title: In vitro and in vivo evaluation of lactic acid bacteria of aquatic origin as probiotics for turbot (Scophthalmus maximus L.) farming.
abstract: Turbot (Scophthalmus maximus L.) is an important commercial marine flatfish. Its production may be affected by bacterial diseases that cause severe economical losses, mainly tenacibaculosis and vibriosis, provoked by Tenacibaculum maritimum and Vibrio splendidus, respectively. An alternative or complementary strategy to chemotherapy and vaccination for the control of these diseases is the use of probiotics. In this work, we report the in vitro and in vivo potential of eight lactic acid bacteria (LAB), previously isolated from fish, seafood and fish products intended for human consumption, as turbot probiotics. Seven out of the eight LAB exerted direct antimicrobial activity against, at least, four strains of T. maritimum and V. splendidus. All LAB survived in seawater at 18 °C for 7 days, and withstood exposure to pH 3.0 and 10% (v/v) turbot bile; however, they differed in cell surface hydrophobicity (8.2-21.7%) and in their ability to adhere to turbot skin (1.2-21.7%) and intestinal (0.7-2.1%) mucus. Most of the tested strains inhibited the binding of turbot pathogens to the mucus. Leuconostoc mesenteroides subsp. cremoris SMM69 and Weissella cibaria P71 were selected based on their strong antimicrobial activity against T. maritimum and V. splendidus, good probiotic properties, and different adhesion ability to skin mucus and capacity to inhibit the adhesion of turbot pathogens to mucus. These two LAB strains were harmless when administered by bath to turbot larvae and juveniles; moreover, real-time PCR on the transcription levels of the immunity-related genes encoding IL-1β, TNF-α, lysozyme, C3, MHC-Iα and MHC-IIα in five organs (head-kidney, spleen, liver, intestine and skin) revealed the ability of these LAB to stimulate their expression in turbot juveniles, especially the non-specific immunity associated genes in mucosal tissues. Based on our results, Lc. cremoris SMM69 and W. cibaria P71 may be considered as suitable probiotic candidates for turbot farming.
authors.affiliation: Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain.
authors.forename: Estefanía
authors.initials: E
authors.lastname: Muñoz-Atienza
authors.affiliation: Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain; Institute for Biotechnology and Bioengineering, Centre of Genomics and Biotechnology, University of Trás-os-Montes and Alto Douro, 5001-801 Vila Real, Portugal.
authors.forename: Carlos
authors.initials: C
authors.lastname: Araújo
authors.affiliation: Centro Oceanográfico de Vigo, Instituto Español de Oceanografía (IEO), 36390 Vigo, Pontevedra, Spain.
authors.forename: Susana
authors.initials: S
authors.lastname: Magadán
authors.affiliation: Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain.
authors.forename: Pablo E
authors.initials: PE
authors.lastname: Hernández
authors.affiliation: Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Herranz
authors.affiliation: Department of Microbiology and Parasitology, Faculty of Biology, University of Santiago de Compostela, 15782-Santiago de Compostela, Spain.
authors.forename: Ysabel
authors.initials: Y
authors.lastname: Santos
authors.affiliation: Grupo de Seguridad y Calidad de los Alimentos por Bacterias Lácticas, Bacteriocinas y Probióticos (Grupo SEGABALBP), Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040-Madrid, Spain. Electronic address: lcintas@vet.ucm.es.
authors.forename: Luis M
authors.initials: LM
authors.lastname: Cintas
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D017756:Aquaculture / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D005393:Fish Diseases / Q000382:microbiology* / Q000517:prevention & control*; D005412:Flatfishes / Q000254:growth & development*; D045826:Flavobacteriaceae Infections / Q000517:prevention & control / Q000662:veterinary*; D066298:In Vitro Techniques; D056546:Lactobacillales / Q000235:genetics; D019936:Probiotics / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D060493:Tenacibaculum / Q000187:drug effects; D014733:Vibrio / Q000187:drug effects; D014735:Vibrio Infections / Q000517:prevention & control / Q000662:veterinary*
chemical_list: D017931:DNA Primers
keywords: Functional properties; Immunity-related gene expression; Lactic acid bacteria; Probiotics; Turbot (Scophthalmus maximus L.)