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The 16S-23S RNA gene intergenic spacers of isolates of  Streptococcus equi  ( n =5),  S. zooepidemicus  ( n =5),  S. equisimilis  ( n =3) and  S. dysgalactiae  ( n =2) were sequenced and compared. There were distinct regions within the spacer, arranged in the order 1–9 for all  S. equi  and one  S. zooepidemicus  isolate and 1,2 and 4–9 for the remaining isolates. Region 4 was identical to the tRNA ala  gene found in the 16S-23S intergenic spacers of other streptococci. Regions 1, 5, 6 and 7 had distinct variations, each conserved in different isolates. However, amongst the intergenic spacers there were different combinations of variant regions, suggesting a role for DNA recombination in their evolution. The intergenic spacer of all isolates of  S. equi  and one  S. zooepidemicus  isolate were almost identical. Primers derived from the variant sequences of regions 1 and 5 to 6 were used to group all  S. zooepidemicus  ( n =17) and  S. equi  ( n =5) into 1 of 8 types by polymerase chain reaction; three  S. zooepidemicus  isolates typed the same as  S. equi .  S. equi  and  S. zooepidemicus  were clearly distinguishable from  S. equisimilis  and  S. dysgalactiae  which had shorter regions 5 and 6 and no region 7. Most homology for the group C sequences was found in previously published sequences for the 16S-23S intergenic spacers of  S. anginosis ,  S. constellatus ,  S. intermedius ,  S. salivarius  and  S. agalactiae . A 75-90 nucleotide length shared with  S. anginosus  and  S. intermedius  in opposite orientations in the two main variants of region 6 supported the role for DNA recombination in the evolution of the spacer. The 16S-23S intergenic spacers indicate that  S. zooepidemicus  was the archetypal species for  S. equi  and that both are genetically more distant from  S. equisimilis  and  S. dysgalactiae . The intergenic spacer can be used to identify specifically the group C. streptococci and as an epidemiological marker for  S. zooepidemicus .

epidemiology and infection

Characterization of the Lancefield group C streptococcus 16S-23S RNA gene intergenic spacer and its potential for identification and sub-specific typing