Candidatus Rickettsia Tarasevichiae Research Articles

A “One health” approach to the understanding of mammals, ticks and Rickettsia interactions in the Andes of Colombia

Infectious diseases, especially zoonotic, represent a significant global threat to both human and animal health. Ticks are among the primary vectors of pathogens affecting wild and domestic animals, some of which can also cause severe human diseases. To effectively face zoonotic diseases, the "One Health" approach is being promoted to integrate the health of human, animals, and ecosystems. Here, we identify the associations between ticks, rickettsiae, wild and domestic mammals, and humans in the Andean region of Colombia. A total of 366 ticks of 17 species belonging to the genera Amblyomma, Dermacentor, Ixodes, Ornithodoros, and Rhipicephalus were collected as free-living organisms, or parasitizing humans, wild (22 species) and domestic (3 species) mammals. Infection with Rickettsia parkeri strain NOD, Rickettsia cf. monacensis and ‘Candidatus Rickettsia tarasevichiae’ was detected in 3.4% of the ticks analyzed (n = 3). This study highlights the diversity of ticks in humans and wild and domestic mammals in Colombia. It also underscores the risk these ectoparasites represent to human and animal health due to the potential transmission of zoonotic pathogens such as Rickettsia spp.

Demographic and zoological drivers of infectome diversity in companion cats with ascites.

Frequent studies reported the risks of cats as an intermediate host of zoonotic pathogens (e.g., SARS-CoV-2). Cats have a physically close interaction with their owners through activities like petting, kissing, and being licked on the cheek and hands. However, there are still limited studies that systematically investigate the infectome structure of cats. In this study, we employed a meta-transcriptomics approach to characterize 15 species of pathogens in cats, with Candidatus Rickettsia tarasevichiae first characterizing infection in diseased cats. Most feline diseases were better explained by the presence of virus-bacteria or virus-virus coinfection. The increase in infectome diversity could be influenced by a variety of predictors including age growth, temperature increase, and a higher proportion of females. A higher diversity of pathogens was harbored in young cats rather than adults. Importantly, we showed the value of linking the modern influx of meta-transcriptomics with comparative ecology and demography and of utilizing it to affirm that ecological and demographic variations impact the total infectome.

First detection of Candidatus Rickettsia tarasevichiae in Hyalomma marginatum ticks.

Ticks are important vectors of zoonotic diseases and play a major role in the circulation and transmission of many rickettsial species. The aim of this study was to investigate the carriage of Candidatus Rickettsia tarasevichiae (CRT) in a total of 1168 ticks collected in Inner Mongolia to elucidate the potential public health risk of this pathogen, provide a basis for infectious disease prevention, control and prediction and contribute diagnostic ideas for clinical diseases that present with fever in populations exposed to ticks. A total of four tick species, Haemaphysalis concinna (n = 21), Dermacentor nuttalli (n = 122), Hyalomma marginatum (n = 148), and Ixodes persulcatus (n = 877), were collected at nine sampling sites in Inner Mongolia, China, and identified by morphological and molecular biological methods. Reverse transcription PCR targeting the 16S ribosomal RNA (rrs), gltA, groEL, ompB and Sca4 genes was used to detect CRT DNA. Sequencing was used for pathogen species confirmation. The molecular epidemiological analysis showed that three species of ticks were infected with CRT, and the overall positive rate was as high as 42%. The positive rate of I. persulcatus collected in Hinggan League city was up to 96%, and that of I. persulcatus collected in Hulun Buir city was 50%. The pool positive rates of D. nuttalli and H. marginatum collected in Bayan Nur city and H. concinna collected in Hulun Buir city were 0%, 28% and 40%, respectively. This study revealed the high prevalence of CRT infection in ticks from Inner Mongolia and the first confirmation of CRT detected in H. marginatum in China. The wide host range and high infection rate in Inner Mongolia may dramatically increase the exposure of CRT to humans and other vertebrates. The role of H. marginatum in the transmission of rickettsiosis and its potential risk to public health should be further considered.

Genetic heterogeneity of <i>Rickettsia helvetica</i> population

Background. To date, the genetic variability of Rickettsia helvetica has not been sufficiently studied.The aim. To study the prevalence and genetic variability of R. helvetica in Ixodes spp. collected in Western Siberia and the Russian Far East.Materials and methods. Ixodes spp. collected from rodents in the Omsk province, Western Siberia (n = 280) and collected by flagging on Putyatin and Russky Islands in Primorsky Krai, Russian Far East (n = 482) were analyzed for the presence of Rickettsia spp. All positive samples were genotyped for the gltA gene fragment. For a number of R. helvetica samples, fragments of the 16S rRNA, ompA, ompB, sca4, htrA, and groEL genes and 23S–5S intergenic spacer were additionally sequenced.Results. Four Rickettsia species (R. helvetica, “Candidatus Rickettsia tarasevichiae”, “Candidatus Rickettsia uralica”, and “Candidatus Rickettsia mendelii”) were found. Of them, R. helvetica was identified in 72.2 % of Ixodes apronophorus and 18.8 % of Ixodes trianguliceps from the Omsk province and in single Ixodes persulcatus from the Omsk province and Putyatin Island. This is the first finding of Rickettsia spp. in I. apronophorus. All known R. helvetica sequences from this study and the GenBank database belonged to four well supported monopheletic groups forming genetic lineages I–IV. Lineage I included European isolates from Ixodes ricinus, Western Siberian isolates from I. persulcatus, and some sequences from I. apronophorus. All R. helvetica sequences from I. trianguliceps from the Omsk province and I. persulcatus from the Komi Republic and one sequence from I. apronophorus were assigned to lineage II. Most sequences from I. apronophorus formed lineage III; all known R. helvetica sequences from I. persulcatus from the Far East formed genetic lineage IV.Conclusion. The genetic heterogeneity of R. helvetica population was first demonstrated. Known isolates of R. helvetica are reliably assigned to four genetic lineages, but not in all cases the association of different lineages with a specific tick species or specific territory was observed.

Molecular surveillance reveals a potential hotspot of tick-borne disease in Yakeshi City, Inner Mongolia

A molecular surveillance of tick-borne diseases was performed in Hulunbuir City, Inner Mongolia. A total of 149 ticks including three species (Ixodes persulcatus, Haemaphysalis concinna, and Dermacentor silvarum) were collected. As many as 11 tick-borne bacterial pathogens were identified in them. Some of them have high positive rates. For example, Candidatus Rickettsia tarasevichiae was detected with a high prevalence of 72.48%, while Candidatus Lariskella sp. was detected in 31.54% of ticks. For both Rickettsia raoultii and Anaplasma phagocytophilum, two distinct genotypes were identified based on their phylogenetic trees based on 16S rRNA, gltA, and groEL sequences. Remarkable genetic diversity was also observed for 16S and flaB genes of Borreliella garinii, an agent of Lyme disease. Rickettsia heilongjiangensis causing Far-Eastern spotted fever (2.68%, 4/149), Ehrlichia muris causing human ehrlichiosis (4.70%, 7/149), Borrelia miyamotoi causing relapsing fever (2.01%, 3/149), and Borreliella afzelii causing Lyme disease (2.01%, 3/149) were also detected. Additionally, a previously uncharacterized Anaplasma species closely related to Anaplasma ovis was identified. Herein we name it “Candidatus Anaplasma mongolica”. Based on these results, we propose that Yakeshi City might be a potential hotspot of tick-borne diseases.

Applying next generation sequencing to detect tick-pathogens in Dermacentor nuttalli, Ixodes persulcatus, and Hyalomma asiaticum collected from Mongolia

Ticks and tick-borne diseases represent major threats to the public health of the Mongolian population, of which an estimated 26% live a traditional nomadic pastoralist lifestyle that puts them at increased risk for exposure. Ticks were collected by dragging and removal from livestock in Khentii, Selenge, Tuv, and Umnugovi aimags (provinces) during March-May 2020. Using next-generation sequencing (NGS) with confirmatory PCR and DNA sequencing, we sought to characterize the microbial species present in Dermacentor nuttalli (n = 98), Hyalomma asiaticum (n = 38), and Ixodes persulcatus (n = 72) tick pools. Rickettsia spp. were detected in 90.4% of tick pools, with Khentii, Selenge, and Tuv tick pools all having 100% pool positivity. Coxiella spp. were detected at an overall pool positivity rate of 60%, while Francisella spp. were detected in 20% of pools and Borrelia spp. detected in 13% of pools. Additional confirmatory testing for Rickettsia-positive pools demonstrated Rickettsia raoultii (n = 105), Candidatus Rickettsia tarasevichiae (n = 65) and R. slovaca/R. sibirica (n = 2), as well as the first report of Candidatus Rickettsia jingxinensis (n = 1) in Mongolia. For Coxiella spp. reads, most samples were identified as a Coxiella endosymbiont (n = 117), although Coxiella burnetii was detected in eight pools collected in Umnugovi. Borrelia species that were identified include Borrelia burgdorferi sensu lato (n = 3), B. garinii (n = 2), B. miyamotoi (n = 16), and B. afzelii (n = 3). All Francisella spp. reads were identified as Francisella endosymbiont species. Our findings emphasize the utility of NGS to provide baseline data across multiple tick-borne pathogen groups, which in turn can be used to inform health policy, determine regions for expanded surveillance, and guide risk mitigation strategies.

Molecular Detection of Anaplasma, Ehrlichia and Rickettsia Pathogens in Ticks Collected from Humans in the Republic of Korea, 2021.

Tick-borne pathogens (TBPs), transmitted by the bites of ticks, are of great medical and veterinary importance. They include bacteria, viruses, and protozoan parasites. To provide fundamental data on the risk of tick contact and public health strategies, we aimed to perform a molecular investigation on four tick-borne bacterial pathogens in ticks collected from humans across the Republic of Korea (ROK) in 2021. In total, 117 ticks were collected, including Haemaphysalis longicornis (56.4%), Amblyomma testudinarium (26.5%), Ixodes nipponensis (8.5%), H. flava (5.1%), and I. persulcatus (0.9%). Among the ticks, 20.5% (24/117) contained tick-borne bacterial pathogens, with infection rates of 17.9% for Rickettsia (Candidatus Rickettsia jingxinensis, R. tamurae, R. monacensis, and Candidatus Rickettsia tarasevichiae), 2.5% for Anaplasma (A. phagocytophilum, A. capra, and A. bovis), and 0.9% for Ehrlichia (Ehrlichia sp.). Additionally, the co-detection rate for R. monacensis and A. phagocytophilum was 0.9%. To our knowledge, this is the first report of A. capra and A. bovis detection in ticks collected from humans in the ROK. This study contributes to the understanding of the potential risk of tick contact and provides fundamental data for establishing a public health strategy for tick-borne disease management in the ROK.

Novel Genetic Lineages of Rickettsia helvetica Associated with Ixodes apronophorus and Ixodes trianguliceps Ticks.

Ixodes apronophorus is an insufficiently studied nidicolous tick species. For the first time, the prevalence and genetic diversity of Rickettsia spp. in Ixodes apronophorus, Ixodes persulcatus, and Ixodes trianguliceps ticks from their sympatric habitats in Western Siberia were investigated. Rickettsia helvetica was first identified in I. apronophorus with a prevalence exceeding 60%. "Candidatus Rickettsia tarasevichiae" dominated in I. persulcatus, whereas I. trianguliceps were infected with "Candidatus Rickettsia uralica", R. helvetica, and "Ca. R. tarasevichiae". For larvae collected from small mammals, a strong association was observed between tick species and rickettsiae species/sequence variants, indicating that co-feeding transmission in studied habitats is absent or its impact is insignificant. Phylogenetic analysis of all available R. helvetica sequences demonstrated the presence of four distinct genetic lineages. Most sequences from I. apronophorus belong to the unique lineage III, and single sequences cluster into the lineage I alongside sequences from European I. ricinus and Siberian I. persulcatus. Rickettsia helvetica sequences from I. trianguliceps, along with sequences from I. persulcatus from northwestern Russia, form lineage II. Other known R. helvetica sequences from I. persulcatus from the Far East group into the lineage IV. The obtained results demonstrated the high genetic variability of R. helvetica.

New insights into the impact of microbiome on horizontal and vertical transmission of a tick-borne pathogen

BackgroundThe impact of host skin microbiome on horizontal transmission of tick-borne pathogens , and of pathogen associated transstadial and transovarial changes in tick microbiome are largely unknown, but are important to control increasingly emerging tick-borne diseases worldwide.MethodsFocusing on a rickettsiosis pathogen, Rickettsia raoultii, we used R. raoultii-positive and R. raoultii-negative Dermacentor spp. tick colonies to study the involvement of skin microbiota in cutaneous infection with rickettsiae in laboratory mice, and the function of the tick microbiome on maintenance of rickettsiae through all tick developmental stages (eggs, larvae, nymphs, adults) over two generations.ResultsWe observed changes in the skin bacteria community, such as Chlamydia, not only associated with rickettsial colonization but also with tick feeding on skin. The diversity of skin microbiome differed between paired tick-bitten and un-bitten sites. For vertical transmission, significant differences in the tick microbiota between pathogenic rickettsia-positive and -negative tick chorts was observed across all developmental stages at least over two generations, which appeared to be a common pattern not only for R. raoultii but also for another pathogenic species, Candidatus Rickettsia tarasevichiae. More importantly, bacterial differences were complemented by functional shifts primed for genetic information processing during blood feeding. Specifically, the differences in tick microbiome gene repertoire between pathogenic Rickettsia-positive and -negative progenies were enriched in pathways associated with metabolism and hormone signals during vertical transmission.ConclusionsWe demonstrate that host skin microbiome might be a new factor determining the transmission of rickettsial pathogens through ticks. While pathogenic rickettsiae infect vertebrate hosts during blood-feeding by the tick, they may also manipulate the maturation of the tick through changing the functional potential of its microbiota over the tick’s life stages. The findings here might spur the development of new-generation control methods for ticks and tick-borne pathogens.BrauQCozpnNTXK3Qm2qXLRVideo

Human pathogens in ticks removed from humans in Hebei, China

Ticks are the hosts or vectors of many human pathogens, including viruses, bacteria and protozoa, and can transmit these causative agents to humans when feeding on human bodies. In this study, 26 ticks removed from humans in Hebei, China were tested for the presence of human-pathogenic microorganisms by Polymerase Chain Reaction (PCR) or Reversed Transcript PCR (RT-PCR). As a result, 11 ticks tested positive for at least one human pathogen. Specifically, four validated human pathogens, including Rickettsia raoultii, Candidatus Rickettsia tarasevichiae, Babesia venatorum, and Borrelia garinii, as well as Anaplasma ovis with zoonotic potential, were identified in Ixodes persulcatus, Dermacentor silvarum and Haemaphysalis concinna. Importantly, this is the first report of Anaplasma and Babesia species pathogenic to humans in Hebei province. Moreover, the co-infections, including double infection and quadruple infection were observed. In addition, Candidatus R. principis with unknown pathogenicity was identified in one tick, which may be the same species as Candidatus R. hongyuanensis based on the nucleotide identity and phylogenetic analysis. Concluding, four validated tick-borne pathogens and one with zoonotic potential were identified in ticks parasitizing humans, suggesting the potential high public health risk in the local human population.

Mild Infections with Multiple Spotted Fever Grouped Rickettsia Species among Forest Rangers with Tick Bites

Objectives: Tick-borne rickettsiae are increasingly recognized to cause human infections; however, a complete clinical spectrum is lacking. Thus, surveillance study was conducted among forest rangers with tick bites to describe the clinical manifestations. Methods: One hundred fifty-nine blood samples were obtained from individuals bitten by ticks and 780 tick samples collected in the same endemic region were examined for the presence of Rickettsia. Serum samples were tested for IgM and IgG antibodies against R. heilongjiangensis. Results: Twenty-five (15.7%) individuals were shown to be infected with 5 Rickettsia species, including 14 Candidatus Rickettsia tarasevichiae (CRT), 8 R. raoultii, 1 R. felis, 1 R. heilongjiangensis, and 1 R. massiliae. Five individuals (1 CRT, 1 R. heilongjiangensis, and 3 R. raoultii) had mild illnesses; the other 20 individuals were asymptomatic. CRT was present in 38.4% (274/713) of I. persulcatus and 6.4% (3/47) of Hae. concinna. R. raoultii was demonstrated in 30.0% (6/20) of D. silvarum and 14.9% (7/17) of Hae. concinna. R. heilongjiangensis was detected in 9.5% (2/21) of D. silvarum and 0.3% (2/713) of I. persulcatus. Conclusions: The clinical manifestations of these rickettsioses were non-specific and differed from traditional features, thus supporting the necessity of wider investigations involving individuals with tick bites to develop an early differential diagnosis.

Multilocus genetic analysis indicates taxonomic status of “Candidatus Rickettsia mendelii” as a separate basal group

To date, the phylogeny of Rickettsia spp. from basal groups is based on the small number of identified species. Thus, the finding of “Candidatus Rickettsia mendelii” in 2016 is of great interest. In this study, “Ca. R. mendelii” was first identified in the Asian region in a new carrier, Ixodes pavlovskyi. “Candidatus R. mendelii”, along with “Candidatus Rickettsia tarasevichiae”, were found in Ixodes ticks collected on Russky Island (the Far East), where I. pavlovskyi coexists with I. persulcatus. To establish the taxonomic position of “Ca. R. mendelii”, a detailed genetic study was carried out. “Candidatus R. mendelii” was genotyped by five genetic fragments (16S rRNA, gltA, and ompB genes, groESL operon, and 23S–5S IGS region); among them, the ompB gene, groESL operon and 23S–5S IGS region were sequenced for the first time. In addition, “Ca. R. tarasevichiae” was genetically characterized by eight genetic loci (16S rRNA, gltA, ompA, ompB, sca4, htrA genes, groESL operon, and 23S–5S IGS region), of which the sca4 gene was first determined. Phylogenetic analysis indicated that regardless of analyzed genetic loci, “Ca. R. mendelii” formed a separate well-supported cluster on each phylogenetic tree. Phylogenetic analysis based on concatenated sequences of gltA, ompB, and groEL gene fragments (total length of 3191 bp) demonstrated that “Ca. R. mendelii”, like Rickettsia bellii, is a basal group of Rickettsia.

Molecular detection of “Candidatus Rickettsia tarasevichiae” by Loop-mediated Isothermal Amplification (LAMP) of the ompA gene

“Candidatus Rickettsia tarasevichiae” (CRT) is increasingly being recognized as a disease causative agent in China and poses a great challenge to public health. Rapid and accurate detection is indispensable for laboratory diagnosis of infection caused by CRT and its surveillance in ticks. In the present study, a novel DNA-based loop-mediated isothermal amplification (LAMP) assay targeting the ompA gene was developed for the detection of CRT in tick samples. A set of universal primers specific to CRT were designed using PrimerExplorer V5 software. The analytical sensitivity, evaluated using recombinant plasmids containing the ompA gene, reached up to 1 copy per reaction, greater than that of the PCR assay targeting the same gene. This LAMP assay specifically detected CRT and showed no cross-reaction with other species common in China within the genus Rickettsia. In addition, this newly developed LAMP assay presented high diagnostic sensitivity of CRT detection validated by known positive DNA samples from ticks and simulated clinical samples. The applicability of the LAMP assay was evaluated by screening CRT from ticks, and the result showed that CRT circulation in Weichang County, China, was confirmed. Our findings indicate that this LAMP method is sensitive and specific for the detection of CRT and may have a potential application in the detection of CRT infection in patients and ticks.

Pathogens in Ixodes persulcatus and Ixodes ricinus ticks (Acari, Ixodidae) in Karelia (Russia)

Ixodid ticks (Acarina, Ixodidae) are vectors of dangerous human infections. The main tick species that determine the epidemiological situation for tick-borne diseases in northern Europe are Ixodes ricinus and Ixodes persulcatus. In recent years, significant changes in the number and distribution of these species have been observed, accompanied by an expansion of the sympatric range. This work summarizes the data of long-term studies carried out in Karelia since 2007 on the infection of I. persulcatus and I. ricinus ticks with various pathogens, including new viruses with unclear pathogenic potential. As a result, tick-borne encephalitis virus (TBEV, Siberian genotype), Alongshan virus, several representatives of the family Phenuiviridae, Borrelia afzelii, Borrelia garinii, Ehrlichia muris, Candidatus Rickettsia tarasevichiae and Candidatus Lariskella arthropodarum were identified. Data were obtained on the geographical and temporal variability of tick infection rates with these main pathogens. The average infection rates of I. persulcatus with TBEV and Borrelia burgdorferi sensu lato were 4.4% and 23.4% and those of I. ricinus were 1.1% and 11.9%, respectively. We did not find a correlation between the infection rate of ticks with TBEV, B. burgdorferi s.l. and Ehrlichia muris/chaffeensis with the sex of the vector. In general, the peculiarities of the epidemiological situation in Karelia are determined by the wide distribution and high abundance of I. persulcatus ticks and by their relatively high infection rate with TBEV and B. burgdorferi s.l. in most of the territory, including the periphery of the range.

Occurrence and clinical manifestations of tick-borne rickettsioses in Western Siberia: First Russian cases of Rickettsia aeschlimannii and Rickettsia slovaca infections

Rickettsia sibirica subsp. sibirica is a main agent of tick-borne rickettsioses in Western Siberia, Russia. Recently, the first cases of Rickettsia raoultii infection in patients hospitalized in Novosibirsk Province were described. The aim of this study was to establish the etiologic agents of tick-borne rickettsioses in Western Siberia during three epidemiological seasons.Clinical samples from 1008 patients hospitalized after tick bites in April–September 2017–2019 were examined by nested PCR for the presence of Rickettsia spp. All positive specimens were genetically characterized by sequencing of the gltA gene; some specimens were also genotyped based on the 16S rRNA, ompA, and ompB genes. Rickettsia spp. DNA was detected in clinical samples from 56 (5.6%) patients. Five Rickettsia species were identified: R. sibirica (n = 28), R. raoultii (n = 15), Rickettsia aeschlimannii (n = 3), “Candidatus Rickettsia tarasevichiae” (n = 2), and Rickettsia slovaca (n = 1). In addition, new unclassified Rickettsia genovariants were found in specimens from seven patients. Patients with R. raoultii infection presented rash, eschar, and high serum aminotransferase levels less frequently compared to patients with R. sibirica infections, but more frequently showed neurological symptoms. Among other patients, only persons with an R. aeschlimannii infection had rash and/or eschar, which are typical for tick-borne rickettsioses. The current study showed that R. raoultii is a common agent of tick-borne rickettsioses in Novosibirsk Province. DNA from R. aeschlimannii and R. slovaca was found in clinical samples of patients in the Russian Federation for the first time.

Survey of tick species and molecular detection of selected tick-borne pathogens in Yanbian, China.

Ticks and tick-borne diseases pose a significant threat to public health. In this study, we aimed to determine the tick species distribution and pathogens carried by ticks in Yanbian, China. A total of 2673 questing ticks were collected from eight counties and cities in Yanbian and were morphologically identified. The presence of Candidatus Rickettsia tarasevichiae (CRT), spotted fever group Rickettsia (SFGR), severe fever thrombocytopenia syndrome virus (SFTSV), Theileria, and other pathogens was confirmed using polymerase chain reaction (PCR) and real-time quantitative PCR assays, followed by phylogenetic and genotypic analyses. According to the morphological identification, the tick species in Yanbian consisted of Haemaphysalis longicornis, Ixodes persulcatus, Dermacentor silvarum, H. japonica, and H. concinna. In H. longicornis, CRT, SFGR, SFTSV and Theileria orientalis were detected, while CRT, SFGR, and SFTSV were detected in I. persulcatus, H. japonica, and D. silvarum. Only SFTSV was detected in H. concinna. Mixed infection with CRT and SFTSV was observed in I. persulcatus and H. japonica. The gene sequences of all tested pathogens exhibited 95.7%–100% identity with the corresponding sequences deposited in GenBank. Phylogenetic analysis showed that different SFGR and SFTSV genotypes were closely related to the Korean strains. This study is the first to describe the genetic diversity of SFGR Candidatus Rickettsia longicornii in H. longicornis in Yanbian, China, using the ompA, ompB, sca4, and rrs genes. These results provide epidemiological data to support the prevention and control of ticks and tick-borne diseases in the border areas of China, North Korea, and Russia.

Clinical and laboratory characteristics of tick-borne rickettsiosis related to <i>Rickettsia sibirica</i> and <i>Candidatus Rickettsia tarasevichiae</i>

The Siberian tick-borne typhus (STT) is the most common tick-borne rickettsiosis (TBR) in Russia, registered in 17 administrative territories of the Southern Siberia and the Far East. The aim of this study was to describe clinical picture, pathological data and results of laboratory diagnostics during fatal mixed infection caused by two Rickettsia species most common in Russia — pathogenic Rickettsia sibirica and poorly examined Candidatus R. tarasevichiae, including identification of its etiological agents. A four-year-old girl in Krasnoyarsk Krai in a hyperendemic focus of the Siberian tick-borne typhus after tick sucking revealed typical TBR symptoms (scab at the site of tick suction, fever, spotted rash, myalgia) and meningeal syndrome, which is not typical for the Siberian tick-borne typhus. The child died on the seventh day of illness. Autopsy data (hepatosplenomegaly; cerebral edema, which was the immediate cause of death) and the results of histological examination (productive vasculitis of the brain, spinal cord and skin, polymorphic cell perivascular infiltrates in the liver and lungs, serous meningitis, myeloid hyperplasia of the spleen and lymph nodes, interstitial lymphoid infiltration in the myocardium) confirmed the clinical diagnosis of tick-borne rickettsiosis. The patient’s blood and brain samples were tested for a wide range of tick-borne pathogens and enteric viruses that cause brain damage using PCR followed by sequencing of the positive samples. The DNA of Rickettsia sibirica and Candidatus Rickettsia tarasevichiae was found in both blood and brain samples. R. sibirica was identified by the nucleotide sequences of gene fragments gltA, ompA and ompB, and Candidatus R. tarasevichiae — by the gltA and ompB genes using nested PCR and sequencing. All amplified fragments were sequenced in both directions; the obtained sequences were deposited in the GenBank database under the inventory numbers MK048467–MK048475. We have not identified other tick-borne pathogens or intestinal viruses in the patient samples able to result in meningeal syndrome. The area where the child was sucked by the tick belongs to the regions with a high incidence of STT. Several species of mites coexist in this area; of these, Haemaphisalis concinna, the carrier of R. sibirica, dominates the populations of Ixodes mites, while Ixodes persulcatus, the main reservoir of Candidatus R. tarasevichiae, is less common. As a result of the conducted studies, for the first time in the Russian Federation, a verified case of a lethal infection associated with two species of Rickettsia — Rickettsia sibirica and Candidatus R. tarasevichiae — was identified and described.

Molecular detection and genetic diversity of Rickettsia spp. in pet dogs and their infesting ticks in Harbin, northeastern China

BackgroundPet dogs are important companion animals that share the environment within households, and play an important role in local community life. In addition, pet dogs also are reservoirs of zoonotic agents, including Rickettsia spp., thus increasing the risk of rickettsial infections in humans. It’s meaningful to investigate the epidemiology of rickettsial agents in pet dogs, and make contribute to the surveillance of rickettsioses in human in China.ResultsIn this study, a total of 496 pet dogs’ blood samples and 343 ticks infested in pet dogs were collected, and the presence and prevalence of Rickettsia were determined by amplifying the partial gltA and 17-kDa genes, with an overall positive rate of 8.1 % in blood samples and 14.0 % in tick samples. In addition, the rrs, gltA, groEL, and ompA genes of rickettsial were also recovered to determine the species of Rickettsia detected furtherly. Sequencing blast and phylogenetic analyses revealed the presence of three human pathogenic Rickettsia species (Rickettsia raoultii, Candidatus Rickettsia tarasevichiae and Rickettsia felis) in samples associated with pet dogs. Moreover, all the sequences of Rickettsia that we obtained presented close relationship with others available in GenBank, and Rickettsia raoultii was the most predominant Rickettsia species infected in pet dogs’ blood samples or in tick samples.ConclusionsThis study provides the molecular epidemiology data about the Rickettsia spp. infection associated with pet dogs in urban areas of Harbin city. Three rickettisae species pathogenic to humans were identified from pet dogs’ blood and the infested ticks in urban areas of Harbin city. Considering the intimate relationship between human and pets, these results indicate the potential transmission risk of human rickettisal infections from pet dogs through ectoparasites, and also highlighting that more attention should be paid to rickettsial infection in pet dogs and the infested ticks from the “One health” perspective.

Diversity of rickettsiae in domestic, synanthropic, and sylvatic mammals and their ectoparasites in a spotted fever-epidemic region at the western US-Mexico border.

Over one hundred cases of human rickettsiosis, many fatal, are reported annually across the US-Mexico transboundary region, representing a likely undercount. Although cases are often attributed to Rickettsia rickettsii, the agent of Rocky Mountain spotted fever, multiple other Rickettsia pathogens are present in North America. We conducted multiple-host surveillance of domestic, synanthropic, and sylvatic mammals and their ectoparasites to investigate the ecology of Rickettsia species in this region. A total of 499 mammals, including 83 dogs, 23 wild carnivores, five lagomorphs, and 388 rodents were sampled, and 413 fleas and 447 ticks belonging to 15 and 4 species, respectively, were collected during 2017 and 2018. We detected Rickettsia spp. DNA in one blood sample of coyote (Canis latrans), 11 ear tissues of rodents (10.6%), and 79 ectoparasites (9.5%). Of the 64 Rickettsia-positive fleas, 54 were Echidnophaga gallinacea and 10 were Pulex simulans, while of the 15 ticks, 11 were Rhipicephalus sanguineus s.l. and four Ixodes pacificus. The DNA sequence alignment of gltA and ompB regions revealed one and ten genetic variants of Rickettsia spp., respectively. These variants were clustered in clades of zoonotic species (R. felis, R. massiliae, R. parkeri, R. rickettsii, and R. typhi) and organisms of unknown pathogenic significance (R. asembonensis and Candidatus Rickettsia tarasevichiae). The finding of a coyote infected with R. rickettsii and the multiple zoonotic SFG rickettsial agents in the study area suggest that: 1) wild canids could serve as an amplifying host for RMSF, an alternate host for Rh. sanguineus s.l. ticks, and a means to spread infection and ticks over large areas; and 2) at least some of the human rickettsiosis cases attributed to R. rickettsii could be caused by other Rickettsia species. This study strongly supports the importance of multiple-host and vector eco-epidemiological studies and the One Health approach to better understand disease in a RMSF-epidemic region.

Prevalence of Multiple Tick-Borne Pathogens in Various Tick Vectors in Northeastern China.

Background: Tick-borne bacteria and protozoa can cause a variety of human and animal diseases in China. It is of great importance to monitor the prevalence and dynamic variation of these pathogens in ticks in ever-changing natural and social environment. Materials and Methods: Ticks were collected from Heilongjiang and Jilin provinces of northeastern China during 2018-2019 followed by morphological identification. The presence of Rickettsia spp., Anaplasma spp., Ehrlichia spp., Borrelia spp., Babesia spp., and Theileria spp. was examined by PCR and Sanger sequencing. The obtained sequences were subjected to phylogenetic analysis through Mega 7.0. Statistical analysis was performed using SPSS 24.0. Results: A total of 250 ticks from 5 species of 3 genera were collected. Ixodes and Haemaphysalis ticks carried more species of pathogens than Dermacentor, and the pathogens detected in Haemaphysalis japonica varied significantly among different sampling sites. The infection rates of Rickettsia spp., Anaplasma spp., Ehrlichia spp., Borrelia spp., Babesia spp., and Theileria spp. were 41.2%, 0, 2.0%, 7.2%, 1.2%, and 7.2%, respectively. Twelve pathogens were identified, among which Rickettsia raoultii (29.6%), Candidatus Rickettsia tarasevichiae (9.2%), and Theileria equi (4.4%) were the three most common ones. Rickettsia had its dominant vector, that is, R. raoultii had high infection rates in Dermacentor nuttalli and Dermacentor silvarum, Ca. R. tarasevichiae in Ixodes persulcatus, and Rickettsia heilongjiangensis in H. japonica. Interestingly, unclassified species were observed, including a Rickettsia sp., an Ehrlichia sp., a Borrelia sp., and a Babesia sp. Coinfections with different pathogens were identified in 9.2% of all tested ticks, with I. persulcatus most likely to be coinfected (23.8%) and Rickettsia spp. and Borrelia spp. as the most common combination (16.7%). Conclusions: The results of this study reflect high diversity and complexity of pathogens in ticks, which are useful for designing more targeted and effective control measures for tick-borne diseases in China.