Molecular detection and characterization of novel haemotropic Mycoplasma in free-living mole rats from South Africa

Abstract

The importance of haemotropic Mycoplasma (haemoplasma) infections to animal and human health is increasingly recognised. Although wild rodents are known to host these bacteria, haemoplasma prevalence and diversity in small mammals is under-documented, globally. This is due to the reliance on molecular approaches to detect these unculturable, obligate bacteria and to a paucity of assays targeting informative gene regions. We attempted to address these challenges by evaluating the performance of three 16S rRNA PCR assays for detecting Mycoplasma in four African mole-rat species of the family Bathyergidae. This was achieved by screening DNA samples prepared from lung and liver samples of 260 bathyergids, sampled from natural and urban landscapes in the Western Cape Province with one published and two novel conventional PCR assays. Sequence-confirmed Mycoplasma presence guided calculations of the relative sensitivity and specificity of the assays and revealed that 26.5% of the rodents were haemoplasma-positive. Bathyergus suillus sampled near an informal human settlement had a significantly higher infection rate (42%) than the three bathyergid species sampled from natural settings, for which PCR-positivity ranged from 0% to 36%. The 16S rRNA gene phylogeny identified the presence of six Mycoplasma strains in bathyergids that form a novel monophyletic lineage belonging to the haemofelis group, with 16S rRNA and Rnase P gene phylogenies indicating that the bathyergid-associated haemoplasmas were novel and closely related to Mycoplasma coccoides. Assay sensitivity ranged from 60.3% to 76.8% and specificity from 94.8% to 100% and both were highest for the novel assay targeting a ~ 300 bp region of the 16S rRNA gene. Results confirm the presence of novel haemoplasma strains in bathyergid species from South Africa and emphasise the need for expanded studies on haemoplama prevalence, diversity, and transmission routes in other small mammal species from this biodiverse region.

Keywords: 16S rRNA; Mycoplasma coccoides; PCR; Phylogeny; RNase P; Sequencing.