Tick-borne Spotted Fever Group Research Articles

A systematic review and meta-analysis of the prevalence of tick-borne SFGR in China from 2000 to 2022.

Ticks carry and transmit a wide range of pathogens (bacteria, viruses, and protozoa) that pose significant threats to human and animal health worldwide. Only few meta-analyses have been conducted on the distribution of ticks and tick-borne spotted fever group rickettsia (SFGR). Therefore, this study aims to examine the tick species and SFGR positivity in China in order to provide support for further research and improvements in the prevention and control of tick-borne diseases. This systematic review was performed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Relevant Chinese and English studies were retrieved from PubMed, Embase, Cochrane Library, Web of Science, China National Knowledge Infrastructure (CNKI), VIP database, Chinese Biomedical literature database (CBM) and Wanfang database from inception to January 9, 2023. Pooled SFGR positive rate was meta-analyzed using a random effects model and heterogeneity was assessed by the I2 index. Publication bias was also evaluated by funnel plot and Egger's test. The meta-analysis was performed on R studio 4.0.4. Meta-analysis of 57 studies published between 2000 and 2022 involving 39,380 ticks revealed a pooled SFGR positive rate of 21.4% (Q = 6423.74, I2 = 99%, Q-p<0.001, 95% CI: 15.0-29.6). Most studies of tick-borne SFGR infection rate were conducted in forest areas and developed animal husbandry areas in the northern region. There were slightly more tick species in the southern region, but the differences in tick species (Feeding tick 31.5%, 95%CI: 15.7-53.2, Questing tick 11.5%, 95%CI: 4.4-26.7, Q = 3.29, Q-p = 0.19) between areas (Northern area 20.4%, 95%CI: 14.1-28.7, Southern area 25.5%, 95%CI: 15.0-29.6, Q = 0.21, Q-p = 0.64) were not statistically significant. The most common tick species were Dermacentor silvarum (13%), Ixodes persulcatus (11%) and Haemaphysalis Iongicornis (10%), and the most prevalent SFGR species were Rickettsia raoultii (20%), Rickettsia heilongjiangiensis (11%), and some uncultured species (18%). This study examined the distribution of tick-borne SFGR in China. Our findings revealed that the main tick species were D. silvarum, I. persulcatus and H. iongicornis, and the common SFGR species were R. raoultii, R. heilongjiangiensis, and some uncultured species. Further studies are warranted to identify the potential vectors of SFGR and to better understand the epidemiology and pathogenesis of tick-borne diseases in China.

Review of Epidemiological Situation on Rickettsioses in the Russian Federation in 2022 as Compared with 2013–2021, Forecast for 2023

The aim of the review was to characterize the epidemiological situation on tick-borne rickettsioses (TR) in the Russian Federation in 2022 in comparison with the pandemic years of 2020–2021 and the pre-pandemic period of 2013–2019, to give the forecast for 2023. During 2013–2022, a pattern was observed in the incidence of rickettsiosis of the tick-borne spotted fever group (with the exception of Astrakhan spotted fever – ASF), that is also characteristic of most other natural focal infections (NFIs): a sharp decrease in registered incidence rates in 2020–2021 with the subsequent increase in 2022 almost up to pre-pandemic levels or even exceedance in some regions. The absence of a defining influence of the frequency of contacts of the population with vectors on the decrease in the incidence in the period of 2020–2021 and the rise in the value in 2022 confirms the registration-related nature of the changes. Given due attention to the problem of effective detection and reporting of NFIs, we expect the incidence of tick-borne rickettsioses to remain within the confidence intervals of long-term average values in most endemic regions in 2023. Relaxed vigilance to the problem of NFIs may lead to a decline in the reported incidence of Siberian tick-borne typhus in the Krasnoyarsk and Zabaikalsky Territories, Republics of Tuva and Khakassia, Novosibirsk and Amur Regions; reduction in the incidence of ASF in Astrakhan Region. The main issue in detecting and registration of the tick-borne rickettsiosis incidence, objective assessment and forecasting of the epidemic situation as regards this group of infections is conditioned by the lack of domestic certified diagnostic test kits for laboratory verification of rickettsioses. Close cooperation between epidemiologists and clinicians is required to establish and register cases of TR based on clinical and epidemiological data.

Tick-borne infections in the south of Russia: modern epidemiological situation, new approach to create “forecasting” and “explaining” morbidity models (in astrakhan rickettsiosis fever and crimean-congo hemorrhagic fever)

The article presents a description of the current tick-borne infection epidemiological situation in the south of Russia from the years 2013 to 2022, proposes a new approach to develop forecasting models for morbidity dynamics of Astrakhan rickettsial fever (ARF) and Crimean hemorrhagic fever (СCHF) in the Astrakhan region and presents data assessing 2022 explaining models for the Stavropol Territory and Astrakhan Region. Materials and methods. A comprehensive research was performed using epidemiological analysis and non-parametric statistical methods. The data assessing tick-borne infections epidemic process manifestations were retrieved from ARF and CCHF morbidity databases (developed as a project) and documents of infectious disease focus epidemiological examination provided by the departments of Rospotrebnadzor in the subjects of the Southern and North Caucasian Federal Districts. Morbidity models were developed using the Bayes theorem and Walds sequential statistical analysis, with a preliminary calculation of indicators informativeness by the Kullback method. The values of climatic factors from the database of the Center for Collective Use IKI-monitoring of the Space Research Institute of the Russian Academy of Sciences were used. Results. The results of the study indicate persistence of serious epidemiological situation regarding rickettsiosis of the tick-borne spotted fever group, Q fever, tick-borne borreliosis and CCHF in the south of Russia. Almost all tick-borne infections nosological forms in children under 14 years (including young children and infants) were widely involved in the epidemic process, which belong to patients at risk for a complicated disease course due to complicated diagnostics and treatment. The annual registration of tick-borne infections cases in the resort areas, with the subsequent occurrence of imported cases in other, including non-endemic regions poses a serious problem. The proposed forecasting models allow to predict the CСHF and ARF morbidity for each administrative district of the Astrakhan region with up to 91.7% accuracy. The explaining models CСHF accuracy for the Stavropol Territory and Astrakhan Region, when tested in 2022, was 88.5 and 83.3%, respectively, for ARF 91.7%. Conclusions. The further continuation of forecasting and explaining models verification for planning preventive measures and propose similar steps for tick-borne borreliosis and Q fever to epidemiological tick-borne infections to stabilize situation in the south of Russia.

Knowledge, Attitude and Practices among Pastoralists and Animal Health Workers on Tick-Borne Spotted Fever Group Rickettsiosis in Plateau State, Nigeria

Abstract Tick-borne Spotted Fever Group Rickettsioses (SFGR) is a serious disease reported in low income communities of many developed countries. The dearth of information on the existence of SFGR in most developing countries particularly in Sub Saharan Africa (SSA) including Nigeria, has been attributed to many factors chiefly amongst which are; poor knowledge of the disease, limitation of diagnostic capacity, leading to misdiagnosis and under reporting of some infectious zoonotic diseases such as SFGR. The aim of this study was to evaluate the pastoralists (livestock farmers) and animal health workers’ knowledge, attitudes and perceptions/practices towards ticks and tick-borne SFGR affecting livestock in 3 Local Government Areas (LGA): i.e., in Plateau State, Nigeria and including Jos the Capital City of the State. A total of 141 semi-structured questionnaires were administered to respondents including 90 pastoralists and 51 health workers from different veterinary clinics. Interestingly, the pastoralists expressed no knowledge about SFGR, although 61.36 % reported a history of tick infestation. Among the animal health workers, about 76.47 % expressed some knowledge about the disease, but lacked knowledge of the diagnosis of the disease. The findings indicate that there was the need for an increase awareness on SFGR and its predisposing factors among relevant stakeholders in the Plateau State, Nigeria.

Emerging tick-borne spotted fever group rickettsioses in the Balkans.

The impact of tick-borne pathogens (TBPs) on human health has increased in the last decades, since the incidence of emerging and re-emerging infectious and zoonotic tick-borne diseases has increased worldwide. Tick-borne rickettsiae of the Spotted Fever group (SFGR) are considered as emerging pathogens that can infect humans and cause a variety of non-specific clinical symptoms. Here, we report nine cases of atypical tick-borne diseases (9/460; 1.95%) that occurred over a period of four months (from 15 April 2021 to 16 August 2021) in Serbia, from which five cases were classified as confirmed SFGR infection, two cases as probable SFGR infection and two cases as suspected SFGR infection. Within cases of confirmed SFGR infection, R. helvetica was detected as the causative agent in two cases. The most common clinical finding was non-expanding persistent circular redness, followed by eschar and enlargement of regional lymph nodes, and pain at lesion site. Rickettsia outer membrane protein B (ompB) and citrate synthase (gltA) gene fragments were amplified from clinical samples and ticks attached to patients and IgG reacting with Rickettsia conorii antigen were detected in sera samples of patients, which are highly suggestive of exposure to SFGR. Surveillance and monitoring of rickettsial diseases in Serbia should continue and extended to new areas due to the increasing trend of clinical infections caused by SFGR in the country.

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.

Epidemiological Situation on Tick-Borne Spotted Fever Group Rickettsioses in the Russian Federation in 2012–2021, Prognosis for 2022–2026

The aim of the review was to characterize the epidemiological situation on rickettsiosis of the tick-borne spotted fever (TSF) group in the constituent entities of the Russian Federation in 2012–2021 and provide a long-term forecast for 2022–2026. In 2020–2021, indicators of registered incidence of tick-borne rickettsiosis in Russia on the whole decreased relative to the average long-term pre-pandemic level in 2012–2019 by 2 fold for all forms of the TSF group on average (Siberian tick-borne typhus – STT, Astrakhan spotted fever and Mediterranean fever). A comparative analysis of the territories revealed a correlation between the incidence of STT and the number of medical facility visits due to “tick bites” (r=0.67, p=0.015), which, against the background of an increase in the frequency of contacts of the population with vectors in 2020–2021 in most regions and a decrease in the incidence in the same regions indicates the credible nature of the changes. The ranking of territories according to average long-term incidence rates made it possible to classify the Republic of Altai and the Altai Territory as regions of extremely high epidemic hazard as regards STT; high hazard – the Republic of Tuva; above average – the Republic of Khakassia, Khabarovsk Territory and the Jewish Autonomous Region. The established downward trend in the incidence of tick-borne rickettsiosis among the population will be sustained in most entities of the Russian Federation within the next 5 years, excluding the Republic of Altai, the Altai Territory, the Republic of Crimea and the city of Sevastopol, where current incidence or an increase should be expected. The results of the analysis indicate the need to put more emphasis on natural-focal infections at the time of COVID-19 pandemic.

Seroepidemiological Study of Spotted Fever Group Rickettsiae and Identification of a Putative New Species, Rickesttsia sp. Da-1, in Gongliao, Northeast Taiwan.

Tick-borne spotted fever group (SFG) rickettsioses were neglected in Taiwan. The study reported a seroepidemiological survey of SFG rickettsiae in residents in Gongliao District, Northeast Taiwan. Blood samples were examined for antibodies against SFG rickettsiae by enzyme-linked immunosorbent assay and immunofluorescence assay. Risk factors were assessed using logistic regression. Ticks parasitizing dogs were collected within a 2 km radius from the houses of seropositive participants, and PCR was performed to detect possible tick-borne pathogens. Of 1108 participants, 75 (6.8%) had antibodies against SFG rickettsiae. Residents were more likely to be seropositive if they were older than 65 years, recruited by Dr. Enjoy’s Clinic, or resided in Jilin village. A total of 184 ticks including 5 species (Rhipicephalus sanguineus, Rhipicephalus haemaphysaloides, Dermacentor auratus, Haemaphysalis hystricis, Haemaphysalis ornithophila) were collected. Rickettsia spp. were detected in 6.5% (12/184) of ticks. Rickettsia sp. TwKM01 was found in 6 R. sanguineus and 4 R. haemaphysaloides; while Rickettsia sp. TwKM03 was identified in 1 R. sanguineus. Moreover, gene-based pairwise analysis indicated identification of a putative new species, Rickettsia sp. Da-1, in D. auratus. These findings provided evidence of SFG rickettsiae infection in ticks and suggested SFG rickettsiae exposure in the residents.

Amplicon-Based Next Generation Sequencing for Rapid Identification of Rickettsia and Ectoparasite Species from Entomological Surveillance in Thailand.

Background: Next generation sequencing (NGS) technology has been used for a wide range of epidemiological and surveillance studies. Here, we used amplicon-based NGS to species identify Rickettsia and their arthropod hosts from entomological surveillance. Methods: During 2015–2016, we screened 1825 samples of rodents and ectoparasites collected from rodents and domestic mammals (dog, cat, and cattle) across Thailand for Rickettsia. The citrate synthase gene was amplified to identify Rickettsia to species, while the Cytochrome Oxidase subunit I (COI) and subunit II (COII) genes were used as target genes for ectoparasite identification. All target gene amplicons were pooled for library preparation and sequenced with Illumina MiSeq platform. Result: The highest percentage of Rickettsia DNA was observed in fleas collected from domestic animals (56%) predominantly dogs. Only a few samples of ticks from domestic animals, rodent fleas, and rodent tissue were positive for Rickettisia DNA. NGS based characterization of Rickettsia by host identified Rickettsia asembonensis as the most common bacteria in positive fleas collected from dogs (83.2%) while “Candidatus Rickettsia senegalensis” was detected in only 16.8% of Rickettsia positive dog fleas. Sequence analysis of COI and COII revealed that almost all fleas collected from dogs were Ctenocephalides felis orientis. Other Rickettsia species were detected by NGS including Rickettsia heilongjiangensis from two Haemaphysalis hystricis ticks, and Rickettsia typhi in two rodent tissue samples. Conclusion: This study demonstrates the utility of NGS for high-throughput sequencing in the species characterization/identification of bacteria and ectoparasite for entomological surveillance of rickettsiae. A high percentage of C. f. orientis are positive for R. asembonensis. In addition, our findings indicate there is a risk of tick-borne Spotted Fever Group rickettsiosis, and flea-borne murine typhus transmission in Tak and Phangnga provinces of Thailand.

Ticks and rickettsial exposure in lowland tapirs (Tapirus terrestris) of three Brazilian biomes

This study evaluated ticks and rickettsial exposure in 220 free-ranging lowland tapirs, Tapirus terrestris, from 2006 to 2018 in selected areas of three major biomes of Brazil – Atlantic Forest, Pantanal, and Cerrado. Overall, a total of 5970 tick specimens representing the following nine species were collected from tapirs: Amblyomma brasiliense, Amblyomma coelebs, Amblyomma dubitatum, Amblyomma ovale, Amblyomma parvum, Amblyomma sculptum, Amblyomma triste, Haemaphysalis juxtakochi, and Rhipicephalus microplus. Amblyomma sculptum was the most prevalent and abundant tick species in all three biomes; however, mean intensity values for A. sculptum were significantly lower in Atlantic Forest than in the Pantanal or Cerrado, and at the same time, statistically similar among tapirs from Pantanal and Cerrado. Contrastingly, mean intensity values for A. coelebs were significantly higher in the Atlantic Forest than in the other biomes. The remaining tick species were collected in lower numbers, or were exclusive for one biome, e.g., A. brasiliense and H. juxtakochi only in the Atlantic Forest. A total of 177 blood sera (123 individuals plus 54 recaptures) were collected from tapirs and tested for the presence of reactive antibodies to six Rickettsia species by immunofluorescence assay. Overall, 69% (9/13), 49% (62/126), and 66% (25/38) tapir sera from Atlantic Forest, Pantanal and Cerrado, respectively, were seroreactive to at least one Rickettsia species, with no significant difference between the three areas. Although many tapir sera reacted simultaneously to ≥2 Rickettsia species, Rickettsia parkeri elicited highest % seroprevalence and endpoint titers, and was incriminated as the possible agent involved in a homologous reaction in tapirs from the three biomes, where A. ovale was previously found infected by R. parkeri. In fact, seroconversion to R. parkeri was demonstrated in five tapirs that were captured at least twice during the study. This study demonstrated that tapirs were found to be constantly infested by several tick species in the Atlantic Forest, Pantanal and Cerrado biomes; however, the richness of tick infestations was concordant to the tick species known to be established in each biome. Under natural conditions, lowland tapirs were shown to be exposed to tick-borne spotted fever group rickettsiae.

Identification and distribution of nine tick-borne spotted fever group Rickettsiae in the Country of Georgia

Rickettsial pathogens cause diseases that vary in severity and clinical presentation. Rickettsia species transmitted by ticks are mostly classified within the spotted fever group of rickettsiae (SFGR) and are often associated with febrile diseases. Preliminary studies have detected three human-pathogenic SFGR from ticks in Georgia: Rickettsia aeschlimannii, Rickettsia raoultii, and Rickettsia slovaca. To more broadly assess the presence of tick-borne rickettsiae from Georgia we examined 1594 ticks, representing 18 species from five genera (Ixodes, Hyalomma, Haemaphysalis, Dermacentor, and Rhipicephalus), collected from eight regions of Georgia. A total of 498 tick DNA samples extracted from single ticks or pooled ticks were assessed by molecular methods. Genus-specific Rick17b and species-specific qPCR assays were used to identify six rickettsiae: R. aeschlimannii, R. raoultii, R. slovaca, Rickettsia conorii subsp. conorii, Rickettsia massiliae, and Rickettsia monacensis. Tick samples that were positive for Rickettsia, but not identified by the species-specific assays, were further evaluated by multi-locus sequence typing (MLST) using sequences of four protein-coding genes (gltA, ompA,ompB, sca4). Three additional Rickettsia species were identified by MLST: Candidatus Rickettsia barbariae, Rickettsia helvetica, and Rickettsia hoogstraalii. Overall, nine species of Rickettsia (six human pathogens and three species with unknown pathogenicity) were detected from 12 tick species of five different genera. A distribution map for the tick-borne rickettsiae revealed six newly identified endemic regions in Georgia.

Diversity of Rickettsia in Ticks Collected in Abruzzi and Molise Regions (Central Italy).

Rickettsiae have worldwide occurrence and rickettsiosis are widely recognized as emerging infections in several parts of the world. For decades, it was thought that a single pathogenic tick-borne spotted fever group (SFG), Rickettsia, occurred in each continent. Nowadays, thanks to molecular biology, new species of Rickettsia responsible for disease in humans are continuously identified worldwide. In a framework of diagnostic activities of the Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Gaporale” and considering some reports of suspected human clinical cases of rickettsiosis, a survey on ticks collected form animals and humans was carried out with the aim to identify the Rickettsia species circulating in Abruzzi and Molise regions. A total of 603 ticks, previously identified at species level by morphology, pooled into 178 tick samples, were tested by pan-Rickettsia RealTime PCR. DNA from specimens positive for Rickettsia spp. was then sequenced in order to identify the Rickettsia species involved. The highest infection rate was detected in Dermacentor marginatus followed by Ixodes ricinus. The selected targets for this purpose were OmpA and gltA. Rickettsia slovaca, Rickettsia monacensis, Rickettsia massiliae, Rickettsia conorii, Rickettsia aeschlimannii, Rickettsia helvetica, Rickettsia raoultii, and Rickettsia felis – like organisms were identified in this study. These are the first data available in the literature for the circulation of SFG Rickettsia species in the selected geographical area. Results made evidence of high rate of infection in ticks. All Rickettsia species detected have been previously involved in human infection. The diversity of Rickettsia detected, and tick species collected reflects the biodiversity in term of wildlife and environment of the area. An association between Rickettsia species, ticks, and the relationships with vertebrate host species are discussed. Due to the peculiar eco-biology of each Rickettsia species, the use of diagnostic tools able to identify Rickettsia at the species level is thus recommended in order to assess the risk for humans and to elucidate more precise etiological diagnosis in clinical cases.

Molecular detection of spotted fever group rickettsiae in ticks from Cameroon

In western and eastern Africa, rickettsioses are one cause of fever in humans. Little is known regarding the presence of Rickettsia sp. in northern Cameroon. The present work was conducted in order to identify potential tick-borne spotted fever group Rickettsia in the Adamawa region of northern Cameroon, which may contribute filling some of the knowledge gaps of these pathogens.Ticks were collected from cattle in the municipal slaughterhouse of Ngaoundere in the Adamawa region of northern Cameroon. After morphological identification of tick species, extracted DNA was analyzed by PCR targeting the rickettsial ompB gene and the intergenic spacers dksA-xerC, mppA-purC and rpmE-tRNAfMet.Of the 316 adult ticks collected, 149 (47.1%) were Amblyomma variegatum, 92 (29%) Rhipicephalus spp. and 75 (23.7%) Hyalomma spp. Through the use of conventional PCR assays for the rickettsial ompB gene, rickettsial DNA was detected in 104 (32.9%) samples (85 Amblyomma sp., 14 Hyalomma spp. and 5 Rhipicephalus spp.). The ompB gene and the three intergenic were sequenced for 10 ticks in order to determine the rickettsial species. Rickettsia africae was detected in Amblyomma variegatum, Rickettsia aeschlimannii in Hyalomma rufipes and Hyalomma truncatum, Rickettsia sibirica in H. truncatum, Rickettsia massiliae in Rhipicephalus lunulatus and Candidatus Rickettsia barbariae in R. lunulatus. To the best of the author’s knowledge, this report represents the first molecular evidence of rickettsial infection in ticks in the Adamawa region of northern Cameroon, which suggests a possible exposure of the human population in this region.

Molecular evidence of Rickettsia spp. in ixodid ticks and rodents in suburban, natural and rural habitats in Slovakia

BackgroundNatural foci of tick-borne spotted fever group (SFG) rickettsiae of public health concern have been found in Slovakia, but the role of rodents in their circulation is unclear. Ticks (Ixodes ricinus, Ixodes trianguliceps, Dermacentor marginatus, Dermacentor reticulatus, Haemaphysalis concinna and Haemaphysalis inermis) and tissues of rodents (Apodemus flavicollis, Apodemus sylvaticus, Myodes glareolus, Microtus arvalis, Microtus subterraneus and Micromys minutus) were examined for the presence of SFG rickettsiae and Coxiella burnetii by molecular methods. Suburban, natural and rural habitats were monitored to acquire information on the role of ticks and rodents in the agents’ maintenance in various habitat types of Slovakia.ResultsThe overall prevalence of rickettsial infection in questing I. ricinus and D. marginatus was 6.6% and 21.4%, respectively. Rickettsia helvetica, R. monacensis and non-identified rickettsial species were detected in I. ricinus, whereas R. slovaca and R. raoultii were identified in D. marginatus. Rickettsia spp.-infected I. ricinus occurred during the whole tick questing period. Rickettsia helvetica dominated (80.5%) followed by R. monacensis (6.5%). The species were present in all studied habitats. Rickettsia slovaca (66.7%) and R. raoultii (33.3%) were identified in D. marginatus from the rural habitat. Apodemus flavicollis was the most infested rodent species with I. ricinus, but My. glareolus carried the highest proportion of Rickettsia-positive I. ricinus larvae. Only 0.5% of rodents (A. flavicollis) and 5.2% of engorged I. ricinus removed from My. glareolus, A. flavicollis and M. arvalis were R. helvetica- and R. monacensis-positive. Coxiella burnetii was not detected in any of the tested samples. We hypothesize that rodents could play a role as carriers of infected ticks and contribute to the maintenance of rickettsial pathogens in natural foci.ConclusionsLong-term presence of SFG Rickettsia spp. was confirmed in questing ticks from different habitat types of Slovakia. The results suggest a human risk for infection with the pathogenic R. helvetica, R. monacensis, R. slovaca and R. raoultii.

Comparative value of blood and skin samples for diagnosis of spotted fever group rickettsial infection in model animals

The definitive diagnosis of spotted fever group (SFG) rickettsioses in humans is challenging due to the retrospective nature and cross reactivity of the serological methods and the absence of reliable and consistent samples for molecular diagnostics. Existing data indicate the transient character of bacteremia in experimentally infected animals. The ability of arthropod vectors to acquire rickettsial infection from the laboratory animals in the absence of systemic infection and known tropism of rickettsial agents to endothelial cells of peripheral blood vessels underline the importance of local infection and consequently the diagnostic potential of skin samples. In order to evaluate the diagnostic sensitivity of rickettsial DNA detection in blood and skin samples, we compared results of PCR testing in parallel samples collected from model laboratory animals infected with Rickettsia rickettsii, Rickettsia parkeri and Rickettsia slovaca-like agent at different time points after infection. Skin samples were collected from ears – away from the site of tick placement and without eschars. Overall, testing of skin samples resulted in a higher proportion of positive results than testing of blood samples. Presented data from model animals demonstrates that testing of skin samples from sites of rickettsial proliferation can provide definitive molecular diagnosis of up to 60–70% of tick-borne SFG rickettsial infections during the acute stage of illness. Detection of pathogen DNA in cutaneous samples is a valuable alternative to blood-PCR at least in model animals.

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.

Rickettsioses as causes of CNS infection in southeast Asia.

1report that scrub typhus caused by Orientia tsutsugamushi, murine typhus caused by Rickettsia typhi, and leptospirosis caused by various Leptospira species account for more than a third of CNS infections diagnosed over 8 years in Vientiane Hospital in Laos. The study is one more great contribution from this team in their investigation of undocumented syndromes, as well as in the public health challenge of rickettsial diseases in southeast Asia. The same investigators have previously reported that scrub typhus was the second most common microbial cause of fever of unknown origin in rural Laos (122 [15%] of 799 diagnosed cases). 2 In 2006, rickettsial infection was detected in 115 (27%) of 427 adults admitted to Vientiane Hospital for fever with negative blood culture. 3 The most common rickettsial agent was O tsutsugamushi followed by R typhi. Fewer data are available about the prevalence of these diseases in other southeast Asian countries. In Thailand, scrub and murine typhus has been reported 4 in 16% and 2%, respectively, of fever of unknown origin, with mortality of 3–17% for scrub typhus. Even if epidemiological data for the whole region are unavailable, the substantial presence of rickettsial infections is shown by frequent reports in travellers returning from this area. 5

High prevalence of Rickettsia africae variants in Amblyomma variegatum ticks from domestic mammals in rural western Kenya: implications for human health.

Tick-borne spotted fever group (SFG) rickettsioses are emerging human diseases caused by obligate intracellular Gram-negative bacteria of the genus Rickettsia. Despite being important causes of systemic febrile illnesses in travelers returning from sub-Saharan Africa, little is known about the reservoir hosts of these pathogens. We conducted surveys for rickettsiae in domestic animals and ticks in a rural setting in western Kenya. Of the 100 serum specimens tested from each species of domestic ruminant 43% of goats, 23% of sheep, and 1% of cattle had immunoglobulin G (IgG) antibodies to the SFG rickettsiae. None of these sera were positive for IgG against typhus group rickettsiae. We detected Rickettsia africae-genotype DNA in 92.6% of adult Amblyomma variegatum ticks collected from domestic ruminants, but found no evidence of the pathogen in blood specimens from cattle, goats, or sheep. Sequencing of a subset of 21 rickettsia-positive ticks revealed R. africae variants in 95.2% (20/21) of ticks tested. Our findings show a high prevalence of R. africae variants in A. variegatum ticks in western Kenya, which may represent a low disease risk for humans. This may provide a possible explanation for the lack of African tick-bite fever cases among febrile patients in Kenya.

Novel identification of Dermacentor variabilis Arp2/3 complex and its role in rickettsial infection of the arthropod vector.

Tick-borne spotted fever group (SFG) Rickettsia species must be able to infect both vertebrate and arthropod host cells. The host actin-related protein 2/3 (Arp2/3) complex is important in the invasion process and actin-based motility for several intracellular bacteria, including SFG Rickettsia in Drosophila and mammalian cells. To investigate the role of the tick Arp2/3 complex in tick-Rickettsia interactions, open reading frames of all subunits of the protein including Arp2, Arp3, ARPC1, ARPC2, ARPC3, ARPC4, and ARPC5 were identified from Dermacentor variabilis. Amino acid sequence analysis showed variation (ranging from 25–88%) in percent identity compared to the corresponding subunits of the complex from Drosophila melanogaster, Mus musculus, Homo sapiens, and Saccharomyces cerevisiae. Potential ATP binding sites were identified in D. variabilis (Dv) Arp2 and Arp3 subunits as well as five putative WD (Trp-Asp) motifs which were observed in DvARPC1. Transcriptional profiles of all subunits of the DvArp2/3 complex revealed greater mRNA expression in both Rickettsia-infected and -uninfected ovary compared to midgut and salivary glands. In response to R. montanensis infection of the tick ovary, the mRNA level of only DvARPC4 was significantly upregulated compared to uninfected tissues. Arp2/3 complex inhibition bioassays resulted in a decrease in the ability of R. montanensis to invade tick tissues with a significant difference in the tick ovary, indicating a role for the Arp2/3 complex in rickettsial invasion of tick cells. Characterization of tick-derived molecules associated with rickettsial infection is imperative in order to better comprehend the ecology of tick-borne rickettsial diseases.

Importation of exotic ticks and tick-borne spotted fever group rickettsiae into the United States by migrating songbirds

Birds are capable of carrying ticks and, consequently, tick-transmitted microorganisms over long distances and across geographical barriers such as oceans and deserts. Ticks are hosts for several species of spotted fever group rickettsiae (SFGR), which can be transmitted to vertebrates during blood meals. In this study, the prevalence of this group of rickettsiae was examined in ticks infesting migratory songbirds by using polymerase chain reaction (PCR). During the 2009 and 2010 spring migration season, 2064 northward-migrating passerine songbirds were examined for ticks at Johnson Bayou, Louisiana. A total of 91 ticks was removed from 35 individual songbirds for tick species identification and spotted fever group rickettsia detection. Ticks were identified as Haemaphysalis juxtakochi (n=38, 42%), Amblyomma longirostre (n=22, 24%), Amblyomma nodosum (n=17, 19%), Amblyomma calcaratum (n=11, 12%), Amblyomma maculatum (n=2, 2%), and Haemaphysalis leporispalustris (n=1, 1%) by comparing their 12S rDNA gene sequence to homologous sequences in GenBank. Most of the identified ticks were exotic species originating outside of the United States. The phylogenetic analysis of the 71 ompA gene sequences of the rickettsial strains detected in the ticks revealed the occurrence of 6 distinct rickettsial genotypes. Two genotypes (corresponding to a total of 28 samples) were included in the Candidatus Rickettsia amblyommii clade (less than 1% divergence), 2 of them (corresponding to a total of 14 samples) clustered with Rickettsia sp. “Argentina” with less than 0.2% sequence divergence, and 2 of them (corresponding to a total of 27 samples), although closely related to the R. parkeri–R. africae lineage (2.50–3.41% divergence), exhibited sufficient genetic divergence from its members to possibly constitute a new rickettsial genotype. Overall, there does not seem to be a specific relationship between exotic tick species, the rickettsiae they harbor, or the reservoir competence of the corresponding bird species.