Use of an exclusion assay to detect novel rickettsiae in field collected Amblyomma americanum.

In the south-central United States, several tick-borne diseases (TbDs) occur at or near their highest levels of incidence of anywhere in the U.S. The diversity of Rickettsia species found in Amblyomma americanum continues to be under-characterized in this region and throughout the U.S. and Canada where this tick species is expanding. One reason for this lack of knowledge about Rickettsia diversity is the high prevalence of the endosymbiont Rickettsia amblyommatis that obscures detection of other bacteria in this genus. Focusing on unknown rickettsial agents, we used a recently described R. amblyommatis exclusion assay to screen 1909 A. americanum collected in Oklahoma City, Oklahoma, which resulted in eight ticks that had unique rickettsial sequences. Through the process of characterizing primary and secondary rickettsiae, we identified ticks primarily infected with Rickettsia rhipicephali and a Rickettsia species (2019-CO-FNY) previously linked with a canine rickettsiosis case in Tulsa, Oklahoma. We also identified a Rickettsia agent that was 97% identical with an endosymbiont of Amblyomma tonelliae and which aligned with archaic rickettsial species. Through this study, we further demonstrate the usefulness of this exclusion assay for rapid screening in large cohort A. americanum studies to identify a small number of ticks that contain poorly described and previously undocumented rickettsiae.

Seasonality of Ticks and Prevalence of Rickettsiae Species in Dermacentor variabilis and Amblyomma maculatum Across Oklahoma Pastures.

Tick-borne diseases are an increasing concern for people and companion animals in the United States, but there is a need for continued vigilance regarding livestock in pasture systems. The south-central United States has some of the highest incidences of tick-borne diseases, and there is a need to re-examine the ecology of tick vectors in relation to pasture systems and livestock. The objective of this study was to establish a baseline of seasonal activity for tick species in diverse regional Oklahoma pastures and screen for important pathogens in Dermacentor variabilis (Say) and Amblyomma maculatum Koch group that may impact livestock and human health. Between 2015 and 2017, transects in five pastures across Oklahoma were visited each month. DNA extracted from adult D. variabilis and A. maculatum group was tested for the presence of bacterial pathogens. We found that tick communities in pastures across Oklahoma differ by season, abundance, and bacterial presence and prevalence. The peak abundance of Amblyomma americanum (L.) adults and nymphs occurred a month earlier over the 2 yr of the study compared with historical studies in the same regions. Additionally, we observed notable differences in peak activity between A. americanum adults and nymphs collected in pastures in central Oklahoma (April) versus pastures in northern part of the state (May). We detected Rickettsia parkeri, R. bellii, and Anaplasma sp. DNA in D. variabilis from pastures across the state. These results potentially have important ramifications for human and livestock risk of encountering infected ticks in pastures across the southern Great Plains.