Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/17968
Title: Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus
Authors: Golo, Patrícia Silva
Santos, Alessa Siqueira de Oliveira dos
Monteiro, Caio Marcio Oliveira
Perinotto, Wendell Marcelo de Souza
Quinelato, Simone
Camargo, Mariana Guedes
Sá, Fillipe Araujo de
Angelo, Isabele da Costa
Martins, Marta Fonseca
Prata, Marcia Cristina de Azevedo
Bittencourt, Vânia Rita Elias Pinheiro
Affilliation: Universidade Federal Rural do Rio de Janeiro. Programa de Pós-graduação em Ciências Veterinárias. Seropédica, RJ, Brasil.
Embrapa Gado de Leite. Juiz de Fora, MG, Brasil.
Universidade Federal Rural do Rio de Janeiro. Programa de Pós-graduação em Ciências Veterinárias. Seropédica, RJ, Brasil / Universidade Federal de Juiz de Fora. Campus Universitário Bairro Martelos. Juiz de Fora, MG, Brasil / Universidade Federal de Goiás. Programa de Pós-graduação em Ciência Animal. Goiânia, GO, Brasil.
Universidade de Cuiabá. Faculdade de Medicina Veterinária. Mestrado em Biociência Animal. Cuiabá, MT, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Coleção de Culturas de Fungos Filamentosos. Rio de Janeiro, RJ. Brasil.
Universidade Federal Rural do Rio de Janeiro. Programa de Pós-graduação em Ciências Veterinárias. Seropédica, RJ, Brasil.
iversidade Federal do Rio de Janeiro. Instituto de Bioquímica Médica Leopoldo de Meis. Curso de Pós-Graduação em Química Biológicas. Rio de Janeiro, RJ, Brasil.
Universidade Federal Rural do Rio de Janeiro. Programa de Pós-graduação em Ciências Veterinárias. Seropédica, RJ, Brasil.
Embrapa Gado de Leite. Juiz de Fora, MG, Brasil.
Embrapa Gado de Leite. Juiz de Fora, MG, Brasil.
niversidade Federal Rural do Rio de Janeiro. Programa de Pós-graduação em Ciências Veterinárias. Seropédica, RJ, Brasil.
Abstract: In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.
Keywords: Gel electrophoresis
Biological control
Cattle tick
Metarhizium
Heterorhabditis
Protein bands
keywords: Eletroforese em gel
Controle biológico
Carrapatos
Doenças dos Bovinos
Metarhizium
Bandas de proteínas
Issue Date: 2016
Publisher: Springer Verlag (Germany)
Citation: GOLO, Patrícia Silva; et al. Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus. Parasitol Res, v. 115, n.9, p.3459–3468, Sept. 2016.
DOI: 10.1007/s00436-016-5109-z
ISSN: 0932-0113
Copyright: restricted access
Appears in Collections:IOC - Artigos de Periódicos

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