Analysis of 16S–23S rRNA internal transcribed spacer regions in Pasteurellaceae isolated from laboratory rodents

Benga, Laurentiu; Benten, W. Peter M.; Engelhardt, Eva; Christensen, Henrik; Sager, Martin

Analysis of 16S–23S rRNA internal transcribed spacer regions in Pasteurellaceae isolated from laboratory rodents

2012

Benga, Laurentiu | Benten, W. Peter M. | Engelhardt, Eva | Christensen, Henrik | Sager, Martin

The internal transcribed spacer (ITS) regions of members of Pasteurellaceae isolated from rodents, including the [Pasteurella] pneumotropica biotypes Jawetz and Heyl, [Actinobacillus] muris, “Hemophilus influenzaemurium” and Bisgaard taxon 17 were studied and their feasibility to discriminate these species was analyzed. The reference strains of all species analyzed showed unique species‐specific ITS patterns which were further present in 49 clinical isolates of [P.] pneumotropica biotypes Jawetz and Heyl and [A.] muris allowing their identification by comparison to the reference strains pattern. Sequence analysis of the amplified fragments revealed in all species, with exception of “H. influenzaemurium”, a larger ITSⁱˡᵉ⁺ᵃˡᵃ which contained the genes for tRNAᴵˡᵉ⁽ᴳᴬᵁ⁾ and tRNAᴬˡᵃ⁽ᵁᴳC⁾ and a smaller ITSᵍˡᵘ with the tRNAᴳˡᵘ⁽ᵁᵁC⁾ gene. “H. influenzaemurium” revealed two each of the larger and respectively the smaller ITS fragments. Both the length and the sequence of each ITS type were highly conserved within the [P.] pneumotropica biotypes Jawetz and Heyl and [A.] muris strains tested. On the contrary, ITS sequences revealed significant interspecies variations with identity levels ranging from 61.2 to 89.5% for ITSⁱˡᵉ⁺ᵃˡᵃ and 56.5 to 86.8% for ITSᵍˡᵘ. Sequences regions with significant interspecies variation but highly conserved within the species were identified and might be used to design probes for the identification of rodent Pasteurellaceae to the species level.

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