Non-invasive specimen collections for Mycobacterium tuberculosis detection in free-ranging long-tailed macaques (Macaca fascicularis)

Suthirote Meesawat; Nalina Aiempichitkijkarn; Saradee Warit; Mutchamon Kaewparuehaschai; Suchinda Malaivijitnond

doi:10.1371/journal.pone.0289961

Non-invasive specimen collections for Mycobacterium tuberculosis detection in free-ranging long-tailed macaques (Macaca fascicularis)

PLoS One. 2023 Aug 24;18(8):e0289961. doi: 10.1371/journal.pone.0289961. eCollection 2023.

Authors

Suthirote Meesawat^{1

2}, Nalina Aiempichitkijkarn³, Saradee Warit⁴, Mutchamon Kaewparuehaschai⁵, Suchinda Malaivijitnond^{2

6}

Affiliations

¹ Faculty of Science, Biological Sciences Program, Chulalongkorn University, Bangkok, Thailand.
² Faculty of Science, Department of Biology, Chulalongkorn University, Bangkok, Thailand.
³ Animal Behavior Graduate Group, University of California, Davis, California, United States of America.
⁴ Industrial Tuberculosis Team (ITBT), IMBG, BIOTEC, NSTDA, Thailand Science Park, Pathumthani, Thailand.
⁵ Department of National Parks, Wildlife Rescue Center No.2 (Krabokkoo), Wildlife and Plant Conservation, Chachoengsao, Thailand.
⁶ National Primate Research Center of Thailand-Chulalongkorn University, Saraburi, Thailand.

Abstract

Surveillance of infectious diseases in free-ranging or wild animals has been widely conducted in many habitat-range countries after the COVID-19 episode. Thailand is located in the center of the distribution range of long-tailed macaques (Macaca fascicularis; Mf) where the animals have both frequent human contact and a high prevalence of human tuberculosis. For the large-scale detection of Mycobacterium tuberculosis complex (MTBC) using IS6110-nested PCR in free-ranging Mf, non-invasive sampling was developed using oral (via rope bait) and fecal (direct swabs of fresh feces) specimen collection. Firstly, the MTBC-IS6110-nested PCR was validated in non-invasively collected specimens, in terms of its specificity and sensitivity, and then compared with those of the invasively collected oral and rectal swabs in 24 captive MTBC-suspected Mf. After validation, these methods were applied to survey for the prevalence of shed MTBC (MTBCS) in four previously reported MTBC-infected populations. A total of 173 baited rope specimens and 204 freshly defecated excretions were collected. The limit of detection of the IS6110-nested PCR technique was 10 fg/μL and the 181-bp PCR amplicon showed 100% sequence similarity with the MTB H37Rv genome sequence. Comparing the MTBCS detection between the invasive and non-invasive collected specimens in captive suspected Mf revealed a significant correlation between the two types of oral specimens (oral swabs and baited ropes; n = 24, r2 = 1, p-value < 0.001), but fresh fecal swabs showed higher MTBCS frequencies than the rectal swabs. Moreover, the proportion of MTBCS-positive free-ranging Mf were significantly higher in the fresh fecal swabs (8.82%; 95% CI; 4.9-12.7%) than in the baited ropes (5.20%; 95% CI; 1.9-8.5%). This result indicates that oral sampling via baited ropes and fecal sampling via defecated excretion swabs can serve as ancillary specimens for MTBCS detection in free-ranging non-human primates.

Copyright: © 2023 Meesawat et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Wild
COVID-19*
Macaca fascicularis
Mycobacterium tuberculosis* / genetics
Specimen Handling

Grants and funding

This research was supported by the Research Fund Senior Scholar (grant number RTA6280010 to SM), the TSRI Fund (to SM), the NSRF via the Program Management Unit for Human Resources & Institutional Development, Research and Innovation (grant number B05F640122 to SM), the 90th Anniversary of Chulalongkorn University Ratchadaphiseksomphot Endowment Fund (to SM), and Chulalongkorn University - NSTDA Doctoral Scholarship (to SM). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.