Diakite, A. | Search bioRxiv

bioRxiv2026

Continental-scale survey of Treponema pallidum in African nonhuman primates: expanding the range to the Mediterranean basin

Background: Treponema pallidum, the causative agent of yaws, bejel, and syphilis, is a significant pathogen affecting both humans and nonhuman primates (NHPs). While its presence is documented in several African NHP species, the continental-scale distribution, ecological drivers, and status in North African populations remain poorly understood. We conducted a large-scale, non-invasive survey to investigate the circulation of T. pallidum across African NHP populations and evaluate geographical and ecological patterns of exposure. Results: A total of 1,696 fecal samples from 11 wild NHP species collected across five African countries were screened for anti-T. pallidum antibodies using an automated chemiluminescent Simple Western assay, and fecal and dipteran samples were analyzed by molecular methods. Widespread seropositivity was observed across all African sites and multiple primate taxa, including great apes and cercopithecids. We provide the first evidence of T. pallidum antibodies in Barbary macaques (Macaca sylvanus) from North Africa, extending the known distribution of the pathogen to the Mediterranean basin. In West Africa, chimpanzees (Pan troglodytes verus) from Senegal and Guinea exhibited particularly high seroprevalence, confirming this region as a major endemic hotspot. In Central Africa, substantial circulation was documented in Cercopithecus wolfi, Cercopithecus ascanius, and western lowland gorillas (Gorilla gorilla gorilla). Arid/savanna biomes showed significantly higher exposure than Mediterranean and tropical rainforest biomes. No T. pallidum DNA was detected in fecal or dipteran samples. Conclusion: Our findings demonstrate widespread circulation of T. pallidum among African NHPs, reveal its presence in North African primate populations for the first time. The observed high seroprevalence, coupled with significant variations across species and geographical regions, suggests the existence of multiple transmission pathways adapted to specific host behaviors and environmental constraints. These results support the existence of a broad multihost sylvatic reservoir and provide new insights into the ecology of T. pallidum, with important implications for wildlife health, One Health surveillance, and yaws eradication efforts.

microbiology↗

bioRxiv2025

Validation and multi-site deployment of a lyophilized qRT-PCR reagent for the molecular diagnosis of avian influenza and rabies in Sub-Saharan African regions

Molecular methods are widely accepted as gold standard techniques for the laboratory diagnosis of most human and animal pathogens. However, most molecular protocols rely on reagents that need to be transported and stored at a freezing temperature, a requirement that might affect their reliability in areas where the cold chain cannot be guaranteed. Over the years, several lyophilized molecular products have been marketed to circumvent this issue. We therefore evaluated the feasibility of replacing liquid reagents with freeze-dried formulations for the molecular diagnosis of avian influenza (AIV) and rabies (RABV) viruses, two priority zoonotic pathogens widely spread in Sub-Saharan Africa. Among the available kits, we selected the Qscript lyo 1-step kit (Quantabio) due to its easy-to-use features, single-reaction format, and preliminary performance assessment. Through a more in-depth evaluation, we determined its analytical and diagnostic performance and formulation stability, and obtained results comparable to those of standard liquid master mixes. Notably, for the detection of divergent lyssaviruses, the lyophilized reagents sensitivity was affected by suboptimal complementarity between the oligonucleotides and the target sequences. Finally, a multi-site evaluation in four veterinary diagnostic laboratories located in Sub-Saharan Africa demonstrated the successful deployment of AIV and RABV assays utilizing freeze-dried reagents, which can interchangeably replace liquid master mixes. Altogether our results indicate that the Qscript lyo 1-step kit (Quantabio) represents a valid alternative to wet reagents for the molecular diagnosis of avian influenza and rabies, and has the potential for broader applications to other relevant infectious diseases upon proper validation. Author summaryMolecular diagnostic protocols rely on reagents that need to be transported and stored at freezing temperatures. Meeting this requirement can be challenging in areas where the maintenance of the cold chain is not guaranteed, such as in sub-Saharan Africa. Our study aimed to assess the feasibility of using lyophilized reagents as a replacement for liquid reagents in the molecular diagnosis of two widespread zoonotic pathogens in Sub-Saharan Africa, namely avian influenza and rabies. To accomplish this, we selected a commercially available lyophilized reagent based on its format and performance characteristics. We conducted a laboratory validation to assess the use of the lyophilized reagent throughout the entire diagnostic process. We also conducted a reproducibility test involving African laboratories as potential end-users. Our findings confirm that the lyophilized reagent can replace traditional liquid reagents to diagnose rabies and avian influenza and suggest its possible use for a wider range of infectious diseases after undergoing appropriate validation.

microbiology↗