Avaliação de dietas para criação de Galleria mellonella l. (Lepidoptera: Pyralidae) e multiplicação de nematoides entomopatogênicos
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Data
2022-11-23
Autores
Souza, Jennifer Kawane Aparecida
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Universidade Estadual Paulista (Unesp)
Resumo
Galleria mellonella é um inseto modelo aceito em estudos de interações patógenohospedeiro e sobre compostos antimicrobianos. É oferecida como hospedeiro em diversos laboratórios no mundo, por sua facilidade de manuseio, aceitação de técnicas de infecção e pelo seu sistema imune inato, semelhante ao de mamíferos. Essa espécie é empregada em pesquisas relacionadas a saúde humana, animal e no desenvolvimento de técnicas e tecnologias de controle biológico de pragas. Em nematologia agrícola, as lagartas de G. mellonella são oferecidas aos nematoides entomopatogênicos (NEPs), como hospedeiro padrão. Embora a criação da espécie de mariposa seja considerada fácil, a nutrição desses insetos é um fator limitante, pois quando inadequada influencia diretamente nos processos de infecção, virulência e multiplicação de Heterorhabditis e Steinernema (gêneros das famílias de NEPs). Em compor uma dieta que beneficie o ciclo de vida de G. mellonella, estimulando a fase larval, aumentando o tamanho das lagartas e o número de adultos na população final, com uma prole saudável e bem desenvolvida, essa pesquisa tem por objetivo, estudar a resposta de indivíduos de G. mellonella quando submetidos a uma nova dieta artificial e verificar se as lagartas alimentadas por ela, podem comprometer, favorecer, ou não serem significantes, nos processos de infecção, virulência e multiplicação de criações de nematoides entomopatogênicos em laboratório. Para isso duas etapas foram utilizadas nesta pesquisa, a primeira corresponde as avaliações biométricas, demográficas e dos parâmetros de vida e fertilidade para as lagartas de G. mellonella nutridas com diferentes dietas artificiais, sendo o tratamento 1: favos secos umedecidos com solução de mel a 75%, tratamento 2: dieta composta de leite em pó integral, mel, farinha de aveia, gérmen de trigo e proteína de soja e tratamento 3: dieta padrão criada por Guerra (1973). As populações foram avaliadas por 8 gerações. Para a segunda etapa, foram verificadas as influências da alimentação das lagartas sobre os processos de infecção, virulência e multiplicação de nematoides entomopatogênicos, em que os tratamentos correspondem as lagartas da oitava geração de G. mellonella padronizadas em comprimento e largura para cada dieta em 6° instar, oferecidas a cinco espécies de nematoides entomopatogênicos, sendo elas: Heterorhabditis bacteriophora, H. amazonensis, Steinernema puertoricense, S. rarum e S. feltiae. Sobre as dietas, pode-se verificar que as lagartas alimentadas pela dieta adaptada de Jorjão et al. (2018), baseada em Guerra (1973) e modificada de Parra (1998) apresentaram os melhores índices de desenvolvimento e médias positivas para grande parte dos parâmetros avaliados, enquanto a dieta correspondente ao tratamento 1 demonstrou médias negativas sobre o desenvolvimento da espécie G. mellonella. A dieta do tratamento 3 se mostrou nesta pesquisa uma boa opção de alimentação a lagartas da espécie, porém seu uso não é recomendado para criações com finalidade de hospedeiras a nematoides entomopatogênicos, visto que as lagartas alimentadas por essa dieta, apresentam baixa produção de juvenis infectantes. Para as avaliações da influência da alimentação das lagartas sobre os processos realizados pelos NEPs, a dieta correspondente ao tratamento 2 se apresentou favorável a infecção, virulência e multiplicação para às cinco espécies avaliadas.
Galleria mellonella is a model insect accepted in studies of pathogen-host interactions and on antimicrobial compounds. It is offered as a host in several laboratories around the world, for its ease of handling, acceptance of infection techniques and for its innate immune system, similar to that of mammals. This species is used in research related to human and animal health and in the development of techniques and technologies for biological pest control. In agricultural nematology, caterpillars of G. mellonella are offered to entomopathogenic nematodes (NEPs) as a standard host. The NEPs belong to the Heterorhabditidae and Steinernematidae families, which are able to control insect pests of annual and perennial crops in a period of 24 to 48 hours after spraying them on crops. Although the creation of the moth species is considered easy, the nutrition of these insects is a limiting factor, because when inadequate it directly influences the processes of infection, virulence and multiplication of Heterorhabditis and Steinernema (genera of the NEP families). In composing a diet that benefits the life cycle of G. mellonella, stimulating the larval stage, increasing the size of caterpillars and the number of adults in the final population, with a healthy and well-developed offspring, this research aims to study the response of individuals of G. mellonella when submitted to a new artificial diet and to verify if the caterpillars fed by it, can compromise, favor, or not be significant, in the processes of infection, virulence and multiplication of creations of entomopathogenic nematodes in the laboratory. For this, two stages were used in this research, the first corresponds to biometric, demographic and life and fertility parameters assessments for G. mellonella caterpillars fed with different artificial diets, being treatment 1: dry combs moistened with honey solution at 75%, treatment 2: diet composed of whole milk powder, honey, oat flour, wheat germ and soy protein and treatment 3: standard diet created by Guerra (1973). Populations were evaluated for 8 generations. For the second stage, the influences of the caterpillars' feeding on the processes of infection, virulence and multiplication of entomopathogenic nematodes were verified, in which the treatments correspond to the caterpillars of the eighth generation of G. mellonella standardized in length and width for each diet in 6 th instar, offered to five species of entomopathogenic nematodes, namely: Heterorhabditis bacteriophora, H. amazonensis, Steinernema puertoricense, S. rarum and S. feltiae. Regarding the diets, it can be seen that the caterpillars fed by the adapted diet showed the best development rates and positive averages for most of the parameters evaluated, while the diet corresponding to diet 1 showed negative averages on the development of the species G. mellonella. Diet 3 was shown in this research to be a good option for feeding caterpillars of the species, but its use is not recommended for creations with the purpose of hosting entomopathogenic nematodes, since caterpillars fed by this diet have low production of infective juveniles. For the evaluations of the influence of the caterpillars' feeding on the processes carried out by the NEPs, the diet corresponding to treatment 2 was favorable to infection, virulence and multiplication for the five species evaluated.
Galleria mellonella is a model insect accepted in studies of pathogen-host interactions and on antimicrobial compounds. It is offered as a host in several laboratories around the world, for its ease of handling, acceptance of infection techniques and for its innate immune system, similar to that of mammals. This species is used in research related to human and animal health and in the development of techniques and technologies for biological pest control. In agricultural nematology, caterpillars of G. mellonella are offered to entomopathogenic nematodes (NEPs) as a standard host. The NEPs belong to the Heterorhabditidae and Steinernematidae families, which are able to control insect pests of annual and perennial crops in a period of 24 to 48 hours after spraying them on crops. Although the creation of the moth species is considered easy, the nutrition of these insects is a limiting factor, because when inadequate it directly influences the processes of infection, virulence and multiplication of Heterorhabditis and Steinernema (genera of the NEP families). In composing a diet that benefits the life cycle of G. mellonella, stimulating the larval stage, increasing the size of caterpillars and the number of adults in the final population, with a healthy and well-developed offspring, this research aims to study the response of individuals of G. mellonella when submitted to a new artificial diet and to verify if the caterpillars fed by it, can compromise, favor, or not be significant, in the processes of infection, virulence and multiplication of creations of entomopathogenic nematodes in the laboratory. For this, two stages were used in this research, the first corresponds to biometric, demographic and life and fertility parameters assessments for G. mellonella caterpillars fed with different artificial diets, being treatment 1: dry combs moistened with honey solution at 75%, treatment 2: diet composed of whole milk powder, honey, oat flour, wheat germ and soy protein and treatment 3: standard diet created by Guerra (1973). Populations were evaluated for 8 generations. For the second stage, the influences of the caterpillars' feeding on the processes of infection, virulence and multiplication of entomopathogenic nematodes were verified, in which the treatments correspond to the caterpillars of the eighth generation of G. mellonella standardized in length and width for each diet in 6 th instar, offered to five species of entomopathogenic nematodes, namely: Heterorhabditis bacteriophora, H. amazonensis, Steinernema puertoricense, S. rarum and S. feltiae. Regarding the diets, it can be seen that the caterpillars fed by the adapted diet showed the best development rates and positive averages for most of the parameters evaluated, while the diet corresponding to diet 1 showed negative averages on the development of the species G. mellonella. Diet 3 was shown in this research to be a good option for feeding caterpillars of the species, but its use is not recommended for creations with the purpose of hosting entomopathogenic nematodes, since caterpillars fed by this diet have low production of infective juveniles. For the evaluations of the influence of the caterpillars' feeding on the processes carried out by the NEPs, the diet corresponding to treatment 2 was favorable to infection, virulence and multiplication for the five species evaluated.
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Palavras-chave
criação em laboratório, Heterorhabditis, técnica in vivo, NEPs, Steinernema, Creation in laboratory, Technique in vivo