In this paper we report the first characterization of cultivable bacteria obtained from the Antarctic sea urchin Sterechinus neumayeri. The coelomic fluid was obtained from a pool of sea urchins which was plated onto different media to isolate the bacteria. A total of 42 isolates of psychrotrophic and aerobic γ-Proteobacteria (59.5%), Flavobacteria (33.3%) and Actinomycetes (7.2%) were isolated and sequenced. These bacteria were exposed to heavy metals and antibiotics, where 38 strains were analysed by the minimal inhibitory concentration method. Antibiotic resistance was detected in 44% of cultivable strains, and a further 13% presented co-resistance to antibiotics and heavy metals. The genera of bacteria that showed an increased resistance and co-resistance to metals and antibiotics were Flavobacterium, Psychrobacter and Pseudomonas. Additionally, 30.9% of isolated bacterial strains contained plasmids, which are probably related to resistance and co-resistance to metals. These results indicate that sea urchin-associated bacteria could be reservoirs for antibiotic resistance genes.
Abstract.-The Patagonian toothfish Dissostichus eleginoides is one of the most important fisheries from the Southern Ocean. The biology of this species is relatively well studied and some nutritionals issues have also been reported; however there is no information about the composition of the bacterial community of the gastrointestinal tract, which is essential to characterize the microbiota of this fish. The bacterial flora of D. eleginoides is here described for the first time using culturable methods. By applying traditional culture-based techniques and 16S rDNA sequencing methods it was possible to characterize the families Vibronaceae and Moraxellaceae, which were mainly represented by Vibrio and Psychrobacter, respectively. This Patagonian fish shows a microbiota very similar to other cold waters fishes.
Two Vibrio strains (VPAP36 and VPAP40) were isolated from moribund-settled larvae of the Chilean scallop Argopecten purpuratus during vibriosis outbreaks that occurred in two commercial scallop larvae hatcheries located in the Inglesa and Tongoy bays in Northern Chile. The strains were identified as Vibrio chagasii using phenotypic characterization and whole genome sequence analysis. Both strains exhibited the phenotypic properties associated with virulence, gelatin hydrolysis and β-hemolysis, whereas only VPAP36 produced phospholipase and only VPAP40 produced caseinase. The whole genome analysis showed that the strains harbored genes encoding for the virulence factors, the EPS type II secretion system, and Quorum Sensing (auto-inductor 1 and auto-inductor 2), whereas genes encoding a metalloproteinase and a capsular polysaccharide were detected only in the VPAP40 genome. When challenge bioassays using healthy 11-day-old scallop larvae were performed, the V. chagasii VPAP36 and VPAP40 strains exhibited significant (p < 0.05) differences in their larval lethal activity, producing, after 48 h, larval mortalities of 65.51 ± 4.40% and 28.56 ± 5.35%, respectively. Otherwise, the cell-free extracellular products of the VPAP36 and VPAP40 strains produced larval mortalities of 20.86 ± 2.40% and 18.37 ± 2.40%, respectively, after 48 h of exposure. This study reports for the first time the isolation of V. chagasii from the massive larval mortalities of the farmed scallop (Argopecten purpuratus) in Chile, and demonstrates the pathogenic activity of V. chagasii towards the Chilean scallop, the second most important species for Chilean mariculture.
ABSTRACT.One of the main problems facing Atlantic halibut hatcheries is the high mortality in the early stages of larval development. Several factors could be involved, for example: water quality, diseases or abnormalities, such as deformities occurring in the yolk sac larvae prior to exogenous feeding. The aim of this study was to identify differences in bacterial flora associated with yolk sac larvae with oral deformity. We also aimed to establish whether there is any relationship between bacterial strains and the "gaping jaws" syndrome. During our study, 74 bacterial isolates were obtained using three different nutrient media: Marine Agar, R2A and TCBS. Some of these bacteria were characterized using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) and 16S rRNA sequencing. The immune response in larvae exhibiting the "gaping jaws" condition was measured by real time PCR. Our results showed significant differences in bacterial flora between normal and gaping larvae. The gaping yolk sac larvae were predominantly colonized by members of the families Vibrionaceae and Flavobacteriaceae. Bacteria belonging to the Bacillus and Pseudoalteromonas genera were also present but less frequent. It was not possible to associate a type or group of bacteria directly related to "gaping". Strikingly, larvae with gaping jaws had an increase in the expression of two immune related genes, like hepcidin and chemokine (MIP-1ß). These results indicate activation of the immune response in larvae with "gaping jaws" syndrome and this response could be related to bacteria isolated from gaping condition. Keywords: Atlantic halibut, Hippoglossus hippoglossus, bacterial flora, "gaping jaws", immune genes.Caracterización de la flora cultivable y la respuesta inmune en larvas con saco vitelino del halibut del Atlántico (Hippoglossus hippoglossus L.) con el síndrome de "gaping jaws" RESUMEN. Uno de los mayores problemas que enfrentan los criaderos de halibut del Atlántico es la alta mortalidad en estados tempranos del desarrollo larval. Distintos factores pueden estar involucrados, e.g., calidad del agua, enfermedades o anormalidades tales como deformidades que ocurren en las larvas en estado de saco vitelino antes de la alimentación exógena. El objetivo de este estudio fue identificar diferencias en la flora bacteriana asociada a larvas con saco vitelino con deformidad oral. Además se buscó establecer si existió relación entre alguna cepa bacteriana y el síndrome de "gaping jaws". Durante el estudio, 74 cepas bacterianas fueron aisladas usando diferentes medios de cultivo: agar marino, R2A y agar TCBS. Algunas de las bacterias fueron caracterizadas usando reacción en cadena de la polimerasa con análisis del polimorfismo de los fragmentos de restricción (PCR-RFLP) y secuenciación del gen ribosomal 16S. La respuesta inmune de larvas con el síndrome de "gaping jaws" fue medida por PCR en tiempo real. Los resultados evidenciaron diferencias significativas en la flora entre larvas normales y "gaping jaws". Las larvas c...
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