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Open access peer-reviewed chapter

Entomopathogenic Nematode: A Sustainable Option for IPM

Written By

Thattankandy Meethal Aparna, Mukesh R. Siddhapara and Sonam Kumari

Submitted: 19 April 2025 Reviewed: 06 May 2025 Published: 25 July 2025

DOI: 10.5772/intechopen.1010919

Nematodes IntechOpen
Nematodes From the Past to the Future Edited by Nihal Dogan

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Nematodes - From the Past to the Future

Nihal Dogan

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Abstract

The chapter provides a comprehensive overview of entomopathogenic nematodes (EPNs) as viable biocontrol agents in biointensive pest management systems, particularly in the context of sustainable agriculture. Despite the underdeveloped biopesticide market, the capabilities of entomopathogenic nematodes to combat various insect pests, including those in the orders Coleoptera, Diptera, Lepidoptera, and Hemiptera, are highlighted. The chapter discusses the historical context of entomopathogenic nematode discovery, their symbiotic relationships with bacteria, and the methodologies for their isolation and mass production, including both in vivo and in vitro methods. The efficacy of various entomopathogenic nematode species against target pests is substantiated through laboratory studies and field, demonstrating significant mortality rates across multiple pest species. Additionally, the functional compatibility of entomopathogenic nematodes with chemical pesticides and entomopathogens is examined, suggesting their integrative role in pest management strategies. The findings underscore the ecological benefits of using entomopathogenic nematodes as a sustainable alternative to synthetic pesticides, emphasizing their safety for non-target organisms and humans. Overall, the study advocates for the integration of entomopathogenic nematodes into integrated pest management practices to boost pest control efficacy and promote sustainable agricultural practices in India.

Keywords

  • in vivo and in vitro multiplication
  • EPN
  • mass production
  • symbiotic bacteria
  • infective juveniles
  • IPM
  • mortality
  • biological control

1. Introduction

Despite initial expectations, the biopesticide sector in India has experienced limited growth and continues to occupy a relatively smaller share compared to the synthetic pesticide market. This slower development is attributed to various industrial and policy-related obstacles that have constrained production capacity. However, efforts to promote biopesticides as part of sustainable agricultural practices have been reinforced by initiatives such as the National Farmer Policy of 2007 [1].

The biopesticides market was valued at approximately USD 7.3 billion in 2023 and is projected to grow from USD 8.32 billion in 2024 to USD 23.74 billion by 2032, exhibiting a compound annual growth rate (CAGR) of 14.0% during the forecast period (2025–2032). As of November 2024, a total of 970 biopesticides have been registered with the Central Insecticides Board and Registration Committee (CIBRC) under the Government of India [2]. It is estimated that by 2050, biopesticides will constitute up to 50% of the Indian pesticide market [1], a significant increase from their current share of approximately 9% of total pesticide consumption [3]. An annual growth rate of 2.5% is anticipated for the sector. Nevertheless, at present, the biopesticide market remains underdeveloped relative to initial expectations and continues to represent a modest segment compared to the synthetic pesticide industry. Production remains comparatively limited, largely due to persistent challenges at both industrial and policy levels.

Scientific evidence shows that in modern industrial agriculture, the use of synthetic pesticides contributes significantly to greenhouse gas emissions (US Environment Protection Agency 2022 [4]) and also makes our agricultural systems more vulnerable to the effects of climate change [5, 6]. Pesticides contribute to climate change throughout their entire life cycle, through production, packaging, transportation, application, and even environmental degradation and disposal [7].

Integrated Pest Management (IPM) is an ecosystem-based approach to pest control that combines multiple practices to maintain pest populations below economic threshold levels. In recent years, the incorporation of entomopathogenic nematodes (EPNs) as a component of IPM strategies has gained increasing attention, primarily to mitigate issues such as insecticide resistance, pest resurgence, and pesticide residue accumulation. Entomopathogenic nematodes (derived from ‘entomo’ meaning insect, and ‘pathogenic’ meaning disease-causing) are microscopic, soil-dwelling roundworms capable of infecting and killing insect pests. These nematodes utilize symbiotic bacteria to cause lethal infections in their hosts, making them effective biological control agents within sustainable pest management frameworks.

EPNs are exempt from registration and regulatory requirements by the United States Environmental Protection Agency (EPA), the Central Insecticides Board and Registration Committee (CIBRC) of India, and analogous regulatory bodies in several other countries.

Various biocontrol applications have used entomopathogenic nematodes as control agent [8], with inundative biological control forming the largest [9, 10]. In inundative biological control, large numbers of nematodes are released to invade the pest population and achieve immediate pest suppression [9]. Such control measures have been successfully used in field treatments for a range of entomological pests, including the insect orders Coleoptera, Diptera, Orthoptera, Hemiptera, and Lepidoptera [11, 12, 13].

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2. History of EPN

The first entomopathogenic nematode (EPN) was initially described by Steiner in 1923 as Aplectana kraussei, which is now classified as Steinernema kraussei. In 1929, Steiner also introduced Neoaplectana glaseri, the first EPN species to be utilized for biological control, specifically targeting white grubs. In 1955, a nematode species, Neoaplectana carpocapsae, was identified by Weiser from the larvae of the codling moth (Carpocapsa pomonella). Interestingly, Dutky and Hough independently discovered the same species in the same host during the same year, with the isolate being designated as DD-136 for use in pest control. Heterorhabditis bacteriophora was first described by Pionar in 1976. Later, in 1994, Nguyen and Smart proposed the genus Neosteinernema, containing the species N. longicurvicauda. The most widely studied and utilized EPN species are found within the genera Steinernema and Heterorhabditis, which encompass the most significant species for biological control applications [14].

Entomopathogenic nematodes (EPNs) have developed the ability to detect and respond to plant-derived signals in order to locate potential herbivore hosts. When herbivores infest plant roots, they release a distinct set of molecules that differentiate them from healthy plants. The presence and spread of herbivore-induced plant volatiles (HIPVs) serve as a reliable indicator of herbivore activity. Additionally, EPNs utilize carbon dioxide, which is emitted by most living organisms, as a key cue for host location. This signal prompts foraging behavior in EPNs, with different life strategies such as ambushers and cruisers exhibiting varying levels of sensitivity to this stimulus. Furthermore, EPNs can exploit insect-specific chemical cues to locate their hosts more effectively. Herbivores continuously release various chemicals into the surrounding environment, including pheromones, exudates, molted skins (exuviae), and feces (frass). These insect-derived compounds, diffusing through the soil, can either attract or repel EPNs, or even induce behaviors such as nictation [15].

Heterorabditidae and Steinernematide are commonly used genera in pest management programs even though there are about 40 nematode families reported to control insect pests [16]. Around 100 species of Steinernema and 26 species of Heterorhabditis have been recorded worldwide [17, 18]. Steinernema and Heterorhabditis EPNs are known to be parasites of lepidopterans, coleopterans, and occasionally of orthopterans, dipterans, and hymenopterans [19, 20, 21].

Entomopathogenic nematodes (EPNs) employ two primary foraging strategies: ambushers and cruisers. Ambushers, such as Steinernema carpocapsae, adopt an energy-conserving approach, remaining stationary and waiting for mobile insect hosts to come within their reach, typically targeting pests in the upper soil layers like armyworms and cutworms. In contrast, cruisers are more active and typically subterranean, capable of traveling considerable distances to locate their host. These nematodes, including Steinernema glaseri and Heterorhabditis bacteriophora, are particularly effective against less mobile soil-dwelling pests, such as white grubs (Scarab beetles).

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3. White trap method and multiplication

Infective juveniles (IJs) of entomopathogenic nematodes (EPNs) are commonly found in the soil and can be readily harvested by collecting samples from diverse environments such as agricultural fields, forests, riverbanks, pastures, and orchards across varying climatic conditions. Sampling is typically conducted using tools such as augers, tubes, shovels, and trowels. Two primary sampling techniques are generally used: stratified sampling and random sampling. Soil samples are typically collected from depths of up to 25 cm and from areas approximately 4 to 5 m2 in size. A set of 3 to 5 sub-samples are taken per sample site and combined into a plastic bag for analysis. It is essential that the collected samples are clearly labeled with detailed information, including the location or site (preferably with GPS coordinates), the date of sampling, habitat type, prevailing vegetation, climatic conditions, altitude, and any insect pests observed at the sampling site.

The likelihood of naturally encountering EPN-infected insect cadavers in the field is relatively low, making insect baiting an essential and efficient method for isolating entomopathogenic nematodes (EPNs) from soil samples. Insect baiting is considered one of the simplest and most cost-effective approaches for attracting EPNs as well as insect pathogenic fungi [22]. In this technique, up to 100 grams of soil are placed in clean containers, and 4 to 5 Galleria larvae are introduced as bait. The containers are then sealed, inverted, and incubated in the dark at 25°C for 2 to 3 days, during which time mortality is monitored. Infected larvae exhibit distinct color changes: Steinernema-infected larvae turn yellow to brown, while Heterorhabditis-infected larvae turn red (Figure 1A, B). The cadavers are then carefully rinsed in sterile water and placed in white traps for the subsequent extraction of infective juveniles (IJs) and other developmental stages [24].

Figure 1.

Methodology for mass multiplication of Entomopathogenic nematode. Cadavers with body turned to a yellow to brown color indicate infection with Steinernematids, while those that become red are infected by Heterorhabditids (A, B). The deceased cadavers are placed on the small petri dish, a small amount of sterile water is added, Fresh IJs and other developmental stages can be collected from the suspensions after a duration of 10 to 15 days (C, D) [4]. (Source: Singh et al. [23]).

The white trap consists of a large dish (100 mm in diameter) containing a smaller petri dish (50 mm in diameter) lined with filter paper. Infected cadavers are placed on the smaller petri dish, and a small volume of sterile water is added. The setup is then incubated at 25°C. After 10 to 15 days, fresh infective juveniles (IJs) and other developmental stages can be collected from the suspension (Figure 1C, D). Entomopathogenic nematodes (EPNs) can be effectively cultured through both in vitro and in vivo methods under laboratory conditions. In vivo methods require living, susceptible insect hosts from orders such as Coleoptera, Lepidoptera, and Diptera to support the maintenance, growth, and reproduction of EPNs. In the absence of suitable insect hosts, the in vitro approach can be employed for rearing EPNs. The in vitro method involves the use of various media, including Nutrient Bromothymol Blue Agar (NBTA), liver-kidney agar, and lipid agar media, to provide essential nutrients to the nematodes and their symbiotic bacteria [25].

3.1 Relationship between bacterium and nematode

Entomopathogenic nematodes (EPNs) and their symbiotic bacteria engage in a mutualistic relationship, with each benefiting from the other. The nematode depends on the bacterium to kill its insect host, create a favorable environment for its development by producing antibiotics that inhibit competing microorganisms, and break down host tissues into nutrients. In return, the bacterium relies on the nematode for protection from external environmental threats, for facilitating penetration into the host’s hemocoel, and for inhibiting the host’s antibacterial defenses. This mutualistic association exhibits a high level of species specificity between the nematode and its bacterial partner [14]. It is well-documented that EPNs are naturally present in a variety of agro-ecosystems. However, direct observation of insect cadavers infected with EPNs is challenging, and the patchy distribution of infective juveniles (IJs) in soil often prevents their detection using standard nematode extraction methods (Figure 2) [26].

Figure 2.

The life cycle of Photorhabdus and Xenorhabdus [26].

3.2 Multiplication of EPN

In vivo and in vitro methods are commonly employed for large-scale production of entomopathogenic nematodes (EPNs) [27]. However, in vitro mass production requires significant capital investment and specialized technical expertise. For a biological agent to be deemed successful, it is crucial that the host insect can be reared at a cost-effective rate and that the resulting product has a reasonably long shelf life. In contrast, although in vivo production of biocontrol agents necessitates lower financial investment and less technical expertise, scaling up the production process remains challenging, thereby making economic viability difficult to achieve [28]. Traditionally, in vivo mass rearing of EPNs has been conducted using the White trap method, initially developed by White in 1927 and subsequently modified [29]. Initially, the Baermann apparatus was used to recover infective juveniles (IJs) from cultures. Later, a novel method and apparatus were developed to harvest nematodes from a mixture of charcoal and fecal samples, based on the migratory behavior of parasitic nematodes in their third larval stage (Figure 3) [30].

Figure 3.

(A) The infective juvenile (IJ) stage of the nematode seeks out a suitable host for infection by penetrating the host’s hemolymph and releasing the pathogenic bacteria it carries. Within the insect host, the nematodes undergo development and reproduction, with multiple reproductive cycles occurring depending on the host’s size. As the host’s resources become depleted, a new generation of infective juveniles is produced, which then emerge and actively seek out new hosts to infect with the pathogenic bacteria. (B) Photographs depicting the emergence of entomopathogenic infective juveniles from Galleria mellonella (waxworm larvae) on the left and Acheta domestica (crickets) on the right [30].

3.3 Isolation of symbiotic bacteria

Symbiotic bacteria were isolated from infected cadavers. Infected larva is surface sterilized by dipping into 95% ethanol. Larva is cut open with sterile forceps, and the hemolymph is streaked on Macconkey agar plates. Primary phase colonies appeared in red, bright pink color, while secondary phase colonies ranged from light yellow to brown color.

3.4 Solid media

Foam pieces of about 1 cm cubic size are used to enhance the surface area and aeration in the system. The foam pieces are impregnated with the bacteriological medium (Wout’s medium) and packed in 500 ml Erlenmeyer flasks before autoclaving. The bacteria are inoculated onto the foam pieces and allowed to attain proper stage for nematode inoculation. Pure culture of appropriate EPN strain is inoculated on the foam pieces. Such systems were optimized for large-scale production using large autoclavable plastic bags, and the first commercial production of EPNs was based on this system.

3.5 Liquid media

Commercial-level EPN production has to employ liquid culture systems involving fermentation technology. In this system, provision of adequate levels of oxygen and removal of carbon dioxide and other waste gaseous material must be regulated so that it does not damage the delicate, relatively large, reproducing EPNs or interfere with copulation [14]. The in vitro liquid culture of entomopathogenic nematodes (EPNs) begins in Erlenmeyer shake flasks, which provide necessary aeration and agitation. Following the initial phase of liquid culture, the process is scaled up to 5–20 L desktop bioreactors and subsequently to industrial-scale bioreactors ranging from 80 to 1000 L (Figure 4) [31].

Figure 4.

Mass production of Entomopathogenic nematodes in different solid media: Multiplication of Heterorhabditis indica after 30 days of nematode inoculation on: (a) Modified dog biscuit medium; (b) modified Wout’s medium I; (c) modified Wout’s medium III; (d) Wout’s medium; and (e) multiplication of Heterorhabditis indica after 60 days of nematode inoculation on modified dog biscuit medium [31].

In the cultivation of monoxenic liquid cultures of the bacto-helminthic complex, the production medium, comprising essential nutrients, chicken tissue, and lipids, was adjusted to physiological pH and pre-inoculated with a 5% culture of Pseudomonas luminescens. This culture was a 24-hour growth maintained at 28°C and 150 rpm in 2× nutrient broth (NB). The inoculation process was conducted in Sartorius Stedim Biostat bioreactors, specifically A+ with capacities of 2 and 5 L, and B+ with a 10 L capacity, with respective working volumes of 1.5, 4, and 7 L. The bioreactors were operated under the following conditions: a temperature of 28°C, agitation at 200 rpm, a pH of 7.30, and dissolved oxygen at 30%. Concurrently, nematodes were surface sanitized using a 0.125% Hyamine solution and subsequently rinsed with sterile tap water to eliminate residual Hyamine. Upon reaching the early stationary phase of bacterial growth (20–24 hours), the bioreactors were inoculated with the sanitized nematodes at a density of approximately 2000 infective juveniles (IJs)/mL, and cultivation continued at 28°C with a pH of 7.3. During the initial phase of bacterial growth, agitation was maintained at 200 rpm across all bioreactors, after which it was reduced to 100 rpm in bioreactor A+ and to 150 and 200 rpm in bioreactor B+, respectively. Following nematode inoculation, dissolved oxygen levels were sustained at 30% during bacterial growth and increased to 40% for nematode mass production. The bioreactors were operated in batch mode for a duration of 7–8 days [32].

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4. Formulations

Nematodes intended for long-term storage and transportation to field sites can be formulated using various techniques. The formulation process should be carried out when the nematodes are fully active, either in motion or slightly desiccated, to reduce their mobility [33]. Since entomopathogenic nematodes (EPNs) are live, multicellular organisms, their quality tends to deteriorate over time, regardless of the formulation method, due to the depletion of their internal reserves. This decline is primarily attributed to the consumption of lipid reserves by the nematodes [34]. Two primary strategies are utilized to improve the suitability of formulations for the storage and transportation of nematodes. The first strategy involves the use of inert carriers, such as sponge or vermiculite, which ensure sufficient gaseous exchange. The second strategy is the induction of partial anhydrobiosis, a process that reduces the metabolic activity of the nematodes, thereby conserving their energy reserves [35].

4.1 Advantages of EPN

  • High host search ability.

  • Compatible with pesticides and other biocontrol agents of insect pests.

  • Capability of causing mortality of insects within 48–72 hours after infection.

  • Ability for mass multiplication on artificial media.

  • EPNs are viable for up to 3–6 months when refrigerated at 4 to 10°C.

  • EPNs can be easily applied in field using conventional sprayers and do not require any special equipment.

  • EPNs are safe for humans, vertebrates, and non-target organisms.

  • EPNs are exempt from registration requirements in many countries.

4.2 Disadvantages of EPN

  • Intolerance or mortality of the free-living stages of nematodes to low humidity and extreme temperatures.

  • Nematodes need to be protected from ultraviolet light, which soon reduces the infectivity and may kill the infective stages.

  • The relatively slow rate at which microorganisms kill their hosts, compared to chemical insecticides, has hindered their effectiveness and limited their acceptance among potential users.

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5. Efficacy of EPN against various insect pests

5.1 Efficacy of EPN against lepidopteran pests

The single-concentration screening assay revealed that, out of 10 entomopathogenic nematode (EPN) isolates tested, Heterorhabditis indica 1NBAIIH38, H. indica 2NBAIIH03, H. bacteriophora NBAIRH75, Steinernema carpocapsae NBAIRS59, and S. siamkayai 3 NBAIIS31 caused more than 50% mortality in 3rd instar larvae of Spodoptera frugiperda. Among these, H. indica 1NBAIIH38 exhibited the highest mortality (78%), while S. abbasi 2 NBAIIS49 caused the lowest mortality (20%). Further analysis indicated a significant variation in virulence among the EPN isolates, highlighting the importance of selecting the most virulent isolates for effective management of target insect pests [36].

Experiments conducted using the entomopathogenic nematode (EPN) Heterorhabditis indica (isolate: CICR Guava) on filter paper demonstrated its efficacy against Galleria mellonella and Spodoptera litura. The results indicated that H. indica caused 100% mortality in 5th instar larvae of G. mellonella at a treatment dose of 40 infective juveniles (IJs)/100 μl within 72 hours of infection. For S. litura, 100% mortality was recorded within 72 hours at a treatment dose of 100 IJs/100 μl in 3rd instar larvae, which exhibited higher susceptibility. These findings suggest a positive correlation between the concentration of nematode treatment, exposure time, and insect mortality in tobacco cutworm larvae. Thus, the H. indica isolate (CICR-Guava) can be recommended as a potential biocontrol agent for managing S. litura in the Vidarbha region [37]. The study also evaluated the susceptibility of S. litura to four locally isolated EPN species: H. indica, Heterorhabditis baujardi, Steinernema sangi, and Steinernema surkhetense. The LC50 values of H. indica at 72 hours post-incubation were 20.26 and 62.07 IJs/larva for the 3rd and 5th instars, respectively, and 913.34 IJs/pupa. The high mortality caused by H. indica and H. baujardi may be attributed to their high rate of penetration, typical of Heterorhabditids [38]. However, despite a lower penetration rate, S. sangi exhibited a similar level of pathogenicity to the Heterorhabditids. In laboratory bioassays comparing the biocontrol potential of Steinernema siamkayai (IIVR JNC01) with the commercially available H. indica (NBAIIH38 strain) against 3rd instar larvae of S. litura, Spilosoma obliqua, and Spoladea recurvalis, S. siamkayai (IIVR JNC01) caused 100, 100, and 85% mortality in these larvae, respectively [39].

The exploration and identification of native EPN species and strains adapted to local agro-climatic conditions is a crucial step in developing biocontrol agents for integrated pest management programs targeting major insect pests. The efficacy of eight native entomopathogenic nematodes (EPNs) against larvae of the hairy caterpillar (Euproctis sp.) and the shoot borer (Conogethes punctiferalis) was evaluated. Among the tested EPN isolates, all except Oscheius sp. IISR 08 caused 100% mortality in the larvae of the hairy caterpillar. Heterorhabditis sp. (IISR 01), Steinernema sp. (IISR 02), and Oscheius sp. (IISR 07 and 08) also caused 100% mortality in the larvae of the shoot borer [40].

5.2 Efficacy of EPN against hemipteran pests

The efficacy of native entomopathogenic nematodes (EPNs) against Planococcus citri was evaluated under laboratory conditions at various concentrations (80, 100, 150, 200 IJs/adult) and temperatures (20, 25, 30°C). The highest efficacy (68%) was achieved by Heterorhabditis indica 216-H at the highest concentration (200 IJs) at 25°C. Mortality rates were generally higher at 25°C compared to the other temperatures tested, and H. indica 216-H outperformed other EPN species at this temperature across all concentrations. These results suggest that H. indica 216-H holds significant potential for the control of P. citri [41]. In a separate study, the efficacy of Steinernema feltiae and H. bacteriophora against Dactylopius opuntiae was assessed. The results indicated that S. feltiae was more effective, causing a mortality rate of 98.8% in nymphs after 8 days of exposure, while H. bacteriophora resulted in a lower mortality rate of 83.8% [42].

5.3 Efficacy of EPN against coleopteran pests

S. siamkayai (IIVR JNC01 strain) caused 92.5% mortality of 2nd instar grubs of Myllocerus subfasciatus under laboratory conditions [39]. The bioefficacy of S. dharanaii (TFRIEPN-15) against white grub, Holotrichia rustica revealed that the minimum number of IJs, i.e., 300 Grub−1, causes 13.33% mortality in younger 6 days after the exposure, followed by 46.66% at the IJs population of 600 IJs Grub−1 [43]. The investigation on bioefficacy of entomopathogenic nematode, H. indica against white grub noted that mortality of white grub reached up to 73.34% at higher inoculums level (100 IJs/grub) after 120 hrs of application [44]. The study on the mortality of Capnodis tenebrionis larvae caused by different EPNs at different days after application exposed that after 1 D, larval mortality was 62.5, 52.5, 70.0, 50.0, and 0.00% with S. feltiae, S. carpocapsae, S. affine, H. bacteriophora, and control, respectively [45]. The efficacy of H. indica (DSM78) against naturally infested sugarcane white grub (H. serrata) was experimented under field conditions. The reduction in grub population due to EPN treatment ranged between 43.5 and 77% at field trial I and 30–78% at field trial II [46]. The bioefficacy of entomopathogenic nematodes against the white grub (Anomala communis) in potato was evaluated under field conditions. The highest larval mortality was observed 96 hours after treatment with Steinernema glaseri at a rate of 5 × 109 infective juveniles (IJs)/ha, followed by S. glaseri at 2.5 × 109 IJs/ha. The highest grub mortality (58.32%) was recorded with S. glaseri at 5 × 109 IJs/ha. Additionally, tuber damage was reduced to 24.99%, and the increase in yield was 15.78 t/ha when treated with S. glaseri at 5 × 109 IJs/ha [47].

5.4 Efficacy of EPN against dipteran pests

The experiments on the mortality of Bactrocera zonata and Bactrocera dorsalis adults when exposed to ten different species of entomopathogenic nematodes in Petri plates expressed that at all periods (12, 24, 48, and 72 h) and for both fly species, H. bacteriophora caused higher mortality than all other EPN species except S. carpocapsae [48].

5.5 Compatibility of EPN with insecticides

The compatibility of the entomopathogenic nematode Heterorhabditis indica with seven insecticides registered by the Central Insecticide Board was assessed under laboratory conditions. The results indicated that H. indica was most compatible with imidacloprid 17.8SL, as the highest percentage of live nematodes was observed 72 hours after exposure to this insecticide. H. indica also demonstrated compatibility with fipronil 5SC up to 48 hours of exposure. In contrast, less than 70% of H. indica remained viable after exposure to thiamethoxam 25WG, diafenthiuron 50WP, and cypermethrin 25EC, indicating that these insecticides were least compatible with the nematodes. Additionally, emamectin benzoate 5SG and chlorpyrifos 20EC were found to be incompatible with H. indica after 48 hours of exposure [49].

In laboratory bioassays, both S. carpocapsae and H. bacteriophora exhibited high tolerance to azadirachtin- and neonicotinoid-based insecticides, with survival rates ranging from 96.1 to 99.7 percent after 72 hours of exposure. S. carpocapsae demonstrated significantly higher virulence against last instar larvae of Agrotis ipsilon than H. bacteriophora at all tested concentrations. Larval mortality increased with both nematode concentration and exposure duration. At 160 infective juveniles (IJs) per larva, S. carpocapsae achieved cent percent mortality, while H. bacteriophora reached 90 per-cent after 72 hours. Combined applications of EPNs (80 IJs/larva) with either azadirachtin or neonicotinoid insecticides significantly enhanced larval mortality compared to individual treatments. Among combinations, EPNs with azadirachtin were more effective than those with neonicotinoids [50].

5.6 Compatibility of EPN with entomopathogenic fungi

The laboratory assays, Beauveria bassiana, Metarhizium anisopliae, and H. bacteriophora, showed significant pathogenicity against larval instars of Rhynchophorus ferrugineus. Synergistic effects were more frequent in H. bacteriophora + B. bassiana combinations, particularly in second and fourth instars at 0, 7, and 14 days, and in sixth instars at 0 and 7 days. Combined treatments reduced pupation, adult emergence, and egg hatching more than individual applications. Sub-lethal doses led to decreased larval weight, food consumption, frass production, and altered development. The H. bacteriophora + B. bassiana combination consistently showed the highest efficacy, especially against early instars [51].

Laboratory screening of EPNs for infectivity can be an important step in developing a biological control programme for a particular pest [52]. One of the main reasons for failure of EPNs as biological control of insect pests is the wrong choice of nematode species or strain [53] as results can vary greatly even among strains of the same species.

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6. Conclusions

Entomopathogenic nematodes (EPNs) have emerged as highly promising biological control agents for insect pest management, owing to their numerous attributes that align with the key characteristics of ideal biocontrol agents. These nematodes can be efficiently cultivated both in controlled laboratory conditions (in vitro) and within their natural environments (in vivo). Various types of insect pests that inhabit foliage and soil, such as coleopteran (white grub), dipteran (leaf miner), and lepidopteran (hairy caterpillar), can be effectively managed through the application of EPNs. What sets these nematodes apart is their compatibility with other crucial elements of Integrated Pest Management (IPM), including various chemical insecticides and other biological pesticides. This compatibility enhances their efficacy and versatility within integrated pest management strategies. The potential for developing and utilizing entomopathogenic nematodes as a sustainable option for pest control is substantial, highlighting their importance as a key component of environmentally friendly and holistic pest management.

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7. Future thrust

Future research and development should focus on advancing the practical applications of entomopathogenic nematodes (EPNs) in diverse agricultural systems. Efforts should aim to improve mass production techniques to make EPNs more cost-effective and accessible for large-scale use. Additionally, further studies on their compatibility with advanced Integrated Pest Management (IPM) tools, including novel formulations of biopesticides and precision agriculture technologies, will help maximize their effectiveness. Exploration of their potential in managing emerging pest threats and adapting to varying climatic conditions is also crucial. By leveraging innovative delivery systems and fostering collaborations between researchers, policymakers, and farmers, the full potential of EPNs as sustainable and environmentally sound pest control agents can be fully realized.

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Conflict of interest

The authors declare no conflict of interest.

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Written By

Thattankandy Meethal Aparna, Mukesh R. Siddhapara and Sonam Kumari

Submitted: 19 April 2025 Reviewed: 06 May 2025 Published: 25 July 2025

© 2025 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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