Prasetyo, Didot Budi, Fiorenzano, Jodi M., Nop, Daliya, Noch, Nin, Huot, Boren, Mom, Sokly, Prum, Sitha, Chhe, Visal, Dul, Sokha, Heang, Vireak, Prom, Satharath, Jiang, Ju, Richards, Allen L., Farris, Christina M., and Hertz, Jeffrey C.
Arthropod-borne rickettsioses comprise a wide variety of subtypes that are endemic in Cambodia, but there remains very little data on the geographic distribution of the pathogens or their vectors. Surveys were conducted in Koh Kong and Preah Sihanouk Provinces between September 2017 and June 2018 to collect ectoparasites from peridomestic animals and the environment using dragging and flagging methods. Collected ectoparasites were sorted and identified morphologically, then pooled by species, host, and location for molecular detection using Rickettsia genus- and species-specific qPCR and/or multilocus sequence typing (MLST) assays. A total of 14,254 ectoparasites were collected including seven new locality records. Rickettsia species were detected in 35.5% (174/505) of the pools screened representing 3,149 randomly selected ectoparasites from the total collected. Rickettsia asembonensis was detected in 89.6% (147/164) of Rickettsia-positive flea pools and 3.6% (6/164) of the flea pools were positive for both R. asembonensis and Rickettsia felis. Candidatus Rickettsia senegalensis from Ctenocephalides orientis fleas and Rickettsia sp. close to Rickettsia japonica and Rickettsia heilongjiangensis from Haemaphysalis ticks were identified by MLST. This appears to be the first report of these new ectoparasite records and rickettsial species in southern Cambodia, suggesting a potential health risk to military and civilians in this region. Author summary: Rickettsiae are Gram-negative bacteria that cause acute undifferentiated febrile illness in humans. These pathogens are associated with arthropods and primarily transmitted by ectoparasites such as ticks, fleas, and mites. In Cambodia, rickettsial diseases have been documented in local patients, travelers and military personnel. However, the lack of entomological studies and awareness of these diseases hindered our comprehensive understanding of these pathogens and their arthropod vectors. Our manuscript provides a description of ectoparasite diversity and sheds light on their potential role in rickettsial disease transmission. Through molecular techniques, we uncover the prevalence and diversity of Rickettsia species in ectoparasites collected from various animal hosts in Koh Kong and Preah Sihanouk province. Rickettsia asembonensis, Rickettsia felis, and Candidatus Rickettsia senegalensis were all identified from Ctenocephalides orientis fleas, while Rickettsia species closely related to Rickettsia japonica, Rickettsia heilongjiangensis, and Rickettsia raoultii were found from Haemaphysalis ticks. Additionally, Rickettsia were also detected in Heterodoxus spiniger lice and Laelapidae mites, although we were unable to characterize the species. Insights into this pathogen-vector-host interface will better inform healthcare professionals, leading to more accurate diagnoses and increased awareness for better management of these diseases. [ABSTRACT FROM AUTHOR]