Résumé : A new strain of L. stylirostris was introduced in New Caledonia. Quarantine facilities were set up inland using RAS and artificial seawater. After 5 months of rearing, all the 16 initial families were still represented with a 50% average survival. Three samplings for known pathogens were made during that period, all of them negative. Half the animals were taken to outdoor ponds for rearing to reproduction size when the rest was used to test different arrangements for a biosecure rearing until reproduction. Numerous spawnings and nauplii were obtained but larval rearings could not be completed. Different options are considered for the set up of a biosecure facility allowing the rearing and the breeding of pathogen free strains of shrimp.
Afin de disposer d’une plus forte diversité génétique exploitable, des producteurs calédoniens ont, en relation avec l’Ifremer, récemment introduit d’Hawaii une autre souche de crevette de l’espèce Litopenaeus stylirostris, domestiquée et garantie exempte de pathogènes. Malgré cette garantie sanitaire, les crevettes ont été maintenues en observation dans une quarantaine tertiaire pendant cinq mois avant la sortie et l’élevage d’une moitié de l’effectif en bassins terre extérieurs. L’autre moitié a été conservée dans les installations de quarantaine comme stock de secours au cas où un problème affecterait les crevettes élevées à l’extérieur. Les installations de quarantaine ont été progressivement transformées, tout en continuant les élevages, afin de réaliser un prototype de conservatoire biosécurisé. Les crevettes ont été élevées jusqu’à la taille de géniteurs en utilisant de l’eau de mer artificielle puis de l’eau de mer naturelle traitée au chlore. Des essais de reproduction et d’élevage larvaire ont été menés et de nombreuses pontes fécondées ont été obtenues mais les élevages larvaires, à une exception, n’ont pas dépassé le stade Zoé 3. Les analyses par PCR réalisées sur les crevettes du conservatoire ont montré que les mesures et précautions sanitaires qui avaient été prises avaient préservé le statut sanitaire “sans pathogènes” initial. Cette expérience a permis de mieux cerner les problèmes qui pouvaient se poser pour la mise en place et le fonctionnement d’installations biosécurisées utilisant des circuits fermés. Ces enseignements ont servi à répertorier les principales contraintes envisageables pour la réalisation d’un futur conservatoire crevette qui viendrait conforter et sécuriser les souches actuellement disponibles en Nouvelle-Calédonie. Plusieurs options sont proposées pour les crevettes devant entrer dans le conservatoire (sans quarantaine, quarantaine tertiaire, quarantaine primaire puis secondaire) et pour le type de fonctionnement du conservatoire. Les principaux critères pour le choix du site, l’utilisation de l’eau de mer et son traitement, les systèmes de recirculation sont abordés; et des schémas d’installations de quarantaine et de conservatoire sont proposés.

Droits : 2007 Ifremer
http://archimer.ifremer.fr/doc/00117/22849/20659.pdf
http://archimer.ifremer.fr/doc/00117/22849/

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) : 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/

Éditeur(s) : Elsevier
Résumé : Two populations of the Latin American shrimp Penaeus (Litopenaeus)stylirostris domesticated in Hawaii and in New Caledonia were previously shown to be genetically differentiated and proven highly inbred. In New Caledonia, where different Vibriosis affect shrimp production and antibiotic use is banned in growing ponds, the Hawaiian population was introduced to increase the allelic variability available for local shrimp farmers and start a genetic improvement program. Growth and survival rates of the two pure populations and the two-way F-1-hybrids obtained by breeding Hawaiian animals with New Caledonian animals were assessed in several simple experiments (earthen ponds, floating cages and experimental infection challenges) during two years on two successive generations. Results were very consistent: F-1-hybrids growth rates in earthen ponds were 37% (+/-7% SD) higher than for pure populations. Cage experiments demonstrated no competition between the different populations when reared together or separately in a common environment. The F-1-hybrids also showed better survival rates in all experiments. Combining the results on growth and survival rates leads to the conclusion that biomass production is much higher with F-1-hybrid populations than with pure populations using the same quantity of juveniles stocked: biomass production in ponds was increased 1.4 and 2.3 times on year I and year 2 respectively, and 1.9 times in floating cages. The advantage of growing F-1-hybrids appeared proportionally higher when environmental and sanitary conditions led to poorer survival (34% in year 2 vs. 56% in year 1). These results are a good example of performance improvement by heterosis effect and/or of performance loss due to inbreeding in the pure populations. This study demonstrates that aquaculture industries which cannot afford large selection programs may benefit from using two different inbred parental stocks to produce F-1-hybrids for each commercial growout. This is notably true when only inbred populations are available, or when introduction of genetic variability from the wild or from other genetic resources represents a zoo-sanitary risk. In our case, the expected increase in L stylirostris production could be around 85% (according to our average results) if producers keep stocking their ponds at their current densities using F-1-hybrids. However, for sustainability reasons, it is advisable to stock F-1-hybrid animals at lower densities, the gain in performance allowing producing the same amount of biomass with less input. (C) 2008 Elsevier B.V. All rights reserved.
Aquaculture (0044-8486) (Elsevier), 2008-06 , Vol. 278 , N. 1-4 , P. 43-50

Droits : 2008 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/2008/publication-4316.pdf
DOI:10.1016/j.aquaculture.2008.03.018
http://archimer.ifremer.fr/doc/00000/4316/

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 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/

Résumé : Selective breeding of shrimp for viral resistance is established for species like L. vannamei and L. stylirostris. However, selection for bacterial infection resistance is not documented. In New Caledonia, "Syndrome 93" is a vibriosis that affects cultured shrimps during the cool season with survival from 15 to 30%. Current practises for breeders rearing at low densities do not improve the resistance of the offsprings mainly because selection pressures for "Syndrome 93" are low. Even after the fifth generation, survival improvement is not significant.
La sélection génétique de lignées de crevettes résistantes à certains virus a démontré son efficacité en particulier chez L. vannamei et L. stylirostris. En revanche, la sélection de crevettes résistantes à une bactérie du genre Vibrio est peu documentée. En Nouvelle-Calédonie, le « Syndrome 93 », vibriose à V. penaeicida, affecte toutes les fermes de crevettes L. stylirostris lors des élevages de saison fraîche. Les survies en bassins de grossissement, de l’ordre de 40 à 60% dans un élevage sain, tombent à des valeurs de 15 à 30%. L’élevage de géniteurs à faible densité, s’il maximise les performances de reproduction, ne semble pas optimiser la qualité de la descendance du fait des faibles pressions de sélection qui s’exercent vis-à-vis du syndrome 93. Les améliorations significatives du nombre d’animaux survivant aux infections et aux épisodes de mortalité en bassin restent faibles en valeur absolue à la 5ème génération de sélection expérimentale en conditions d’élevage, mais sur le long terme la stratégie testée pourrait se révéler économiquement efficace.

Droits : 2005 Ifremer
http://archimer.ifremer.fr/doc/00117/22848/20658.pdf
http://archimer.ifremer.fr/doc/00117/22848/

Éditeur(s) : Inter-Research
Résumé : Experimental infections of Penaeus (Litopenaeus) stylirostris were performed with a Vibrio penaeicida strain (AM101) isolated in New Caledonia from Syndrome 93 diseased shrimp. Cumulative mortalities resulting from intramuscular injection or immersion of shrimp in bacterial suspensions demonstrated high virulence for this bacterial strain and suggested that V. penaeicida could be the etiological agent of Syndrome 93. The median lethal dose (LD50) for AM101 was 1.3 x 104 CFU (colony forming units) ml-1 by immersion and less than 5 CFU shrimp-1 by intramuscular challenge, with mortality outbreaks at 48 and 22 h after challenge, respectively. A polymerase chain reaction (PCR) detection assay using a primer set designed from the 16S ribosomal RNA gene of V. penaeicida was developed. It gave an expected amplicon of approximately 310 bp in ethidium bromide-stained agarose gels. The specificity of these primers was assessed with different Vibrio species. Furthermore, DNA extracted by the ChelexTM method could be used to detect fewer than 20 cultured Vibrio cells in seawater or shrimp hemolymph by this assay. It appears to be a reliable screening method for detecting V. penaeicida in shrimp and from the aquatic environment.
Diseases of aquatic organisms (0177-5103) (Inter-Research), 2000-03 , Vol. 49 , N. 2 , P. 109-115

Droits : Inter-Research 2000
http://archimer.ifremer.fr/doc/2000/publication-4436.pdf
DOI:10.3354/dao040109
http://archimer.ifremer.fr/doc/00000/4436/

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/

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) : 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) : 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) : Wiley-blackwell Publishing, Inc
Résumé : Aims: A description of bacterial pathogens in shrimp ponds is necessary to understand their pathogenesis. Vibrio nigripulchritudo was shown to contain saprophytic and pathogenic strains among New Caledonian isolates. We established a method to map the development of V. nigripulchritudo in pond sediments at three different genetic levels: the species level, then at the pathogenic cluster level and finally at the plasmid level, present only in all highly pathogenic isolates. Methods and Results: PCR methods were applied to shrimp pond sediments both before and after a mortality outbreak. Using crude samples, the species V. nigripulchritudo is not detected at first (0/42 samples at day 56 post stocking) but appears frequently in the sediments after the mortality event (30/42 at day 107). The distribution of strains from the pathogenic cluster of V. nigripulchritudo also follows this pattern. In contrast, the pSFn1 virulence-associated plasmid was detected in one sample at day 56 and none at day 107. An enrichment method was developed to lower the detection limits of our assays. After enrichment, the species V. nigripulchritudo was detected in all samples at both dates. The number of samples positive for pSFn1 was 42/42 samples at day 56 and 29/42 at day 107. Conclusions: These results show that the sediments contain V. nigripulchritudo, notably pathogenic strains. Surprisingly, the virulence-associated plasmid pSFn1 found in all V. nigripulchritudo isolated from moribund shrimp appears less frequently in sediments, possibly being useless or even detrimental to its recipient bacteria in this environment. Significance and Impact of the Study: This study confirms the presence of pathogenic V. nigripulchritudo strains in shrimp pond sediment before a mortality outbreak complying with a previous hypothesis that sediments could be the infecting reservoir. After the outbreak, both total V. nigripulchritudo and pathogenic strains populations have largely increased, possibly contributing to the recurrent mortality observed in this shrimp vibriosis.
Journal Of Applied Microbiology (1364-5072) (Wiley-blackwell Publishing, Inc), 2010-06 , Vol. 108 , N. 6 , P. 2003-2011

Droits : 2010 The Society for Applied Microbiology - Wiley-Blackwell
http://archimer.ifremer.fr/doc/00004/11484/8177.pdf
DOI:10.1111/j.1365-2672.2009.04601.x
http://archimer.ifremer.fr/doc/00004/11484/

Éditeur(s) : Elsevier Science Bv
Résumé : Shrimp aquaculture in New Caledonia is subject to seasonal mortalities during grow-out due to highly virulent Vibrio nigripulchritudo (Vn). To understand the mechanisms affecting shrimp resistance and leading to significant mortality, a shrimp ecophysiology and immunology survey was conducted on two farms, the first considered as a "control" farm (HC), the second affected by the disease (DF). Mortality observed during the survey at DF started 50 days after stocking and was typical of this disease. The main observations regarding shrimp were: (a) growth was not affected by the disease and was faster in the DF than in the HC pond; (b) disease did not affect one sex more than the other, or a specific part of the population in terms of weight; (c) the physical condition of shrimp did not specifically allow us to foresee disease outbreak; (d) shrimp at late premolt stage D-2 and early postmolt stage A appeared to be at some points of the mortalities - but not continuously - the most sensitive to disease; (e) physiological, immunological and nutritional parameters of uninfected shrimp in the DF pond were altered, suggesting that environmental stress occurred just before the first mortalities; (f) data suggest that Vn-infected shrimp are more stressed than the presumed healthy shrimp. Combined with pathological and environmental knowledge gained in parallel during this survey, a conceptual model is proposed. Results suggest that an unstable environment induced conditions (i) stressful for the shrimp, increasing their susceptibility to bacterial infections and (ii) favoring the proliferation of the pathogen in the pond. The combination of these two processes could lead to significant mortality.
Aquaculture (0044-8486) (Elsevier Science Bv), 2013-04 , Vol. 388 , P. 105-114

Droits : 2013 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/00176/28764/27567.pdf
DOI:10.1016/j.aquaculture.2013.01.010
http://archimer.ifremer.fr/doc/00176/28764/

Éditeur(s) : Elsevier
Résumé : A collection of 57 isolates of Vibrio nigripulchritudo from either diseased or healthy shrimp and from shrimp farms environment was studied in order to gain a better understanding of the epidemiology of this pathogen, notably isolated from two distinct shrimp disease complexes. Molecular typing using two different techniques, arbitrarily primed PCR (AP-PCR) and multi-locus sequence typing (MLST), studied together with experimental pathology data allowed a relevant epidemiological insight into this possibly emerging pathogen. Additionally, results obtained with the two molecular typing techniques were congruent and allowed discriminating the strains associated with the "Summer Syndrome" from strains isolated from other contexts, especially the other shrimp vibriosis "Syndrome 93". These results highlight that the "Summer Syndrome" is most probably caused by an emergent clonal pathogen that therefore deserves surveillance and that AP-PCR can satisfactorily be used for that purpose
Systematic and Applied Microbiology (0723-2020) (Elsevier), 2006-11 , Vol. 29 , N. 7 , P. 570-580

Droits : 2005 Elsevier
http://archimer.ifremer.fr/doc/2006/publication-1913.pdf
DOI:10.1016/j.syapm.2005.12.005
http://archimer.ifremer.fr/doc/00000/1913/

Éditeur(s) : Elsevier
Résumé : The Summer Syndrome is a new shrimp disease that has been affecting a shrimp growout farm in New Caledonia since end of 1997. It was recognized to be caused by a systemic vibriosis due to Vibrio nigripulchritudo. This new disease turned out almost immediately enzootic in the shrimp farm involved and has affected all its crops ever since. Since the year 2000, V. nigripulchritudo strains have been found in several shrimp farms, although Summer Syndrome is still limited to one particular area, affecting, since 2003, two adjoining farms. As part of a multidisciplinary research program, a high-frequency survey was carried out during the summer 2002-2003 in two shrimp farms: one affected farm and another one in which pathogenic strains could be isolated but without any disease event. It permitted a good description of V. nigripulchritudo dynamics in shrimp and the ecosystem in both farms. The study of virulence characteristics of some isolates showed that pathogenic and non-pathogenic strains occur in the shrimp farms environment and that both may be found at the same time in one farm. Our results strongly suggest a persistence of pathogenic strains in wet zones of the pond sediment at low concentrations between crops in the affected farm, and their development during the rearing cycle as a cause of shrimp infection.
Aquaculture (0044-8486) (Elsevier), 2006-03 , Vol. 253 , N. 1-4 , P. 105-113

Droits : 2005 Elsevier B.V. All rights reserved
http://archimer.ifremer.fr/doc/2006/publication-1110.pdf
DOI:10.1016/j.aquaculture.2005.07.031
http://archimer.ifremer.fr/doc/00000/1110/