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

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

Éditeur(s) : 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) : 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) : 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/

Résumé : Ce travail décrit la mise au point d'un milieu gélosé semi-sélectif visant à mettre en évidence les Vibrio penaecida en élevage larvaire de pénéides. Le ZoBell-Mannitol-Saccharose-Bleu de bromothymol (ZMSB) a un pH élevé, une forte teneur en sels et ne possède que deux sucres acidifiables, le mannitol et le saccharose. Ce milieu peut être utilisé pour la sélection d'un grand nombre d'espèces du genre Vibrio. La salinité et le ph séparent le germe cherché d'une fraction importante d'espèces de Vibrio, et l'acidification du mannitol et du saccharose révèlent (par le changement de couleur du milieu) les Vibrio de type penaecida qui présentent cette caractéristique. De plus, un milieu minimum comme le ZoBell, additionné de glycérol, permet d'augmenter considérablement la production de pigments noirs (grisâtres) insolubles qui s'accumulent sous forme cristalline chez certaines colonies bactériennes comme Vibrio nigripulchritudo, facilitant ainsi la distinction d'espèces physiologiquement proches comme V. penaecida. Les différents aspects des souches testées sont alors observés par repiquages à l'aide de tampons et permettent d'obtenir des images comparables par superposition. Sur les 26 souches testées une seule présente un aspect comparable à V. penaecida sur ZMSB. Ce Vibrio (V. nigripulchritudo) est facilement différenciable de V. penaecida par culture en milieu ZoBell additionné de glycérol. On peut donc conclure à une bonne spécificité du milieu en question et du procédé de repiquage utilisé. En cas de doutes, la présence de V. penaecida est confirmées par quelques tests biochimiques simples. Un protocoles de fabrication d'un sérum anti V. penaecida est également testé.

Droits : 1997 The author, Univ. Caen, Ifremer
http://archimer.ifremer.fr/doc/00032/14340/11624.pdf
http://archimer.ifremer.fr/doc/00032/14340/

Éditeur(s) : Inter-research
Résumé : Vibriosis is a major disease problem in shrimp aquaculture. 'Syndrome 93' is a seasonal juvenile vibriosis caused by Vibrio penaeicida which affects Litopenaeus stylirostris in grow-out ponds in New Caledonia. This study assessed the toxic activities of extracellular products (ECPs) from V: penaeicida, V. alginolyticus and V, nigripulchritudo using in vivo injections in healthy juvenile L. stylirostris (= Penaeus stylirostris) and in vitro assays on shrimp primary cell cultures and the fish cell line epithelioma papulosum cyprini (EPC). Toxic effects of ECPs were demonstrated for all pathogenic Vibrio strains tested both in vivo and in vitro, but for shrimp only; no effect was observed on the fish cell line. ECP toxicity for New Caledonian V. penaeicida was found only after cultivation at low temperature (20 degrees C) and not at higher temperature (30 degrees C). This points to the fact that 'Syndrome 93' episodes are triggered by temperature drops. The assays used here demonstrate the usefulness of primary shrimp cell cultures to study virulence mechanisms of shrimp pathogenic bacteria.
Diseases Of Aquatic Organisms (0177-5103) (Inter-research), 2000-03 , Vol. 40 , N. 2 , P. 101-107

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00000/10543/7600.pdf
DOI:10.3354/dao040101
http://archimer.ifremer.fr/doc/00000/10543/

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

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

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

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

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

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

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

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

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

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

Éditeur(s) : Elsevier
Résumé : 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/

Éditeur(s) : Vibrio 2007, Meeting Pasteur Institute

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/2007/acte-3432.pdf
http://archimer.ifremer.fr/doc/00000/3432/

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) : 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/