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. 2000 Apr;66(4):1544-52.
doi: 10.1128/AEM.66.4.1544-1552.2000.

Diversity of Salmonella strains isolated from the aquatic environment as determined by serotyping and amplification of the ribosomal DNA spacer regions

J Baudart  1 , K Lemarchand, A Brisabois, P Lebaron
Affiliations

Diversity of Salmonella strains isolated from the aquatic environment as determined by serotyping and amplification of the ribosomal DNA spacer regions

J Baudart et al. Appl Environ Microbiol. 2000 Apr.

Abstract

Salmonella species are pathogenic bacteria often detected in sewage, freshwater, marine coastal water, and groundwater. Salmonella spp. can survive for long periods in natural waters, and the persistence of specific and epidemic strains is of great concern in public health. However, the diversity of species found in the natural environment remains unknown. The aim of this study was to investigate the diversity of Salmonella strains isolated from different natural aquatic systems within a Mediterranean coastal watershed (river, wastewater, and marine coastal areas). A total of 574 strains isolated from these natural environments were identified by both conventional serotyping and the ribosomal spacer-heteroduplex polymorphism (RS-HP) method (M. A. Jensen and N. Straus, PCR Methods Appl. 3:186-194, 1993). More than 40 different serotypes were found, and some serotypes probably mobilized from widespread animal-rearing activities were detected only during storm events. These serotypes may be good indicators of specific contamination sources. Furthermore, the RS-HP method based on the PCR amplification of the intergenic spacer region between the 16S and 23S rRNA genes can produce amplicon profiles allowing the discrimination of species at both serotype and intraserotype levels. This method represents a powerful tool that could be used for rapid typing of Salmonella isolates.

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Figures

FIG. 1
FIG. 1
Study areas. (a) The Tech River watershed and location of sampling sites. (b) Location of the wastewater treatment station of Banyuls-sur-Mer.
FIG. 2
FIG. 2
Representative electrophoretic patterns of PCR-amplified bacterial genomic DNA isolated from the indicated Salmonella spp. serotypes, with RS-HP types indicated in brackets. (A) Lane 1, serotype Newport (New4); lane 2, serotype Newport (New3); lane 3, serotype Newport (New2); lane 4, serotype Newport (New1); lane 5, serotype Kottbus (Kott2); lane 6, serotype Kottbus (Kott1). (B) Lane 1, serotype Typhimurium (Typ1); lane 2, serotype Typhimurium (Typ2); lane 3, serotype Oranienburg; lane 4, serotype Bardo; lane 5, serotype Indiana; lane 6, serotype Muenster. (C) Lane 1, serotype Paratyphi B; lane 2, serotype Hadar; lane 3, serotype Senftenberg (Senf2); lane 4, serotype Senftenberg (Senf1); lane 5, serotype Panama (Pana2); lane 6, serotype Panama (Pana1). (D) Lane 1, serotype Derby; lane 2, serotype Gold Coast; lane 3, serotype Wien; lane 4, serotype Veneziana; lane 5, serotype Coeln; lane 6, serotype Mbandaka. (E) Lane 1, serotype Grumpensis; lane 2, serotype Virchow (Vir3); lane 3, serotype Virchow (Vir2); lane 4, serotype Virchow (Vir1); lane 5, serotype Bareilly; lane 6, serotype Braenderup. (F) Lane 1, serotype Saint-Paul; lane 2, serotype Brandenburg (Bran1); lane 3, serotype Brandenburg (Bran2); lane 4, serotype Enteritidis; lane 5, serotype Infantis; lane 6, serotype Richmond. (G) Lane 1, serotype Bredeney (Bre1); lane 2, serotype Bredeney (Bre2); lane 3, serotype Bredeney (Bre3); lane 4, serotype Anatum; lane 5, serotype Agona; lane 6, serotype Montevideo. (H) Lane 1, serotype Give; lane 2, serotype Bovis morbificans; lane 3, serotype Haifa; lane 4, serotype Mikawasima; lane 5, serotype Rissen; lane 6, serotype Kedougou. (I) Lane 1, serotype London; lane 2, serotype Manhattan; lane 3, serotype Cerro; lane 4, serotype Kapemba; lane 5, serotype III b.38: z10: z53 (“Salmonella enterica subsp. diarizonae”). PCR products were run on a 4% acrylamide-bisacrylamide gel (29:1) with a D-code electrophoresis system. Lanes without numbers, molecular-weight DNA markers (100-bp ladder).
FIG. 2
FIG. 2
Representative electrophoretic patterns of PCR-amplified bacterial genomic DNA isolated from the indicated Salmonella spp. serotypes, with RS-HP types indicated in brackets. (A) Lane 1, serotype Newport (New4); lane 2, serotype Newport (New3); lane 3, serotype Newport (New2); lane 4, serotype Newport (New1); lane 5, serotype Kottbus (Kott2); lane 6, serotype Kottbus (Kott1). (B) Lane 1, serotype Typhimurium (Typ1); lane 2, serotype Typhimurium (Typ2); lane 3, serotype Oranienburg; lane 4, serotype Bardo; lane 5, serotype Indiana; lane 6, serotype Muenster. (C) Lane 1, serotype Paratyphi B; lane 2, serotype Hadar; lane 3, serotype Senftenberg (Senf2); lane 4, serotype Senftenberg (Senf1); lane 5, serotype Panama (Pana2); lane 6, serotype Panama (Pana1). (D) Lane 1, serotype Derby; lane 2, serotype Gold Coast; lane 3, serotype Wien; lane 4, serotype Veneziana; lane 5, serotype Coeln; lane 6, serotype Mbandaka. (E) Lane 1, serotype Grumpensis; lane 2, serotype Virchow (Vir3); lane 3, serotype Virchow (Vir2); lane 4, serotype Virchow (Vir1); lane 5, serotype Bareilly; lane 6, serotype Braenderup. (F) Lane 1, serotype Saint-Paul; lane 2, serotype Brandenburg (Bran1); lane 3, serotype Brandenburg (Bran2); lane 4, serotype Enteritidis; lane 5, serotype Infantis; lane 6, serotype Richmond. (G) Lane 1, serotype Bredeney (Bre1); lane 2, serotype Bredeney (Bre2); lane 3, serotype Bredeney (Bre3); lane 4, serotype Anatum; lane 5, serotype Agona; lane 6, serotype Montevideo. (H) Lane 1, serotype Give; lane 2, serotype Bovis morbificans; lane 3, serotype Haifa; lane 4, serotype Mikawasima; lane 5, serotype Rissen; lane 6, serotype Kedougou. (I) Lane 1, serotype London; lane 2, serotype Manhattan; lane 3, serotype Cerro; lane 4, serotype Kapemba; lane 5, serotype III b.38: z10: z53 (“Salmonella enterica subsp. diarizonae”). PCR products were run on a 4% acrylamide-bisacrylamide gel (29:1) with a D-code electrophoresis system. Lanes without numbers, molecular-weight DNA markers (100-bp ladder).
FIG. 3
FIG. 3
UPGMA dendrogram generated from the similarity matrix determined from the different RS-HP banding patterns corresponding to 574 Salmonella spp. strains.
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