title: Antibacterial activity of thionin Thi2.1 from Arabidopsis thaliana expressed by bovine endothelial cells against Staphylococcus aureus isolates from bovine mastitis.
abstract: Bovine mastitis is mainly caused by Staphylococcus aureus and antimicrobial therapy, commonly used for its control, has resulted in an increase in the frequency of resistant staphylococci in recent years. Thus, alternative therapies are desirable and the antimicrobial peptides represent attractive control agents. In this work, we expressed the antimicrobial peptide thionin Thi2.1 cDNA from Arabidopsis thaliana in the bovine endothelial cell line BVE-E6E7 and evaluated its activity against bovine mastitis S. aureus isolates. A polyclonal population from BVE-E6E7 cells transfected with the pThi2.1 construct was obtained and thionin Thi2.1 expression was confirmed by RT-PCR. From this population, eight stably transfected cell clones were obtained and their conditioned media (CM) were evaluated against the S. aureus ATCC 27543 strain. Clones showed high antibacterial activity (>95%) relative to the activity of the polyclonal population. The C8 clone showed the highest antibacterial activity (>99%) and its CM was evaluated against eleven bovine mastitis S. aureus isolates. A 2.5microg aliquot of total protein from the C8 clone's CM inhibited the growth of S. aureus isolates (>40%) relative to the CM from BVE-E6E7 cells used as control. Growth inhibition of S. aureus isolates was dose-dependent, showing a total inhibition at concentrations higher than 3.12microg/ml. These results suggest that thionin Thi2.1 antimicrobial peptide could be use in the treatment of bovine mastitis.
authors.affiliation: Centro Multidisciplinario de Estudios en Biotecnología, Facultad de Medicina Veterinaria y Zootecnia, Universidad Michoacana de San Nicolás de Hidalgo, Apdo. Postal 53, Administración Chapultepec, C.P. 58262 Morelia, Michoacán, México.
authors.forename: Alejandra
authors.initials: A
authors.lastname: Ochoa-Zarzosa
authors.forename: Heber
authors.initials: H
authors.lastname: Loeza-Angeles
authors.forename: Eduardo
authors.initials: E
authors.lastname: Sagrero-Cisneros
authors.forename: Erik
authors.initials: E
authors.lastname: Villagómez-Gómez
authors.forename: Leticia
authors.initials: L
authors.lastname: Lara-Zárate
authors.forename: Joel E
authors.initials: JE
authors.lastname: López-Meza
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology* / Q000627:therapeutic use; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D017360:Arabidopsis / Q000737:chemistry*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000494:pharmacology* / Q000627:therapeutic use; D002417:Cattle; D004305:Dose-Response Relationship, Drug; D024881:Drug Resistance, Bacterial; D042783:Endothelial Cells; D005260:Female; D008414:Mastitis, Bovine / Q000188:drug therapy / Q000382:microbiology*; D008826:Microbial Sensitivity Tests / Q000662:veterinary; D013203:Staphylococcal Infections / Q000188:drug therapy / Q000382:microbiology / Q000662:veterinary*; D013211:Staphylococcus aureus / Q000187:drug effects*; D014162:Transfection
chemical_list: D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D029681:Arabidopsis Proteins; C097482:THI2.1 protein, Arabidopsis