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

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

Éditeur(s) : 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) : HAL CCSD Elsevier Masson
Résumé : International audience
The diagnosis of diabetic foot infections is difficult due to limitations of conventional culture-based techniques. The objective of this study was to evaluate the contribution of denaturing gradient gel electrophoresis (DGGE) in the microbiological diagnosis of diabetic foot ulcers in comparison to conventional techniques, and also to evaluate the need to perform a biopsy sample for this diagnosis. Twenty diabetic patients (types 1 and 2) with foot ulcers (grades 1-4) were included. After debridement of their wounds, samples were taken in duplicate by surface swabbing and deep-tissue biopsy. The samples were analyzed by conventional culture and by a new molecular biology tool, DGGE technology. Polymerase chain reaction (PCR)-DGGE led to the identification of more bacteria than did conventional cultures (mean: 2.35 vs 0.80, respectively). In 11 cases, the technology detected pathogenic species not isolated by classical cultures. PCR-DGGE also identified significantly more pathogenic species at deep levels compared with species detected at superficial levels (87% vs 58%, respectively; P = 0.03). In 9/20 cases, pathogenic bacteria were detected only in deep samples, revealing the need to perform tissue biopsy sampling. DGGE, achievable in 48h, could be a useful technique for the bacteriological diagnosis of diabetic foot infections. It may help to identify pathogenic bacteria in deeply infected ulcers, thereby contributing to a more appropriate use of antibiotics.
ISSN: 1262-3636

hal-01122240
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01122240
DOI : 10.1016/j.diabet.2014.03.002
PUBMED : 24751989

Éditeur(s) : HAL CCSD MDPI
Résumé : International audience
Microbial pollution in aquatic environments is one of the crucial issues with regard to the sanitary state of water bodies used for drinking water supply, recreational activities and harvesting seafood due to a potential contamination by pathogenic bacteria, protozoa or viruses. To address this risk, microbial contamination monitoring is usually assessed by turbidity measurements performed at drinking water plants. Some recent studies have shown significant correlations of microbial contamination with the risk of endemic gastroenteresis. However the relevance of turbidimetry may be limited since the presence of colloids in water creates interferences with the nephelometric response. Thus there is a need for a more relevant, simple and fast indicator for microbial contamination detection in water, especially in the perspective of climate change with the increase of heavy rainfall events. This review focuses on the one hand on sources, fate and behavior of microorganisms in water and factors influencing pathogens' presence, transportation and mobilization, and on the second hand, on the existing optical methods used for monitoring microbiological risks. Finally, this paper proposes new ways of research.
ISSN: 1660-4601

hal-01122209
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01122209
PUBMED : 24747537
DOI : 10.3390/ijerph110404292
PUBMEDCENTRAL : PMC4025003

Éditeur(s) : PSIC 11 - 11th Pacific Science Inter-Congress : Pacific Countries and their Ocean: Facing Local and Global Changes / 2nd Symposium on French Research in the Pacific. March 2 - 6, 2009 Tahiti, French Polynesia.
Résumé : Experimental infections together with epidemiological studies have shown that pathogenic and non‐pathogenic isolates of V. nigripulchritudo co‐existed in shrimp farm environment (2, 3). Moreover, obtained results also demonstrated that the “summer syndrome”was caused by a single, possibly emerging, cluster of virulent strains. Consequently, it was hypothesized that pathogenic strains of V. nigripulchritudo may persist from one year to the next in the shrimp farm environment and re‐develop inside the grow‐out system at the following rearing cycle (3). This study was therefore aimed at determining whether V. nigripulchritudo isolates may survive, or not, in a shrimp pond bottom soil during a 18‐week drying period. To this end, V.nigripulchritudo mapping was performed with recently developed molecular tools and classical culture‐dependent techniques.

Droits : info:eu-repo/semantics/openAccess
http://archimer.ifremer.fr/doc/00197/30874/29242.pdf
http://archimer.ifremer.fr/doc/00197/30874/

Éditeur(s) : HAL CCSD BMJ Publishing Group
Résumé : International audience
BACKGROUND: Mutations in the leucine-rich-repeat kinase 2 (LRRK2) gene have been identified in families with autosomal dominant Parkinson's disease (ADPD), the most common of which is the p.G2019S substitution that has been found at varying frequencies worldwide. Because of the size of the LRRK2 gene, few studies have analysed the entire gene in large series of ADPD families. METHODS: We performed extensive mutation analyses of all 51 coding exons of the LRRK2 gene in index cases from 226 Parkinson's disease families compatible with autosomal dominant inheritance, mostly from France (n = 182) and North Africa (n = 14). RESULTS: We found 79 sequence variants, 29 of which were novel. Eight potentially or proven pathogenic mutations were found in 22 probands (9.7%). There were four novel amino acid substitutions that are potentially pathogenic (p.S52F, p.N363S, p.I810V, p.R1325Q) and two novel variants, p.H1216R and p.T1410M, that are probably not causative. The common p.G2019S mutation was identified in 13 probands (5.8%) including six from North Africa (43%). The known heterozygous p.R1441H and p.I1371V mutations were found in two probands each, and the p.E334K variant was identified in one single patient. Most potentially or proven pathogenic mutations were located in the functional domains of the Lrrk2 protein. CONCLUSION: This study leads us to conclude that LRRK2 mutations are a common cause of autosomal dominant Parkinson's disease in Europe and North Africa.
ISSN: 0022-2593

inserm-00529272
http://www.hal.inserm.fr/inserm-00529272
DOI : 10.1136/jmg.2008.062612
PUBMED : 19357115

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/

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
In mutualisms, each interacting species obtains resources from its partner that it would obtain less efficiently if alone, and so derives a net fitness benefit. In exchange for shelter (domatia) and food, mutualistic plant-ants protect their host myrmecophytes from herbivores, encroaching vines and fungal pathogens. Although selective filters enable myrmecophytes to host those ant species most favorable to their fitness, some insects can by-pass these filters, exploiting the rewards supplied whilst providing nothing in return. This is the case in French Guiana for Cecropia obtusa (Cecropiaceae) as Pseudocabima guianalis caterpillars (Lepidoptera, Pyralidae) can colonize saplings before the installation of their mutualistic Azteca ants. The caterpillars shelter in the domatia and feed on food bodies (FBs) whose production increases as a result. They delay colonization by ants by weaving a silk shield above the youngest trichilium, where the FBs are produced, blocking access to them. This probable temporal priority effect also allows female moths to lay new eggs on trees that already shelter caterpillars, and so to occupy the niche longer and exploit Cecropia resources before colonization by ants. However, once incipient ant colonies are able to develop, they prevent further colonization by the caterpillars. Although no higher herbivory rates were noted, these caterpillars are ineffective in protecting their host trees from a pathogenic fungus, Fusarium moniliforme (Deuteromycetes), that develops on the trichilium in the absence of mutualistic ants. Therefore, the Cecropia treelets can be parasitized by two often overlooked species: the caterpillars that shelter in the domatia and feed on FBs, delaying colonization by mutualistic ants, and the fungal pathogen that develops on old trichilia. The cost of greater FB production plus the presence of the pathogenic fungus likely affect tree growth.
PLoS ONE

hal-00912399
https://hal.archives-ouvertes.fr/hal-00912399
https://hal.archives-ouvertes.fr/hal-00912399/document
DOI : 10.1371/journal.pone.0020538
OATAO : 10156

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

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

Éditeur(s) : 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) : HAL CCSD
Résumé : International audience
BACKGROUND AND AIM: The impact of basal core promoter (BCP) and precore (PC) mutants of the hepatitis B virus (HBV) on liver disease severity remains controversial. The aim of the present study was to screen BCP and PC mutations in 252 HBV surface antigen (HBsAg) positive carriers in France and to assess relationships between these mutations and severe fibrosis. METHODS: Direct sequencing of the precore/core gene was used to detect A1762T/G1764A and G1757A mutations in the BCP and G1896A and G1899A mutations in the PC region. RESULTS: The prevalences of A1762T/G1764A, G1757A, G1896A, and G1899A mutations were 34.1%, 38.7%, 54.9%, and 29.3% (P < 0.001), respectively. The independent predictors of severe fibrosis (>/=F3 Metavir) were older age (P < 0.001), male gender (P = 0.012), elevated alanine aminotransferase (P < 0.001), and the double A1762T/G1764A mutant with no other mutations (P = 0.011). Interestingly, the association of the G1899A mutation with the double A1762T/G1764A mutant significantly counteracted the deleterious effect of the sole double A1762T/G1764A mutant (odds ratio [OR] = 0.28 vs. OR = 3.55, respectively, P = 0.028). CONCLUSIONS: Patients with the A1762T/G1764A mutation have a higher risk of severe fibrosis. The G1899A mutation is a protective factor against severe fibrosis that counteracted the deleterious effect of the A1762T/G1764A mutation. Finally, host phenotypic and HBV genotypic markers independently predict fibrosis severity.
Journal of gastroenterology and hepatology

hal-01477209
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477209
DOI : 10.1111/jgh.13338
PUBMED : 26992056

Éditeur(s) : HAL CCSD Kasetsart University
Résumé : http://www.ceropath.org/references/rodent_protocols_book
Technical book
"Protocols for field and laboratory rodent studies" present the best practices for the studies of rodents and rodent-borne pathogens and parasites from the field to the laboratory. It aims at covering the different steps of rodent studies: 1) Trapping, 2) Dissection and preparation of tissue samples for pathogens screening, 3) Identification of species, 4) Isolation of helminths, 5) Preparing rodent specimens for collections. This book gathers accurate recommendations and techniques, based on those generally accepted in the literature and adapted from the experience of the different authors during rodent surveys and laboratory work. Its presentation is designed to work in the fields with a clear and colorful organization of each chapter, with: inserts providing definitions and recommendations, protocols detailed step by step, and an emphasis on illustrations with several photographs. This book is a publication generated by the CERoPath project (Community Ecology of Rodents and their Pathogens in a changing environment, www.ceropath.org) funded by the French Ministry of Research from 2008 to 2011.
http://hal.ird.fr/ird-00714514

ird-00714514
http://hal.ird.fr/ird-00714514
http://hal.ird.fr/ird-00714514/document
IRD : fdi:010056183

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) : HAL CCSD Society for Molecular Biology and Evolution
Résumé : International audience
Vibrio metoecus is the closest relative of Vibrio cholerae, the causative agent of the potent diarrheal disease cholera. Although the pathogenic potential of this new species is yet to be studied in depth, it has been co-isolated with V. cholerae in coastal waters and found in clinical specimens in the USA. We used these two organisms to investigate the genetic interaction between closely related species in their natural environment. The genomes of twenty V. cholerae and four V. metoecus strains isolated from a brackish coastal pond on the US east coast, as well as four clinical V. metoecus strains were sequenced and compared to reference strains. Whole genome comparison shows 86-87% average nucleotide identity (ANI) in their core genes between the two species. On the other hand, the chromosomal integron, which occupies approximately 3% of their genomes, shows higher conservation in ANI between species than any other region of their genomes. The ANI of 93-94% observed in this region is not significantly greater within than between species, meaning that it does not follow species boundaries. V. metoecus does not encode toxigenic V. cholerae major virulence factors, the cholera toxin and toxin co-regulated pilus. However, some of the pathogenicity islands found in pandemic V. cholerae were either present in the common ancestor it shares with V. metoecus, or acquired by clinical and environmental V. metoecus in partial fragments. The virulence factors of V. cholerae are therefore both more ancient and more widespread than previously believed. There is high interspecies recombination in the core genome, which has been detected in 24% of the single-copy core genes, including genes involved in pathogenicity. V. metoecus was six times more often the recipient of DNA from V. cholerae as it was the donor, indicating a strong bias in the direction of gene transfer in the environment.
ISSN: 1759-6653

Droits : http://creativecommons.org/licenses/by/
hal-01264332
http://hal.upmc.fr/hal-01264332
http://hal.upmc.fr/hal-01264332/document
http://hal.upmc.fr/hal-01264332/file/Genome%20Biol%20Evol-2015-Orata-gbe_evv193.pdf
DOI : 10.1093/gbe/evv193

Éditeur(s) : HAL CCSD Public Library of Science
Résumé : International audience
Diagnosis of toxoplasmic encephalitis (TE) in patients with AIDS is not straightforward because clinicians rely initially on an empiric diagnosis based on clinical and radiographic improvement to specific anti-Toxoplasma gondii therapy. There is therefore a need for biological tools to improve the diagnosis of TE, especially in tropical areas where this diagnosis is likely to be underestimated. The use of PCR testing in blood samples for the diagnosis of TE has been limited by its poor sensitivity in the studies conducted in Europe. In tropical South America, the results of PCR sensitivity in blood samples were controversial. Considering that T. gondii strains from tropical South America have substantial genetic and pathogenic differences with those from USA and Europe, it is therefore important to re-evaluate the performance of the PCR assay in blood samples for the diagnosis of TE in AIDS patients from this region. Our results showed that the only interest of PCR would be in the most severe forms of TE with altered consciousness because PCR is more likely to be positive. We also provided important genotyping data on T. gondii strains isolated in human cases of toxoplasmosis in the Caribbean and in South America.
ISSN: 1935-2727

hal-01503761
https://hal-unilim.archives-ouvertes.fr/hal-01503761
https://hal-unilim.archives-ouvertes.fr/hal-01503761/document
https://hal-unilim.archives-ouvertes.fr/hal-01503761/file/journal.pntd.0004790.pdf
DOI : 10.1371/journal.pntd.0004790

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) : HAL CCSD
Résumé : Staphylococcus aureus is an opportunistic pathogen of humans and warm-blooded animals. It is foundin healthy carriage and can cause a wide range of infections in its hosts. The epithelial surface is theprincipal site of staphylococcal infection. The pathogens have evolved sophisticated mechanisms tohijack the main function of the host cells, thus promoting their invasion and colonization. Amongstthese effects are, alteration of host cell apoptosis, promotion of cell proliferation or conversely,inhibition of cell growth. Using human cervix cancer HeLa and mammary gland bovine MAC-Tepithelial cells, we studied the cytopathic effect of S. aureus and its ability to affect host cell cycle.Using microscopic examination, we found that S. aureus induced the cytopathic effect resulting in theenlargement of the host cells 72 hours post infection. Our experiments showed that the proliferationrate of the epithelial cells was slowed down in S. aureus-infected culture. Three different S. aureusstrains from human and animal origins were used and showed similar effect on both human and bovinecells. Microscopic analysis of DAPI-stained cells revealed huge decrease of mitotic indexes in infectedHeLa and Mac-T cells, synchronized by Double Thymidine Block. Using flow cytometry analysis, wefound that S. aureus induces G2/M phase delay in synchronous HeLa cells. By Western blot analysis,we showed that the G2/M phase delay was associated with accumulation of inactive phosphorylatedcyclin-dependent kinase Cdk1/cdc2, a key inducer of mitosis entry, and dephosphorylation of corehistone H3, an important component of the epigenetic landscape.These results suggest that the delay in G2/M phase transition of the infected epithelial cells may beone of the mechanisms employed by S. aureus to survive and to propagate inside the host. A fullcomprehension of the mechanisms associated to S. aureus-host cell interactions will undoubtedly helpto discover novel targets to treat staphylococcal infections.
EMBO
Villars sur Ollon, Switzerland

hal-01454214
https://hal.archives-ouvertes.fr/hal-01454214
PRODINRA : 171657

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/

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) : 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/