Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B

Detalhes bibliográficos
Autor(a) principal: Bernardo, Cíntia das Chagas
Data de Publicação: 2016
Tipo de documento: Tese
Idioma: por
Título da fonte: Biblioteca Digital de Teses e Dissertações da UFG
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/5993
Resumo: The current study compared the virulence of conidia and blastospores of Metarhizium anisopliae s.l. (IP 363), Metarhizium robertsii (IP 146) and Beauveria bassiana s.l. (IP 361 and GC 307) against Rhipicephalus microplus, and the tolerance to heat and UV-B radiation; in addition, it evaluated the development of conidia and blastospores of the isolates IP 146 and IP 361 on the tick cuticle by scanning electron microscopy (SEM). Larvae and engorged females of R. microplus were treated (larvae: 106, 107 or 108 propagules/mL, females: 107 propagules/mL) by immersion in conidia or blastospores suspensions of tested isolates. The higher percentage of larval mortality was obtained in the group treated with conidia of IP 361, which had lower LC50. In bioassays with engorged females, IP 146 and IP 361 blastospores, provided tick percent control superior to 90%, while conidia of the same isolates, promoted 70.97% and 63.29% of tick control, respectively. Ticks were treated topically with 50 μL of fungal suspension (107 propagules/mL) and incubated at different times from 0 to 96 hours to analyze its development on the engorged females cuticle; after each incubation time, the females were fixed and analyzed by SEM. Blastospores of both isolates have started development 4 hours after treatment, demonstrating rapid development and suggesting penetration by the tick natural openings; at 4 hours incubation, indicative of penetration of IP 361 blastospores through the cuticle was observed, but no signs of penetration was observed with IP 146 blastospores at any time evaluated. Furthermore, fungal suspensions (103 propagules/mL) were exposed to heat (45 °C) for several time periods, then inoculated in Petri plates with BDAY medium plus chloramphenicol (0.055% v/v), and incubated for 7 days at 27 °C and RH ≥ 80%. ARSEF 324 conidia (79.1%) were more tolerant to heat than conidia of IP 363 (55.5%), IP 146 (1.5%), GC 307 (0%), and IP 361 (0% ) at 2 hours exposure, as well as blastospores after 60 minutes exposure, demonstrating mean percent CFU of 100%, 12.3%, 30.7%, 55% and 0%, respectively. Fungal suspensions (103 propagules/mL) were also inoculated on BDAY in Petri plates, and exposed to UV-B radiation; after treatment, plates were incubated for 7 days at 27 °C and RH ≥ 80%. No difference in mean relative percent CFU between conidia and blastospores was observed. Suitable adjuvants which aim at protecting fungal propagules against stressful abiotic factors are required for conidia and blastospores; however, the selection of isolates with marked natural tolerance to heat and UV-B radiation may increase performance of bioproducts. Accordingly, it is suggested that blastospores are promising fungal propagules for biological control of ticks, since they were virulent against R. microplus; in addition, the rapid development of blastospores on the tick cuticle indicates they may be exposed shortly to harmful environmental abiotic factors.
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spelling Fernandes, Éverton Kort Kamphttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4707291J3Fernandes, Éverton Kort KampRangel, Drauzio Eduardo NarettoMascarin, Gabriel MouraBittencourt, Vânia Rita Elias PinheiroArruda, Walquíriahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4248596Y6Bernardo, Cíntia das Chagas2016-08-25T12:47:45Z2016-07-19BERNARDO, C. C. Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B. 2016. 93 f. Tese (Doutorado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2016.http://repositorio.bc.ufg.br/tede/handle/tede/5993The current study compared the virulence of conidia and blastospores of Metarhizium anisopliae s.l. (IP 363), Metarhizium robertsii (IP 146) and Beauveria bassiana s.l. (IP 361 and GC 307) against Rhipicephalus microplus, and the tolerance to heat and UV-B radiation; in addition, it evaluated the development of conidia and blastospores of the isolates IP 146 and IP 361 on the tick cuticle by scanning electron microscopy (SEM). Larvae and engorged females of R. microplus were treated (larvae: 106, 107 or 108 propagules/mL, females: 107 propagules/mL) by immersion in conidia or blastospores suspensions of tested isolates. The higher percentage of larval mortality was obtained in the group treated with conidia of IP 361, which had lower LC50. In bioassays with engorged females, IP 146 and IP 361 blastospores, provided tick percent control superior to 90%, while conidia of the same isolates, promoted 70.97% and 63.29% of tick control, respectively. Ticks were treated topically with 50 μL of fungal suspension (107 propagules/mL) and incubated at different times from 0 to 96 hours to analyze its development on the engorged females cuticle; after each incubation time, the females were fixed and analyzed by SEM. Blastospores of both isolates have started development 4 hours after treatment, demonstrating rapid development and suggesting penetration by the tick natural openings; at 4 hours incubation, indicative of penetration of IP 361 blastospores through the cuticle was observed, but no signs of penetration was observed with IP 146 blastospores at any time evaluated. Furthermore, fungal suspensions (103 propagules/mL) were exposed to heat (45 °C) for several time periods, then inoculated in Petri plates with BDAY medium plus chloramphenicol (0.055% v/v), and incubated for 7 days at 27 °C and RH ≥ 80%. ARSEF 324 conidia (79.1%) were more tolerant to heat than conidia of IP 363 (55.5%), IP 146 (1.5%), GC 307 (0%), and IP 361 (0% ) at 2 hours exposure, as well as blastospores after 60 minutes exposure, demonstrating mean percent CFU of 100%, 12.3%, 30.7%, 55% and 0%, respectively. Fungal suspensions (103 propagules/mL) were also inoculated on BDAY in Petri plates, and exposed to UV-B radiation; after treatment, plates were incubated for 7 days at 27 °C and RH ≥ 80%. No difference in mean relative percent CFU between conidia and blastospores was observed. Suitable adjuvants which aim at protecting fungal propagules against stressful abiotic factors are required for conidia and blastospores; however, the selection of isolates with marked natural tolerance to heat and UV-B radiation may increase performance of bioproducts. Accordingly, it is suggested that blastospores are promising fungal propagules for biological control of ticks, since they were virulent against R. microplus; in addition, the rapid development of blastospores on the tick cuticle indicates they may be exposed shortly to harmful environmental abiotic factors.O presente estudo comparou conídios e blastosporos de Metarhizium anisopliae s.l. (IP 363), Metarhizium robertsii (IP 146) e Beauveria bassiana s.l. (IP 361 e CG 307) quanto a virulência para Rhipicephalus microplus, e quanto a tolerância ao calor e a radiação UV-B; foi avaliado ainda o desenvolvimento de conídios e blastosporos dos isolados IP 146 e IP 361 na cutícula de carrapatos por meio da microscopia eletrônica de varredura (MEV). Larvas e fêmeas ingurgitadas de R. microplus foram tratadas (larvas: 106, 107 ou 108 propágulos/mL; fêmeas: 107 propágulos/mL) por imersão, com conídios e blastosporos dos isolados avaliados. Maior percentual de mortalidade foi obtido nos grupos de larvas tratadas com conídios de IP 361, o qual obteve menor CL50. No bioensaio com fêmeas, blastosporos de IP 146 e IP 361 proporcionaram percentual de controle superior a 90%, enquanto que conídios dos mesmos isolados promoveram mortalidade de 70,97% e 63,29%, respectivamente. Para análise do desenvolvimento dos propágulos na cutícula de fêmeas ingurgitadas, essas foram tratadas topicamente com 50 µL de suspensão fúngica (107 propágulos/mL), incubadas em diferentes tempos de 0 a 96 horas; após cada tempo de incubação as fêmeas foram processadas e analisadas por MEV. Blastosporos de ambos isolados iniciaram seu desenvolvimento já após 4 horas de tratamento, demonstrando rápido desenvolvimento e sugerindo penetração por aberturas naturais do carrapato; com 4 horas de incubação foi possível ver a penetração de blastosporos de IP 361 através da cutícula, situação não evidenciada pelo isolado IP 146 em nenhum tempo avaliado. Nos testes de tolerância ao calor, o isolado de Metarhizium acridum (ARSEF 324) foi inserido como isolado padrão, por ser conhecida a sua tolerância. Suspensões fúngicas (103 propágulos/mL) foram expostas ao calor (45°C) por diferentes tempos, em seguida inoculadas em placas de Petri com meio BDAY acrescido de cloranfenicol (0,055% v/v) e incubadas por 7 dias a 27°C, e UR ≥ 80%. Conídios de ARSEF 324 (79,1%) demonstraram ser mais tolerantes ao calor do que conídios de IP 363 (55,5%), IP 146 (1,5%), CG 307 (0%) e IP 361 (0%) no tempo de 2 horas de exposição, assim como blastosporos no tempo de 60 minutos, demonstrando percentual relativo médio de 100%, 12,3%, 30,7%, 55% e 0%, respectivamente. Nos testes de exposição à UV-B, suspensões fúngicas (103 propágulos/mL) foram inoculadas em placas de Petri contendo BDAY e expostas a diferentes doses de radiação; após tratamento, as placas foram incubadas por 7 dias a 27°C, e UR ≥ 80%. Não houve diferença do percentual relativo médio de conídios e blastosporos expostos a mesma dose de radiação. Adjuvantes adequados que visem proteger os propágulos fúngicos contra fatores abióticos estressantes são requeridos para formulações de conídios e blastosporos; no entanto, a seleção de isolados naturalmente mais tolerantes ao calor e a radiação UV-B podem favorecer o desenvolvimento de bioprodutos. Nesse sentido, acredita-se que blastosporos sejam promissores para o biocontrole de carrapatos, já que estes se demonstraram virulentos para R. microplus, além de apresentarem rápido desenvolvimento sobre a cutícula desse artrópode, o que pode indicar menor tempo de exposição desses propágulos a fatores abióticos limitantes no ambiente.Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-25T12:47:20Z No. of bitstreams: 2 Tese - Cíntia das Chagas Bernardo - 2016.pdf: 2168975 bytes, checksum: f20144021bf112df62693342cfba7dae (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-08-25T12:47:45Z (GMT) No. of bitstreams: 2 Tese - Cíntia das Chagas Bernardo - 2016.pdf: 2168975 bytes, checksum: f20144021bf112df62693342cfba7dae (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2016-08-25T12:47:45Z (GMT). No. of bitstreams: 2 Tese - Cíntia das Chagas Bernardo - 2016.pdf: 2168975 bytes, checksum: f20144021bf112df62693342cfba7dae (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-07-19Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESapplication/pdfporUniversidade Federal de GoiásPrograma de Pós-graduação em Medicina Tropical e Saúde Publica (IPTSP)UFGBrasilInstituto de Patologia Tropical e Saúde Pública - IPTSP (RG)http://creativecommons.org/licenses/by-nc-nd/4.0/info:eu-repo/semantics/openAccessPropágulos fúngicosFungo entomopatogênicoIxodidaeTermotolerânciaRadiação ultravioletaCarrapatoFungal propagulesEntomopathogenic fungiTickIxodidaeThermotoleranceUV radiation.CIENCIAS BIOLOGICAS::PARASITOLOGIAConídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-BConidia and blastospores of Metarhizium spp. and Beauveria bassiana: virulence for Rhipicephalus microplus and response to heat and UV-B radiationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis6085308344741430434600600600600-7769011444564556288-45445767472715743062075167498588264571reponame:Biblioteca Digital de Teses e Dissertações da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
dc.title.alternative.eng.fl_str_mv Conidia and blastospores of Metarhizium spp. and Beauveria bassiana: virulence for Rhipicephalus microplus and response to heat and UV-B radiation
title Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
spellingShingle Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
Bernardo, Cíntia das Chagas
Propágulos fúngicos
Fungo entomopatogênico
Ixodidae
Termotolerância
Radiação ultravioleta
Carrapato
Fungal propagules
Entomopathogenic fungi
Tick
Ixodidae
Thermotolerance
UV radiation.
CIENCIAS BIOLOGICAS::PARASITOLOGIA
title_short Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
title_full Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
title_fullStr Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
title_full_unstemmed Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
title_sort Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B
author Bernardo, Cíntia das Chagas
author_facet Bernardo, Cíntia das Chagas
author_role author
dc.contributor.advisor1.fl_str_mv Fernandes, Éverton Kort Kamp
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4707291J3
dc.contributor.referee1.fl_str_mv Fernandes, Éverton Kort Kamp
dc.contributor.referee2.fl_str_mv Rangel, Drauzio Eduardo Naretto
dc.contributor.referee3.fl_str_mv Mascarin, Gabriel Moura
dc.contributor.referee4.fl_str_mv Bittencourt, Vânia Rita Elias Pinheiro
dc.contributor.referee5.fl_str_mv Arruda, Walquíria
dc.contributor.authorLattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4248596Y6
dc.contributor.author.fl_str_mv Bernardo, Cíntia das Chagas
contributor_str_mv Fernandes, Éverton Kort Kamp
Fernandes, Éverton Kort Kamp
Rangel, Drauzio Eduardo Naretto
Mascarin, Gabriel Moura
Bittencourt, Vânia Rita Elias Pinheiro
Arruda, Walquíria
dc.subject.por.fl_str_mv Propágulos fúngicos
Fungo entomopatogênico
Ixodidae
Termotolerância
Radiação ultravioleta
Carrapato
topic Propágulos fúngicos
Fungo entomopatogênico
Ixodidae
Termotolerância
Radiação ultravioleta
Carrapato
Fungal propagules
Entomopathogenic fungi
Tick
Ixodidae
Thermotolerance
UV radiation.
CIENCIAS BIOLOGICAS::PARASITOLOGIA
dc.subject.eng.fl_str_mv Fungal propagules
Entomopathogenic fungi
Tick
Ixodidae
Thermotolerance
UV radiation.
dc.subject.cnpq.fl_str_mv CIENCIAS BIOLOGICAS::PARASITOLOGIA
description The current study compared the virulence of conidia and blastospores of Metarhizium anisopliae s.l. (IP 363), Metarhizium robertsii (IP 146) and Beauveria bassiana s.l. (IP 361 and GC 307) against Rhipicephalus microplus, and the tolerance to heat and UV-B radiation; in addition, it evaluated the development of conidia and blastospores of the isolates IP 146 and IP 361 on the tick cuticle by scanning electron microscopy (SEM). Larvae and engorged females of R. microplus were treated (larvae: 106, 107 or 108 propagules/mL, females: 107 propagules/mL) by immersion in conidia or blastospores suspensions of tested isolates. The higher percentage of larval mortality was obtained in the group treated with conidia of IP 361, which had lower LC50. In bioassays with engorged females, IP 146 and IP 361 blastospores, provided tick percent control superior to 90%, while conidia of the same isolates, promoted 70.97% and 63.29% of tick control, respectively. Ticks were treated topically with 50 μL of fungal suspension (107 propagules/mL) and incubated at different times from 0 to 96 hours to analyze its development on the engorged females cuticle; after each incubation time, the females were fixed and analyzed by SEM. Blastospores of both isolates have started development 4 hours after treatment, demonstrating rapid development and suggesting penetration by the tick natural openings; at 4 hours incubation, indicative of penetration of IP 361 blastospores through the cuticle was observed, but no signs of penetration was observed with IP 146 blastospores at any time evaluated. Furthermore, fungal suspensions (103 propagules/mL) were exposed to heat (45 °C) for several time periods, then inoculated in Petri plates with BDAY medium plus chloramphenicol (0.055% v/v), and incubated for 7 days at 27 °C and RH ≥ 80%. ARSEF 324 conidia (79.1%) were more tolerant to heat than conidia of IP 363 (55.5%), IP 146 (1.5%), GC 307 (0%), and IP 361 (0% ) at 2 hours exposure, as well as blastospores after 60 minutes exposure, demonstrating mean percent CFU of 100%, 12.3%, 30.7%, 55% and 0%, respectively. Fungal suspensions (103 propagules/mL) were also inoculated on BDAY in Petri plates, and exposed to UV-B radiation; after treatment, plates were incubated for 7 days at 27 °C and RH ≥ 80%. No difference in mean relative percent CFU between conidia and blastospores was observed. Suitable adjuvants which aim at protecting fungal propagules against stressful abiotic factors are required for conidia and blastospores; however, the selection of isolates with marked natural tolerance to heat and UV-B radiation may increase performance of bioproducts. Accordingly, it is suggested that blastospores are promising fungal propagules for biological control of ticks, since they were virulent against R. microplus; in addition, the rapid development of blastospores on the tick cuticle indicates they may be exposed shortly to harmful environmental abiotic factors.
publishDate 2016
dc.date.accessioned.fl_str_mv 2016-08-25T12:47:45Z
dc.date.issued.fl_str_mv 2016-07-19
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
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dc.identifier.citation.fl_str_mv BERNARDO, C. C. Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B. 2016. 93 f. Tese (Doutorado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2016.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/5993
identifier_str_mv BERNARDO, C. C. Conídios e blastosporos de Metarhizium spp. e Beauveria bassiana: virulência para Rhipicephalus microplus e resposta ao calor e à radiação UV-B. 2016. 93 f. Tese (Doutorado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2016.
url http://repositorio.bc.ufg.br/tede/handle/tede/5993
dc.language.iso.fl_str_mv por
language por
dc.relation.program.fl_str_mv 6085308344741430434
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dc.relation.sponsorship.fl_str_mv 2075167498588264571
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
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rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
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dc.publisher.none.fl_str_mv Universidade Federal de Goiás
dc.publisher.program.fl_str_mv Programa de Pós-graduação em Medicina Tropical e Saúde Publica (IPTSP)
dc.publisher.initials.fl_str_mv UFG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Instituto de Patologia Tropical e Saúde Pública - IPTSP (RG)
publisher.none.fl_str_mv Universidade Federal de Goiás
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