Phenotypic, serological and genetic characterization of Pseudomonas anguilliseptica strains isolated from cod, Gadus morhua L., in northern Europe (original) (raw)
Abstract
The biochemical, serological and genetic characteristics of six strains of Pseudomonas anguilliseptica isolated from cod, Gadus morhua, in Scotland were compared to well characterized isolates of this same bacterial species but of different origin. Biochemical and physiological analyses showed that this group of isolates was highly homogeneous, their characteristics matching previous descriptions of the pathogen. Similar results were obtained for the six cod isolates in the serological assays, all of them belonging to the serotype O1. Marked homogeneity was observed also in the genetic study, analysed by means of RAPD, ERIC-PCR and REP-PCR procedures, showing that they were similar to isolates from gilthead seabream, Sparus aurata, black spot seabream, Pagellus bogaraveo, and turbot, Psetta maxima. Virulence assays demonstrated that the cod isolates were highly pathogenic for turbot and sole, Solea senegalensis, with LD50 between 7.6 × 104 and 5 × 107 bacterial cells per fish.
Figures (6)
Table 1 Pseudomonas anguilliseptica strains used in the study Phenotypic test
Figure 1 Application of the specific Pseudomonas anguilliseptica PCR to cod isolates. Lanes: MW, 50-2000 bp ladder molecular size marker (Sigma); 1, P. anguilliseptica CECT 399! 2, P. anguilliseptica TW-P1; 3, P. anguilliseptica AZ 208.2; 4, P. anguilliseptica Cl 3.1: 5-10, Scottish isolates TW 150/04, TW 151/04, TW 152/04, TW 153/04, TW 154/04, TW 155/04; 11, negative control (no DNA). Numbers indicate size in bp of the marker (left) and the specific fragment (right). Intensive farming of cod is just beginning, so knowledge of potential pathogens for this fish species is limited. In 2004, the first pseudomoniasis outbreaks in cod from the UK and Canada were reported (Ferguson et al. 2004). The causative agent of this disease, P. anguilliseptica, is considered a limiting factor for the culture of a number of fish species of great economic importance (Barnabe 1990; Doménech, Fernandez-Garayzabal, Lawson, Garcia, Cutuli, Blanco, Gibello, Moreno, Collins & Dominguez 1997; Romalde et al. 2003; Lopez- Romalde et al. 2003a). In this study, we character- ized a representative group of six Scottish isolates of cod, presumptively identified as P. anguilliseptica. RAPD analysis showed that these new isolates from cod had the same profile, one which is similar to that produced by Spanish gilthead seabream, black spot seabream and turbot isolates (Fig. 2a). ERIC-PCR and REP-PCR results were similar to those obtained with RAPD. Using REP-PCR, cod
Table 2 Results of biochemical characterization of cod isolates in comparison with previous descriptions of Pseudomonas anguilliseptica NC, no colour change. “Positive results only when employing Thornley semisolid medium.
Figure 2 Genetic characterization of cod isolates of Pseudomonas anguilliseptica: (a) RAPD profiles, (b) REP-PCR profiles and (c) ERIC-PCR profiles. Lanes: MW, molecular weight; 1, P. anguilliseptica CECT 399'; 2, P. anguilliseptica TW-P1; 3, P. anguilliseptica AZ 208.2; 4, P. anguilliseptica CI 3.1: 5-10, Scottish isolates TW 150/04, TW 151/04, TW 152/04, TW 153/04, TW 154/04, TW 155/04; 11, negative control (no DNA). Numbers on the left indicate size in bp of the marker.
Table 3 Results of virulence of Pseudomonas anguilliseptica in turbot and sole The results obtained by biochemical techniques, using both plate and tube tests as well as miniatur- ized API systems, showed that the isolates were
Figure 3 Moribund fish showing typical signs (arrows) of pseudomoniasis after inoculation of Pseudomonas anguilliseptica: note swollen abdomen in sole (a) and ventral haemorrhages in turbot (b).
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