The Efficacy of Entomopathogenic Nematodes Plus an Adjuvant against Helicoverpa zea and Chrysodeixis includens in Aboveground Applications.

In the southern United States, corn earworm, Helicoverpa zea (Boddie), and soybean looper, Chrysodeixis includens (Walker) are economically important crop pests. Although Bt crops initially provided effective control of target pests such as H. zea, many insect pests have developed resistance to these Bt crops. Alternative approaches are needed, including biological control agents such as entomopathogenic nematodes (EPNs). However, the effectiveness of EPNs for aboveground applications can be limited due to issues such as desiccation and ultraviolet radiation. Effective adjuvants are needed to overcome these problems. Ten strains of EPNs were tested for virulence against eggs, first to fourth instars, fifth instars, and pupae of H. zea and C. includens in the laboratory. These 10 EPN strains were Heterorhabditis bacteriophora (HP88 and VS strains), H. floridensis (K22 strain), Hgkesha (Kesha strain), Steinernema carpocapsae (All and Cxrd strains), S. feltiae (SN strain), S. rarum (17c+e strain), and S. riobrave (355 and 7-12 strains). EPNs could infect eggs of H. zea or C. includens in the laboratory, but the infection was low. The mortality caused by 10 EPN strains in seven days was significantly higher for the first to fourth instars of H. zea compared to the control, as was the fifth instars of H. zea. Similarly, for the first to fourth and fifth instars of C. includens, the mortality was significantly higher compared to the controls, respectively. However, only S. riobrave (355) had significantly higher mortality than the control for the pupae of H. zea. For the pupae of C. includens, except for H. bacteriophora (HP88), S. rarum (17c+e), and H. floridensis (K22), the mortality of the other seven strains was significantly higher than the control. Subsequently, S. carpocapsae (All) and S. riobrave (7-12) were chosen for efficacy testing in the field with an adjuvant 0.066% Southern Ag Surfactant (SAg Surfactant). In field experiments, the SAg Surfactant treatment significantly increased the mortality and EPN infection for S. carpocapsae (All) on first instars of H. zea in corn plant whorls. On soybean plants, with the SAg Surfactant, S. carpocapsae (All) was more effective than S. riobrave (7-12) on fifth instars of C. includens. This study indicates that EPNs can control H. zea and C. includens, and SAg Surfactant can enhance EPN efficacy.

Group Movement in Entomopathogenic Nematodes: Aggregation Levels Vary Based on Context.

Maintenance of an aggregated population structure implies within-species communication. In mixed-species environments, species-specific aggregations may reduce interspecific competition and promote coexistence. We studied whether movement and aggregation behavior of three entomopathogenic nematode species changed when isolated, as compared to mixed-species arenas. Movement and aggregation of Steinernema carpocapsae, S. feltiae and S. glaseri were assessed in sand. Each species demonstrated significant aggregation when alone. Mixed-species trials involved adding two species of nematodes, either combined in the center of the arena or at separate corners. While individual species became less aggregated than in single-species conditions when co-applied in the same location, they became more aggregated when applied in separate corners. This increased aggregation in separate-corner trials occurred even though the nematodes moved just as far when mixed together as they did when alone. These findings suggest that maintenance of multiple species within the same habitat is driven, at least in part, by species-specific signals that promote conspecific aggregation, and when the species are mixed (as occurs in some commercial formulations involving multiple EPN species), these signaling mechanisms are muddled.

Reproduction of Entomopathogenic Nematodes for Use in Pest Control.

The growing interest in the use of entomopathogenic nematodes and their symbiotic bacteria as promising biocontrol agents of many arthropod pests and pathogens has created running technologies to expand their use globally. The related laboratory procedures and tests on these nematodes such as their isolation, count, culture, identification, pathogenicity, virulence, and environmental tolerance should form the solid basis for such an expansion with reliable uses. Extensive practical details of such procedures and tests as well as how to identify and overcome the problems associated with these aspects are addressed in this chapter.

Enhancing entomopathogenic nematode efficacy with Pheromones: A field study targeting the pecan weevil.

Consistent efficacy is required for entomopathogenic nematodes to gain wider adoption as biocontrol agents. Recently, we demonstrated that when exposed to nematode pheromone blends, entomopathogenic nematodes showed increased dispersal, infectivity, and efficacy under laboratory and greenhouse conditions. Prior to this study, the impact of entomopathogenic nematode-pheromone combinations on field efficacy had yet to be studied. Steinernema feltiae is a commercially available entomopathogenic nematode that has been shown to increase mortality in insect pests such as the pecan weevil Curculio caryae. In this study, the pecan weevil was used as a model system to evaluate changes in S. feltiae efficacy when treated with a partially purified ascaroside pheromone blend. Following exposure to the pheromone blend, the efficacy of S. feltiae significantly increased as measured with decreased C. caryae survival despite unfavorable environmental conditions. The results of this study highlight a potential new avenue for using entomopathogenic nematodes in field conditions. With increased efficacy, using entomopathogenic nematodes will reduce reliance on conventional management methods in pecan production, translating into more environmentally acceptable practices.

STEINERNEMA ADAMSI N. SP. (RHABDITIDA: STEINERNEMATIDAE), A NEW ENTOMOPATHOGENIC NEMATODE FROM THAILAND.

A new species of entomopathogenic nematode, Steinernema adamsi n. sp., was recovered from the soil of a longan tree (Dimocarpus sp.) in Mueang Lamphun District, Thailand, using baiting techniques. Upon analysis of the nematode's morphological traits, we found it to be a new species of Steinernema and a member of the Longicaudatum clade. Molecular analyses of the ITS rDNA and D2D3 of 28S rDNA sequences further confirmed that S. adamsi n. sp. is a new species of the Longicaudatum clade, which is closely related to Steinernema guangdongense and Steinernema longicaudam. Using morphometric analysis, the infective juveniles measure between 774.69 and 956.96 µm, males have a size range of 905.44 to 1,281.98 µm, and females are within the range of 1,628.21 to 2,803.64 µm. We also identified the symbiotic bacteria associated with the nematode based on 16S sequences as Xenorhabdus spp. closely related toXenorhabdus griffiniae. Furthermore, we have successfully assessed a cryopreservation method for the long-term preservation of S. adamsi n. sp. Successful cryopreservation of this new species will allow for the longer preservation of its traits and will be valuable for its future use. The discovery of this new species has significant implications for the development of effective biological control agents in Thailand, and our work contributes to our understanding of the diversity and evolution of entomopathogenic nematodes.

Antifungal Effect of Metabolites from Bacterial Symbionts of Entomopathogenic Nematodes on Fusarium Head Blight of Wheat.

Fungal diseases such as Fusarium head blight (FHB) are significant biotic stressors, negatively affecting wheat production and quality. This study explored the antifungal activity of the metabolites produced by the bacterial symbionts of entomopathogenic nematodes (EPNs) against FHB-causing Fusarium sp. Fusarium graminearum. To achieve this, the symbiotic bacteria of nine EPN isolates from the EPN collection at the Agricultural Research Council-Small Grains (ARC-SG) were isolated from the cadavers of Galleria mellonella (Lepidoptera: Pyralidae) larvae after infection with EPNs. Broth cultures (crude) and their supernatants (filtered and autoclaved) of each bacterial isolate were used as bacterial metabolite treatments to test their inhibitory effect on the mycelial growth and spore germination of F. graminearum. Mycelial growth inhibition rates varied among both bacterial isolates and treatments. Crude metabolite treatments proved to be more effective than filtered and autoclaved metabolite treatments, with an overall inhibition rate of 75.25% compared to 23.93% and 13.32%, respectively. From the crude metabolite treatments, the Xenorhabdus khoisanae SGI 197 bacterial isolate from Steinernema beitlechemi SGI 197 had the highest mean inhibition rate of 96.25%, followed by Photorhabdus luminescens SGI 170 bacteria isolated from Heterorhabditis bacteriophora SGI 170 with a 95.79% mean inhibition rate. The filtered metabolite treatments of all bacterial isolates were tested for their inhibitory activity against Fusarium graminearum spore germination. Mean spore germination inhibition rates from Xenorhabdus spp. bacterial isolates were higher (83.91 to 96.29%) than those from Photorhabdus spp. (6.05 to 14.74%). The results obtained from this study suggest that EPN symbiotic bacterial metabolites have potential use as biological control agents of FHB. Although field efficacy against FHB was not studied, the significant inhibition of mycelial growth and spore germination suggest that the application of these metabolites at the flowering stage may provide protection to plants against infection with or spread of F. graminearum. These metabolites have the potential to be employed as part of integrated pest management (IPM) to inhibit/delay conidia germination until the anthesis (flowering stage) of wheat seedlings has passed.

Functional Comparison of Three Chitinases from Symbiotic Bacteria of Entomopathogenic Nematodes.

Xenorhabdus and Photorhabdus, bacterial symbionts of entomopathogenic nematodes Steinernema and Heterorhabditis, respectively, have several biological activities including insecticidal and antimicrobial activities. Thus, XnChi, XhChi, and PtChi, chitinases of X. nematophila, X. hominickii, and P. temperata isolated from Korean indigenous EPNs S. carpocapsae GJ1-2, S. monticolum GJ11-1, and H. megidis GJ1-2 were cloned and expressed in Escherichia coli BL21 to compare their biological activities. Chitinase proteins of these bacterial symbionts purified using the Ni-NTA system showed different chitobiosidase and endochitinase activities, but N-acetylglucosamidinase activities were not shown in the measuring of chitinolytic activity through N-acetyl-D-glucosarmine oligomers. In addition, the proteins showed different insecticidal and antifungal activities. XnChi showed the highest insecticidal activity against Galleria mellonella, followed by PtChi and XhChi. In antifungal activity, XhChi showed the highest half-maximal inhibitory concentration (IC50) against Fusarium oxysporum with 0.031 mg/mL, followed by PtChi with 0.046 mg/mL, and XnChi with 0.072 mg/mL. XhChi also showed the highest IC50 against F. graminearum with 0.040 mg/mL, but XnChi was more toxic than PtChi with 0.055 mg/mL and 0.133 mg/mL, respectively. This study provides an innovative approach to the biological control of insect pests and fungal diseases of plants with the biological activity of symbiotic bacterial chitinases of entomopathogenic nematodes.

Molecular identification of a peroxidase gene controlling body size in the entomopathogenic nematode Steinernema hermaphroditum.

The entomopathogenic nematode Steinernema hermaphroditum was recently rediscovered and is being developed as a genetically tractable experimental system for the study of previously unexplored biology, including parasitism of its insect hosts and mutualism with its bacterial endosymbiont Xenorhabdus griffiniae. Through whole-genome re-sequencing and genetic mapping we have for the first time molecularly identified the gene responsible for a mutationally defined phenotypic locus in an entomopathogenic nematode. In the process we observed an unexpected mutational spectrum following ethyl methansulfonate mutagenesis in this species. We find that the ortholog of the essential Caenorhabditis elegans peroxidase gene skpo-2 controls body size and shape in S. hermaphroditum. We confirmed this identification by generating additional loss-of-function mutations in the gene using CRISPR-Cas9. We propose that the identification of skpo-2 will accelerate gene targeting in other Steinernema entomopathogenic nematodes used commercially in pest control, as skpo-2 is X-linked and males hemizygous for loss of its function can mate, making skpo-2 an easily recognized and maintained marker for use in co-CRISPR.

Antimicrobial Peptides (AMP) in the Cell-Free Culture Media of Xenorhabdus budapestensis and X. szentirmaii Exert Anti-Protist Activity against Eukaryotic Vertebrate Pathogens including Histomonas meleagridis and Leishmania donovani Species.

Anti-microbial peptides provide a powerful toolkit for combating multidrug resistance. Combating eukaryotic pathogens is complicated because the intracellular drug targets in the eukaryotic pathogen are frequently homologs of cellular structures of vital importance in the host organism. The entomopathogenic bacteria (EPB), symbionts of entomopathogenic-nematode species, release a series of non-ribosomal templated anti-microbial peptides. Some may be potential drug candidates. The ability of an entomopathogenic-nematode/entomopathogenic bacterium symbiotic complex to survive in a given polyxenic milieu is a coevolutionary product. This explains that those gene complexes that are responsible for the biosynthesis of different non-ribosomal templated anti-microbial protective peptides (including those that are potently capable of inactivating the protist mammalian pathogen Leishmania donovanii and the gallinaceous bird pathogen Histomonas meleagridis) are co-regulated. Our approach is based on comparative anti-microbial bioassays of the culture media of the wild-type and regulatory mutant strains. We concluded that Xenorhabdus budapestensis and X. szentirmaii are excellent sources of non-ribosomal templated anti-microbial peptides that are efficient antagonists of the mentioned pathogens. Data on selective cytotoxicity of different cell-free culture media encourage us to forecast that the recently discovered "easy-PACId" research strategy is suitable for constructing entomopathogenic-bacterium (EPB) strains producing and releasing single, harmless, non-ribosomal templated anti-microbial peptides with considerable drug, (probiotic)-candidate potential.

Positive Chemotaxis of the Entomopathogenic Nematode Steinernema australe (Panagrolaimorpha: Steinenematidae) towards High-Bush Blueberry (Vaccinium corymbosum) Root Volatiles.

The foraging behavior of the infective juveniles (IJs) of entomopathogenic nematodes (EPNs) relies on host-derived compounds, but in a tri-trophic context, herbivore-induced root volatiles act as signals enhancing the biological control of insect pests by recruiting EPNs. In southern Chile, the EPN Steinernema australe exhibits the potential to control the raspberry weevil, Aegorhinus superciliosus, a key pest of blueberry Vaccinium corymbosum. However, there is no information on the quality of the blueberry root volatile plume or the S. australe response to these chemicals as putative attractants. Here, we describe the root volatile profile of blueberries and the chemotaxis behavior of S. australe towards the volatiles identified from Vaccinium corymbosum roots, infested or uninfested with A. superciliosus larvae. Among others, we found linalool, α-terpineol, limonene, eucalyptol, 2-carene, 1-nonine, 10-undecyn-1-ol, and methyl salicylate in root volatiles and, depending on the level of the emissions, they were selected for bioassays. In the dose-response tests, S. australe was attracted to all five tested concentrations of methyl salicylate, 1-nonine, α-terpineol, and 2-carene, as well as to 100 µg mL-1 of 10-undecyn-1-ol, 0.1 and 100 µg mL-1 of linalool, and 100 µg mL-1 of limonene, whereas eucalyptol elicited no attraction or repellency. These results suggest that some volatiles released from damaged roots attract S. australe and may have implications for the biocontrol of subterranean pests.

The Coincidental Evolution of Virulence Partially Explains the Virulence in a Generalist Entomopathogenic.

PURPOSE: The parasites' virulence is labile after jumping to a new host species, and it might derivate in gaining virulence against a new host as a side effect of living in a non-host environment (coincidental evolution of virulence hypothesis). METHODS: To test this hypothesis, we monitored the experimental evolution of the Rhabditis regina nematode for over 290 generations (4 years) in three environments (strains): (1) the natural host, Phyllophaga polyphylla, (2) an alternate host, Tenebrio molitor, and (3) saprophytic medium (beef; the food that may provide evidence for the coincidental evolution of virulence). Each strain was exposed to P. polyphylla, T. molitor, or Galleria mellonella. We compared the host survival and immune response (proPO, PO, and lytic activity) of infected versus uninfected hosts. RESULTS: The saprophytic nematodes gained virulence only against G. mellonella. However, the P. polyphylla strain was more effective in killing P. polyphylla than T. molitor, and the T. molitor strain was more effective against T. molitor than P. polyphylla. Additionally, one dauer larva was sufficient to kill the hosts. Finally, the immune response did not differ between the challenged and control groups. CONCLUSION: The coincidental evolution of virulence partially explains our results, but they might also support the short-sighted hypothesis. Additionally, we found evidence for immunomodulation because nematodes passed unnoticed to the immune response. It is crucial to analyze the virulence of entomopathogens from the point of view of the evolution of virulence to be aware of potential scenarios that might limit biological control.

PI3K/Akt/CncC signaling pathway mediates the response to EPN-Bt infection in Holotrichia parallela larvae.

BACKGROUND: Combining the entomopathogenic nematode (EPN), Heterorhabditis beicherriana LF strain, and Bacillus thuringiensis (Bt) HBF-18 strain is a practical strategy to manage the larvae of Holotrichia parallela Motschulsky (white grubs). However, the mechanisms underlying the larval defense response to this combined biocontrol strategy are unknown. RESULTS: The activities of some antioxidant enzymes (SOD, POD, CAT) and some detoxifying enzymes (AChE, P-450, CarE, GST) in grubs showed an activation-inhibition trend throughout the EPN-Bt exposure time course. Eight potentially key antioxidant and detoxifying enzyme genes in response to EPN-Bt infection were identified from the midgut of grubs through RNA sequencing. After silencing CAT, CarE18, and GSTs1, the enzyme activities were significantly decreased by 30.29%, 68.80%, and 34.63%, respectively. Meanwhile, the mortality of grubs was increased by 18.40%, 46.30%, and 42.59% after exposure to EPN-Bt for 1 day. Interestingly, the PI3K/Akt signaling pathway was significantly enriched in KEGG enrichment analysis, and the expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), cap 'n' collar isoform-C (CncC), kelch-like ECH-associated protein 1 (Keap1), and CarE18 were all up-regulated when exposed to EPN-Bt for 1 day. Furthermore, RNAi-mediated PI3K silencing showed a similar down-regulated trend between PI3K/Akt/CncC and CarE18. Moreover, silencing PI3K rendered grubs more susceptible to EPN-Bt and accelerated symbiotic bacteria multiplication in grubs. CONCLUSION: These results suggest that the PI3K/Akt/CncC pathway mediates the expression of CarE18 and participates in the defense response of H. parallela larvae against EPN-Bt infection. Our data provide valuable insights into the design of appropriate management strategies for this well-known agricultural pest. © 2022 Society of Chemical Industry.

Tolerance of Steinernema carpocapsae infective juveniles in novel nanoparticle formulations to ultraviolet radiation.

Entomopathogenic nematodes (EPNs) are susceptible to abiotic environmental factors including ultraviolet (UV) radiation, which affects the survival and efficacy. This study evaluated nanoparticle (NP) formulations for protecting Steinernema carpocapsae infective juveniles (IJs) from UV radiation. First, silica-NH2 NPs at oil-to-water ratios of 2:8, 3:7 and 4:6 were compared with Barricade Fire Gel (1 % and 2 %) and a water control (aqueous IJs) by exposing IJs to UV light (254 nm) for 0, 10 and 20 min. Barricade gel (especially 2 % Barricade) significantly improved IJs viability after UV treatment, while all three NPs had adverse effects on IJ viability after UV radiation. Subsequently, two silica (SiO2 basic and advanced) and one titania (TiO2) based formulations were tested with Barricade (1 % and 2 %) and a water control. The titania-NH2 NPs provided the highest UV protection, and IJ viability and virulence were not reduced even after 20-min UV. Except TiO2, only 2 % Barricade at 10-min UV and SiO2 basic at 20-min UV had lower IJ mortality than the water control. Only TiO2 formulated IJs caused higher insect mortality and infection levels than aqueous IJs after UV treatment. The UV tolerance of TiO2 was further examined by assessing the number of nematodes invading the hosts. Consistent with virulence tests, the number of invading nematodes in titania-NH2 NPs did not decrease after UV radiation for 10 or 20 min compared with the no-UV control. The anti-UV capability of titania-NH2 NPs has promise as a tool to enhance biocontrol efficacy of EPNs under field conditions.

A novel volatile deterrent from symbiotic bacteria of entomopathogenic nematodes fortifies field performances of nematodes against fall armyworm larvae.

The core elements of entomopathogenic nematode toxicity towards the fall armyworm Spodoptera frugiperda are associated with symbiotic bacteria. These microbes provide independent control effects and are reported to have repellency to insect pests. However, the ecological background of this nematode-bacteria-insect communication module is elusive. This work aims to identify key chemical cues which drive the trophic interactions through olfactory reception of S. frugiperda, and to inspire implementations with these isolated behavioral regulators in the corn field. A total of 657 volatiles were found within 13 symbiotic bacterial strains, and five of them induced significant electrophysiological responses of S. frugiperda larvae. 2-Hexynoic acid was demonstrated to exhibit a dominant role in deterring S. frugiperda larvae from feeding and localization. Field implementations with this novel volatile deterrent have resulted in fortified nematode applications. 2-Hexynoic acid acts as an excellent novel deterrent and presents remarkable application potential against fall armyworm larvae. Emissions from symbiotic bacteria of entomopathogenic nematodes are key players in chemical communication among insects, nematodes, and microbes. The olfactory perceptions and molecular targets for this volatile are worthy of future research.

Entomopathogenic Nematodes and Their Symbiotic Bacteria from the National Parks of Thailand and Larvicidal Property of Symbiotic Bacteria against Aedes aegypti and Culex quinquefasciatus.

Entomopathogenic nematodes (EPNs) are insect parasitic nematodes of the genera Het-erorhabditis and Steinernema. These nematodes are symbiotically associated with the bacteria, Photorhabdus and Xenorhabdus, respectively. National parks in Thailand are a potentially rich resource for recovering native EPNs and their symbiotic bacteria. The objectives of this study are to isolate and identify EPNs and their bacterial flora from soil samples in four national parks in Thailand and to evaluate their efficacy for controlling mosquito larvae. Using a baiting method with a Galleria mellonella moth larvae and a White trap technique, 80 out of 840 soil samples (9.5%) from 168 field sites were positive for EPNs. Sequencing of an internal transcribed spacer resulted in the molecular identification of Heterorhabditis nematode isolates as H. indica, H. baujardi and Heterorhabditis SGmg3, while using 28S rDNA sequencing, Steinernema nematode species were identified as S. guang-dongense, S. surkhetense, S. minutum, S. longicaudum and one closely related to S. yirgalemense. For the symbiotic bacterial isolates, based on recA sequencing, the Photorhabdus spp. were identified as P. luminescens subsp. akhurstii, P. luminescens subsp. hainanensis and P. luminescens subsp. australis. Xenorhabdus isolates were identified as X. stockiae, X. indica, X. griffiniae, X. japonica and X. hominickii. Results of bioassays demonstrate that Photorhabdus isolates were effective on both Aedes aegypti and Culex quinquefasciatus. Therefore, we conclude that soil from Thailand's national parks contain a high diversity of entomopathogenic nematodes and their symbiotic bacteria. Photorhabdus bacteria are larvicidal against culicine mosquitoes and may serve as effective biocontrol agents.

Nematophilic bacteria associated with entomopathogenic nematodes and drug development of their biomolecules.

The importance of Xenorhabdus and Photorhabdus symbionts to their respective Steinernema and Heterorhabditis nematode hosts is that they not only contribute to their entomopathogenicity but also to their fecundity through the production of small molecules. Thus, this mini-review gives a brief introductory overview of these nematophilic bacteria. Specifically, their type species, nematode hosts, and geographic region of isolations are tabulated. The use of nucleotide sequence-based techniques for their species delineation and how pangenomes can improve this are highlighted. Using the Steinernema-Xenorhabdus association as an example, the bacterium-nematode lifecycle is visualized with an emphasis on the role of bacterial biomolecules. Those currently in drug development are discussed, and two potential antimalarial lead compounds are highlighted. Thus, this mini-review tabulates forty-eight significant nematophilic bacteria and visualizes the ecological importance of their biomolecules. It further discusses three of these biomolecules that are currently in drug development. Through it, one is introduced to Xenorhabdus and Photorhabdus bacteria, their natural production of biomolecules in the nematode-bacterium lifecycle, and how these molecules are useful in developing novel therapies.

Toxicity of Photorhabdus luminescens and Xenorhabdus bovienii bacterial metabolites to pecan aphids (Hemiptera: Aphididae) and the lady beetle Harmonia axyridis (Coleoptera: Coccinellidae).

Toxicity of the metabolites of two bacteria, Photorhabdus luminescens and Xenorhabdus bovienii, symbionts of entomopathogenic nematodes, were tested in the laboratory against the multicolored Asian lady beetle, Harmonia axyridis, the black pecan aphid, Melanocallis caryaefoliae, and the blackmargined aphid, Monellia caryella. Bacterial broth prepared from both P. luminescens and X. bovienii demonstrated high levels of toxicity equivalent to the pyrethroid insecticide bifenthrin and caused higher insect mortality than tryptic soy broth plus yeast extract (TSY) (blank control) against M. caryella; broth culture of P. luminescens was more effective than TSY against M. caryaefoliae. At the levels tested, the metabolites were not toxic to H. axyridis.

Modeling the potential global distribution of suitable habitat for the biological control agent Heterorhabditis indica.

Entomopathogenic nematode (EPN) Heterorhabditis indica is a promising biocontrol candidate. Despite the acknowledged importance of EPN in pest control, no extensive data sets or maps have been developed on their distribution at global level. This study is the first attempt to generate Ecological Niche Models (ENM) for H. indica and its global Habitat Suitability Map (HSM) for H. indica to generate biogeographical information and predicts its global geographical range and help identify of prospective areas for its exploration and to help identify the suitable release areas for biocontrol purpose. The aim of the modeling exercise was to access the influence of temperature and soil moisture on the biogeographical patterns of H. indica at the global level. Global Heterorhabditis indica ecosystems. CLIMEX software was used to model the distribution of H. indica and assess the influence of environmental variable on its global distribution. In total, 162 records of H. indica occurrence from 27 countries over 25 years were combined to generate the known distribution data. The model was further fine-tuned using the direct experimental observations of the H. indica's growth response to temperature and soil moisture. Model predicts that much of the tropics and subtropics have suitable climatic conditions for H. indica. It further predicts that H. indica distribution can extend into warmer temperate climates. Examination of the model output, predictions maps at a global level indicate that H. indica distribution may be limited by cold stress, heat stress, and dry stresses in different areas. However, cold stress appears to be the major limiting factor. This study highlighted an efficient way to construct HSM for EPN potentially useful in the search/release of target species in new locations. The study showed that H. indica which is known as warm adapted EPN generally found in tropics and subtropics can potentially establish itself in warmer temperate climates as well. The model can also be used to decide the release timing of EPN by adjusting with season for maximum growth. The model developed in this study clearly identified the value and potential of Habitat Suitability Map (HSM) in planning of future surveys and application of H. indica.

Dimethyl sulfoxide and ascarosides improve the growth and yields of entomopathogenic nematodes in liquid cultures.

Increasing the infective juvenile (IJ) yields of entomopathogenic nematodes in monoxenic culture systems would reduce their production cost for the market. Ascarosides act as universal nematode pheromones with developmental and behavioral effects of nematodes. Dimethyl sulfoxide (DMSO) is unexpectedly found to enhance the IJ yields of entomopathogenic nematodes on fortified nutrient broth plates. In this study, the influence of selected ascarosides (ascr#7, ascr#9 and ascr#11) and DMSO in three concentrations on the IJ yields of S. carpocapsae All and H. bacteriophora H06 in liquid culture flasks was determined, and the critical development parameters (IJ recovery rate, number of hermaphrodites, number of visible eggs in a hermaphrodite) were examined for H. bacteriophora H06. The results demonstrated that IJ yields were significantly improved in the liquid medium containing 0.01 % DMSO, and 0.02 nM ascr#11 for S. carpocapsae All, and 0.1 % and 0.01 % DMSO and 0.02 pM ascr#11 for H. bacteriophora H06 in proper concentrations. Furthermore, it was discovered that increased recovery rate, hermaphrodite numbers and eggs in the hermaphrodites may contribute to the improved IJ yields of H. bacteriophora H06 in DMSO-supplemented liquid medium. Compared with the control flasks, the IJ yields from the flasks containing 0.01 % DMSO were 15 % and 35 % higher for S. carpocapsae All and H. bacteriophora H06 respectively in 15 days. The cost for ascarosides and DMSO is almost negligible. The results would provide practical technology for low-cost commercial production of these nematodes for pest management program.