title: Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus.
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.
authors.affiliation: Programa de Pós-graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, 23890-000, Seropédica, RJ, Brazil. patriciagolo@gmail.com.
authors.forename: Patrícia Silva
authors.initials: PS
authors.lastname: Golo
authors.affiliation: Embrapa Gado de Leite, Rua Eugênio do Nascimento, 610 Bairro Dom Bosco, 36038-330, Juiz de Fora, Minas Gerais, Brazil.
authors.forename: Alessa Siqueira de Oliveira
authors.initials: AS
authors.lastname: Dos Santos
authors.affiliation: Programa de Pós-graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, 23890-000, Seropédica, RJ, Brazil.|Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, s/n - Campus Universitário Bairro Martelos, 36036-330, Juiz de Fora, MG, Brazil.|Programa de Pós-graduação em Cência Animal da Universidade Federal de Goiás, Avenida Esperança, s/n, Campus Samambaia, 74690-900, Goiânia, GO, Brazil.
authors.forename: Caio Marcio Oliveira
authors.initials: CM
authors.lastname: Monteiro
authors.affiliation: Faculdade de Medicina Veterinária, Mestrado em Biociência Animal, Universidade de Cuiabá, Avenida Beira Rio, 3100 Jardim Europa, 78015-480, Cuiabá, MT, Brazil.
authors.forename: Wendell Marcelo de Souza
authors.initials: WM
authors.lastname: Perinotto
authors.affiliation: 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.
authors.forename: Simone
authors.initials: S
authors.lastname: Quinelato
authors.affiliation: Programa de Pós-graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, 23890-000, Seropédica, RJ, Brazil.
authors.forename: Mariana Guedes
authors.initials: MG
authors.lastname: Camargo
authors.affiliation: 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.
authors.forename: Fillipe Araujo
authors.initials: FA
authors.lastname: de Sá
authors.affiliation: Programa de Pós-graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, 23890-000, Seropédica, RJ, Brazil.
authors.forename: Isabele da Costa
authors.initials: Ida C
authors.lastname: Angelo
authors.affiliation: Embrapa Gado de Leite, Rua Eugênio do Nascimento, 610 Bairro Dom Bosco, 36038-330, Juiz de Fora, Minas Gerais, Brazil.
authors.forename: Marta Fonseca
authors.initials: MF
authors.lastname: Martins
authors.affiliation: Embrapa Gado de Leite, Rua Eugênio do Nascimento, 610 Bairro Dom Bosco, 36038-330, Juiz de Fora, Minas Gerais, Brazil.
authors.forename: Marcia Cristina de Azevedo
authors.initials: MC
authors.lastname: Prata
authors.affiliation: Programa de Pós-graduação em Ciências Veterinárias, Universidade Federal Rural do Rio de Janeiro, Rodovia BR 465, 23890-000, Seropédica, RJ, Brazil.
authors.forename: Vânia Rita Elias Pinheiro
authors.initials: VR
authors.lastname: Bittencourt
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods*; D005260:Female; D005658:Fungi / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006458:Hemolymph / Q000737:chemistry / Q000378:metabolism; D056656:Lab-On-A-Chip Devices; D009348:Nematoda / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology; D015203:Reproducibility of Results; D048494:Rhipicephalus / Q000737:chemistry* / Q000378:metabolism / Q000382:microbiology* / Q000469:parasitology*
chemical_list: D060829:Arthropod Proteins
keywords: Biological control; Cattle tick; Gel electrophoresis; Heterorhabditis; Metarhizium; Protein bands