NDLI: Identification of Vibrio harveyi proteins involved in the specific immune response of Senegalese sole (Solea senegalensis, Kaup).

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Identification of Vibrio harveyi proteins involved in the specific immune response of Senegalese sole (Solea senegalensis, Kaup).

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Medina, A. Mancera, J. M. Martínez-Manzanares, E. Moriñigo, M. A. Arijo, S.
Description	Country affiliation: Spain Author Affiliation: Medina A ( Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, Spain.); Mancera JM ( Departamento de Biología, Facultad de Ciencias del Mar y Ambientales, Universidad de Cádiz, Campus de Excelencia Internacional del Mar, 11510 Puerto Real, Cádiz, Spain.); Martínez-Manzanares E ( Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, Spain.); Moriñigo MA ( Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, Spain.); Arijo S ( Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 Málaga, Spain. Electronic address: sarijo@uma.es.)
Abstract	Senegalese sole cultures are frequently affected by Vibrio harveyi disease outbreaks. Vaccines in aquaculture are one of the most successful methods of preventing fish pathologies; however, these vaccines are usually composed of inactivated whole cells containing a wide pool of antigens, and some do not induce any protection against pathogens. Thus, the aim of this study was to identify immunogenic proteins of V. harveyi involved in the specific antibody production by Senegalese sole. S. senegalensis specimens were immunized, by intraperitoneal injection, with V. harveyi bacterin supplemented with inactivated extracellular polymeric substances (ECP) and Freund incomplete adjuvant to obtain polyclonal antiserum. One month later, specimens were re-inoculated with the same antigens. Sera from immunized fish were collected two months post first immunization. Strong specific immune response to V. harveyi antigens was detected by ELISA using bacterin (limit dilutions of sera were 1:64000), ECP (1:4000) and outer membrane proteins (OMP) (1:4000) as antigens. Presence of immunogenic proteins in V. harveyi ECP and OMP were determined by 2D-PAGE. For Western Blot analysis some gels were transferred onto nitrocellulose membranes and incubated with sera from S. senegalensis specimens immunized against V. harveyi. 2D-PAGE and Western Blot showed at least five reactive proteins in the ECP and two in the OMP fraction. The spots that clearly reacted with the sole antiserum were excised from stained gel, and analyzed by mass spectrometry (MALDI/TOFTOF). A database search was then performed, using MASCOT as the search method. According to the results, the five ECP spots were identified as Maltoporine, protein homologous to Metal dependent phosphohydrolase, two porins isoforms of V. harveyi and a protein homologous to the cell division protein FtsH. Reactive proteins in the OMP fraction were identified as the protein 3-hydroxyisobutyrate dehydrogenase and a protein homologous to acid phosphatase.
File Format	HTM / HTML
ISSN	10504648
Issue Number	1
Volume Number	47
e-ISSN	10959947
Journal	Fish & Shellfish Immunology
Language	English
Publisher	Elsevier
Publisher Date	2015-11-01
Publisher Place	Great Britain (UK)
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Marine Biology Discipline Immunology Bacterial Proteins Genetics Fish Diseases Immunology Flatfishes Immunity, Innate Vibrio Infections Veterinary Vibrio Physiology Animals Antibodies, Bacterial Blood Metabolism Bacterial Vaccines Pharmacology Microbiology Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Environmental Chemistry Medicine Aquatic Science Immunology

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