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Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus

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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.

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Acknowledgments

This study was supported by grants and fellowships from: Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Programa de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) from Rio de Janeiro State, Brazil. Vânia R.E.P. Bittencourt is a CNPq researcher.

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Authors and Affiliations

  1. Programa de Pós-graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, 23890-000, Seropédica, RJ, Brazil

    Patrícia Silva Golo, Caio Marcio Oliveira Monteiro, Mariana Guedes Camargo, Isabele da Costa Angelo & Vânia Rita Elias Pinheiro Bittencourt

  2. Embrapa Gado de Leite, Rua Eugênio do Nascimento, 610 Bairro Dom Bosco, 36038-330, Juiz de Fora, Minas Gerais, Brazil

    Alessa Siqueira de Oliveira dos Santos, Marta Fonseca Martins & Marcia Cristina de Azevedo Prata

  3. Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, s/n - Campus Universitário Bairro Martelos, 36036-330, Juiz de Fora, MG, Brazil

    Caio Marcio Oliveira Monteiro

  4. Programa de Pós-graduação em Cência Animal da Universidade Federal de Goiás, Avenida Esperança, s/n, Campus Samambaia, 74690-900, Goiânia, GO, Brazil

    Caio Marcio Oliveira Monteiro

  5. Faculdade de Medicina Veterinária, Mestrado em Biociência Animal, Universidade de Cuiabá, Avenida Beira Rio, 3100 Jardim Europa, 78015-480, Cuiabá, MT, Brazil

    Wendell Marcelo de Souza Perinotto

  6. Fundação Osvaldo Cruz, Coleção de Culturas de Fungos Filamentosos, Pavilhão Rocha Lima - sala 525. Avenida Brasil, 4365 Manguinhos, 21040-900, Rio de Janeiro, RJ, Brazil

    Simone Quinelato

  7. Curso de Pós-Graduação em Química Biológica, Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro, Avenida Carlos Chagas Filho, 373, 21941-902, Rio de Janeiro, RJ, Brazil

    Fillipe Araujo de Sá

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  1. Patrícia Silva Golo
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  2. Alessa Siqueira de Oliveira dos Santos
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  3. Caio Marcio Oliveira Monteiro
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  8. Isabele da Costa Angelo
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Correspondence to Patrícia Silva Golo.

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Golo, P.S., dos Santos, A.S.d.O., Monteiro, C.M.O. 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 115, 3459–3468 (2016). https://doi.org/10.1007/s00436-016-5109-z

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