Reference 1D and 2D electrophoresis maps for potential disease related proteins in milk whey from lactating buffaloes and blood serum from buffalo calves (Water buffalo, Bubalus bubalis)

Autor(es)

Santana, Andre M. ^[1] ; Thomas, Funmilola C. ^[2] ; Silva, Daniela G. ^[1] ; McCulloch, Eilidh ^[3] ; Vidal, Ana M. C. ^[4] ; Burchmore, Richard J. S. ^[5] ; Fagliari, Jose J. ^[1] ; Eckersall, Peter D. ^[3]

Afiliação do(s) autor(es)

^[1] Sao Paulo State Univ FCAV UNESP, Sch Agr & Vet Sci, Dept Vet Clin & Surg, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP - Brazil

^[2] Fed Univ Agr, Coll Vet Med, Dept Vet Physiol & Pharmacol, Abeokuta - Nigeria

^[3] Univ Glasgow, Coll Med Vet & Life Sci, Inst Biodivers Anim Hlth & Comparat Med, Glasgow, Lanark - Scotland

^[4] Univ Sao Paulo FZEA USP, Fac Anim Sci & Food Engn, Dept Vet Med, Pirassununga, SP - Brazil

^[5] Univ Glasgow, Coll Med Vet & Life Sci, Glasgow Poly Facil, Inst Infect Immun & Inflammat, Glasgow, Lanark - Scotland

• Tipo de documento: Artigo Científico
• Fonte: Research in Veterinary Science; v. 118, p. 449-465, JUN 2018.
• Citações Web of Science: 4

Resumo

The aim of this study was to identify potential disease related proteins in milk whey of lactating buffaloes and blood serum of buffalo calves, in order to define a reference electrophoresis map for 1-DE and 2-DE. Additionally, changes in some protein patterns from buffalo calves during salmonellosis and lactating buffaloes during mastitis are presented. Milk samples were collected and distributed into groups: Milk samples from healthy buffaloes (SCC < 100.000 cells/ml, negative microbiology and CMT) (G1, n = 5) and buffaloes with subclinical mastitis (SCC > 500.000 cells/ml, positive microbiology and CMT) (G2, n = 5). Blood samples from buffalo calves (n = 6) were collected, and three calves were experimentally infected with Salmonella Dublin and samples analyzed before (MO) and 72 h after inoculation (M1). 1-DE was accomplished by loading 10 mu g of TP into SDS-PAGE, stained with Coomassie blue. 2-DE was accomplished by loading 200 mu g of TP into 11 cm, pH 3-10 non-linear IPG strips, followed by SDS-PAGE, stained with Coomassie blue. Protein bands/spots were excised, subjected to tryptic in-gel digestion and analyzed by LC/ESI-MS/MS. Protein identity was assigned using NCBI databases. After bands/spots from 1-DE and 2-DE were analyzed, a protein map with 35 and 40 different identified proteins in blood serum and milk whey, respectively, was generated. Significant changes in patterns of haptoglobin were observed in buffalo calves with salmonellosis and in patterns of IgLC, beta-lactoglobulin and alpha-lactalbumin of lactating buffaloes during mastitis. The establishment of a protein map for 1-DE and 2-DE, identifying potential disease related proteins, can help to address alterations during diseases in buffaloes. (AU)

• Processo FAPESP: 12/10094-0 - Perfil bioquímico da secreção láctea, inclusive proteínas de fase aguda e enzimas NAGase e LDH, como método auxiliar de diagnóstico e prognóstico de mastite subclínica e clínica em fêmeas bubalinas e bovinas
• Beneficiário: André Marcos Santana
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 13/26498-5 - Estudo proteômico e de proteínas de fase aguda no soro sanguíneo e soro lácteo de búfalos e bovinos: mastite e salmonelose
• Beneficiário: André Marcos Santana
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
• Processo FAPESP: 08/50388-7 - Infeccao experimental com salmonella dublin em bezerros bubalinos: estudo clinico, laboratorial e terapeutico.
• Beneficiário: André Marcos Santana
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 09/12350-0 - Estudo clínico, laboratorial e terapêutico da infecção experimental de bezerros bubalinos por Salmonella dublin
• Beneficiário: José Jurandir Fagliari
• Modalidade de apoio: Auxílio à Pesquisa - Regular