Characterization of yellow-pigmented and motile enterococci isolated from intestines of the garden snail Helix aspersa P. S ˇ vec 1 , L.A. Devriese 2 , I. Sedla ´c ˇek 1 , M. Baele 2 , M. Vancanneyt 3 , F. Haesebrouck 2 , J. Swings 3 and J. Dos ˇkar ˇ 4 1 Czech Collection of Micro-organisms, Masaryk University, Brno, Czech Republic, 2 Laboratory of Bacteriology and 3 BCCM/LMG Bacteria Collection, Ghent University, Belgium, and 4 Department of Genetics and Molecular Biology, Masaryk University, Brno, Czech Republic 2001/144: received 16 June 2001, revised 29 November 2001 and accepted 19 December 2001 P. S ˇ VEC, L.A. DEVRIESE, I. SEDLA ´ C ˇ EK, M. BAELE, M. VANCANNEYT, F. HAESEBROUCK, J. SWINGS AND J. DOS ˇ KAR ˇ . 2002. Aims: Enterococci associated with garden snails (Helix aspersa) were studied in order to obtain reliable species identification and characterization. Methods and Results: Twelve yellow-pigmented and motile enterococci, isolated from the intestines of garden snails, were phenotypically close to Enterococcus casseliflavus, but they showed certain unusual biochemical characteristics. tRNA intergenic length polymorphism analysis (tDNA-PCR) divided all strains studied into two groups, in full agreement with biochemical test results. 16S rDNA sequencing, DNA base composition analysis and DNA- DNA hybridization results showed unambiguously that the enterococci studied belonged to the species Ent. casseliflavus. The representative strains of described ecovars were deposited in the Czech Collection of Microorganisms (CCM) as Ent. casseliflavus CCM 4868, 4869, 4870 and 4871. Conclusions: Enterococcus casseliflavus associated with garden snails can be subdivided into groups. Significance and Impact of the Study: Enterococcus casseliflavus differs from other enterococcal species in that it is typically associated with plants, soil, water and invertebrate animals. The different groups that can be found in these widely occurring bacteria are possibly source-specific ecovars, as exemplified by the Ent. casseliflavus inhabiting the intestines of snails. INTRODUCTION Enterococci are Gram-positive, catalase-negative cocci com- monly isolated from various sources. Nowadays, they play an important role in human clinical microbiology, mainly because of their frequent occurrence as nosocomial infec- tious agents as well as their increasing acquired antibiotic resistance (Murray 1990; Facklam et al. 1999). Most taxo- nomic studies on these bacteria have been based on strains from human clinical material and infections. In contrast, few investigations focus on environmental strains. Enterococci can be isolated from water, soil, plants, and various animals including insects, reptiles, birds and mammals. These environment-colonizing strains are often phenotypically very heterogeneous and different from the strains isolated from human clinical materials (Devriese et al. 1993; Niemi et al. 1993; Devriese and Pot 1995; Ulrich and Mu ¨ller 1998). In this study, 12 atypical yellow-pigmented and motile enterococci isolated from the intestines of garden snails, Helix aspersa, were analysed. Biochemical test results divided these strains into two groups, one of which was composed of two subgroups. All strains included were phenotypically close to Ent. casseliflavus (Collins et al. 1984), but they showed certain characteristics unusual for this species. Because of the unclear taxonomic position of the strains studied, tRNA intergenic length polymorphism Correspondence to: P. S ˇ vec, Czech Collection of Microorganisms, Masaryk University, Tvrde´ho 14, 602 00 Brno, Czech Republic (e-mail: mpavel@ sci.muni.cz). ª 2002 The Society for Applied Microbiology Journal of Applied Microbiology 2002, 92, 951–957