Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)

Detalhes bibliográficos
Autor(a) principal: GACHOKA, Kennedy Kiriira
Data de Publicação: 2010
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tde/1087
Resumo: Amblyomma cajennense, commonly known as Cayenne tick, is a three-host tick widely distributed in Brazil and in the rest of Latin American countries. Owing to its low host specificity, it plays an important role in the transmission of pathogens in animals and man. Chemical communication is a vital component in tick biology whereby pheromones are largely responsible in the regulation of mating, recognition of sexual partner and copulation. By understanding the behaviour of A. cajennense and the pheromones involved, there exists practical possibility for the use of these substances as an additional method of control thereby reducing the use of acaricides. Based on this, the aims of this work were to quantify and elucidate behaviours induced by pheromones emitted by females A. cajennense as well as identify and characterize qualitatively and biologically the pheromone produced by males. Engorged females were collected from naturally-infested horses for the establishment of laboratory colonies for use in this study. The pheromones were obtained by extraction in hexane and sonication by ultra-sound of fed males and females. The female sex pheromone, 2,6-dichlorophenol (2,6-DCP), was quantified by GC/MS using internal standard and standard addition methods through construction of calibration curves using 5-bromine-4-hydroxy-3-methoxybenzaldehyde and 2,6-DCP, respectively. The male extract was analysed chemically by GC/MS in comparison with standards. As concerns biological tests, glass beads were treated with two femaleequivalents of the female pheromone and 2,6-DCP in an equivalent concentration and exposed to males to observe the induced behaviour(s). Attraction to the male pheromone was evaluated by release of individual unfed, virgin females to three concentrations (1, 5 and 10 male-equivalents) of the extract in olfactometer bioassays. Orientations towards the odour source were analysed by Kruskal-Wallis and displacement angles by Circular Statistics. Aggregation was evaluated by releasing the tick stages in Petri dish bioassays shortly followed by introduction of filter paper strips impregnated with 10 male-equivalents of the extract and recording tick responses on camera. Data analysis was by Chi-square test. Tests for attachment were carried out in vivo in rabbits, Oryctolagus cuniculus, and the released ticks observed in determined intervals between 20min and 24hrs. The data was analysed by Wilcoxon signed-ranked test and Friedman s repeated measures ANOVA. The results indicated that each female emits 2.15 ng ml-1 of pheromone on average. The male extract was positive for benzoic acid, nonanoic acid, salicylic acid, benzaldehyde, 2,6-DCP, limonene and methyl salicylate. The female extract and 2,6- DCP stimulated mounting in up to 80% and 73% of all cases, respectively, confirming that 2,6-DCP alone mediates mounting behaviour in A. cajennense. There was no statistically significant attraction or aggregation in males or females induced by the male pheromone. However, significantly faster and greater attachment was recorded in both. The behavioural responses corresponded with the pheromone s composition. It was concluded that the male pheromone promotes neither attraction nor aggregation but is responsible for faster fixation of A. cajennense adults. The pheromone also contains different proportions and differs from other Amblyomma species as far as the presence or absence of certain components is concerned. The results serve as a first step in advanced studies aimed at development of novel strategies for the management of this species using these pheromones.
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spelling BORGES, Lígia Miranda Ferreirahttp://lattes.cnpq.br/2591106734434348FERRI, Pedro Henriquehttp://lattes.cnpq.br/2129799749473005SILVA, Andrea Caetano dahttp://lattes.cnpq.br/1432918585717355http://lattes.cnpq.br/9231444052376453GACHOKA, Kennedy Kiriira2014-07-29T15:13:38Z2010-10-132010-08-13GACHOKA, Kennedy Kiriira. Chemical and biological pheromone of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae). 2010. 94 f. Tese (Doutorado em Ciências Agrárias) - Universidade Federal de Goiás, Goiânia, 2010.http://repositorio.bc.ufg.br/tede/handle/tde/1087Amblyomma cajennense, commonly known as Cayenne tick, is a three-host tick widely distributed in Brazil and in the rest of Latin American countries. Owing to its low host specificity, it plays an important role in the transmission of pathogens in animals and man. Chemical communication is a vital component in tick biology whereby pheromones are largely responsible in the regulation of mating, recognition of sexual partner and copulation. By understanding the behaviour of A. cajennense and the pheromones involved, there exists practical possibility for the use of these substances as an additional method of control thereby reducing the use of acaricides. Based on this, the aims of this work were to quantify and elucidate behaviours induced by pheromones emitted by females A. cajennense as well as identify and characterize qualitatively and biologically the pheromone produced by males. Engorged females were collected from naturally-infested horses for the establishment of laboratory colonies for use in this study. The pheromones were obtained by extraction in hexane and sonication by ultra-sound of fed males and females. The female sex pheromone, 2,6-dichlorophenol (2,6-DCP), was quantified by GC/MS using internal standard and standard addition methods through construction of calibration curves using 5-bromine-4-hydroxy-3-methoxybenzaldehyde and 2,6-DCP, respectively. The male extract was analysed chemically by GC/MS in comparison with standards. As concerns biological tests, glass beads were treated with two femaleequivalents of the female pheromone and 2,6-DCP in an equivalent concentration and exposed to males to observe the induced behaviour(s). Attraction to the male pheromone was evaluated by release of individual unfed, virgin females to three concentrations (1, 5 and 10 male-equivalents) of the extract in olfactometer bioassays. Orientations towards the odour source were analysed by Kruskal-Wallis and displacement angles by Circular Statistics. Aggregation was evaluated by releasing the tick stages in Petri dish bioassays shortly followed by introduction of filter paper strips impregnated with 10 male-equivalents of the extract and recording tick responses on camera. Data analysis was by Chi-square test. Tests for attachment were carried out in vivo in rabbits, Oryctolagus cuniculus, and the released ticks observed in determined intervals between 20min and 24hrs. The data was analysed by Wilcoxon signed-ranked test and Friedman s repeated measures ANOVA. The results indicated that each female emits 2.15 ng ml-1 of pheromone on average. The male extract was positive for benzoic acid, nonanoic acid, salicylic acid, benzaldehyde, 2,6-DCP, limonene and methyl salicylate. The female extract and 2,6- DCP stimulated mounting in up to 80% and 73% of all cases, respectively, confirming that 2,6-DCP alone mediates mounting behaviour in A. cajennense. There was no statistically significant attraction or aggregation in males or females induced by the male pheromone. However, significantly faster and greater attachment was recorded in both. The behavioural responses corresponded with the pheromone s composition. It was concluded that the male pheromone promotes neither attraction nor aggregation but is responsible for faster fixation of A. cajennense adults. The pheromone also contains different proportions and differs from other Amblyomma species as far as the presence or absence of certain components is concerned. The results serve as a first step in advanced studies aimed at development of novel strategies for the management of this species using these pheromones.Amblyomma cajennense, vulgarmente conhecido como carrapato-estrela, é um carrapato trioxeno que está amplamente difundido no território brasileiro e nos demais países da América Latina. Dada a sua baixa especificidade, assume um importante papel na transmissão de patógenos entre os animais e o homem. A comunicação química é um componente vital na biologia dos carrapatos, onde feromônios são majoritariamente responsáveis na regulação da busca pelo parceiro e a cópula. Compreendendo o comportamento de A. cajennense e os feromônios envolvidos, existem possibilidades práticas do uso destas substâncias como uma forma alternativa e satisfatória de controlálos minimizando o uso de acaricidas. Diante disso, procurou-se neste trabalho, quantificar e elucidar comportamentos instigados por feromônio produzido por fêmeas bem como identificar e caracterizar qualitativa e biologicamente feromônio de machos. Fêmeas ingurgitadas de A. cajennense foram colhidas de eqüinos naturalmente infestados para estabelecimento de colônias no laboratório. Os feromônios foram obtidos por extração e sonicação em hexano de machos ou fêmeas de carrapatos alimentados. Quantificou-se o feromônio sexual, o 2,6-diclorofenol (2,6-DCF), em fêmeas pelas técnicas de padrão interno, com 5-bromo-4-hidróxi-3-metóxi-benzaldeido como referência interna, e adição padrão por meio de cromatografia gasosa acoplada à espectrometria de massas (CG/EM) através de construção de curvas de calibração. O extrato dos machos foi analisado quimicamente por CG/EM diante de padrões sintéticos. No que concerne aos testes biológicos, pérolas de vidro foram tratadas com dois equivalentes-fêmea de amostra de feromônio extraído de fêmeas e com 2,6-DCF em uma dose equivalente e expostas aos machos para observar comportamento(s) provocado(s). A atração diante do feromônio de machos foi avaliada pela liberação individual de fêmeas, machos e ninfas virgens, não alimentadas, frente a três concentrações (1, 5 e 10 equivalentes-macho) do extrato de machos, em um olfatômetro. As orientações à fonte de odor foram analisadas por teste de Kruskal-Wallis e os ângulos por Estatística Circular. A agregação foi avaliada pela aplicação de 10 equivalentes-macho do extrato em arenas de placas de Petri e os dados analisados por Chi-quadrado. Testes de fixação foram feitos in vivo em coelhos, Oryctolagus cuniculus, onde os carrapatos foram liberados e observados em intervalos determinados entre 20min e 24hs e suas respostas analisadas por testes de Wilcoxon e ANOVA Friedman. Os resultados mostraram que uma fêmea libera em média 2,15 ng mL-1 de feromônio. O extrato de machos apresentou ácido benzóico, ácido nonanóico, ácido salicílico, benzaldeido, 2,6-DCF, limoneno e salicilato de metila. O extrato de fêmeas e o 2,6-DCF sintético estimularam a monta em até 73% de casos, respectivamente, confirmando que o composto é responsável por mediar à monta em A. cajennense. Não houve atração nem agregação estatisticamente maior dos machos e fêmeas diante do feromônio procedente de machos. Entretanto, uma fixação significativamente maior e mais rápida foi registrada dentro de 24 h em ambos os sexos. Os achados de respostas biológicas corresponderam com a composição química de feromônio de machos. Concluiu-se que o feromônio de machos não promove atração nem agregação, mas é responsável pela fixação de adultos de A. cajennense. O feromônio também possui diferenças na proporção e na presença de determinados compostos descritos em outras espécies do gênero Amblyomma. Os resultados estão discutidos como um ponto de partida para estudos avançados visando o desenvolvimento de novas estratégias de controle e de manejo desta espécie utilizando esses feromônios.Made available in DSpace on 2014-07-29T15:13:38Z (GMT). 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dc.title.por.fl_str_mv Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
dc.title.alternative.eng.fl_str_mv Chemical and biological pheromone of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
title Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
spellingShingle Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
GACHOKA, Kennedy Kiriira
carrapato-estrela
comunicação química
feromônios
semioquímicos
Cayenne tick
chemical communication
pheromones
semiochemicals
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
title_short Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
title_full Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
title_fullStr Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
title_full_unstemmed Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
title_sort Caracterização química e biológica de feromônios de Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae)
author GACHOKA, Kennedy Kiriira
author_facet GACHOKA, Kennedy Kiriira
author_role author
dc.contributor.advisor1.fl_str_mv BORGES, Lígia Miranda Ferreira
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/2591106734434348
dc.contributor.advisor-co1.fl_str_mv FERRI, Pedro Henrique
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/2129799749473005
dc.contributor.advisor-co2.fl_str_mv SILVA, Andrea Caetano da
dc.contributor.advisor-co2Lattes.fl_str_mv http://lattes.cnpq.br/1432918585717355
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/9231444052376453
dc.contributor.author.fl_str_mv GACHOKA, Kennedy Kiriira
contributor_str_mv BORGES, Lígia Miranda Ferreira
FERRI, Pedro Henrique
SILVA, Andrea Caetano da
dc.subject.por.fl_str_mv carrapato-estrela
comunicação química
feromônios
semioquímicos
topic carrapato-estrela
comunicação química
feromônios
semioquímicos
Cayenne tick
chemical communication
pheromones
semiochemicals
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
dc.subject.eng.fl_str_mv Cayenne tick
chemical communication
pheromones
semiochemicals
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
description Amblyomma cajennense, commonly known as Cayenne tick, is a three-host tick widely distributed in Brazil and in the rest of Latin American countries. Owing to its low host specificity, it plays an important role in the transmission of pathogens in animals and man. Chemical communication is a vital component in tick biology whereby pheromones are largely responsible in the regulation of mating, recognition of sexual partner and copulation. By understanding the behaviour of A. cajennense and the pheromones involved, there exists practical possibility for the use of these substances as an additional method of control thereby reducing the use of acaricides. Based on this, the aims of this work were to quantify and elucidate behaviours induced by pheromones emitted by females A. cajennense as well as identify and characterize qualitatively and biologically the pheromone produced by males. Engorged females were collected from naturally-infested horses for the establishment of laboratory colonies for use in this study. The pheromones were obtained by extraction in hexane and sonication by ultra-sound of fed males and females. The female sex pheromone, 2,6-dichlorophenol (2,6-DCP), was quantified by GC/MS using internal standard and standard addition methods through construction of calibration curves using 5-bromine-4-hydroxy-3-methoxybenzaldehyde and 2,6-DCP, respectively. The male extract was analysed chemically by GC/MS in comparison with standards. As concerns biological tests, glass beads were treated with two femaleequivalents of the female pheromone and 2,6-DCP in an equivalent concentration and exposed to males to observe the induced behaviour(s). Attraction to the male pheromone was evaluated by release of individual unfed, virgin females to three concentrations (1, 5 and 10 male-equivalents) of the extract in olfactometer bioassays. Orientations towards the odour source were analysed by Kruskal-Wallis and displacement angles by Circular Statistics. Aggregation was evaluated by releasing the tick stages in Petri dish bioassays shortly followed by introduction of filter paper strips impregnated with 10 male-equivalents of the extract and recording tick responses on camera. Data analysis was by Chi-square test. Tests for attachment were carried out in vivo in rabbits, Oryctolagus cuniculus, and the released ticks observed in determined intervals between 20min and 24hrs. The data was analysed by Wilcoxon signed-ranked test and Friedman s repeated measures ANOVA. The results indicated that each female emits 2.15 ng ml-1 of pheromone on average. The male extract was positive for benzoic acid, nonanoic acid, salicylic acid, benzaldehyde, 2,6-DCP, limonene and methyl salicylate. The female extract and 2,6- DCP stimulated mounting in up to 80% and 73% of all cases, respectively, confirming that 2,6-DCP alone mediates mounting behaviour in A. cajennense. There was no statistically significant attraction or aggregation in males or females induced by the male pheromone. However, significantly faster and greater attachment was recorded in both. The behavioural responses corresponded with the pheromone s composition. It was concluded that the male pheromone promotes neither attraction nor aggregation but is responsible for faster fixation of A. cajennense adults. The pheromone also contains different proportions and differs from other Amblyomma species as far as the presence or absence of certain components is concerned. The results serve as a first step in advanced studies aimed at development of novel strategies for the management of this species using these pheromones.
publishDate 2010
dc.date.available.fl_str_mv 2010-10-13
dc.date.issued.fl_str_mv 2010-08-13
dc.date.accessioned.fl_str_mv 2014-07-29T15:13:38Z
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dc.identifier.citation.fl_str_mv GACHOKA, Kennedy Kiriira. Chemical and biological pheromone of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae). 2010. 94 f. Tese (Doutorado em Ciências Agrárias) - Universidade Federal de Goiás, Goiânia, 2010.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tde/1087
identifier_str_mv GACHOKA, Kennedy Kiriira. Chemical and biological pheromone of Amblyomma cajennense (Fabricius, 1787) (Acari: Ixodidae). 2010. 94 f. Tese (Doutorado em Ciências Agrárias) - Universidade Federal de Goiás, Goiânia, 2010.
url http://repositorio.bc.ufg.br/tede/handle/tde/1087
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dc.publisher.department.fl_str_mv Ciências Agrárias
publisher.none.fl_str_mv Universidade Federal de Goiás
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