - 621 717, Email: krjaya5@gmail.com) : Screening and production of alkaline protease from halophilic bacteria isolated from solar saltern. Int J Pharm Biol Sci 2018, 8(4), 948-53.

Halophiles are predominant organisms which are present in the hypersaline environment saltern ponds. They produce a variety of industrially important bioactive compounds including protease enzymes. Soil samples were collected from solar saltern of Vedaranyam, Tamilnadu. A total of 12 halophilic bacteria HPB1 –HPB12 isolated were screened for proteolytic activity by plate assay showed that seven isolates exhibited proteolytic activity by zone formation around the colonies. Isolate HPB2 produced high amount protease enzyme 35.4 U/ml than the other seven isolates were identified as Bacillus megaterium. The effect of incubation time, medium pH, temperature, NaCl concentration and carbon source source for the protease production was investigated. Protease enzyme production by Bacillus sp was maximum at 40 hrs of incubation period 86.5 U/ml, pH 8 (107.2 Uml^-1), temperature 40 ^oC (98.3 Uml^-1), Nacl 2M (102.8 Uml^-1). and glucose (124.5 Uml^-1) as carbon source.

5 illus, 1 table, 34 ref

030566   GOPINATH L R, RAMYA BHARATHI V, ARCHAYA S, PREMLATHA K (Biotechnology Dep, Vivekanandha Coll of Arts and Sciences for Women, Namakkal, Tamilnadu, Email: lrgopinathnamakkal@gmail.com) : Identification of biologically active compounds from Adhatoda vasica leaf extract its antimicrobial activity and anticancer property. Int J Pharm Biol Sci 2018, 8(4), 927-34.

Adhatoda vasica is one such plant which is used widely for a variety of purposes which forms part of many other traditional herbal medicines. Biologically active compounds present in the plants are responsible for the such curative activity. These compounds could be identified by dissolving them in appropriate solvents. In the present study water, methanol and ethanol were used as solvents. Most of the secondary compounds were found in all the solvents but methanol gave positive result more than the other. Quantification of important solvents showed that high tannin content 65.61 μg/ml followed by saponins 19.09 μg/ml and alkaloids 12.87 μg/ml with methanol crude extract of A. vasica. GCMS analysis of methanol crude extract of A. vasica showed 21 compounds of alcohols, steroids, ester, etc. mostly having antimicrobial and antioxidant property. Antimicrobial property with well diffusion method showed effective antimicrobial activity for B. subtilis 12.17 mm followed by V. cholera 11.83 mm and K. pneumonia 11.50 mm. Antioxidant activity was performed with DPPH assay and ABTS assay with DPPH assay 81.81 % antioxidant activity was recorded at 160 μg/ml and 94.84 % antioxidant activity with 160 μg/ml using methanol crude extract of A. vasica. Anticancer property was estimated through cytotoxicity study with cell lines showed least IC₅₀ with HeLa 88.24 μg/ml followed by MCF 92.80 μg/ml HepG2 111.08 μg/ml cell lines using methanol crude extract of A. vasica.

7 illus, 3 tables, 16 ref

030592   SHARMA R, ZIMIK M, ARUMUGAM N (Biotechnology Dep, Pondicherry Univ, Pondicherry - 605 014, Email: arumugan.dbt@pondiuni.edu.in) : Isolation and GCMS characterization of certain non-polar compounds from Spilanthes ciliata. Int J Pharm Biol Sci 2018, 8(4), 889-903.

Spilanthes ciliata, commonly known as toothache plant, is a traditional medicinal plant. In the present study an attempt was made to analyze the oil obtained after solvent extraction of flowers of the plant. GCMS of the extract revealed presence of 15 different compounds in the oil. 11-tridecene-1-ol (18.72 %), 2R-acetoxymethyl-1,3,3-trimethyl-4T-(3-methyl-2-buten-1-yl)-1T-cyclohexanol (12.66 %), Pentadecanoic acid (8.36 %,), 1-hexacosanol (8.32 %,), Nonadecane (7.35 %) and Hexatriacontane (7.014 %) were found to be the major constituents and the remaining were in trace amounts. Most of the compounds identified are found associated with various biological activities. The study presents a large-scale isolation of major compounds for their characterization and medicinal application.

3 illus, 1 table, 32 ref

030568   JENAT P J, SURESH S N (Biotechnology Dep, Sree Narayana Guru Coll, Coimbatore - 641 105, Email: drsnsuresh78@gmail.com) : Antioxidant activity and antibacterial activity of Adiantum lunulatum Burm.f.. Int J Pharm Biol Sci 2018, 8(4), 779-82.

Medicinal value of Adiantum lunulatum due to the presence of the phytochemicals and this phytochemicals have major role in the antioxidant and antibacterial activity. DPPH assay and antibacterial activity analysis help to identify the medicinal potential of this fern. Agar well diffusion method and minimum inhibitory concentration analysis (0.2 mg/ml, 0.4 mg/ml, 0.6 mg/ml, 0.8 mg/ml and 1 mg/ml) carried out against bacterial strains such as Staphylococcus aureus, Klebsiella pneumonia, Proteus vulgaris and Escherichia coli. DPPH assay help to identify the free radical scavenging activity of plant extract. The result indicates that increase concentration of extract has direct influence corresponding to the percentage of free radical scavenging activity. Bactericidal activity analysis shows more bacterial growth inhibition at 0.8 mg/ml and 1 mg/ml against most of bacterial strains. Adiantum lunulatum show increased effectiveness against various diseases and need further research study.

1 illus, 3 tables, 15 ref

030582   MASHHOOR K, RAMESH N, LAZAR K V, SHANAS S (Biotechnology Dep, E.M.E.A. Coll of Arts and Science, Malappuram - 673 638, Kerala, Email: mashhoork@gmail.com) : Phylogenetic status of rice dark headed stemborer, Chilo polychrysus. Int J Pharm Biol Sci 2018, 8(4), 768-72.

Generally, stemborers were considerd as the most serious pest of the paddy. The stemborer larvae enter to the tiller and feed inside, which leads to characteristics damage symptoms of ‘dead hearts’ or ‘white ears. Five pyralid rice stem borer species were reported from India and the phylogenetic relationship of them was not yet studied in detail. In this study we have determined the phylogenetic status of rice dark headed stemborer Chilo polychrysus using Cytochrome oxidase subunit I (COI) partial coding sequence. The COI sequence of C. polychrysus showed 9 %, 10 and 13 % divergence with stem-borers viz. C. suppressalis, Scirpophaga excerptalis and Scirpophaga innotata respectively. Interestingly it showed only 6-8 % divergence with many species of different genus of Lepidotera family. Phylogenetically Lampides boeticus, Maculinea ario, Oleria olerioides, Oleria fumata, Greta andromica and Pseudoscada florula are the nearest relative of C. polychrysus. In the phylogeny tree the C. polychrysus and C. suppressalis were not aligned in a same clad. Therefore, further studies are needed to determine the taxonomic position of C. polychrysus.

2 illus, 1 table, 11 ref

030576   KUMAR D, GAJBHIYE A, NAGDA V, ARORA A (Biotechnology Dep, Sir Padampat Singahania Univ, Udaipur - 313 601, Email: dinesh.chhatwani@gmail.com) : Characterization of root extracts of Withania somnifera and effect of zinc sulphate and silver nitrate on antibacterial activity against certain human pathogenic bacteria. Int J Pharm Biol Sci 2018, 8(4), 682-9.

There is a serious need for the expansion of new antibacterial agents from natural sources due to emergence of new infections and increase in bacterial drug resistance that have been developed over a period of time in bacteria. The present study was carried out to evaluate the effect of zinc sulphate solution on antibacterial activity of crude methanolic and aqueous extract of W. somnifera root and its phytochemical characterisation. Agar well diffusion assay, as well as micro broth dilution assays, were used for determination of antibacterial activity. The assay was performed in triplicate and tetracycline was used as the control. The methanolic and aqueous extracts of W. somnifera root was prepared by percolation method. Pathological isolates Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae were obtained from the Department of Microbiology, RNT Medical College & Hospital Udaipur (Raj.). Agar well diffusion method for antimicrobial susceptibility testing was performed. The antibacterial activities were assessed by the presence or absence of inhibition zones after incubating the plates at 37°C for 24 hours. Minimum inhibitory concentrations (MIC) for the selected pathogens were determined by broth micro dilution method. Among aqueous and methanolic extracts, methanolic extract was found to possess a more potent inhibitory activity with a maximum zone of inhibition of S. aureus in antibacterial susceptibility test. MIC value of the extract for S. aureus was found to be 3.4 ±0.003 mg/ml. Among all the combinations, maximum antibacterial activity was found with methanolic extract in combination with of zinc sulphate producing a zone of inhibition of 19.00 +0.33 mm. The antibacterial effect was similar in all combinations of aqueous extract and metal salts. Both aqueous and methanolic root extracts of W. somnifera found to possess antibacterial activity against E. coli and S.aureus. The extracts were ineffective against K. pneumoniae. The metal salts such as zinc sulphate enhances the antimicrobial potential of the root extracts. The methanol extract of W. somnifera roots possess the ability to control pathogenic bacterial growth and may be further analysed for medicinal value in ethno medicine.

2 illus, 5 tables, 37 ref

030573   KARMAKAR M, RAY R R (Biotechnology Dep, Maulana Abul Kalam Azad Univ of Technology, Kolkata - 700064, West Bengal, Email: raypumicro@gmail.com) : Fungal strain guided bottom-up synthesis of silver nanoparticles and elucidation of its anti-bacterial activity. Int J Pharm Biol Sci 2018, 8(4), 511-7.

Green synthesis of silver nanoparticles (AgNPs) were accomplished using biological source of fungal strain Rhizopus oryzae PR7. UV-Vis spectroscopy measurement of prepared AgNPs revealed a characteristic peak at ~420 nm. FT-IR spectroscopy indicated an inverse peak of ~330 cm^-1, 1450 cm^-1. The effect of enzyme concentration over the relative production of AgNPs has been also studied. The rate kinetics for the production of AgNPs showed that maximum conversion to its nano form was achieved within 60 mins. Anti-bacterial studies of prepared AgNPs performed over various Gram positive and Gram-negative target microorganisms revealed the effectiveness of Nano silver particles as antibacterial agents.

6 illus, 1 table, 15 ref

030604   VALLI S A, GOWRIE S U (Plant Biology and Plant Biotechnology Dep, Madras Univ, Chennai - 600 008, Email: umasezhian@gmail.com) : Plant-derived food products as dietary supplements for sustainable utilization- an in vitro and in silico approach. Int J Pharm Biol Sci 2018, 8(4), 416-25.

Sustainable utilization is the use of components of biological diversity in such a way that does not lead to its long-term decline, thereby maintaining its potential to meet the needs and aspirations of present and future generations. Investigation studies related to dietary supplements from natural edible food products have been one of the significant research interests in recent years. Using several in vitro and in silico methodologies, a study was carried out in leguminous sprouts to analyze the nutrients for its recommendation as dietary supplements for all age groups. The study was carried out with two different samples- horse gram sprouts (Macrotyloma uniflorum (Lam.) Verdc.) and mixed sprouts of combination (Cicer arietinum L. (Chick pea), Macrotyloma uniflorum (Lam.) Verdc. (horse gram) and Vigna radiata (L.) R. Wilczek (Green gram). Screening the phytoconstituents by qualitative phytochemical tests and quantification of the primary and secondary constituents were carried out in methanol and aqueous extracts of fresh and dried sprouts. Phytochemical characterization was carried out through FTIR and GC-MS analysis which proved the presence of essential phytoconstituents like terpenoids, fatty acids, proteins, carbohydrates and vitamins. Antibacterial activity of both the samples were studied against human pathogens namely Staphylococcus aureus, Escherichia coli, Salmonella typhi, Klebsiella pneumoniae and Shigella flexneri. Maximum zone of inhibition were shown by Shigella flexneri in horse gram and in mixed sprouts. Further, bioactive compounds obtained from GC-MS analysis were screened for anti-inflammatory activity and the potent bioactive compound- its structure and functions were analyzed through in silico studies using different bioinformatics tools and softwares which indicated the effective physico-chemical properties of the bioactive compounds which might be responsible for the anti-inflammatory (anti-ulcer) property. Thus, the horse gram sprouts and mixed sprouts enriched with the phytoconstituents can be recommended as a natural source of dietary supplements which can be one of the most economical and easily available source for consumption leading to sustainable utilization.

11 illus, 1 table, 22 ref

030597   SUBBURAYALU S, ASHA K R T, PALAVESAM A (Biotechnology Dep, Manonmaniam Sundaranar Univ, KR Nagar, Kovilpatti, Email: krplacement97@yahoo.in) : Evaluation of hepatoprotective effect of Indigofera tirunelvelica Sanjappa against CCL₄ induced hepatotoxicity in rats. Int J Pharm Bio Sci 2018, 8(4), 384-93.

The present investigations were undertaken to evaluate the hepatoprotective of the ethanolic extract of Indigofera tirunelvelica Sanjappa (Et-It) against CCl₄-induced hepatotoxicity in rats. Hepatoprotective effect of ethanolic extract of Indigofera tirunelvelica (EtL-It) was determined by using carbon tetrachloride (CCl₄) intoxication of rats as experimental models. The range of liver impairment and effect of the extract of medicinal plant was evaluated by assorted biochemical markers like aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine aminotransferase (ALT), total protein (TP) and total billrubin (TB) in blood serum and concentration of Lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), glutathione-s-transferase (GST) and glutathione peroxidase (GPx), in liver were determined. Histopathological changes in the liver of different groups were also studied. The administration of Et-It at dose levels of 100, 200 and 400 mg/kg/b.w., orally had decreased the rise of ALT, AST, ALP, TB and TBRAS levels and the effects were comparable to standard drug (Silymarin 20 mg/kg/b. w,) the GSH, SOD, CAT, GPx, GST and TP levels were significantly increased in the animals received EtL-It. The histopathological studies show decreased necrosis and hepatocellular degeneration when compared to the CCl₄ intoxicated liver. This study demonstrates that the hepatoprotective of Indigofera tirunelvelica therefore scientifically supports the use of this plant in traditional medicine for treatment of liver disorders.

6 illus, 29 ref

030567   HITHASHREE , KAVITHA, CHANDRASEKHAR N, GOVINDAPPA M, RAMACHANDRA Y L, CHANDRAPPA C P (Biotechnology Dep, Shridevi Institute of Engineering and Technology, Karnataka, Email: chandrappacp@gmail.com) : Synthesis of gold nanoparticles by the flower extracts of Tabebuia argentiea and their antioxidant activity. Int J Pharm Biol Sci 2018, 8(4), 379-83.

Biosynthesis of nanoparticles by plant extracts is currently under exploitation. Plant extracts are very cost effective and eco-friendly and thus can be an economic and efficient alternative for the large-scale synthesis of nanoparticles. The current study revealed that the aqueous flower extracts of Tabebuia argentiea were used and compared for their extracellular synthesis of gold nano-particles. Stable gold nanaoparticles were formed by treating aqueous solution of AuCl₃ with the plant flower extracts. The formed gold nano-particles were characterized by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX). The flower extracts act as reducing as well as encapsulating agent for the gold nanoparticles. The SEM revealed the formation of spherical gold nanoparticles with the average particles size of 56 nm. Antioxidant activity of gold nanoparticles was carried out and found to be more significant antioxidants.

4 illus, 1 table, 12 ref

030599   SUGANYA K, RAJESH SINGH J, GANESH P, SATNAMI P, ANTONY S (Biotechnology Dep, Rajah Serfoji Government Arts Coll, Thanjavur, Email: rajgenes@gmail.com) : The studies on understanding of midgut bacterial community of Aedes aegypti collected from Tamilnadu, India- A metagenomic approach. Int J Pharm Biol Sci 2018, 8(4), 360-8.

Metagenomics as a new field of research has been developed over the past decade to elucidate the genomes of the non-cultured microbes with the goal to better understand global microbial ecology on the one side, and on the other side it has been driven by the increasing biotechnological demands for novel enzymes and biomolecules. Ingestion of blood meal by female mosquitos triggers a series of physiological processes in the midgut and also exposes them to infection by these pathogens. The bacteria normally harbored in the mosquito’s midgut are known to influence physiology and can also alter the response to various pathogens. Here, we studied the bacterial diversity found in midgut part of Aedes aegypti to understand the host and microbe interaction. The adult Aedes aegypti mosquitos were collected from Chidambaram and around area using ovitrops and midgut part was extracted moreover the DNA templates were isolated and amplified by PCR. The DNA amplicons were sequenced by Illumina MiSeq gene sequence. The total 179,310 reads were classified in to 209, the bacterial genera of Aeromonas, Klebsella, Acinetobacter, Staphylococcus, Bacillus and Pseudomonas predominantly found to be high when compared to the other bacterial genera. The present data strongly encourage further investigations to verify the potential role of the detected bacteria in mosquito for the transmission several vectoral diseases.

4 illus, 2 tables, 36 ref

030565   GEETANJALI G, JAIN P, PUNDIR R K (Biotechnology Dep, Kurukshetra Univ, Kurukshetra, Email: drpranayjain@gmail.com) : Evaluation of antimicrobial potential of Aspergillus ibericus isolated from rhizospheric soil of Ficus religiosa. Int J Pharm Biol Sci 2018, 8(4), 320-31.

Inappropriate and irrational use of antimicrobial medicines has prompted the resistant microbes to emerge, spread and persist. Antimicrobial resistance may be intrinsic or acquired as it can develop through the spontaneous mutation of existing genes or through the transfer of genes from other resistant species or strains. Therefore, antimicrobial resistance has resulted research and development for the discovery of new antibiotics against pathogens at all times. Natural products with industrial applications can be produced from secondary metabolism of living organisms including plants, animals and microorganisms. The present study highlights the bioactivity of rhizosphere soil fungi Aspergillus ibericus from Ficus religiosa against opportunistic pathogens, through the production of secondary metabolite exhibiting antimicrobial activity. Antimicrobial metabolite obtained from A. ibericus was found to be effective against test microbes gram-positive bacteria, Bacillus subtilis, Staphylococcus aureus, Streptococcus mutans and Streptococcus pyogenes, gram-negative bacteria, Pseudomonas aeruginosa and Escherichia coli and yeasts such as Candida albicans and C. tropicalis. The antimicrobial metabolite was thus broad spectrum in nature. The effect of fermentation medium, temperature and pH was also observed on the production of antimicrobial metabolite.

7 illus, 6 tables, 32 ref

030578   KUSHVEER J S, MISHRA R, DEVADATHA B, KHAN M I K, SARMA V V (Biotechnology Dep, Pondicherry Univ, Pondicherry - 605 014, Email: veerkush1992@gmail.com) : Assessment of antioxidant, anti-lipid peroxidation, DNA damage protection and anticancer activities of marine mangrove fungi. Int J Pharm Biol Sci 2018, 8(4), 291-310.

Marine fungi are still a less explored group of microorganisms for novel medicinal properties that may have medicinal and pharmacological applications. In the present study, 55 marine fungi were screened for in-vitro antioxidant potential. Three marine fungi were shortlisted based on their higher antioxidant activity and were subjected to further studies including estimation of total phenolic & flavonoid content, qualitative screening of secondary metabolite groups and anticancer activity. Potential marine fungal isolates were identified and characterized based on their morphology and molecular phylogenetic analyses. The antioxidative properties of crude extracts derived from marine fungi were evaluated employing various in-vitro methods viz., DPPH, ABTS, nitric oxide radical scavenging assay, reducing power assay, anti-lipid peroxidation assay, oxidative DNA damage protection and cell viability assay. Among the three fungi, Marasmiellus sp. (PUK-64) showed highest antioxidant activity followed by Hypoxylon sp. (PUK-4) and Scedosporium aurantiacum (PUK-60). Marasmiellus sp. not only showed highest radical scavenging activity but also had highest phenolic and flavonoid contents. This study has demonstrated a notable antioxidant potential in the selected marine fungi in addition to appreciable anti-cancerous activity.

7 illus, 3 tables, 59 ref

030572   KANNAIYAN A (Biotechnology and Bioinformatics Dep, Bishop Heber Coll, Tiruchirappalli - 620 017, Email: aki.bioinfo84@gmail.com) : In silico docking studies of selective natural inhibitor against PB1 protein of influenza A (H1N1) virus. Int J Pharm Biol Sci 2018, 8(4), 214-9.

The influenza A (H1N1) virus, also termed as swine flu is a prominent source of disease and death since 2009. Currently there is a requirement to discover novel anti-viral drugs for incapacitating the increases. Conventionally, many herbal extracts of ginger, kalmegh, garlic, ajwain, menthe, tulsi, green tea and turmeric etc. has been used as progressive source of inhibition and cure of human influenza. The H1N1 polymerase protein composed of viral proteins PB1, PB2, and PA, accumulates with viral RNA and nucleoprotein (NP) to mediate transcription and replication of the viral genome. It is engaged amino acid sequence of H1N1 PB1 subjected to build the tertiary structure by modeler and validated by Ramachandran Plot, SAVES and ProSA server. Additionally, the build structure was subjected to dock with six natural compounds of herbs by Flexx software. Utmost of the selected compounds exposed more inhibitory activity against PB1 of H1N1 by binding. The current study furthermore shows the interaction of these selected compounds with residues of PB1 protein. Moreover, among six natural compounds, Curcumin found in turmeric detected to inhibit PB1(wild) protein of H1N1, this is intensely sustained by least binding energy (-13.0595) and Curcumin from turmeric, Caffeic acid and theaflavin from green tea was observed to inhibit PB1(mutated) protein of H1N1and it is strongly sustained by lowest binding energy (-17.8434, -15.6946 and -13.3391). In future, it might be give awareness to in vitro and in vivo evaluation study for Curcumin, Caffeic acid and theaflavin for further.

5 illus, 2 tables, 14 ref

030575   KHASA V, SINGH P, SIDHU P K (Biotechnology Dep, Deenbandhu Chhotu Ram Univ, Murthal - 131 039, Sonipat, Haryana, Email: vkhasasnp@gmail.com) : Plasmid profiling of Salmonella typhimurium isolates from commercial poultry farms of Haryana. Int J Pharm Biol Sci 2018, 8(4), 154-9.

Non-typhoidal salmonellosis is major cause of diarrhea in human and animals throughout the world leading to huge economic loss and mortality. Human Salmonellosis is sometimes traced back to animal source, majority of them from poultry and its products. Keeping this in mind, eleven strains of Salmonella typhimurium were isolated from different districts of Haryana; from broiler affected with Salmonellosis, which were revived and plasmid extraction was performed for profiling. The strains were classified into three plasmid types based on number and size of plasmids. Antibiotic sensitivity tests were also performed on all isolates and compared with plasmid. All plasmid types had 90kb plasmid whereas additional plasmid of size 110 kb, 120 kb and 23 kb were also observed in only two plasmid types. Plasmid type-1 and 2 had no major difference when checked for antibiotic resistance whereas plasmid type-3, having large size plasmids (110 and 120 Kbp) were found to exhibit resistance against the twelve antibiotics out of sixteen antibiotics used. Hence, high molecular weight plasmids may have role in antibiotic resistance and whereas plasmid between 50-100 kps may be serovar specific virulence genes.

1 illus, 1 table, 21 ref

030564   DWARAKANATH V, SHARATHCHANDRA R G (Biotechnology Dep, Tumkur Univ, Tumkur, Karnataka, Email: dwarak.vb@gmail.com) : Effect of Acacia farnesiana on SOD and CAT enzymes in ethanol induced gastric ulcer rats. Int J Pharm Biol Sci 2018, 8(4), 149-53.

Experimentation was conducted to test the hypothesis that Acacia farnesiana leaves has antioxidant activity. For this purpose, a study was designed using alcoholic extracts of Acacia farnesiana leaves produce significant antioxidant activity. 125 to 150 g weighing rats (Wistar albino) aging around one or two months of age are selected for this experimentation. The activity was tested in wistar albino rats at dose levels of 200 mg/kg, orally and compared with ranitidine (20 mg/kg) as standard. During the ulcer condition, there was a decline in the levels of Super oxide dismutase [SOD] and catalase [CAT]. This indicated that the generation of relative oxygen species during stress might be the causative factor for the inactivation of these enzymes. The methanol leaf extract of Acacia farnesiana has shown ulcer protective effect as dose dependently against ethanol induced gastric ulcer in rats. The said extract of Acacia farnesiana was found to decrease ulcer and acid pepsin secretion. A change was also seen in SOD, CAT, levels in rat gastric mucosa due to antioxidant property of alcoholic leaves extract of Acacia farnesiana. Antioxidant defense mechanism of the extract was probably due to metabolizing and scavenging H₂O₂.

3 illus, 2 tables, 10 ref

030586   PRAMILA R, RAMESH K V (Plant Biology and Plant Biotechnology Dep, Quaid-e- Millath Government Coll for Women, Chennai - 600 002, Email: ramesh.vijaya67@gmail.com) : Screening and identification of low-density Polyethylene (LDPE) degrading microorganism from municipal landfill soil. Int J Pharm Biol Sci 2018, 8(4), 53-5.

Low density polyethylene (LDPE) is a recalcitrant thermoplastic used in manufacturing of plastic bags, disposable cups and so on. It is non-degradable substance and it is the environmental pollution causing agent. The current research focused on evaluation of degradation of Low-density polyethylene (LDPE) by bacteria isolated from municipal dump soil by performing baiting technique clear zone plate assay and BATH test. We identified the bacterial strain as Bacillus species by gram staining and endospore staining method

3 illus, 2 tables, 12 ref

030593   SHIVAKUMAR R, VENKATARANGAIAH K, SHASTRI S, NAGARAJA R B, SHESHAGIRI A (PG Studies and Research in Biotechnology Dep, Kuvempu Univ, Shankaraghatta - 577 451, Email: krishnabiotech2003@gmail.com) : Antibacterial property and molecular docking studies of leaf calli phytochemicals of Bridelia scandens Wild. Pharmacogn J 2018, 10(6), 1221-9.

Bridelia scandens Wild. (Euphorbiaceae) leaves are widely used to cure asthma, bronchitis pleurisy, exudation, sores in mouth and genital cancers. To evaluate antibacterial activity of the leaf calli methanol extract (LCME). Mass production of leaf calli was established on MS medium supplemented with 0.5 mg/L BAP and 0.5 mg/L 2, 4-D. Methanol extract of the dried calli was subjected to HR-LCMS analysis, antibacterial screening of the extract was carried out against human pathogenic clinical isolates. Molecular docking study of HR-LCMS identified compounds was performed by docking with bacterial enzyme DNA gyrase. HR-LCMS analysis of LCME shows that the compounds azaperone bifonazole, fusidic acid, lasalocid and quinine as the major constituents. The antibacterial screening of LCME against clinical pathogens showed significant bactericidal activity against the strains Staphylococcus aureus (17.67 ± 0.88 mm.d.), Streptococcus pneumonia (13.67 ± 0.33), Pseudomonas aeruginosa (16.33 ± 0.67), Salmonella typhi (17.67 ± 0.33), and Vibrio cholera (15.33 ± 0.33) as compared to the standard drug ciprofloxacin. The molecular docking of lasalocid against the bacterial enzyme DNA gyrase exhibited good binding affinity of -4.9 kcal/mol, good drug likeness (2.5589), 2 hydrogen bonds and hydrophobic interaction with 7 amino acid residues, so that lasalocid processes good inhibitor as compared to other 4 compounds. LCME of Bridelia scandens showed significant antibacterial activity against Staphylococcus aureus and Salmonella typhi. Lasalocid is the major phytocomponent of LCME which exhibited good inhibitory activity against bacterial enzyme DNA gyrase. This investigation supported traditional claim of LCME as potential antibacterial drug.

3 illus, 5 tables, 33 ref

030591   SHARMA R K, GOEL A (Biotechnology Dep, GLA Univ, Mathura - 281 406, Email: sharmaanshul603@gmail.com) : Identification of phytoconstituents in Lawsonia inermis Linn. leaves extract by GC-MS and their antibacterial potential. Pharmacogn J 2018, 10(6), 1101-8.

Plant extracts contains multiple active constituents which leads to the production of new drugs from plants and chemicals derived from various parts of plants. The objective of present study was to investigate the GC-MS analysis and antibacterial activity of L. inermis leaves extracts. Crude methanol extract and its fractions were tested for the presence of active phytochemicals and GC-MS analysis of hexane; ethyl acetate and aqueous methanol fractions was performed. Antimicrobial activity against six bacterial strain’s Escherichia coli, Staphylococcus aureus, Bacillus subtelis, Salmonella typhi, Klebsiella and Pseudomonas aeruginosa was also tested. Phytochemical screening of extract confirmed the presence of carbohydrates, glycosides, quinones, steroids and phenol. In GC-MS chromatograms, 56, 108 and 19 peaks were obtained and out of these, 13, 17 and 7 compounds were identified in hexane, ethyl acetate and aqueous methanol fractions, respectively. For best of our knowledge in L. inermis leaves extract, Celidoniol and Vitamin E has not been reported earlier in hexane fractions. While 2, 3 dihydrobenzo furan, 1-H indole -1, 3 (2H)-dione, 1 (3H)-Isobenzofuranone, 1H Isoindole-1, 3 (2H) Dione, Napthelene, 2 ethoxy, 2 (4H) Benzofuranone, Vitamin E, Benzene, 1 isocyano 4 methyl are also identified for the first time in ethyl acetate fraction. Also, in aq. Methanol fraction 1(3H)-isobenzofuranone, Squalene and Vit E were not previously identified. Highest antibacterial activity was confirmed in crude methanol extract which might be due to all the antibacterial compounds present in its fractions. The present study helped in identifying phytoconstituents present in the extract and its fractions which are responsible for various biological and antibacterial activities.

3 illus, 6 tables, 66 ref

030594   SINGH A, VERMA N, KUMAR K (Biotechnology Dep, Multani Mal Modi Coll, Patiala - 147 001, Email: kuldeepbio@gmail.com) : L-asparaginase from Phyllanthus emblica (amla): A novel source. Int J Pharm Sci Res 2018, 9(12), 5394-400.

The biologically active compounds in plants reckoned their application in medicines from ancient times. Among the biologically active compounds enzymes have evolved as a potential therapeutic agent. Lasparaginase a therapeutic protein is extensively employed in medical sector to diagnose and treat leukemia. Bacteria, fungi, yeast and plants have been identified as potential sources of L-asparaginase. The side effects associated with microbial L-asparaginase restricted their utilization in treatment of leukemia and bought medicinal plants as source into focus. In the present study, different species of Phyllanthus genera have been screened for presence of L-asparaginase in different seasons. The leaves of Phyllanthus emblica with highest enzyme activity (20.3 IU/ml) and specific activity (5.2 IU/mg) emerged out as potential source for L-asparaginase production. Phyllanthus amarus and Phyllanthus niruri also contained appreciable amount of L-asparaginase. This is the first report of identifying presence of L-asparaginase in Phyllanthus genera. L-asparaginase from Phyllanthus emblica was further purified to homogeneity and the molecular mass of enzyme was found to approximately 85kDa. The yield of purified enzyme was 63.54 % with 8.5 fold increase in specific activity. Kinetic parameters, K_m and V_max of enzyme, were found to be 3.83 mM and 292.141 mM/min, respectively.

4 illus, 4 tables, 39 ref

030585   POORNIMA V, MAHENDRAN R, SUGANYA J, SHARANYA M (Bioinformatics Dep, VISTAS, Pallavaram - 600 117, Email: Poornivaasu.87@gmail.com) : Molecular docking studies of indenoisoquinoline derivatives with DNA topoisomerase I complex. Int J Pharm Sci Res 2018, 9(12), 5221-5.

The aim of the present study is to find the effectiveness of two series of indenoisoquinolines compounds against cancer. Topoisomerase I (Top 1) an essential enzyme that participates in the processes associated with separation of DNA strands and manages the super helical tensions through the transient breakage of one strand of duplex DNA, monitored by the unwinding of supercoiled DNA. The Top 1-DNA complex an effective target in the treatment of cancer was taken as a receptor for study. The two series of indenoisoquinolines compounds were taken as ligand molecules and the 3D structure of human Top1-DNA was retrieved from PDB database. The structures of ligand were drawn using Chemsketch. Docking studies were performed using GOLD which predicts the binding affinity and hydrogen bond interactions between the ligand and receptor. From the results, it was observed that the selected series of both nitrated indenoisoquinolines and 2, 3 dimethoxysubstituted indenoisoquinolines had better Gold score with the active site residue ARG364. Docking studies conclude both the series may act as potent inhibitor that can be further studied for developing anticancer agents.

2 illus, 1 table, 7 ref

030598   SUGANYA J, RADHA M, MANOHARAN S, POORNIMA V (Bioinformatics Dep, VISTAS, Chennai - 600 117, Email: suganyaj11@gmail.com) : In-silico analysis of SNPs from CAMP-GEFII gene associated with polycystic ovarian syndrome. Int J Pharm Sci Res 2018, 9(12), 5216-20.

Polycystic ovary syndrome (PCOS) is one of the most common disorders that occur in women at any age due to the endocrine hormone imbalance. The cause for this disorder is still not identified, but on recent research provided that disorder may be caused by some genetic variation. Predicting and understanding the effects of genetic variation that occurred in the gene are becoming more important for single nucleotide polymorphism to understand the molecular basis of genetic disease. From the literature survey, the candidate gene which is responsible for causing genetic PCOS was identified. In this work using computational methods, this candidate gene was analyzed completely to find out the genetic variation which in charge for altering the expression and the functional of the gene. On analyzing the gene, it was predicted that the protein which was translated from the gene played a key role for causing the major alteration in the gene. Using SNP analysis tool, further investigation were carried out to the disease causing protein and were predicted that the particular mutation occurred in the protein altered the function and structure of the gene. By using bioinformatics tool, an attempt was made to stop the mutation by replacing the original amino acid to the structure and sequence of the protein, which was suggested by the tools. Some clinical studies can be carried out further to confirm that the protein which was responsible for gene alteration in PCOS will function normally after some necessary modification are made in the protein which was suggested by computational methods.

1 illus, 5 tables, 26 ref

030587   RADHA M, SOUNDARYA R N, SUGANYA J, SARMA V R, MANOHARAN S, POORNIMA V, ANBARASU K (Bioinformatics Dep, Vels Institute of Science Technology and Advanced Studies, Chennai - 600 117, Email: mahenradha@gmail.com) : Molecular docking and molecular dynamics studies of quassinoids as HIV-1 TAT inhibitors. Int J Pharm Sci Res 2018, 9(12), 5210-5.

TAT (Trans-activator-Transcription Protein) is a viral protein encoded by the TAT gene in HIV-1-which is a lethal subtype of HIV (Human immunodeficiency Virus). It is vital for transcription of the viral genome. Previous studies show that in Human, TAT is a toxin-producing protein allowing cell death in obtained from QSAR studies, best quassinoid compounds were used to find the highest binding affinity compound by performing normal T-cells. Thereafter allows for progression towards AIDS (Acquired immunodeficiency syndrome). Traditionally, herbal medicines have played a vital role in the treatment of many diseases and ailments. Likewise, Quassinoids from the plant family Simaroubacaea, possess insecticidal, antimalarial and anticancer properties. Although studies have been conducted to find anti-HIV activities against other HIV-1 proteins, there are no traces of studies against Transactivator-Transcription protein (1JFW). The main objective of this study is to find an efficacious inhibitor against a synthetic HIV TAT protein (PDB- IJFW). After a thorough literature survey, the molecular and biological activity of quassinoid phytoconstituents has been recorded for QSAR (Quantitative structure-activity relationship) studies. Based on the results obtained from QSAR studies, best quassinoid compounds were used to find the highest binding affinity compound by performing docking analysis. Finally, the best compound with the highest binding affinity along with its measurement has been viewed in PYMOL. Furthermore, the aforementioned results from the docking studies were used to perform molecular dynamics simulation to confirm the efficiency of the compound against HIV-1.

4 illus, 3 tables, 17 ref

030600   SUJATHA A, SUBASH A (Biochemistry Dep, Avinashilingam Institute for Home Science & Higher Education for Women, Coimbatore - 641 043, Email: sujathaphdbt@gmail.com) : Isolation and purification of an extracellular protease from Bacillus subtilis ASASBT isolated from termite soil. Int J Pharm Sci Res 2018, 9(11), 4812-9.

Proteases are one of the most important enzymes in many industrial applications, particularly as additives in laundry detergent industry. Hence, the present study investigates the isolation and purification of an extracellular protease from the Bacillus subtilis ASASBT isolated from termite soil sample. Among the five protease producers, one of the isolates was selected for further studies. It was related to Bacillus subtilis on the basis of 16S rRNA gene sequencing and biochemical properties. The maximum protease activity recorded at 48 h of incubation at 37°C was 154.13 U/ml. Based on the maximum activity of the enzyme, further optimization studies were done with Bacillus subtilis ASASBT. Its optima pH and temperature of enzyme production were pH 7.0 and 40 °C, respectively. Starch and gelatin were the best carbon and nitrogen sources for the production of protease. The isolated protease was purified in four steps involving ammonium sulphate precipitation, dialysis, DEAE-Cellulose and sephadex G-100 chromatography had a 4.01 fold increase in specific activity (86.51 U/mg) and 23.73 % increase in recovery percentage. These properties suggest that protease from Bacillus subtilis ASASBT could find potential application in various industries.

9 illus, 3 tables, 30 ref

030595   SINGH S, GOPALAKRISHNAN S, KARTHIKEYAN N (Pharmaceutical Biotechnology Dep, Pharmacy Coll, SRIPMS - 641 044, Tamil Nadu, Email: keyanpharma@gmail.com) : Prevalence of AmpC β-lactamase in plasmid resistant gene from uti clinical isolates by molecular techniques. Int J Pharm Sci Res 2018, 9(11), 4784-93.

To detect the prevalence of Plasmid resistant AmpC β-lactamases gene in clinical isolates of gram negative organisms from UTI patients which produce resistance against multiple antibiotics. The gene coding for AmpC β-lactamases is also present in E. coli & Klebsiella species was not expressed because due to lack of promoter region, but the transfer of chromosomal genes to plasmids allows the expression of AmpC β-lactamases that hydrolyze the β-lactam ring, which has greater impact on resistance against multi-drug antibiotics, is a significant problem around the world. Among 20 non-repetitive clinical isolate of each Escherichia coli and Klebsiella pneumoniae, examined for identification and characterization of urine cultures based on morphological, biochemical tests, antibiotic resistant pattern, modifieddisc method and detection of AmpC gene by plasmid identification by agarose gel electrophoresis and amplification of AmpC gene by PCR techniques. The study detects the prevalence of AmpC gene primarily by modified disc inducer method as well as conformational molecular analysis by PCR amplification techniques. AmpC prevalence was observed in both strains from the UTI clinical isolates. The prevalence of AmpC resistance may differ due to the geographical variations was observed in different strains of gram negative organisms. The detection of AmpC resistance mechanism in plasmid DNA is an important factor to improve the clinical management of infection against higher antibiotics in relation with antibiotic resistance and cost of antibiotics which could help in therapeutics and UTI control process.

8 illus, 5 tables, 26 ref

030574   KAVITHA R (Biotechnology Dep, Periyar Univ PG Extension Centre, Dharmapuri - 636 701, Email: erokavi_vasu@yahoo.com) : Biochemical studies on the effect of ethanolic extracts of Trichosanthes dioica and Clitoria ternatea in streptozotocin induced diabetic male Wistar rats. Int J Pharm Sci Res 2018, 9(11), 4682-9.

To investigate the effect of ethanolic extracts of leaf and fruit of Trichosanthe dioica (T. dioica) and leaf of Clitoria ternatea (C. ternatea) on the biochemical parameters in streptozotocin (STZ) induced diabetic Wistar albino rats. Healthy male adult Albino rats of Wistar strain were randomly divided into eleven groups of six rats each were assigned into normal and diabetic groups (Group I to XI). Diabetes was induced in albino rats by administration of STZ (60 mg/kg i.p). Group I and II were kept as normal control and diabetic control. The other diabetic groups from group III to group X were treated with either individual and combined ethanolic extracts of T. dioica and C. ternatea at a dose of 200 and 400 mg/kg of body weight (bw) were administrated at a single dose per day and group XI was treated with glibenclamide at a dose of 600 μg/kg body weight for 28 consecutive days. The effect of ethanolic extracts of leaf and fruit of T. dioica and leaf of C. ternatea on blood glucose, serum insulin, glycosylated hemoglobin (HbA1c), urea, creatinine, SGOT, SGPT, LDH, ALP, total protein, albumin and globulin were evaluated in the diabetic rats. In the acute toxicity study, ethanolic extracts of leaf and fruit of T. dioica and leaf of C. ternatea were non-toxic and safe up to 2000 mg/kg body weight in rats. A significant decrease in blood glucose, HbA1c and other biochemical parameters levels and increase in serum insulin level were observed in diabetic rats treated with ethanolic extracts of T. dioica (leaf and fruit) and C. ternatea (leaf) compared to diabetic control rats. These findings suggest that the combined extract of T. dioica fruit and C. ternatea leaf have potent antidiabetic activity in STZ induced diabetic rats.

4 tables, 37 ref

030560   ABOELNOUR A, ME S Z, EL-KANNISHY S M H, KHEIR N A E (Clinical Pathology Dep, Mansoura Univ, Egypt, Email: aalaa_abo@yahoo.com) : Molecular study of intracellular adhesion genes (ica) and fibronectin binding protein genes (FnB) in clinical isolates of Staphylococcus aureus isolated from patients under chemotherapy. Int J Drug Dev Res 2018, 10(4), 8-12.

Staphylococcus aureus (S. aureus) is common hospital acquired pathogen. Biofilm formation capacity of S. aureus increases its pathogenicity. Biofilm formation is controlled by several factors among which are intracellular adhesion genes (ica) and fibronectin binding protein Genes (FnB). The aim of the present study is to evaluate the genotypic and phenotypic capacity of clinically isolated S. aureus strains for formation of biofilms. The study was conducted on 182 consecutive S. aureus isolates obtained from patients with hospital acquired infections. The isolates were subjected to antibiotics study by disc diffusion method, biofilm detection by microtitre plate method. Molecular studies for icaR, icaA, icaB, icaD and FnbA and FnbB was carried out by polymerase chain reaction. The frequency of biofilm formation among S. aureus was 35.5 %. The most frequent detected genes among S. aureus were icaR (68.2 %), icaC (63.6 %), ica D (60.6 %), fibA 56.1 %, fib (53.0 %, icaB, 51.5 % and icaA 30.3 %. Twenty one isolates of S. aureus had no detected genes of the studied seven genes. The present study reveals that biofilm formation was common our clinical isolates of S. aureus. The prevalence of icaABCDR genes and fib A, B were common among these isolates with predominance of icaR gene. Biofilm formation was associated with significant antibiotics resistance. Longitudinal studies are required to evaluate biofilm formation and its genetic mechanisms.

4 illus, 4 tables, 31 ref

030589   SARROPOULOU V, KRIGAS N, GRIGORIADOU K, MALOUPA E (Institute of Plant Breeding and Genetic Resources, Thessaloniki, Greece) : Asexual propagation and ex situ conservation of Hypericum empetrifolium Willd. subsp. empetrifolium (Hypericaceae), an east mediterranean medicinal plant with ornamental value. J Med Plants Stud 2018, 6(6), 235-41.

Hypericum empetrifolium subsp. empetrifolium is a medicinal plant of conservation concern and ornamental value restricted to Greece, Albania, Western Turkey and Cyrenaica in Libya. For vegetative propagation in mid-autumn, shoot tip softwood cuttings (5-6 cm) were immersed for 1 min in solutions of 4 IBA concentrations (0, 1000, 2000 and 4000 mg L^-1). Cuttings were placed in propagation trays in a peat: perlite (1:3) substrate, under mist. For in vitro propagation, shoot tips explants were cultured in MS medium. The effect of BA alone and combined with auxins on shoot proliferation was studied. For in vitro rooting, different auxins (IBA, NAA, IAA) applied at different concentrations were tested. Highest rooting percentage for cuttings was noticed when using 1000 mg L^-1 IBA (17.17 roots 2.84 cm long, 85.71 % rooting) (6 weeks). The 0.1 mg L^-1 BA + 0.01 mg L^-1 IBA hormonal combination was the best for in vitro shoot proliferation, yielding 5.5 shoots/explant, 100 % shoot formation and 5.42 shoot proliferation rate (4 weeks). Rooting in vitro was optimum with 0.1 mg L^-1 IBA (3.22 roots 1.05 cm long, 56.25 % rooting) (5 weeks). Acclimatization of rooted in vitro plants was successful exhibiting survival rate up to 90 %.

4 illus, 3 tables, 41 ref

030601   SUN X-Y, WANG Y-H, DONG Z-E, WU H-Y, CHEN P-P, XIE Q (Sun Yat-sen Univ, Guangzhou 510275, China, Email: xieq8@mail.sysu.edu.cn) : Identifying differential gene expression in wing polymorphism of adult males of the largest water strider: De novo transcriptome assembly for Gigantometra gigas (Hemiptera: Gerridae). J Insect Sci 2018, 18(6), 17.

Wing polymorphism is common in a wide variety of insect species. However, few studies have reported on adaptations in the wing polymorphism of insects at molecular level, in particular for males. Thus, the adaptive mechanisms need to be explored. The remarkable variability in wing morphs of insects is well represented in the water striders (Hemiptera: Gerridae). Within this family, Gigantometra gigas (China, 1925), the largest water strider known worldwide, displays macropterous and apterous males. In the present study, we used de novo transcriptome assembly to obtain gene expression information and compared body and leg-component lengths of adult males in different wing morphs. The analyses in both gene expression and phenotype levels were used for exploring the adaptive mechanism in wing polymorphism of G. gigas. After checking, a series of highly expressed structural genes were found in macropterous morphs, which were related to the maintenance of flight muscles and the enhancement of flight capacity, whereas in the apterous morphs, the imaginal morphogenesis protein-Late 2 (ImpL2), which might inhibit wing development and increase the body size of insects, was still highly expressed in the adult stage. Moreover, body and leg-component lengths were significantly larger in apterous than in macropterous morphs. The larger size of the apterous morphs and the differences in highly expressed genes between the two wing morphs consistently demonstrate the adaptive significance of wing polymorphism in G. gigas. These results shed light on the future loss-of-function research of wing polymorphism in G. gigas.

6 illus, 3 tables, 79 ref

030596   STEELE L D, COATES B S, SEONG K M, VALERO M C, MITTAPALLI O, SUN W, CLARK J, PITTENDRIGH B R (Entomology Dep, Illinois at Urbana-Champaign Univ, IL 61801, Email: steele11@illinois.edu) : Variation in mitochondria-derived transcript levels associated with DDT resistance in the 91-R strain of Drosophila melanogaster (Diptera: Drosophilidae). J Insect Sci 2018, 18(6), 1.

The organochloride insecticide dichlorodiphenyltrichloroethane (DDT) and its metabolites can increase cellular levels of reactive oxygen species (ROS), cause mitochondrial dysfunction, and induce apoptosis. The highly DDTresistant Drosophila melanogaster Meigen 1830 (Drosophila) strain, 91-R, and its susceptible control, 91-C, were used to investigate functional and structural changes among mitochondrial-derived pathways. Resequencing of mitochondrial genomes (mitogenomes) detected no structural differences between 91-R and 91-C, whereas RNAseq suggested the differential expression of 221 mitochondrial-associated genes. Reverse transcriptase-quantitative PCR validation of 33 candidates confirmed that transcripts for six genes (Cyp12d1-p, Cyp12a4, cyt-c-d, COX5BL, COX7AL, CG17140) were significantly upregulated and two genes (Dif, Rel) were significantly downregulated in 91-R. Among the upregulated genes, four genes are duplicated within the reference genome (cyt-c-d, CG17140, COX5BL, and COX7AL). The predicted functions of the differentially expressed genes, or known functions of closely related genes, suggest that 91-R utilizes existing ROS regulation pathways of the mitochondria to combat increased ROS levels from exposure to DDT. This study represents, to our knowledge, the initial investigation of mitochondrial genome sequence variants and functional adaptations in responses to intense DDT selection and provides insights into potential adaptations of ROS management associated with DDT selection in Drosophila.

2 illus, 2 tables, 133 ref

030605   YÖRÜK E, KELES E N, SEFER O, ERASLAN M (Molecular Biology and Genetics Dep, Istanbul Yeni Yuzyil Univ, Istanbul, 34010, Turkey, Email: emre.yoruk@yeniyuzyil.edu.tr) : Salinity and drought stress on barley and wheat cultivars planted in Turkey. J Environ Biol 2018, 39(6), 943-50.

The study was conducted to investigate the salinity and drought susceptibilities of barley and wheat varieties cultivated in Turkey by phenotypic assays and to evaluate the association of WRKY19 and WRKY52 transcription factors with salinity and drought stresses by gene expression assays. Salinity tests of treatment different concentration of (0, 0.5, 1 and 2 %) NaCl were managed with 13 barley and 22 wheat varieties germinated for 10 days. According to findings obtained from phenotypic investigations, two relative tolerant and two sensitive cultivars were selected and used in gene expression analysis. Transcript abundance for drought stress and relative fold changes for salinity stress were analyzed via expression assays of WRKY19 and WRKY52 genes. The minimum and maximum germination scores were changed between 0.090 ± 0.090 – 3.818 ± 0.400 (barley) and 0.454 ± 0.312 – 3.0913 ± 0.594 (wheat), while water loss rate (WLR) values ranged from 0.009 ± 0.0091 – 0.2 ± 0.0011 (barley) and 0.01 ± 0.0005 – 0.3 ± 0.1195 (wheat). In drought stress assessments, WRKY19 and WRKY52 transcripts abundances were relatively higher in relatively resistance cultivars in comparison to sensitive genotypes. Similarly, fold c h a n g e s i n g e n e expression were higher in resistant cultivars up to +26 changes. Drought and salinity stress factor analysis showed that there was no homogenous abiotic stress response profile for barley and wheat varieties in Turkey. According to gene expression analysis, WRKY19 and WRK52 genes could stimulate the drought and sal init y responses. This study is important in terms of analyzing the cereal varieties planted in Turkey, and providing an association between WRKY genes and abiotic stress.

2 illus, 2 tables, 30 ref

030581   MAHMUD S, NAZIR K H M N H, RAHMAN M T (Microbiology and Hygiene Dep, Bangladesh Agricultural Univ, Mymensingh 2202, Bangladesh., Email: tanvirahman@bau.edu.bd) : Prevalence and molecular detection of fluoroquinolone-resistant genes (qnrA and qnrS) in Escherichia coli isolated from healthy broiler chickens. Vet World 2018, 11(12), 1720-4.

The present study was carried out to determine the prevalence and molecular detection of fluoroquinolone-resistant Escherichia coli carrying qnrA and qnrS genes in healthy broiler chickens in Mymensingh, Bangladesh, and also to identify the genes responsible for such resistance. A total of 65 cloacal swabs were collected from apparently healthy chickens of 0-14 days (n = 23) and 15-35 days (n = 42) old. The samples were cultured onto Eosin Methylene Blue Agar, and the isolation and identification of the E. coli were performed based on morphology, cultural, staining, and biochemical properties followed by polymerase chain reaction (PCR) targeting E. coli 16S rRNA genes. The isolates were subjected to antimicrobial susceptibility test against five commonly used antibiotics under fluoroquinolone (quinolone) group, namely gatifloxacin, levofloxacin, moxifloxacin, ofloxacin, and pefloxacin by disk diffusion method. Detection of qnrA and qnrS genes was performed by PCR. Among the 65 cloacal samples, 54 (83.08 %) were found to be positive for E. coli. Antibiotic sensitivity test revealed that, of these 54 isolates, 18 (33.33 %) were found to be resistant to at least one fluoroquinolone antibiotic. The highest resistance was observed against pefloxacin (61.11 %). By PCR, of 18 E. coli resistant to fluoroquinolone, 13 (72.22 %) were found to be positive for the presence of qnrS. None of the isolates were found positive for qnrA. Fluoroquinolone-resistant E. coli harboring qnrS genes is highly prevalent in apparently healthy broiler chickens and possesses a potential threat to human health.

2 illus, 5 tables, 24 ref

030579   LOUNAS A, OUMOUNA-BENACHOUR K, MEDKOUR H, OUMOUNA M (Medicine and Animal Surgery Dep, University of Blida, Algeria, Email: azizovet@yahoo.fr) : The first evidence of a new genotype of nephropathogenic infectious bronchitis virus circulating in vaccinated and unvaccinated broiler flocks in Algeria. Vet World 2018, 11(11), 1630-6.

Avian infectious bronchitis virus (IBV) frequently infects broilers and is responsible for severe economic losses to the poultry industry worldwide. It has also been associated with kidney damage in the broiler flocks. The aim of the present study is to determine the presence of IBV and its possible involvement in kidney damage of broiler chicks. 14 clinically diseased broiler flocks from Western and Central Algeria were sampled and analyzed by hemagglutination inhibition (HI) test and reverse transcriptase-polymerase chain reaction (RT-PCR) followed by phylogenic analysis. The QX (100 %) and 4/91 (60 %) IBV serotypes were the most prevalent in the kidney damaged broilers regardless of vaccination status. The molecular detection of avian IBV by RT-PCR identified six samples as positive, of which only two isolates were typable by sequencing. We identified a novel IBDZ13a genotype which showed 93 % sequence homology to the partial-S1 gene sequence of the IB 4/91 commercial vaccine strain. Sequencing analysis characterized this virus as a novel and divergent IB 4/91 field virus with eight amino acid substitutions that might have resulted in altered immunogenicity. The isolation of a new IBV strain (IBDZ13a) from vaccinated broiler flocks may explain the failure of the vaccination programs against IBV field strains. Combination of the HI test and RT-PCR indicated that the nephropathogenic IB outbreaks in broilers are related to this novel strain.

2 illus, 3 tables, 36 ref

030562   DHAR P K, DUTTA A, DAS A, JALAL M S, BARUA H, BISWAS P K (Microbiology and Veterinary Public Health Dep, Chittagong Veterinary and Animal Sciences Univ, Chittagong-4225, Bangladesh, Email: pangkaj23@gmail.com) : Validation of real-time reverse transcription polymerase chain reaction to detect virus titer and thermostability of Newcastle disease live virus vaccine. Vet World 2018, 11(11), 1597-603.

Newcastle disease is one of the most common diseases affecting poultry in Bangladesh. The disease can cause up to 100 % mortality but is preventable if birds are timely and properly vaccinated with a vaccine of standard virus titer. Different live vaccines are commercially available in the country - most, if not all, are produced using lentogenic strains of the virus with variable virulence. One of the disadvantages of these vaccines is that they are not stable at high environmental temperature, and therefore, a proper cold chain must be maintained during transportation and storage. Information on how long these vaccine viruses can withstand environmental temperature, which is near the vicinity of 37 $°$C in the summer season in Bangladesh, is scanty. The aim of this research was to measure the effect of temperature on virus titer of live ND virus vaccines and to develop a real-time reverse transcription polymerase chain reaction (rRT-PCR) standard curve to indirectly determine hemagglutination (HA) titer of virus by this highly sensitive method. In this study, thermostability of five commercial live vaccines containing LaSota, F, Clone 30, and B1 type LaSota strains was observed for up to 35 days keeping them at 37 ºC. From the most thermostability yielding sample, two rRT-PCR standard curves were developed: (1) By plotting the cycle threshold (C_T) values as obtained from 10-fold serial dilutions up to 10⁻³ against their corresponding log (to the base 10) dilutions and (2) by plotting the C_T values obtained from serial HA dilutions up to 2⁻⁴ against their corresponding HA titer dilutions. The PCR efficiencies based on which the graphs were fitted were also evaluated. The vaccine from the LaSota strain withstood 37 ºC for 35 days with a gradual declination of HA titer over time, and this vaccine also had the highest initial HA titer, which was 2¹¹. The vaccine made from F strain was inactivated quickly, and it had the lowest HA titer at the beginning of the study. The first standard curve developed can be used to assess the level of virus titer in a diluted sample compared with the titer in the original undiluted vaccine preparation by plotting the C_T value obtained from the dilution by rRT-PCR. The second standard curve can be used to calculate the HA titer of a vaccine dilution by plotting the C_T value as obtained from the dilution by rRT-PCR. The regression equations for the first and second graphs were y = −3.535x + 14.365 and y = −1.081x + 13.703, respectively, suggesting that, for every 3.53 cycles, the PCR product would have increased 10 times and 2 times for every 1.08 cycles, respectively, indicating nearly (but not exactly) 100 % PCR efficiency.

4 illus, 2 tables, 31 ref

030563   DINANI O P, TYAGI P K, MANDAL A B, TYAGI P K, POPAT D S, TIWARI S P (Avian Nutrition and Feed Technology Div, ICAR-Central Avian Research Institute, Uttar Pradesh, Email: dr_dinani@rediffmail.com) : Effect of feeding rice based distillers dried grains with solubles and gluten meal on nutrient transporter genes and immunity in broiler chickens. Vet World 2018, 11(11), 1592-6.

The aim of this study was to investigate the effect of feeding rice based distillers dried grains with solubles (rDDGS) and gluten meal on nutrient transporter genes and immunity in broiler chickens. A 2×3 factorial design resulted in six experimental diets, namely T1 (no rDDGS/rice gluten meal [RGM]/enzyme), T2 (no rDDGS/RGM, with multienzymes), T3 (12.5 % rDDGS, 15 % RGM, no enzyme), T4 (12.5 % rDDGS, 15 % RGM, with protease enzyme), T5 (10 % rDDGS, 12.5 % RGM, no enzyme), and T6 (10 % rDDGS, 12.5 % RGM, with protease enzyme). Each treatment was allocated five replicates of chicks, with eight birds in each. Nutrient transporter genes such as Mucin (MUC 2), excitatory amino acid transporter 3 (EAAT3), and peptide transporter (PepT1) and immunity were estimated using standard procedures. Feeding rDDGS and RGM combination improved humoral immunity, while cell-mediated immunity did not show any significant (p > 0.05) effect on broiler chickens. MUC and PepT1 genes showed significantly (p < 0.01) decreased relative fold expression in 12.5 % rDDGS +15 % RGM combination, while EAAT3 gene showed significantly (p < 0.01) decreased relative fold expression in both rDDGS and RGM combination levels. Thus, it may be concluded that feeding rDDGS and RGM combination improved humoral immunity but had an adverse effect on nutrient transporter gene in broiler chickens.

3 tables, 12 ref