| Summary: | The soft ticks, <i>Ornithodoros sonrai</i>, are known as vectors of the tick-borne relapsing fever caused by <i>Borrelia</i> spp. and have also been reported to carry other micro-organisms. The objective of this study was to collect and to identify <i>O. sonrai</i> ticks and to investigate the micro-organisms associated with them. In 2019, an investigation of burrows within human dwellings was conducted in 17 villages in the Niakhar area and in 15 villages in the Sine-Saloum area in the Fatick region of Senegal. Ticks collected from the burrows were identified morphologically and by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Micro-organism screening was performed by bacteria-specific qPCR and some identifications were made by standard PCR and gene sequencing. <i>O. sonrai</i> ticks were found in 100% (17/17) of the villages surveyed in the Niakhar area and in 66% (10/15) of the villages in the Sine-Saloum area. A total of 1275 soft tick specimens were collected from small mammal burrows. The ticks collected were morphologically identified as <i>O. sonrai</i>. About 20% (259/1275) of the specimens were also submitted to MALDI-TOF MS for identification. Among the resulting MS profiles, 87% (139/159) and 95% (95/100) were considered good quality specimens, preserved in alcohol and silica gel, respectively. All spectra of good quality were tested against our MALDI-TOF MS arthropod spectra database and identified as <i>O. sonrai</i> species, corroborating the morphological classification. The carriage of four micro-organisms was detected in the ticks with a high prevalence of <i>Bartonella</i> spp., Anaplasmataceae, and <i>Borrelia</i> spp. of 35, 28, and 26%, respectively, and low carriage of <i>Coxiella burnetii</i> (2%). This study highlights the level of tick infestation in domestic burrows, the inventory of pathogens associated with the <i>O. sonrai</i> tick, and the concern about the potential risk of tick involvement in the transmission of these pathogens in Senegal.
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