BIOTECHNOLOGICAL POTENTIAL OF ENTOMOPATHOGENIC MICROORGANISMS

GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 147 DOI: https://doi.org/10.5281/zenodo.17505117 BIOTECHNOLOGICAL POTENTIAL OF ENTOMOPATHOGENIC MICROORGANISMS Zukhra Y. Akhmedova1, Nigora Y. Zukhritdinova1, Mukhabbat Kh. Khashimova1, Sobir K. Sabirov1, Bobir A. Khamidov2, Firuza U. O’razova1 1 Institute of Zoology, Bogishamol st, 232b, Tashkent, 100053, Republic of Uzbekistan. 2 Karshi State University, Kuchabog st, 17, Karshi, 180100, Republic of Uzbekistan. Corresponding author: Zukhra Y. Akhmedova ([email protected]) ABSTRACT This review presents both widely used and promising subjects of entomology and microbiology-entomopathogenic microorganisms that possess unique physiological properties. The article summarises the achievements of scientific research concerning biological pest control and the development of biocontrol agents based on insecticidal formulations. Keywords: Bacteria, entomopathogenic microorganisms, micromycetes mycoinsecticides, viruses. Beneficial microorganisms are those that exert a positive effect on the environment and other organisms. These microorganisms play a pivotal role in the biological control of pests. Entomopathogenic microorganisms hold a particularly significant position, constituting the primary basis of biotechnology for producing entomopathogenic preparations used against harmful insects. For industrial-scale production of such biopreparations, bacteria, micromycetes, and viruses isolated from natural sources-as well as their metabolites-are primarily used. GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 148 Entomopathogenic bacteria are widely applied in agriculture. In addition to forming spores, these bacteria cause septicaemia in insects upon entry into their bodies and produce a range of endoand exotoxic compounds that enhance the effectiveness of biopreparations. Among bacterial pathogens affecting insect pests, several families have been identified: Bacillaceae, Pseudomonadaceae, Enterobacteriaceae, Streptococcaceae, and Micrococcaceae, some of which are virulent towards their respective hosts. Pathogens of arthropods such as Bacillus thuringiensis, B. sphaericus, B. cereus, and B. popilliae have been identified (Kalkha et al. 2014). The species Bacillus thuringiensis (Bt) has exhibited strong insecticidal properties in the control of insect pests and is known to act as an antagonist against phytopathogenic fungi due to its chitinolytic activity (Azizoglu 2019). Biopreparations have been developed on the basis of Bacillus thuringiensis, Lysinibacillus, Bacillus sphaericus, Paenibacillus spp, and Serratia entomophila for controlling dipteran and lepidopteran pests, including disease vectors (Lacey et al. 2015; Karimi et al. 2019). Rhizosphere bacteria isolated from soil, such as P. protegens and Streptomyces globisporus, were found to function as insect symbionts (Pronk et al. 2022). Entomopathogenic properties have also been confirmed in both bacteria and fungi such as Paraphaeosphaeria sporulosa, Setosphaeria rostrata GR1A, Aspergillus terreus, Fusarium fujikuroi, and Beauveria bassiana, making them effective agents against insect pests in agriculture (Watts et al. 2023). According to Salih Karabörklü et al. (2018), various organisms-viruses (acellular), bacteria (prokaryotic), fungi and protozoa (eukaryotic), and nematodes (multicellular)-have been evaluated for pest control, and the prospects for their use as entomopathogenic agents are discussed. Researchers such as Mayra Eleonora et al. (2025) have highlighted Bacillus thuringiensis as the most promising microorganism for targeting various insect orders using bacterial insecticides. Among the insecticidal compounds identified were homologous Cry toxins and non-homologous toxins, such as biosurfactants and macrocyclic lactones. Ricardo de Melo Katak et al. (2023) explored the considerable potential of microorganisms for controlling mosquito GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 149 populations and reducing their competency as vectors of diseases such as malaria, dengue fever, chikungunya, yellow fever, Zika virus, and filariasis. In their review, Li et al. (2022) presented a scientific basis for assessing the ecological biosafety of Bt transgenic plants, Bt microorganisms, and their expressed products. They also identified promising targets and methods for future Bt protein research and evaluation techniques. Scholars such as Mondal et al. (2023) investigated bacterial species associated with insect guts, the mechanisms of their symbiotic interactions, their role in insecticide resistance, and the practical applications of the gut microbiome in relation to host insects-thereby revealing mechanisms underpinning the evolution of pesticide resistance in insects. For the first time, Tang et al. (2022) provided comprehensive transcriptomic resources for N. pumilus. Their findings support the use of Novius species in biological control programmes and explore molecular mechanisms for adaptation in predation and mass rearing of N. pumilus and other Novius species, potentially contributing to pest management strategies. Researchers such as Loulou et al. (2023) isolated various bacterial species from rhabditid nematodes, including Acinetobacter sp, Alcaligenes sp, Bacillus cereus, Enterobacter sp, Kaistia sp, Lysinibacillus fusiformis, Morganella morganii subsp. morganii, Klebsiella quasipneumoniae subsp. quasipneumoniae, and Pseudomonas aeruginosa. All bacterial strains demonstrated high entomopathogenic activity, killing no less than 53.33% of Galleria mellonella larvae within 72 hours of exposure at a dose of 10⁶ CFU/larva. Entomopathogenic bacteria produce insecticidal proteins that function through mechanisms akin to those of proteolytic activation proteins. These proteins are integrated with Cry proteins in transgenic plants treated with Bacillus thuringiensis, contributing to enhanced insect resistance and expanded insecticidal spectrums (Chakroun et al. 2016). An insecticidal virulence factor was identified in entomopathogenic bacteria, with a particular focus on two insect-pathogenic genera: Photorhabdus (Proteobacteria: Enterobacteriaceae) and Bacillus (Firmicutes: Bacillaceae). It was noted that phage elements surrounding toxin genes distinguish GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 150 bacterial strains with virulence factors targeting insect neurons or neuromuscular junctions (Castagnola et al. 2014). Entomopathogenic micromycetes that cause diseases in insects possess valuable properties for the biological control of pests. These microorganisms are environmentally safe and harmless to humans and other ecosystem components. Fungi can be used as biopesticides effective against a broad spectrum of pests and serve as alternatives to chemical insecticides. The highest number of insecticidal compounds is produced by soil fungi, primarily from the genera Aspergillus, Penicillium, Trichoderma, Beauveria, and Metarhizium. According to Berestetskiy et al. (2021), various insecticidal compounds have been identified in promising fungal species. The efficiency of such research can be enhanced through high-throughput methods of fungal metabolite extraction and analysis via chromatography and mass spectrometry. Biopesticides have been developed from Lecanicillium lecanii and Beauveria bassiana to combat arthropod pests, and from Trichoderma harzianum to fight soilborne diseases (Hatting et al. 2019). Fungi such as B. bassiana, B. brongniartii, M. anisopliae, Lecanicillium lecanii, Hirsutella thompsonii, and Pochonia chlamydosporia infect arthropods and parasitic nematodes (Kiran et al. 2019). In the agrocenoses of the Turkestan region, two fungal species were isolated: B. bassiana (85%) and B. pseudobassiana (15%) (Rauza et al. 2024). The production of biologically active secondary metabolites by Penicillium species enhanced their entomopathogenic potential against pests (Nicoletti et al. 2023). Isolated entomopathogenic fungi such as Beauveria sp, Clonostachys sp, Cordyceps sp, Metarhizium sp, Purpureocillium sp, and Pochonia sp. were tested for virulence against Spodoptera litura (Liu et al. 2021). Using the baiting method, 51 entomopathogenic fungal strains were isolated-41 belonging to Metarhizium spp. and 10 to Beauveria spp, with baiting proving more effective than using nutrient media (Santos et al. 2022). Treatment with Beauveria bassiana A214b and Metarhizium anisopliae T331 caused 100% mortality of Galleria mellonella (L5) larvae within three GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 151 days, with LC50 of 3.33×10⁴ conidia/mL (Fofana et al. 2023). Isolates B. bassiana QB3.45, QB-3.46, and QB-3.428 caused mortality in immature Spodoptera frugiperda with egg mortality rates of 87.3%, 82.7%, and 79.3%, respectively, at a concentration of 1×10⁸ conidia/mL (Idrees et al. 2022). Spraying with Metarhizium anisopliae var. aridiorum at 0.25 and 0.5 L/ha caused mortality in 2nd–3rd instar Moroccan locusts at 42.3%, 82.3%, and 84.3% on day 7, and 61.3%, 95%, and 100% on day 21, respectively (Gapparov et al. 2013). Researchers synthesized silver nanoparticles using aqueous extracts of Beauveria bassiana, Metarhizium anisopliae, and Isaria fumosorosea, and studied their insecticidal effects against Plutella xylostella. The silver nanoparticles from Isaria fumosorosea showed the highest efficacy at a concentration of 0.691 mg/mL, with 78% mortality after 72 hours in second-instar larvae (Santos et al. 2023). Studies investigated the interaction between Metarhizium anisopliae, Beauveria bassiana, and the parasitoid Oomyzus sokolowskii on DBM larvae. Culture fluids at 10⁷ conidia/mL reduced parasitism in P. xylostella (Santos et al. 2006). Field experiments demonstrated that Metarhizium anisopliae and Beauveria bassiana isolates caused tick mortality and reduced reproduction (Fernandez et al. 2008). Laboratory studies (Fernandez et al. 2012; Min San et al. 2012) reviewed fungal-based biocontrol measures for ticks. A strain of Beauveria bassiana was used in wild rabbit burrows to control ticks, with a parasitic index (PI) reduction of 78.63% and 63.28% at 30 and 60 days post-treatment in spring; effectiveness dropped to 35.72% in summer (González et al. 2016). Pathogenicity of B. bassiana isolates against Rhipicephalus microplus females was confirmed, with B.bAT17 being highly virulent (LT50 and LT90 at 7–14 and 9–33 days, respectively, at 10⁹ conidia/mL) (Min San, Qiaoyun Ren et al. 2013). Metarhizium anisopliae, M. brunneum, and B. bassiana at 10⁶–10⁸ conidia/mL were pathogenic to D. albipictus larvae, with M. anisopliae and M. brunneum causing 74– 99% mortality, compared to 30–64% by B. bassiana (Sullivan et al. 2020). Larvae of different ages treated with M. brunneum F52 and B. bassiana GHA showed higher mortality with M. brunneum (Sullivan et al. 2022). Beauveria bassiana showed high GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 152 efficacy against apple sawflies in organic orchards, with 49.4–68.4% mycosis in treated larvae (Świergiel et al. 2016). Molecular studies using Beauveria and Metarhizium species revealed genes responsible for virulence, improving the effectiveness of mycoinsecticides in field pest control (Wang et al. 2017). For Anopheles larvae, researchers used Metarhizium anisopliae and B. bassiana spores in synthetic oil (ShellSol T), reducing pupation of An. gambiae by 39–50% (Bukhari et al. 2011). Sharma et al. (2020) isolated entomopathogenic bacteria (Serratia, Bacillus) and fungi (Beauveria, Clonostachys, Lecanicillium, Metarhizium, Purpureocillium) as effective biopesticides. Indian researchers (Kiran Kumar et al. 2019) developed bioformulations based on B. bassiana, B. brongniartii, M. anisopliae, L. lecanii, H. thompsonii, P. lilacinum, and P. chlamydosporia for use against arthropods and parasitic plants. From 240 samples from Australian vineyards, entomopathogenic fungi of three Beauveria species (B. bassiana, B. australis, B. pseudobassiana) and six Metarhizium species (M. guizhouense, M. robertsii, M. brunneum, M. flavoviride var, M. pingshaense, M. majus) were isolated (Korosi et al. 2019). From infected apple and cherry orchards (C. pomonella, C. cerasi), entomopathogenic fungi belonging to Aspergillus, Fusarium, and Alternaria spp. were identified (Akhmedova et al. 2024; Zukhritdinova et al. 2025). Within the subfamily Erynioideae of Entomophthoraceae, six genera (Erynia, Furia, Orthomyces, Pandora, Strongwellsea, Zoophthora) were found to affect mostly Diptera and Hemiptera, although few species were cultivated in vitro (Gryganskyi et al. 2024). Gonzalez et al. (2016) reviewed the use of fungal, bacterial, and viral entomopathogens in European greenhouses. Deka et al. (2021) noted that fungal, bacterial, viral, and nematode pathogens offer advantages over chemical pesticidesprecision, safety, and sustainability. Hyphomycete fungi are the most widely used entomopathogens in IPM systems for controlling B. tabaci, especially at stage 2 of infection (Ortiz-Urquiza et al. 2013). These fungi penetrate the insect cuticle and secrete detoxifying enzymes like lipases, esterases, catalases, cytochrome P450s, proteases, and chitinases (Ortiz-Urquiza and Keyhani, 2013). GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 153 Entomopathogenic viruses. Viral insecticides are biological preparations based on entomopathogenic viruses that selectively affect insect pests. They are safe for humans, warm-blooded animals, and beneficial entomofauna, making them an important tool in organic and integrated agriculture. The development of modern viral metagenomics has contributed to the discovery and study of viruses in various ecosystems. This approach provides new opportunities to explore interactions between insects and plants, which in turn may help optimize their management. In 36 hymenopteran parasitoids, two viruses were identified - Spodoptera exigua nucleopolyhedrovirus and Spodoptera frugiperda nucleopolyhedrovirus - which were used to treat pests such as Trichogramma pretiosum and Encarsia formosa. Fungi of the species Beauveria bassiana, Metarhizium anisopliae, Lecanicillium muscarium, and the bacterium Bacillus thuringiensis were also used (Koller et al. 2023). Viruses were detected in 54.8% of samples, containing dsRNA elements with viral characteristics. One RNA sequence corresponded to a genome of 5.2 kb from the Totiviridae family, designated as B. bassiana RNA virus BbRV1 (Herrero et al. 2012). New ISV-type viruses belonging to the Rhabdoviridae family were discovered in three specimens of the insect Riptortus pedestris. This study supports the development of potential biological control agents against insect pests (Guo et al. 2022). The effectiveness of the corn earworm virus used for biological control in maize cultivation areas against Xylella fastidiosa was studied, as well as its potential as a biopesticide (Picciotti et al. 2023). From larval samples of Hypera postica (alfalfa weevils), five virus species were isolated, along with four from Medicago sativa (alfalfa). A trophic relationship was identified between plant virus diversity and phytophagous pests (François et al. 2021). Strains of the nuclear polyhedrosis virus were isolated from dead larvae of Helicoverpa armigera (cotton bollworm) and Cydia pomonella (codling moth). The strains were active against pests, with a concentration of (1–2)×10¹² polyhedra per larva and showed high biological efficacy – up to 99.8% by the seventh GOLDEN BRAIN ISSN: 2181-4120 VOLUME 3 | ISSUE 16 | 2025 Multidisciplinary Scientific Journal October, 2025 154 day against larvae of both H. armigera and C. pomonella (Akhmedova et al. 2021, 2024). The effect of infection by the mycovirus Beauveria bassiana chrysovirus 2 (BbCV2) on host biological characteristics was studied. Infection with BbCV2 increased the growth rate of B. bassiana, spore production, and biomass, and also enhanced the ability of host fungi and their metabolites to inhibit phytopathogenic fungi (Li et al. 2024). According to Kotta-Loizou and Coutts (2017), B. bassiana isolates containing elements of Polymycoviridae mycoviruses were selected. Viral infection prevalence reached up to 54.8%. One RNA element matched a 5.2 kb genome of a previously undescribed member of the Totiviridae family, named B. bassiana RNA virus 1 (BbRV1). Acknowledgment This research work was carried out within the framework of the research program of the Institute of Zoology of the Academy of Sciences of the Republic of Uzbekistan for 2025-2029 “1.2. Creation of a digital information system of the animal world of the Bukhara and Navoi regions”, financed from the state budget. 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