Éditeur(s) : PSIC 11 - 11th Pacific Science Inter-Congress : Pacific Countries and their Ocean: Facing Local and Global Changes / 2nd Symposium on French Research in the Pacific. March 2 - 6, 2009 Tahiti, French Polynesia.
Résumé : Experimental infections together with epidemiological studies have shown that pathogenic and non‐pathogenic isolates of V. nigripulchritudo co‐existed in shrimp farm environment (2, 3). Moreover, obtained results also demonstrated that the “summer syndrome”was caused by a single, possibly emerging, cluster of virulent strains. Consequently, it was hypothesized that pathogenic strains of V. nigripulchritudo may persist from one year to the next in the shrimp farm environment and re‐develop inside the grow‐out system at the following rearing cycle (3). This study was therefore aimed at determining whether V. nigripulchritudo isolates may survive, or not, in a shrimp pond bottom soil during a 18‐week drying period. To this end, V.nigripulchritudo mapping was performed with recently developed molecular tools and classical culture‐dependent techniques.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00197/30874/29242.pdf
http://archimer.ifremer.fr/doc/00197/30874/

Résumé : Ce document constitue le bilan détaillé des résultats du projet DEDUCTION (2007-2010) , action par action. Le projet DEDUCTION "DEveloppement DUrable de la Crevetticulture, Traitement de l’Information et Observatoire du système en Nouvelle-Calédonie" est un projet de recherche et de développement en soutien à la filière crevetticole néo-calédonienne. C'est un programme très horizontal et pluridisciplinaire en poursuivant et développant les ouvertures scientifiques, techniques et thématiques établies à la faveur de DESANS. Son ossature s’articule autour de 4 actions : 3 actions de recherche et une action transversale devant faciliter les interfaces entre la recherche et sa finalité, entre les chercheurs et les partenaires locaux (GFA, Provinces, Gouvernement, Haut-Commissariat), en intégrant les résultats "ferme" à l'échelle de la filière. Une tâche de « Communication et de Transfert des connaissances » a été ajoutée en facteur commun des 4 actions scientifiques afin de répondre à la question fondamentale des échanges entre acteurs.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00199/31006/35663.pdf
http://archimer.ifremer.fr/doc/00199/31006/

Éditeur(s) : Elsevier Sci Ltd
Résumé : Many questions remain unanswered regarding RNAi-based mechanisms and dsRNA-induced antiviral immune responses in penaeid shrimp. In this study, we report the characterization in the white leg shrimp Litopenaeus vannamei of RNAi pathway associated proteins Lv-Ago 1 and Lv-Ago 2, two members of the Argonaute family of proteins, as well as Lv-sid 1, the first shrimp homologue of Sid-1, a membrane channel-forming protein implicated in the cellular import of dsRNA. To decipher their functional implication in RNAi-related phenomena, we monitored their relative expression following stimulation by specific and non-specific RNA duplexes of diverse length. The findings show that the length of small RNA duplexes plays a critical role in the activation of both RNAi-related and innate antiviral responses. They also suggest that these two mechanisms of antiviral response may activate the same pathway, requiring Lv-Sid 1 and Lv-Ago 2 induction. (C) 2010 Elsevier Ltd. All rights reserved.
Developmental And Comparative Immunology (0145-305X) (Elsevier Sci Ltd), 2010-11 , Vol. 34 , N. 11 , P. 1209-1218

Droits : 2010 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/00015/12652/9587.pdf
DOI:10.1016/j.dci.2010.06.017
http://archimer.ifremer.fr/doc/00015/12652/

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) : Wiley-blackwell Publishing, Inc
Résumé : Vibrioses are the predominant bacterial infections in marine shrimp farms. Vibrio nigripulchritudo is an emerging pathogen of the cultured shrimp Litopenaeus stylirostris in New Caledonia and other regions in the Indo-Pacific. The molecular determinants of V. nigripulchritudo pathogenicity are unknown; however, molecular epidemiological studies have revealed that recent pathogenic V. nigripulchritudo isolates from New Caledonia all cluster into a monophyletic clade and contain a small plasmid, pB1067. Here, we report that a large plasmid, pA1066 (247 kb), can also serve as a marker for virulent V. nigripulchritudo, and that an ancestral version of this plasmid was likely acquired prior to other virulence-linked markers. Additionally, we demonstrate that pA1066 is critical for the full virulence of V. nigripulchritudo in several newly developed experimental models of infection. Plasmid pB1067 also contributes to virulence; only strains containing both plasmids induced the highest level of shrimp mortality. Thus, it appears that these plasmids, which are absent from non-pathogenic isolates, may be driving forces, as well as markers, for the emergence of a pathogenic lineage of V. nigripulchritudo.
Environmental Microbiology (1462-2912) (Wiley-blackwell Publishing, Inc), 2011-02 , Vol. 13 , N. 2 , P. 296-306

Droits : 2010 Society for Applied Microbiology and Blackwell Publishing Ltd
http://archimer.ifremer.fr/doc/00031/14196/11469.pdf
DOI:10.1111/j.1462-2920.2010.02329.x
http://archimer.ifremer.fr/doc/00031/14196/

Éditeur(s) : The 11th Pacific Science Inter-Congress : Pacific Countries and their Ocean: Facing Local and Global Changes. March 2 - 6, 2009 Tahiti, French Polynesia
Résumé : New Caledonian shrimp farming feasibility studies started in the mid-seventies. Today ca. 2000 tons of Litopenaeus stylirostris, introduced in 1978, are semi-intensively produced each year. Since fifteen years, production has been impacted by two seasonal vibriosis, the “Summer” syndrome caused by Vibrio nigripulchritudo in relation to pond bottom and water column quality, and the “Winter Syndrome” caused by Vibrio penaeicida in relation to drastic temperature drops . With the ambition of elucidating processes possibly leading to mortality outbreaks in ponds, Ifremer has been conducting a research program since 2003 under the DESANS project (2003-2006) and the present DEDUCTION project, equally funded by the South and North Provinces and by the Government of New Caledonia. This program has been designed in a multidisciplinary approach integrating the suspected multifactorial origin of shrimp mortalities. Environment quality in ponds (either sediment or water column) plays a major role in the shrimp ecophysiological status, influencing sensitivity to pathogens . DEDUCTION focuses on a better knowledge of phytoplankton composition and biogeochemical fluxes at the sediment-water interface in ponds. Impact of farm discharges is also investigated. In addition, a more fundamental work is conducted to understand the pathogenicity mechanisms of these two Vibrio species but also to prevent the occurrence of viral infections through a RNA interference approach. Markers of the ecophysiological status of broodstock, larvae, post-larvae and reared animals, such as enzymes testifying from an oxidative stress, are developed. Gain of performance (resistance to pathogens and growth) were obtained on hybrids of the New Caledonian and introduced Hawaiian strains. Finally the database Stylog is exploited in view of discriminating “good” from “bad” rearings on the basis of relevant indicators.
Les études de faisabilité de l'aquaculture de crevettes ont débuté en Nouvelle-Calédonie dans les années soixante dix. Aujourd'hui environ 2000 tonnes de Litopenaeus stylirostris, introduite en 1978, sont produites chaque année de façon semi-intensive. Depuis quinze ans la production a été impactée par deux vibrioses : le syndrome d'été provoqué par Vibrio nigripulchritudo en relation avec la qualité de la tranche d'eau et des fonds de bassins et le syndrome d'hiver provoqué par Vibrio penaeicida en relation avec des chutes rapides de température. Dans le but d'élucider les processus entrainant des mortalités dans les élevages, Ifremer a conduit un programme de recherche depuis 2003 dans le cadre du projet DESANS (2003-2006) puis du présent projet DEDUCTION co-financé par les provinces Nord et Sud et le Gouvernement de la Nouvelle-Calédonie. Ce programme se construit dans une démarche multidisciplinaire intégrant les origines supposées multifactorielles des mortalités de crevettes. La qualité environnementale dans les bassins (les fonds et la colonne d'eau) joue un rôle majeur dans l'état physiologique des animaux, influençant leur sensibilité aux pathogènes. DEDUCTION se focalise sur une meilleure compréhension de la composition phytoplanctonique et des flux bio-géochimiques à l'interface eau/sédiment dans les bassins.L'impact des rejets des fermes est également étudié. En outre, un travail plus fondamental est conduit pour connaître les mécanismes de pathogénicité de ces deux espèces de Vibrio mais aussi pour prévenir les occurences d'infections virales par des approches ARN Interférence. Des marqueurs de l'état physiologique des oeufs, larves, post-larves et animaux en élevage, tels que les enzymes du stress oxydant sont utilisés.Les gains de performance (résistance aux pathogènes et croissance) sont obtenus sur des hybrides issus du croisement de la souche locale avec une souche hawaienne. Finalement la base de données STYLOG est exploitée dans le but de distinguer les bonnes des mauvaises pratiques sur la base d'indicateurs fiables.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00143/25438/23591.pdf
http://archimer.ifremer.fr/doc/00143/25438/

Éditeur(s) : Elsevier Science Bv
Résumé : In New Caledonia, shrimp hatcheries are confronted with mass mortality in the larval stages, a phenomenon poorly understood as no specific causative agent has been identified. This has resulted in an excessive use of prophylactic antibiotics, although their adverse effects in aquaculture are notorious. The present work was thus aimed at selecting potential probiotic strains for penaeid hatcheries. From a pool of more than 400 marine bacterial isolates sampled from the local marine environment seven strains exhibited in vitro antagonistic activity towards Vibrio harveyi. These isolates were characterized both phenotypically and genotypically using a biochemical approach and 16S rDNA sequencing. Six out of these seven strains were found to belong to the genus Pseudoalteromonas, the last one belonging to the Vibrionaceae family and related to the Harveyi clade. Selected probiotic candidates were individually tested for antagonistic activities in vitro using a green fluorescent protein (GFP)-labelled transconjugant of V. harveyi and for inherent pathogenicity towards cultured shrimp larvae at two different developmental stages. This approach enabled rapid processing and selection of candidates to be tested in our experimental hatchery. Four different experiments were conducted to test candidate strains either alone or in combination. Repeated trials showed that postlarval survival was significantly improved by adding the strain NC201 individually to the rearing water compared to unchallenged controls. The analysis of immune-related gene expressions showed that the Litsty PEN3 transcript abundance of larvae was significantly increased after being reared in probiotic-containing water; however no significant difference in lysozyme gene expression was recorded in this study. Together, these results open new insights into the use of these strains as potential substitutes to antibiotherapy in shrimp larval rearing in New Caledonia.
Aquaculture (0044-8486) (Elsevier Science Bv), 2014-08 , Vol. 432 , P. 475-482

Droits : 2014 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/00190/30129/28591.pdf
DOI:10.1016/j.aquaculture.2014.04.031
http://archimer.ifremer.fr/doc/00190/30129/

Éditeur(s) : Academic Press Ltd- Elsevier Science Ltd
Résumé : Over the last decade, RNA interference pathways have emerged in eukaryotes as critical regulators of many diverse biological functions including, among others, transcriptional gene regulation, post-transcriptional gene silencing, heterochromatin remodelling, suppression of transposon activity, and antiviral defences. Although this gene silencing process has been reported to be relatively well conserved in species of different phyla, there are important discrepancies between plants, invertebrates and mammals. In penaeid shrimp, the existence of an intact and functional RNAi machinery is supported by a rapidly growing body of evidence. However, the extent to which this process participates to the host immune responses remains poorly defined in this non-model organism. This review summarizes our current knowledge of RNAi mechanisms in shrimp and focuses on their implication in antiviral activities and shrimp immune defences.
Fish & Shellfish Immunology (1050-4648) (Academic Press Ltd- Elsevier Science Ltd), 2013-04 , Vol. 34 , N. 4 , P. 1002-1010

Droits : 2012 Elsevier Ltd. Published by Elsevier Ltd
http://archimer.ifremer.fr/doc/00119/23063/20899.pdf
DOI:10.1016/j.fsi.2012.06.008
http://archimer.ifremer.fr/doc/00119/23063/

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) : 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) : 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 : injection of non-specific dsRNA initiates a broad-spectrum innate antiviral immune response in the Pacific white shrimp, Litopenaeus vannamei, however, the receptor involved in recognition of this by-product of viral infections remains unknown. In vertebrates, dsRNA sensing is mediated by a class of Toll-like receptors (TLRs) and results in activation of the interferon system. Because a TLR (lToll) was recently characterized in L. vannamei, we investigated its potential role in dsRNA recognition. We showed that injection of non-specific RNA duplexes did not modify lToll gene expression. A reverse genetic approach was therefore implemented to study its role in vivo. Silencing of lToll did not impair the ability of nonspecific dsRNA to trigger protection from white spot syndrome virus and did not increase the shrimp susceptibility to viral infection, when compared to controls. In contrast, gene-specific dsRNA injected to specifically silence lToll expression activated an antiviral response. These data strongly suggest that shrimp lToll plays no role in dsRNA-induced antiviral immunity. (C) 2009 Elsevier Ltd. All rights reserved.
Developmental & Comparative Immunology (0145-305X) (Elsevier), 2009-07 , Vol. 33 , N. 7 , P. 806-810

Droits : 2009 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/2009/publication-6428.pdf
DOI:10.1016/j.dci.2009.02.005
http://archimer.ifremer.fr/doc/00000/6428/