Biological Control Technology Utilizing Heterorhabditis bacteriophora and Steinernema carpocapsae (original) (raw)

Mass Production of the Beneficial Nematode Steinernema carpocapsae Using Solid State Fermentation

Journal of Advanced Agricultural Technologies, 2018

Steinernema carpocapsae is a microscopic entomopathogenic nematode (EPN) that may be used as an alternative to chemical pesticide. This species creates a symbiotic relationship with the bacteria Xenorhabdus nematophila. This biological control agent has many advantages compared to chemical pesticides as it does not harm either the environment or humans. Steinernema carpocapsae is a vector for the bacteria to infect the targeted insect pest. The bacteria kills the host within 24-48 hours. This paper focuses on the mass production of beneficial nematodes using solid state fermentation. The purpose of the experiment was to find the optimum conditions to mass produce the nematode efficiently. Maximizing yield with the minimalized nutrients will increase the cost efficiency of production, making it a more affordable attractive alternative to harmful chemical pesticides. 

A Low-Cost Technology for Entomopathogenic Nematode Large-Scale Production

Multiphase Bioreactor Design, 2001

Entomopathogenic nematodes of the genera Steinemema and Heterorhabditis may provide a valuable alternative to chemical insecticides. The characteristics that make them excellent biopesticides include their wide host spectrum, the ability to search for and kill hosts rapidly, and their high virulence and reproductive rates. Furthermore, they are considered environmentally safe. The major constraint to overcome before the onset of commercialisation is their mass production. Entomopathogenic nematodes are currently mass-produced in vivo or in vitro, either in solid culture or in liquid cultivation. An overview of these mass production methods and an analysis of three different bioreactor designs are presented. The progress achieved in liquid culture due to an improvement on sexual contact between adults (better mixture of the solid phase), which results in higher yields (RF), as compared with those reported before, is demonstrated. This improvement in the area of bioreaction engineering allowed these biopesticides to become more competitive compared to chemical insecticides. However, further technological advances and biological studies towards a better understanding of physiology and genetics of the complex nematode-bacterium are still required.

Asean Economic Biopesticide : Production of Biopesticide Entomopathogenic Nematodes for Biological Control Insect Pests for Organic Farming

2014

Entomopathogenic nematodes of the genera Heterorhabditis and Steinernema are commercially used to control pest insects. They are symbiotically associated with bacteria of the genera Photorhabdus and Xenorhabdus, respectively, which are the major source for the nematodes. The biology of the nematode-bacterium complex is described, a historical review of the development of in vitro cultivation techniques is given and the current use in agriculture is summarised. Media development is mainly directed towards cost reduction, as the bacteria are able to metabolise a variety of protein sources to provide optimal conditions for nematode reproduction. The process technology is described, discussing the influence of bioreactor design and process parameters required to obtain high nematode yields. As two organisms are grown in one vessel and one of them is a multicellular organism, the population dynamics and symbiotic interactions need to be understood in order to improve process management. ...

Mass production of entomopathogenic nematodes for plant protection

Applied Microbiology and Biotechnology, 2001

Entomopathogenic nematodes of the genera Heterorhabditis and Steinernema are commercially used to control pest insects. They are symbiotically associated with bacteria of the genera Photorhabdus and Xenorhabdus, respectively, which are the major food source for the nematodes. The biology of the nematode-bacterium complex is described, a historical review of the development of in vitro cultivation techniques is given and the current use in agriculture is summarised. Cultures of the complex are pre-incubated with the symbiotic bacteria before the nematodes are inoculated. Whereas the inoculum preparation and preservation of bacterial stocks follow standard rules, nematodes need special treatment. Media development is mainly directed towards cost reduction, as the bacteria are able to metabolise a variety of protein sources to provide optimal conditions for nematode reproduction. The process technology is described, discussing the influence of bioreactor design and process parameters required to obtain high nematode yields. As two organisms are grown in one vessel and one of them is a multicellular organism, the population dynamics and symbiotic interactions need to be understood in order to improve process management. Major problems can originate from the delayed or slow development of the nematode inoculum and from phase variants of the symbiotic bacteria that have negative effects on nematode development and reproduction. Recent scientific progress has helped to understand the biological and technical parameters that influence the process, thus enabling transfer to an industrial scale. As a consequence, costs for nematode-based products could be significantly reduced.

Mass Production of the Beneficial Nematode Steinernema carpocapsae Utilizing a Fed-Batch Culturing Process

International Journal of Phytopathology

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 within a s...

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.

In vitro production of the biological control agent Heterorhabditis indica SL0708 in different agar media

Biocontrol Science and Technology, 2019

Heterorhabditis indica SL0708 is an entomopathogenic nematode isolated from Valle del Cauca-Colombia, whose bacterial symbiont, Photorhabdus luminescens subsp. akhurstii SL0708, has potential to control pests of economic importance in Colombia. Since in vivo production does not supply its demand, this investigation evaluated H. indica SL0708 production on different agar media. Five culture media (I, II, III, IV and V) were evaluated for productivity and pathogenicity of infective juveniles (IJs). IJs emerged between 11 and 16 days after inoculation in all media, with a total of 2.7 × 10 4 and 4.7 × 10 6 IJs produced during 15 days after IJs emergence, with maximum productivity at day five and high variability. Pathogenicity to Galleria mellonella larvae was not significantly different between in vitro and in vivo produced IJs on all media tested. Media IV and V were selected for their higher productivity. Subsequently, nematode inoculum size was evaluated in selected media at 2000, 4000 and 6000 IJs ml −1 , but significant differences were not observed in productivity and pathogenicity. Lastly, lipid source influence was evaluated in medium IV comparing canola, olive and soy oils. None of the plant-based oils had a significant effect on IJs production and pathogenicity. A medium was selected for H. indica SL0708 IJs production which was suitable in terms of productivity, culture time and pathogenicity of IJs produced. The medium and parameters selected in this study could be applied as an alternative for mass production of this entomopathogenic nematodes.

In vitro mass production of entomopathogenic nematodes on solid media: A review

Journal of entomology and zoology studies, 2020

Entomopathogenic nematodes, obligate insect pathogens, are widely used as biocontrol agents against economically important insect pests in different farming systems because they provide environmentally safe and sustainable crop protection. Hence the most important thing for the successful and reasonable usage of EPNs in crop protection is their production on large scale at competitive cost within a short time. In parallel to this, EPNs can be easily cultured either in-vivo or in vitro in the laboratory. The in vivo production is quite simple process as it involves the culturing on live insect host on the White Trap method, which involves the natural migration of IJs away from the infected host cadaver into the surrounding w+** ater layer. But the commercial production is quite impracticable due to high production costs, lacks of economies of scale and low nematode yields per gram of insect biomass. In place of that, the in vitro solid technology gives higher nematode yields per gram of solid media, which is based on introducing nematodes to a pure culture of the symbiotic bacteria into a nutritive, non-living medium that contains the sterile ingredients.

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.

eSci Journal of Plant Pathology MASS PRODUCTION OF THE BENEFICIAL NEMATODE STEINERNEMA CARPOCAPSAE UTILIZING A FED-BATCH CULTURING PROCESS A B S T R A C T

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...