Effet des conditions environnementales sur le développement des pathologies à Vibrio dans les élevages de crevettes en Nouvelle-Calédonie Auteur(s) : Lemonnier, Hugues 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/ | Partager |
A review of the known hosts geographical range and current diagnostic procedures for the virus diseases of cultured penaeid shrimp Auteur(s) : Lightner, D Bell, T Redman, R Éditeur(s) : Actes de colloques Ifremer, Tahiti, French Polynesia, 20 Feb - 4 Mar 1989, n°9, chap. 13, pp.113-126 Résumé : At least six virus diseases are presently known in cultured penaeid shrimp. Each of these six penaeid virus diseases (BP = Baculovirus penaei ; MBV = Penaeus monodon -type baculovirus (including PBV of Australian penaeids); BMN = baculoviral midgut gland necrosis; HPV = hepatopancreatic parvo-like virus; IHHNV = infectious hypodermal and hematopoietic necrosis virus; and REO = reo-like virus of the hepatopancreas) may be comprised by a multitude of individual strains, some of which are highly pathogenic to some penaeids, while being of little importance to others. Droits : info:eu-repo/semantics/openAccess http://archimer.ifremer.fr/doc/1989/acte-1481.pdf http://archimer.ifremer.fr/doc/00000/1481/ | Partager Voir aussi Baculovirus penaei Penaeus japonicus Penaeus monodon Malacostraca Marine crustaceans Necroses Infectious diseases Viruses Diseases Télécharger |
Aspects moléculaires et biochimiques des stylicines, peptides multifonctionnels identifiés chez la crevette bleue du Pacifique Litopenaeus stylirostris (Crustacea, Decapoda) Auteur(s) : Rolland, Jean-luc Éditeur(s) : Université Montpellier 2 - Sciences et Techniques du Languedoc Résumé : The work reported here was motivated by the economical importance of the pacific blue shrimp Litopenaeus stylirostris farming where high mortality rates are due to bacterial and viral diseases. It consists in the characterisation of two original peptides, the first members of a new multifunctional family of peptides from peneide shrimps, the stylicines. Those two peptides, named stylicines 1 and 2, are negatively charged (pI < 6.0), and characterised by a proline-rich N-terminal region and a C-terminal region containing 13 cysteine residues. Stylicines are synthesized by heamocytes where they are stored within small cytoplasmic granules. To understand the role of these peptides in the immune response of shrimps to a vibrio infection, their recombinant forms were produced in E. coli BL21 (DE3) plysS, purified and characterised. The two rstylicines display biological anti-proliferative and blood clotting activities. Only rstylicine 1 displays antimicrobial activities: antifungal against Fusarium oxysporum (MIC<2.5μM) and bacteriostatic against Gram (−) bacteria, Vibrio sp. (MIC<80μM). Moreover this peptide displays an LPS-binding activity (dissociation constant (Kd) of 9.6×10−8 M) and agglutinate Vibrio. penaeicida "in vitro". Finally, the presence of sequences coding for modified forms of stylicine 1 in some shrimp’s genome may be in relation with their lower ability to survive infections. Les travaux présentés dans ce mémoire ont été motivés par l’importance économique de l’élevage de la crevette bleue du pacifique Litopenaeus stylirostris dont les fortes mortalités sont principalement dues au développement de maladies bactériennes et virales. Ils ont consisté en la caractérisation des deux premiers membres d’une famille originale de peptides multifonctionnels présents chez les crevettes pénéides, les stylicines. Ces peptides, nommés stylicines 1 et 2, sont des peptides anioniques (pI < 6.0), formés d’une région amino-terminale riche en résidus de type proline et d’une région carboxyterminale riche de treize résidus cystéines. Ces molécules sont synthétisées et stockées dans de petits granules présents dans le cytoplasme des hémocytes. Pour mieux appréhender leurs rôles dans la réponse immunitaire des crevettes à une infection par des Vibrio, leurs formes recombinantes ont été produites dans E. coli BL21 (DE3) plysS, purifiées et caractérisées. Les deux rstylicines présentent des activités antiproliférative et anticoagulante. Seule la rstylicine1 présente des activités antimicrobiennes : antifongique sur Fusarium oxysporum (CMI<2.5 μM), et antibactérienne (bactériostatique) sur Vibrio sp (CMI<80 μM). Ce peptide est également capable de se lier aux LPS des bactéries à Gram (-) (Kd= 9.6x10-8 M) et d’agglutiner V. penaeicida "in vitro". Enfin, l’existence de gènes codant des formes modifiées de la stylicine1, chez certaines crevettes, pourrait être en relation avec une diminution de la résistante des individus aux infections. Droits : 2010 The Author, Univ Montpellier 2 http://archimer.ifremer.fr/doc/00024/13558/10612.pdf http://archimer.ifremer.fr/doc/00024/13558/ | Partager |
Nutrient and microbial dynamics in eutrophying shrimp ponds affected or unaffected by vibriosis Auteur(s) : Lemonnier, Hugues Courties, Claude Mugnier, Chantal Torreton, Jean-pascal Herbland, Alain 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/ | Partager |
Biological, physiological, immunological and nutritional assessment of farm-reared Litopenaeus stylirostris shrimp affected or unaffected by vibriosis Auteur(s) : Mugnier, Chantal Justou, Carole Lemonnier, Hugues Patrois, Jacques Ansquer, Dominique Goarant, Cyrille Le Coz, Jean-rene 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/ | Partager |
Probiotic effect of FLOC on Vibrios in the pacific white shrimp Litopenaeus Vannamei Auteur(s) : Aguilera-rivera, Diana Prieto-davo, Alejandra Escalante, Karla Chavez, Cristina Cuzon, Gerard Gaxiola, Gabriela Éditeur(s) : Elsevier Science Bv Résumé : The advantages of FLOC over clear water (CW) in rearing juvenile L. vannamei and its effects on Vibrio communities were evaluated. Survival rate in FLOC and clear water were recorded and a probiotic was tested under both conditions. Daily growth rate (DGR) was higher in FLOC (p < 0.05) than in CW. Survival in each system increased significantly when a probiotic was included in the diet (p < 0.05). The Vibrionaceae community from the hepatopancreas (HP) and the culture medium did not differ between the two culture media. Nevertheless, a novel group of Vibrio strains was found to be unique to FLOC. No high level of lesions was observed in shrimp tissues from the FLOC + probiotic treatment; it suggests that the probiotic contributed to homeostasis and prevented outbreak of opportunistic pathogenic species. Aquaculture (0044-8486) (Elsevier Science Bv), 2014-03 , Vol. 424 , P. 215-219 Droits : 2014 Elsevier B.V. All rights reserved. http://archimer.ifremer.fr/doc/00173/28379/26694.pdf DOI:10.1016/j.aquaculture.2014.01.008 http://archimer.ifremer.fr/doc/00173/28379/ | Partager |
Rapid and sensitive PCR detection of Vibrio penaeicida, the putative etiological agent of syndrome 93 in New Caledonia Auteur(s) : Saulnier, Denis Avarre, Jean-christophe Le Moullac, Gilles Ansquer, Dominique Levy, Peva Vonau, Vincent Éditeur(s) : Inter-Research Résumé : Experimental infections of Penaeus (Litopenaeus) stylirostris were performed with a Vibrio penaeicida strain (AM101) isolated in New Caledonia from Syndrome 93 diseased shrimp. Cumulative mortalities resulting from intramuscular injection or immersion of shrimp in bacterial suspensions demonstrated high virulence for this bacterial strain and suggested that V. penaeicida could be the etiological agent of Syndrome 93. The median lethal dose (LD50) for AM101 was 1.3 x 104 CFU (colony forming units) ml-1 by immersion and less than 5 CFU shrimp-1 by intramuscular challenge, with mortality outbreaks at 48 and 22 h after challenge, respectively. A polymerase chain reaction (PCR) detection assay using a primer set designed from the 16S ribosomal RNA gene of V. penaeicida was developed. It gave an expected amplicon of approximately 310 bp in ethidium bromide-stained agarose gels. The specificity of these primers was assessed with different Vibrio species. Furthermore, DNA extracted by the ChelexTM method could be used to detect fewer than 20 cultured Vibrio cells in seawater or shrimp hemolymph by this assay. It appears to be a reliable screening method for detecting V. penaeicida in shrimp and from the aquatic environment. Diseases of aquatic organisms (0177-5103) (Inter-Research), 2000-03 , Vol. 49 , N. 2 , P. 109-115 Droits : Inter-Research 2000 http://archimer.ifremer.fr/doc/2000/publication-4436.pdf DOI:10.3354/dao040109 http://archimer.ifremer.fr/doc/00000/4436/ | Partager |
Eutrophication in a tropical pond: Understanding the bacterioplankton and phytoplankton dynamics during a vibriosis outbreak using flow cytometric analyses Auteur(s) : Lucas, Ronan Courties, C. Herbland, Alain Goulletquer, Philippe Marteau, Anne-laure Lemonnier, Hugues Éditeur(s) : Elsevier Science Bv Résumé : In tropical shrimp ponds, the increasing of feed input, concomitantly with the stocking shrimp biomass, induces an eutrophication of the ecosystem. Although difficult to maintain, its stability is required to guarantee the success of the culture. A 110-day period of phytoplankton and bacterioplankton stock and dynamics in an earthen pond (1.2 ha area, 1 m depth) was monitored using flow cytometry to provide baseline information on community characteristics and ecosystem instability. Seven autotrophic cell types were identified over the whole sampling period. Prokaryotic cells included Synechococcus sp., a group named UNK which presented an atypical new flow cytometric signature and picoeukaryotes (PEUK). Nanophytoplankton cells were represented by 4 groups: NAN1, NAN2, NAN3 and Cryptophytes. During the first part of the survey, picophytoplankton dominated the phytoplanktonic assemblage. The mean abundance of total cells (up to 8 x 10(6) cells mL(-1)) was among the highest recorded in marine and brackish waters. Bacterial abundance and production ranged from 0.8 to 5.1 x 10(7) cells mL(-1) and from 30 to 110 mu g C L-1 h(-1). A shift from pico to nanophytoplankton abundance was observed for a few days from d 96. During this period, heterotrophic bacteria production and abundance suddenly dropped, implying a change in the functioning of the microbial loop. This shift was concomitant with a significant shrimp mortality outbreak due to Vibrio penaeicida, the etiological agent of a disease known as Syndrome 93, which affects the shrimp industry in New Caledonia. This survey suggests that flow cytometric analysis could be used for the monitoring of aquaculture systems to improve our understanding of the complex phytoplankton and bacterial dynamics of these systems and its potential influence on disease development. (c) 2010 Elsevier B.V. All rights reserved. Aquaculture (0044-8486) (Elsevier Science Bv), 2010-12 , Vol. 310 , N. 1-2 , P. 112-121 Droits : 2010 Elsevier B.V. All rights reserved. http://archimer.ifremer.fr/doc/00029/14068/11312.pdf DOI:10.1016/j.aquaculture.2010.10.022 http://archimer.ifremer.fr/doc/00029/14068/ | Partager Voir aussi Tropical shrimp aquaculture Ecosystem shift Phytoplankton Bacterioplankton Vibrio Flow cytometry Télécharger |
Identification de marqueurs génétiques de la virulence chez Vibrio nigripulchritudo, un pathogène de crevettes pénéides en Nouvelle-Calédonie Auteur(s) : Reynaud, Yann 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/ | Partager Voir aussi plasmide SSH épidémiology virulence shrimp vibriosis Vibrio nigripulchritudo plasmide SSH épidémiologie Télécharger |
Pathogenicity of Vibrio penaeicida or white shrimp Litopenaeus vannamei: a cysteine protease-like exotoxin as a virulence factor Auteur(s) : Aguirre Guzman, G Ascencio, F Saulnier, Denis Éditeur(s) : Inter-Research Résumé : The pathogenicity of Vibrio penaeicida Strains KH-1 and AM101, their culture-free supernatant (CFS), and their protein fraction obtained by 40% of ammonium sulfate precipitation (PFs40) were assessed in experimental challenges against juvenile Litopenaeus vannamei. Live Vibrio cells, CFS, and PFs40 from the AM101 strain produced a significantly higher mortality (p < 0.05) compared to the KH-1 strain. Toxicity and median lethal doses (LD50) of Fast Protein Liquid Chromatography (FPLC) products were evaluated on L. vannamei. The first FPLC fraction sample (A) from PFs40 of the AM101 strain displayed LD50 values of 1.68 and 5.61 mu g protein ind.(-1), respectively. The second FPLC process from Fraction A showed a peak (A1) also with toxic effects to shrimp. PFs40, Fraction A, and Peak At showed a 38.5 kDa molecular band (SDS-PAGE), with activity on a gelatin protease zymogram. The lethal effect of PFs40 and Fraction A was inhibited by Proteinase K, CuCl2, E-64, and heat (60 and 100 degrees C) treatments, but was not inhibited by EDTA-Na-2, aprotinin, and soy trypsin treatments. These results and the zymogram inhibition test suggest the presence of a cysteine protease-like proteinaceous exotoxin as a dominant protease, secreted by V. penaeicida Strain AM101. Diseases of aquatic organisms (0177-5103) (Inter-Research), 2005-11 , Vol. 67 , N. 3 , P. 201-207 Droits : Inter-Research 2005 http://archimer.ifremer.fr/doc/2005/publication-3618.pdf DOI:10.3354/dao067201 http://archimer.ifremer.fr/doc/00000/3618/ | Partager |
Résistance de la crevette Litopenaeus stylirostris à la bactérie pathogène Vibrio penaeicida : Physiologie, immunologie et pathologie comparées d’une population sélectionnée sur un critère de survie aux épisodes de mortalité et d’une population témoin non sélectionnée. Auteur(s) : De Decker, Sophie Résumé : The New-Caledonian shrimp industry is based on the controlled reproduction of the shrimp Litopenaeus stylirostris, a species which was introduced in the 80s. The major difficulty to which the industry has been faced for 10 years is the occurrence of the “Syndrome 93”, which corresponds to mortality phases when the temperature falls down in April-May-June. This mortality is associated to the pathogenic bacteria Vibrio penaecida. An experiment of genetic selection based on the criterion ofsurvival to Syndrome 93 has been conducted at the Laboratory of Aquaculture of New Caledonia. The 3rd selected generation had demonstrated very encouraging results (survival rates improved by around 20% in experimental infections with V. penaeicida). These results have not been confirmed at the 4th generation and no difference in terms of physiology and immunology appears between the selected population and the non-selected control population. The potential causes of these results are examined and proposals for protocol improvements are given. La filière crevette de Nouvelle-Calédonie reposesur la maîtrise de la reproduction contrôlée de la crevette Litopenaeus stylirostris, espèce introduite dans les années 1980. La difficulté majeure que rencontre la filière depuis une dizaine d’années est la récurrence du « Syndrome 93 », qui s’exprime sous forme d’épisodes de mortalités lors des baisses de température aux intersaisons. Ces mortalités sont associées à la bactérie pathogène Vibrio penaeicida Une expérience de sélection sur un critère de survie à des épisodes de Syndrome 93 a été menée au Laboratoire Aquacole de Calédonie. La 3èmegénération sélectionnée avait montré des résultats très encourageants (survies améliorées de l’ordre de 20% lors d’infections expérimentales à V. penaeicida). Ces résultats ne sont pas confirmés en 4ème génération et aucune différence en termes physio-et immunologique n’apparaît entre la population sélectionnée et la population témoin non sélectionnée. Les causes potentielles de ces résultats sont examinées et des propositions d’amélioration de protocoles sont avancées. Droits : 2004 Université de la Rochelle, Ifremer http://archimer.ifremer.fr/doc/00204/31527/29943.pdf http://archimer.ifremer.fr/doc/00204/31527/ | Partager |
Sequence polymorphism-based identification and quantification of Vibrio nigripulchritudo at the species and subspecies level targeting an emerging pathogen for cultured shrimp in New Caledonia Auteur(s) : Goarant, Cyrille Reynaud, Yann Ansquer, Dominique De Decker, Sophie Merien, Fabrice 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/ | Partager |
Quantification of Vibrio penaeicida, the etiological agent of Syndrome 93 in New Caledonian shrimp, by real-time PCR using SYBR Green I chemistry Auteur(s) : Goarant, Cyrille Merien, F 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/ | Partager |
Arbitrarily primed PCR to type Vibrio spp. Pathogenic for shrimp Auteur(s) : Goarant, Cyrille Merien, F Berthe, Franck Mermoud, I Perolat, P Éditeur(s) : Amer Soc Microbiology Résumé : A molecular typing study on Vibrio strains implicated in shrimp disease outbreaks in New Caledonia and Japan was conducted by using AP-PCR (arbitrarily primed PCR). It allowed rapid identification of isolates at the genospecies level and studies of infraspecific population structures of epidemiological interest. Clusters identified within the species Vibrio penaeicida were related to their area of origin, allowing discrimination between Japanese and New Caledonian isolates, as well as between those from two different bays in New Caledonia separated by only 50 km. Other subclusters of New Caledonian V. penaeicida isolates could be identified, but it was not possible to link those differences to accurate epidemiological features. This contribution of AP-PCR to the study of vibriosis in penaeid shrimps demonstrates its high discriminating power and the relevance of the epidemiological information provided. This approach would contribute to better knowledge of the ecology of Vibrio spp. and their implication in shrimp disease in aquaculture. Applied And Environmental Microbiology (0099-2240) (Amer Soc Microbiology), 1999-03 , Vol. 65 , N. 3 , P. 1145-1151 Droits : American society for microbiology http://archimer.ifremer.fr/doc/1999/publication-2734.pdf http://archimer.ifremer.fr/doc/00000/2734/ | Partager |
Les échanges internationaux de crevettes pénéidés vivantes entre les pays du Pacifique, de l'Atlantique et de l'Europe Auteur(s) : Michel, Alain Éditeur(s) : Office Int Epizooties Résumé : The tremendous development of penaeid shrimp culture across the world over the past twenty years has led to international trade in eggs, larvae and spawners of the best shrimp species for aquaculture. Trade has involved, in particular, the following species: - Penaeus japonicus from Japan to Europe, the Pacific Islands and South America - P. monodon from South-East Asia to almost all tropical countries - P. vannamei and P. stylirostris from countries along the Pacific coast of South and Central America to the United States of America, the islands of the South Pacific, countries along the Atlantic coast of the Americas and certain countries in Africa. In the 1980s, research conducted by American and French teams enabled the breeding of stocks of spawners in captivity. This resulted in the establishment of lines of captive broodstock outside natural areas of distribution, and the development of new breeding farms in previously unproductive regions. Trade related to movements of these species is facilitated by the absence of legislation in most countries; even when such legislation does exist, it is not always applied These practices have led to the rapid development of world shrimp production, but have also contributed to the dissemination of pathogens. In recent years, the occurrence of serious epizootics - occasioning heavy losses world-wide - has led to a more cautious approach involving trade of post-larvae obtained from captive broodstock in which thorough control measures have been implemented for known diseases. Trade in wild shrimp will probably be abandoned progressively, to avoid the risk of introducing new pathogens. The main challenges for research in the near future are the development of diagnostic tools, to enable continuous control of captive broodstock, and the selection of strains which are resistant to the principal pathogens affecting these species. Revue Scientifique Et Technique-office International Des Epizooties (0253-1933) (Office Int Epizooties), 1996-06 , Vol. 15 , N. 2 , P. 499-515 Droits : info:eu-repo/semantics/openAccess http://archimer.ifremer.fr/doc/00091/20231/17883.pdf http://archimer.ifremer.fr/doc/00091/20231/ | Partager |
"Summer Syndrome" in Litopenaeus stylirostris in New Caledonia: Pathology and epidemiology of the etiological agent, Vibrio nigripulchritudo Auteur(s) : Goarant, Cyrille Ansquer, Dominique Herlin, Jose Domalain, David Imbert, Frederic De Decker, Sophie 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/ | Partager |
Searching for genetic markers of virulence in Vibrio nigripulchritudo, a Litopenaeus stylirostris shrimp pathogen Auteur(s) : Reynaud, Yann 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/ | Partager |
Emergence de maladies chez les organismes d'intérêt aquacole : quelques scénarios illustrés d'exemples Auteur(s) : Saulnier, Denis Reynaud, Yann Arzul, Isabelle Miossec, Laurence Le Roux, Frédérique Goarant, Cyrille 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/ | Partager Voir aussi Cyprinus carpio Ostrea edulis Bonamia ostreae Peneides Vibrio nigripulchritudo Herpesvirus Agent pathogène Télécharger |
Pathotyping of Vibrio Isolates by Multiplex PCR Reveals a Risk of Virulent Strain Spreading in New Caledonian Shrimp Farms Auteur(s) : Labreuche, Yannick Pallandre, Laurane Ansquer, Dominique Herlin, Jose Wapotro, Billy Le Roux, Frederique 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/ | Partager |
A white spot disease-like syndrome in the Pacific blue shrimp (Litopenaeus stylirostris) as a form of bacterial shell disease Auteur(s) : Goarant, Cyrille Brizard, Raphael Marteau, Anne-laure Éditeur(s) : Elsevier Résumé : In May 1997, some white lesions evoking the white spot syndrome disease were observed in Litopenaeus stylirostris broodstock in New Caledonia. The occurrence of these lesions was neither associated with mortality, nor with histological evidence of white spot syndrome baculovirus (WSBV), The evidence suggests that these lesions result from a form of bacterial disease and are associated with an increased bacterial flora on the outer surface of the cuticle, as well as an increased incidence throughout the molt cycle. A microscopic observation of these lesions allows them to be differentiated from WSBV-associated lesions. Aquaculture (0044-8486) (Elsevier), 2000-03 , Vol. 183 , N. 1-2 , P. 25-30 Droits : 2000 Elsevier Science B.V. All rights reserved http://archimer.ifremer.fr/doc/2000/publication-373.pdf DOI:10.1016/S0044-8486(99)00284-7 http://archimer.ifremer.fr/doc/00000/373/ | Partager Voir aussi New Caledonia white spot syndrome Aquaculture Pathogenic bacteria Shrimp culture Crustaceans Diseases Litopenaeus stylirostris Télécharger |