Rapid and sensitive PCR detection of Vibrio penaeicida, the putative etiological agent of syndrome 93 in New Caledonia

Type Article
Date 2000-03
Language English
Author(s) Saulnier Denis, Avarre Jean-Christophe, Le Moullac GillesORCID, Ansquer Dominique, Levy Peva, Vonau Vincent
Affiliation(s) IFREMER, Ctr Oceanol Pacifique, Lab Aquaculture Trop, Taravao 98719, Tahiti, Fr Polynesia.
Source Diseases of aquatic organisms (0177-5103) (Inter-Research), 2000-03 , Vol. 49 , N. 2 , P. 109-115
DOI 10.3354/dao040109
WOS© Times Cited 49
Keyword(s) small subunit rDNA gene, Penaeus (Litopenaeus) stylirostris, vibriosis, shrimp disease, PCR diagnosis, vibrio penaeicida, Syndrome 93
Abstract 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.
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Saulnier Denis, Avarre Jean-Christophe, Le Moullac Gilles, Ansquer Dominique, Levy Peva, Vonau Vincent (2000). Rapid and sensitive PCR detection of Vibrio penaeicida, the putative etiological agent of syndrome 93 in New Caledonia. Diseases of aquatic organisms, 49(2), 109-115. Publisher's official version : https://doi.org/10.3354/dao040109 , Open Access version : https://archimer.ifremer.fr/doc/00000/4436/