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University of São Paulo “Luiz de Queiroz” College of Agriculture Taxonomic studies of Macrochelidae mites (Acari: Mesostigmata) and their potential use to control Stomoxys calcitrans and Musca domestica (Diptera: Muscidae) Letícia Henrique de Azevedo Thesis presented to obtain the degree of Doctor in Science. Area: Entomology Piracicaba 2017 Letícia Henrique de Azevedo Agronomist Taxonomic studies of Macrochelidae mites (Acari: Mesostigmata) and their potential use to control Stomoxys calcitrans and Musca domestica (Diptera: Muscidae) Advisor: Prof. Dr. GILBERTO JOSÉ DE MORAES Thesis presented to obtain the degree of Doctor in Science. Area: Entomology Piracicaba 2017 2 This work is especially dedicated To GOD My mother Maria Paula Henrique da Silva Azevedo My husband Diego Luis Prado My brother Matheus Henrique de Azevedo For all the love and support offered to me To Prof. Dr. Gilberto José de Moraes For his valuable guidance, friendship and recognition of my work Special thanks 3 ACKNOWLEDGEMENTS I am grateful to God for supporting me in my way. To Escola Superior de Agricultura “Luiz de Queiroz” (ESALQ), Universidade de São Paulo (USP), for giving me the opportunity to conduct my studies. I am especially grateful to Dr. Raphael de C. Castilho (FCAV/UNESP, Brazil), for his great friendship, valuable suggestions and advices, and for all his support during my Doctorate studies. To Professor Dr. Carlos H. W. Flechtmann (ESALQ) for all your lessons, all his assistance provided whenever needed and help with translations of German, Latin and French publications. To Dr. Rowan M. Emberson, retired professor of Entomology at the University of Lincoln (Canterbury, New Zealand), for his great collaboration in chapter 1 and 2 of this thesis, by sending me the file with the listing of species of world Macrochelidae, in other words, for turning possible the conduction of those studies. To Dr. Gerald W. Krantz for important personal information provided and for the examination specimens of the new species described in this thesis. To “Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Gado de Corte”, Campo Grande, State of Mato Grosso do Sul (Brazil) for providing the facilities for the conduction of the predation tests with the stable fly and horn fly. To Dr. Paulo H. D. Cançado and Dr. Antonio Thadeu M. Barros, for the guidance and valuable suggestions for the success of the tests conducted at EMBRAPA. To Taciany F. S. Dominghetti and her husband André Dominghetti for the help and the hospitality during the time spent in Campo Grande. To EMBRAPA´s laboratory technicians Ronaldo Souza and Marco Antonio Silva, for the help in the laboratory activities. To Dr. Bruce Halliday for his great help in the interpretation of the spermathecal apparatus of new species, for promptly providing me with several hard-to-get publications I needed and for the stimulating contacts. To Diana M. Rueda for kindly assisting me with the statistical analysis; To Springer International Publishing for granting permission to reproduce the title “Macrochelid Mites (Mesostigmata: Macrochelidae) as Biological Control Agents” as an introductory chapter of this thesis; 4 To “Programa de Pós-Graduação em Entomologia”, especially to its coordinator, Dr. Fernando L. Cônsoli, and the secretary Andrea V. Sintoni, for supporting my activities. To USP COMUT-Librarians for providing copies of many papers. To Lásaro V. F. da Silva (ESALQ/USP, Brazil), for logistic support and laboratory help. To my friends of Acarology Laboratory of ESALQ/USP, Camila Tavares, Diana M. Rueda, Fernanda de C. N. Esteca, Geovanny S. P. Barroso, Grazielle F. Moreira, Jandir C. Santos, João B. da Cruz, João Paulo Z. Narita, João Pedro Martin, Marcela M. R. da Silva, Márcia Daniela, Marina F. C. Barbosa, Michele Ennes, Renan V. da Silva, Reham Abo- Shnaf, Tiago Casaroto. Special thanks to Marielle de M. Berto and Murilo P. Ferreira for the outstanding collaboration in mite samplings and laboratory tests. To the laboratory and administrative staff members of “Departamento de Entomologia e Acarologia” of ESALQ/USP, José L. F. Piedade, Josenilton L. Mandro, Maria Marta Coletti, Rosângela A. da Silva. To Eriana F. Prado and Wilson F. Prado, my parents in law, who always supported me and encouraged my studies. To Débora P. Zamboni, my sister in law, and her husband Giuliano Zamboni for all the affection. To my great friend Marielle de M. Berto for all the affection and friendship. To all my family and especially to the relatives of my husband, for their warm support and love. To “Conselho Nacional de Desenvolvimento Científico e Tecnológico” (CNPq, Brazil) for the scholarship that supported me during my Doctoral studies. And to those who in one way or another helped me in this work. Thank you! 5 RESUMO Estudos taxonômicos de ácaros Macrochelidae (Acari: Mesostigmata) e o potencial de uso destes para o controle de Stomoxys calcitrans e Musca domestica (Diptera: Muscidae) Macrochelidae Vitzthum é um dos mais abundantes e diversos grupos de ácaros predadores que encontrados em excrementos ou em carcaças de animais em decomposição. Algumas espécies dessa família tem mostrado potencial como agentes de controle biológico. Stomoxys calcitrans L. tem sido um problema sério enfrentado por pecuaristas em alguns estados brasileiros e tem causado perdas significativas no setor pecuarista. Já Musca domestica L. pode ser vetora de várias patógenos de seres humanos e de animais. Os objetivos do estudo relatado neste documento foram a elaboração de um catálogo taxonômico de espécies de Macrochelidae do mundo, determinar as espécies de macroquelídeos encontradas em microhabitats em que a mosca-dos-estábulos é geralmente encontrada nas fazendas do Estado de São Paulo, e avaliar o efeito de diferentes espécies de macroquelídeos como possíveis agentes de controle de moscas nocivas. Um total de 520 espécies de 23 gêneros são relatadas no catálogo. No total, 1359 espécimes foram coletados de esterco de gado e serapilheira em diferentes regiões do Estado de São Paulo. Estes representaram dez espécies de três gêneros e uma nova espécie, que foi descrita. Nos testes em laboratório, Macrocheles n. sp. apresentou maiores taxas de predação e oviposição alimentando-se de larvas de S. calcitrans (23,8 larvas predadas e oviposição de cerca de 4 ovos por dia, respectivamente). Para Macrocheles n. sp., o desenvolvimento da fase imatura (ovo-adulto) foi concluído em 1,3 ± 0,01, 1,3 ± 0,01 e 1,5 ± 0,03 dias alimentando-se de ovos de S. calcitrans, M. domestica e de Protorhabditis sp., respectivamente. A fecundidade foi maior quando alimentado com Protorhabditis sp. (77,2 ± 5,1 ovos / fêmea) do que com S. calcitrans (58,0 ± 5,9) e M. domestica (55,0 ± 5,3). Os resultados deste estudo sugerem que Macrocheles n. sp. é um promissor agente de controle biológico de S. calcitrans e M. domestica e que poderia ser criado de forma massal com o uso de Protorhabditis sp. ou ovos de M. domestica. Palavras-chave: Controle biológico; Catálogo; Ácaros predadores; Mosca-dos- estábulos 6 ABSTRACT Taxonomic studies of Macrochelidae mites (Acari: Mesostigmata) and their potential use to control Stomoxys calcitrans and Musca domestica (Diptera: Muscidae) The family Macrochelidae Vitzthum is one of the most abundant and diverse groups of predatory mites living in excrements or decomposing animals. Some species of this family have shown good potential as biological control agents. Stomoxys calcitrans L. is a serious problem that has been faced by cattle raisers in some Brazilian states, causing significant losses. Musca domestica L. may vector several pathogens that affect humans and animals. The aims of the study reported in this document were the preparation of a taxonomic catalog of the world macrochelid species, to determine the macrochelid species in microhabitats where the stable fly is usually found in commercial cow raising farms in São Paulo State, and to evaluate of the effect of different macrochelid species as control agents of harmful muscid flies. A total of 520 species of 23 genera are reported in the catalog. In total, 1359 specimens were collected from cow manure and litter in different regions of São Paulo State. These represented ten species of three genera and one new species, which was described. In laboratory tests, Macrocheles n. sp. presented higher predation and oviposition rates on larvae of S. calcitrans (23.8 larvae consumed and laid about 4 eggs per day, respectively). For Macrocheles n. sp., total immature development (egg- adult) was completed in 1.3 ± 0.01, 1.3 ± 0.01 and 1.5 ± 0.03 days on eggs of S. calcitrans and M. domestica and on Protorhabditis sp., respectively. Fecundity was higher on Protorhabditis sp. (77.2 ± 5.1 eggs/female) than on S. calcitrans (58.0 ± 5.9) and M. domestica (55.0 ± 5.3). Results of this study suggest that Macrocheles n. sp. is a promising biological control agent of S. calcitrans and M. domestica, and that it could be mass reared with the use of Protorhabditis sp. or eggs of M. domestica. Keywords: Biological control; Catalog; Predatory mite; Stable fly 7 1. INTRODUCTION Macrochelid mites (Mesostigmata: Macrochelidae) as biological control agents (With permission of Springer International Publishing) Biological control of plant pests using predatory mites has been extensively practiced in many countries, especially in protected crops. The predators most often used for that purpose belong to the Phytoseiidae (Gerson et al. 2003; Beaulieu and Weeks 2007), especially because these are usually by far the most common plant inhabiting group of predatory mites. The family Macrochelidae Vitzthum (Figure 1), of the order Mesostigmata, is one of the most abundant and diverse groups of predatory mites living in excrements or decomposing animals (Vitzthum 1930; Koehler 1999; Behan-Pelletier 2003; Gerson et al. 2003; Braig and Perotti 2009; Lindquist et al. 2009; Perotti and Braig 2009). Filipponi (1964) divided the macrochelids into three main groups, fimicolous, humicolous and insecticolous, referring to their association with excrements, humus or insects (mainly as carriers of these mites).