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/

Éditeur(s) : Université de La Rochelle
Résumé : Shrimp aquaculture in New Caledonia is a developing industry, exclusively based on the species Litopenaeus stylirostris. However, it is subject to mortalities during grow out as observed all over the world. Since 1993, shrimp reared during the cold period are affected by a disease named "syndrome 93". As a consequence, the whole industry has become seasonal and produces essentially during the warm season. The climatic conditions during the periods of transition between the two main seasons could be responsible for an environmental stress and the origin reason for mortality outbreaks. Stocking density is the main risk factor. It increases the probability of the appearance and development of the epizooty in the pond. The "Summer Syndrome", a septicemic vibriosis caused by Vibrio nigripulchritudo, was identified for the first time in December 1997 in an intensive shrimp farm (DF). It has become enzootic ever since and is expressed after 50d of rearing. Since 2003, two others farms located near DF have developed the disease. As a consequence, the summer syndrome is considered to be a potential threat for all the industry. Between 1991 and 2005, the increase of feed input and nitrogen levels in feed has led to a significant increase in the growth rate over the years in the farms affected by the disease. In consequence, an early eutrophication of the water is observed, which is concomitant with the beginning of the mortality outbreaks. An early eutrophication and the virulence level of the pathogen are considered as the main risk factors of the disease. Factors may act synergistically to explain the summer syndrome. As part of a multidisciplinary approach (pathology – physiology – environment), two high-frequency surveys were carried out to analyse the pond ecosystem. Since the feed quantity increases with the biomass of shrimp, the eutrophication level of the pond ecosystem will also tend to increase with the duration of rearing. During the first part of the rearing, the abundance of each picoplankton type is exceptionally high and picophytoplankton dominated the autotrophic compartment. The nanophytoplankton dominated the second part of the rearing and was more unstable. The shift from pico to nanophytoplankton could be considered as an environmental stress and was observed in relation to the beginning of the mortalities, whatever the disease. Shift intensity could play a role by inducing directly or indirectly a stress for shrimp and/or a growth and/or virulence factors of the pathogen. Results from sediment studies show that mortality was best characterized by TAN concentration in pore water and even more when the TAN concentration was high. TAN and pH are potential stress factors for shrimp reared in ponds affected by these diseases.
L'aquaculture mondiale de crevettes est un secteur à très forte croissance. Toutefois, les problèmes liés aux maladies dans les élevages ne cessent d'augmenter depuis 1980. Au cours du développement de la filière de Nouvelle-Calédonie, deux épisodes saisonniers de mortalités associées à la présence de Vibrio pathogènes sont apparus et ont fortement perturbé les techniques et les schémas de production. Les premières mortalités hivernales, décrites sous le nom de « syndrome 93 » ont été observées en mai 1993. Elles ont affecté depuis tous les élevages de saison froide de toutes les fermes de production. Les conditions climatiques pendant les périodes de transitions entre les deux principales saisons caractérisant le climat de la Nouvelle-Calédonie, seraient responsables d'un stress environnemental à l'origine du déclenchement du syndrome 93. La densité initiale d'ensemencement, apparaît comme le principal facteur de risque, augmentant la probabilité d'apparition de la maladie et l'ampleur de ses effets. Le « syndrome d'été » a été identifié pour la première fois en 1997 dans une ferme pratiquant une production intensive. Il s'est déclaré depuis 2003 dans deux autres fermes géographiquement proches. Les mortalités apparaissent 58 jours en moyenne après l'ensemencement des post-larves dans les bassins. La recherche d'une croissance maximale des animaux a induit, avec les années, une intensification progressive du système caractérisée par une augmentation de la quantité d'aliment distribuée. L'examen des données historiques fait apparaître une eutrophisation du milieu de plus en plus précoce concomitante au déclenchement des mortalités. En l'état actuel de nos connaissances, cette évolution précoce de l'écosystème en présence de façon récurrente de la souche hautement pathogène du Vibrio incriminé et à une période où la fréquence des mues des crevettes est élevée, constitue le principal facteur de risque de la maladie. Avec l'apport croissant en aliment, la colonne d'eau évolue vers une autotrophie croissante alors que le sédiment devient de plus en plus hétérotrophe au fur et à mesure que l'élevage progresse. Au cours du processus d'eutrophisation, le compartiment autotrophe montre une succession de deux assemblages. Le picophytoplancton domine sur la première partie de l'élevage et laisse place à du nanophytoplancton en seconde partie. Cette phase de transition, concomitante à l'apparition des mortalités quel que soit le syndrome, caractérise un stress environnemental dont l'intensité pourrait favoriser ou non le déclenchement des mortalités. Les suivis de différents indicateurs dans les sédiments - pH, potentiel d'oxydoréduction et concentration en ammoniaque dans l'eau interstitielle – indiquent dans les bassins déclarant les épizooties des conditions qui ont été définies expérimentalement comme potentiellement plus stressantes pour les animaux.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2007/these-2643.pdf
http://archimer.ifremer.fr/doc/00000/2643/

Éditeur(s) : Elsevier
Résumé : A field survey was conducted on two intensive shrimp farms using similar technical practices: one (DF) historically affected by a vibriosis, the other (HC) in which the pathogen has been observed although no mortality event has occurred. Because historical data suggest that eutrophication process may directly or indirectly play a role in the disease outbreak, we focussed our research on its dynamics. A higher variability of the phytoplanktonic compartment linked to an imbalance in the molar N:P ratio was observed in farm DF compared to farm HC, implying a modification on the linkage between the bacteria and phytoplankton compartments at DF. The beginning of the mortality outbreak at DF followed a shift from picoto nanophytoplankton. The organic matter mineralization process at the water-sediment interface may explain the disturbance observed in the water column during eutrophication. The consequences of this disturbance on shrimps' health status and pathogen ecology are discussed. (C) 2009 Elsevier Ltd. All rights reserved.
Marine Pollution Bulletin (0025-326X) (Elsevier), 2010-03 , Vol. 60 , N. 3 , P. 402-411

Droits : 2010 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/00002/11368/8119.pdf
DOI:10.1016/j.marpolbul.2009.10.012
http://archimer.ifremer.fr/doc/00002/11368/

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) : 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) : Université de Paris 6 – Pierre et Marie Curie
Résumé : Since 1997, a new pathology seasonally occurs in new caledonian shrimp farms during the warm season and was named Summer Syndrome. Diseased Litopenaeus stylirostris shrimp suffer from a septicemic vibriosis which was attributed to V. nigripulchritudo. Preliminary studies based on a collection of V. nigripulchritudo strains have brought to light different virulence levels according to experimental infections results; three virulence statuses were defined: highly (HP), moderately (MP) and non pathogenic (NP). The aim of this work was to genetically characterize virulent V. nigripulchritudo strains. In a first step the genetic diversity of 58 V. nigripulchritudo strains was analyzed by MLST and AP-PCR, revealing a cluster of HP and MP strains, characterized by a low genetic variability and that includes all Summer Syndrome-associated isolates. This confirms the emergence of one cluster of pathogenic V. nigripulchritudo simultaneously with the emergence of the Summer Syndrome ; in a second step, 368 genetic markers of virulence were identified by a Suppressive Subtractive Hybridization performed between the genomes of a HP strain and a genetically close, NP isolate; the distribution of the screened SSH fragments was studied in 58 V. nigripulchritudo isolates by macro-array: 78 DNA fragments were selected, allowing to characterize clusters identified and pathogenic statuses; 13 are specific of the HP strains involved in Summer Syndrome. Interestingly, 10 of these markers are carried by a plasmid pSFn1 that contains sequences highly similar to those of a plasmid pAK1, detected in Vibrio shilonii, a coral pathogen. The origin and consequences of this plasmid acquisition are discussed.
Depuis 1997, les élevages de crevettes en Nouvelle-Calédonie sont confrontés à une nouvelle maladie, le Syndrome d'été, une vibriose septicémique dont l'agent étiologique est Vibrio nigripulchritudo. Les résultats d'infection expérimentale sur une collection de souches, ont montré l'existence de trois pathotypes distincts : hautement (HP), moyennement (MP) et non pathogène (NP). L'étude du polymorphisme génétique de 58 souches par typage moléculaire en MLST et AP-PCR, a mis en évidence un groupe phylogénétique particulier caractérisé par un très faible degré de variabilité génétique (confirmant l'émergence de ce groupe en parallèle à l'émergence du Syndrome d'été) et constitué uniquement de souches HP (dont toutes celles associées au Syndromed'été) et de souches MP. Afin d'identifier des marqueurs génétiques de la virulence des souches responsables du Syndrome d'été, et parmi ces marqueurs des gènes codant potentiellement pour des effecteurs de la virulence, une approche soustractive par SSH a été développée entre une souche HP de type Syndrome d'été et une souche NP : 368 marqueurs génétiques ont ainsi été mis en évidence ; la distribution de ces marqueurs a été étudiée chez les 58 souches de la collection par une approche en macroarray : 78 marqueurs ont été sélectionnés, qui permettent de caractériser les différents groupes phylogénétiques et les différents pathotypes, dont 13 fragments spécifiques des souches HP type Syndrome d'été. Parmi ces 13 fragments, 10 ont été localisés sur le plasmide pSFn1 qui a été entièrement séquencé. Ce même plasmide a été purifié uniquement des souches HP de type Syndrome d'été. Par ailleurs, une très forte homologie a été mise en évidence entre pSFn1 et pAK1, un autre plasmide également séquencé et retrouvé chez la souche V. shilonii AK1, responsable du blanchiment du corail Oculina patagonica en Méditerranée. Ces résultats ont ouvert la discussion sur le rôle de pSFn1 dans la virulence de V. nigripulchritudo.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2008/these-3906.pdf
http://archimer.ifremer.fr/doc/00000/3906/

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) : Agro Paris Tech
Résumé : P. acidilactici MA 18/5M is a dietary probiotic (BACTOCELL®) which has shown its efficiency on several terrestrial and aquatic species. The present thesis aimed to investigate more fully the effects of this probiotic on cultured penaeid shrimps. For this study, we chose a global approach by replacing the problem studied in a model of interactions between three compartments: the shrimp, its intestinal microflora and the probiotic. Having first defined how the probiotic should be administrated, we studied its effects under controlled laboratory conditions at several levels of Litopenaeus stylirostris biology: (i) Nutritional status and growth, (ii) associated bacterial microflora, and (iii) health status with respect to oxidative stress. Then the probiotic improves the growth of the shrimps and the feed conversion ratio. Our study shows that these effects are partially due to a better use of the feed, particularly of dietary carbohydrates, but also suggest an action on the metabolism, andor on the growth via a contribution of essential nutrients. On the other hand, although the intestinal bacterial communities seems little affected, we show an antagonism effect of the probiotic toward total and vibrionacea intestinal flora, as well as a decrease in the level of infection of shrimps during challenge with pathogenic V. nigripulchritudo. Beside, our works highlight that infection by this pathogen, led to a reduction in antioxidant defences, to a rise in oxidative damages, and to a release of mortalities within 48 hours. This oxidative stress following bacterial infection was better controlled by animals receiving the probiotic diet, and resistance of the shrimps to infection was significant improved. A nutritional hypothesis is proposed to explain such result, hypothesis based on a link between carbohydrate metabolism and antioxidant status. As a conclusion of these works, we can propose a first explanatory scenario for the effects of P. acidilactici on a penaeid shrimp. Finally probiotic's effects were assessed under a pilot scale in a commercial farm from New Caledonia currently affected by the "summer syndrome ". We confirm the improvements brought in term of zootechnical results, with in particular, a better resistance to this syndrome. This work was made possible thanks to a breeding method in floating cages, developed during this thesis, and therefore transferred, as an experimental tool, to the shrimp industry of New Caledonia.
Pediococcus acidilacticii MA18/5M est un probiotique bactérien (BACTOCELL®) qui a démontré son efficacité chez plusieurs espèces d'animaux terrestres et aquatiques. Cette thèse avait pour objectif d'investiguer plus en détails les effets de ce probiotique chez les crevettes pénéides d'élevage. Pour cette étude, nous avons choisi une approche globale replaçant la problématique dans un modèle d'interactions entre trois compartiments : la crevette, sa microflore intestinale et le probiotique. Après avoir défini les modalités d'administration, l'effet de P. acicilactici a été étudié, en conditions de laboratoire, chez la crevette Litopenaeus stylirostris, à différents niveaux: (i) statut nutritionnel et croissance, (ii) microflore bactérienne associée, et (iii) santé de l'animal à travers son statut antioxydant et son niveau de stress oxydant. Ainsi, le probiotique améliore la croissance des crevettes et l'indice de conversion de l'aliment. Notre étude montre que ces effets sont en partie liés à une meilleure utilisation de l'aliment, notamment des carbohydrates alimentaires, mais suggèrent aussi une action sur le métabolisme, et/ou sur la croissance via un apport de nutriments essentiels. D'autre part, bien que les communautés bactériennes intestinales semblent peu affectées, un effet d'antagonisme du probiotique vis-à-vis de la flore intestinale totale et vibrionacée a été montré, ainsi qu'une réduction du niveau d'infection des crevettes au cours de challenge avec le pathogène V. nigripulchritudo. Par ailleurs, nous mettons en évidence que l'infection par ce pathogène, conduit à un abaissement des défenses antioxydantes, à une élévation des dégâts oxydatifs, et au déclenchement de mortalités dans les 48 heures. Le probiotique atténue le niveau de ce stress oxydant, et améliore significativement la résistance des crevettes à l'infection. Une hypothèse nutritionnelle est proposée afin d'expliquer ce résultat, hypothèse reposant sur un lien entre le métabolisme des carbohydrates et le statut antioxydant. A l'issue de ces travaux, nous pouvons alors proposer un premier scénario explicatif des effets de P. acidilactici chez une crevette pénéide. Finalement le probiotique a été éprouvé à l'échelle pilote sur une ferme commerciale de Nouvelle-Calédonie affectée par le « syndrome d'été ». Nous confirmons les améliorations apportées en termes de résultats zootechniques, avec notamment une meilleure résistance au syndrome. Ce travail a été réalisé grâce une méthode d'élevage en cages flottantes, mis au point au cours de cette thèse, et qui a dés lors été transférée, comme outil expérimental, à la filière crevetticole Calédonienne.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2009/these-6814.pdf
http://archimer.ifremer.fr/doc/00000/6814/

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 : The study of the antioxidant status and oxidative stress in shrimp L. stylirostris make it possible to assess risk factors (temperature, shrimp size, pathogen, ..) associated with shrimp farming. From a practical view, this research should lead to zootechnical and/or nutritional recommendations to improve the antioxidant status of the shrimp during their growout stage in farm.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00197/30875/29243.pdf
http://archimer.ifremer.fr/doc/00197/30875/

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/

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) : 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 Latin American shrimp Litopenaeus stylirostris was introduced in three different Pacific islands (Tahiti, New Caledonia via Tahiti, and Hawaii) and hatchery-propagated for 7-25 generations to develop shrimp farming based on these domesticated stocks. Three microsatellite markers have been used in an attempt to assess the genetic bases of the populations available to start a selective breeding program. The comparison of eight hatchery stocks (five New Caledonian, two Hawaiian and one Tahitian stocks) and one wild Ecuadorian population showed a much lower variability in the domesticated stocks than in the wild population, especially in New Caledonia and Tahiti (2-3.7 vs. 14-27 alleles per locus; 20-60% vs. 90% expected heterozygosity). The Tahitian and the New Caledonian stocks share the same alleles, suggesting that the loss of alleles occurred during the common past of these populations. On the contrary, New Caledonian and Hawaiian populations share only one common allele at the three loci studied. Although the low genetic variability and the resulting inbreeding of the New Caledonian stocks do not seem to affect their present performance, the results of this study demonstrate the usefulness of the introduction of new stocks in order to increase the potential responses to new controlled or uncontrolled selective pressures. The introduction in New Caledonia of the Hawaiian domesticated stocks, which would provide the local shrimp industry with 40% of the allelic diversity of the species, is advised and preferred to the one of wild animals in order to take advantage (i) of the spontaneous selection which occurred during domestication and (ii) of their favourable sanitary "specific pathogen free" status (no presence of four viruses: WSV, YHV, IHHNV, TSV) which limits the risk of introduction of pathogens.
La crevette d'Amérique latine Litopaenus stylirostris a été introduite dans trois îles du Pacifique (à Tahiti, en Nouvelle-Calédonie via Tahiti, et à Hawaii), et a été ensuite reproduite en écloserie pendant 7 à 25 générations à des fins d'aquaculture. Trois marqueurs microsatellites ont été utilisés pour évaluer les bases génétiques des populations disponibles pour le démarrage d'un programme d'amélioration génétique. L'étude comparative de 8 populations domestiquées (cinq néo-calédoniennes, deux hawaiiennes et une tahitienne) et d'une population sauvage d'Equateur révèle une variabilité très réduite dans les populations d'élevage, en particulier en Nouvelle-Calédonie et à Tahiti (2 à 3.7 allèles par locus au lieu de 14 à 27 en population sauvage ; 20% à 60% d'hétérozygotie au lieu de 90%). Les souches tahitiennes et calédoniennes disposent des mêmes allèles, ce qui suggère que la perte d'allèles a eu lieu lors de l'histoire commune des ces populations. A l'inverse, les populations néo-calédoniennes et hawaiiennes n'ont en commun qu'un seul allèle sur les 3 locus étudiés. Bien que la très faible variabilité génétique du cheptel calédonien ne semble pas affecter ses performances actuelles, les résultats de cette étude démontrent l'utilité de l'introduction de variabilité afin d'augmenter la capacité de réponse à de nouvelles pressions de sélection (contrôlées ou non). L'introduction des souches hawaiiennes en Nouvelle-Calédonie qui permettrait à la filière locale de disposer de 40% de la diversité allélique de l'espèce) est préconisée de préférence à celle d'animaux sauvages afin de bénéficier (i) de la sélection spontanée qu'elles ont subi lors de leur domestication et (ii) de leur statut sanitaire « specific pathogen free, SPF » (absence de 4 virus : WSV, YHV, IHHNV, TSV) qui limite les risques de transferts de pathogènes.
Aquatic Living Resources (0990-7440) (Elsevier), 2003-12 , Vol. 16 , N. 6 , P. 501-508

Droits : 2003 Published by Elsevier, Paris.
http://archimer.ifremer.fr/doc/2003/publication-596.pdf
DOI:10.1016/j.aquliv.2003.07.001
http://archimer.ifremer.fr/doc/00000/596/

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/

Résumé : Shrimp new blood importation under strict biosecurity conditions requires multiple measures so as to ensure that pathogens, unidentified during the pre-import tests, might be introduced and propagated to the shrimp farms. Closed cycle quarantine facilities using recycled artificial seawater are an adequate answer to this challenge by allowing secure rearings and additional pathogen testing.
L’importation de sang neuf dans les meilleures conditions de biosécurité impose que de multiples précautions soient prises afin d’éviter que la présence d’un pathogène, non identifié lors des tests préliminaires à l’importation, puisse se propager à l’ensemble de la filière. L’utilisation d’installations de quarantaine fonctionnant en circuit fermé permet de répondre à ce souci en réalisant l’élevage des crevettes importées, sur une période de plusieurs mois, dans des conditions d’isolement tout en permettant la réalisation de tests de détection de pathogènes.

Droits : 2004 Ifremer
http://archimer.ifremer.fr/doc/00117/22811/20615.pdf
http://archimer.ifremer.fr/doc/00117/22811/

Résumé : Penaeus stylirostris shrimp imported from Hawaii for a genetic program and kept at the Saint-Vincent research facility had progressively developed a high sensitivity to IHHNV, the etiological agent of the Infectious, Hypodermic, and Hematopoietic Necrosis. Fears were that the virus could not only be transferred to their offspring, but also that the New Caledonian strain of Penaeus stylirostris, so far considered as resistant, could become sensitive by an off balance of its hosts/pathogen equilibrium. In October 2008, a joint decision between Uprac (shimp owner), Davar (New Caledonian Animal Health Authority) and Ifremer (scientific partner) was taken to terminate all pure breed and hybrid Hawaiian populations remaining in the ponds, and to implement an intensive fallowing protocol, and controlled restocking. IHHNV was also suspected to trouble experiments carried out in the research facility (mortalities, deformities, slow growth), thus affecting research programs. Difficulties pointed out by IHHNV have triggered since 2008 some awareness on biosecurity issues at some level of the New Caledonian shrimp industry. At the St-Vincent research station, it led to a risk assessment and implementation of biosecurity procedures (fallowing, propagation, grow out and use of screened animals) which minimizes the risk of expression of IHHNV. The use of a brand new hatchery building on site also helped in resetting routine hygiene procedures during production and in controlling incoming and outgoing items (shrimp, water, feed, personnel, equipment, pests and wastes). IHHNV has not been detected on analysed samples since then nor have any symptoms been found during experimental or broodstock growout. Keeping this control alive is considered as a useful tool to lower the risk of potential health threats, to get a better understanding in case of an outbreak, and to increase the reliability of experimental results.
Les crevettes originaires d’Hawaii et conservées au LEAD St-Vincent, dans le cadre du programme d’introduction de variabilité génétique dans le cheptel Calédonien, ont progressivement montré une sensibilité importante au virus IHHNV, agent de la Nécrose Hypodermale et Hématopoïétique Infectieuse (mortalité, croissance ralentie, déformations). Les risques d’une transmission génétique de cette sensibilité à leur descendance à moyen terme, et de son développement par rupture de l’équilibre hôte/pathogène sur la souche dite « Calédonienne » jusqu’alors considérée comme résistante à l’IHHN, ont conduit à partir d’octobre 2008, avec l’accord et l’appui de l’UPRAC et de la DAVAR, à la suppression des animaux hawaiiens ou hybrides encore présents dans les bassins de terre du LEAD St-Vincent, au vide sanitaire de ces derniers, et au repeuplement contrôlé du site. D’autre part, le virus IHHN est soupçonné d’avoir pu affecter les animaux utilisés dans les expérimentations menées au LEAD-St-Vincent (mortalités, déformations, croissance réduite), influençant donc potentiellement leur bon déroulement et celui des programmes de recherches. Les obstacles soulevés par l’expression du virus IHHN ont donc initié depuis 2008 une prise de conscience et la mise en place de démarches de biosécurité dans certains points de la filière. A la station de St-Vincent, cette démarche s’est traduite par une identification/évaluation des risques et l’application de mesures (vide sanitaire, reproduction, élevage et utilisation des animaux contrôlés) chacune dotée de procédures qui regroupées, minimisent le risque d’introduction et de dissémination de l’IHHNV. La prise en main de la nouvelle structure d’écloserie sur le site de la SASV a également été mise à profit pour revoir les procédures d’hygiène de routine dans la conduite des productions et dans le contrôle des flux intrants et sortants (animaux, eau, aliments, personnel, équipement, nuisibles, déchets). Aucune détection d’IHHNV sur les échantillons prélevés n’a depuis été observée, de même qu’aucun symptôme de l’expression de cette maladie sur les animaux en élevage sur le site de Saint-Vincent. La poursuite de ces mesures est envisagée comme un outil d’aide à la minimisation des problèmes potentiels, à leur compréhension en cas d’apparition, et à la fiabilisation des résultats.

Droits : 2010 Ifremer
http://archimer.ifremer.fr/doc/00118/22949/20776.pdf
http://archimer.ifremer.fr/doc/00118/22949/

Éditeur(s) : Elsevier
Résumé : The "Summer Syndrome", a septicemic vibriosis caused by Vibrio nigripulchritudo, is responsible for one of the two main seasonal mortalities which affect shrimp aquaculture in New Caledonia. It was identified for the first time in December 1997 in an intensive shrimp farm (called farm DF) and has been enzootic ever since. The "Summer syndrome" affects shrimp during warm-season growout. Although the geographic area concerned is limited, it is a potential threat for the industry in that the disease could spread. Analysis of grow out data from 1991 to 2002 in this farm was carried out in order to characterize the disease and the environmental conditions in relation to mortality. Results of farm DF were compared to data from farms not affected by the disease (called farms HC). The "Summer syndrome" occurs during the warm season which is characterized by an increase in water temperature and sunny duration. This disease is characterized by simultaneously highest densities and weakest drying duration between crops in the farms studied. Between 1991 and 2002, the increase of feed input and nitrogen rate in feed has led to a significant increase of the growth rate over the years in farm DF. At the same time, water management has not changed and has induced an increase in environmental waste production. In consequence, an early eutrophication of water is observed, which is concomitant with the beginning of the mortality outbreaks. This could play a role by inducing directly or indirectly a stress for shrimp and/or a growth and/or virulence factors of the pathogen.
Aquaculture (0044-8486) (Elsevier), 2006-08 , Vol. 261 , N. 3 , P. 1039-1047

Droits : 2006 Elsevier
http://archimer.ifremer.fr/doc/2006/publication-1868.pdf
DOI:10.1016/j.aquaculture.2006.08.036
http://archimer.ifremer.fr/doc/00000/1868/

Éditeur(s) : Elsevier
Résumé : Vibriosis is a major disease problem in shrimp aquaculture, affecting all developmental stages, from larvae in hatchery tanks to juveniles and broodstock in growout ponds. However, bacterial strains responsible for vibriosis in the successive stages are usually considered to be different, and virulence specificity has been reported both at the species and at the stage levels. The so-called « Syndrome 93 » is a seasonal juvenile vibriosis caused by Vibrio penaeicida which affects Penaeus stylirostris in growout ponds and broodstock tanks in New Caledonia. This pathology does not cause any mortality in hatchery or nursery phases. An experimental infection design using balneation of postlarvae and early juveniles in V. penaeicida suspensions was used to evaluate the developmental stage at which shrimp become sensitive to this vibriosis. We demonstrated that the acquisition of susceptibility to this pathogen is very sudden and correlated with the acquisition of the definitive rostral formula, and from this draw conclusions regarding virulence mechanisms of V. penaeicida in P. stylirostris.
Aquaculture (Elsevier), 1998-12 , Vol. 169 , N. 3-4 , P. 291-296

Droits : 1998 Elsevier Science B.V. All rights reserved
http://archimer.ifremer.fr/doc/1998/publication-383.pdf
DOI:10.1016/S0044-8486(98)00380-9
http://archimer.ifremer.fr/doc/00000/383/

Éditeur(s) : American society for microbiology
Résumé : Vibrio nigripulchritudo, the etiological agent of Litopenaeus stylirostris summer syndrome, is responsible for mass mortalities of shrimp in New Caledonia. Epidemiological studies led to the suggestion that this disease is caused by an emergent group of pathogenic strains. Genomic subtractive hybridization was carried out between two isolates exhibiting low and high virulence. Our subtraction library was constituted of 521 specific fragments; 55 of these were detected in all virulent isolates from our collection (n = 32), and 13 were detected only in the isolates demonstrating the highest pathogenicity (n = 19), suggesting that they could be used as genetic markers for high virulence capacity. Interestingly, 10 of these markers are carried by a replicon of 11.2 kbp that contains sequences highly similar to those of a plasmid detected in Vibrio shilonii, a coral pathogen. The detection of this plasmid was correlated with the highest pathogenicity status of the isolates from our collection. The origin and consequence of this plasmid acquisition are discussed.
Applied and environmental microbiology (depuis 1953) (0099-2240) (American society for microbiology), 2008-05 , Vol. 74 , N. 10 , P. 3038-3047

Droits : 2008 American Society for Microbiology
http://archimer.ifremer.fr/doc/2008/publication-4523.pdf
DOI:10.1128/AEM.02680-07
http://archimer.ifremer.fr/doc/00000/4523/

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/