Bioactivity of endosymbiotic bacterial supernatants from local entomopathogenic nematodes against Galleria mellonella L. (Lepidoptera: Pyralidae) larvae

Abstract

Entomopathogenic nematodes (EPNs) and their symbiotic bacteria stand out as environmentally friendly alternatives due to their high pathogenicity against harmful insects. This study aimed to evaluate the insecticidal effects of cell-free supernatants (CFSs) derived from the symbiotic bacteria of local EPN isolates on larvae of Galleria mellonella (Lepidoptera: Pyralidae) under controlled conditions. The bacterial associates of EPNs were isolated from newly harvested infective juveniles (IJs) of EPNs and cell-free supernatants (CFSs) belonging to Xenorhabdus bovienii (MCB-8) and Photorhabdus luminescens subsp. kayaii (AVB-15) strains were applied to 4th instar larvae by spraying. Larval mortalities were recorded at 24, 48, 72, and 96 hours post-treatment. Both bacterial supernatants were able to cause mortality on the larvae. The mortality rates varied depending on the exposure times and increased sharply after 96 hours of exposure. The lowest mortality (2.5%) was obtained at 24 hours post-treatment, while mortalities reached 80.0% (X. bovienii) and 86.2% (P. luminescens) after 96 hours of exposure to CFSs, respectively. The findings revealed that the culture filtrates of symbiotic bacteria from local EPN isolates have strong insecticidal activity against G. mellonella larvae.

Keywords

• Entomopathogenic nematodes
• Symbiotic bacteria
• Sustainability

Yerel entomopatojen nematodlardan elde edilen endosimbiyotik bakteriyel süpernatantların Galleria mellonella L. (Lepidoptera: Pyralidae) larvalarına karşı biyoaktivitesi

Abstract

Entomopatojen nematodlar (EPN) ve simbiyotik bakterileri, zararlı böceklere karşı yüksek patojeniteleri sayesinde çevre dostu alternatifler arasında öne çıkmaktadır. Bu çalışma, yerel EPN izolatlarından elde edilen simbiyotik bakterilere ait hücresiz süpernatantların (HS) Galleria mellonella (Lepidoptera: Pyralidae) larvaları üzerindeki insektisidal etkilerini kontrollü koşullar altında değerlendirmeyi amaçlamıştır. EPN'lerin bakteriyel simbiyontları, EPN'lerin yeni hasat edilmiş infektiv juvenillerinden (IJ'ler) izole edilmiş ve Xenorhabdus bovienii (MCB-8) ve Photorhabdus luminescens subsp. kayaii (AVB-15) suşlarına ait hücresiz süpernatantlar (HS'ler) 4. evre larvalara püskürtme yoluyla uygulanmıştır. Larva ölümleri uygulamadan 24, 48, 72 ve 96 saat sonra kaydedilmiştir. Her iki bakteriyel süpernatant da larvalarda ölüme neden olabilmiştir. Ölüm oranları maruz kalma sürelerine bağlı olarak değişiklik göstermiş ve uygulamadan 96 saat sonra keskin bir şekilde artmıştır. En düşük ölüm oranı (%2,5) uygulama sonrası 24 saat sonra elde edilirken, uygulamadan 96 saat sonra ölüm oranları sırasıyla %80,0 (X. bovienii) ve %86,2 (P. luminescens)'ye ulaşmıştır. Bulgular, yerel EPN izolatlarından elde edilen simbiyotik bakterilerin hücre içermeyen süpernatantlarının G. mellonella larvalarına karşı güçlü bir insektisidal etkiye sahip olduğunu ortaya koymuştur.

Keywords

• Entomopatojen nematodlar
• Simbiyotik bakteriler
• Sürdürülebilirlik

References

1. Abd-Elgawad, M. M., Askary, T. H., & Coupland, J. (2017). Biocontrol agents: entomopathogenic and slug parasitic nematodes. CAB International, Boston. pp 629.
2. Adithya, S., Shivaprakash, M., & Sowmya, E. (2020). Evaluation of insecticidal activity of entomopathogenic bacteria Photorhabdus and Xenorhabdus against shoot and fruit borer Earias vittella (Lepidoptera: Noctuidae) of vegetable crops. Journal of Entomology and Zoology Studies, 8, 2343-2348. https://doi.org/10.22271/phyto.2020.v9.i3x.11516
3. Boemare, N. E., & Akhurst, R. J. (1988). Biochemical and physiological characterization of colony-forming variants in Xenorhabdus spp.(Enterobacteriaceae). Microbiology, 134(3), 751-761. https://doi.org/10.1099/00221287-134-3-751
4. Cimen, H., Touray, M., Gulsen, S. H., & Hazir, S. (2022). Natural products from Photorhabdus and Xenorhabdus: mechanisms and impacts. Applied Microbiology and Biotechnology, 106(12), 4387-4399. https://doi.org/10.1007/s00253-022-12023-9
5. Da Silva, W. J., Pilz-Júnior, H. L., Heermann, R., & da Silva, O. S. (2020). The great potential of entomopathogenic bacteria Xenorhabdus and Photorhabdus for mosquito control: a review. Parasites & vectors, 13(1), 376. https://doi.org/10.1186/s13071-020-04236-6
6. Dunphy, G. B., & Webster, J. M. (1988). Lipopolysaccharides of Xenorhabdus nematophilus (Enterobacteriaceae) and their haemocyte toxicity in non-immune Galleria mellonella (Insecta: Lepidoptera) larvae. Microbiology, 134(4), 1017-1028. https://doi.org/10.1099/00221287-134-4-1017
7. Erdem, E., Yüksel, E., & Canhilal, R. (2024). Evaluation of cell-free supernatants from the symbiotic bacteria of entomopathogenic nematodes for controlling the Colorado potato beetle [Leptinotarsa decemlineata (Say)(Coleoptera: Chrysomelidae)]. Journal of Plant Diseases and Protection, 131(3), 731-742. https://doi.org/10.1007/s41348-024-00894-1
8. Eroglu, C., Cimen, H., Ulug, D., Karagoz, M., Hazir, S., & Cakmak, I. (2019). Acaricidal effect of cell-free supernatants from Xenorhabdus and Photorhabdus bacteria against Tetranychus urticae (Acari: Tetranychidae). Journal of Invertebrate Pathology, 160, 61-66. https://doi.org/10.1016/j.jip.2018.12.004

9. ffrench-Constant*, R. H., & Bowen, D. J. (2000). Novel insecticidal toxins from nematode-symbiotic bacteria. Cellular and Molecular Life Sciences CMLS, 57(5), 828-833. https://doi.org/10.1007/s000180050044
10. Fukruksa, C., Yimthin, T., Suwannaroj, M., Muangpat, P., Tandhavanant, S., Thanwisai, A., & Vitta, A. (2017). Isolation and identification of Xenorhabdus and Photorhabdus bacteria associated with entomopathogenic nematodes and their larvicidal activity against Aedes aegypti. Parasites & Vectors, 10(1), 440. https://doi.org/10.1186/s13071-017-2383-2
11. Goodrich‐Blair, H., & Clarke, D. J. (2007). Mutualism and pathogenesis in Xenorhabdus and Photorhabdus: two roads to the same destination. Molecular microbiology, 64(2), 260-268. https://doi.org/10.1111/j.1365-2958.2007.05671.x
12. Hasan MA, Ahmed S, Mollah MM, Lee D, Kim Y (2019) Variation in pathogenicity of different strains of Xenorhabdus nematophila; Differential immunosuppressive activities and secondary metabolite production. Journal of Invertebrate Pathology, 166, 107221–107231. https://doi.org/10.1016/j.jip.2019.107221
13. Hazir, S., Shapiro-Ilan, D. I., Bock, C. H., Hazir, C., Leite, L. G., & Hotchkiss, M. W. (2016). Relative potency of culture supernatants of Xenorhabdus and Photorhabdus spp. on growth of some fungal phytopathogens. European Journal of Plant Pathology, 146(2), 369-381. https://doi.org/10.1007/s10658-016-0923-9
14. Kushwah, J., & Somvanshi, V. S. (2015). Photorhabdus: a microbial factory of insect-killing toxins. VC Kalia (Ed). Microbial Factories: Biodiversity, Biopolymers, Bioactive Molecules (2nd edition, pp. 235-240). New Delhi Springer. https://doi.org/10.1007/978-81-322-2595-9_15
15. Metwally, H. M., Hafez, G. A., Hussein, M. A., Hussein, M. A., Salem, H. A., & Saleh, M. M. E. (2012). Low cost artificial diet for rearing the greater wax moth, Galleria mellonella L.(Lepidoptera: Pyralidae) as a host for entomopathogenic nematodes. Egyptian Journal of Biological Pest Control, 22(1), 15.
16. Nermuť, J., Hristov, Y., Podrábská, V., Kraml, A., Skoková Habuštová, O., Konopická, J., Doležal, P., & Půža, V. (2025). Potential of cell-free supernatants of Xenorhabdus and Photorhabdus bacteria for the control of virus-transmitting aphids (Myzus persicae). Journal of Applied Microbiology, 136(6), lxaf150. https://doi.org/10.1093/jambio/lxaf150
17. Ogier, J. C., Akhurst, R., Boemare, N., & Gaudriault, S. (2023). The endosymbiont and the second bacterial circle of entomopathogenic nematodes. Trends in Microbiology, 31(6), 629-643. https://doi.org/10.1016/j.tim.2023.01.004
18. Ozakman, Y., & Eleftherianos, I. (2020). Immune interactions between Drosophila and the pathogen Xenorhabdus. Microbiological Research, 240, 126568. https://doi.org/10.1016/j.micres.2020.126568
19. Ruiu, L., Virdis, B., Mura, M. E., Floris, I., Satta, A., & Tarasco, E. (2017). Oral insecticidal activity of new bacterial isolates against insects in two orders. Biocontrol Science and Technology, 27(7), 886-902. https://doi.org/10.1080/09583157.2017.1355964
20. Shi, H., Zeng, H., Yang, X., Liu, Z., & Qiu, D. (2013). An insecticidal protein from Xenorhabdus ehlersii stimulates the innate immune response in Galleria mellonella. World Journal of Microbiology and Biotechnology, 29(9), 1705-1711. https://doi.org/10.1007/s11274-013-1333-1
21. Shi, H., Zeng, H., Yang, X., Zhao, J., Chen, M., & Qiu, D. (2012). An insecticidal protein from Xenorhabdus ehlersii triggers prophenoloxidase activation and hemocyte decrease in Galleria mellonella. Current microbiology, 64(6), 604-610. https://doi.org/10.1007/s00284-012-0114-7
22. Singh, A. K., Kumar, M., Ahuja, A., Vinay, B. K., Kommu, K. K., Thakur, S., & Parihar, M. (2022). Entomopathogenic nematodes: a sustainable option for insect pest management. A Rakshit, VS Meena, PC Abhilash, BK Sarma, HB Singh, L Fraceto, M Parihar and K Singh (Eds.), Biopesticides (pp. 73-92). Woodhead. https://doi.org/10.1016/B978-0-12-823355-9.00007-9
23. Tarasco, E., Fanelli, E., Salvemini, C., El-Khoury, Y., Troccoli, A., Vovlas, A., & De Luca, F. (2023). Entomopathogenic nematodes and their symbiotic bacteria: from genes to field uses. Frontiers in Insect Science, 3, 1195254. https://doi.org/10.3389/finsc.2023.1195254
24. Ulug, D., Touray, M., Gulsen, S. H., Cimen, H., Hazir, C., Bode, H. B., & Hazir, S. (2024). A taste of a toxin paradise: Xenorhabdus and Photorhabdus bacterial secondary metabolites against Aedes aegypti larvae and eggs. Journal of Invertebrate Pathology, 205, 108126. https://doi.org/10.1016/j.jip.2024.108126
25. Vicente-Díez, I., Blanco-Pérez, R., González-Trujillo, M. D. M., Pou, A., & Campos-Herrera, R. (2021). Insecticidal effect of entomopathogenic nematodes and the cell-free supernatant from their symbiotic bacteria against Philaenus spumarius (Hemiptera: Aphrophoridae) nymphs. Insects, 12(5), 448. https://doi.org/10.3390/insects12050448
26. Vitta, A., Thimpoo, P., Meesil, W., Yimthin, T., Fukruksa, C., Polseela, R., Bandid, M., Sarunporn, T., & Thanwisai, A. (2018). Larvicidal activity of Xenorhabdus and Photorhabdus bacteria against Aedes aegypti and Aedes albopictus. Asian Pacific Journal of Tropical Biomedicine, 8(1), 31-36. https://doi.org/10.4103/2221-1691.221134
27. Yang, J., Zeng, H. M., Lin, H. F., Yang, X. F., Liu, Z., Guo, L. H., Yuan, J., & Qiu, D. W. (2012). An insecticidal protein from Xenorhabdus budapestensis that results in prophenoloxidase activation in the wax moth, Galleria mellonella. Journal of Invertebrate Pathology, 110(1), 60-67. https://doi.org/10.1016/j.jip.2012.02.006
28. Yuksel, E., & Canhilal, R. (2019). Isolation, identification, and pathogenicity of entomopathogenic nematodes occurring in Cappadocia Region, Central Turkey. Egyptian Journal of Biological Pest Control, 29(1), 40. https://doi.org/10.1186/s41938-019-0141-9
29. Yüksel, E. (2022). Biocontrol potential of endosymbiotic bacteria of entomopathogenic nematodes against the tomato leaf miner, Tuta absoluta (Meyrick)(Lepidoptera: Gelechiidae). Egyptian Journal of Biological Pest Control, 32(1), 135. https://doi.org/10.1186/s41938-022-00633-4
30. Yüksel, E., Imren, M., Özdemir, E., Bozbuğa, R., & Canhilal, R. (2022). Insecticidal effect of entomopathogenic nematodes and the cell-free supernatants from their symbiotic bacteria against different larval instars of Agrotis segetum (Denis & Schiffermüller)(Lepidoptera: Noctuidae). Egyptian Journal of Biological Pest Control, 32(1), 54. https://doi.org/10.1186/s41938-022-00555-1
31. Zobar, D. & Kıvan, M. (2022). Bazı entomopatojen fungus türlerinin Capnodis tenebrionis Linnaeus, 1758 (Coleoptera: Buprestidae)’in belirli dönemleri üzerinde patojeniteleri. Mustafa Kemal Üniversitesi Tarım Bilimleri Dergisi, 27(3), 469-476. https://doi.org/10.37908/mkutbd.1071813