Éditeur(s) : Elsevier
Résumé : Because Vibrio aestuarianus is known to cause serious infections in Pacific oyster Crassostrea gigas, a real-time PCR assay was developed targeting the dnaJ gene of this bacterium. Only V. aestuarianus strains isolated from C. gigas mortality events in different geographic areas and the reference strain tested positive, whereas no amplification products was obtained with type strains belonging to 23 other species of Vibrio. Sensitivity and reproducibility of the method were assessed using either seawater or oyster homogenate samples spiked with one V aestuarianus strain. All these samples were stored at -20 degrees C in order to mimic retrospective or grouped natural sample analysis without quantification bias due to prolonged freezing. Analysis of standard curves revealed excellent correlation values between light microscopy cell enumerations and PCR Threshold Cycle (Ct) values, and acceptable PCR reaction efficiencies for all type of samples. Quantification curves of both sample types were equivalent, with a detection level as low as 1.6 V. aestuarianus cells in the PCR reaction tube, corresponding to 1.6.10(2) cells ml(-1) and 1.6.10(2) cells mg(-1) in seawater and entire oyster samples, respectively, taking into account the dilution factor used for appropriate template DNA preparation. Comparison of PCR assay reproducibility according to the complexity of samples revealed that seawater samples gave more reproducible quantification measures than samples from oyster homogenate, with precision of measured Ct values inferior to 0.4 and 0.6 respectively at 99% confidence. Use of the real-time PCR assay allowed us to monitor V. aestuarianus load in oysters naturally infected with this pathogen. Furthermore, we were able to detect V aestuarianus in samples of seawater in which oysters had been reared and in algal cultures used for feeding oysters. Because of the rapidity and reliability of the real-time PCR assay method used in this study, just a few hours are needed compared with the two days required using the classic culture method, this technique will be particularly valuable in mollusc pathology laboratories, for monitoring the source and course of infections by V. aestuarianus in pathogenesis and epidemiologic studies, as well as for designing appropriate prophylactic control measures. (C) 2009 Elsevier B.V. All rights reserved.
Journal of Microbiological Methods (0167-7012) (Elsevier), 2009-05 , Vol. 77 , N. 2 , P. 191-197

Droits : 2009 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/2009/publication-6447.pdf
DOI:10.1016/j.mimet.2009.01.021
http://archimer.ifremer.fr/doc/00000/6447/

Éditeur(s) : Walter De Gruyter & Co
Résumé : The section of the Liguro-Provencal basin north of the Balearic Islands is one of the most productive sites in the whole Mediterranean Sea, with intense phytoplankton bloom lasting about 2 months in late winter-early spring. The phytoplankton species composition of the area was investigated using light and electron microscopy to analyze bottle and net samples collected at several stations in spring 2000 and 2003. Serial dilution cultures established from bottle samples were also examined. A total of 168 phytoplankton taxa was identified, consisting of 73 Coscinodiscophyceae, 47 Dinophyceae, 25 Coccolithophyceae and 4 Prymnesiophyceae, 5 Prasinophyceae, 3 Chrysophyceae, 2 Cryptophyceae, 2 Dictyochophyceae, 1 Euglenophyceae, 1 Pelagophyceae, 1 Choanoflagellidea, 1 Filosea and 3 incertae sedis. We also provided a brief taxonomic description and original micrographs for 25 of the smallest and/or less known species identified in the study area, which may go undetected during routine microscopical analysis of fixed samples. Among these, 10 species were recorded for the first time in the Mediterranean Sea, confirming the need of detailed studies to reveal the biodiversity and biogeography of Mediterranean phytoplankton.
Botanica Marina (0006-8055) (Walter De Gruyter & Co), 2011-06 , Vol. 54 , N. 3 , P. 243-267

Droits : 2011 by Walter de Gruyter
http://archimer.ifremer.fr/doc/00038/14916/12245.pdf
DOI:10.1515/BOT.2011.033
http://archimer.ifremer.fr/doc/00038/14916/

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
The temperate symbiotic sea anemone Anemonia viridis, a member of the Cnidaria phylum, is a relevant experimental model to investigate the molecular and cellular events involved in the preservation or in the rupture of the symbiosis between the animal cells and their symbiotic microalgae, commonly named zooxanthellae. In order to increase research tools for this model, we developed a primary culture from A. viridis animal cells. By adapting enzymatic dissociation protocols, we isolated animal host cells from a whole tentacle in regeneration state. Each plating resulted in a heterogeneous primary culture consisted of free zooxanthellae and many regular, small rounded and adherent cells (of 3-5 mu m diameter). Molecular analyses conducted on primary cultures, maintained for 2 weeks, confirmed a specific signature of A. viridis cells. Further serial dilutions and micromanipulation allowed us to obtain homogenous primary cultures of the small rounded cells, corresponding to A. viridis ``epithelial-like cells''. The maintenance and the propagation over a 4 weeks period of primary cells provide, for in vitro cnidarian studies, a preliminary step for further investigations on cnidarian cellular pathways notably in regard to symbiosis interactions.
ISSN: 0920-9069

hal-01546173
https://hal.archives-ouvertes.fr/hal-01546173
DOI : 10.1007/s10616-013-9566-2