Spotted Fever Group Rickettsiae Research Articles

Rickettsia sp. and Anaplasma spp. in Haemaphysalis longicornis from Shandong province of China, with evidence of a novel species “Candidatus Anaplasma Shandongensis”

Haemaphysalis longicornis is one of the most dominant and widespread tick species in China. This species mainly infests wild animals and occasionally attacks humans, and has been associated with the transmission of a variety of zoonotic pathogens including spotted fever group Rickettsia (SFGR), severe fever with thrombocytopenia syndrome virus (SFTSV), Anaplasma phagocytophilum, Babesia spp. and Theileria spp.. Although there are increasing reports of various pathogens associated with H. longicornis, some neglected pathogens in certain areas still need to be studied. In this study, a total of 171 H. longicornis ticks were collected from goats in three locations of Shandong Province, Eastern China (Zibo, Linyi, and Qingdao cities), and subsequently screened for the presence of Rickettsia, Anaplasma, and Ehrlichia bacteria. A total of four bacterial species were identified and characterized. “Candidatus Rickettsia jingxinensis” was detected in one tick specimen from Zibo city. Of 98 ticks from Linyi city, 63.27% (62/98) were tested positive for Anaplasma capra and 5.10% (5/98) were positive for Anaplasma bovis. Interestingly, a novel Anaplasma species was detected and characterized in one tick specimen from Zibo and one other from Linyi, respectively. Genetic and phylogenetic analysis based on the 16S, gltA, groEL, and msp4 genes indicated that it was divergent from all known Anaplasma species but mostly related to A. phagocytophilum and “Cadidatus Anaplasma boleense”. Based on where it was first detected, we named it “Candidatus Anaplasma shandongensis”.

Serosurvey of spotted fever group Rickettsia in equids from western Pará, Amazon, Brazil.

From a previous large epidemiological survey, we randomly selected 474 serum samples (463 horses and 11 mules) distributed among four municipalities of Pará state, Amazon region, Brazil, and from three types: farm animal, urban carthorse, and sport horse. Samples were tested by indirect fluorescence antibody test (IFAT ≥ 64) for antibodies reactive to spotted fever group (SFG) rickettsiae using Rickettsia rickettsii as crude antigens. From the 474 equids tested, 149 (31.4%) had ticks attached during sampling, belonging mostly to the species Dermacentor nitens. The overall seroprevalence for SFG rickettsiae was 31.4% (95% confidence interval: 27.3-35.9%) with 149 seropositive animals out of 474 screened. Notably, 77 equids (16.2%) had high endpoint titers ranging from 512 to 16,384, indicating that they had been exposed to SFG rickettsiae not long before sampling. Animal type affected rickettsial seroprevalence, with significantly higher values among farm horses when compared with urban and sport animals. Presence of dogs and tick infestation were negatively associated with equid seropositivity to R. rickettsii. This is the first report of SFG rickettsiae-reactive antibodies in equids from Pará state, Brazilian Amazon.

Evaluating the Clinical and Immune Responses to Spotted Fever Rickettsioses in the Guinea Pig-Tick-Rickettsia System.

The guinea pig was the original animal model developed for investigating spotted fever rickettsiosis (SFR). This model system has persisted on account of the guinea pig's conduciveness to tick transmission of SFR agents and ability to recapitulate SFR in humans through clinical signs that include fever, unthriftiness, and in some cases the development of an eschar. The guinea pig is the smallest animal model for SFR that allows the collection of multiple blood and skin samples antemortem for longitudinal studies. This unit provides the basic protocols necessary to establish, maintain, and utilize a guinea pig-tick-Rickettsia model for monitoring the course of infection and immune response to an infection by spotted fever group Rickettsia (SFGR) that can be studied at biosafety level 2 (BSL-2) and arthropod containment level 2 (ACL-2); adaptations must be made for BSL-3 agents. The protocols cover methods for tick feeding and colony development, laboratory infection of ticks, tick transmission of Rickettsia to guinea pigs, and monitoring of the course of infection through clinical signs, rickettsial burden, and immune response. It should be feasible to adapt these methods to study other tick-borne pathogens. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Tick transmission of SFGR to guinea pigs Support Protocol 1: Laboratory infection of ticks by injection Alternate Protocol 1: Needle inoculation of SFGR to guinea pigs Basic Protocol 2: Monitoring the course of guinea pig rickettsial infection: clinical signs Basic Protocol 3: Monitoring the course of guinea pig rickettsial infection: collection of biological specimens Support Protocol 2: Guinea pig anesthesia Basic Protocol 4: Monitoring rickettsial burden in guinea pigs by multiplex qPCR Basic Protocol 5: Monitoring guinea pig immune response to infection: blood leukocytes by flow cytometry Basic Protocol 6: Monitoring immune response to guinea pig rickettsial infection: leukocyte infiltration of skin at the tick bite site by flow cytometry Basic Protocol 7: Monitoring the immune response to guinea pig rickettsial infection: antibody titer by ELISA Support Protocol 4: Coating ELISA Plates Alternate Protocol 2: Monitoring immune response to guinea pig rickettsial infection: antibody titer by immunofluorescence assay.

Procedure for spotted fever group Rickettsia isolation from limited clinical blood specimens.

BackgroundCurrent isolation techniques for spotted fever group Rickettsia from clinical samples are laborious and are limited to tissue, blood and blood derivatives with volumes ideally greater than 1 mL. We validated the use of simplified methodologies for spotted fever group Rickettsia culture isolation that overcome sample volume limitations and provide utility in clinical diagnostics and research studies.Methodology/Principal findingsA modified cell culture method is evaluated for the isolation of Rickettsia ssp. from human diagnostic samples. Culture sampling method, culture platform, and growth phase analysis were evaluated to determine best practices for optimal culture isolation conditions. Rickettsial isolates (R. conorii, R. rickettsii, and R. parkeri) were grown in Vero E6 cells over a course of 5 to 7 days at low inoculum treatments (~40 bacterial copies) to standardize the sampling strategy at a copy number reflective of the bacteremia in acute diagnostic samples. This methodology was verified using small volumes (50 μL) of 25 unprocessed clinical whole blood, plasma, and serum samples from acute samples of patients suspected of having Rocky Mountain Spotted Fever, of which 10 were previously confirmed positive via the PanR8 qPCR assay, 13 had no detectable Rickettsia DNA by the PanR8 qPCR assay, and 2 were not previously tested; these samples resulted in the cultivation of 7 new R. rickettsii isolates.Conclusions/SignificanceWe observed that rickettsial isolate growth in culture is reproducibly identified by real-time PCR testing of culture media within 72 hours after inoculation. Additionally, specimen sedimentation prior to isolation to remove red blood cells was found to decrease the amount of total organism available in the inoculum. A small volume culture method was established focusing on comparative qPCR detection rather than bacterial visualization, taking significantly shorter time to detect, and requiring less manipulation compared to traditional clinical isolate culture methods.

High diversity of Rickettsia spp., Anaplasma spp., and Ehrlichia spp. in ticks from Yunnan Province, Southwest China.

Rickettsia, Anaplasma, and Ehrlichia belonging to the order Rickettsiales are causative agents of tick-borne diseases in humans. During 2021, 434 ticks including Rhipicephalus microplus and R. haemaphysaloides were collected from three sampling sites in Yunnan Province, Southwest China, and analyzed for the presence of these bacteria. Nine bacterial species were identified, including two Rickettsia spp., three Anaplasma spp., and four Ehrlichia spp., some of which are potential human pathogens. Genetic and phylogenetic analysis on 16S rRNA, gltA, groEL, ompA, ompB, and sca4 genes indicated the presence of a novel spotted fever group Rickettsia (SFGR) named “Candidatus Rickettsia shennongii” in six of the 38 R. haemaphysaloides ticks from two locations, Dehong Autonomous Prefecture and Honghe City. Another SFGR species, Candidatus Rickettsia jingxinensis was detected in ticks from all three sites, with an overall positive rate of 62.67%. Three other human pathogenic species, Anaplasma ovis (1.38%, 6/434), Ehrlichia canis (16.36%, 71/434), and E. chaffeensis (0.23%, 1/434) were detected in these ticks and characterized. Moreover, Ehrlichia sp. (4.84%, 21/434), E. minasensis (7.37%, 32/434), A. marginale (6.91%, 30/434), and Cadidatus Anaplasma boleense (1.15%, 5/434) were detected in R. microplus ticks, for which pathogenicity to humans remains to be determined. The results reveal the remarkable diversity of Rickettsiales bacteria in ticks from Yunnan Province, Southwest China. The high infection rate of some human pathogenic bacteria in ticks may indicate potential infection risk in humans, and it highlights the need for surveillance in local populations.

Diversity and geographic distribution of rickettsial agents identified in brown dog ticks from across the United States

Rhipicephalus sanguineus sensu lato, or brown dog ticks, transmit a variety of pathogens of veterinary and public health importance globally. Pathogens vectored by brown dog ticks and identified in the United States include Babesia vogeli, Ehrlichia canis, and several spotted fever group Rickettsia spp. (SFGR). Due to the challenge of collecting canine blood samples nationwide to screen for exposure to these pathogens, we took an indirect approach and tested brown dog ticks for molecular evidence of infection. Brown dog ticks (616 adults and 65 nymphs) collected from dogs and cats across the nation were tested by separate PCR assays detecting Babesia spp., E. canis, and SFGR. While no Babesia sp. was found, we identified rickettsial agents in 3.5% (24/681; 95% CI 2.4–5.2%) of the ticks. Pathogens and related organisms detected in ticks included E. canis (n = 1), Rickettsia amblyommatis (n = 3), Rickettsia massiliae (n = 11), Rickettsia monacensis (n = 3), Rickettsia montanensis (n = 5), and an undefined Rickettsia species (n = 1). These data demonstrate a wider geographic distribution of R. massiliae than previously known, and to the authors’ knowledge, reports R. monacensis in brown dog ticks for the first time. Due to the close association that brown dog ticks have with domestic dogs and humans, more research is needed to understand the full array of organisms, some of which are zoonotic, potentially transmitted by this widespread tick complex.

Coinfection of Two Rickettsia Species in a Single Tick Species Provides New Insight into Rickettsia-Rickettsia and Rickettsia-Vector Interactions.

ABSTRACTRickettsiae are obligate intracellular bacteria that can cause life-threatening illnesses. There is an ongoing debate as to whether established infections by one Rickettsia species preclude the maintenance of the second species in ticks. Here, we identified two Rickettsia species in inoculum from Haemaphysalis montgomeryi ticks and subsequently obtained pure isolates of each species by plaque selection. The two isolates were classified as a transitional group and spotted fever group rickettsiae and named Rickettsia hoogstraalii str CS and Rickettsia rhipicephalii str EH, respectively. The coinfection of these two Rickettsia species was detected in 25.6% of individual field-collected H. montgomeryi. In cell culture infection models, R. hoogstraalii str CS overwhelmed R. rhipicephalii str EH with more obvious cytopathic effects, faster plaque formation, and increased cellular growth when cocultured, and R. hoogstraalii str CS seemed to polymerize actin tails differently from R. rhipicephalii str EH in vitro. This work provides a model to investigate the mechanisms of both Rickettsia-Rickettsia and Rickettsia-vector interactions.IMPORTANCE The rickettsiae are a group of obligate intracellular Gram-negative bacteria that include human pathogens causing an array of clinical symptoms and even death. There is an important question in the field, that is whether one infection can block the superinfection of other rickettsiae. This work demonstrated the coinfection of two Rickettsia species in individual ticks and further highlighted that testing the rickettsial competitive exclusion hypothesis will undoubtedly be a promising area as methods for bioengineering and pathogen biocontrol become amenable for rickettsiae.

Molecular Detection of Tick-Borne Bacteria from Amblyomma (Acari: Ixodidae) Ticks Collected from Reptiles in South Africa.

Reptiles are hosts for various tick species and tick-associated organisms, many of which are zoonotic. However, little is known about the presence and diversity of tick-borne bacteria infecting reptiles and their ticks in South Africa. Amblyomma ticks (n = 253) collected from reptiles were screened for the presence of Coxiella, Anaplasma, Rickettsia, and Borrelia species by amplification, sequencing and phylogenetic analysis of the 16S rRNA, 23S rRNA, gltA, OmpA, and Flagellin genes, respectively. This study recorded the presence of reptile associated Borrelia species and Coxiella-like endosymbiont in South Africa for the first time. Furthermore, a spotted fever group Rickettsia species was observed in 7 Amblyomma marmoreum and 14 Amblyomma sylvaticum from tortoises of genera Kinixys and Chersina. Francisella-like endosymbiont was observed from 2 Amblyomma latum collected from the Mozambique spitting cobra, Naja mossambica. Coxiella burnetii and Anaplasma spp., were not detected from the current samples. Although the direct evidence that reptiles can act as reservoir hosts remains to be determined, observations from this study provide indications that reptilian ticks may play a role in the transmission of pathogenic bacteria to homothermic animals. Furthermore, the absence of Anaplasma spp., and C. burnetii does not mean that these pathogens should be completely neglected.

Visual closed dumbbell-mediated isothermal amplification (CDA) for on-site detection of Rickettsia raoultii.

Spotted fever group (SFG) rickettsioses are important zoonoses, threatening human health seriously and gradually attracting more attention in the world. SFG rickettsiae are classified as neglected pathogens. If these pathogens are detected at all, they are usually recognized very late in the infection through indirect detection of specific antibodies. Previous studies have shown that Rickettsia raoultii (R. raoultii), a member of the SFG rickettsiae, occurs with increasing incidence in remote countries. Therefore, a rapid detection method for R. raoultii is in urgently need. In this study, a R. raoultii diagnosis method by closed dumbbell-mediated isothermal amplification (R-CDA) assay targeting a conserved sequence of the outer membrane protein A (OmpA) gene with high sensitivity and specificity was developed. This assay offered a rapid and simple method for on-site detection of R. raoultii. Firstly, four pairs of R-CDA primers were designed and the optimum primer set was selected to amplify target gene specifically and effectively. Then, a pair of outer primer was designed to accelerate the reaction based on the inner primers to establish the RO-CDA reaction. In addition, the results of real-time amplification curves, melting curves and end-point colorimetric judgements showed that the established visual RO-CDA reaction could accurately detect R. raoultii without cross-reaction with other closely related pathogens. Furthermore, the detection limit of visual RO-CDA assay was 10 copies/μL, which was feasible for on-site detection with merits of easy-operation, rapidity, high sensitivity, and specificity. In conclusion, the developed RO-CDA detection method could be helpful for pathogen screening and epidemic prevention at the point of care.

Human case of Anaplasma phagocytophilum infection in Eastern Taiwan

Human granulocytic anaplasmosis (HGA) is a tick-borne infection caused by the bacterium Anaplasma phagocytophilum. In this study, we report an indigenous case of clinically diagnosed HGA. The patient was a 41-year-old man who experienced a tick bite and later developed fever, chills, myalgia, malaise, thrombocytopenia, leukocytosis with a left shift, elevated hepatic transaminase levels, and splenomegaly upon admission to the hospital. Immunofluorescence assays detected seroconversion against A. phagocytophilum, whereas tests for spotted fever group rickettsiae, murine typhus, scrub typhus, Q fever, and ehrlichiosis were negative. ELISA and Western blot analysis using recombinant MSP2 protein confirmed the exposure to A. phagocytophilum. Oral doxycycline and intravenous ceftriaxone were prescribed, and the patient made a full recovery. Our findings indicate the presence of HGA on the main island of Taiwan. Precautions against tick bites should be taken when engaging in outdoor activities, and HGA should be considered by physicians in the differential diagnosis.

Unraveling the epidemiological relationship between ticks and rickettsial infection in Africa

Tick-borne rickettsioses are emerging and re-emerging diseases of public health concern caused by over 30 species ofRickettsia. Ticks are obligate hematophagous arthropods with over 700 species of Ixodid ticks known worldwide. The escalating geographical dispersal of tick vectors and concomitant increase in the incidences of tick-borne diseases have fueled interest in the ecology of tick-borne pathogens. This review focuses on aspects of theRickettsiapathogen, including biology, taxonomy, phylogeny, genetic diversity, epidemiology of the disease, and the role of vertebrate host in the perpetuation of rickettsioses in Africa. Our review also highlights some of the species ofRickettsiathat are responsible for disease, the role of tick vectors (both hard and soft ticks) and the species ofRickettsiaassociated with diverse tick species across the continent. Additionally, this article emphasizes the evolutionary perspective of rickettsiae perpetuation and the possible role of amplifying vertebrate host and other small mammals, domestic animals and wildlife in the epidemiology ofRickettsiaspecies. We also specifically, discussed the role of avian population in the epidemiology of SFG rickettsiae. Furthermore, we highlighted tick-borne rickettsioses among travelers due to African tick-bite fever (ATBF) and the challenges to surveillance of rickettsial infection, and research on rickettsiology in Africa. Our review canvasses the need for more rickettsiologists of African origin based within the continent to further research towards understanding the biology, characterization, and species distribution, including the competent tick vectors involved in their transmission of rickettsiae across the continent in collaboration with established researchers in western countries. We further highlighted the need for proper funding to encourage research despite competing demands for resources across the various sectors. We finalize by discussing the similarities between rickettsial diseases around the world and which steps need to be taken to help foster our understanding on the eco-epidemiology of rickettsioses by bridging the gap between the growing epidemiological data and the molecular characterization ofRickettsiaspecies.

Molecular detection of spotted fever group rickettsiae in hedgehogs (Erinaceusamurensis) and hedgehog-attached ticks in Xuyi County, Southeast China

Tick-borne diseases like Rickettsia, Anaplasma and Ehrlichia are widespread infectious zoonoses that threaten the health of both humans and animals worldwide. Ticks and their hosts, such as hedgehogs, can play a crucial role in transmitting tick-borne diseases and the cycle of Rickettsia. To investigate the presence and identity of Rickettsia in hedgehogs and hedgehog-attached ticks in Xuyi County, Southeast China, 114 ticks were collected from 45 hedgehogs captured totally. Via morphological and molecular methods, all these ticks were identified as two species: Haemaphysalis flava (110/114, 96.5%) and Haemaphysalis longicornis (4/114, 3.5%). Rickettsia spp. were genotypically characterized by PCR targeting rrs, gltA, ompA, ompB, and sca4 gene fragments. The prevalence of spotted fever group rickettsiae (SFGR) infection found in hedgehogs and ticks was 17.8% (8/45) and 78.1% (89/114), respectively. Phylogenetic analyses demonstrated that those Rickettsia spp. belong to two species: Rickettsia heilongjiangensis (R. heilongjiangensis XY-1) and a potential new species, Candidatus Rickettsia xuyiensis XY-2. The present study gave the first evidence of R. heilongjiangensis and Candidatus R. xuyiensis in ticks and hedgehogs of Southeast China. Our findings suggest that hedgehogs might be involved in the natural transmission cycle of Rickettsia species.

Incidence of tick-borne spotted fever group Rickettsia species in rodents in two regions in Kazakhstan

Records on the distribution of Rickettsia spp. in their natural hosts in Central Asia are incomplete. Rodents and small mammals are potential natural reservoirs for Rickettsiae in their natural lifecycle. Studies about the maintenance of Rickettsia in wild animals are available for Western nations, but—to our knowledge—no studies and data are available in the Republic of Kazakhstan so far. The first case description of Rickettsioses in Kazakhstan was made in the 1950ies in the Almaty region and now Kyzylorda, East Kazakhstan, Pavlodar and North Kazakhstan are endemic areas. The existence of murine and endemic typhus was proven in arthropod vectors in the regions Kyzylorda and Almaty. Here we show for the first time investigations on tick-borne Rickettsia species detected by a pan-rickettsial citrate synthase gene (gltA) real-time PCR in ear lobes of small mammals (n = 624) in Kazakhstan. From all analysed small mammals 2.72% were positive for Rickettsia raoultii, R. slovaca or R. conorii. Sequencing of the rickettsial gene OmpAIV and the 23S–5S interspacer region revealed a similar heritage of identified Rickettsia species that was observed in ticks in previous studies from the region. In summary, this study proves that rodents in Kazakhstan serve as a natural reservoir of Rickettsia spp.

Total RNA sequencing of Phlebotomus chinensis sandflies in China revealed viral, bacterial, and eukaryotic microbes potentially pathogenic to humans

ABSTRACT Phlebotomus chinensis sandfly is a neglected insect vector in China that is well-known for carrying Leishmania. Recent studies have expanded its pathogen repertoire with two novel arthropod-borne phleboviruses capable of infecting humans and animals. Despite these discoveries, our knowledge of the general pathogen diversity and overall microbiome composition of this vector species is still very limited. Here we carried out a meta-transcriptomics analysis that revealed the actively replicating/transcribing RNA viruses, DNA viruses, bacteria, and eukaryotic microbes, namely, the “total microbiome”, of several sandfly populations in China. Strikingly, “microbiome” made up 1.8% of total non-ribosomal RNA and comprised more than 87 species, among which 70 were novel, including divergent members of the genera Flavivirus and of the family Trypanosomatidae. Importantly, among these microbes we were able to reveal four distinguished types of human and/or mammalian pathogens, including two phleboviruses (hedi and wuxiang viruses), one novel Spotted fever group rickettsia, as well as a member of Leishmania donovani complex, among which hedi virus and Leishmania each had > 50% pool prevalence rate and relatively high abundance levels. Our study also showed the ubiquitous presence of an endosymbiont, namely Wolbachia, although no anti-viral or anti-pathogen effects were detected based on our data. In summary, our results uncovered the much un-explored diversity of microbes harboured by sandflies in China and demonstrated that high pathogen diversity and abundance are currently present in multiple populations, implying disease potential for exposed local human population or domestic animals.

Ixodid diversity and detection of spotted fever group Rickettsia spp. in ticks collected on birds in the Brazilian Atlantic Forest

The Brazilian Atlantic Forest helds one of the most diverse and unique avifauna in the world. Many vertebrate species are reservoirs of tick-borne pathogens, and birds are an important group among them due to their mobility which facilitates the dispersion of ticks and the infectious agents they carry. This study brings data on the tick diversity parasitizing birds and the molecular detection of Rickettsia spp. in these arthropods. Birds (n = 773) were captured, identified, and banded at Mata do Paraíso Research, Training, and Environmental Education Center located in Viçosa, Minas Gerais, Brazil. Birds were checked for the presence of ticks, which were individually collected, identified, and molecularly processed through Polymerase Chain Reaction (PCR) for the detection of Rickettsia spp. A total of 130 individuals were infested by ticks, and 479 tick specimens were collected, showing a seasonal distribution of the life stages throughout the year. Ticks were identified as Amblyomma longirostre (59/479); Amblyomma calcaratum (20/479); Amblyomma varium (3/479); Amblyomma sculptum (2/479) and Amblyomma spp. larvae (395/479). Seasonal distribution of the life stages of ticks was observed along the year and significant negative correlations were found between temperature and collected ticks and temperature and infested birds. From the evaluated samples of ticks, 25.44% (n = 43/169) scored positive for Rickettsia spp., and sequence analysis indicated high nucleotide identity with Rickettsia rhipicephali, R. massiliae, R. africae and R. honei marmionii. The potential for dispersal of ticks by birds added to the aggressiveness of species of the genus Amblyomma and the zoonotic potential of some species of Rickettsia are quite worrying when we consider that the study area is widely attended by students, researchers, people from the city and neighboring municipalities.

Increasing Incidence of Spotted Fever Group Rickettsioses in the United States, 2010-2018.

Spotted fever group Rickettsia species are intracellular bacteria transmitted by tick or mite vectors and that cause human diseases referred to as spotted fever group rickettsioses, or spotted fevers. In the United States, the most recognized and commonly reported spotted fevers are Rocky Mountain spotted fever (RMSF) (Rickettsia rickettsii), Rickettsia parkeri rickettsiosis, Pacific Coast tick fever (Rickettsia species 364D), and rickettsialpox (Rickettsia akari). In this study, we summarize and evaluate surveillance data on spotted fever cases reported to the Centers for Disease Control and Prevention (CDC) through the National Notifiable Diseases Surveillance System from 2010 to 2018. During this period, there were 36,632 reported cases of spotted fevers with 95.83% (N = 35,104) reported as meeting the case definition as probable and 4.17% (N = 1528) reported as meeting the case definition as confirmed. The average national incidence of total cases, both probable and confirmed, was 12.77 cases per million persons per year. The highest statewide incidence was in Arkansas, with 256.84 per million per year, whereas the lowest incidence occurred in California, with 0.32 per million per year (note that spotted fevers were not notifiable in Hawaii and Alaska). Cases of spotted fevers were reported more frequently among males by gender, White by race, and non-Hispanic by ethnicity. The incidence of spotted fevers increased significantly from 2010 to 2018, but it is uncertain how many of the reported cases were RMSF and how many developed from more moderate spotted fevers. Improvement of the ability to differentiate between spotted fever group Rickettsia species is needed.

First detection and molecular identification of a pathogenic spotted fever group Rickettsia, R. massiliae, from Rhipicephalus haemaphysaloides ticks infesting dogs in southern Taiwan

Tick-borne Rickettsia pathogens become an emerging zoonotic infection worldwide. The prevalence and genetic identity of Rickettsia infection was determined firstly in Rhipicephalus haemaphysaloides ticks collected from dogs in southern Taiwan. A total of 141 Rh. haemaphysaloides ticks were examined for Rickettsia infection by nested-PCR assay targeting the citrate synthase (gltA) and outer membrane protein B (ompB) genes of Rickettsia. The Rickettsia infection was detected with a general infection rate of 2.84%, and was detected in male and female ticks with an infection rate of 3.13% and 2.60%, respectively. Genetic relationships were analyzed by comparing the gltA and ompB sequences obtained from 4 Taiwan strains and 15 other strains representing 13 genospecies of Rickettsia. Phylogenetic analyses reveal that all Taiwan strains were genetically affiliated with the R. massiliae (spotted fever group) and can be distinguished from other genospecies of Rickettsia. These results demonstrate the epidemiological significance of a human pathogenic Rickettsia species (R. massiliae) detected in Rh. haemaphysaloides ticks. Further study focused on the vector competence of this tick species may help to illustrate the potential threat for human infection in southern Taiwan.

Rickettsia-Host-Tick Interactions: Knowledge Advances and Gaps.

ABSTRACTTicks are hematophagous ectoparasites capable of transmitting multiple human pathogens. Environmental changes have supported the expansion of ticks into new geographical areas that have become the epicenters of tick-borne diseases (TBDs). The spotted fever group (SFG) of Rickettsia frequently infects ticks and causes tick-transmitted rickettsioses in areas of endemicity where ixodid ticks support host transmission during blood feeding. Ticks also serve as a reservoir for SFG Rickettsia. Among the members of SFG Rickettsia, R. rickettsii causes Rocky Mountain spotted fever (RMSF), the most lethal TBD in the United States. Cases of RMSF have been reported for over a century in association with several species of ticks in the United States. However, the isolation of R. rickettsii from ticks has decreased, and recent serological and epidemiological studies suggest that novel species of SFG Rickettsia are responsible for the increased number of cases of RMSF-like rickettsioses in the United States. Recent analyses of rickettsial genomes and advances in genetic and molecular studies of Rickettsia provided insights into the biology of Rickettsia with the identification of conserved and unique putative virulence genes involved in the rickettsial life cycle. Thus, understanding Rickettsia-host-tick interactions mediating successful disease transmission and pathogenesis for SFG rickettsiae remains an active area of research. This review summarizes recent advances in understanding how SFG Rickettsia species coopt and manipulate ticks and mammalian hosts to cause rickettsioses, with a particular emphasis on newly described or emerging SFG Rickettsia species.

Comprehensive screening of tick-borne microorganisms indicates that a great variety of pathogens are circulating between hard ticks (Ixodoidea: Ixodidae) and domestic ruminants in natural foci of Anatolia

Grazing domestic ruminants serve as important reservoirs and/or amplificatory hosts in the ecology of tick-borne pathogens (TBPs) and tick vectors in the natural foci; however, many enzootic life cycles including ruminants and ticks are still unknown. This study investigated a wide range of TBPs circulating among ticks and grazing ruminants in the natural foci of Anatolia, Turkey. Tick specimens (n = 1815) were collected from cattle, sheep, and goats in three ecologically distinct areas (wooded, transitional, and semi-arid zones) of Anatolia and identified by species: Dermacentor marginatus, Dermacentor reticulatus, Hyalomma anatolicum, Hyalomma excavatum, Hyalomma marginatum, Hyalomma scupense, Haemaphysalis inermis, Haemaphysalis parva, Haemaphysalis punctata, Haemaphysalis sulcata, Ixodes ricinus, Rhipicephalus bursa, and Rhipicephalus turanicus. PCR-sequencing analyses revealed TBPs of great diversity, with 32 different agents identified in the ticks: six Babesia spp. (Babesia occultans, Babesia crassa, Babesia microti, Babesia rossi, Babesia sp. tavsan1, and Babesia sp. Ucbas); four Theileria spp., including one putative novel species (Theileria annulata, Theileria orientalis, Theileria ovis, and Theileria sp.); one Hepatozoon sp.; four Anaplasma spp., including one novel genotype (Anaplasma phagocytophilum, Anaplasma marginale, Anaplasma ovis, and Anaplasma sp.); six unnamed Ehrlichia spp. genotypes; Neoehrlichia mikurensis; nine spotted fever group rickettsiae, including one putative novel species (Rickettsia aeschlimannii, Rickettsia slovaca, Rickettsia hoogstraalii, Rickettsia monacensis with strain IRS3, Rickettsia mongolitimonae, Rickettsia raoultii, Candidatus Rickettsia goldwasserii, Candidatus Rickettsia barbariae, and Rickettsia sp.); and Borrelia valaisiana. Detailed phylogenetic analyses showed that some of the detected pathogens represent more than one haplotype, potentially relating to the tick species or the host. Additionally, the presence of Neoehrlichia mikurensis, an emerging pathogen for humans, was reported for the first time in Turkey, expanding its geographical distribution. Consequently, this study describes some previously unknown tick-borne protozoan and bacterial species/genotypes and provides informative epidemiological data on TBPs, which are related to animal and human health, serving the one health concept.

Spotted fever group Rickettsia, Anaplasma and Coxiella-like endosymbiont in Haemaphysalis ticks from mammals in Thailand

Ticks are ectoparasites of vertebrates and vectors of various pathogenic microorganisms. In this study, the presence of bacteria and protozoa was evaluated by PCR and DNA sequencing in 233 mammal ticks collected from 8 provinces in Thailand. Sequence and phylogenetic analyses of partial rickettsial ompA, ompB, sca4 and partial Coxiella 16S rRNA, GroEL, rpoB genes clearly revealed, for the first time, a co-infection of SFG Rickettsia belonging to R. massiliae subgroup and Coxiella-like endosymbiont (CLE), Cox-hein, in a male of Haemaphysalis heinrichi tick infesting Burmese ferret-badger in Loei province. Moreover, a male of H. hystricis tick infesting the same host was infected with another CLE, Cox-hys. Based on the 16S rRNA gene sequence, Anaplasma sp., closely related to Anaplasma bovis was also detected in a male of H. heinrichi infesting the same Burmese ferret-badger. In addition, the third CLE, Cox-asia, found in H. asiatica collected from Asian palm civet in Chiang Rai province, was different from both Cox-hein and Cox-hys. This study provided important data and broadened our knowledge on tick-borne pathogens and endosymbionts in Thailand and Southeast Asia.