Suan Sunandha Science and Technology Journal Editorial Board Editor in Chief Asst. Prof. Dr. Anat Thapinta Associate Editors Asst. Prof. Dr. Sirilak Namwong Dr. Tatsanawalai Utarasakul Asst. Prof. Dr. Yuttana Sudjaroen Dr. Sansanee Sansiribhan Dr. Narumon Boonman Mr. Dulyawit Prangchumpol Dr. Pantip Kayee Mrs. Kittiya Poonsilp Dr. Pratheep Meewattana Mr. Michael Cota Assistant Editors Dr. Serisa Pinkham Miss Nunyong Khamparat Dr. Rutanachai Thaiprathum Miss Patthama Hirunyophat Miss Kunya Bowornchokchai Miss Tidarat Sanphom Mr. Noppadon Chamchoi Software and Technical Support Miss Charinthip Hirannukhrao Mrs. Lalisa Sahanawin Mr. Dulyawit Prangchumpol Mr. Michael Cota Mrs. Kittiya Poonsilp Editorial Liaisons Miss Charinthip Hirannukhrao Mrs. Lalisa Sahanawin The Suan Sunandha Science and Technology Journal (ISSN 2351-0889) is officially published semi-annually by the Faculty of Science and Technology, Suan Sunandha Rajabhat University, 1 U-thong Nok Road, Dusit District, Bangkok 10300. Printed by Podduang Enterprise Co., Ltd., Bangkok Thailand Editorial Board Asst. Prof. Dr. Kheamrutai Thamaphat Assoc. Prof. Dr. Thi Thanh Van Ngo Department of Physics, Faculty of Science, Hanoi Water Resources University, Vietnam King Mongkut's University of Technology Thonburi Assoc. Prof. Dr. Chartchai Leenawong Prof. Dr. L. Lee Grismer Department of Mathematics, Faculty of Science, King Departmet of Biology, La Sierra University, USA Mongkut's Institute of Technology Ladkrabang Dr. Noppadon Kitana Assoc. Prof. Dr. Peraphon Sophatsathit Department of Biology, Faculty of Science, Faculty of Science, Chulalongkorn University Chulalongkorn University Assoc. Prof. Dr. Worasit Choochaiwattana Dr. Sittiruk Roytrakul Faculty of Information Technology, National Center for Genetic Engineering and Dhurakij Pundit University Biotechnology (BIOTEC) Prof. Dr. Somboon Tanasupawat Asst. Prof. Dr. Tosak Seelanan Faculty of Pharmaceutical Sciences, Department of Botany, Faculty of Science, Chulalongkorn University Chulalongkorn University Prof. Dr. Vithaya Meevootisom Asst. Prof. Dr. Boon-ek Yingyongnarongkul Department of Microbiology, Faculty of Science, Department of Chemistry, Faculty of Science, Mahidol University Ramkhamhaeng University Assoc. Prof. Nongluksna Sriubolmas Dr. Prasat Kittakoop School of Pharmacy, Eastern Asia University Chulabhorn Research Institute Asst. Prof. Dr. Suthep Wiyakrutta Dr. Naruebodee Srisang Department of Microbiology, Faculty of Science, King Mongkut's University of Technology Ladkrabang Mahidol University Prof. Dr. Ruide Yu Dr. Warapong Tungittiplakorn Xinjiang Institute of Ecology and Geography, Department of Agro-Industrial Technology, Chinese Academy of Sciences, China Faculty of Applied Science, King Mongkut's University of Technology North Prof. Dr. Subhash C. Pandey Bangkok Editor-In-Chief, Journal of Environmental Research and Development, Bhopal Assoc. Prof. Dr. Noppawan Piaseu Faculty of Medicine, Ramathibodi Hospital, Assoc. Prof. Dr. Benjaphorn Prapagdee Mahidol University Faculty of Environment and Resource Studies, Mahidol University Asst. Prof. Dr. Wararit Panichkitkosolkul Department of Mathematics and Statistics, Faculty Asst. Prof. Dr. Srilert Chotpantarat Science and Technology, Thammasat University, Geology Department, Faculty of Science Rangsit Center Chulalongkorn University SUAN SUNANDHA SCIENCE AND TECHNOLOGY JOURNAL Suan Sunandha Rajabhat University, Faculty of Science and Technology 1 U-thong Nok Road, Dusit District, Bangkok 10300 THAILAND CONTENTS Volume 1, Number 1 July 2014 Detection of Aflatoxin Producing Aspergillus flavus in Post-harvest Contaminated Vigna ungulculata Seeds Ajay Kumar Gautam___________________________________________________________1-4 Sustainability: an Approach in Planning to Raise the Quality of Life Through Open Space Development Sonal Y. Khobragade__________________________________________________________5-13 Interactive e-Learning System for Thalassemia: a Case of a University Hospital in Thailand Phanu Waraporn____________________________________________________________ 15-19 Wild Elephant Conservation Using Sound Waves to Obstruct Them from Plantations: a Case Study at Kui Buri District, Thailand Pratheep Meewattana and Chollathorn Chammarnkid_______________________________21-24 Bird Diversity in Fruit Gardens in Bang Nang Li Sub-district, Amphawa District, Samut Songkram Province Nitinarth Charoenpokaraj_____________________________________________________25-28 Suan Sunandha Journal of Science and Technology Vol. 1 No. 1 © 2014 Faculty of Science and Technology, Suan Sunandha University Detection of Aflatoxin Producing Aspergillus flavus in Post-harvest Contaminated Vigna ungulculata Seeds Ajay Kumar Gautam Department of Botany, Abhilashi Institute of Life Sciences, Mandi-175008 (H.P.) India. Abstract: The present study was carried out with a specific objective to study post- harvest spoilage of Lobhiya (Vigna unguiculata) seeds contaminated with Aspergillus flavus. Infected seeds were collected and cultured on potato dextrose agar (PDA) media, at 25±2 °C. Aspergillus flavus isolates were primarily characterized by its morphological and microscopic characteristics. Collected fungal isolates were also screened for their afaltoxigenic nature on preliminary basis and at molecular level. For preliminary screening, 5 mm disc of fungal culture was soaked with few drops of liquid ammonia. Color change from yellow pigment to plum-red with different intensities showed the mycotoxic nature of the fungus. DNA from fungal isolates was isolated and amplified using PCR with aflatoxin specific primers, apa-2, ver-1 and omt-1. Amplicons of 1032 bp, 895 bp and 596 bp were obtained in most of the isolates regardless of primer set used which was useful to differentiate between mycotoxic and nontoxic isolates of A. flavus. The isolation of aflatoxigenic strains of A. flavus during post-harvest period of lobhiya seeds raise a serious concern over the quality of seeds and a threat to heath of consumers. It was concluded that Aspergillus flavus is responsible for postharvest spolilage of Lobhiya (Vigna unguiculata). Keywords: Vigna unguiculata (L.) Walp.), post-harvest spoilage, Aspergillus flavus, aflatoxins. Introduction Materials and Methods Lobhiya (Vigna unguiculata (L.) Walp.) is an edible legume plant and an important bean of The Lobhiya seeds showing typical symptoms family Fabaceae, cultivated globally for its of fungal association were collected from nutritive value and edible importance. The seeds different location of the Himachal Pradesh, a are pale-colored with prominent black spot, so North Indian state. About 30 samples were known as black eyed pea or bean. Nutritionally, collected from all the locations. To clarify this the lobhiya seeds are energy rich, having a very fungus, collected seed samples were carried to good concentration of carbohydrates, fats, laboratory and examined carefully. Infected seeds proteins, vitamins and essential micro and macro were observed under hand lenses and dissecting elements (Davis et al., 1991). microscope for morphological characteristics of In September - October 2013, local associated pathogen. With the help of sterilized agricultural fields of Himachal Pradesh were needle or spatula, yellowish green fungal surveyed for the post-harvest spoilage of Lobhiya pathogen was cultured aseptically on Potato seeds. A clear-cut yellowish green fungal Dextrose Agar (PDA) media, and incubated at infection was noticed on the seed surface. 25±2°C in darkness for 4-5 days. The fungus Therefore, the present study was conducted to grown on culture media was isolated consistently ascertain the strains of fungi isolated from post and characterized primarily by its morphological harvested seeds of V. unguiculata. Collected and microscopic characteristics. fungal isolates were also screened for their The mycotoxin producing potential of mycotoxic nature. collected isolates of Aspergillus flavus is *Corresponding Author: [email protected] 1 Suan Sunandha Journal of Science and Technology Vol. 1 No. 1 _______________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________________ preliminarily carried out as per the method symptoms on Lobhiya seeds. Initially, the fungus described by Saito and Machida (1999). A 5mm appeared as yellowish-green on seeds surface that disc of fungal culture was soaked with few drops invaded healthy seeds later on (Fig.1 A-B). On of liquid ammonia and observe disc for the color PDA medium, colonies were yellowish-green, changed. entire margin with umbonate elevations, exudate The aflatoxin producing nature A. flavus absent, reverse colorless to yellow and moderate isolates was also studied at molecular level. DNA to rapid in growth. Mycelium white, branched, from fungal isolates was isolated following septate; conidiphore 600 to 800 µm in length, 15 CTAB method (Shapira et al., 1996; Konietzny to 20 µm in diameter, vesicle globose to and Geriner, 2003) and amplified using PCR with subglobose; conidia 20 to 45 µm in size, aflatoxin specific primers, apa-2, ver-1 and omt-1 yellowish green, smooth; phialides covering (Shapira et al., 1996; Konietzny and Geriner, nearly entire vesicle, biseriate primary and 2003). secondary phialides 7 to 10 µm in size, cleistothecia observed. On the basis of Results