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. 2020 Dec 1;70(6):487-491.
doi: 10.30802/AALAS-CM-99-990085. Epub 2020 Oct 29.

Differentiation Among Rodentibacter Species Based on 16S-23S rRNA Internal Transcribed Spacer Analysis

Laurentiu Benga  1 , Peter M Benten  2 , Eva Engelhardt  2 , Karl Köhrer  3 , Barbara Hueber  4 , Werner Nicklas  5 , Henrik Christensen  6 , Martin Sager  2
Affiliations

Differentiation Among Rodentibacter Species Based on 16S-23S rRNA Internal Transcribed Spacer Analysis

Laurentiu Benga et al. Comp Med. .

Abstract

The internal transcribed spacer (ITS) regions of Rodentibacter pneumotropicus, R. heylii, R. rarus, R. ratti, and R. heidelbergensis and of a Rodentibacter- related β-hemolytic Pasteurellaceae taxon isolated from laboratory rodents were studied for their feasibility to discriminate among these species. The 6 species analyzed showed species-specific ITS patterns that were shared by the type strains and clinical isolates and that allowed their identification. Nevertheless, differentiating between the ITS band patterns of R. pneumotropicus and R. ratti is visually challenging. In all species tested, sequence analysis of the ITS fragments revealed a larger ITSile+ala, which contained the genes for tRNAIle(GAU) and tRNA Ala(UGC), and a smaller ITSglu with the tRNAGlu(UUC) gene. The ITS sequences varied among the 6 species evaluated, displaying identity levels ranging from 62% to 86% for ITSile+ala and 68% to 90% for ITSglu. Overall, ITS amplification proved to be a reliable method to differentiate among these important Pasteurellaceae species of laboratory rodents. Moreover, the ITS sequence variations recorded here might facilitate the design of probes for specific identification of these species. The ability to diagnose these organisms to the species level could increase our understanding of their clinical significance.

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Figures

Figure 1.
Figure 1.
The ITS PCR-amplification patterns of (A) type strains and (B) clinical isolates of Rodentibacter spp. (A) Lanes 1 and 8, 100-bp size standards; lane 2, R. heylii (ATCC12555T); lane 3, R. pneumotropicus (P421T); lane 4, R. rarus (CCUG17206T); lane 5, R. ratti (F75T); lane 6, R. heidelbergensis (Ac69T); and lane 7, β-hemolytic Pasteurellaceae (1725/19). (B) Lanes 1 and 17, 100-bp ladder; lanes 2 through 4, R. heylii (594/17, 220/17, 1338/18); lanes 5 through 7, R. pneumotropicus (646/19, 443/17b, 620/19); lanes 8 through 10, R. ratti (257/18, 260/18, 638/17); lanes 11 through 13, R. heidelbergensis (H201108024, Ac79, H199605711); and lanes 14 through 16, β-hemolytic Pasteurellaceae (1725/19, 1625/19, 268/20).
See this image and copyright information in PMC

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