In vitro Culture and Diet of Entomopathogenic Nematodes (original) (raw)
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Mass production of native isolate of entomopathogenic nematode, Steinernema sp. (AAU St-1
current nematology, 2021
The in vivo mass production of native EPN, Steinernema sp. (AAU St-1) was carried on different instars of the rice moth (Corcyra cephalonica). The maximum average yield of 90307 IJs/larva of Steinernema sp. (AAU St-1) was recorded from the fifth instar, followed by 78674 and 47771 IJs per larva from the fourth, third instar, respectively.
A total of 120 soil samples were collected from different habitats, 9 samples (7.5%) were found to be positive for EPN. The highest frequency of occurrence of EPNs (58.3%) was recorded in Ber orchard with 7 samples infected for EPNs followed by citrus and cactus (8.3%). Amongst them, the frequency of occurrence of Steinernematid and Heterorhabditid nematodes were 41.7 and 16.7 per cent, respectively. G. mellonella larvae infected with EPN of the genus Heterorhabditidae may have reddish-orange coloring while those infected with Steinernema spp. show gray-brown coloring. As the inoculum level increased, there was an increase in progeny production of both S. abaasi and H. indica.
Mealworm Powder as Culture Media of Local Isolate Semarang Entomopathogenic Nematodes
Biosaintifika: Journal of Biology & Biology Education, 2018
Many researchers confirmed that entomopathogenic nematodes (EPNs) of the genera Heterorhabditis and Steinernema can be cultivated in vitro using artificial media that containing animal nutrition. However, artificial media with insect component hasn't been widely studied. This research aims to analyze the population of EPNs isolate of Semarang cultivated in mealworm powder media. Five doses of mealworm powder (Tenebrio molitor) were tested in this research, i.e: 0.5; 1.0; 1.5; 2.0 g. Culture media using 1 g of mealworm larvae was used as control. The best treatment was further tested for its pathogenicity on Macrotermes sp. at seven levels of invective juveniles (IJs) : 0; 50; 100; 150; 200; 250; 300 IJs/mL. Each treatment was repeated five times. The EPNs population and termites mortality were analyzed using ANAVA, whereas pathogenic value was calculated using Probit analysis. The result showed EPNs population were significantly (LSD test; α> 0.05), likewise on termites mortality. The EPNs isolate of Semarang optimally at 0.5 g mealworm powder and pathogenicity against termites based on LD 50 and LD 90 values at 220 JI/mL and 410 JI/mL doses, respectively. In conclusion, this result can be an alternative to mass cultivation of EPNs, in effort of development of local bioinsecticides. The findings of this study also inform farmers that EPNs can be easily cultivated using the simple and available materials.
Effect of the inoculum size on the axenic in vitro production of Steinernema carpocapsae
Ciencia e investigación agraria, 2012
Effect of the inoculum size on the axenic in vitro production of Steinernema carpocapsae. Cien. Inv. Agr. 39(1): 137-145. The effect of inoculum size on the axenic in vitro production of Steinernema carpocapsae, using a transparent medium with hemoglobin and cholesterol, was studied. Axenic cultivation was performed with fertile eggs of S. carpocapsae (hatching of eggs: 89 ± 4%), which were obtained by lyses of female nematodes and subsequent incubation. Two important aspects of this work include the use of a completely transparent medium enriched with hemoglobin and cholesterol and the high multiplication factor of the population (MFIP) of 87, which is considered high for in vitro axenic cultures of nematodes. In vivo methods have the advantage of high MFIP, but they also have the disadvantage of the high number and complexity of the operations involved for the purposes of harvesting axenic nematodes. It is assumed that the availability of a high number of nematodes in a transparent medium will further encourage the characterization of new native strains of nematodes.
Mass Production of Entomopathogenic Nematodes or Plant Protection-A Review
Chemical pesticides may cause secondary pest outbreaks, accelerate the development of resistance, destroy natural enemies and create hazards for human's poisonings and approximately twenty thousand deaths occurring annually due to pesticides. As a result, regulatory agencies in different countries have imposed restrictions on chemical pesticide usage. Biological control using predators, parasitoids or pathogens can be an effective measure in insect pest management. Research and development on biological control and insect pests has increased manifold during the past two decades and entomopathogenic nematodes (EPN's) have been developed as one of the tool in insect management. Already, the EPN based biopesticides occupy the major share in the biopesticide world, being second only to Bacillus thuringiensis. EPN's of the families Steinernematidae and Heterorhabditidae (Order: Rhabditida) are lethal parasites associated with symbiotic bacteria of the family Enterobacteriaceae. Steinernematids are associated with Xenorhabdus spp. and Heterorhabditids with Photorhabdus spp. EPN's can be mass produced by in vivo or in vitro methods. The use of EPN is safe for both the user and the environment.
Mass Production of Entomopathogenic Nematodes- A Review
International Journal of Environment, Agriculture and Biotechnology, 2018
Utilization of entomopathogenic nematodes (EPNs) is an ecofriendly method of crop protection. EPNs can be easily mass produced. Production approaches are either in vivo or in vitro methods (solid and liquid). Most nematodes intended for commercial application are produced in solid or liquid fermentation technology. However, for laboratory research and small greenhouse or field trials, in vivo production of entomopathogenic nematodes is the common method of propagation. Mass production of EPNs is influenced by the amount of progeny required, time, resources, the costs of production, as well as the level of expertise available. The differences in nematode life cycle and bacterial symbiosis play major role in final nematode yields. This review describes the general biology of EPNs and gives an overview of studies to date on EPNs mass production. Keywords-Entomopathogenic nematodes, bacterial symbiosis, biocontrol agent, in vivo mass production, in vitro mass production. I.
Nematological Research (Japanese Journal of Nematology), 2016
To establish an easy axenic culture method for Steinernema carpocapsae and to understand the nutrition provided for nematode growth and propagation produced in the insect cadaver, we tested several supplementations to axenic liquid cultures. S. carpocapsae grew but reproduced poorly on the liquid static axenic culture developed for bacterial-feeding nematodes; however, supplementation by S. carpocapsae-infected insect cadavers sterilized by autoclaving largely improved nematode growth and propagation. These results suggest supplementation of the cadaver is a useful method to improve axenic culture and the autoclaved nematodeinfected insect contains important heat-stable nutritional factors for the g row th and reproduction of S. carpocapsae. Nematol. Res. ()-().
The present study deals with the batch and fed-batch mass production of Steinernema carpocapsae. S. carpocapsae is an entomoparasitic nematode that is used as a biological control agent of soil-borne crop insect pests. The ability and efficiency of fed-batch culture process was successful through the utilization of the nematode's bacterial symbiont Xenorhabdus nematophila. Results from the fed-batch process were compared to those obtain from the standard batch process. The fed-batch process successively improved the mass production process of S. carpocapsae employing liquid medium technology. Within the first week of the fed-batch process (day six), the nematode density obtained was 202,000 nematodes mL −1 ; whereas on day six, batch culture mode resulted in a nematode density of 23,000 nematodes mL −1. The fed-batch process was superior to that of batch production with a yield approximately 8.8-fold higher. In fed-batch process, the nematode yield was improved 88.6 % higher wit...