Spotted Fever Group Rickettsia Species Research Articles

Molecular evidence and phylogenetic delineation of spotted fever group Rickettsia species in Amblyomma ticks from cattle in Gauteng and Limpopo Provinces, South Africa

Objective: To determine the prevalence of tick-borne pathogens with a particular focus on Rickettsia spp. in ticks collected from cattle in Gauteng and Limpopo Provinces, South Africa. Methods: A total of 200 ticks were collected from cattle within the Madala livestock, Pretoria, Gauteng Province and in Mankweng Township, Polokwane, Limpopo Province in 2019. The ticks were morphologically identified and processed individually for a total genomic DNA extraction. Specific primers targetting ompA, ompB, and the 17KDa genes were used for a molecular screening and delineation of Rickettsia from the extracted genetic materials using polymerase chain reaction (PCR) technique. PCR amplicons of positive samples were sequenced bidirectionally using the Sanger sequencing method. Sequences generated were processed and analysed using appropriate bioinformatics software. Results: The ticks were morphologically identified as Amblyomma spp. PCR profiling of the genomic DNA samples revealed the presence of the Rickettsia pathogen in 42 (21%) of the ticks collected from both Provinces. Out of the genes profiled, 14 (7%) were positive for 17KDa, 42 (21%) for ompA and 32 (16%) were positive for ompB genes respectively. The nucleotide blast of the sequenced genomes showed high similarity, as high as 100% with other reference Rickettsia (R.) africae in the GenBank. The phylogenetic analysis of the sequences further validated them as R. africae with their characteristic clustering pattern with related reference sequences. Conclusions: There is an abundance of R. africae in Amblyomma ticks collected from cattle in the study areas. This has serious public health implications as individuals who accidentally get infested with the ticks could acquire R. africae. Hence, adequate precautions in terms of sensitization of farmers about the risk and mass mobilization drive to control the vectors in the areas are highly recommended to safeguard public health.

Phylogenic position and low genomic diversity of 'Candidatus Rickettsia kotlanii' inferred by complete genome sequences of two Japanese isolates.

Many Rickettsia species of the spotted fever group (SFG) cause tick-borne diseases known as "spotted fever". One of the candidate SFG Rickettsia species is 'Candidatus Rickettsia kotlanii', which was first detected in Haemaphysalis concinna in Hungary in 2006. However, its precise phylogenetic position in the SFG is not clear because only single-gene sequence-based phylogenetic analyses were performed using very limited genes. Here, we present the complete genome sequences of two Japanese 'Ca. R. kotlanii' isolates, which differed only by a 135-bp insertion/deletion (InDel). Using these genomes and publicly available whole genome sequences of other Rickettsia species, the precise phylogenetic position of 'Ca. R. kotlanii' in Rickettsia was determined to be in a clade of the SFG. The phylogenetic relationships and average nucleotide identity of 'Ca. R. kotlanii' relative to the other species indicated that 'Ca. R. kotlanii' is an independent taxon in the SFG. Notably, although the genomes of the two isolates were almost identical, the isolates were obtained from different tick species in different regions and years, suggesting extremely low genomic diversity in 'Ca. R. kotlanii'. While the genome of 'Ca. R. kotlanii' is the smallest in the transitional group and SFG Rickettsia sequenced to date, we identified genes uniquely present or absent in 'Ca. R. kotlanii', but most were apparently degraded. Therefore, analyses of differences at the sequence (single nucleotide polymorphisms and small InDels) or gene expression level will be required to understand the functional or physiological features unique to 'Ca. R. kotlanii'. This article is protected by copyright. All rights reserved.

The life cycle of Dermacentor nuttalli from the Qinghai-Tibetan Plateau under laboratory conditions and detection of spotted fever group Rickettsia spp.

Dermacentor nuttalli has been a focus of study because tick-borne pathogens have been widely identified in this tick from northern and southwestern China. The aim of this study was to characterize the life cycle of D. nuttalli under laboratory conditions and to detect spotted fever group (SFG) Rickettsia in the midgut and salivary glands of both field-collected and first laboratory generation adults. D. nuttalli ticks were collected in the field on the Qinghai-Tibetan Plateau from March to April 2021 and their life cycle was studied under laboratory conditions. Tick identify was molecularly confirmed, and SFG Rickettsia were detected in the midgut and salivary glands of males and females by PCR targeting different rickettsial genes. The results showed that the life cycle of D. nuttalli under laboratory conditions was completed in an average of 86.1 days. High positivity of Rickettsia spp. was detected in the midgut and salivary glands of both males (92.0%) and females (93.0%) of field-collected D. nuttalli ticks. However, a relatively lower positivity (4.0-6.0%) was detected in first laboratory generation adults. Furthermore, sequencing analysis showed that the Rickettsia sequences obtained in this study shared 98.6 to 100% nucleotide identity with Rickettsia slovaca and Rickettsia raoultii isolated from Dermacentor spp. in China. Phylogenetic analysis of Rickettsia spp. based on the gltA, ompA, ompB and sca4 genes revealed that the Rickettsia sequences obtained could be classified as belonging to R. slovaca and R. raoultii clades. This study described for the first time the life cycle of D. nuttalli from the Qinghai-Tibetan Plateau under laboratory conditions. Two species of SFG Rickettsia were detected in the midgut and salivary glands of males and females in both field-collected and first laboratory-generation adults of D. nuttalli. Our study provides new insights into pathogen detection in ticks in the Qinghai-Tibet Plateau, and the relationships among hosts, ticks, and pathogens.

Rickettsia amblyommatis in Ticks: A Review of Distribution, Pathogenicity, and Diversity

Rickettsia amblyommatis is a potentially pathogenic species of Rickettsia within the spotted fever group vectored by ticks. While many studies have been published on this species, there is debate over its pathogenicity and the inhibitory role it plays in diagnosing illnesses caused by other spotted fever group Rickettsia species. Many publications have recorded the high infection prevalence of R. amblyommatis in tick populations at a global scale. While this species is rather ubiquitous, questions remain over the epidemiological importance of this possible human pathogen. With tick-borne diseases on the rise, understanding the exact role that R. amblyommatis plays as a pathogen and inhibitor of infection relative to other tick-borne pathogens will help public health efforts. The goal of this review was to compile the known literature on R. amblyommatis, review what we know about its geographic distribution, tick vectors, and pathogenicity, assess relatedness between various international strains from ticks by phylogenetic analysis and draw conclusions regarding future research needed.

Mapping the global distribution of spotted fever group rickettsiae: a systematic review with modelling analysis

Emerging and re-emerging spotted fever group (SFG) rickettsioses are increasingly recognised worldwide as threats to public health, yet their global distribution and associated risk burden remain poorly understood. In this systematic review and modelling analysis, we mapped global distributions of all confirmed species of SFG rickettsiae (SFGR) detected in animals, vectors, and human beings, using data collected from the literature. We assessed ecological drivers for the distributions of 17 major SFGR species using machine learning algorithms, and mapped model-predicted risks. Between Jan 1, 1906, and March 31, 2021, we found reports of 48 confirmed SFGR species, with 66 133 human infections worldwide, with a large spatial variation across the continents. 198 vector species were detected to carry 47 of these Rickettsia spp. (146 ticks, 24 fleas, 15 mosquitoes, six mites, four lice, two keds, and one bug). Based on model-predicted global distributions of the 17 major SFGR species, we found five spatial clusters aggregated by ecological similarity in terms of environmental and ecoclimatic features. Rickettsia felis is the leading SFGR species to which 4·4 billion (95% CI 3·8-5·3 billion) people are at risk, followed by Rickettsia conorii (3·7 billion) and Rickettsia africae (3·6 billion). The wide spectrum of vectors is contributing substantially to the increasing incidence of SFGR infections among humans. Awareness, diagnosis, and surveillance of SFGR infections should be improved in the high-risk regions, especially in areas where human infections are underreported. National Key Research and Development Program of China.

Prevalence of Spotted Fever Group Rickettsia and Candidatus Lariskella in Multiple Tick Species from Guizhou Province, China.

Rickettsiales (Rickettsia spp., Ehrlichia spp., and Anaplasma spp., etc.) are generally recognized as potentially emerging tick-borne pathogens. However, some bacteria and areas in China remain uninvestigated. In this study, we collected 113 ticks from mammals in Guizhou Province, Southwest China, and screened for the Rickettsiales bacteria. Subsequently, two spotted fever group Rickettsia species and one Candidatus Lariskella sp. were detected and characterized. "Candidatus Rickettsia jingxinensis" was detected in Rhipicephalus microplus (1/1), Haemaphysalis flava (1/3, 33.33%), Haemaphysalis kitaokai (1/3), and Ixodes sinensis (4/101, 3.96%), whereas Rickettsia monacensis was positive in H. flava (1/3), H. kitaokai (2/3), and I. sinensis ticks (74/101, 73.27%). At least two variants/sub-genotypes were identified in the R. monacensis isolates, and the strikingly high prevalence of R. monacensis may suggest a risk of human infection. Unexpectedly, a Candidatus Lariskella sp. belonging to the family Candidatus Midichloriaceae was detected from Ixodes ovatus (1/4) and I. sinensis (10/101, 9.90%). The gltA and groEL gene sequences were successfully obtained, and they show the highest (74.63-74.89% and 73.31%) similarities to "Candidatus Midichloria mitochondrii", respectively. Herein, we name the species "Candidatus Lariskella guizhouensis". These may be the first recovered gltA and groEL sequences of the genus Candidatus Lariskella.

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.

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.

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.

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.

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.

Molecular detection of spotted fever group Rickettsia species from seropositive human blood samples

The Sri Lankan Journal of Infectious Diseases (SLJID) is an open access, peer-reviewed, biannual journal published by the Sri Lankan Society for Microbiology (SSM). The Journal considers articles from all professional disciplines involved in the field of infectious diseases. The Journal has been in publication since 2011, is included in the DOAJ since 2018 and is a member of COPE since 2021. The SLJID practices a double-blind peer review policy. From 2022, the SLJID publishes accepted manuscripts online immediately after copy-editing, enabling rapid dissemination of scientific knowledge. The published articles are subsequently compiled into two issues in April and October. The SLJID does not charge any article processing or publication fee.

Characterization of a novel transitional group Rickettsia species (Rickettsia tillamookensis sp. nov.) from the western black-legged tick, Ixodes pacificus.

A previously unrecognized Rickettsia species was isolated in 1976 from a pool of Ixodes pacificus ticks collected in 1967 from Tillamook County, Oregon, USA. The isolate produced low fever and mild scrotal oedema following intraperitoneal injection into male guinea pigs (Cavia porcellus). Subsequent serotyping characterized this isolate as distinct from recognized typhus and spotted fever group Rickettsia species; nonetheless, the isolate remained unevaluated by molecular techniques and was not identified to species level for the subsequent 30 years. Ixodes pacificus is the most frequently identified human-biting tick in the western United States, and as such, formal identification and characterization of this potentially pathogenic Rickettsia species is warranted. Whole-genome sequencing of the Tillamook isolate revealed a genome 1.43 Mbp in size with 32.4 mol% G+C content. Maximum-likelihood phylogeny of core proteins places it in the transitional group of Rickettsia basal to both Rickettsia felis and Rickettsia asembonensis. It is distinct from existing named species, with maximum average nucleotide identity of 95.1% to R. asembonensis and maximum digital DNA-DNA hybridization score similarity to R. felis at 80.1%. The closest similarity at the 16S rRNA gene (97.9%) and sca4 (97.5%/97.6% respectively) is to Candidatus 'Rickettsia senegalensis' and Rickettsia sp. cf9, both isolated from cat fleas (Ctenocephalides felis). We characterized growth at various temperatures and in multiple cell lines. The Tillamook isolate grows aerobically in Vero E6, RF/6A and DH82 cells, and growth is rapid at 28 °C and 32 °C. Using accepted genomic criteria, we propose the name Rickettsia tillamookensis sp. nov., with the type strain Tillamook 23. Strain Tillamook 23 is available from the Centers for Disease Control and Prevention Rickettsial Isolate Reference Collection (WDCM 1093), Atlanta, GA, USA (CRIRC accession number RTI001T) and the Collection de Souches de l'Unité des Rickettsies (WDCM 875), Marseille, France (CSUR accession number R5043). Using accepted genomic criteria, we propose the name Rickettsia tillamookensis sp. nov., with the type strain Tillamook 23 (=CRIRC RTI001=R5043).

Ticking on Pandora\u2019s box: a prospective case-control study into \u2018other\u2019 tick-borne diseases

BackgroundTick-borne pathogens other than Borrelia burgdorferi sensu lato – the causative agent of Lyme borreliosis – are common in Ixodes ricinus ticks. How often these pathogens cause human disease is unknown. In addition, diagnostic tools to identify such diseases are lacking or reserved to research laboratories. To elucidate their prevalence and disease burden, the study ‘Ticking on Pandora’s Box’ has been initiated, a collaborative effort between Amsterdam University Medical Center and the National Institute for Public Health and the Environment.MethodsThe study investigates how often the tick-borne pathogens Anaplasma phagocytophilum, Babesia species, Borrelia miyamotoi, Neoehrlichia mikurensis, spotted fever group Rickettsia species and/or tick-borne encephalitis virus cause an acute febrile illness after tick-bite. We aim to determine the impact and severity of these tick-borne diseases in the Netherlands by measuring their prevalence and describing their clinical picture and course of disease.The study is designed as a prospective case-control study. We aim to include 150 cases – individuals clinically suspected of a tick-borne disease – and 3 matched healthy control groups of 200 persons each. The controls consist respectively of a group of individuals with either a tick-bite without complaints, the general population and of healthy blood donors. During a one-year follow-up we will acquire blood, urine and skin biopsy samples and ticks at baseline, 4 and 12 weeks. Additionally, participants answer modified versions of validated questionnaires to assess self-reported symptoms, among which the SF-36, on a 3 monthly basis.DiscussionThis article describes the background and design of the study protocol of ‘Ticking on Pandora’s Box’. With our study we hope to provide insight into the prevalence, clinical presentation and disease burden of the tick-borne diseases anaplasmosis, babesiosis, B. miyamotoi disease, neoehrlichiosis, rickettsiosis and tick-borne encephalitis and to assist in test development as well as provide recommendations for national guidelines.Trial registrationNL9258 (retrospectively registered at Netherlands Trial Register, trialregister.nl in in February 2021).

Rickettsia spp. in rodent-attached ticks in Estonia and first evidence of spotted fever group Rickettsia species Candidatus Rickettsia uralica in Europe

BackgroundRickettsia spp. are human pathogens that cause a number of diseases and are transmitted by arthropods, such as ixodid ticks. Estonia is one of few regions where the distribution area of two medically important tick species, Ixodes persulcatus and I. ricinus, overlaps. The nidicolous rodent-associated Ixodestrianguliceps has also recently been shown to be present in Estonia. Although no data are available on human disease(s) caused by tick-borne Rickettsia spp. in Estonia, the presence of three Rickettsia species in non-nidicolous ticks has been previously reported. The aim of this study was to detect, identify and partially characterize Rickettsia species in nidicolous and non-nidicolous ticks attached to rodents in Estonia.ResultsLarvae and nymphs of I.ricinus (n = 1004), I. persulcatus (n = 75) and I.trianguliceps (n = 117), all removed from rodents and shrews caught in different parts of Estonia, were studied for the presence of Rickettsia spp. by nested PCR. Ticks were collected from 314 small animals of five species [Myodes glareolus (bank voles), Apodemus flavicollis (yellow necked mice), A.agrarius (striped field mice), Microtus subterranius (pine voles) and Sorex araneus (common shrews)]. Rickettsial DNA was detected in 8.7% (103/1186) of the studied ticks. In addition to identifying R.helvetica, which had been previously found in questing ticks, we report here the first time that the recently described I.trianguliceps-associated Candidatus Rickettsia uralica has been identified west of the Ural Mountains.Graphical

Beyond the IFA: Revisiting the ELISA as a More Sensitive, Objective, and Quantitative Evaluation of Spotted Fever Group Rickettsia Exposure.

Based on limited serological studies, at least 10% of the US population has been exposed to spotted fever group Rickettsia (SFGR) species. The immunofluorescence antibody assay (IFA) has been the gold standard for the serodiagnosis of rickettsial infections such as spotted fever rickettsiosis (SFR). However, the IFA is semi-quantitative and subjective, requiring a high level of expertise to interpret it correctly. Here, we developed an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of Rickettsia parkeri infection in the guinea pig. Our ELISA is an objective, quantitative, and high-throughput assay that shows greater sensitivity and resolution in observed titers than the IFA. We methodically optimized relevant parameters in sequence for optimal signal-to-noise ratio and low coefficient of variation% values. We used a guinea pig model as it is a part of our overall research efforts to understand the immunological and clinical response to SFGR species after tick transmission. Guinea pigs are a useful model to study SFR and show clinical signs of SFR, such as fever and eschars. We anticipate that this assay will be easily adapted to other hosts, including humans and other SFGR species.

Detection of Novel Spotted Fever Group Rickettsiae (Rickettsiales: Rickettsiaceae) in Ticks (Acari: Ixodidae) in Southwestern China.

Spotted fever group rickettsiae, mainly maintained and transmitted by ticks, are important etiological agents of (re)emerging zoonotic diseases worldwide. It is of great significance to investigate spotted fever group rickettsiae in ticks in different areas for the prevention and control of rickettsioses. In this study, a total of 305 ticks were collected from wild and domestic animals in Chongqing, Guizhou, Yunnan, and Guangxi provinces of southwestern China during 2017-2019 and examined for the presence of spotted fever group rickettsiae by PCR with primers targeting the partial gltA, ompA, rrs, and htrA genes. Results showed that two spotted fever group rickettsiae species, including the pathogenic Candidatus Rickettsia jingxinensis (Rickettsiales: Rickettsiaceae) and a potential novel species Rickettsia sp. sw (Rickettsiales: Rickettsiaceae), were identified. The Ca. R. jingxinensis sequences were recovered from Rhipicephalus microplus (Ixodida: Ixodidae) and Haemaphysalis longicornis (Ixodida: Ixodidae) ticks and phylogenetically clustered with previous Ca. R. jingxinensis, Ca. R. longicornii (Rickettsiales: Rickettsiaceae), and Rickettsia sp. XY118 (Rickettsiales: Rickettsiaceae) strains. Rickettsia sp. sw was detected in Amblyomma geoemydae (Ixodida: Ixodidae) and Rh. microplus. Interestingly, as far as we know, this was the first report of Rickettsia (Rickettsiales: Rickettsiaceae) in A. geoemydae. Phylogenetic analyses indicated that this potential novel species was closely related to R. aeschlimannii (Rickettsiales: Rickettsiaceae) with gltA and ompA genes and grouped in a cluster composed of R. montanensis (Rickettsiales: Rickettsiaceae), R. raoultii (Rickettsiales: Rickettsiaceae), R. aeschlimannii, R. massiliae (Rickettsiales: Rickettsiaceae), and R. rhipicephali (Rickettsiales: Rickettsiaceae) with htrA, while formed a separate clade with rrs. The pathogenicity of Rickettsia sp. sw should be further confirmed. These results expand the knowledge of the geographical distribution and vector distribution of spotted fever group rickettsiae in China and are useful for assessing the potential public health risk.

Molecular Evidence of Novel Spotted Fever Group Rickettsia Species in Amblyomma albolimbatum Ticks from the Shingleback Skink (Tiliqua rugosa) in Southern Western Australia.

Tick-borne infectious diseases caused by obligate intracellular bacteria of the genus Rickettsia are a growing global problem to human and animal health. Surveillance of these pathogens at the wildlife interface is critical to informing public health strategies to limit their impact. In Australia, reptile-associated ticks such as Bothriocroton hydrosauri are the reservoirs for Rickettsia honei, the causative agent of Flinders Island spotted fever. In an effort to gain further insight into the potential for reptile-associated ticks to act as reservoirs for rickettsial infection, Rickettsia-specific PCR screening was performed on 64 Ambylomma albolimbatum ticks taken from shingleback skinks (Tiliqua rugosa) located in southern Western Australia. PCR screening revealed 92% positivity for rickettsial DNA. PCR amplification and sequencing of phylogenetically informative rickettsial genes (ompA, ompB, gltA, sca4, and 17kda) suggested that the single rickettsial genotype detected represented a novel rickettsial species, genetically distinct from but closely related to Rickettsia gravesii and within the rickettsia spotted fever group (SFG). On the basis of this study and previous investigations, it would appear that Rickettsia spp. are endemic to reptile-associated tick species in Australia, with geographically distinct populations of the same tick species harboring genetically distinct SFG Rickettsia species. Further molecular epidemiology studies are required to understand the relationship between these diverse Rickettsiae and their tick hosts and the risk that they may pose to human and animal health.

Identification and Characterization of "Candidatus Rickettsia Thierseensis", a Novel Spotted Fever Group Rickettsia Species Detected in Austria.

Rickettsia spp. are the second most common pathogens detected in Ixodes ricinus ticks in Austria after Borrelia burgdorferi sensu lato. Species belonging to the spotted fever group (SFG) are the causative agents for tick-borne rickettsiosis across the world. So far, only four SFG Rickettsia spp. were detected in Austria, namely R. helvetica, R. raoultii, R. monacensis and R. slovaca. Here, we describe the identification of a new SFG Rickettsia species detected in an I. ricinus tick. Sequencing of various rickettsial genes revealed a nucleotide sequence similarity of 99.6%, 98.5%, 97.3% and 98.5% to the gltA, ompA, ompB, and sca4 genes, respectively, of known and validated species. Additionally, sequencing of the htrA gene and 23S-5S intergenic spacer region also only showed 99.6% and 99.2%, respectively, similarity to known species. Therefore, and in accordance with current criteria for Rickettsia species discrimination, we hereby describe a new species of the SFG with putative pathogenic potential. We propose the name “Candidatus Rickettsia thierseensis” based on the village Thiersee in the Austrian province of Tyrol, where the carrying tick was found.

Molecular epidemiology of Rickettsia sp. and Coxiella burnetii collected from Hyalomma asiaticum in Bactrian camels (Camelus bactrianus) in inner Mongolia of China

The purpose of the present study was to determine the presence of Rickettsia sp. and Coxiella burnetii in Hyalomma asiaticum collected from Bactrian camels (Camelus bactrianus) in Inner Mongolia of China. A total of 385 H. asiaticum adults collected from Bactrian camels from Alxa in Inner Mongolia from 2017 to 2018 were examined using PCR combined with sequencing. The results indicated that 18 (4.7 %) and 5 (1.3 %) samples tested positive for Rickettsia and C. burnetii, respectively. The corresponding nucleotide identities among detected genes ranged between 99.7 % and 100 % for the sequences of 17 kD and ompB from Rickettsia and 16S rRNA and FtsX from C. burnetii. Two representative strains—Rickettsia IMA-1 strain and C. burnetii IMA-1 strain—were used for subsequent analysis. NCBI BLAST and phylogenetic analyses demonstrated that the detected strain Rickettsia IMA-1 may represent a novel species of spotted fever group (SFG) rickettsiae. The C. burnetii IMA-1 strain was grouped with Coxiella burnetii str. RSA439 (GenBank: CP040059.1). In addition, the Rickettsia sp. was successfully isolated from the ticks in Vero cells incubated at 28 °C. These findings indicate that the H. asiaticum collected off Bactrian camels in Inner Mongolia carried SFG Rickettsia species and C. burnetii and could contribute to the etiology of febrile illness in animals and humans.