title: Nitric oxide synthase-dependent immune response against gram negative bacteria in a crustacean, Litopenaeus vannamei.
abstract: Nitric oxide (NO) is a short-lived radical generated by nitric oxide synthases (NOS). NO is involved in a variety of functions in invertebrates, including host defense. In previous studies, we isolated and sequenced for the first time the NOS gene from hemocytes of Panulirus argus, demonstrating the inducibility of this enzyme by lipopolysaccharide in vitro e in vivo. Hyperimmune serum was obtained from rabbits immunized with a P. argus -NOS fragment of 31 kDa produced in Escherichia coli, which specifically detected the recombinant polypeptide and the endogenous NOS from lobster hemocytes by western blotting and immunofluorescence. In the present work, we demonstrate that the hyperimmune serum obtained against P. argus NOS also recognizes Litopenaeus vannamei NOS in hemocytes by western blotting and immunofluorescence. Our data also show that while the hemolymph of L. vannamei has a strong antibacterial activity against the Gram negative bacteria Aeromonas hydrophila, the administration of the anti NOS serum reduce the natural bacterial clearance. These results strongly suggest that NOS is required for the shrimp immune defense toward Gram negative bacteria. Therefore, the monitoring of induction of NOS could be an important tool for testing immunity in shrimp farming.
authors.affiliation: Center for Marine Research, University of Havana (CIM-UH), Cuba.
authors.forename: Tania
authors.initials: T
authors.lastname: Rodríguez-Ramos
authors.affiliation: Center for Genetic Engeneering and Biotechnology (CIGB), Cuba.
authors.forename: Yamila
authors.initials: Y
authors.lastname: Carpio
authors.affiliation: Departamento de Biomedicina, Biotecnología y Salud Pública, Faculty of Sciences, University of Cadiz (UCA), Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Bolívar
authors.affiliation: Center for Genetic Engeneering and Biotechnology (CIGB), Cuba.
authors.forename: Leonardo
authors.initials: L
authors.lastname: Gómez
authors.affiliation: Center for Genetic Engeneering and Biotechnology (CIGB), Cuba. Electronic address: mario.pablo@cigb.edu.cu.
authors.forename: Mario Pablo
authors.initials: MP
authors.lastname: Estrada
authors.affiliation: Departamento de Biomedicina, Biotecnología y Salud Pública, Faculty of Sciences, University of Cadiz (UCA), Spain. Electronic address: carlos.pendon@uca.es.
authors.forename: Carlos
authors.initials: C
authors.lastname: Pendón
mesh_terms: D016980:Aeromonas hydrophila / Q000502:physiology*; D000818:Animals; D000890:Anti-Infective Agents / Q000378:metabolism; D060829:Arthropod Proteins / Q000378:metabolism*; D006458:Hemolymph / Q000276:immunology; D007113:Immunity, Innate; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D033561:Penaeidae / Q000235:genetics* / Q000276:immunology* / Q000382:microbiology
chemical_list: D000890:Anti-Infective Agents; D060829:Arthropod Proteins; D019001:Nitric Oxide Synthase
keywords: Aeromonas hydrophila; Invertebrate immunity; Litopenaeus vannamei; NOS; Nitric oxide synthase