~ 1207 ~ The Pharma Innovation Journal 2022; SP-11(12): 1207-1210 ISSN (E): 2277-7695 ISSN (P): 2349-8242 NAAS Rating: 5.23 TPI 2022; SP-11(12): 1207-1210 © 2022 TPI www.thepharmajournal.com Received: 19-09-2022 Accepted: 25-10-2022 SV Dobariya M.Sc. (Agri.) Student, Department of Nematology, BACA, AAU, Anand, Gujarat, India Ajay Kumar Maru Assistant Professor, Department of Nematology, BACA, AAU, Anand, Gujarat, India RK Thumar Professor and Head, Department of Nematology, BACA, AAU, Anand, Gujarat, India Corresponding Author: SV Dobariya M.Sc. (Agri.) Student, Department of Nematology, BACA, AAU, Anand, Gujarat, India Isolation and mass production of native isolates of entomopathogenic nematodes from Anand (Gujarat), India SV Dobariya, Ajay Kumar Maru and RK Thumar Abstract The survey was conducted in Anand city and its adjoining area during Kharif-2021 to isolate native entomopathogenic nematodes (EPNs). A total of 94 soil samples were collected from fruit plants, perennial trees and fallow land, out of them two samples were found positive for EPNs, one from the campus of the Bhikhabhai Jivabhai Vanijya Mahavidhyalaya (Palm) and the second one from Atmiya Vidhya Dham (fallow land). The frequency of occurrence of these nematodes was 2.13 per cent. Based on primary identification up to the genera level, the first isolate was identified as Heterorhabditis sp. (Anand H-I). and the second was Steinernema sp. (Anand St-I). Both isolates were mass multiplied on six different artificial media. The maximum multiplication of Heterorhabditis sp. (Anand H-I) and Steinernema sp. (Anand St-I) was observed on Wout’s medium followed by Soya flour medium for Heterorhabditis sp. (Anand H-I) and Bengal gram medium for Steinernema sp. (Anand St-I). Keywords: Entomopathogenic nematodes, EPNs, survey, isolation, mass production, Heterorhabditis sp. and Steinernema sp. Introduction Entomopathogenic nematodes (EPNs) are parasites of insects belonging to the families Steinernamatidae and Heterorhabditidae. The infective juveniles of these families are free- living, non-feeding and have the ability to search out their hosts. This group of nematodes is characterized by carrying specific symbiotic bacteria of the genus Xenorhabdus or Photorhabdus in their intestine which play an important role in the pathogenicity of the nematode-bacteria complex to insect hosts and the subsequent reproduction of the nematodes in the hosts. They have the potential for long-term establishment in the soil through the recycling of infected insect larvae. The EPNs possess unique attributes like complex nematode-bacterium mutualistic symbiosis, broad host range and their amenability to be cultured in in-vivo and in-vitro on large scale, making them very suitable to be used as a biocontrol agent. EPNs are currently used as bio-pesticides for controlling several important insect pests worldwide (Shapiro-Ilan et al., 2002) [13] . EPNs-based pesticides are nowadays largely marketed in western countries in comparison to other organisms, apart from the bacteria Bacillus thuringiensis, but the survival and adaptability of EPNs are influenced by environmental factors. Due to unfavorable climatic conditions, exotic EPN strains may not be well adapted to local climates and their efficacy might be reduced. Therefore, isolation of native EPNs strain and mass multiplication is very important. This study aims to isolate and in vitro mass multiply the native strain of EPNs isolated from Anand city. Materials and Methods Rearing of bait insect, Corcyra cephalonica: In the present study, Rice moth Corcyra cephalonica larvae was used for the mass culturing of EPN isolates. Rice moth C. cephalonica larvae were obtained from AICRP on Biological Control of Crop Pests, ICAR unit-9, AAU, Anand. The larvae were reared on artificial diet containing following composition sorghum whole grain – 2.5 kg, broken sorghum grain – 0.5 kg, Streptomycin – 0.5 g and yeast powder – 1.0 g. Collection and baiting of soil samples: A total of 94 soil samples were collected from different localities of Anand city and its adjoining area from fruit plants, perennial trees and fallow land from the depth of 10-15 cm during Kharif 2021 as per the methodology described by Kaya and Stock (1997) [7] .