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 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) : Academic Press Inc Elsevier Science
Résumé : Several Vibrio species are known to be pathogenic to the Pacific oyster Crassostrea gigas. Survival varies according to pathogen exposure and high mortality events usually occur in summer during gametogenesis. In order to study the effects of gametogenetic status and ploidy (a factor known to affect reproduction allocation in oysters) on vibriosis survival, we conducted two successive experiments. Our results demonstrate that a common bath challenge with pathogenic Vibrio splendidus and Vibrio aestuarianus on a mixture of mature, spawning and non-mature oysters can lead to significant mortality. Previous bath challenges, which were done using only non-mature oysters, had not produced mortality. Immunohisto-chemical analyses showed the affinity of Vibrio for gonadic tissues, highlighting the importance of sexual maturity for vibriosis infection processes in oysters. Mortality rate results showed poor repeatability between tanks, however, in this bath challenge. We then tested a standardized and repeatable injection protocol using two different doses of the same combination of two Vibrio species on related diploid and triploid oysters at four different times over a year. Statistical analyses of mortality kinetics over a 6-day period after injection revealed that active gametogenesis periods correspond to higher susceptibility to vibriosis and that there is a significant interaction of this seasonal effect with ploidy. However, no significant advantage of triploidy was observed. Triploid oysters even showed lower survival than diploid counterparts in winter. Results are discussed in relation to differing energy allocation patterns between diploid and triploid Pacific oysters. (C) 2010 Elsevier Inc. All rights reserved.
Journal Of Invertebrate Pathology (0022-2011) (Academic Press Inc Elsevier Science), 2011-02 , Vol. 106 , N. 2 , P. 179-191

Droits : 2010 Elsevier Inc. All rights reserved.
http://archimer.ifremer.fr/doc/00030/14108/11368.pdf
DOI:10.1016/j.jip.2010.09.003
http://archimer.ifremer.fr/doc/00030/14108/

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

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

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

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

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

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

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

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

Éditeur(s) : 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) : Academic Press Ltd- Elsevier Science Ltd
Résumé : The understanding of reciprocal interactions between Crassostrea gigas and Vibrio sp., whether these be virulent or avirulent, is vital for the development of methods to improve the health status of cultured oysters. We describe an original non-invasive experimental infection technique using cohabitation, designed to explore these interactions. Using real-time PCR techniques we examined the dynamics of virulent and avirulent Vibrio sp. in oyster hemolymph and tank seawater, and made a parallel study of the expression of four genes involved in oyster immune defense: Cg-BPI, Cg-EcSOD, Cg-IκB, Cg-TIMP. No mortality occurred in control animals, but oysters put in cohabitation for 2–48 h with animals previously infected by two Vibrio pathogens suffered mortalities from 2 to 16 days post-cohabitation. Our results show that virulent Vibrio infect healthy individuals after only 2 h of cohabitation, with values ranging from 4.5×102 to 2×104 cells ml−1 hemolymph. Simultaneously, an approximate ten-fold increase of the total Vibrio population was observed in control animals, with a 6.6–78.5-fold up-expression of targeted genes. In contrast, oysters exposed to harmful bacteria had mean expression levels strongly down-regulated by a factor of 9.2–29 (depending on the gene) compared with control animals. Although oysters were still found to be infected by virulent Vibrio after 6–48 h of cohabitation, no significant differences were noted when comparing levels of each transcript in control and infected oysters at the same sampling times during this period: the important differences were noted before 6 h cohabitation. Taken together, our data support (1) the hypothesis that virulent Vibrio disturbs the immune response of this invertebrate host both rapidly and significantly, although this occurs specifically during an early and transient period during the first 6 h of cohabitation challenge, and that (2) expression of targeted genes is not correlated with vibriosis resistance.
Fish & Shellfish Immunology (1050-4648) (Academic Press Ltd- Elsevier Science Ltd), 2011-02 , Vol. 30 , N. 2 , P. 691-699

Droits : 2011 Elsevier Ltd All rights reserved.
http://archimer.ifremer.fr/doc/00025/13604/10681.pdf
DOI:10.1016/j.fsi.2010.12.017
http://archimer.ifremer.fr/doc/00025/13604/

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

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

Éditeur(s) : Elsevier
Résumé : Shrimp farming is a small but growing industry in New Caledonia. Since 1993, "Syndrome 93" has been affecting New Caledonian shrimp farming industry every cold season, causing severe epizootic mortalities in grow-out ponds and significant losses. Highly pathogenic strains of Vibrio penaeicida are considered the etiological agent of the disease in Litopenaeus stylirostris. On one hand, studies demonstrated that healthy shrimp may carry V penaeicida for weeks with a high overall prevalence, regardless of any seasonal pattern or temperature conditions. On the other hand, larvae are free of V penaeicida and are also resistant to experimental infection. V penaeicida is frequently detected in incoming water pumped from the bays, which was shown, by a molecular typing study, to be the infectious source. This particular epidemiological pattern highlights the major role of the factors that trigger and aggravate the disease in grow-out ponds, where shrimp populations carry the pathogen all year round. In order to gain a better understanding of "Syndrome 93" epidemiology, quantification of V penaeicida both in shrimp and the shrimp farm ecosystem is necessary. This article describes the steps in the successful development of a real-time PCR quantification assay of V penaeicida in shrimp haemolymph, seawater (from ponds or bays) and sediment pore water, including the choice of an accurate extraction technique. The entire detection method; including sample processing, DNA extraction and real-time PCR amplification, can be completed within 4 h. (c) 2006 Elsevier B.V All rights reserved.
Journal of Microbiological Methods (0167-7012) (Elsevier), 2006-10 , Vol. 67 , N. 1 , P. 27-35

Droits : 2006 Elsevier
http://archimer.ifremer.fr/doc/2006/publication-1903.pdf
DOI:10.1016/j.mimet.2006.02.013
http://archimer.ifremer.fr/doc/00000/1903/

Éditeur(s) : Elsevier
Résumé : in this study we evaluated the effects of the probiotic Pediococcus acidilactici MA18/5M on shrimp, Litopenaeus stylirostris (also called Penaeus stylirostris), first on antioxidant defences and secondly on the oxidative stress status in the shrimps' haemolymph and digestive gland. We conducted two experiments with the same protocol in which shrimps were fed two diets for three weeks: a control diet and a probiotic diet containing 1 g of live P. acidilactici MA18/5M kg(-1). In the first experiment, the shrimps were found to be healthy over the trial period; no mortalities and non-detectable signs of infection were recorded. These resulted in high final survival rates (above 90% in both treatments). On the other hand, during the second trial, carried out at a period of increased risk for an outbreak of the summer syndrome, higher mortalities were recorded, associated with high V. nigripulchritudo prevalence and loads in the shrimp haemolymph. In healthy shrimps (trial 1), no detectable response of the antioxidant defence system and of oxidative stress bio-indicators were observed. However, feeding the probiotic significantly increased the total antioxidant status (TAS) and glutathione peroxidase activity, while all other parameters remained significantly unchanged. In the shrimps exposed to V nigripulchritudo (trial 2), the antioxidant response was characterized by higher antioxidant enzyme activities (superoxide dismutase and catalase) and higher oxidative stress level in the digestive gland (higher oxidized/reduced glutathione ratio, higher malonclialdehyde and carbonyl protein contents) compared to levels found in trial 1. However, shrimps fed the probiotic diet exhibited (i) significantly higher final survival rates (67 +/- 3%) compared to the control (47 +/- 4%) and (ii) a lower prevalence of V nigripulchritudo throughout the trial. Moreover, the antioxidant response and the oxidative stress level recorded in the digestive gland with shrimps submitted to the probiotic diet were lower. In the light of these results, we confirm that P acidilactici enrichment in shrimps' diet seems an effective way of reducing the susceptibility of shrimps to bacterial pathogens. (c) 2009 Elsevier B.V. All rights reserved.
Aquaculture (0044-8486) (Elsevier), 2009-09 , Vol. 294 , N. 3-4 , P. 306-313

Droits : 2009 Elsevier B.V. All rights reserved.
http://archimer.ifremer.fr/doc/2009/publication-6780.pdf
DOI:10.1016/j.aquaculture.2009.06.016
http://archimer.ifremer.fr/doc/00000/6780/