All Group II strains are non-proteolytic and include type E strains and some type B and type F strains. Nucleotide sequencing of various toxin genes has demonstrated the presence of amino acid variation within genes encoding a single toxin serotype and these variants are identified as toxin subtypes [9, 10]. Among type E strains, a Dasatinib in vivo total of 8 such

subtypes (E1-E8) have been identified [11]. These subtypes differ at the amino acid level by up to 6%. The genes encoding BoNT/A-G are found in toxin gene clusters that also encode several nontoxic proteins and regulatory proteins. The gene encoding BoNT/E is found within a toxin gene cluster that includes ntnh (nontoxic nonhemagglutinin), p47, and orfX1-3[12, 13]. Hill et al. [13] demonstrated that the bont/E toxin gene cluster inserted into the rarA selleck inhibitor operon. The transposon-associated gene, rarA, likely plays a role in this insertion event in which the gene is split into small and large fragments that flank the toxin gene cluster [13]. Remarkably, an intact rarA gene is also located within the toxin gene cluster and the nucleotide sequences of the intact and split genes were shown to differ by phylogenetic analysis. Moreover, the split rarA gene fragments can be pasted together to form a gene with a nucleotide sequence with similarity CHIR-99021 molecular weight to the gene found in the Group II C. botulinum type B strain 17B. In another study, the intact and split rarA genes

were detected across a panel of 41 type E strains [11]. In this study, we characterized a previously unreported C. botulinum type E strain isolated Molecular motor in 1995 from soil in Chubut, Argentina. This represents the first report of a type E strain (CDC66177) originating from the Southern hemisphere. We further show evidence that this strain produces a unique type E toxin subtype and that the genetic background of this strain is highly divergent compared

to other type E strains. Results and discussion Phylogenetic analysis of bont/E in C. botulinum strains The nucleotide sequence of the entire bont/E gene was determined for each of the 16 C. botulinum type E strains examined in this study. Previous studies have identified several bont/E subtypes [9–12]. Nucleotide sequences of bont/E determined in this study were compared along with representatives of other reported bont/E subtypes (Figure 1). It is important to note that in some cases strain names used in previous reports may not refer to identical strains examined in this study with a similar name. For instance, the CDC reference strain labeled “Alaska” harbored a gene encoding a subtype E2 toxin and is unlikely to be related to the genome-sequenced strain Alaska E43 (Genbank accession number: NC_010723) which encodes a subtype E3 toxin. Another strain labeled “Minnesota” was distinguished from a strain with the same name reported by Macdonald et al. [11].