Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Culture-Independent PCR Detection and Differentiation of Mycobacteria spp. in Antemortem Respiratory Samples from African Elephants (Loxodonta africana) and Rhinoceros (Ceratotherium simum, Diceros bicornis) in South Africa

Version 1 : Received: 27 May 2022 / Approved: 30 May 2022 / Online: 30 May 2022 (16:33:30 CEST)

A peer-reviewed article of this Preprint also exists.

Goosen, W.J.; Clarke, C.; Kleynhans, L.; Kerr, T.J.; Buss, P.; Miller, M.A. Culture-Independent PCR Detection and Differentiation of Mycobacteria spp. in Antemortem Respiratory Samples from African Elephants (Loxodonta africana) and Rhinoceros (Ceratotherium simum, Diceros bicornis) in South Africa. Pathogens 2022, 11, 709. Goosen, W.J.; Clarke, C.; Kleynhans, L.; Kerr, T.J.; Buss, P.; Miller, M.A. Culture-Independent PCR Detection and Differentiation of Mycobacteria spp. in Antemortem Respiratory Samples from African Elephants (Loxodonta africana) and Rhinoceros (Ceratotherium simum, Diceros bicornis) in South Africa. Pathogens 2022, 11, 709.

Abstract

Since certain Mycobacterium tuberculosis complex (MTBC) members, like M. bovis, are endemic in specific South African wildlife reserves and zoos, cases of clinically important nontuberculous mycobacteria (NTM) in wildlife may be neglected. Additionally, due to the inability of tests to differentiate between host responses to MTBC and NTM, the diagnosis of MTBC may be confounded by the presence of NTMs. This may hinder control efforts. These constraints highlight the need for enhanced rapid detection and differentiation methods for MTBC and NTM, especially in high MTBC burden areas. We evaluated the use of the GeneXpert MTB/RIF Ultra, the Hain CMdirect V1.0, and novel amplicon sequencing PCRs targeting the mycobacterial rpoB and ku gene targets, directly on antemortem respiratory samples from known MTBC-infected and NTM culture-positive African elephants (n=26 animals) and rhinoceros (n=23 animals). Our findings suggest that the Ultra is the most sensitive diagnostic test for MTBC DNA detection directly in raw antemortem respiratory specimens and that the rpoB PCR is ideal for Mycobacterium genus DNA detection and species identification through amplicon sequencing.

Keywords

African elephants; broncholoalveolar lavage; Hain CMdirect V1.0; ku PCR; Mycobacterium tuberculosis complex; non-tuberculous mycobacteria; rhinoceros; rpoB PCR; trunk wash; Xpert MTB/RIF Ultra

Subject

Medicine and Pharmacology, Veterinary Medicine

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