Éditeur(s) : Wiley-blackwell
Résumé : Aims To describe the diversity of the culturable mesophilic and potentially pathogenic vibrios isolated at 22 and 37 degrees C on TCBS medium, in September 2009 from seawater and surface sediments. Methods and Results q-PCR assays previously selected for the identification of bacterial strains isolated at 37 degrees C were used in combination with the partial sequencing of two housekeeping genes, pyrH and toxR, to identify 315 strains isolated at 22 degrees C. The great majority of the 37 degrees C strains was identified by q-PCR assays, (five of the six species) with the predominance of Vibrio alginolyticus (85 center dot 9%) and V.harveyi (10 center dot 7%). The human pathogens V.parahaemolyticus and V.cholerae were rarely detected (two strains each). The 22 degrees C strains were successfully identified by the phylogeny analysis of pyrH and toxR genes, revealing 20 Vibrio species, with the predominance of the clam pathogen V.celticus (36 center dot 8%). The Splendidus and the Harveyi groups represented the main Vibrio group at 22 degrees C (80%) and 37 degrees C (99 center dot 5%), respectively. Conclusions The combination of q-PCR assays and the sequencing of pyrH and toxR genes highlighted two different Vibrio communities at 22 and 37 degrees C both dominated by pathogenic species for marine organisms. Significance and Impact of the Study The sequencing of the pyrH gene revealed to be a valuable tool to identify environmental Vibrio spp. strains isolated at 22 degrees C, as 92 center dot 3% of them were identified in this study.
Journal Of Applied Microbiology (1364-5072) (Wiley-blackwell), 2013-06 , Vol. 114 , N. 6 , P. 1713-1724

Droits : 2013 The Society for Applied Microbiology
http://archimer.ifremer.fr/doc/00144/25502/24832.pdf
DOI:10.1111/jam.12181
http://archimer.ifremer.fr/doc/00144/25502/

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/

Résumé : Les bactéries appartenant au genre Vibrio sont largement répandues en milieu côtier tropical et tempéré. Certaines espèces sont pathogènes pour l'homme, comme Vibrio cholerae 01 et 0139 responsables du choléra, ou V. cholerae non-01/non-0139, V. parahaemolyticus et V. vulnificus impliqués dans de nombreuses pathologies (gastro-entérites, des septicémies et autres infections). En France, le risque associé à la présence de vibrions en milieu marin est mal évalué. Ce rapport fait le bilan des données disponibles sur la contamination littorale et les infections à vibrions en France. Une large place a été consacrée aux études écologiques, réalisées sur les sites d'implantation des centrales nucléaires sur le littoral français. Dans la population, le nombre d'infections est restreint car seules les pathologies graves sont recensées. Il s'agit de cas isolés, le plus souvent immuno-déprimés, enregistrés en été et en automne. Le milieu marin (par contact, baignade ou consommation de produits de la mer) est très souvent cité comme source probable de la contamination. Un petit nombre d'espèces de vibrions est concerné (V. alginolyticus, V. vulnificus, V. parahaemolyticus, V. cholerae non-01/non-0139 et V. hollisae). Dans l'environnement marin, l'espèce dominante est V. alginolyticus ; V. vulnificus et V. parahaemolyticus sont détectés sporadiquement. Le suivi, pendant plus de dix ans, des effets d'un réchauffement artificiel des eaux littorales à Gravelines a souligné des modifications sur les populations de Vibrio marins (présence des Vibrio en hiver, augmentation de la densité bactérienne et de la diversité spécifique). Le développement de techniques de biologie moléculaire devrait permettre une meilleure évaluation du risque sanitaire en quantifiant et en caractérisant les vibrions présents en milieu côtier (stade viable non cultivable, maintien du pouvoir pathogène et virulence des souches présentes).

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00104/21545/19124.pdf
DOI:Ifremer 2002
http://archimer.ifremer.fr/doc/00104/21545/

Résumé : Seawater, algae, micro-diets and Artemia are the main inputs in shrimp larval rearing. Each of them are potential carriers of pathogenic bacteria in the rearing media and weaken the animal in its early life stages. Daily monitoring of rearing water and larvae showed a bacterial spike after the first live feed meal. Vibrio settlement can be enhancedby giving Artemia with their incubation water where vibrio concentration can reach up to 105 CFU per milliliter. This study showed that simple measures such as rinsing and supplying Artemia with freshwater allowed a significant drop in their bacterial load.
L’élevage larvaire de crevettes nécessite l’utilisation de plusieurs intrants que sont l’eau de mer, les algues, les micro-particules et les Artemia. Chacun de ces intrants peut être une porte d’entrée potentielle de bactéries pathogènes et affaiblir l’animal dans les stades précoces. Des analyses bactériennes de l’eau d’élevage et des larves au quotidien montrent que c’est après l’introduction des proies vivantes que la flambée bactérienne est observée. La présence de vibrio peut être favorisée par la distribution des artémies dans leur eau d’incubation où il a pu être dénombré jusqu’à 105 CFU/ml. Cette fiche biotechnique montre qu’une mesure simple et efficace telle que le rinçage et la distribution des proies vivantes dans l’eau douce permet de réduire significativement la quantité de bactéries apportée par les Artemia.

Droits : 2008 Ifremer
http://archimer.ifremer.fr/doc/00118/22971/20801.pdf
http://archimer.ifremer.fr/doc/00118/22971/

Éditeur(s) : Elsevier
Résumé : We studied the effects of a lactic acid bacterium, Pediococcus acidilactici (strain MA 18/5M, CNCM), as a dietary probiotic on growth performance and some nutritional and microbiological aspects of the shrimp Litopenaeus stylirostris. Experimental shrimp culture was carried out over 10 weeks, using floating cages of 14 m(2) each set in earthen ponds, in a New Caledonia farm affected by "Summer syndrome", a septicaemic vibriosis caused by Vibrio nigripulchritudo. The experiment design testing probiotic-coated pellets against control was replicated in two earthen ponds. High mortalities, characteristic of Summer syndrome, were observed during the first 2 weeks of the trial. The probiotic improved production in the treated cages from both ponds with increases in the survival rate (7% and 15% respectively) and final biomass (8% and 12% respectively). No differences were recorded among treatments on growth performances, but significant lower food conversion ratios were obtained with probiotic treatment. After 5 weeks of rearing, the Hepatosomatic Index and the adjusted dry weight of the digestive gland were significantly increased by 10% and by 9% respectively in shrimps treated with probiotic. In the meantime, the specific activities of alpha amylase and trypsin in the digestive gland showed a significant effect of the probiotic treatment with respective increases by 35% and 55%. The rise in total trypsin activity following morning feeding was also enhanced by the probiotic treatment (P < 0.001). The highest concentration of probiotic (between 10(4)-10(5) CFU g(-1) of fresh gut) in the shrimp gut was obtained 2 h after feeding. The concentration remained high for 4 h after feeding before decreasing until the next meal. Total bacterial counts on Marine agar and TCBS in the gut were significantly lowered by the probiotic treatment over the 10 weeks of the experiment. Additionally, during the first 2 weeks, prevalence and load of V nigripulchritudo strains in haemolymph was lower in animals fed with the probiotic diet. This study demonstrated, under pond conditions, that feeding shrimp with live terrestrial lactic acid bacteria can be an effective treatment for improving shrimp culture affected by vibriosis. (c) 2008 Elsevier B.V. All rights reserved.
Aquaculture (0044-8486) (Elsevier), 2008-03 , Vol. 275 , N. 1-4 , P. 182-193

Droits : 2008 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/2008/publication-4079.pdf
DOI:10.1016/j.aquaculture.2008.01.011
http://archimer.ifremer.fr/doc/00000/4079/

Éditeur(s) : HAL CCSD Nakanishi Printing Co
Résumé : International audience
In Vibrio cholerae, the etiological agent of cholera, most of the virulence genes are located in two pathogenicity islands, named TCP (Toxin-Co-regulated Pilus) and CTX (Cholera ToXins). For each V. cholerae pathogenicity gene, we retrieved every primer published since 1990 and every known allele in order to perform a complete in silico survey and assess the quality of the PCR primers used for amplification of these genes. Primers with a melting temperature in the range 55-60 degrees C against any target sequence were considered valid. Our survey clearly revealed that two thirds of the published primers are not able to properly detect every genetic variant of the target genes. Moreover, the quality of primers did not improve with time. Their lifetime, i.e. the number of times they were cited in the literature, is also not a factor allowing the selection of valid primers. We were able to improve some primers or design new primers for the few cases where no valid primer was found. In conclusion, many published primers should be avoided or improved for use in molecular detection tests, in order to improve and perfect specificity and coverage. This study suggests that bioinformatic analyses are important to validate the choice of primers.
ISSN: 1342-6311

hal-01546178
https://hal.archives-ouvertes.fr/hal-01546178
DOI : 10.1264/jsme2.ME11317

Éditeur(s) : Amer Soc Microbiology
Résumé : A molecular typing study on Vibrio strains implicated in shrimp disease outbreaks in New Caledonia and Japan was conducted by using AP-PCR (arbitrarily primed PCR). It allowed rapid identification of isolates at the genospecies level and studies of infraspecific population structures of epidemiological interest. Clusters identified within the species Vibrio penaeicida were related to their area of origin, allowing discrimination between Japanese and New Caledonian isolates, as well as between those from two different bays in New Caledonia separated by only 50 km. Other subclusters of New Caledonian V. penaeicida isolates could be identified, but it was not possible to link those differences to accurate epidemiological features. This contribution of AP-PCR to the study of vibriosis in penaeid shrimps demonstrates its high discriminating power and the relevance of the epidemiological information provided. This approach would contribute to better knowledge of the ecology of Vibrio spp. and their implication in shrimp disease in aquaculture.
Applied And Environmental Microbiology (0099-2240) (Amer Soc Microbiology), 1999-03 , Vol. 65 , N. 3 , P. 1145-1151

Droits : American society for microbiology
http://archimer.ifremer.fr/doc/1999/publication-2734.pdf
http://archimer.ifremer.fr/doc/00000/2734/

Éditeur(s) : Academic Press Ltd- Elsevier Science Ltd
Résumé : Extracellular products (ECPs) of the pathogenic Vibrio aestuarianus 01/32 were previously reported to display lethality in Crassostrea gigas oysters and to cause morphological changes and immunosuppression in oyster hemocytes. To identify the source of this toxicity, biochemical and genetic approaches were developed. ECP protease activity and lethality were shown to be significantly reduced following incubation with metal chelators, suggesting the involvement of a zinc metalloprotease. An open reading frame of 1836 bp encoding a 611-aa metalloprotease (designated yam) was identified. The deduced protein sequence showed high homology to other Vibrio metalloproteases reported to be involved in pathogenicity. To further confirm the role of this enzyme in ECP toxicity, a plasmid carrying the yarn gene under the control of an araC-P-BAD expression cassette was transferred to a Vibrio splendidus related strain, LMC20012(T), previously characterized as non-pathogenic to oysters. Expression of Vam conferred a toxic phenotype to LMG20012(T) ECPs in vivo and cytotoxicity to oyster hemocytes in vitro. Collectively, these data suggest that the Vam metalloprotease is a major contributor to the toxicity induced by V aestuarianus ECPs and is involved in the impairment of oyster hemocyte functions. (C) 2010 Elsevier Ltd. All rights reserved.
Fish & Shellfish Immunology (1050-4648) (Academic Press Ltd- Elsevier Science Ltd), 2010-11 , Vol. 29 , N. 5 , P. 753-758

Droits : 2010 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/00015/12649/9588.pdf
DOI:10.1016/j.fsi.2010.07.007
http://archimer.ifremer.fr/doc/00015/12649/

Éditeur(s) : Elsevier
Résumé : In a previous study, we demonstrated the existence of an emerging cluster of Vibrio nigripulchritudo that proved to be associated with shrimp mortality events in New Caledonia. Using sequence polymorphisms evidenced in this previous MultiLocus Sequence Typing study, we developed two new quantitative PCR assays permitting the detection and quantification of V. nigripulchritudo at the genospecies level using SYBR Green I chemistry and at the emerging cluster level using Fluorescence Resonance Energy Transfer technology with hybridization probes. The use of this molecular diagnostic tool evidenced the colonization of the shrimp pond ecosystem by the pathogenic cluster at least at the onset of the disease. This new tool will allow better investigation of the dynamics of this bacterial pathogen in the shrimp farm ecosystem. (C) 2007 Elsevier B.V. All rights reserved.
Journal of Microbiological Methods (0167-7012) (Elsevier), 2007-07 , Vol. 70 , N. 1 , P. 30-38

Droits : 2007 Elsevier B.V. All rights reserved
http://archimer.ifremer.fr/doc/2007/publication-2729.pdf
DOI:10.1016/j.mimet.2007.03.007
http://archimer.ifremer.fr/doc/00000/2729/

Éditeur(s) : Universidad Autonoma de Baja California
Résumé : The pathogenicity of two V. penaeicida strains, AM101 and KH-1, with different geographic origin, and V. nigripulchritudo strain AM102, were investigated in juvenile blue shrimp species Litopenaeus stylirostris. Alive bacteria and protein fractions (PFs) obtained from cell-free supernatants (CFS) were used in experimental challenges. Strains AM102, AM101, and KH-1 produced respectively 60, 54 and 12% mortality at 96h after infection using 104 cfu mL-1 of bacterial suspension. Exocellular toxin-like factors were evidenced in CFS from the New Caledonian strains (AM102 and AM101) but not the Japanese strain (KH-1). At 48h post injection of each CFS, mortality rates were respectively 96,98 and 5% when these strains were cultivated at 20°C, whereas only O,16 and 5% mortality rates were observed when these strains were cultivated at 30°C. Clear differences in pathogenicity between both V. penaeicida strains of distinct geographic origin (AM101 and KH-1) were thus emphasized. Protein fractions were obtained from CFS of all the strains tested in this study and cultivated at 20°C, by ammonium sulphate precipitation. Whatever the used strain, significantly higher mortalities were produced with PFs obtained with 0-40% of ammonium sulphate saturation, in respect to those produced with PFs60 and PFs80. Shrimp injected with PFs40 from strains AM102, AM101 and KH-1, and at a 20-µg org-1 dose produced respectively 100, 90 and 60% mortality 46h after the challenge. The strain AM101 showed a mediam lethal dose of approximately 5µg protein org-1 (1-1.25 µg protein g-1 body weight) 59h after injection. The PFs40 from New Caledonian strains were fond sensitive to heating and proteinase K treatments, reinforcing thus the hypothesis of their proteinaceous nature. Surprisingly, PFs40 from each bacterial strain displayed similar protein bands by SDS-PAGE suggesting that the tested strains share a common exotoxic compound regardless their distinct geographic origin or species.
Ciencias Marinas (Universidad Autonoma de Baja California), 2003 , Vol. 29 , N. 1 , P. 77-88

Droits : Universidad Autonoma de Baja California
http://archimer.ifremer.fr/doc/2003/publication-2807.pdf
http://archimer.ifremer.fr/doc/00000/2807/

Éditeur(s) : Elsevier Science Bv
Résumé : In tropical shrimp ponds, the increasing of feed input, concomitantly with the stocking shrimp biomass, induces an eutrophication of the ecosystem. Although difficult to maintain, its stability is required to guarantee the success of the culture. A 110-day period of phytoplankton and bacterioplankton stock and dynamics in an earthen pond (1.2 ha area, 1 m depth) was monitored using flow cytometry to provide baseline information on community characteristics and ecosystem instability. Seven autotrophic cell types were identified over the whole sampling period. Prokaryotic cells included Synechococcus sp., a group named UNK which presented an atypical new flow cytometric signature and picoeukaryotes (PEUK). Nanophytoplankton cells were represented by 4 groups: NAN1, NAN2, NAN3 and Cryptophytes. During the first part of the survey, picophytoplankton dominated the phytoplanktonic assemblage. The mean abundance of total cells (up to 8 x 10(6) cells mL(-1)) was among the highest recorded in marine and brackish waters. Bacterial abundance and production ranged from 0.8 to 5.1 x 10(7) cells mL(-1) and from 30 to 110 mu g C L-1 h(-1). A shift from pico to nanophytoplankton abundance was observed for a few days from d 96. During this period, heterotrophic bacteria production and abundance suddenly dropped, implying a change in the functioning of the microbial loop. This shift was concomitant with a significant shrimp mortality outbreak due to Vibrio penaeicida, the etiological agent of a disease known as Syndrome 93, which affects the shrimp industry in New Caledonia. This survey suggests that flow cytometric analysis could be used for the monitoring of aquaculture systems to improve our understanding of the complex phytoplankton and bacterial dynamics of these systems and its potential influence on disease development. (c) 2010 Elsevier B.V. All rights reserved.
Aquaculture (0044-8486) (Elsevier Science Bv), 2010-12 , Vol. 310 , N. 1-2 , P. 112-121

Droits : 2010 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/00029/14068/11312.pdf
DOI:10.1016/j.aquaculture.2010.10.022
http://archimer.ifremer.fr/doc/00029/14068/

Éditeur(s) : Academic Press Ltd- Elsevier Science Ltd
Résumé : Antioxidant defences and induced oxidative stress tissue damage of the blue shrimp Litopenaeus stylirostris, under challenge with Vibrio nigriputchritudo, were investigated for a 72-h period. For this purpose, L stylirostris were first infected by immersion with pathogenic V. nigripulchritudo strain SFn1 and then antioxidant defences: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (Gpx), Total antioxidant status (TAS), glutathiones and induced tissue damage (MDA and carbonyl proteins) were determined in the digestive gland at 0, 12, 24, 48 and 72 h post-infection (h.p.i.). In the meantime, TAS was also measured in the blood. Infection level of the shrimps during the challenge was followed by determining V. nigripulchritudo prevalence and load in the haemolymph of the shrimps. Changes in all these parameters during the 72-h.p.i. period were recorded for control shrimps and shrimps previously fed for one month with probiotic Pediococcus aciditactici MA18/5M at 10(7) CFU g(-1) of feed. Our results showed that immersion with V nigripulchritudo led to maximal infection level in the haemolymph at 24 h.p.i. preceding the mortality peak recorded at 48 h.p.i. Significant decreases in the antioxidant defences were detected from 24 h.p.i. and beyond that time infection leaded to increases in oxidative stress level and tissue damage.. Compared to control group, shrimps fed the probiotic diet showed lower infection (20% instead of 45% at 24 h.p.i. in the control group) and mortality (25% instead of 41.7% in the control group) levels. Moreover, infected shrimp fed the probiotic compared to uninfected control shrimps exhibited very similar antioxidant status and oxidative stress level. Compared to the infected control group, shrimps fed the probiotic sustained higher antioxidant defences and lower oxidative stress level. This study shows that bacterial infection leads to oxidative stress in L. stylirostris and highlighted a beneficial effect of P. acidilactici, suggesting both a competitive exclusion effect leading to a reduction of the infection level and/or an enhancement of the antioxidant status of the shrimps. (C) 2010 Elsevier Ltd. All rights reserved.
Fish & Shellfish Immunology (1050-4648) (Academic Press Ltd- Elsevier Science Ltd), 2010-04 , Vol. 28 , N. 4 , P. 622-631

Droits : 2010 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/00003/11419/7984.pdf
DOI:10.1016/j.fsi.2009.12.024
http://archimer.ifremer.fr/doc/00003/11419/

Éditeur(s) : Wiley-blackwell
Résumé : Aims Exopolysaccharides (EPS) are industrially valuable molecules with numerous useful properties. This study describes the techniques used for the identification of a novel Vibrio bacterium and preliminary characterization of its EPS. Methods and Results Bioprospection in marine intertidal areas of New Caledonia followed by screening for EPS producing brought to selection of the isolate NC470. Phylogenetic analysis (biochemical tests, gene sequencing and DNADNA relatedness) permitted to identify NC470 as a new member of the Vibrio genus. The EPS was produced in batch fermentation, purified using the ultrafiltration process and analysed by colorimetry, Fourier Transform Infrared spectroscopy, gas chromatography, Nuclear Magnetic Resonance and HPLC-size exclusion chromatography. This EPS exhibits a high N-acetyl-hexosamines and uronic acid content with a low amount of neutral sugar. The molecular mass was 672x103Da. These data are relevant for possible technological exploitation. Conclusions We propose the name Vibrio neocaledonicus sp. nov for this isolate NC470, producing an EPS with an unusual sugar composition. Comparison with other known polymers permitted to select applications for this polymer. Significance and Impact of the Study This study contributes to evaluate the marine biodiversity of New Caledonia. It also highlights the biotechnological potential of New Caledonia marine bacteria.
Journal Of Applied Microbiology (1364-5072) (Wiley-blackwell), 2013-06 , Vol. 114 , N. 6 , P. 1702-1712

Droits : 2013 The Society for Applied Microbiology
http://archimer.ifremer.fr/doc/00144/25501/24036.pdf
DOI:10.1111/jam.12184
http://archimer.ifremer.fr/doc/00144/25501/

Résumé : The New-Caledonian shrimp industry is based on the controlled reproduction of the shrimp Litopenaeus stylirostris, a species which was introduced in the 80s. The major difficulty to which the industry has been faced for 10 years is the occurrence of the "syndrome 93", which corresponds to mortality phases when the temperature falls down in April-May-June. This mortality is associated to the pathogenic bacteria Vibrio penaecida and is expressed at different levels which are variable from year ta year and from pond to pond. No resistance to this pathology has been developed spontaneously. This is likely due to the protocole used to rear spawners, which does not allow to implement an efficient selective pressure at each generation An experimental selection on the criteria of survival after picks of syndrome 93 has been conducted at the Laboratoire Aquacole de Calédonie. The 3rd selected generation demonstrates survival rates improved by 20% during experimental infections with V. penaeicida in comparison with a non selected control population of same genetic origin. The comparison of the correlated responses on the level of expression of 5 genes which are potentially implicated in immunity phenomena (Peneidins, lysozyme, transglutaminase. profiline, annexine) shows that the selected population has a level of expression in lyzozyme twice higher than the control population. This result suggests that the lysozyme could be a genetic marker which could be used in a selective breeding program to he developed in relation with the private hatcheries.
La filière crevette de Nouvelle-Calédonie repose sur la maîtrise de la reproduction contrôlée de la crevette Litopenaeus stylirostris, espèce introduite dans les années 80. La difficulté majeure que rencontre la filière depuis une dizaine d'années est la récurrence du « syndrome 93 », qui correspond à des épisodes de mortalités lors des baisses de température en avril-mai-juin. Ces mortalités sont associées à la bactérie pathogène Vibrio penaeicida et s'expriment à des niveaux d'intensité variable d'une année à l'autre et d'un bassin à l'autre. Aucune résistance vis-à-vis de cette pathologie ne s'est développée spontanément. Ceci est vraisemblablement lié au protocole employé pour l'élevage des géniteurs qui ne permet pas d'exercer une pression de sélection efficace à chaque génération. Une expérience de sélection sur un critère de survie à des épisodes de syndrome 93 a été menée au Laboratoire Aquacole de Calédonie. La 3ème génération sélectionnée montre des survies améliorées de l'ordre de 20% lors d'infections expérimentales à V. penaeicida par rapport à une population témoin non sélectionnée de même origine génétique. La comparaison des réponses corrélées sur le niveau d'expression de 5 gènes potentiellement impliqués dans les phénomènes de défense immunitaire (penaeidine, lysozyme, transglutaminase, profiline, annexine) montre que la population sélectionnée a un niveau d'expression en lysozyme deux fois plus élevé que la population témoin. Ce résultat suggère que le lysozyme pourrait être un marqueur génétique utilisable dans un programme de sélection à développer en relation avec les écloseries de production.

Droits : 2003 Ifremer
http://archimer.ifremer.fr/doc/00109/22020/19643.pdf
http://archimer.ifremer.fr/doc/00109/22020/

Éditeur(s) : Springer
Résumé : Two recurring syndromes threaten the viability of the shrimp industry in New Caledonia, which represents the second largest export business. The "Syndrome 93" is a cold season disease due to Vibrio penaeicida affecting all shrimp farms, while the "Summer Syndrome" is a geographically restricted vibriosis caused by a virulent lineage of Vibrio nigripulchritudo. Microbiological procedures for diagnosis of these diseases are time-consuming and do not have the ability to discriminate the range of virulence potentials of V. nigripulchritudo. In this study, we developed a multiplex PCR method to simultaneously detect these two bacterial species and allow for pathotype discrimination. The detection limits of this assay, that includes an internal amplification control to eliminate any false-negative results, were determined at 10 pg purified DNA and 200 cfu/ml. After confirming the effectiveness of our method using experimentally infected animals, its accuracy was compared to standard biochemical methods during a field survey using 94 samples collected over 3 years from shrimp farms encountering mortality events. The multiplex PCR showed very high specificity for the detection of V. penaeicida and V. nigripulchritudo (inclusivity and exclusivity 100%) and allowed us to detect the spreading of highly pathogenic isolates of V. nigripulchritudo to a farm adjoining the "Summer Syndrome area." This assay represents a simple, rapid, and cost-effective diagnostic tool for implementing timely risk management decisions but also understanding the seasonal and geographical distribution of these pathogens.
Microbial Ecology (0095-3628) (Springer), 2012-01 , Vol. 63 , N. 1 , P. 127-138

Droits : Springer Science+Business Media, LLC 2011
http://archimer.ifremer.fr/doc/00063/17398/15169.pdf
DOI:10.1007/s00248-011-9951-3
http://archimer.ifremer.fr/doc/00063/17398/

Éditeur(s) : Elsevier Sci Ltd
Résumé : Diverse families of antimicrobial peptides and proteins have been described in oysters. We investigated here how antimicrobials are involved in the immune response against a pathogenic strain of Vibrio splendidus. Oyster antimicrobials were shown to display a wide variety of expression profiles in hemocyte populations and tissues. Oyster defensins are constitutively expressed in specific tissues such as mantle (Cg-Defm) or hemocytes (Cg-Defhs), while Cg-BPI is inducible and Cg-Prp appears down-regulated in hemocytes upon infection. The migratory behavior of hemocytes that express the different antimicrobials was found to be involved in the oyster response to a pathogenic Vibrio infection. Indeed, it contributes to colocalize several antimicrobials that were shown here to have synergistic activities. We propose that such a synergy, which was evidenced both within and between families of antimicrobials, might compensate for the low concentration of antimicrobials in oyster tissues. (c) 2012 Elsevier Ltd. All rights reserved.
Developmental And Comparative Immunology (0145-305X) (Elsevier Sci Ltd), 2012-07 , Vol. 37 , N. 3-4 , P. 363-370

Droits : 2012 Elsevier Ltd. All rights reserved.
http://archimer.ifremer.fr/doc/00093/20382/18267.pdf
DOI:10.1016/j.dci.2012.01.004
http://archimer.ifremer.fr/doc/00093/20382/

Éditeur(s) : Nature Publishing Group
Résumé : Vibrio nigripulchritudo is an emerging pathogen of farmed shrimp in New Caledonia and other regions in the Indo-Pacific. The molecular determinants of V. nigripulchritudo pathogenicity are unknown; however, molecular epidemiological studies have suggested that pathogenicity is linked to particular lineages. Here, we performed high-throughput sequencing-based comparative genome analysis of 16 V. nigripulchritudo strains to explore the genomic diversity and evolutionary history of pathogen-containing lineages and to identify pathogen-specific genetic elements. Our phylogenetic analysis revealed three pathogen-containing V. nigripulchritudo clades, including two clades previously identified from New Caledonia and one novel clade comprising putatively pathogenic isolates from septicemic shrimp in Madagascar. The similar genetic distance between the three clades indicates that they have diverged from an ancestral population roughly at the same time and recombination analysis indicates that these genomes have, in the past, shared a common gene pool and exchanged genes. As each contemporary lineage is comprised of nearly identical strains, comparative genomics allowed differentiation of genetic elements specific to shrimp pathogenesis of varying severity. Notably, only a large plasmid present in all highly pathogenic (HP) strains encodes a toxin. Although less/non-pathogenic strains contain related plasmids, these are differentiated by a putative toxin locus. Expression of this gene by a non-pathogenic V. nigripulchritudo strain resulted in production of toxic culture supernatant, normally an exclusive feature of HP strains. Thus, this protein, here termed 'nigritoxin', is implicated to an extent that remains to be precisely determined in the toxicity of V. nigripulchritudo.
Isme Journal (1751-7362) (Nature Publishing Group), 2013-10 , Vol. 7 , N. 10 , P. 1985-1996

Droits : 2013 International Society for Microbial Ecology All rights reserved 1751-7362/13
http://archimer.ifremer.fr/doc/00160/27117/28369.pdf
DOI:10.1038/ismej.2013.90
http://archimer.ifremer.fr/doc/00160/27117/

Éditeur(s) : Colloque IMBC - International Marine Biotechnology Conference
Résumé : Since 1997, a new pathology seasonally occurs in new caledonian shrimp farms during the warm season. Diseased Litopenaeus stylirostris shrimp suffer from a systemic vibriosis which was attributed to highly pathogenic Vibrio nigripulchritudo. At the present time, only two farms among 17 are affected by the so called "summer syndrome". In such a context it appears urgent to develop reliable diagnostic tools to detect V. nigripulchritudo strains and to differentiate highly virulent strains from non virulent ones in order to understand the mechanisms of infection and overcome disease spreading. Multilocus Sequencing Typing (MLST) analysis on 24 bacterial strains isolated in geographically distinct shrimp farms highlighted the existence of a cluster A containing only strains displaying moderate to high virulence. Since experimental infection procedure for virulence status determination is material intensive and time consuming, and MLST method not accurate enough to inform on the virulence status of strains, it appeared of great interest to develop other diagnostic tools. Two phylogenetically closed strains, one highly virulent SFn1 and the other non-virulent SFn118 were selected in order to characterize genes specific of the virulent strain by Substractive Suppression Hybridization (SSH). Firstly, a total of 622 DNA fragments specific of SFn1 were sequenced, characterized and further used as probes in a DNA macroarray. Testing 24 strains, hybridization profiles were found to be strongly correlated with both MLST genotypes and bacterial virulence status. A total of 96 representative probes were secondly selected to validate macroarray results on 34 more additionnal strains, allowing identification of genetic markers of virulence. Most of DNA fragments specific of highly virulent strains associated to summer syndrome were found to be bore by a plasmid successfully isolated in the laboratory. In next future, these results would permit the development of relevant diagnostic tools useful for accurate epidemiological survey of the summer syndrome.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2007/acte-3398.pdf
http://archimer.ifremer.fr/doc/00000/3398/

Éditeur(s) : INRA
Résumé : According to the world organisation for animal health (OIE) an emerging disease is defined as a recently admitted serious illness, whose aetiology can, or not, have already been established, and which is likely to be propagated within a population or between populations, for example at the time of international exchanges of aquatic animals and/or products of aquatic animals. Even though the emerging diseases that affect human health have been much studied, those which affect marine organisms and species of aquaculture interest in particular are poorly documented. By restricting emergence to only infectious diseases, we aim at presenting in a non-exhaustive way some scenarios of the emergence of the diseases of aquacultured species by illustrating them with three examples available in the scientific literature: one relating to the appearance of a pathogenic agent in a new host with the case of the herpesvirus of the Koï carp, the other with the evolution of a pre-existing pathogenic agent with the case of shrimp vibriosis due to Vibrio nigripulchritudo in New Caledonia, and the last example relating to the introduction of one pathogenic pre-existing pathogen in an unscathed area with the case of Bonamia ostreae infecting the flat oyster Ostrea edulis. The causes of the emergence of diseases are multiple and implicate in an intercurrent way pathogenic agents, the environment, the host or host species and anthropogenic factors. In the marine environment, these causes are very often ignored. In this context, the development of zoosanitary surveillance networks and diagnostic tools present a considerable interest in order to anticipate, prevent and/or intervene on the emergence of the diseases by limiting their sanitary, ecological and political consequences.
Selon l'Office International des Epizooties (OIE) une maladie émergente désigne une maladie grave récemment reconnue, dont la cause peut, ou non, avoir déjà été établie, et qui est susceptible de se propager au sein d'une population ou entre des populations, par exemple à l'occasion d'échanges internationaux d'animaux aquatiques et/ou de produits d'animaux aquatiques. Si les maladies émergentes qui affectent la santé humaine ont été très étudiées, celles qui touchent les organismes marins et les organismes aquacoles d'intérêt économique en particulier sont en revanche peu documentées. C'est en restreignant l'émergence aux seules maladies infectieuses que seront présentés de façon non exhaustive quelques scénarios de l'émergence des maladies chez les organismes d'intérêt aquacole en les illustrant par trois exemples disponibles dans la littérature scientifique : l'un relatif à l'apparition d'un agent pathogène chez un nouvel hôte avec le cas de l'herpesvirus de la carpe Koï, l'autre à l'évolution d'un agent pathogène existant avec le cas de la vibriose à Vibrio nigripulchritudo sévissant dans les élevages de crevettes pénéides de Nouvelle-Calédonie et enfin le dernier lié à l'introduction d'un pathogène préexistant avec le cas de Bonamia ostreae infectant l'huître plate Ostrea edulis. Les causes d'émergence de maladies sont multiples et font intervenir de façon intercurrente l'agent pathogène, l'environnement, l'hôte ou les espèces hôtes et des facteurs anthropiques. Dans le milieu marin, ces causes sont bien souvent méconnues. Dans ce contexte le développement des réseaux de surveillance et des techniques de diagnostic revêtent un intérêt considérable afin d'anticiper, de prévenir et/ou d'intervenir sur l'émergence des maladies en limitant leur conséquences sanitaires, écologiques et politiques.
INRA Productions Animales (INRA), 2007-07 , Vol. 20 , N. 3 , P. 207-212

Droits : INRA
http://archimer.ifremer.fr/doc/2007/publication-2980.pdf
http://archimer.ifremer.fr/doc/00000/2980/

Éditeur(s) : Elsevier
Résumé : Shrimp farming is a small but growing industry in New Caledonia. Since 1993, "Syndrome 93" has been affecting New Caledonian shrimp farming industry every cold season, causing severe epizootic mortalities in grow-out ponds and significant losses. Highly pathogenic strains of Vibrio penaeicida are considered the etiological agent of the disease in Litopenaeus stylirostris. On one hand, studies demonstrated that healthy shrimp may carry V penaeicida for weeks with a high overall prevalence, regardless of any seasonal pattern or temperature conditions. On the other hand, larvae are free of V penaeicida and are also resistant to experimental infection. V penaeicida is frequently detected in incoming water pumped from the bays, which was shown, by a molecular typing study, to be the infectious source. This particular epidemiological pattern highlights the major role of the factors that trigger and aggravate the disease in grow-out ponds, where shrimp populations carry the pathogen all year round. In order to gain a better understanding of "Syndrome 93" epidemiology, quantification of V penaeicida both in shrimp and the shrimp farm ecosystem is necessary. This article describes the steps in the successful development of a real-time PCR quantification assay of V penaeicida in shrimp haemolymph, seawater (from ponds or bays) and sediment pore water, including the choice of an accurate extraction technique. The entire detection method; including sample processing, DNA extraction and real-time PCR amplification, can be completed within 4 h. (c) 2006 Elsevier B.V All rights reserved.
Journal of Microbiological Methods (0167-7012) (Elsevier), 2006-10 , Vol. 67 , N. 1 , P. 27-35

Droits : 2006 Elsevier
http://archimer.ifremer.fr/doc/2006/publication-1903.pdf
DOI:10.1016/j.mimet.2006.02.013
http://archimer.ifremer.fr/doc/00000/1903/