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BackgroundCoxiella bacteria were identified from various tick species across the world. Q fever is a zoonotic disease caused by the bacteria Coxiella burnetii that most commonly infects a variety of mammals. Non-mammalian hosts, such as birds, have also been reported to be infected with the pathogenic form of “Candidatus Coxiella avium”. This research increases the list of tick species that have been found with Coxiella-like bacteria in Thailand.MethodsA total of 69 ticks were collected from 27 domestic fowl (Gallus gallus domesticus), 2 jungle fowl (Gallus gallus) and 3 Siamese firebacks (Lophura diardi) at 10 locations (provinces) in Thailand. Ticks were identified and PCR was used to amplify Coxiella bacteria 16S rRNA, groEL and rpoB genes from the extracted tick DNA. MEGA6 was used to construct phylogenetic trees via a Maximum Likelihood method.ResultsThe phylogenetic analysis based on the 16S rRNA gene showed that the Coxiella sequences detected in this study grouped in the same clade with Coxiella sequences from the same tick genus (or species) reported previously. In contrast, rpoB gene of the Coxiella bacteria detected in this study did not cluster together with the same tick genus reported previously. Instead, they clustered by geographical distribution (Thai cluster and Malaysian cluster). In addition, phylogenetic analysis of the groEL gene (the chaperonin family) showed that all Coxiella bacteria found in this study were grouped in the same clade (three sister groups).ConclusionsTo our knowledge, we found for the first time rpoB genes of Coxiella-like bacteria in Haemaphysalis wellingtoni ticks forming two distinct clades by phylogenetic analysis. This may be indicative of a horizontal gene transfer event.

Tick-borne viruses and bacteria that can cause diseases of animals and humans have high impact and are of concern as significant threats to human health worldwide. In this research, we screened microorganisms related to those pathogens in ticks from dogs, a cat, and a cow. The techniques used were PCR, DNA sequencing and phylogenetic analysis to detect and classify the microorganisms [Flavivirus, severe fever with thrombocytopenia syndrome virus (SFTSV), Phlebovirus, Coronavirus, Canine Parvovirus, eubacteria, Coxiella and Rickettsia]. A novel virus named Phlebovirus-like-AYUT and Stenotrophomonas maltophilia bacteria were found in one individual tick (Rhipicephalus sanguineus s.l.) from a dog. All tick samples were negative for Rickettsia, while 9/21 (42.9 %) were positive for Coxiella bacteria. The novel virus “Phlebovirus-like-AYUT” (the name derives from Phra Nakhon Si Ayutthaya Province in Thailand) was resolved by phylogenetic analysis of the partial L segment by maximum likelihood (ML) method using MEGA X. The phylogenetic tree also indicated that the virus was related to Phlebovirus in brown dog ticks reported in Trinidad and Tobago. In contrast, Phlebovirus-like-AYUT was in a distinct clade from Lihan tick Phlebovirus-Thailand (LTPV), which was previously found in cow ticks, Rhipicephalus microplus, in Nan Province, Thailand. This study reports the Stenotrophomonas maltophilia bacterium with a novel Phlebovirus-like-AYUT in a brown dog tick. The roles of this bacterium in a virus-positive tick or in viral transmission from animal host requires further investigation.

Ticks can transmit a wide variety of pathogens, including bacteria. Here, we report the detection of tick-associated bacteria in Chaiyaphum Province, northeastern Thailand. There have been few reports of tick-borne bacterial pathogens in the study areas, which are evergreen forests dominated by plateaus at elevations of approximately 1,000 m. In total, 94 ticks were collected from vegetation. They were screened for the presence of Coxiella, Francisella, Rickettsia, and Borrelia bacteria using PCR assays. In this study, we found ticks from two genera, Haemaphysalis and Amblyomma, that were positive for Coxiella-like bacteria (CLB) and Rickettsia. Francisella and Borrelia spp. were not detected in these two tick genera. The results revealed the evolutionary relationships of CLB in Amblyomma testudinarium, Haemaphysalis lagrangei, and Haemaphysalis obesa ticks using the 16S rRNA and rpoB markers, which clustered together with known isolates of ticks from the same genera. In contrast, the groEL marker showed different results. On the basis of the groEL phylogenetic analysis and BLAST results, three groups of CLB were found: (1) CLB from A. testudinarium grouped as a sister clade to CLB from Ixodes ricinus; (2) CLB from Haemaphysalis lagrangei was distantly related to CLB from Haemaphysalis wellingtoni; and (3) CLB from A. testudinarium grouped as sister clade to CLB from Amblyomma from French Guiana and Brazil. For Rickettsia studies, phylogenetic trees of the gltA, ompB, and sca4 genes revealed two groups of Spotted Fever Group (SFG) Rickettsiae: (1) SFG Rickettsiae that formed a sister clade with Rickettsia tamurae AT-1 (belong to the Rickettsia helvetica subgroup) in A. testudinarium and (2) SFG Rickettsiae that formed a distantly related group to Rickettsia rhipicephali 3-7-female6-CWPP (belong to the Rickettsia massiliae subgroup) in A. testudinarium. This study expanded our knowledge of the diversity of tick-borne Coxiella and Rickettsia bacteria. The pathogenic roles of these bacteria also need to be investigated further.