Automated technology for in vivo mass production of entomopathogenic nematodes (original) (raw)
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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.
New Approaches For Extracting And Mass Rearing of Entomopathogenic Nematodes In Vivo
Egyptian Academic Journal of Biological Sciences, B. Zoology
Entomopathongic nematodes (EPNs) including stienernematids and heterorhabditis are used to control a variety of economically insect pests (Shapiro-Ilan et al., 2002b; Grewal et al., 2005). These nematodes gave control equivalent to or superior than the chemical standard insecticides. The infective juveniles (IJs) enter their host through natural openings or through the cuticle. Subsequently, nematodes release their bacterial symbioses, which are responsible for killing the host within 24-48 hours (Dowds and Peters, 2002). The nematodes feed on the bacterial cells and host tissues that has been metabolized by the bacterium and has 1-3 generations, depending ARTICLE INFO ABSTRACT Article History
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- 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.
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 multiplication of entomopathogenic nematodes in artificial media
Journal of entomology and zoology studies, 2019
This study evaluated ten different media of plant origin and four different media of animal origin for in vitro mass production of Heterorhabditis bacteriophora, Steinernema aciari and Oscheius chongmingensis. Maximum multiplication of all the tested species was observed in the media of plant origin containing green gram and coconut oil. Maximum multiplication of all the tested species was observed in the media of animal origin supplemented with pork waste homogenate.
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.
Biological Control, 2006
A wide range of technology is available for application of entomopathogenic nematodes including various irrigation systems and spray equipment. The choice of application equipment, and manner in which the nematodes are applied, can have substantial impact on pest control eYcacy. For example, nozzle or pumping system types are some of the parameters that can aVect nematode performance following spray applications. Operating pressures for some nematode species may reach up to 2000 kPa without notable damage, whereas other species may require lower pressure limits, e.g., 1380 kPa for Heterorhabditis megidis. In addition to application equipment, a variety of other abiotic and biotic factors must be considered. In general, a rate of 25 infective juvenile nematodes/cm 2 is required for successful pest suppression. Critical environmental factors include avoidance of ultraviolet radiation, adequate soil moisture, and appropriate temperature. Certain fertilizers and chemical pesticides can have positive eVects on entomopathogenic nematode eYcacy, whereas other agents may have neutral or negative eVects. Similarly, certain biotic agents present during soil applications can be expected to be detrimental to nematode applications (e.g., nematophagous mites and fungi), whereas other organisms may be beneWcial (e.g., some combinations with Bacillus thuringiensis). With some exceptions foliar applications have been less successful than soil applications due to nematode susceptibility to desiccation and UV; recent research, however, indicates that frequent low-rate applications of nematodes to foliage can result in substantial suppression of greenhouse pests such as thrips. Further innovation in application technology will undoubtedly contribute to the expansion of entomopathogenic nematodes as biocontrol agents. Published by Elsevier Inc.
TURKISH JOURNAL OF AGRICULTURE AND FORESTRY, 2015
Introduction Entomopathogenic nematodes (EPNs) (Steinernematidae and Heterorhabditidae) are mutualistically associated with insect-pathogenic bacteria and together they kill their insect hosts. The steinernematids are associated with the bacterial genus Xenorhabdus, whereas the heterorhabditids are associated with the genus Photorhabdus. These EPNs have adapted specific mechanisms to transmit the bacteria to their insect hosts (Dillman et al., 2012) and are considered good candidates for integrated pest management of soil insect pests (Lacey and Georgis, 2012). In fact, several nematode species are produced commercially and applied in a variety of cropping systems in many different countries (Alves, 1986; Garcia et al., 2008). These biological control agents must be delivered in a way that enables the infective juveniles (IJs) of the nematodes to survive and infect their hosts (Shapiro-Ilan et al., 2006; Brusselman et al., 2012). EPNs can be applied with nearly all agronomic or horticultural ground equipment including pressurized tank sprayers, mist blowers, electrostatic sprayers, drip irrigation systems, or even aerial sprayers (Georgis, 1990;
Biosystematics of entomopathogenic nematodes: current status, protocols and definitions
Journal of Helminthology, 1997
COST Action 819:Entomopathogenic nematodes, supercedes Action 812:Cold active lines of insect parasitic nematodes in Agriculture and Biotechnology. It functions in the field of Agriculture and Biotechnology and began in July 1994 and will end in May, 1999. The main objective is to combine interrelated European expertise to increase the use of entomopathogenic nematodes (EPNs) in integrated pest management and to reduce the need for chemical control. Coordination of the Action is the responsibility of a management committee in accordance with a Memorandum of Understanding, which has been signed by representatives of 17 countries: Austria, Belgium, Czech Republic, Finland, France, Germany, Hungary, Ireland, Italy, The Netherlands, Norway, Poland, Portugal, Spain, Sweden, Switzerland and the UK. A research institute in Israel is also participating. Over 40 research institutions and 10 commercial companies participate in Action 819. The EU funding meets the costs related to coordination...