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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
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Efeito de plantas hospedeiras no desenvolvimento e reprodu??o do parasitoide Palmistichus elaeisis (Hymenoptera: Eulophidae) / Effect of host plants on the development and reproduction of the parasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae)

Camilo, Silma da Silva 21 February 2014 (has links)
Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-23T11:46:15Z No. of bitstreams: 2 silma_silva_camilo.pdf: 711117 bytes, checksum: b6de81038261d9e969a9528685c405e8 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-23T11:48:16Z (GMT) No. of bitstreams: 2 silma_silva_camilo.pdf: 711117 bytes, checksum: b6de81038261d9e969a9528685c405e8 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-23T11:48:44Z (GMT) No. of bitstreams: 2 silma_silva_camilo.pdf: 711117 bytes, checksum: b6de81038261d9e969a9528685c405e8 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) / Made available in DSpace on 2014-12-23T11:48:44Z (GMT). No. of bitstreams: 2 silma_silva_camilo.pdf: 711117 bytes, checksum: b6de81038261d9e969a9528685c405e8 (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) Previous issue date: 2014 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / Objetivou-se avaliar a influ?ncia da alimenta??o do hospedeiro e dos adultos do parasitoide Palmistichus elaeisis Delvare & LaSalle, 1993 (Hymenoptera: Eulophidae) em seus par?metros biol?gicos e capacidade de parasitismo. No primeiro experimento, analisou-se a efici?ncia do parasitismo e a biologia da prole deste parasitoide em pupas de Thyrinteina arnobia Stoll 1782 (Lepidoptera: Geometridae) criadas em plantas de Psidium guajava L. ou Eucalyptus cloeziana F. Muell (Myrtaceae). Ovos de T. arnobia foram colocados em sacos de tecido tipo organza envolvendo galhos de plantas de P. guajava e E. cloeziana, at? as lagartas atingirem a fase de pupa. Trinta pupas de cada tratamento foram individualizadas em tubos de vidro e expostas ao parasitismo por quatro f?meas de P. elaeisis. Foi avaliada a emerg?ncia da prog?nie por pupa; a porcentagem de parasitismo, pupas mortas e de adultos de T. arnobia emergidos; a dura??o do ciclo de vida (ovo-adulto), a longevidade, a raz?o sexual, o tamanho da c?psula cef?lica e do corpo do parasitoide. A porcentagem de parasitismo, a emerg?ncia de P. elaeisis por pupa, a longevidade das f?meas, o tamanho da c?psula cef?lica e do corpo dos machos do parasitoide foram menores quando seu hospedeiro foi criado em plantas de eucalipto. No segundo experimento objetivou-se avaliar o efeito da alimenta??o com p?len e n?ctar de flores de Eucalyptus sp. e de plantas daninhas encontradas em cultivos deste, no desenvolvimento e reprodu??o de P. elaeisis. Parasitoides adultos machos e f?meas foram individualizados em tubos de vidro para a avalia??o da longevidade. Receberam os tratamentos: flores das aster?ceas Bidens pilosa L., Emilia fosbergii Nicolson, Galinsoga parviflora Cav., Sonchus oleraceus L. e Taraxacum officinale F.H. Wigg., de Eucalyptus sp., uma got?cula de mel e sem alimenta??o. Em outra etapa, seis f?meas foram transferidas para potes pl?sticos juntamente com uma pupa de Tenebrio molitor Linnaeus 1758 (Coleoptera: Tenebrionidae) para ser parasitada, sendo empregados os mesmos tratamentos anteriores. As avalia??es foram: sobreviv?ncia de f?meas, porcentagem de parasitismo e emerg?ncia, dura??o do ciclo de vida, n?mero de indiv?duos emergidos por pupa, raz?o sexual, tamanho da c?psula cef?lica e do corpo da prole. O uso de flores de plantas daninhas e de Eucalyptus sp. como fonte de alimento permitiu que os insetos se reproduzissem. A alimenta??o de P. elaeisis com mel proporcionou maior longevidade e sucesso reprodutivo. A priva??o de alimento reduziu a longevidade do parasitoide e comprometeu sua reprodu??o. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Produ??o Vegetal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2014. / ABSTRACT The objective was evaluate the influence of feeding host and adults parasitoid Palmistichus elaeisis Delvare & LaSalle, 1993 (Hymenoptera: Eulophidae) on their biological parameters and parasitism capacity. In the first experiment were analyzed the efficiency of parasitism and biology of this parasitoid in pupae of Thyrinteina arnobia 1782 (Lepidoptera: Geometridae) when reared on plants of Psidium guajava L. or Eucalyptus cloeziana F. Muell (Myrtaceae). Eggs of T. arnobia were placed organza bags involving branches of plants of P. guajava and E. cloeziana until the larvae reach the pupal stage. Thirty pupae from each treatment were placed in glass tubes and exposed to parasitism by four females of P. elaeisis. It was evaluated emergence of the parasitoid progeny per pupa, the percentage of parasitism, dead pupae and adults of T. arnobia emerged; duration of the life cycle (egg to adult), longevity, sex ratio, the size of the head capsule and body of the parasitoid. The percentage parasitism, emergence of P. elaeisis per pupa, the female longevity, the size of the head capsule and body of the parasitoid males were smaller when its host was created in eucalyptus plants. In the second experiment, the objective was to evaluate the effect of feeding with pollen and nectar from Eucalyptus sp. flowers and weeds found in these cultivations, in the development and reproduction of P. elaeisis. Parasitic adult males and females were individualized in glass tubes for evaluation of this longevity. They received the treatments: flowers of Asteraceae Bidens pilosa L., Emilia fosbergii Nicolson, Galinsoga parviflora Cav., Sonchus oleraceus L. e Taraxacum officinale F.H. Wigg., Eucalyptus sp., a droplet of honey and without feeding. In another stage, six females were transferred to plastic pots with a pupa of Tenebrio molitor Linnaeus 1758 (Coleoptera: Tenebrionidae) to be parasitized, they were used previous treatments. The evaluations were: survival of females, parasitism and emergence, duration of the life cycle, number of adults per pupa, sex ratio, size of the head capsule and body of the offspring. The use of flowers weeds and Eucalyptus sp. as a food source has allowed the reproduction of the insects. The feeding of P. eleaisis with honey showed provided greater longevity and reproductive success. The food deprivation reduced the longevity of the parasitoid and hindered this reproduction.
2

Testes gen?ticos e biol?gicos de baculov?rus coletados em Helicoverpa armigera (H?BNER) (Lepidoptera: Noctuidae) no Brasil / Genetic and biological tests of baculovirus collected in Helicoverpa armigera (H?bner) (Lepidoptera: Noctuidae) in Brazil

Costa, Victor Hugo Duarte da January 2015 (has links)
Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-05T14:00:08Z No. of bitstreams: 2 victor_hugo_duarte_costa.pdf: 749112 bytes, checksum: 673a3fb7e223fa45ab64a0cf8798bedf (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-05T18:02:07Z (GMT) No. of bitstreams: 2 victor_hugo_duarte_costa.pdf: 749112 bytes, checksum: 673a3fb7e223fa45ab64a0cf8798bedf (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Made available in DSpace on 2016-01-05T18:02:07Z (GMT). No. of bitstreams: 2 victor_hugo_duarte_costa.pdf: 749112 bytes, checksum: 673a3fb7e223fa45ab64a0cf8798bedf (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Previous issue date: 2015 / Funda??o de Amparo ? Pesquisa do estado de Minas Gerais (FAPEMIG) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / Lagartas da esp?cie Helicoverpa armigera foram identificadas no Brasil atacando extensas ?reas de produ??o agr?cola. Algumas caracter?sticas fazem deste inseto uma das principais pragas a sistemas agr?colas do mundo, o que torna este ataque um perigo eminente ?s ?reas de produ??o do pa?s. Estudos com os insetos coletados nestas ?reas s?o essenciais para que seja desenvolvido um manejo adequado. Estudos de laborat?rio podem selecionar poss?veis pr?ticas de manejo a fim de combater surtos de pragas no campo. Uma das formas de manejar o ambiente de forma segura ? adotar o Manejo Integrado de Pragas, onde se destaca o controle biol?gico utilizando baculov?rus. A lagarta de H. armigera ? suscept?vel a estirpes de baculov?rus, por?m ? necess?rio verificar esta suscetibilidade. Sendo assim, os objetivos deste trabalho foram selecionar estirpes aut?ctones de baculov?rus t?xicas as lagartas desta esp?cie. Foram realizados testes gen?ticos e biol?gicos entre as estirpes aut?ctones, comparando-as com o produto comercial Gemstar?, de origem norte americana. A an?lise comparativa do sequenciamento gen?tico realizada para os genes LEF-8 e LEF-9 revelaram que os isolados locais est?o, estreitamente, relacionados com esp?cies de baculov?rus da Austr?lia e da ?ndia. Todos os isolados testados possibilitaram o controle de lagartas de terceiro instar de H. armigera. Por?m, analises biol?gicas da concentra??o letal m?dia (CL50) e do tempo letal m?dio (TL50) variaram entre os isolados testados. O isolado HearNPV-BR2 apresentou os melhores resultados de CL50 e TL50. Ademais, este ? o primeiro registro da ocorr?ncia da esp?cie HearNPV-BR2 no Brasil, e suas propriedades inseticidas assinalam que a mesma pode ser ?til para a fabrica??o de bioinseticidas para o controle de H. armigera no pa?s. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Produ??o Vegetal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015. / ABSTRACT Larvea of Helicoverpa armigera species were identified in Brazil attacking extensive agricultural production areas. Some features make this one of the main insect pests in agricultural systems in the world, which makes this attack an imminent danger to agricultural production in the country. Studies with insects collected in these areas are essential for developed proper management. Laboratory studies can select possible management practices to combat pest outbreaks in the field. One way to manage the secure environment is to adopt the Integrated Pest Management, highlighting the biological control using baculovirus. The H. armigera larvea is susceptible to strains of baculovirus but it is necessary check this susceptibility. Therefore the objectives of this work were to select indigenous strains of toxic baculovirus larvea of this species. Genetic and biological tests were carried out between indigenous strains, comparing them with the commercial product Gemstar?, origin from North American (USA). Comparative analysis of genetic sequencing performed for the LEF-8 and LEF-9 genes, revealed that indigenous strains are closely related to species of baculovirus of the Australia and India. All strains tested allowed control of the third instar of the H. armigera larvea. However, biological analysis of median lethal concentration (LC 50) and median lethal time (LT50) varied among the strains tested. The HearNPV-BR2 strain showed the best results of CL50 and TL50. Moreover, this is the first record of the occurrence of HearNPV-BR2 species in Brazil and its insecticidal properties indicate that BR2 may be useful for the manufacture of biological insecticides for H. armigera control in the country.
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Defici?ncias nutricionais de macronutrientes em mudas de eucalipto afetam o consumo alimentar, desenvolvimento e fitofagia de Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae)? / Nutritional deficiencies of macronutrients in eucalyptus seedlings affect food consumption, development, and phytophagy of Podisus nigrispinus (Dallas, 1851) (Hemiptera: Pentatomidae)?

Vieira, Estela Rosana Dur?es 26 February 2015 (has links)
Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-05T12:51:59Z No. of bitstreams: 2 estela_rosana_duraes_vieira.pdf: 510975 bytes, checksum: a22fd6b1844ea89c0f2c4098c9736ff6 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-05T17:51:18Z (GMT) No. of bitstreams: 2 estela_rosana_duraes_vieira.pdf: 510975 bytes, checksum: a22fd6b1844ea89c0f2c4098c9736ff6 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Made available in DSpace on 2016-01-05T17:51:18Z (GMT). No. of bitstreams: 2 estela_rosana_duraes_vieira.pdf: 510975 bytes, checksum: a22fd6b1844ea89c0f2c4098c9736ff6 (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Previous issue date: 2015 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / A busca por esclarecimentos direcionados ? fitofagia de percevejos predadores pode fornecer subs?dios que beneficiam os programas de controle biol?gico, tornando-os mais eficazes. Sabendo-se que os macronutrientes est?o envolvidos em diversos processos fisiol?gicos dos insetos, o objetivo deste trabalho foi identificar o efeito da omiss?o de macronutrientes em mudas de eucalipto no desenvolvimento e consumo alimentar do Podisus nigrispinus. Al?m disso, procurou-se certificar se a fitofagia, provoca perdas nas mudas. Este estudo foi conduzido em casa de vegeta??o e os tratamentos consistiram na omiss?o individual de nitrog?nio, f?sforo, pot?ssio, c?lcio, magn?sio e enxofre na solu??o nutritiva, onde as mudas de eucalipto estavam sendo conduzidas. Pupas de Tenebrio molitor L. (Coleoptera: Tenebrionidae), foram oferecidas ad libitum aos percevejos predadores. Comprovou-se a import?ncia da complementa??o nutricional em plantas para o desenvolvimento e preda??o do P. nigrispinus. A alimenta??o em plantas deficientes em nitrog?nio e magn?sio diminuiu a efici?ncia de preda??o deste zoofit?fago. E a omiss?o de c?lcio, magn?sio e enxofre afetou, negativamente, o desenvolvimento deste percevejo predador, prolongando seu est?gio imaturo. Em rela??o ao efeito da fitofagia nas plantas, verificou-se que P. nigrispinus n?o causa quaisquer danos ?s mudas de eucalipto, embora a extra??o de macronutrientes das plantas seja fundamental para estes insetos. O balan?o nutricional adequado das plantas ? essencial para obter sucesso em programas de controle biol?gico em que se utilizam predadores zoofit?fagos. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Produ??o Vegetal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2015. / ABSTRACT The search for explanations aimed at phytophagy stink bugs predators can provide subsidies enjoyed by biological control programs, making them more effective. Knowing that the macronutrients are involved in many physiological processes of insects, the aim of this study was to identify the effect of the absence of macronutrients in seedlings of eucalyptus in the development and food consumption Podisus nigrispinus as well, make sure to phytophagy, causes losses in plants. This study was conducted in a greenhouse and the treatments consisted of individual nitrogen omission, phosphorus, potassium, calcium, magnesium and sulfur in the nutrient solution, where the eucalyptus seedlings were being conducted. Pupae of Tenebrio molitor L. (Coleoptera: Teneobridae) were offered ad libitum to bedbugs? predators. The results of this experiment confirmed the importance of nutritional supplementation in plants for development and predation of the predator of Podisus nigrispinus. Feeding on plants deficient in nitrogen and magnesium reduced the predation efficiency of this zoophytophagous. And the calcium omission, magnesium and sulfur affected negatively on the development of this insect predators, extending their immature stage. Regarding on the effect of phytophagous in plants, the results confirmed that the predators do not cause any damage to seedlings of eucalyptus, although the extraction of plants are essential macronutrient for these insects. This work shows that the proper nutritional balance of the plants are essential for success in biological control programs that use zoophytophagous predators.
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Stomoxys calcitrans: Estabelecimento de col?nia e efeito de Metarhizium anisopliae sobre seus est?gios imaturos / Stomoxys calcitrans: colony establishment and effect of Metarhizium anisopliae in their immatures stages

Moraes, Ana Paula Rodrigues 27 February 2007 (has links)
Made available in DSpace on 2016-04-28T20:15:25Z (GMT). No. of bitstreams: 1 2007-Ana Paula Rodrigues Moraes.pdf: 1193160 bytes, checksum: 61d4adc426c0fdbeab75966756651ece (MD5) Previous issue date: 2007-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Stomoxys calcitrans is a hematophagous fly that is related to reduction in animal production due to the stress and dissemination of pathogens. The knowledge in plague control, using entomopathogenic fungi, has been growing year by year, but few works have been done to control S. calcitrans. The aims of this study were rearing a colony of S. calcitrans to make a bioassay of microbial control, verify the pathogenic potential of Metarhizium anisopliae in eggs, larvae and pupae and access three methods of egg s exposition to the studied fungi. After methodologies adaptation of rearing S. calcitrans that are described in literature, the colony was maintained in the Laborat?rio de D?pteros Hemat?fagos - UFRRJ. The bioassays were done in the Laborat?rio de Controle Microbiano de Atr?podes de Import?ncia Veterin?ria - UFRRJ, using the strain 959 of M. anisopliae in a water suspension. In the first method, eggs were immersed in the suspension (2.2 ? 108 conidia /ml and dilutions) and transferred to Petri plates with hydrophilic cotton. In the second methods (2.3 ? 108 conidia/ml and dilutions), eggs were immersed and transferred to the larval rearding medium. In the third method (2.1 ? 108 conidia/ml and dilutions), eggs were put in the larval rearding medium surface and one aliquot of fungus suspension were added. Larvae with nine days of development were immersed in suspension (2.1 ? 108 conidia/ml and dilutions) and after that were transferred to the larval rearding medium while pupae with 16 days of development were immersed in suspension (2.3 ? 108 conidia/ml and dilutions) and were held in Petri plates. Experiments were carried out with 107, 106 and 105 concentrations of conidia/ml and positive and negative controls. The mortality was assessed on the 10th day after fungus exposure. It was verified high mortality in all treated and control groups in the first method of eggs exposure. In the other two methods the eggs exposed to 108 conidia/ml concentration were 100% unviable. In the second method the eggs was 92.5% unviable and the third method was 53.33% both in 107 conidia/ml concentration. The subsequent concentrations did not show significant difference when compared to controls, neither between two methods of fungi exposure. Larvae and pupae exposure did not show statistic difference between treated and controls groups. Metarhizium anisopliae suspensions in high concentrations were capable to make unfeasible eggs of S. calcitrans; however, it did not happen in larvae and pupae experiments, in which fungi did not prevent their development. The second method was the best one to control immature forms of S. calcitrans but the third one resembles the nature conditions, in which has significant result only in the highest fungi concentration. Experimental methodologies that simulate nature conditions may optimize the utilization of fungi in arthropods control, in a way that it does not modify its action when applied in the field. / Stomoxys calcitrans ? uma mosca hemat?faga associada ? redu??o da produtividade, devido ao estresse causado aos animais e a dissemina??o de pat?genos. O conhecimento do controle de pragas, utilizando fungos entomopatog?nicos, vem crescendo no decorrer dos anos, por?m poucos trabalhos t?m sido realizados visando o controle de S. calcitrans. O objetivo deste estudo foi estabelecer uma col?nia de S. calcitrans direcionada ao bioensaio de controle microbiano, verificar o potencial patog?nico de Metarhizium anisopliae sobre ovos, larvas e pupas, e ainda, avaliar tr?s m?todos de exposi??o dos ovos ao fungo estudado. Ap?s adapta??o das metodologias de cria??o de S. calcitrans descritas na literatura, a col?nia foi mantida no Laborat?rio de D?pteros Hemat?fagos UFRRJ. Os bioensaios foram realizados no Laborat?rio de Controle Microbiano de Artr?podes de Import?ncia Veterin?ria UFRRJ, onde foi utilizada a cepa 959 de M. anisopliae para elabora??o das suspens?es aquosas. No primeiro m?todo, os ovos foram imersos na suspens?o (2,2 x 108 con?dios/ml e dilui??es) e transferidos para placa de Petri contendo algod?o hidr?filo. No segundo m?todo (2,3 x 108 con?dios/ml e dilui??es) os ovos foram imersos e transferidos para o meio de desenvolvimento larval. No terceiro m?todo (2,1 x 108 con?dios/ml e dilui??es), os ovos foram depositados sobre a superf?cie do meio de desenvolvimento larval e uma al?quota da suspens?o f?ngica foi adicionada. As larvas com nove dias de desenvolvimento foram imersas na suspens?o (2,1 x 108 con?dios/ml e dilui??es) e transferidas para o meio de desenvolvimento larval, enquanto que as pupas de 16 dias de desenvolvimento foram imersas na suspens?o (2,3 x 108 con?dios/ml e dilui??es) e depositadas em placas de Petri. Foram utilizadas dilui??es na concentra??o de 107, 106 e 105 con?dios/ml e controle positivo e negativo. A mortalidade foi avaliada no d?cimo dia ap?s exposi??o, onde foi observado se ocorria mudan?a de est?gio. Foi verificada alta mortalidade em todos os grupos (tratados e controles) no primeiro m?todo de exposi??o de ovos. Nos outros dois m?todos, foram obtidos 100% de inviabilidade dos ovos expostos ? concentra??o de 108 con?dios/ml. No segundo m?todo, houve 92,5% de inviabilidade dos ovos, enquanto que no terceiro m?todo, a inviabilidade foi de 53,33%, ambos na concentra??o de 107 con?dios/ml. As concentra??es subseq?entes n?o apresentaram diferen?a significativa frente aos controles e nem entre os dois m?todos de exposi??o f?ngica. Nas exposi??es de larvas e pupas n?o ocorreu diferen?a estat?stica entre os tratamentos e os controles. Suspens?es de M. anisopliae em altas concentra??es s?o capazes de inviabilizar ovos de S. calcitrans, entretanto, isto n?o ocorreu em larvas e pupas, pois o fungo n?o impediu o desenvolvimento destes. O m?todo mais efetivo para o controle das formas imaturas de S.calcitrans foi o segundo m?todo, entretanto, o que mais se assemelha ?s condi??es naturais seria o terceiro, onde foi obtido resultado significativo apenas na maior concentra??o f?ngica. A ado??o de metodologias experimentais que simulem condi??es naturais pode otimizar a utiliza??o de fungos no controle de artr?podes, de forma que sua a??o n?o seja comprometida, quando aplicados no campo.
5

Avalia??o in vitro dos efeitos de Isaria farinosa, I. fumosorosea, Paecilomyces lilacinus e Lecanicillium lecanii sobre Boophilus microplus. / In vitro evaluation of the effects of Isaria farinosa, I. fumosorosea, Paecilomyces lilacinus and Lecanicillium lecanii toward Boophilus microplus.

Angelo, Isabele da Costa 26 February 2007 (has links)
Made available in DSpace on 2016-04-28T20:15:27Z (GMT). No. of bitstreams: 1 2007-Isabele da Costa Angelo.pdf: 1629986 bytes, checksum: 663b35706e830a57e185c3e0a68096c9 (MD5) Previous issue date: 2007-02-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Boophilus microplus is an ectoparasite which causes great economic losses around the world. The exclusive use of acaricides and the inadequate management have conducted the development of resistance in ticks populations, environmental and food contamination by acaricides and their residues. The use of entomopathogenic fungi to arthropods control has shown interesting responses. Several studies have proved the pathogenicity of fungi toward various ticks species. The aim of the present study was to evaluate the effect of Isaria farinosa, I. fumosorosea, Paecilomyces lilacinus and Lecanicillium lecanii toward engorged females, eggs and larvae of B. microplus tick. The isolates were cultured on malt extract medium at 25 ? 1?C and 80% of relative humidity for 15 days. Conidial suspensions were prepared in Tween 80 water solution (0.1% v/v). There were 4 treatment groups according to the following conidial concentrations: 105, 106, 107 and 108 conidia mL-1. The control group was made up of water and Tween 80 only (0.1% v/v). Treatment was based on immersion of the specimen in 1 mL of the conidial suspension. Each treatment group was made up of 10 repetitions. Changes in biology of engorged females, eggs viability and larvae mortality, were observed every 5 days up to the 20th day after treatment. The results have shown that L. lecanii changed biological parameters in engorged females through the reduction in posture period, nutritional rate, eggs production rate, and increase of incubation period. Isaria farinosa has shown reduction in posture period and in nutritional rate. Paecilomyces lilacinus was the unique isolate able to reduce the eclosion period of larvae from infected females. Isaria fumosorosea has reduced the nutritional rate and was the unique isolate that changed significantly in eclosion period when eggs were infected. Conidial concentrations reduced reproduction capacity of engorged females. The isolates of I. fumosorosea and L. lecanii have presented major potential to control B. microplus engorged females. All tested isolates have shown pathogenicity toward unfed larvae of B. microplus after in vitro infection. / Boophilus microplus ? um ectoparasito que causa grandes perdas na pecu?ria mundial. A utiliza??o exclusiva de produtos qu?micos no controle de carrapatos, associada ao manejo inadequado tem conduzido o desenvolvimento de popula??es de carrapatos resistentes, e a contamina??o dos produtos de origem animal e do ambiente pelos seus res?duos. O uso de fungos entomopatog?nicos no controle de artr?podes tem se tornado uma abordagem cada vez mais atrativa. In?meros trabalhos comprovam experimentalmente a patogenicidade dos fungos sobre diversas esp?cies de carrapatos. O objetivo do presente trabalho foi avaliar o efeito in vitro de Isaria farinosa, I. fumosorosea, Paecilomyces lilacinus e Lecanicillium lecanii sobre f?meas ingurgitadas, ovos e larvas de B. microplus. Os fungos foram repicados em meio de cultura extrato de malte, e mantidos em c?mara climatizada sob temperatura de 25?C ? 1 e umidade relativa de 80% por quinze dias. Suspens?es conidiais foram preparadas a partir do crescimento do fungo, cujos con?dios foram adicionados a solu??o de ?gua destilada e Tween 0,1%. As concentra??es 105, 106, 107 e 108 con?dios mL-1 formaram os grupos tratamento, juntamente com o grupo controle, constitu?do por ?gua destilada est?ril e Tween 0,1%. O tratamento constituiu-se de um mililitro da concentra??o conidial testada, e cada grupo foi formado por 10 repeti??es. Os par?metros de avalia??o observados para demonstrar o efeito dos fungos sobre o carrapato foram as altera??es biol?gicas de f?meas ingurgitadas, viabilidade de ovos tratados e percentual de mortalidade de larvas, acompanhado a cada cinco dias at? o 20? dia ap?s infec??o. Os resultados mostraram que L. lecanii causou altera??es nos par?metros biol?gicos de f?meas ingurgitadas, diminuindo o per?odo de postura, o ?ndice nutricional, o ?ndice de produ??o de ovos, e aumentando o per?odo de incuba??o. Isaria farinosa mostrou redu??o no per?odo da postura e no ?ndice nutricional de f?meas ingurgitadas. P. lilacinus foi o ?nico fungo capaz de reduzir o per?odo de eclos?o das larvas provenientes da infec??o de f?meas ingurgitadas. I. fumosorosea reduziu o ?ndice nutricional e foi o ?nico isolado que causou altera??o significativa no per?odo de eclos?o de larvas provenientes da infec??o de ovos. Diferentes concentra??es dos entomomopat?genos reduziram o potencial reprodutivo das f?meas ingurgitadas dos grupos tratados. Os isolados de I. fumosorosea e L. lecanii apresentaram maior potencial de controle para f?meas ingurgitadas de B. microplus. Todos os isolados testados mostraram patogenicidade para larvas n?o alimentadas de B. microplus ap?s infec??o in vitro.
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Avalia??o in vivo dos fungos entomopatog?nicos Metarhizium anisopliae e Beauveria bassiana associados ou n?o com deltametrina no controle de uma cepa do carrapato Boophilus microplus resistente a piretr?ides / In vivo evaluation of both entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana associated or not with deltamethrin to control a pyrethroid resitant strain of Boophilus microplus

Bahiense, Thiago Campanharo 26 February 2007 (has links)
Made available in DSpace on 2016-04-28T20:16:20Z (GMT). No. of bitstreams: 1 2007-Thiago Campanharo Bahiense.pdf: 1378749 bytes, checksum: ef9fdcee9b7f5b82132b5b8d53a3fbe5 (MD5) Previous issue date: 2007-02-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Boophilus microplus can cause several cattle pathology and thus economic losses. Moreover, this tick species is able to develop resistance to acaricides. The biological control is a great promise to control cattle ticks populations. The aim of the present study was to contribute to the development of a new complementary method to control B. microplus. Therefore, it has been evaluated the compatibility between deltamethrin and the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae toward B. microplus tick in a stable test. It was used a tick strain resistant to pyrethroid and calves were artificially infested. A total of 8 calves were held in individual housing and infested with B. microplus larvae every other day for 25 days. After that, 4 treatments were realized with 2 calves per group. The control group was tested receiving just water; the second group was tested receiving M. anisopliae in a 108 conidia ml-1 suspension; the third one was tested receiving a 25 ppm deltamethrin solution; and the forth group was tested receiving a solution containing a 108 conidia ml-1 M. anisopliae suspension associated with a 25 ppm deltamethrin solution. The mortality rate was 32.57% in the second group, 38.58% in the third group, and 30.92% in the forth one. The main changes in ticks biology were a decreased nutritional rate until the forth day after treatment. The deltamethrin group was an exception with a decreased rate until the second day after treatment. The production of eggs rate was also decreased until the second day after treatment in all 3 tested groups. Therefore, the association between deltamethrin and B. bassiana and M. anisopliae could be used for B. microplus integrated management. Based on compatibility between acaricide and biological agents, it could be possible the use of both simultaneously on infested cattle. Thus, it is possible to control ticks resistant to deltamethrin if an association with entomogenous fungi such as Beauveria bassiana and Metarhizium anisopliae is established. The best performance of these fungi, associated or not with deltamethrin, was observed for a short period after treatment. / O principal ectoparasito dos bovinos ? o carrapato Boophilus microplus, que tem como caracter?stica grande capacidade de resistir ?s adversidades do meio ambiente sendo causador de diversas patologias, severos preju?zos econ?micos aos produtores e possui grande capacidade de desenvolver resist?ncia a carrapaticidas qu?micos. As pesquisas realizadas utilizando controle alternativo de B. microplus t?m apontado que os fungos que naturalmente promovem a infec??o e at? a morte de diversas esp?cies e est?gios de vida de carrapatos s?o, promissores candidatos a esse papel de agente de controle microbiano. Com o objetivo de fornecer subs?dios que complementar?o o controle desta parasitose, avaliou-se in vivo a compatibilidade entre a deltametrina e os fungos Beauveria bassiana e Metarhizium anisopliae sobre cepa resistente de B. microplus a deltametrina, atrav?s do teste de est?bulo, com bovinos infestados artificialmente. Foram utilizados oito bezerros, para cada bioensaio, alojados individualmente, infestados em dias alternados com larvas de B. microplus por um per?odo de 25 dias. Ao t?rmino da infesta??o, foram realizados quatro tratamentos com dois animais por grupo. O grupo controle recebeu somente ?gua; o grupo tratado com fungo recebeu suspens?o conidial na concentra??o de 108 con?dios/ml; o grupo tratado com deltametrina recebeu solu??o na concentra??o de 25 ppm e o grupo tratado com associa??o recebeu suspens?o do fungo na concentra??o de 108 con?dios/ml com deltametrina a 25 ppm. Os resultados foram obtidos atrav?s da contagem di?ria dos carrapatos desprendidos do corpo dos animais, al?m da avalia??o dos par?metros biol?gicos das f?meas, as quais foram incubadas em c?mara climatizada. Realizaram-se dois bioensaios, sendo o primeiro com o fungo Metarhizium anisopliae, onde os percentuais m?dios de mortalidade foram: 32,57% para este fungo, 38,58 % para deltametrina e 30,92% para associa??o entre ambos. No segundo bioensaio foi utilizado o fungo Beauveria bassiana, onde os percentuais m?dios de mortalidade foram: 26,06% para este fungo, 64,30% para deltametrina e 70,64% para associa??o entre ambos. Quanto aos par?metros biol?gicos, as principais altera??es em ambos bioensaios foram a redu??o do ?ndice nutricional, principalmente at? o quarto dia ap?s tratamento, com exce??o da deltametrina que reduziu at? o segundo dia ap?s o tratamento. O ?ndice de produ??o de ovos s? foi reduzido at? o segundo dia ap?s tratamento para todos os grupos do primeiro bioensaio e somente no quarto dia ap?s o tratamento para o B. bassiana. Sendo assim, a associa??o entre deltametrina e os fungos testados pode ser utilizada como ferramenta para o manejo integrado de B. microplus, demonstrando compatibilidade in vivo entre acaricida qu?mico e bio l?gico, sendo poss?vel sua utiliza??o, simult?nea sobre bovinos infestados com B. microplus. O melhor desempenho desses fungos, associados ou n?o, foi observado em um per?odo curto ap?s o tratamento.
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Caracteriza??o e sele??o de isolados de Beauveria bassiana para o controle microbiano do carrapato Boophilus microplus / Characterization and selection of Beauveria bassiana isolates to microbiological control of Boophilus microplus tick

Fernandes, ?verton Kort Kamp 30 March 2007 (has links)
Made available in DSpace on 2016-04-28T20:16:21Z (GMT). No. of bitstreams: 1 2007-Everton Kort Kamp Fernandes.pdf: 2844359 bytes, checksum: a56e9f806dc8f1f0e7cc6b4641b4a78c (MD5) Previous issue date: 2007-03-30 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Beauveria bassiana is an entomopathogenic fungus used worldwide as a biological control agent of pest arthropods. The present study aimed at characterizing several isolates of B. bassiana and selects those with major potential to control ticks population and other arthropods under natural conditions. Thus, 53 isolates of B. bassiana and 7 isolates including 5 other Beauveria spp. and 1 Engyodontium albus (=Beauveria alba) were investigated. These isolates were originally from different regions of Brazil and other nations, and different hosts or substrate. Initially, all the isolates were morphologically analyzed and investigated about the potential of conidial production. Afterwards, experiments were conducted to investigate the genetic diversity of Brazilian isolates of B. bassiana, to evaluate the spatial distribution of genotypes, and to understand the genetic relationship among isolates of different species of Beauveria. Thus, all isolates were analyzed by isozymes electrophoresis, AFLP-PCR and sequencing of ITS1 and ITS2 regions. The results have permitted the taxonomic identification of isolates and the selection of those with major potential to conidial production. The results have also showed the detection of high genetic variability among B. bassiana isolates, indicating that this fungus represents a species aggregate. Furthermore, the genetic distance among isolates was directly related to the geographic distances. In addition, the isolates were evaluated to UV-B tolerance, heat tolerance, and cold activity. Heat and cold are two abiotic factors of the environment that restrict the use of entomopathogenic fungi as agents for biological control of insects. In addition, solar radiation, particularly the UV-B wavelength, is a major environmental factor that can negatively affect this entomopathogenic fungus in the field. High variability in UV-B tolerance was detected among the isolates after 2 h of radiation exposure. Conidial relative germination ranged from 0% to almost 80%, and a delayed germination was detected. Moreover, isolates of B. bassiana originating from lower latitudes tended to have lower UV-B tolerance than isolates from higher latitudes. A high variability in thermotolerance was also observed among the isolates after 2 h of heat exposure at 45 ?C, ranging from low, to medium, to high viability. The thermal death point of most resistant isolates was between 44 ?C and 45 ?C after 6 h of heat exposure. At cold temperatures most of the isolates grew at 5 ?C. An attempt to correlate the latitude of origin with thermotolerance or cold activity indicated that isolates from higher latitudes were more cold-active than isolates from nearer the equator. The present study also evaluated the virulence of the isolates toward Boophilus microplus larvae, and virulence ranged from very low to very high. It was observed that different populations of B. microplus may present different susceptibility to B. bassiana infection. Therefore, the present study has selected isolates of this entomopathogenic fungus with major effective potential to control B. microplus and other pest arthropods under environmental conditions. / Beauveria bassiana ? um fungo entomopatog?nico utilizado em v?rias partes do mundo como agente de controle biol?gico de artr?podes. Baseado nisso, o presente estudo buscou caracterizar diversos isolados de B. bassiana e selecionar aqueles com maior potencial para controlar popula??es de carrapatos e outros artr?podes em condi??es naturais. Neste sentido, 53 isolados de B. bassiana e mais sete isolados incluindo cinco esp?cies de Beauveria spp. e uma de Engyodontium albus (=Beauveria alba) foram estudadas. Esses isolados foram oriundos de v?rias regi?es geogr?ficas brasileiras e estrangeiras, e de diferentes hospedeiros ou substratos. Os isolados foram inicialmente avaliados morfologicamente e quanto ao potencial de produ??o de con?dios. Em seguida, buscou-se investigar a diversidade gen?tica dos isolados brasileiros de B. bassiana, avaliar a distribui??o espacial de gen?tipos, e conhecer a rela??o gen?tica entre os isolados das diferentes esp?cies. Para esta finalidade os isolados foram submetidos as t?cnicas de eletroforese de isoenzimas, AFLP-PCR e seq?enciamento de regi?es ITS1 e ITS2. Os resultados permitiram a confirma??o taxon?mica dos isolados e a sele??o daqueles com maior capacidade de produ??o de con?dios. Permitiram tamb?m detectar grande variabilidade gen?tica entre os isolados de B. bassiana indicando que este fungo representa um agregado de esp?cies. Al?m disso, a dist?ncia gen?tica detectada entre os isolados foi fortemente relacionada a dist?ncia geogr?fica de origem dos isolados. Posteriormente, os isolados tamb?m foram avaliados quanto a toler?ncia ? radia??o UV-B, toler?ncia ao calor e atividade ao frio. Tanto o calor quanto o frio s?o fatores abi?ticos ambientais que restringem o uso de fungos como agentes de biocontrole, enquanto a radia??o solar, particularmente UV-B, ? o maior fator ambiental que pode afetar negativamente a a??o deste entomopat?geno. Neste sentido, foi observada alta variabilidade entre os isolados analisados em rela??o ? toler?ncia a UV-B ap?s duas horas de exposi??o ? radia??o, obtendose percentuais de germina??o de con?dios que variaram entre 0% e aproximadamente 80%. Al?m disso, foi observado atraso na germina??o dos con?dios, e revelado que os isolados originados de baixas latitudes tenderam a apresentar menor toler?nc ia a UV-B do que aqueles provenientes de latitudes mais elevadas. Foi tamb?m encontrada alta variabilidade em termotoler?ncia dos con?dios ap?s duas horas de exposi??o a 45 ?C, variando entre baixa, m?dia e alta viabilidade. Os con?dios mais resistentes n?o toleraram exposi??o entre 44 ?C e 45 ?C por seis horas. Quando submetidos a 5 ?C a maioria dos isolados apresentaram atividade. Os isolados de B. bassiana obtidos de latitudes mais elevadas foram mais ativos ao frio do que aqueles originados pr?ximos ? linha do equador. Al?m disso, os isolados tamb?m foram avaliados quanto ao potencial virulento sobre larvas de Boophilus microplus, e neste sentido foi poss?vel diferenci?-los em baixa, m?dia e alta virul?ncia. Foi detectado tamb?m que diferentes popula??es desta esp?cie de carrapato podem manifestar diferentes n?veis de suscetibilidade ? infec??o por B. bassiana. Neste sentido, o presente estudo permitiu selecionar isolados deste entomopat?geno com maior potencial para o controle biol?gico de B. microplus e outros artr?podes em condi??es de campo.
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Identifica??o e quantifica??o de hem?citos de f?meas ingurgitadas de Boophilus microplus inoculados com fungos Metarhizium anisopliae, Beauveria bassiana, Penicillium corylophilum e Fusarium oxysporum. / Identification and quantification of hemocytes obtained from engorged females of Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) inoculated with Metarhizium anisopliae, Beauveria bassiana, Penicillium corylophilum and Fusarium oxysporum

Silva, Sandra Borges da 24 February 2006 (has links)
Made available in DSpace on 2016-04-28T20:16:27Z (GMT). No. of bitstreams: 1 2006-Sandra Borges da Silva.pdf: 1572214 bytes, checksum: 1980363c45abbde9170747be40d0635f (MD5) Previous issue date: 2006-02-24 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Biological control of the tick Boophilus microplus with Metarhizium anisopliae and Beauveria bassiana has been evaluated by several researchers. The selection of specimens resistant to chemical products is a mechanism used by arthropods for survival and maintenance in the environment. It is already known how fungi penetrate the host and their pathogenic potential at different phases of their biological cycle, however, ticks immune response against these agents need more detailed studies. The present work had as objectives: to identify and quantify the cellular types involved in the cellular response of B. microplus inoculated with entomopathogenic (M. anisopliae and B. bassiana) and non-entomopathogenic fungi (Penicillium corylophilum and Fusarium oxysporum). In this study 60 engorged females of B. microplus were used, representing six treatment groups with 10 specimens each. Ticks were inoculated with aqueous suspension of conidia. There were two control groups: in the first one ticks were inoculated with Tween 80 0,1% aqueous solution (negative control), in the second one ticks were not inoculated (testimony group). Fungi suspension or Tween 80 solution were applied in the back part of the idiosoma of the tick. The hemolinfa samples were collected during the whole period of life of the specimens, beginning 24 hours after inoculation. The haemolymph samples were fixed in methanol and stained with Giemsa. In all studied periods, cells like pro-hemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids were observed in the specimens of all groups. Prohemocytes, plasmatocytes and spherulocytes were the most numerous cells observed in the hemolinfa. Hemocytes were absent in the group inoculated with B. bassiana 72 hours after inoculation. After this period, specimens inoculated with entomopathogenic fungi were dead. The absence of cells suggests the immune-suppressor effect of the fungi on this tick species. It was not observed when ticks were treated with M. anisopliae due to accentuated mortality caused by this fungus species. The nonentomopathogenic fungi did not affect the life cycle of this tick, being quickly eliminated of the organism. It was suggested based on the absence of conidia during sampling. / O controle biol?gico do carrapato Boophilus microplus com a utiliza??o de Metarhizium anisopliae e Beauveria bassiana tem sido avaliado por diversos pesquisadores. A sele??o de cepas resistentes ? um mecanismo adotado pelos artr?podes para sobreviv?ncia da esp?cie e sua manuten??o no ambiente. Apesar de estar esclarecida a forma como o fungo penetra no hospedeiro e comprovada sua patogenicidade sobre diferentes fases de seu ciclo biol?gico, a sua resposta imune frente a estes agentes agressores necessita de maiores estudos. O presente trabalho teve como objetivo: identificar e quantificar os tipos celulares envolvidos na resposta imune celular de B. microplus inoculados com fungos entomopatog?nicos n?o entomopatog?nicos. Na experimenta??o foram utilizados 60 f?meas ingurgitadas de Boophilus microplus, representando seis tratamentos cada um contendo 10 esp?cimes. Para os grupos inoculados com fungos entomopatog?nicos foi utilizada a suspens?o aquosa dos fungos Metarhizium anisopliae (isolado Ma 959) e Beauveria bassiana (isolado Bb 986). Nos tratamentos com fungos n?o entomopatog?nicos, Penicillium corylophilum e Fusarium oxysporum, foram formados ainda um grupo testemunha (n?o recebeu inocula??o) e um grupo inoculado com solu??o de tween 80 a 0,1% em ?gua destilada est?ril, considerado controle negativo. Nos grupos dos tratamentos com suspens?o f?ngica ou solu??o a inocula??o foi feita na regi?o posterior do idiossoma do carrapato. As coletas de hemolinfa foram realizadas durante todo o per?odo de vida dos esp?cimes, tendo inicio 24 horas ap?s inocula??o. As amostras de hemolinfa foram fixadas com metanol e coradas com Giemsa. Em todos os per?odos estudados, tanto nos esp?cimes inoculados com fungos como nos controles, foram observados pr?-hem?citos, plasmat?citos, granul?citos, esferul?citos e oenocit?ides. Pr?-hem?citos, plasmat?citos e esferul?citos foram ?s c?lulas mais numerosas na hemolinfa. Foi observada a aus?ncia de hem?citos no grupo inoculado com B. bassiana 72 horas ap?s a inocula??o, como tamb?m a morte dos esp?cimes inoculados com fungos entomopatog?nicos a partir deste per?odo. A aus?ncia de c?lulas evid?ncia o efeito imunossupressor do fungo sobre os carrapatos estudados. Caracter?stica n?o observada para Metarhizium anisopliae, mesmo tendo provocado acentuada mortalidade dos carrapatos. Os fungos n?o entomopatog?nicos n?o afetaram de forma significativa o ciclo de vida destes carrapatos, sendo rapidamente eliminados do organismo.
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Avalia??o dos Perfis Prot?ico e Lip?dico na Resposta de Rhipicephalus Microplus ? Infec??o com Fungos. / Evaluation of protein and lipid profile in response of Rhipicephalus microplus to infection by fungi.

Angelo, Isabele da Costa 03 March 2011 (has links)
Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-06-09T16:27:04Z No. of bitstreams: 1 ISABELE DA COSTA ANGELO.pdf: 4140152 bytes, checksum: bbd35489ea24b040be77df36cd797987 (MD5) / Made available in DSpace on 2016-06-09T16:27:32Z (GMT). No. of bitstreams: 1 ISABELE DA COSTA ANGELO.pdf: 4140152 bytes, checksum: bbd35489ea24b040be77df36cd797987 (MD5) Previous issue date: 2011-03-03 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The study evaluated the protein and lipid profiles of Rhipicephalus microplus engorged females after infection by Metarhizium anisopliae, Beauveria bassiana and Fusarium oxysporum. The treatments were immersion or inoculation of conidial suspension in R. microplus. The hemolymph was collected 24 and 48 hours after treatment. The cell-free hemolymph was separated of hemocytes by centrifugation and hemocytes resuspended in phosphate buffer pH 7.2. The amount of total protein was determined in both fractions of hemolymph and hemocytes were quantified. The cell-free hemolymph was filtered through a 100 kDa and 10 kDa membranes, and analyzed by electrophoresis and liquid chromatography (HPLC). The proteome of cell-free hemolymph (treatment by injection) was evaluated by 2DPAGE. Changes were observed in amount total protein and the amount of hemocytes, but no difference was observed in the electrophoretic profile (1D-PAGE) of the cell-free hemolymph. In haemocytes, the entomopathogens reduced the amount of serpins, while F. oxysporum caused increased. In 2D-PAGE variations were observed in both expression and presence/absence of protein between the groups. The cell-free hemolymph antimicrobial activity was tested against Escherichia coli and Staphylococcus aureus and against the fungus used in the treatment of engorged ticks. The hemolymph collected 48 hours after the immersion treatment with B. bassiana apresented activity anti-B. bassiana with 48 hours of evaluation. This hemolymph was subjected to Superose column to HPLC and peak was collected and analyzed on the analytical column C18. The fractions were collected from the C18 and its apresented activity anti-B. bassiana, but showed no activity against Candida albicans. These fractions were analyzed by Maldi-Tof and most of them had in common an ion with m/z 1,119.5; however, other ions may be involved with this activity antimicrobial. The lipids present in cell-free hemolymph, in the hemocyte and fat body were extracted and analyzed by thin layer chromatography (TLC) or HPTLC for neutral lipids and phospholipids. The classes of neutral lipids in the cell-free hemolymph were cholesterol ester, cholesterol (CHO) and fatty acids (FA), which have varied depending on the fungus used, type of treatment and observation time. Phospholipids found were phosphatidylcholine and phosphatidylethanolamine, its were not significantly altered after the fungal infection. In hemocytes, the same classes of lipids were found and B. bassiana modified phospholipids, while M. anisopliae s.l. altered FA and CHO. The fat body showed, in addition to these classes of neutral lipids, the triglycerides, which increased significantly 48 hours after inoculation with M. anisopliae s.l. The lipase activity in fat body was measured and it was demonstrated that increased activity 48 hours after inoculation, mainly in the group inoculated with Metarhizium. Therefore, the results showed alterations related to the proteins expression in the hemocytes and the cell-free hemolymph after inoculation with fungi, immunosuppression of hemocytes and antimicrobial peptides induction after infection with B. bassiana, besides changes in the lipid profile of R. microplus after infection. However, further studies are necessary to understand these changes. / O trabalho avaliou os perfis prot?ico e lip?dico de f?meas ingurgitadas de Rhipicephalus microplus ap?s infec??o com Metarhizium anisopliae s.l., Beauveria bassiana ou Fusarium oxysporum. Os tratamentos foram imers?o ou inocula??o da suspens?o conidial em R. microplus. A hemolinfa foi coletada 24 e 48 horas ap?s os tratamentos. O plasma foi separado dos hem?citos por centrifuga??o e os hem?citos resuspensos em tamp?o fosfato pH 7,2. A concentra??o de prote?na total foi determinada em ambas as fra??es da hemolinfa e os hem?citos quantificados. O plasma da hemolinfa foi filtrado em membrana de 100 kDa e 10 kDa, sendo analisados por eletroforese e cromatografia l?quida de alta efici?ncia (CLAE). O proteoma do plasma da hemolinfa (tratamento por inocula??o) foi avaliado por gel 2D. Foram observadas varia??es na quantidade de prote?na total de ambas as fra??es da hemolinfa, na quantidade de hem?citos bem como na intensidade de prote?nas/pept?deos expressos no plasma da hemolinfa. Nos hem?citos, os entomopat?genos reduziram a quantidade de serpinas, enquanto F. oxysporum causou aumento. No gel 2D foram observadas varia??es na express?o bem como na aus?ncia/presen?a de prote?nas entre os grupos. O plasma da hemolinfa teve sua atividade antimicrobiana testada contra Escherichia coli e Staphylococcus aureus e contra o fungo utilizado no tratamento das f?meas ingurgitadas. A hemolinfa coletada 48 horas ap?s o tratamento por imers?o com B. bassiana apresentou atividade anti-B. bassiana com 48 horas de avalia??o. Esta hemolinfa foi submetida ? coluna Superose de CLAE e o pico coletado analisado na coluna anal?tica C18. As fra??es coletadas da C18 apresentaram atividade anti-B. bassiana, por?m n?o apresentaram atividade contra Candida albicans. Essas fra??es foram analisadas por Maldi-Tof e a maioria delas apresentou um ?on com raz?o m/z 1.119,5; no entanto, outros ?ons podem estar envolvidos com essa atividade antimicrobiana. Os lip?deos presentes no plasma da hemolinfa, nos hem?citos e no corpo gorduroso foram extra?dos e analisados por cromatografia em camada delgada (CCD) ou CCD de alta performance para lip?deos neutros e fosfolip?deos. As classes de lip?deos neutros encontradas no plasma da hemolinfa foram colesterol-?ster, colesterol (CHO) e ?cidos graxos (AG), que sofreram altera??es em fun??o do fungo utilizado, tipo de tratamento e tempo de observa??o. Os fosfolip?deos encontrados foram fosfatidilcolina e fosfatidiletanolamina, que n?o foram significativamente alterados ap?s a infec??o f?ngica. Nos hem?citos, as mesmas classes de lip?deos foram encontradas e B. bassiana alterou os fosfolip?deos, enquanto M. anisopliae s.l. alterou os AG e CHO. O corpo gorduroso apresentou, al?m destas classes de lip?deos neutros, o triacilglicerol, que aumentou significativamente 48 horas ap?s a inocula??o com M. anisopliae s.l. A atividade lipase no corpo gorduroso foi mensurada, sendo evidenciado um aumento 48 horas ap?s a inocula??o, principalmente no grupo inoculado com Metarhizium. Portanto, os resultados demonstraram altera??es na express?o de prote?nas no plasma da hemolinfa e nos hem?citos ap?s inocula??o com os fungos, imunossupress?o dos hem?citos, indu??o de pept?deos com atividade antimicrobiana ap?s infec??o com B. bassiana, al?m de altera??es no perfil lip?dico de R. microplus ap?s infec??o. No entanto, maiores estudos s?o necess?rios para o entendimento dessas altera??es.
10

Avalia??o experimental de Lecanicillium lecanii no controle biol?gico de Stomoxys calcitrans / Experimental evaluation of Lecanicillium lecanii in biological control of Stomoxys calcitrans

ALVES, Paula Sant?ana 26 February 2010 (has links)
Submitted by Leticia Schettini (leticia@ufrrj.br) on 2016-08-29T16:13:08Z No. of bitstreams: 1 2010 - Paula Sant?ana Alves.pdf: 1049254 bytes, checksum: 295fd608eecdae86916f71cbebfc389e (MD5) / Made available in DSpace on 2016-08-29T16:13:08Z (GMT). No. of bitstreams: 1 2010 - Paula Sant?ana Alves.pdf: 1049254 bytes, checksum: 295fd608eecdae86916f71cbebfc389e (MD5) Previous issue date: 2016-02-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPQ / Stomoxys calcitrans is a hematophagous fly of which parasitism causes economic losses related to the reduction of milk production and weight gain. Due to the high level of resistance of the stable fly to most of chemical insecticides, it is necessary to search for new alternatives for their control, and one of them would be the use of entomopathogenic fungi. However, few researches related to microbial control of this fly were carried out. The objective of this study was to assess the ability of the Lecanicillium lecanii fungus in to make unfeasible eggs, larvae and pupae of the stable fly under laboratory conditions and verify the formation of inhibition zone in agar solid due to the presence of substances in the mucus and total homogenized larvae not previously exposed and sensitized to the fungus. The S. calcitrans colony was maintained in the Laborat?rio de D?pteros Hemat?fagos, UFRRJ. The tests were performed in the Laborat?rio de Controle Microbiano de Artr?podes, UFRRJ, where the strain CG 420 of L. lecanii was used to prepare the water suspensions. In assays with eggs, larvae and pupae of S. calcitrans, fungal suspensions were used at 2x108, 2x107, 2x106, 2x105 con.mL-1 concentrations, besides the positive control. In assays with eggs, they were immersed in different fungal suspension and transferred to Petri dishes with paper filter, which were added larval rearding medium and its respective suspension. Mortality was assessed five days after exposure, where it was observed whether stage change occurred. There was no significant difference in the hatching of eggs exposed to different fungal concentrations against the control. Larvae were subjected to the same treatment of eggs, and mortality was assessed ten to fifteen days after exposure. There was no significant difference in mortality of larvae exposed to the fungus when compared with the control group, however, the fungus L. lecanii was significantly able to reduce adult emergence from larvae exposed to the highest tested concentration and there was a percentage of emergence of 25,45% and 43,64% compared to 63,64% and 90,91% of the control group. The pupae were also immersed in different fungal suspensions, and transferred to Petri dishes with paper filter, added to its respective suspension. Mortality was assessed ten days after exposure. There was no significant difference in mortality of pupae exposed to different fungi concentrations. Filter paper discs were soaked in mucus samples and total homogenized from larvae of S. calcitrans not exposed and previously sensitized to the fungus L. lecanii and placed in petri dishes sown with the same fungus. It was not observed the formation of inhibition zone around the discs. The L. lecanii fungus was unable to make unfeasible eggs, larvae or pupae of S. calcitrans in tested fungal concentrations, however, was significantly able to reduce the emergence of adults when their third larvae stage were exposed to 2x108 con.mL-1 concentration. / Stomoxys calcitrans ? um d?ptero hemat?fago cujo parasitismo ocasiona perdas econ?micas relacionadas ? redu??o da produ??o leiteira e ganho de peso, al?m de estar associado ? dissemina??o e transmiss?o de agentes patog?nicos. Devido ao elevado grau de resist?ncia da mosca dos est?bulos a uma diversidade de inseticidas qu?micos, torna-se necess?rio a busca por alternativas para o seu controle, e uma delas seria a utiliza??o de fungos entomopatog?nicos. Todavia, poucas pesquisas relacionadas ao controle microbiano desta mosca foram realizadas. O objetivo deste estudo foi verificar a capacidade do fungo Lecanicillium lecanii em inviabilizar ovos, larvas e pupas de S. calcitrans em condi??es laboratoriais e verificar a forma??o de halo de inibi??o em meio de cultivo s?lido, em decorr?ncia de subst?ncias presentes no muco e no macerado total de larvas n?o expostas e previamente sensibilizadas ao fungo. A col?nia de S. calcitrans foi mantida no Laborat?rio de D?pteros Hemat?fagos, UFRRJ. Os ensaios foram realizados no Laborat?rio de Controle Microbiano de Artr?podes, UFRRJ. Nos ensaios foram utilizadas suspens?es aquosas do isolado CG 420 de L. lecanii nas concentra??es de 2x108, 2x107, 2x106, 2x105 con.mL-1, al?m do controle positivo. Nos ensaios com ovos, estes foram imersos nas suspens?es f?ngicas, e transferidos para placas de Petri com papel filtro, meio de desenvolvimento larval e 3 mL de sua respectiva suspens?o. A mortalidade foi avaliada cinco dias ap?s a exposi??o. N?o foi observada diferen?a significativa na eclos?o dos ovos tratados frente ao controle. As larvas foram submetidas ao mesmo tratamento dos ovos, e a mortalidade foi avaliada dez e quinze dias ap?s a exposi??o. N?o foi observada diferen?a significativa na mortalidade de larvas expostas ao fungo quando comparados com o grupo controle, entretanto, o fungo L. lecanii reduziu significativamente a emerg?ncia de adultos provenientes das larvas expostas ? concentra??o m?xima testada (2x108 con.mL-1) sendo observado um percentual de emerg?ncia de 25,45% e 41,82 %, em compara??o com 63,64% e 90,91% do grupo controle. As pupas tamb?m foram imersas nas suspens?es f?ngicas, e transferidas para placas de Petri com papel filtro e sua respectiva suspens?o. A mortalidade foi avaliada dez dias ap?s a exposi??o. N?o foi observada diferen?a significativa na mortalidade das pupas expostas ?s diferentes concentra??es f?ngicas. Discos de papel filtro foram embebidos em amostras de muco e macerado total proveniente de larvas de S. calcitrans n?o expostas e previamente sensibilizadas ao fungo L. lecanii e colocados em placas de Petri semeadas com o mesmo fungo (1x108 con.mL-1). N?o foi verificada a forma??o de halo de inibi??o ao redor dos discos. O fungo L. lecanii foi incapaz de inviabilizar ovos, larvas ou pupas de S. calcitrans nas concentra??es f?ngicas testadas, no entanto, reduziu significativamente a emerg?ncia de adultos quando suas larvas de terceiro est?dio foram expostas a concentra??o 2x108 con.mL-1.

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