ABSTRACT
BACKGROUND: Vibriosis is one of the most ubiquitous fish diseases caused by bacteria belonging to the genus Vibrio such as Vibrio (Listonella) anguillarum. Despite a lot of research efforts, the virulence factors and mechanism of V. anguillarum are still insufficiently known, in part because of the lack of standardized virulence assays. METHODOLOGY/PRINCIPAL FINDINGS: We investigated and compared the virulence of 15 V. anguillarum strains obtained from different hosts or non-host niches using a standardized gnotobiotic bioassay with European sea bass (Dicentrarchus labrax L.) larvae as model hosts. In addition, to assess potential relationships between virulence and genotypic and phenotypic characteristics, the strains were characterized by random amplified polymorphic DNA (RAPD) and repetitive extragenic palindromic PCR (rep-PCR) analyses, as well as by phenotypic analyses using Biolog's Phenotype MicroArray™ technology and some virulence factor assays. CONCLUSIONS/SIGNIFICANCE: Virulence testing revealed ten virulent and five avirulent strains. While some relation could be established between serotype, genotype and phenotype, no relation was found between virulence and genotypic or phenotypic characteristics, illustrating the complexity of V. anguillarum virulence. Moreover, the standardized gnotobiotic system used in this study has proven its strength as a model to assess and compare the virulence of different V. anguillarum strains in vivo. In this way, the bioassay contributes to the study of mechanisms underlying virulence in V. anguillarum.
Subject(s)
Bass/microbiology , Genotype , Phenotype , Vibrio/genetics , Vibrio/pathogenicity , Animals , DNA, Bacterial/genetics , Larva/microbiology , Phylogeny , Polymerase Chain Reaction , Vibrio/classification , Vibrio/physiology , VirulenceSubject(s)
Bacteria/genetics , Beta vulgaris/microbiology , Food Preservation/methods , Industrial Microbiology/methods , Oligonucleotide Array Sequence Analysis/methods , Sucrose/metabolism , Bacteria/growth & development , Beta vulgaris/chemistry , Colony Count, Microbial/methods , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , Food Microbiology , Hydrogen-Ion Concentration , Kinetics , Osmolar Concentration , Population Dynamics , Species Specificity , Time FactorsABSTRACT
Tetragenococcus halophilus is known to flourish in extreme salt environments. Recently, this halophilic bacterium also appeared as the dominant microflora during storage of sugar thick juice, an intermediate product of beet sugar production. Although T. halophilus can cause degradation of thick juice, dominance of this bacterium does not always result in degradation. In this study T. halophilus strains from high-salt and high-sugar environments, and in particular from degraded and non-degraded thick juice, were compared in detail. Both physiological and genetic characterization using Biolog, repetitive PCR fingerprinting (rep-PCR) and random amplified polymorphic DNA (RAPD) technology, revealed clear differences between T. halophilus strains isolated from salt- and sugar-rich environments. However, no strain pattern could be specifically and systematically associated with degraded or non-degraded thick juice. Remarkably, halophilic T. halophilus strains were not able to grow in sugar thick juice. Irrespective of the differences between the strains from high-salt or high-sugar environments, DNA-DNA hybridization grouped all strains within the species T. halophilus, except one isolate from sugar thick juice that showed different physiological and genetic characteristics, and that may represent a new species of Tetragenococcus.
Subject(s)
Food Microbiology , Gram-Positive Cocci/genetics , Gram-Positive Cocci/metabolism , Sodium Chloride/metabolism , Sucrose/metabolism , Beta vulgaris/microbiology , DNA Fingerprinting , DNA, Bacterial/genetics , Genes, Bacterial , Genes, rRNA , Nucleic Acid Hybridization , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Random Amplified Polymorphic DNA TechniqueABSTRACT
The diversity of the bacterial population in sugar thick juice, an intermediate product in the production of beet sugar, which exhibits an extreme, osmophilic environment with a water activity value (a(w)) less than 0.86, was assessed with both culture-dependent and -independent 16S ribosomal RNA (rRNA) gene-based analyses. In comparison with previous studies, the number of different thick juice bacterial species increased from 29 to 72. Remarkably, a limited, gram-positive, culturable flora, encompassing species of Bacillus, Staphylococcus and mainly Tetragenococcus dominated thick juice during storage, while a more heterogeneous and unculturable fraction of Acinetobacter, Sporolactobacillus and Thermus species could be detected in freshly produced thick juice. Notably, almost all bacteria detected in the thick juice were also detected in the air, emphasising the importance of further investigation and assessment of strategies to reduce (air) contamination during processing and storage. The discovery of the contamination source may be used for the development of management strategies for thick juice degradation resulting from microbial activity.
