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 |
Microsatellite-associated heterosis in hatchery-propagated stocks of the shrimp Penaeus stylirostris Auteur(s) : Bierne, Nicolas Bezuart, Ivan Vonau, Vincent Bonhomme, François Bedier, Edouard Éditeur(s) : Elsevier Résumé : Correlation between DNA microsatellite heterozygosity and growth rate was investigated in two hatchery-propagated stocks of the shrimp Penaeus stylirostris, which had been genetically isolated from wild founders for 17 generations, and from each other for five more generations. presumed demographic history of these populations suggested that they were maintained at small effective population sizes. and this was confirmed by significant changes in allelic frequencies between the two stocks. Despite a small sample size (n = 60) and a small number of loci screened, a significant positive correlation was detected between microsatellite tri-locus heterozygosity and growth rate in one of the two stocks (r = 0.3, P = 0.02). In the other stock (n = 48), the positive correlation was not significant, but the combined test of single locus heterozygote advantage over the three loci was significant (P = 0.005).Use of DNA microsatellite markers, whose selectively neutral status is generally accepted, allowed us to reject the hypothesis of direct overdominance at marker loci as an explanation for an association of growth rate with heterozygosity. The average depression associated with one locus in our case is above or equivalent to the amount observed for a 10% increase of inbreeding in other species. These results suggest that heterozygosity at neutral marker loci is sufficiently well correlated with individual inbreeding coefficients to reveal a significant residual inbreeding load for growth rate in Tahitian P. stylirostris stocks. Inbreeding during hatchery propagation was, thus, insufficient to purge the ancestral load. Aquaculture (0044-8486) (Elsevier), 2000-04 , Vol. 184 , N. 3-4 , P. 203-219 Droits : 2000 Elsevier Science B.V. All rights reserved http://archimer.ifremer.fr/doc/2000/publication-753.pdf DOI:10.1016/S0044-8486(99)00331-2 http://archimer.ifremer.fr/doc/00000/753/ | Partager Voir aussi Penaeus stylirostris Heterozygosity Marker associated heterosis Inbreeding depression Microsatellites Télécharger |
Residual genetic variability in domesticated populations of the Pacific blue shrimp (Litopenaeus stylirostris) of New Caledonia, French Polynesia and Hawaii and some management recommendations Auteur(s) : Goyard, Emmanuel Arnaud-haond, Sophie Vonau, Vincent Bishoff, Vincent Mouchel, Olivier Pham, Dominique Wyban, Jim Boudry, Pierre Éditeur(s) : Elsevier Résumé : The Latin American shrimp Litopenaeus stylirostris was introduced in three different Pacific islands (Tahiti, New Caledonia via Tahiti, and Hawaii) and hatchery-propagated for 7-25 generations to develop shrimp farming based on these domesticated stocks. Three microsatellite markers have been used in an attempt to assess the genetic bases of the populations available to start a selective breeding program. The comparison of eight hatchery stocks (five New Caledonian, two Hawaiian and one Tahitian stocks) and one wild Ecuadorian population showed a much lower variability in the domesticated stocks than in the wild population, especially in New Caledonia and Tahiti (2-3.7 vs. 14-27 alleles per locus; 20-60% vs. 90% expected heterozygosity). The Tahitian and the New Caledonian stocks share the same alleles, suggesting that the loss of alleles occurred during the common past of these populations. On the contrary, New Caledonian and Hawaiian populations share only one common allele at the three loci studied. Although the low genetic variability and the resulting inbreeding of the New Caledonian stocks do not seem to affect their present performance, the results of this study demonstrate the usefulness of the introduction of new stocks in order to increase the potential responses to new controlled or uncontrolled selective pressures. The introduction in New Caledonia of the Hawaiian domesticated stocks, which would provide the local shrimp industry with 40% of the allelic diversity of the species, is advised and preferred to the one of wild animals in order to take advantage (i) of the spontaneous selection which occurred during domestication and (ii) of their favourable sanitary "specific pathogen free" status (no presence of four viruses: WSV, YHV, IHHNV, TSV) which limits the risk of introduction of pathogens. La crevette d'Amérique latine Litopaenus stylirostris a été introduite dans trois îles du Pacifique (à Tahiti, en Nouvelle-Calédonie via Tahiti, et à Hawaii), et a été ensuite reproduite en écloserie pendant 7 à 25 générations à des fins d'aquaculture. Trois marqueurs microsatellites ont été utilisés pour évaluer les bases génétiques des populations disponibles pour le démarrage d'un programme d'amélioration génétique. L'étude comparative de 8 populations domestiquées (cinq néo-calédoniennes, deux hawaiiennes et une tahitienne) et d'une population sauvage d'Equateur révèle une variabilité très réduite dans les populations d'élevage, en particulier en Nouvelle-Calédonie et à Tahiti (2 à 3.7 allèles par locus au lieu de 14 à 27 en population sauvage ; 20% à 60% d'hétérozygotie au lieu de 90%). Les souches tahitiennes et calédoniennes disposent des mêmes allèles, ce qui suggère que la perte d'allèles a eu lieu lors de l'histoire commune des ces populations. A l'inverse, les populations néo-calédoniennes et hawaiiennes n'ont en commun qu'un seul allèle sur les 3 locus étudiés. Bien que la très faible variabilité génétique du cheptel calédonien ne semble pas affecter ses performances actuelles, les résultats de cette étude démontrent l'utilité de l'introduction de variabilité afin d'augmenter la capacité de réponse à de nouvelles pressions de sélection (contrôlées ou non). L'introduction des souches hawaiiennes en Nouvelle-Calédonie qui permettrait à la filière locale de disposer de 40% de la diversité allélique de l'espèce) est préconisée de préférence à celle d'animaux sauvages afin de bénéficier (i) de la sélection spontanée qu'elles ont subi lors de leur domestication et (ii) de leur statut sanitaire « specific pathogen free, SPF » (absence de 4 virus : WSV, YHV, IHHNV, TSV) qui limite les risques de transferts de pathogènes. Aquatic Living Resources (0990-7440) (Elsevier), 2003-12 , Vol. 16 , N. 6 , P. 501-508 Droits : 2003 Published by Elsevier, Paris. http://archimer.ifremer.fr/doc/2003/publication-596.pdf DOI:10.1016/j.aquliv.2003.07.001 http://archimer.ifremer.fr/doc/00000/596/ | Partager |
Génotypage des géniteurs de Lates calcarifer de Tahiti : aide à la domestication raisonnée du Loup Tropical pour la filière tahitienne Auteur(s) : Vonau, Vincent Rouxel, Catherine Saulnier, Denis Cochennec-laureau, Nathalie Nedelec, Georges Goyard, Emmanuel Résumé : The tropical seabass Lates calcarifer was introduced in Tahiti in 1984 and then was domesticated without other introduction. Three successive generations have been obtained in captivity. Tissue samples of the 38 animaIs which represent the total Tahitian broodstock were preserved in alcohol to be genotyped with four microsatellite markers (Yue et al., 2001).
Three of four described markers were successfully transferred to the laboratory of genetics of Tahiti (marker LcaM01, LcaM02, LcaM03), as the revelation technology available locally did not allow the use of LcaM04 in routine.
The results show that the genetic diversity of the tahitian broodstock is not equivalent of the one of the species, which could be explained by the small size of the founder population, which is consistant with the history of the Tahitian population. The structuration of the 4th generation into several batches obtained from different parents appears to be the minimum management procedure to limit inbreeding. Le loup tropical Lates calcarifer a été introduit à Tahiti en 1984 puis domestiqué sans apport d'animaux extérieurs. Trois générations successives ont été obtenues en captivité. Des prélèvements de tissus sur les 38 géniteurs qui constituent l'intégralité du stock de reproducteurs tahitiens sont conservés dans l'alcool pour être génotypés à l'aide de quatre marqueurs microsatellites (Yue et al., 2001). Trois des quatre marqueurs décrits ont pu être transférés avec succès au laboratoire de génétique de Tahiti (marqueurs LcaM01, LcaM02, LcaM03), la technique de révélation disponible localement ne permettant pas d'utiliser LcaM04 en routine. Les résultats montrent qu'on ne dispose pas à Tahiti de toute la variabilité génétique de l'espèce, ce qui pourrait s'expliquer, compte tenu de l'histoire de cette population, par un faible nombre de géniteurs fondateurs. La structuration de la 4ème génération en plusieurs lots issus de parents différents apparaît comme la mesure de gestion minimale pour limiter la consanguinité au sein de cette population. Droits : 2003 Ifremer http://archimer.ifremer.fr/doc/00142/25300/23372.pdf http://archimer.ifremer.fr/doc/00142/25300/ | Partager |