Phenotypic and biochemical characterisation and pathogenicity assessment on Galleria mellonella L. (Lepidoptera: Pyralidae) of symbionts of the entomopathogenic nematode Heterorhabditis amazonensis Andalo et al., 2006
DOI:
https://doi.org/10.5433/1679-0359.2023v44n3p1047Keywords:
Microbial control, Photorhabdus, Symbiotic association, Virulence.Abstract
The objective of this study was to describe phenotypically and biochemically the symbiotic bacteria associated with three populations of Heterorhabditis amazonensis Andalo et al., 2006 (isolates: UEL-n 01, UEL-n 07, and UEL-n 08) and evaluate their pathogenicity on Galleria mellonella L. (Lepidoptera: Pyralidae) larvae. Bacteria were isolated by maceration of infective juveniles (IJs) and grown in culture medium (NBTA and MacConkey). The characterization of the bacteria was evaluated by employing motility test and biochemical tests like Gram staining, lipase activity, protease, and lecithinase. The production of antibiotics and bioluminescence was also evaluated. The pathogenicity was evaluated on the last instar larvae of G. mellonella at a concentration of 104 cells/mL. The bacteria from the three entomopathogenic nematodes isolates were positive for all biochemical tests except for lecithinase, and have presented bioluminescence when subjected to ultraviolet light, indicating that they belong to the genus Photorhabdus sp. Both were pathogenic to G. mellonella larvae causing 93.3 to 100.0% mortality.
Metrics
References
Adams, B. J., Fodor, A., Klein, M. G., Smith, H. L., Stackebrandt, E., & Stock, S. P. (2006). Biodiversity and systematics of nematode-bacterium entomopathogens. Biological Control, 37(1), 32-49. doi: 10.1016/S1049-9644(06)00126-5 DOI: https://doi.org/10.1016/j.biocontrol.2005.11.008
Almenara, D. P., Rossi, C., Neves, C. M. R., & Winter, C. E. (2012). Nematoides entomopatogênicos. In M. A. C. Silva Neto da, C. Winter, & C. Termignoni (Eds.), Tópicos avançados em entomologia molecular (pp. 1-40). São Paulo. DOI: https://doi.org/10.7594/revbio.06b.01
Andalo, V., Nguyen, K. B. & Moino, A. Jr. (2006). Heterorhabditis amazonensis n. sp. (Rhabditida: Heterorhabditidae) from Amazonas, Brazil. Nematology, 8(6), 853-867. doi: org/10.1163/156854106779799286 DOI: https://doi.org/10.1163/156854106779799286
Akhurst, R. (1980). Morphological and functional dimorphism in Xenorhabdus spp., bacteria symbiotically associated with the insect pathogenic nematodes Neoaplectana and Heterorhabditis. Journal of Genetic Microbiology, 121(2), 303-309. doi: 10.1099/00221287-121-2-303 DOI: https://doi.org/10.1099/00221287-121-2-303
Bird, A. F., & Akhurst, R. J. (1983). The nature of the intestinal vesicle in nematodes of the family Steinernematidae. International Journal of Parasitology, 13(6), 599-606. doi: 10.1016/S0020-7519(83)80032-0 DOI: https://doi.org/10.1016/S0020-7519(83)80032-0
Boemare, N. E., Akhurst R. J., & Mourant, R. G. (1993). Deoxyribonucleic acid relatedness between Xenorhabdus spp. (Enterobacteriaceae), symbiotic bacteria of entomopathogenic nematodes, with a proposal to transfer Xenorhabdus luminescens to a new genus, Photorhabdus. International Journal of Systematic Bacteriology, 43(2), 249-255. doi: 10.1099/00207713-43-2-249 DOI: https://doi.org/10.1099/00207713-43-2-249
Boemare, N. E., Laumond, C., & Mauléon, H. (1996). The nematode-bacterium complexes: biology, life cycle and vertebrate safety. Biocontrol Science and Technology, 6(1), 333-345. doi: 10.1080/09583159631316 DOI: https://doi.org/10.1080/09583159631316
Boemare, N., & Akhurst, R. (1988). Biochemical and physiological characterization of colony form variants in Xenorhabdus spp. (Enterobactereaceae). International Journal of Systematic Bacteriology, 134(3), 751-761. doi: 10.1099/00221287-134-3-751 DOI: https://doi.org/10.1099/00221287-134-3-751
Bowen, D., & Ensign, J. (1998). Purification and characterization of a highmolecular-weight insecticidal protein complex produced by the entomopathogenic bacterium Photorhabdus luminescens. Applied and Environmental Microbiology, 64(8), 3029-3035. doi: 10.1128/AEM.64.8.3029-3035.1998 DOI: https://doi.org/10.1128/AEM.64.8.3029-3035.1998
Bowen, D., Blackburn, M., Rocheleau, T., Grutzmacher, C., & Ffrench-Constant, R. H. (2000). Secreted proteases from Photorhabdus luminescens: separation of the extracellular proteases from the insecticidal Tc toxin complexes. Insect Biochemistry and Molecular Biology, 30(2), 69-74. doi: 10.1016/s0965-1748(99)00098-3 DOI: https://doi.org/10.1016/S0965-1748(99)00098-3
Brenner, D. J. (1999). Additional genera of enterobacteriaceae. In M. Dworkin, Falkow, S., Rosenberg, E., Schleifer, K.-H., & Stackebrand, E. (Eds.), The prokaryotes: an evolving electronic resource for the microbiological community (3nd ed., release 3.4). New York.
Burman, M. (1982). Neoaplectana carpocapse: toxin production by axenic insect parasitic nematodes. Nematológica, 78(1), 62-70. doi: 10.1163/187529282X00510 DOI: https://doi.org/10.1163/187529282X00510
Costa, J. R. V., Rossi, J. R., Marucci, S. C., Alves, E. C. C., Volpe, H. X. L., Ferraudo, A. S., Lemos, M. V. F., & Desidério, J. A. (2010). Atividade tóxica de isolados de Bacillus thuringiensis a larvas de Aedes aegypti (L.) (Diptera: Culicidae). Neotropical Entomology, 39(5), 757-766. doi: 10.1590/S1519-566X2010000500015 DOI: https://doi.org/10.1590/S1519-566X2010000500015
Dowds, B. C. A., & Peters, A. (2002). Virulence mechanisms. In R. Gaugler (Ed.), Entomopathogenic nematology (pp. 79-98). New York. DOI: https://doi.org/10.1079/9780851995670.0079
Ferreira, D. F. (2011). Sisvar: a computer statistical analysis system. Ciência e Agrotecnologia, 35(6), 1039-1042. doi: 10.1590/S1413-70542011000600001 DOI: https://doi.org/10.1590/S1413-70542011000600001
Ferreira, T., Van Reenen, C., Pagès, S., Tailliez, P., Malan, A. P., & Dicks, L. M. T. (2013). Photorhabdus luminescens subsp. noenieputensis subsp. nov., a symbiotic bacterium associated with a novel Heterorhabditis species related to Heterorhabditis indica. International Journal of Systematic and Evolutionary Microbiology, 63(5), 1853-1858. doi: 10.1099/ijs.0.044388-0 DOI: https://doi.org/10.1099/ijs.0.044388-0
Forst, S., Dowds, B., Boemare, N., & Stackebrandt, E. (1997). Xenorhabdus and Photorhabdus spp.: bugs that kill bugs. Annual Review of Microbiology, 51(1), 47-72. doi: 10.1146/annurev.micro.51.1.47 DOI: https://doi.org/10.1146/annurev.micro.51.1.47
Fukruksa, C., Yimthin, T., Suwannaro, J. 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), 1-10. doi: 10.1186/s13071-017-2383-2 DOI: https://doi.org/10.1186/s13071-017-2383-2
Guerra, B. E., Chacón, J. G., Muñoz, J. E., & Caicedo, M. A. (2014). Evaluación de la patogenicidad de Xenorhabdus spp. nativos en Colombia. Revista Colombiana de Biotecnologia, 16(1), 111-118. doi: 10.15446/rev.colomb.biote.v16n1.44277 DOI: https://doi.org/10.15446/rev.colomb.biote.v16n1.44277
Guide, B. A., Andaló, V., Ferreira, D. G., Alves, V. S., Fernandes, T. A. P., & Neves, P. M. O. J. (2022). First report and biological characteristics of Heterorhabditis amazonensis in the state of Paraná, Brazil. Brazilian Journal of Biology, 84(1), e262374. doi: 10.1590/1519-6984.262374 DOI: https://doi.org/10.1590/1519-6984.262374
Han, R. C., & Ehlers, R. U. (2000). Pathogenicity, development, and reproduction of Heterorhabditis bacteriophora and Steinernema carpocapsae under axenic in vivo conditions. Journal Invertebrate Pathology, 75(1), 55-58. doi: 10.1006/jipa.1999.4900 DOI: https://doi.org/10.1006/jipa.1999.4900
Jarosz, J., Balcerzak, M., & Skrzypek, H. (1991). Involvement of larvicidal toxin in pathogenesis of insect parasitism with the rhabditoid nematodes Steinernema feltiae and Heterorhabditis bacteriophora. Entomophaga, 36(1), 361-368. doi: 10.1007/BF02377940 DOI: https://doi.org/10.1007/BF02377940
Koppenhöfer, H. S. (2007). Bacterial symbionts of Steinernema and Heterorhabditis. Nematology Monographs and Perspectives, 2(1), 735-808. doi: 10.1163/ej.9789004152939.i-816.43 DOI: https://doi.org/10.1163/ej.9789004152939.i-816.43
Kushwah, J., Kumar, P., Garg, V., & Somvanshi, V. S. (2016). Discovery of a highly virulent strain of Photorhabdus luminescens ssp. akhurstii from Meghalaya, India. Indian Journal of Microbiology, 57(1), 125-128. doi: 10.1007/s12088-016-0628-y DOI: https://doi.org/10.1007/s12088-016-0628-y
Liu, W., Ye, W., Wang, Z., Wang, X., Tian, S., Cao, H., & Lian, J. (2006). Photorhabdus luminescens toxin-induced permeability change in Manduca sexta and Tenebrio molitor midgut brush border membrane and in unilamellar phospholipid vesicle. Environmental Microbiology, 8(5), 858-870. doi: 10.1111/j.1462-2920.2005.00972.x DOI: https://doi.org/10.1111/j.1462-2920.2005.00972.x
Molina, J. P. A., & López, N. J. C. (2001). Producción in vivo de três entomonematodos con dos sistemas de infección en dos hospedantes. Revista Colombiana de Entomologia, 27(1-2), 73-78. DOI: https://doi.org/10.25100/socolen.v27i1.9670
Orozco, R. A., Hill, T., & Stock, P. S. (2013). Characterization and phylogenetic relationships of Photorhabdus luminescens subsp. sonorensis (γ-Proteobacteria: Enterobacteriaceae), the bacterial symbiont of the entomopathogenic nematode Heterorhabditis sonorensis (Nematoda: Heterorhabditidae). Current Microbiology, 66(1), 30-39. doi: 10.1007/s00284-012-0220-6 DOI: https://doi.org/10.1007/s00284-012-0220-6
Paracer, S., & Ahmadjian, V. (2000). Bacterial associations of bactéria, protozoa and animals. In S. Paracer, & V. Ahmadjian (Eds.), Symbiosis: an introduction to biological associations (pp. 33-50). New York.
Schmidt, T. M., Bleakley, B., & Nealson, K. H. (1988). Characterization of an extracellular protease from the insect pathogen Xenorhabdus luminescens. Apply of Environmental Microbiology, 54(11), 2793-2797. doi: 10.1128/aem.54.11.2793-2797.1988 DOI: https://doi.org/10.1128/aem.54.11.2793-2797.1988
Sierra, G. (1957). A simple method for the detection of lipolytic activity of micro-organisms and some observations on the influence of the contact between cells and fatty substrates. Antonie van Leeuwenhoek, 23(1), 15-22. doi: 10.1007/BF02545855 DOI: https://doi.org/10.1007/BF02545855
Singh, S., Eric, M., & Floyd, I. (2012). Characterization of Photorhabdus luminescens growth for the rearing of the beneficial nematode Heterorhabditis bacteriophora. Indian Journal of Microbiology, 52(3), 325-331. doi: 10.1007/s12088-011-0238-7 DOI: https://doi.org/10.1007/s12088-011-0238-7
Stehr, F., Kretschmar, M., Kröger, C., Hube, B., & Schäfer, W. (2003). Microbial lipases as virulence factors. Journal of Molecular Catalysis B: Enzymatic, 22(5-6), 347-355. doi: 10.1016/S1381-1177(03)00049-3 DOI: https://doi.org/10.1016/S1381-1177(03)00049-3
Thaler, J. O., Baghdiguian, S., & Boemare, N. E. (1995). Puri® cation and characterization of xenorhabdicin, a phage tail-like bacteriocin, from the lysogenic strain F1 of Xenorhabdus nematophilus. Applied and Environ- mental Microbiology, 61(5), 2049-2052. doi: 10.1128/aem.61.5.2049-2052.1995 DOI: https://doi.org/10.1128/aem.61.5.2049-2052.1995
Whaley, D. N., Dowell, V. R., Wanderlinder, L. M., & Lombard, G. L. (1982). Gelatin agar medium for detecting gelatinase production by anaerobic bacteria. Journal of Clinical Microbiology, 16(2), 224-229. doi: 10.1128/jcm.16.2.224-229.1982 DOI: https://doi.org/10.1128/jcm.16.2.224-229.1982
White, G. F. (1927). A method for obtaining infective nematodes larvae from cultures. Science, 66(1709), 302-303. doi: 10.1126/science.66.1709.302-a DOI: https://doi.org/10.1126/science.66.1709.302.b
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Semina: Ciências Agrárias
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
O Copyright dos manuscritos publicados pertence ao periódico. Como são publicados em um periódico de acesso aberto, eles estão disponíveis gratuitamente, para uso privado ou para fins educacionais e não comerciais.
A revista tem o direito de fazer, no documento original, alterações referentes às normas lingüísticas, ortografia e gramática, com o objetivo de garantir as normas padrão do idioma e a credibilidade da revista. No entanto, respeitará o estilo de escrita dos autores.
Quando necessário, alterações conceituais, correções ou sugestões serão encaminhadas aos autores. Nesses casos, o manuscrito deve ser submetido a uma nova avaliação após revisão.
A responsabilidade pelas opiniões expressas nos manuscritos é inteiramente dos autores.
