Novel spotted fever group rickettsiae in Haemaphysalis qinghaiensis ticks from Gansu, Northwest China.
ABSTRACT: Rickettsia spp. are obligate intracellular bacteria and well known as transmitted by arthropods. These pathogens have a broad geographic distribution and a high degree of biological and clinical diversity. This study was conducted to determine the prevalence and molecular characterization of Rickettsia spp. in ticks collected from Gansu, where Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum were previously reported in ticks and ruminants.A total of 1,583 questing Haemaphysalis qinghaiensis ticks were collected and tested for the presence of Rickettsia spp. gltA gene by PCR. Samples positive for gltA were examined by specific primers targeted for the ompA gene of SFG rickettsiae. The infections were further validated by sequencing and positive samples were genetically characterized based on the gltA and ompA genes.In total, Rickettsia spp. infection was found in 179 (18.5 %) H. qinghaiensis tick pools by using PCR and primers specific for the gltA gene. Of those, 157 (16.3 %) tick pools were positive for SFG rickettsiae by PCR based on ompA gene. Amplification and molecular analysis of the nucleotide sequences of gltA and ompA genes indicated three potential novel spotted fever group rickettsiae in H. qinghaiensis ticks. These three potential novel spotted fever group rickettsiae were clustered together in a subgroup, which represents a sister taxon to and separates from other known four SFG rickettsiae subgroups.This study revealed a high infection rate of SFG rickettsiae in H. qinghaiensis ticks in northwest China. Three potential novel spotted fever group rickettsiae classified into a novel SFG rickettsiae subgroup were identified and named "Candidatus Rickettsia gannanii" related strains in recognition of the location where it was first detected.
Project description:Ixodid ticks were collected from vegetation and from humans, wild and domestic mammals in a rural area in the semi-arid Argentine Chaco in late spring 2006 to evaluate their potential role as vectors of Spotted Fever Group (SFG) rickettsiae. A total of 233 adult ticks, identified as Amblyomma parvum, Amblyomma tigrinum and Amblyomma pseudoconcolor, was examined for Rickettsia spp. We identified an SFG rickettsia of unknown pathogenicity, "Candidatus Rickettsia sp. strain Argentina", in A. parvum and A. pseudoconcolor by PCR assays targeting gltA, ompA, ompB and 17-kDa outer membrane antigen rickettsial genes. Rickettsia bellii was detected in a host-seeking male of A. tigrinum. Amblyomma parvum is widespread in the study area and is a potential threat to human health.
Project description:Spotted fever group (SFG) rickettsiae are obligate intracellular Gram-negative bacteria mainly associated with ticks. In Japan, several hundred cases of Japanese spotted fever, caused by Rickettsia japonica, are reported annually. Other Rickettsia species are also known to exist in ixodid ticks; however, their phylogenetic position and pathogenic potential are poorly understood. We conducted a nationwide cross-sectional survey on questing ticks to understand the overall diversity of SFG rickettsiae in Japan. Out of 2,189 individuals (19 tick species in 4 genera), 373 (17.0%) samples were positive for Rickettsia spp. as ascertained by real-time PCR amplification of the citrate synthase gene (gltA). Conventional PCR and sequencing analyses of gltA indicated the presence of 15 different genotypes of SFG rickettsiae. Based on the analysis of five additional genes, we characterised five Rickettsia species; R. asiatica, R. helvetica, R. monacensis (formerly reported as Rickettsia sp. In56 in Japan), R. tamurae, and Candidatus R. tarasevichiae and several unclassified SFG rickettsiae. We also found a strong association between rickettsial genotypes and their host tick species, while there was little association between rickettsial genotypes and their geographical origins. These observations suggested that most of the SFG rickettsiae have a limited host range and are maintained in certain tick species in the natural environment.
Project description:BACKGROUND: Detection of specific targets by PCR is used to confirm a diagnosis of spotted fever, but serological tests are still widely used. In this prospective study, nested PCR was performed on skin biopsy specimens to confirm the diagnosis of spotted fever. METHODS: In 58 clinically suspected cases of spotted fever, nested PCR, to detect gltA, 17 kDa lipoprotein antigen gene (17 kDa), ompA and ompB, from skin biopsy of the rash was performed. Sequencing was carried on amplicons representing the four targets to confirm specificity of amplification. This was followed by phylogenetic analysis using MEGA version 4.0 software. RESULTS: The gltA, 17 kDa, ompA, and ompB genes were detected from skin biopsy specimens in 38, 23, 27, and 22 individuals. Sequence analysis revealed that the gltA, 17 kDa, ompA, and ompB sequences belonged to spotted fever group (SFG) rickettsia. Of the six partial ompA gene sequences, only one was dissimilar to the previously reported 'Candidatus Rickettsia kellyi'. CONCLUSION: Further evidence indicates that SFG rickettsiae resembling 'Candidatus Rickettsia kellyi' cause fever and rash in southern India. More detailed phylogenetic analysis following isolation of rickettsia in culture is required for providing irrefutable proof for the occurrence of novel spotted fever rickettsiae in this region.
Project description:Evidence of spotted fever group (SFG) rickettsiae was obtained from flea pools and individual ticks collected at three sites in northwestern Peru within the focus of an outbreak of febrile disease in humans attributed, in part, to SFG rickettsia infections. Molecular identification of the etiologic agents from these samples was determined after partial sequencing of the 17-kDa common antigen gene (htrA) as well as pairwise nucleotide sequence homology with one or more of the following genes: gltA, ompA, and ompB. Amplification and sequencing of portions of the htrA and ompA genes in pooled samples (2 of 59) taken from fleas identified the pathogen Rickettsia felis. Four tick samples yielded molecular evidence of SFG rickettsiae. Fragments of the ompA (540-bp) and ompB (2,484-bp) genes were amplified from a single Amblyomma maculatum tick (tick 124) and an Ixodes boliviensis tick (tick 163). The phylogenetic relationships between the rickettsiae in these samples and other rickettsiae were determined after comparison of their ompB sequences by the neighbor-joining method. The dendrograms generated showed that the isolates exhibited close homology (97%) to R. aeschlimannii and R. rhipicephali. Significant bootstrap values supported clustering adjacent to this nodule of the SFG rickettsiae. While the agents identified in the flea and tick samples have not been linked to human cases in the area, these results demonstrate for the first time that at least two SFG rickettsia agents were circulating in northern Peru at the time of the outbreak. Furthermore, molecular analysis of sequences derived from the two separate species of hard ticks identified a possibly novel member of the SFG rickettsiae.
Project description:To obtain a better understanding of the current magnitude of tick-borne rickettsioses in Corsica, we used molecular methods to characterize the occurrence of Rickettsia spp. in ixodid ticks collected from domestic and wild animals. The presence of Rickettsia spp. was evaluated using real-time polymerase chain reaction targeting the gltA gene and by sequencing of gltA and ompA partial genes for species identification and phylogenetic analysis. Infection rates were calculated as the maximum-likelihood estimation (MLE) with 95% confidence intervals (CI). In total, 1117 ticks belonging to four genera (Rhipicephalus, Hyalomma, Ixodes, and Dermacentor) were collected from cattle, sheep, wild boars, and companion animals during July-August 2017 and July 2018-January 2019. Overall, Rickettsia DNA was detected in 208 of 349 pools of ticks (MLE = 25.6%, 95% CI: 22.6-28.8%). The molecular analysis revealed five different rickettsial species of the spotted-fever group (SFG). We highlighted the exclusive detection of Candidatus Ri. barbariae in R. bursa and of Ri. aeschlimanii in H. marginatum. Rickettsia slovaca was detected in D. marginatus collected from wild boars. This study provides the first evidence of the presence of Ri. monacensis in I. ricinus ticks isolated from a dog in Corsica. In conclusion, our data revealed wide dispersal of SFG Rickettsiae and their arthropod hosts in Corsica, highlighting the need for surveillance of the risk of infection for people living and/or working close to infected or infested animals.
Project description:Tick-borne infectious diseases caused by obligate intracellular bacteria of the genus <i>Rickettsia</i> 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 <i>Bothriocroton hydrosauri</i> are the reservoirs for <i>Rickettsia honei</i>, 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, <i>Rickettsia</i>-specific PCR screening was performed on 64 <i>Ambylomma albolimbatum</i> ticks taken from shingleback skinks (<i>Tiliqua rugosa</i>) located in southern Western Australia. PCR screening revealed 92% positivity for rickettsial DNA. PCR amplification and sequencing of phylogenetically informative rickettsial genes (<i>ompA, ompB, gltA, sca4,</i> and <i>17kda</i>) suggested that the single rickettsial genotype detected represented a novel rickettsial species, genetically distinct from but closely related to <i>Rickettsia gravesii</i> and within the rickettsia spotted fever group (SFG). On the basis of this study and previous investigations, it would appear that <i>Rickettsia</i> spp. are endemic to reptile-associated tick species in Australia, with geographically distinct populations of the same tick species harboring genetically distinct SFG <i>Rickettsia</i> species. Further molecular epidemiology studies are required to understand the relationship between these diverse <i>Rickettsiae</i> and their tick hosts and the risk that they may pose to human and animal health.
Project description:BACKGROUND:Rickettsia species belonging to the spotted fever group (SFG) cause infections in humans, domestic animals and wildlife. At least ten SFG Rickettsia species are known to occur in China. However, the distribution of rickettsiae in ticks and fleas in the border region of northwestern China have not been systematically studied to date. RESULTS:A total of 982 ticks (Rhipicephalus turanicus, Dermacentor marginatus, D. nuttalli and Haemaphysalis punctata) and 5052 fleas (18 flea species from 14 species of wild mammals) were collected in ten and five counties, respectively, of Xinjiang Uygur Autonomous Region (northwestern China). Tick and flea species were identified according to morphological and molecular characteristics. Seven sets of primers for amplifying the 17-kDa antigen gene (17-kDa), citrate synthase gene (gltA), 16S rRNA gene (rrs), outer membrane protein A and B genes (ompA, ompB), surface cell antigen 1 gene (sca1) and PS120-protein encoding gene (gene D) were used to identify the species of rickettsiae. Nine Rickettsia species have been detected, seven of them in ticks: R. aeschlimannii, R. conorii, R. raoultii, Rickettsia sibirica, R. slovaca, R. massiliae and "Candidatus R. barbariae". In addition, R. bellii and two genotypes of a rickettsia endosymbiont (phylogenetically in an ancestral position to R. bellii) have been detected from flea pools. CONCLUSIONS:This study provides molecular evidence for the occurrence of several SFG rickettsiae in Rhipicephalus turanicus, Dermacentor nuttalli and D. marginatus. Furthermore, R. bellii and two ancestral rickettsia endosymbionts are present in fleas infesting wild rodents in the border regions of northwestern China. These data extend our knowledge on the diversity of rickettsiae in Central Asia.
Project description:Tick-borne rickettsioses are caused by obligate intracellular bacteria belonging to the spotted fever group (SFG) rickettsiae. Although Spotted Fever is prevalent in the Middle East, no reports for the presence of tick-borne pathogens are available or any studies on the epidemiology of this disease in the West Bank. We aimed to identify the circulating hard tick vectors and genetically characterize SFG Rickettsia species in ixodid ticks from the West Bank-Palestinian territories.A total of 1,123 ixodid ticks belonging to eight species (Haemaphysalis parva, Haemaphysalis adleri, Rhipicephalus turanicus, Rhipicephalus sanguineus, Rhipicephalus bursa, Hyalomma dromedarii, Hyalomma aegyptium and Hyalomma impeltatum) were collected from goats, sheep, camels, dogs, a wolf, a horse and a tortoise in different localities throughout the West Bank during the period of January-April, 2014. A total of 867 ticks were screened for the presence of rickettsiae by PCR targeting a partial sequence of the ompA gene followed by sequence analysis. Two additional genes, 17 kDa and 16SrRNA were also targeted for further characterization of the detected Rickettsia species. Rickettsial DNA was detected in 148 out of the 867 (17%) tested ticks. The infection rates in Rh. turanicus, Rh. sanguineus, H. adleri, H. parva, H. dromedarii, and H. impeltatum ticks were 41.7, 11.6, 16.7, 16.2, 11.8 and 20%, respectively. None of the ticks, belonging to the species Rh. bursa and H. aegyptium, were infected. Four SFG rickettsiae were identified: Rickettsia massiliae, Rickettsia africae, Candidatus Rickettsia barbariae and Candidatus Rickettsia goldwasserii.The results of this study demonstrate the geographic distribution of SFG rickettsiae and clearly indicate the presence of at least four of them in collected ticks. Palestinian clinicians should be aware of emerging tick-borne diseases in the West Bank, particularly infections due to R. massiliae and R. africae.
Project description:Background: Ticks transmit a plethora of pathogens of zoonotic implications. Their distribution, diversity and the pathogens they transmit differ from one ecological location to another. Rickettsia africae is the agent of African tick bite fever found in South Africa, a zoonotic infection that is frequently reported among travelers who have visited many sub-Saharan African countries where the pathogen is prevalent. Methods: Ticks were collected from domestic animals in Raymond Nkandla Municipality, Eastern Cape, South Africa. The ticks were identified morphologically prior to DNA extraction followed by molecular identification of randomly selected ticks from the morphologically delineated groups. To assess for the presence of tick-borne pathogens belonging to Rickettsia spp. by PCR (polymerase chain reaction), we used specific primer pairs targeting the gltA, ompA and ompB genes. The selected amplified ticks, all positive ompB and forty three ompA amplicons were sequenced in a commercial sequencing facility. The obtained nucleotide sequences were edited and subjected to BLASTn for homology search and phylogenetic analyses were performed with MEGA 7 Version for genetic relationships with curated reference sequences in GenBank. Results: A total of 953 ticks collected in the study were delineated into three genera consisting of Amblyomma, Rhipicephalus and Hyalomma in decreasing order of abundance. The presence of rickettsial DNA was detected in 60/953 (6.3%) from the three genera of ticks screened. Genetic analyses of the DNA sequences obtained showed that they have phylogenetic relationship to members of the spotted fever group rickettsiae with R. africae, being the predominant SFGR (spotted fever group rickettsiae) detected in the screened ticks. Conclusion: This report shows that R. africae is the predominant spotted fever group rickettsiae in ticks collected from domestic animals in the study area and the human health impacts are not known.
Project description:Infection of dogs with Rickettsia spp. can result in inapparent, mild, or severe disease. Moreover, common dog ticks and fleas are able to transmit rickettsiae to nearby humans. In this study, the seroprevalence of spotted fever group (SFG) rickettsiae was determined in dogs of Costa Rica, as well as possible risk factors associated with exposure. An interview of owners and clinical examinations were performed in a country-wide sample of 441 dogs. IgG antibodies were determined in 399 dogs by indirect immunofluorescence assay (IFA) using antigens of Rickettsia rickettsii, R. amblyommatis, and R. felis. The presence of Rickettsia spp. gltA gene was evaluated by PCR in ticks and fleas. Poisson regression was performed to assess possible risk factors associated with seropositivity, as well as with having PCR-positive ticks and fleas. The overall seroprevalence to SFG rickettsiae was 10.0% (end titers 64 to 256). Rhipicephalus sanguineus s.l. (116/441; 26.3%) and Ctenocephalides felis (153/441; 34.7%) were the most common ectoparasites. Rickettsia DNA was detected in 30% (39/130) and 32.3% (56/173) of tick and flea pools, respectively. Seropositivity was significantly associated with mean age of 2 to 7?years, scrotal edema, walking problems, large size, and tick and flea infestation. Being a purebred dog was a possible protective factor. The presence of Rickettsia PCR-positive ticks was associated with being a purebred dog, while flea treatment was protective. Having PCR-positive fleas was associated with being purebred and the number of people in the dog's environment; protective factors were free roaming and being an outdoor dog. Results confirm that dogs in Costa Rica are exposed to different species of SFG rickettsiae. This may represent a risk to human health and underscores the need for accurate diagnosis in dogs and humans. Surveillance of rickettsial infection in canines may provide useful indicators to understand the epidemiology of these zoonoses.