Rickettsial DNA Research Articles

Detection of Rickettsial DNA in Ticksin Irkutsk Region

Individually 470 Dermacentor nuttalli and 46 Haemaphysalis concinna ticks collected in Irkutsk Region were analyzed using RT-PCR test systems. Rickettsia contamination of D. nuttalli was 82,3% including 1,3% of the ticks with R. sibirica DNA. H. concinna ticks were infected by rickettsia to 8,7% and the detected DNA in 6,5% of cases belonged to R. heilongjiangensis. PCR results were confirmed by phylogenetic analysis of the nucleotide sequences of three gene fragments: citrate synthase (gltA) and ompA and ompB surface proteins. Thus, circulation of two Rickettsia species pathogenic to humans was determined at the examined area.

Molecular detection of vector-borne pathogens in wild and domestic carnivores and their ticks at the human–wildlife interface

Urbanization of natural areas is considered one of the causes of the current apparent emergence of infectious diseases. Carnivores are among the species that adapt well to urban and periurban environments, facilitating cross-species disease transmission with domestic dogs and cats, and potentially with their owners. The prevalence of vector-borne pathogens (VBP) of zoonotic and veterinary interest was studied in sympatric wild and domestic carnivores into Barcelona Metropolitan Area (NE Spain). Blood or spleen samples from 130 animals, including 34 common genets (Genetta genetta), 12 red foxes (Vulpes vulpes), 10 stone martens (Martes foina), three Eurasian badgers (Meles meles), 34 free-roaming domestic cats and 37 dogs with outdoor access, were collected either in protected or adjacent residential areas. A total of 309 ticks (chiefly Rhipicephalus turanicus) were collected on these animals. The samples were analyzed with a battery of PCR assays targeting the DNA of Rickettsia spp., Anaplasmataceae, Coxiella burnetii, Bartonella spp., and Piroplasmida, and the amplicons were sequenced. The fox showed the highest prevalence (58%) and diversity of VBP (four pathogens), whereas none of the dogs were infected. Bartonella spp. (including B. clarridgeiae, B. henselae, and B. rochalimae) was the most prevalent pathogen. Infection of wild carnivores with Ehrlichia canis, C. burnetii, Theileria annae and Babesia vogeli was also confirmed, with some cases of coinfection observed. The presence of DNA of T. annae and B. vogeli was also confirmed in tick pools from four species of wild carnivores, supporting their role in piroplasmid life-cycle. By the sequencing of several target genes, DNA of Rickettsia massiliae was confirmed in 17 pools of Rh. turanicus, Rh. sanguineous, and Rh. pusillus from five different species, and Rickettsia conorii in one pool of Rh. sanguineous from a dog. None of the hosts from which these ticks were collected was infected by Rickettsia. Although carnivores may not be reservoir hosts for zoonotic Rickettsia, they can have an important role as mechanical dispersers of infected ticks.

Detection of a novel Rickettsia sp. in soft ticks (Acari: Argasidae) in Algeria

Argasid ticks are vectors of viral and bacterial agents that can infect humans and animals. In Africa, relapsing fever borreliae are neglected arthropod-borne pathogens that cause mild to deadly septicemia and miscarriage. It would be incredibly beneficial to be able to simultaneous detect and identify other pathogens transmitted by Argasid ticks. From 2012 to 2014, we conducted field surveys in 4 distinct areas of Algeria. We investigated the occurrence of soft ticks in rodent burrows and yellow-legged gull (Larus michahellis) nests in 10 study sites and collected 154 soft ticks. Molecular identification revealed the occurrence of two different soft tick genera and five species, including Carios capensis in yellow-legged gull nests and Ornithodoros occidentalis, Ornithodoros rupestris, Ornithodoros sonrai, Ornithodoros erraticus in rodent burrows. Rickettsial DNA was detected in 41/154, corresponding to a global detection rate of 26.6%. Sequences of the citrate synthase (gltA) gene suggest that this agent is a novel spotted fever group Rickettsia. For the first time in Algeria, we characterize a novel Rickettsia species by molecular means in soft ticks.

New evidence on tick-borne rickettsioses in the Altai region of Russia using primary lesions, serum and blood clots of molecular and serological study

Tick-borne rickettsioses (TBRs) have similar clinical symptoms and can give serological cross-reaction. We firstly found that in the natural foci of North Asian tick typhus (NATT) in the Altai region of Russia, TBRs can be caused by two Rickettsia species: Rickettsia sibirica subsp. sibirica (causative agent of NATT) and Rickettsia heilongjiangensis. Rickettsial DNA was detected in primary lesions, serum samples and blood clots using real-time PCR. Therefore, each case of TBRs should be verified by using molecular typing. TBR caused by R. sibirica subsp. sibirica – NATT dominates on the territory of Altai region.

High detection rate of Rickettsia africae in Amblyomma variegatum but low prevalence of anti-rickettsial antibodies in healthy pregnant women in Madagascar

Tick-borne spotted fever group (SFG) rickettsioses are emerging infectious diseases in Sub-Saharan Africa. In Madagascar, the endemicity of tick-borne rickettsiae and their vectors has been incompletely studied. The first part of the present study was conducted in 2011 and 2012 to identify potential anthropophilic tick vectors for SFG rickettsiae on cattle from seven Malagasy regions, and to detect and characterize rickettsiae in these ticks. Amblyomma variegatum was the only anthropophilic tick species found on 262 cattle. Using a novel ompB-specific qPCR, screening for rickettsial DNA was performed on 111 A. variegatum ticks. Rickettsial DNA was detected in 96 of 111 ticks studied (86.5%). Rickettsia africae was identified as the only infecting rickettsia using phylogenetic analysis of ompA and ompB gene sequences and three variable intergenic spacers from 11 ticks. The second part of the study was a cross-sectional survey for antibodies against SFG rickettsiae in plasma samples taken from healthy, pregnant women at six locations in Madagascar, two at sea level and four between 450 and 1300m altitude. An indirect fluorescent antibody test with Rickettsia conorii as surrogate SFG rickettsial antigen was used. We found R. conorii-seropositives at all altitudes with prevalences between 0.5% and 3.1%. Our results suggest that A. variegatum ticks highly infected with R. africae are the most prevalent cattle-associated tick vectors for SFG rickettsiosis in Madagascar. Transmission of SFG rickettsiosis to humans occurs at different altitudes in Madagascar and should be considered as a relevant cause of febrile diseases.

Amblyomma maculatum Feeding Augments Rickettsia parkeri Infection in a Rhesus Macaque Model: A Pilot Study

Rickettsia parkeri is an emerging eschar-causing human pathogen in the spotted fever group of Rickettsia and is transmitted by the Gulf coast tick, Amblyomma maculatum. Tick saliva has been shown to alter both the cellular and humoral components of the innate and adaptive immune systems. However, the effect of this immunomodulation on Rickettsia transmission and pathology in an immunocompetent vertebrate host has not been fully examined. We hypothesize that, by modifying the host immune response, tick feeding enhances infection and pathology of pathogenic spotted fever group Rickettsia sp. In order to assess this interaction in vivo, a pilot study was conducted using five rhesus macaques that were divided into three groups. One group was intradermally inoculated with low passage R. parkeri (Portsmouth strain) alone (n = 2) and another group was inoculated during infestation by adult, R. parkeri-free A. maculatum (n = 2). The final macaque was infested with ticks alone (tick feeding control group). Blood, lymph node and skin biopsies were collected at several time points post-inoculation/infestation to assess pathology and quantify rickettsial DNA. As opposed to the tick-only animal, all Rickettsia-inoculated macaques developed inflammatory leukograms, elevated C-reactive protein concentrations, and elevated TH1 (interferon-γ, interleukin-15) and acute phase inflammatory cytokines (interleukin-6) post-inoculation, with greater neutrophilia and interleukin-6 concentrations in the tick plus R. parkeri group. While eschars formed at all R. parkeri inoculation sites, larger and slower healing eschars were observed in the tick feeding plus R. parkeri group. Furthermore, dissemination of R. parkeri to draining lymph nodes early in infection and increased persistence at the inoculation site were observed in the tick plus R. parkeri group. This study indicates that rhesus macaques can be used to model R. parkeri rickettsiosis, and suggests that immunomodulatory factors introduced during tick feeding may enhance the pathogenicity of spotted fever group Rickettsia.

Genetic variability of Rickettsia spp. in Ixodes persulcatus/Ixodes trianguliceps sympatric areas from Western Siberia, Russia: Identification of a new Candidatus Rickettsia species

Rickettsia spp. are the causative agents of a number of diseases in humans. These bacteria are transmitted by arthropods, including ixodid ticks. DNA of several Rickettsia spp. was identified in Ixodes persulcatus ticks, however, the association of Ixodes trianguliceps ticks with Rickettsia spp. is unknown. In our study, blood samples of small mammals (n=108), unfed adult I. persulcatus ticks (n=136), and I. persulcatus (n=12) and I. trianguliceps (n=34) ticks feeding on voles were collected in two I. persulcatus/I. trianguliceps sympatric areas in Western Siberia. Using nested PCR, ticks and blood samples were studied for the presence of Rickettsia spp. Three distinct Rickettsia species were found in ticks, but no Rickettsia species were found in the blood of examined voles. Candidatus Rickettsia tarasevichiae DNA was detected in 89.7% of unfed I. persulcatus, 91.7% of engorged I. persulcatus and 14.7% of I. trianguliceps ticks. Rickettsia helvetica DNA was detected in 5.9% of I. trianguliceps ticks. In addition, a new Rickettsia genetic variant was found in 32.4% of I. trianguliceps ticks. Sequence analysis of the 16S rRNA, gltA, ompA, оmpB and sca4 genes was performed and, in accordance with genetic criteria, a new Rickettsia genetic variant was classified as a new Candidatus Rickettsia species. We propose to name this species Candidatus Rickettsia uralica, according to the territory where this species was initially identified. Candidatus Rickettsia uralica was found to belong to the spotted fever group. The data obtained in this study leads us to propose that Candidatus Rickettsia uralica is associated with I. trianguliceps ticks.

Spotted fever group rickettsiae detected in immature stages of ticks parasitizing on Iberian endemic lizard Lacerta schreiberi Bedriaga, 1878

Spotted fever rickettsioses are tick-borne diseases of growing public health concern. The prevalence of rickettsia-infected ticks and their ability to parasitize humans significantly influence the risk of human infection. Altogether 466 Ixodes ricinus ticks (428 nymphs and 38 larvae) collected from 73 Lacerta schreiberi lizards were examined by PCR targeting the citrate synthetase gene gltA for the presence of Rickettsia spp. Rickettsial DNA was detected in 47% of nymphs and 31.6% of larvae. They were subsequently subjected to a second PCR reaction using primers derived from the outer membrane protein rOmpA encoding gene (ompA) to detect spotted fever group rickettsiae (SFG). This analysis shows that 41.4% of nymphs and 7.9% of larvae collected from the lizards contain DNA of SFG rickettsiae. Sequencing of 43 randomly selected samples revealed two different haplotypes, both closely related to R. monacensis (39 and 4 samples, respectively). The remaining ompA negative Rickettsia spp. samples were determined to be R. helvetica based on sequencing of ompB and gltA fragments. Our results indicate that the role of Iberian endemic lizard L. schreiberi and its ectoparasites in the ecology and epidemiology of zoonotic SFG rickettsioses may be appreciable.

Detection and identification of Rickettsia species in Ixodes tick populations from Estonia

A total of 1640 ticks collected in different geographical parts of Estonia were screened for the presence of Rickettsia species DNA by real-time PCR. DNA of Rickettsia was detected in 83 out of 1640 questing ticks with an overall prevalence of 5.1%. The majority of the ticks infected by rickettsiae were Ixodes ricinus (74 of 83), while 9 of the 83 positive ticks were Ixodes persulcatus.For rickettsial species identification, a part of the citrate synthase gltA gene was sequenced. The majority of the positive samples were identified as Rickettsia helvetica (81 out of 83) and two of the samples were identified as Rickettsia monacensis and Candidatus R. tarasevichiae, respectively.Genetic characterization based on the partial gltA gene showed that the Estonian sequences within the R. helvetica, R. monacensis and Candidatus R. tarasevichiae species demonstrated 100% similarity with sequences deposited in GenBank, originating from Rickettsia species distributed over large territories from Europe to Asia.

Bacteria of the genus Rickettsia in ticks (Acari: Ixodidae) collected from birds in Costa Rica

The aim of this study was to document the presence of Rickettsia spp. in ticks parasitizing wild birds in Costa Rica. Birds were trapped at seven locations in Costa Rica during 2004, 2009, and 2010; then visually examined for the presence of ticks. Ticks were identified, and part of them was tested individually for the presence of Rickettsia spp. by polymerase chain reaction (PCR) using primers targeting fragments of the rickettsial genes gltA and ompA. PCR products were DNA-sequenced and analyzed in BLAST to determine similarities with previously reported rickettsial agents. A total of 1878 birds were examined, from which 163 birds (9%) were infested with 388 ticks of the genera Amblyomma and Ixodes. The following Amblyomma (in decreasing order of abundance) were found in immature stages (larvae and nymphs): Amblyomma longirostre, Amblyomma calcaratum, Amblyomma coelebs, Amblyomma sabanerae, Amblyomma varium, Amblyomma maculatum, and Amblyomma ovale. Ixodes ticks were represented by Ixodes minor and two unclassified species, designated here as Ixodes sp. genotype I, and Ixodes sp. genotype II. Twelve of 24 tested A. longirostre ticks were found to be infected with ‘Candidatus Rickettsia amblyommii’, and 2 of 4 A. sabanerae were found to be infected with Rickettsia bellii. Eight of 10 larval Ixodes minor were infected with an endosymbiont (a novel Rickettsia sp. agent) genetically related to the Ixodes scapularis endosymbiont. No rickettsial DNA was found in A. calcaratum, A. coelebs, A. maculatum, A. ovale, A. varium, Ixodes sp. I, and Ixodes sp. II. We report the occurrence of I. minor in Costa Rica for the first time and a number of new bird host–tick associations. Moreover, ‘Candidatus R. amblyommii’ and R. bellii were found in A. longirostre and A. sabanerae, respectively, in Costa Rica for the first time.

First detections of Rickettsia helvetica and R. monacensis in ectoparasitic mites (Laelapidae and Trombiculidae) infesting rodents in south-western Slovakia.

Information on circulation of rickettsiae between small mammals and their ectoparasitic mites is scarce. In this study, we investigated infestation rates of rodents with mites in some areas of SW Slovakia and the role of mites as possible vectors of pathogenic rickettsiae. A total of 615 rodents of five species were caught during 2010-2012. All individuals were examined for ectoparasites which resulted in 2821 mites belonging to three species of Laelapidae and six species of Trombiculidae. The most common Laelapidae species was Laelaps agilis (81.25%), followed by Haemogamasus nidi, and Eulaelaps stabularis. Hirsutiella zachvatkini (16.52%) was the dominant species of the family Trombiculidae. DNA extracted from rodent blood and ectoparasitic mites was examined for the presence of rickettsiae by PCR. By pooling mites, 345 pool samples were created, of which 112 (32.46%) were found to be positive for the rickettsial DNA. From 487 examined rodent blood samples, rickettsial DNA was found in 46 (9.44%). Sequencing DNA from the positive blood samples and mites revealed the identity of Rickettsia helvetica and Rickettsia monacensis. The results of the study suggest that ectoparasitic mites may be reservoirs as well as vectors of some pathogenic rickettsiae.

The detection of vector-borne-disease-related DNA in human stool paves the way to large epidemiological studies.

The detection of Plasmodium spp. by the molecular analysis of human feces was reported to be comparable to detection in the blood. We believe that for epidemiological studies using molecular tools, it would be simpler to use feces, which are easier to obtain and require no training for their collection. Our aim was to evaluate the usefulness of feces for the detection of these pathogens towards developing a new tool for their surveillance. Between 2008 and 2010, 451 human fecal samples were collected in two Senegalese villages in which malaria and rickettsioses are endemic. Rickettsia and Plasmodium DNA were detected using quantitative PCR targeting Rickettsia of the spotted fever group, R. felis and Plasmodium spp. Two different sequences were systematically targeted for each pathogen. Twenty of the 451 fecal samples (4.4 %) were positive for Rickettsia spp., including 8 for R. felis. Inhabitants of Dielmo were more affected (18/230, 7.8 %; p = 0.0008) compared to those of Ndiop (2/221, 0.9 %). Children under 15 years of age were more often positive (19/285, 6.7 %) than were older children (1/166, 0.6 %; p = 0.005, odds ratio = 11.79). Only one sample was positive for Plasmodium spp. This prevalence is similar to that found in the blood of the Senegalese population reported previously. This preliminary report provides a proof of concept for the use of feces for detecting human pathogens, including microorganisms that do not cause gastroenteritis, in epidemiological studies.

Molecular detection of the human pathogenic Rickettsia sp. strain Atlantic rainforest in Amblyomma dubitatum ticks from Argentina.

To date, three tick-borne pathogenic Rickettsia species have been reported in different regions of Argentina, namely, R. rickettsii, R. parkeri, and R. massiliae. However, there are no reports available for the presence of tick-borne pathogens from the northeastern region of Argentina. This study evaluated the infection with Rickettsia species of Amblyomma dubitatum ticks collected from vegetation and feeding from capybaras (Hydrochoerus hydrochaeris) in northeastern Argentina. From a total of 374 A. dubitatum ticks collected and evaluated by PCR for the presence of rickettsial DNA, 19 were positive for the presence of Rickettsia bellii DNA, two were positive for Rickettsia sp. strain COOPERI, and one was positive for the pathogenic Rickettsia sp. strain Atlantic rainforest. To our knowledge, this study is the first report of the presence of the human pathogen Rickettsia sp. strain Atlantic rainforest and Rickettsia sp. strain COOPERI in Argentina. Moreover, our findings posit A. dubitatum as a potential vector for this pathogenic strain of Rickettsia.

Serologic and Molecular Prevalence ofRickettsia helveticaandAnaplasma phagocytophilumin Wild Cervids and Domestic Mammals in the Central Parts of Sweden

Both Rickettsia helvetica and Anaplasma phagocytophilum are common in Ixodes ricinus ticks in Sweden. Knowledge is limited regarding different animal species' competence to act as reservoirs for these organism. For this reason, blood samples were collected from wild cervids (roe deer, moose) and domestic mammals (horse, cat, dog) in central Sweden, and sera were tested using immunofluorescence assay to detect antibodies against spotted fever rickettsiae using Rickettsia helvetica as antigen. Sera with a titer ≥1:64 were considered as positive, and 23.1% (104/450) of the animals scored positive. The prevalence of seropositivity was 21.5% (23/107) in roe deer, 23.3% (21/90) in moose, 36.5% (23/63) in horses, 22.1% (19/90) in cats, and 17.0% (17/100) in dogs. PCR analysis of 113 spleen samples from moose and sheep from the corresponding areas were all negative for rickettsial DNA. In roe deer, 85% (91/107) also tested seropositive for A. phagocytophilum with a titer cutoff of 1:128. The findings indicate that the surveyed animal species are commonly exposed to rickettsiae and roe deer also to A. phagocytophilum.

Evidence for a specific host-endosymbiont relationship between 'Rickettsia sp. genotype RF2125' and Ctenocephalides felis orientis infesting dogs in India.

BackgroundFleas of the genus Ctenocephalides serve as vectors for a number of rickettsial zoonoses, including Rickettsia felis. There are currently no published reports of the presence and distribution of R. felis in India, however, the ubiquitous distribution of its vector Ctenocephalides felis, makes it possible that the pathogen is endemic to the region. This study investigates the occurrence of Rickettsia spp. infection in various subspecies of C. felis infesting dogs from urban areas of Mumbai, Delhi and Rajasthan in India.MethodsIndividual fleas collected off 77 stray dogs from Mumbai, Delhi and Rajasthan were screened for Rickettsia spp. by a conventional PCR targeting the ompB gene. Further genetic characterisation of Rickettsia-positive fleas was carried out using nested PCR and phylogenetic analysis of partial DNA sequences of the gltA and ompA genes. Ctenocephalides spp. were morphologically and genetically identified by PCR targeting a fragment of cox1 gene.ResultsOverall, 56/77 fleas (72.7%), including 22/24 (91.7%) from Delhi, 32/44 (72.7%) from Mumbai and 2/9 (22.2%) from Rajasthan were positive for Rickettsia DNA at the ompB gene. Sequences of gltA fragments confirmed the amplification of Rickettsia sp. genotype RF2125. The ompA gene of Rickettsia sp. genotype RF2125 was characterised for the first time and shown 96% identical to R. felis. Three species of Ctenocephalides were identified, with the Ctenocephalides felis orientis being the dominant flea species (69/77; 89.6%) in India, followed by Ctenocephalides felis felis (8/77; 10.4%).ConclusionsHigh occurrence of Rickettsia sp. genotype RF2125 in C. felis orientis and the absence of R. felis suggests a specific vector-endosymbiont adaptation and coevolution of the Rickettsia felis-like sp. within subspecies of C. felis.

Clinical Presentation, Convalescence, and Relapse of Rocky Mountain Spotted Fever in Dogs Experimentally Infected via Tick Bite

Rocky Mountain spotted fever (RMSF) is a tick-borne disease caused by R. rickettsii in North and South America. Domestic dogs are susceptible to infection and canine RMSF can be fatal without appropriate treatment. Although clinical signs of R. rickettsii infection in dogs have been described, published reports usually include descriptions of either advanced clinical cases or experimental infections caused by needle-inoculation of cultured pathogen rather than by tick bite. The natural progression of a tick-borne R. rickettsii infection has not been studied in sufficient detail. Here, we provide a detailed description of clinical, hematological, molecular, and serological dynamics of RMSF in domestic dogs from the day of experimental exposure to infected ticks through recovery. Presented data indicate that neither the height/duration of fever nor detection of rickettsial DNA in dogs' blood by PCR are good indicators for clinical prognosis. Only the apex and subsequent subsidence of neutrophilia seem to mark the beginning of recovery and allow predicting a favorable outcome in Rickettsia-infected dogs, even despite the continuing persistence of mucosal petechiae and skin rash. On the other hand the appropriate (doxycycline) antibiotic therapy of sufficient duration is crucial in prevention of RMSF relapses in dogs.

Detection of Rickettsia felis in Wild Mammals from Three Municipalities in Yucatan, Mexico.

The aim of this study was to provide information of the occurrence of Rickettsia felis in wild mammals from three municipalities in Yucatan, Mexico. The reactivity of rodent serum to Rickettsia antigens was detected in 80.9% (17 of 21) samples using immunofluorescence assay. Polymerase chain reaction identified rickettsial DNA in spleens of 43.5% (10 of 23) rodents and 57.1% (4 of 7) opossums. The identification of the rickettsial DNA was confirmed as R. felis by restriction fragment length polymorphism and DNA sequencing. This study comprises the first report of R. felis detection in wild mammals in Yucatan.

Detection of rickettsial DNA in ticks and wild boars in Kyoto City, Japan.

ABSTRACTThe tick is a well-known vector for arthropod-borne pathogens, such as tick-borne encephalitis, Lyme disease, Japanese spotted fever and severe fever with thrombocytopenia syndrome. It is therefore important to know the tick population and distribution in our environment and wild animals in order to prevent tick-borne diseases. Here, we report the results of tick surveillance from May to September 2011 at 14 geographical points and in 5 wild boars in Kyoto City, Kyoto prefecture, Japan. We collected 3,198 ticks comprising 5 tick species, Haemaphysalis (H.) longicornis, H. flava, H. kitaokai, Amblyomma testudinarium and Dermacentor taiwanensis. Interestingly, the proportion of tick species varied according to geographical region within the city. The ticks collected in the city were reported as potential vectors of pathogens, such as rickettsiosis. We detected rickettsial DNA by PCR in 71.1% of 201 ticks investigated. The ticks that carried rickettsiae were distributed across the whole the city. The sequences of PCR-amplified DNA fragments were determined and showed similarities to spotted fever group rickettsiae. Although their pathogenicity for animals including humans is still unclear, it is important to stay alert and pay attention to tick-borne diseases in order to ensure the safety of the citizens of the city as well as that of visitors.

Ticks and rickettsial infection in the wildlife of two regions of the Brazilian Amazon.

During 2009-2012, wild animals and their ticks were sampled in two areas within the Amazon biome of Brazil, in the states of Mato Grosso and Pará. Animal tissues, blood, and ticks were molecularly tested for Rickettsia and Coxiella DNA. A total of 182 wild animals were sampled, comprising 28 mammalian, five avian, and three reptilian species. Animal tissues or blood were all negative for Rickettsia or Coxiella DNA. A total of 454 ticks (22 larvae, 226 nymphs, 127 males, 79 females) were collected from 52 (28.6%) animals, and identified into 15 species: Amblyomma cajennense, A. naponense, A. humerale, A. nodosum, A. goeldii, A. oblongoguttatum, A. longirostre, A. calcaratum, A. coelebs, A. pacae, A. geayii, A. rotundatum, A. auricularium, A. ovale, and Haemaphysalis juxtakochi. While no Coxiella DNA was identified in ticks, six Rickettsia species were detected in the ticks. "Candidatus Rickettsia amblyommii" was the most common agent, detected in four tick species, A. cajennense, A. auricularium, A. longirostre, and A. humerale. The second most common agent, R. bellii, was detected in A. humerale and A. naponense. Rickettsia rhipicephali was detected in H. juxtakochi, and R. felis in A. humerale. Two possible new Rickettsia species were detected in A. naponense ticks, namely, a novel spotted fever group agent close-related to R. africae in Pará, and a novel Canadensis group agent in Mato Grosso. Results of the present study expand our knowledge on the tick fauna, and on the yet infantile knowledge of tick-borne rickettsiae in the Amazon biome.

The role of cats in the eco-epidemiology of spotted fever group diseases.

BackgroundMediterranean Spotted Fever (MSF), whose etiological agent is R. conorii, is one of the oldest described vector-borne infectious diseases. Although it is endemic in the Mediterranean area, clinical cases have also been reported in other regions. R. massiliae-Bar29 is related to MSF cases. This strain is distributed worldwide. R. conorii and R. massiliae-Bar29 are transmitted by ticks. Dogs are considered the sentinel of R. conorii infection. Cats could also be involved in their transmission. Rickettsia felis, etiological agent of Flea-borne spotted fever, is mainly transmitted by the cat flea, Ctenocephalides felis. Up to now, the role of cats in its transmission is not entirely elucidated. The aim of the study is to analyze the infection in cats by these microorganisms.MethodsThe study was undertaken in Northeastern Spain. Twenty municipalities of seven regions participated in the study. 212 cats (pets and stray cats) were analyzed. Variables surveyed were: date of collection, age, sex, municipality, source, living place, outdoor activities, health status, type of disease, contact with other animals, and ectoparasite infestation. Sera were evaluated by indirect immunofluorescence antibody assay (IFA). Molecular detection (real-time PCR and sequencing) and cultures were performed on blood samples.ResultsThere were 59 (27.8%) cats seroreactive to one or more microorganisms. Considering cross-reactions, the seroprevalences were 15.6%-19.5% (R. massiliae-Bar29), 1.9%-6.2% (R. conorii), and 5.2%-7.5% (R. felis). A weak association was observed between SFG seropositivity and tick infestation. Ticks found on seropositive cats were Rhipicephalus pusillus, R. sanguineus and R. turanicus. DNA of Rickettsia was detected in 23 cats. 21 of them could be sequenced. Sequences obtained were identical to those sequences of SFG rickettsiae similar to R. conorii and R. massiliae. No amplification of R. felis was obtained.ConclusionsCats can be infected by SFG rickettsiae and produce antibodies against them. Cats may play a role in the transmission cycle of R. conorii and R. massiliae-Bar29, although the role in the R. felis cycle needs further analysis.