Differential interactions of Rickettsia species with tick microbiota in Rh. sanguineus and Rh. turanicus.

Tick-borne rickettsioses, caused by Gram-negative bacteria of the Rickettsia genus, pose a growing global threat, with various arthropod vectors contributing to their transmission. Understanding the complex interactions within tick microbiota, including the role of Rickettsia species, is crucial for elucidating the dynamics of rickettsial diseases. Here, we investigate the taxonomic profiles and co-occurrence networks of Rickettsia in Rh. sanguineus sensus lato (s.l.) and Rh. turanicus ticks, revealing significant differences in community composition and local connectivity of Rickettsia species. While the microbiota of both tick species share common taxa, distinct differences in relative abundance and network topology suggest unique ecological niches. Moreover, robustness analysis demonstrates varying resilience to perturbations, indicating different strategies for network organization. Our findings also highlight metabolic differences between tick species, suggesting potential implications for Rickettsia interactions. Overall, this study provides insights into the intricate microbial landscape within ticks, shedding light on the functional redundancy and metabolic pathways associated with Rickettsia, thus advancing our understanding of tick-borne diseases.

[First autochthonous case of spotted fever associated with the Reserva Ecológica Costanera Sur, Buenos Aires City]. / Primer caso autóctono de fiebre manchada asociado a la Reserva Ecológica Costanera Sur, Ciudad Autónoma de Buenos Aires.

The first autochthonous case of rickettsiosis is reported here. The case occurred in the Costanera Sur Ecological Reserve, a protected area of the City of Buenos Aires, in August 2022, where 4 species of ticks were found, namely Amblyomma aureolatum, Ixodes auritulus sensu lato, Rhipicephalus sanguineus sensu stricto and Amblyomma triste. The epidemiological, ecological, clinical and laboratory aspects that allowed timely diagnosis and appropriate treatment are also described.

Island hitchhikers: pathogen agents of Madeira and Azores ticks.

Ticks are blood-sucking arthropods that can transmit pathogens to their host. As insular ecosystems can enhance tick-host interactions, this study aimed to understand tick diversity, pathogen presence, and their respective associations in the Azores and Madeira archipelagos. Unfed or partially engorged ticks (n = 120) were collected from 58 cats and dogs in the Azores (n = 41 specimens) and Madeira (n = 79 specimens) from November 2018 to March 2019. Vector identification was based on morphology and molecular criteria. For pathogen sequencing, 18S gene fragment for Babesia/Hepatozoon and gltA for Rickettsia were performed. Sequence data was explored using BLAST and BLAST and phylogenetic inference tools. In the Azores, Ixodes hexagonus, I. ventalloi, and Rhipicephalus sanguineus (n = 6; 14.6%, n = 6; 14.6%, and n = 29; 70.7% respectively) were found and in Madeira I. ricinus and R. sanguineus (n = 78, 98.7%; and n = 1, 1.3%; respectively) were identified. Tick COI markers confirmed species highlighting confirmation of R. sanguineus s.s. and genotype A of I. ventalloi. In the Azores Islands, the detected Rickettsia massiliae was linked to R. sanguineus (dogs and cats) and I. hexagonus (dogs), and in Madeira Island, R. monacensis (dogs) and Hepatozoon silvestris (cats) were found associated with I. ricinus. Further, I. ventalloi presence in the Azores expands west its known range, and Hepatozoon silvestris in Madeira may suggest that I. ricinus could have a role as a potential vector. Finally, as R. massiliae and R. monacensis presence underlines public health risks, surveillance by health authorities is crucial as pathogen-tick interactions may drive disease spread, therefore monitoring remains pivotal for disease prevention.

Endosymbiont-derived metabolites are essential for tick host reproductive fitness.

Ticks, like other obligatory blood-feeding arthropods, rely on endosymbiotic bacteria to supplement their diet with B vitamins lacking in blood. It has been suggested that additional metabolites such as L-proline may be involved in this nutritional symbiosis, but this has yet to be tested. Here, we studied the metabolite-based interaction between the brown dog tick Rhipicephalus sanguineus (Acari: Ixodidae) and its Coxiella-like endosymbionts (CLE). We measured amino acid titers and tested the effect of B vitamins and L-proline supplementation on the fitness of CLE-suppressed female ticks, displaying low titers of CLE. We found higher titers of L-proline in the symbiont-hosting organs of unfed ticks and in engorged blood-fed whole ticks. Supplementation of B vitamins increased the hatching rate of CLE-suppressed ticks; this effect appears to be stronger when L-proline is added. Our results indicate that L-proline is produced by CLE, and we suggest that CLE is essential in states of high metabolic demand that affects tick reproductive fitness, such as oogenesis and embryonic development. These findings demonstrate the broader effect of nutritional symbionts on their hosts and may potentially contribute to the control of ticks and tick-borne diseases. IMPORTANCE: Coxiella-like endosymbionts (CLE) are essential to the brown dog tick Rhipicephalus sanguineus for feeding and reproduction. This symbiosis is based on the supplementation of B vitamins lacking in the blood diet. The involvement of additional metabolites has been suggested, but no experimental evidence is available as yet to confirm a metabolic interaction. Here, we show that B vitamins and L-proline, both of which contribute to tick reproductive fitness, are produced by CLE. These findings demonstrate the importance of symbiont-derived metabolites for the host's persistence and shed light on the complex bacteria-host metabolic interaction, which can be channeled to manipulate and control tick populations.

The case of Mediterranean spotted fever of the traveler returned from Zambia.

We report the case of a traveler who returned from Zambia and was diagnosed with Mediterranean spotted fever (MSF), an infectious disease caused by Rickettsia conorii conorii. The patient presented to Sapporo City General Hospital with symptoms of fever, malaise, headache, and rash. The pathogen was identified by Polymerase Chain Reaction assays and subsequent analyses. The patient improved with 10-day treatment of oral doxycycline. Although some cases of MSF have been reported in sub-Saharan Africa, none have been reported in Zambia. Rhipicephalus sanguineus sensu lato, the vector of the Rickettsia conorii conorii, has been found in various areas of Zambia. Our case report highlights the potential threat of Mediterranean spotted fever in urban areas of Zambia.

Rhipicephalus sanguineus from Hungarian dogs: Tick identification and detection of tick-borne pathogens.

The brown dog tick, Rhipicephalus sanguineus is a complex of tick species with an unsettled species concept. In Europe, R. sanguineus is considered mainly a Mediterranean tick with sporadic findings in central and northern Europe. R. sanguineus is known as a vector of a range of pathogens of medical and veterinary importance, most of which not yet reported as autochthonous in Hungary. A total of 1839 ticks collected by veterinarians from dogs and cats were obtained in Hungary. The study aims at precise determination of ticks identified as R. sanguineus and detection of pathogens in collected ticks. All ticks were morphologically determined and 169 individuals were identified as R. sanguineus. A subset of 15 ticks was selected for molecular analysis (16S rDNA, 12S rDNA, COI). Phylogenetic analyses invariably placed sequences of all three markers into a single haplotype identified as R. sanguineus sensu stricto. All 169 brown dog ticks were tested for the presence of A. platys, E. canis, R. conorii, B. vogeli and H. canis. None of the investigated ticks was positive for the screened pathogens, though A. phagocytophilum sequence was detected in a single tick.

Roaming Dogs, Intense Brown Dog Tick Infestation, and Emerging Rocky Mountain Spotted Fever in Tijuana, México.

A two decades-long epidemic of Rocky Mountain spotted fever in northern México reached the U.S. border city of Tijuana in 2021. Cases were near the city periphery in marginalized areas, some lacking infrastructure such as streets or utilities. We worked in the three census areas where human cases were reported and in 12 additional control Áreas Geoestadisticas Básicas. There were dogs, the primary tick host and Rickettsia rickettsii reservoir, in 76% of homes, with 2.2 owned dogs per home on average, approximately equal numbers of roaming dogs were seen, and 46.2% of owned dogs were allowed to roam in the street. Sixty-eight percent of people had heard of Rocky Mountain spotted fever (RMSF), and 35% self-reported tick infestation, including 19% of homes without dogs. Ticks appeared to move among houses of adjacent neighbors. Of 191 examined dogs, 61.8% were tick-infested, with 6-fold increased odds if they were allowed to roam. Although no dogs were Rickettsia polymerase chain reaction-positive, we found one R. rickettsii- and 11 Rickettsia massiliae-infected ticks. The rickettsial IgG seroprevalence by immunofluorescence antibody assay was 76.4%, associated with unhealthy body condition, adults, dogs with >10 ticks, more dogs being seen in the area, and dogs being permitted in the street. Insufficient medical and canine management resources have contributed to a case fatality rate of RMSF that has exceeded 50% in areas. High canine seroprevalence suggests risks to people and dogs; unfortunately, herd immunity is impeded by high turnover in the canine population owing to the birth of puppies and high death rates. Binational One Health workers should monitor disease spread, enact canine population management and tick eradication, and provide prevention, diagnostic, and treatment support.

Microfluidic PCR and network analysis reveals complex tick-borne pathogen interactions in the tropics.

BACKGROUND: Ixodid ticks, particularly Rhipicephalus sanguineus s.l., are important vectors of various disease-causing agents in dogs and humans in Cuba. However, our understading of interactions among tick-borne pathogens (TBPs) in infected dogs or the vector R. sanguineus s.l. remains limited. This study integrates microfluidic-based high-throughput real-time PCR data, Yule's Q statistic, and network analysis to elucidate pathogen-pathogen interactions in dogs and ticks in tropical western Cuba. METHODS: A cross-sectional study involving 46 client-owned dogs was conducted. Blood samples were collected from these dogs, and ticks infesting the same dogs were morphologically and molecularly identified. Nucleic acids were extracted from both canine blood and tick samples. Microfluidic-based high-throughput real-time PCR was employed to detect 25 bacterial species, 10 parasite species, 6 bacterial genera, and 4 parasite taxa, as well as to confirm the identity of the collected ticks. Validation was performed through end-point PCR assays and DNA sequencing analysis. Yule's Q statistic and network analysis were used to analyse the associations between different TBP species based on binary presence-absence data. RESULTS: The study revealed a high prevalence of TBPs in both dogs and R. sanguineus s.l., the only tick species found on the dogs. Hepatozoon canis and Ehrlichia canis were among the most common pathogens detected. Co-infections were observed, notably between E. canis and H. canis. Significant correlations were found between the presence of Anaplasma platys and H. canis in both dogs and ticks. A complex co-occurrence network among haemoparasite species was identified, highlighting potential facilitative and inhibitory roles. Notably, H. canis was found as a highly interconnected node, exhibiting significant positive associations with various taxa, including A. platys, and E. canis, suggesting facilitative interactions among these pathogens. Phylogenetic analysis showed genetic diversity in the detected TBPs. CONCLUSIONS: Overall, this research enhances our understanding of TBPs in Cuba, providing insights into their prevalence, associations, and genetic diversity, with implications for disease surveillance and management.

Molecular survey of canine tick-borne pathogens in ticks and stray dogs in Dhaka city, Bangladesh.

Molecular surveillance of canine tick-borne pathogens (TBPs) in Bangladesh has constantly been undervalued. Therefore, the emergence of new pathogens often remains undetected. This study aimed to screen tick-borne pathogens in stray dogs and ticks in the Dhaka metropolitan area (DMA). Eighty-five dog blood and 53 ticks were collected in six city districts of DMA from September 2022 to January 2023. The ticks were identified by morphology. Screening of TBPs was performed by polymerase chain reaction (PCR), followed by sequencing. The PCR assays were conducted to analyze the 18S rRNA (Babesia gibsoni, B. vogeli, and Hepatozoon canis), 16S rRNA (Anaplasma phagocytophilum, A. platys, and A. bovis), gltA (Ehrlichia canis and Rickettsia spp.), flagellin B (Borrelia spp.) and 16-23S rRNA (Bartonella spp.). Three tick species, Rhipicephalus sanguineus (50/53), R. microplus (1/53), and Haemaphysalis bispinosa (2/53), were identified. Babesia gibsoni (38 out of 85) and A. platys (7 out of 85) were detected in dog blood. In contrast, four pathogens, B. gibsoni (1 out of 53), B. vogeli (1 out of 53), H. canis (22 out of 53), and A. platys (1 out of 53), were detected in the ticks. However, the detection rates of TBPs in dog blood and ticks were not correlated in this study. The phylogenetic analyses suggested that a single genotype for each of the four pathogens is circulating in DMA. This study reports the existence of B. vogeli, H. canis, and A. platys in Bangladesh for the first time.

New records of Rhipicephalus linnaei infected by Rickettsia massiliae from Central Mexico.

BACKGROUND: On the American continent, Rhipicephalus sanguineus s.l. comprises two species: Rhipicephalus linnaei and R. sanguineus s.s. Each species has been identified as a potential vector of at least one of five species of pathogenic bacteria of the genus Rickettsia. In particular, Rickettsia massiliae is one of three species with the greatest importance in public health at the continental level. In Mexico, this species is reported exclusively in the Nearctic states of Baja California and Chihuahua. AIM: For this reason, the aim of this work was to provide new records of R. massiliae for the centre of the country derived from active acarological surveillance. MATERIALS AND METHODS: During the period of February-October 2019, 29 dogs from six municipalities in the state of Morelos were sampled. Hosts were visually inspected, and ticks were recovered and identified morphologically and molecularly by amplification of the 16S rDNA gene. Subsequently, five genes from members of the genus Rickettsia were amplified and sequenced. RESULTS: A total of 229 (117♀, 98♂ and 14 N) ticks identified as R. linnaei were recovered, two of which were positive for R. massiliae strains related to those recovered from Argentina and the United States. CONCLUSIONS: This work provides the second record of R. massiliae infecting R. linnaei in Mexico and the Americas, increasing the geographic distribution of this Rickettsia species in the Neotropical region, and providing information on the possible role of R. linnaei as a potential vector of this microorganism.

Optimal control of a tick population with a view to control of Rocky Mountain Spotted Fever.

In some regions of the Americas, domestic dogs are the host for the tick vector Rhipicephalus sanguineus, and spread the tick-borne pathogen Rickettsia rickettsii, which causes Rocky Mountain Spotted Fever (RMSF) in humans. Interventions are carried out against the vector via dog collars and acaricidal wall treatments. This paper investigates the optimal control of acaricidal wall treatments, using a prior model for populations and disease transmission developed for this particular vector, host, and pathogen. It is modified with a death term during questing stages reflecting the cost of control and level of coverage. In the presence of the control, the percentage of dogs and ticks infected with Ri. rickettsii decreases in a short period and remains suppressed for a longer period, including after treatment is discontinued. Risk of RMSF infection declines by 90% during this time. In the absence of re-application, infected tick and dog populations rebound, indicating the eventual need for repeated treatment.

POTENTIAL SHARED DISEASE RISK AMONG DOGS AND COYOTES (CANIS LATRANS) EXEMPLIFIED BY THE ECOLOGY OF RICKETTSIOSIS IN A ROCKY MOUNTAIN SPOTTED FEVER-EPIDEMIC REGION IN NORTHERN MEXICO.

Rocky Mountain spotted fever (RMSF), caused by the bacterium Rickettsia rickettsii, is a re-emerging tick-borne zoonosis in North America, with hundreds of human fatalities in multiple outbreaks in northern Mexico and the southwestern US in the past few decades. Free-roaming dogs are key because they are reservoirs for the pathogen and the main hosts of the brown dog tick (Rhipicephalus sanguineus), which vectors RMSF in this region. Because coyotes (Canis latrans) can be infected with R. rickettsii and infested with Rh. sanguineus, we hypothesized that space sharing among dogs and coyotes could enhance disease risks. In summer 2021, we captured and sampled 11 coyotes at two sites in Baja California, Mexico, near population centers with human cases of RMSF, and fitted seven individuals with GPS logging collars. We also tested tissue samples, sera, and ectoparasites for DNA of R. rickettsii with PCR and used serology to detect antibodies to R. rickettsii. Finally, we deployed an array of cameras to document dog-coyote interactions. Mean home range size was 40.37 km2. Both GPS and camera data showed considerable home range overlap both between individual coyotes and between coyotes and dogs. Coyotes were active in areas where dogs occur including the domestic interface surrounding human settlements. Although none of our samples were positive for R. rickettsii on PCR, 72.7% (8/11) of the samples were seropositive with titers ≥64. Our data confirm shared space use and risk of shared parasites and disease between coyotes and dogs.

First detection and genetic identification of Rickettsia spp. from ticks collected from rodents in north India.

BACKGROUND: Changing climatic conditions and invasion of ticks in urban areas have led to a greater number of cases of tick-borne diseases, thus, becoming a matter of increasing concern. Tick borne rickettsioses are one of the important emerging diseases worldwide. Knowledge of epidemiology of the vector and pathogen in the community is essential in order to understand and prevent the transmission of the disease to healthy population. METHODS: In our present study, we trapped rodents in selected areas of Chandigarh and Punjab in north India. The rodents were screened for the presence of ticks which were further screened for the presence of rickettsial agents. PCRs targeting 17 â€‹kDa and gltA genes were carried out followed by Sanger sequencing of the positive amplicons followed by phylogenetic analysis of the sequences. RESULTS: A total of 17 ticks were collected out of which one (Rhipicephalus sanguineus) was found to be harboring a Rickettsia sp. PCR targeting gltA and 17 â€‹kDa genes of rickettsia were put up and Sanger sequencing was performed. Phylogenetic analysis revealed the sequences to be closely related to Rickettsia rhipicephali. CONCLUSION: The current study establishes the presence of rickettsial agents in the community. Although Rickettsia rhipicephali is a non-pathogenic agent, the study encourages more vigorous community surveillance should be carried out in order to determine the exact burden of rickettsial agents in our community. To our knowledge, this is the first study reporting Rickettsia rhipicephali in India.

Tick-associated diseases identified from hunting dogs during the COVID-19 pandemic in a Mayan community in Yucatan, Mexico.

Background: Hunting activity in the Mayan communities has increased due to COVID-19 and domestic dogs have gained more importance. Due to their proximity to humans, domestic dogs are a bridge between tick-borne diseases (TBDs) and humans and their peri-domestic environment. In Mexico, and especially in rural regions, there were not adequate records of TBDs during the SARS-CoV-2 pandemic. Aim: Identify TBD of ticks collected during the COVID-19 pandemic in a rural community. Methods: Tick capture was carried out in March 2021, in Teabo, Yucatan. Ticks were removed using from domestic dogs and placed in ethanol. Collected ticks were morphologically identified and underwent DNA extraction and a partial segment of the mitochondrial 16S-rDNA gene was amplified to corroborate the tick species. The DNA was screened for the presence of Anaplasma spp., Borrelia spp., Ehrlichia spp., and Rickettsia spp. Purified amplification products were submitted for sequencing and the results were compared to those deposited in GenBank using BLAST. Results: We collected 33 ectoparasites, Ixodes affinis, Rhipicephalus sanguineus, Rhipicephalus microplus, and Amblyomma mixtum on 11 hunting dogs. The most frequent ectoparasite was R. sanguineus (66%). We detected the presence of DNA of Rickettsia endosymbiont in I. affinis and Anaplasma platys in R. sanguineus. Rickettsia endosymbiont presented a similarity of 100% with the partial sequence of R. endosymbiont of I. affinis isolate IACACTM001 16S ribosomal RNA gene and the sequence of A. platys had a similarity of 100% with the partial sequence of the isolate 23-33TX 16S ribosomal RNA gene of A. platys from dogs from Texas, USA and with the partial sequence of the isolate L134 16S ribosomal RNA gene of Ehrlichia canis from dogs from Piura, Peru. Conclusion: We confirmed for the first time the presence of A. platys in R. sanguineus and R. endosymbiont in I. affinis ticks from dogs in the state of Yucatan.

Diversity of Rickettsia spp. in ticks from wild mammals of Morocco and Mauritania.

Ticks are known as vectors and reservoirs of rickettsiae and, wildlife vertebrate hosts as suitable dispersers of ticks contributing to the life cycle of rickettsial agents in nature. In the herein study, the presence of rickettsiae was investigated in ticks from wild mammals (Gerbillus and Jaculus, Vulpes rueppellii, Canis anthus, Felis lybica and Felis margarita) in Mauritania and Morocco. Morphological and molecular analysis of ticks allowed their identification as Rhipicephalus sanguineus sensu lato and Hyalomma impeltatum. A total of 126 partially engorged adult ticks, collected from 40 animals, were screened for the presence of rickettsial DNA by conventional PCR targeting the ompB gene, followed by ompA and gltA targets and bidirectional sequencing. As a result of the sequence analyses, that at least three different species of pathogenic spotted fever group rickettsiae were detected. Rickettsia parkeri-like was detected in a R. sanguineus s.l. (n=1) collected from an African wildcat from Morocco. Rickettsia aeschlimannii was detected in a H. impeltatum (n=1) collected from a gerbil rodent. Rickettsia massiliae was detected in R. sanguineus s.l. ticks (n=5) collected from two Ruppells' foxes. The herein study demonstrates that pathogenic Rickettsia species are circulating in Morocco and Mauritania wildlife.

Molecular detection of tick-borne rickettsial pathogens in ticks collected from domestic animals from Cauca, Colombia.

Some hard ticks' species can act as vectors of a wide variety of pathogens of human and animal importance such as Anaplasma, Ehrlichia and Rickettsia spp. In Colombia, a total of forty-six tick species have been described, and some of them have been implicated as vectors of some infectious agents. The department of Cauca is one of the thirty-two departments of Colombia. Most of its population lives in rural areas and depends on agriculture as the main economic activity, favoring exposure to ticks and tick-borne pathogens. Thus, the present study aimed to determine the tick species and tick-borne pathogens circulating in this region. From August to November 2017, ticks were collected from dogs, horses and cattle from eight rural areas of four municipalities in the department of Cauca. All collected ticks were classified according to taxonomic keys and organized in pools. DNA was extracted from all tick pools for molecular confirmation of tick species and detection of Anaplasma, Ehrlichia and Rickettsia spp. A total of 2809 ticks were collected which were grouped in 602 pools. Ticks were morphologically identified as Amblyomma cajennense sensu lato, Dermacentor nitens, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato. The molecular identity of A. cajennense s.l. was confirmed as Amblyomma patinoi. A total of 95% of the pools scored positive for members of the Anaplasmataceae family, of which, 7.8% and 7.3% were positive to Anaplasma and Ehrlichia spp., respectively, being identified as Anaplasma marginale, Ehrlichia minasensis and Ehrlichia canis; and 16.1% were positive for Rickettsia spp. with high identity for Rickettsia asembonensis, Rickettsia felis and Candidatus Rickettsia senegalensis. This is the first report describing the natural infection of ticks with rickettsial pathogens and the occurrence of A. patinoi ticks in Cauca department, Colombia.

Tick-borne pathogens detected in sheltered dogs during an epidemic of Rocky Mountain spotted fever, a One Health challenge.

OBJECTIVE: To assess exposure to and infection with 3 pathogens (Rickettsia rickettsii, Anaplasma platys, and Ehrlichia canis) vectored by brown dog ticks (Rhipicephalus sanguineus) in sheltered dogs at the western US-Mexico border. ANIMALS: 239 dogs in shelters in San Diego and Imperial counties, US, and Mexicali and Tijuana, Mexico. PROCEDURES: Each dog had blood drawn and basic demographic data collected. PCR was performed to determine active infection with Rickettsia spp, E canis, and A platys. Serology was performed to determine exposure to Rickettsia, Anaplasma, and Ehrlichia species. RESULTS: 2 of 78 (2.6%) dogs sampled in Tijuana were actively infected with R rickettsii. A single brown dog tick collected from a dog in Tijuana was PCR-positive for R rickettsii. Infection with E canis and A platys ranged across shelters from 0% to 27% and 0% to 33%, respectively. Dogs in all 4 locations demonstrated exposure to all 3 pathogens, though Rickettsia and Ehrlichia seropositivity was highest in Mexicali (81% and 49%, respectively) and Anaplasma seropositivity was highest in Tijuana (45%). CLINICAL RELEVANCE: While infection and exposure were highest in sheltered dogs in the southern locations, dogs in all locations demonstrated exposure to all pathogens, demonstrating the potential for emergence and spread of zoonotic pathogens with significant public health consequences in southern California and northern Baja California. In addition, veterinarians and shelter staff should be aware that Ehrlichia or Anaplasma infection may co-occur with Rocky Mountain spotted fever, which is a human health risk.

First detection and molecular identification of Rickettsia massiliae, a human pathogen, in Rhipicephalus sanguineus ticks collected from Southern Taiwan.

The Rickettsia massiliae was firstly detected and identified in Rhipicephalus sanguineus ticks infested on dogs in Taiwan. A total of 1154 Rh. sanguineus ticks collected from 158 dogs of four districts of Tainan city were examined for Rickettsia infection by nested-PCR assay targeting the citrate synthase (gltA) and outer membrane protein B (ompB) genes of Rickettsia. The Rickettsia infection was detected with a general infection rate of 2.77%, and was detected in male, female and nymphal stage with an infection rate of 2.77%, 3.22% and 1.32%, respectively. Phylogenetic relationships were analyzed by comparing the gltA and ompB sequences obtained from 9 Taiwan strains and 16 other strains representing 13 genospecies of Rickettsia. Results revealed that all Taiwan strains were genetically affiliated to the same clades of R. massiliae (spotted fever group) and R. felis (transitional group), and can be discriminated from other genospecies of Rickettsia. This study provides the first evidence of R. massiliae, a pathogenic spotted fever Rickettsia, identified in Rh. sanguineus ticks and highlight the potential threat for the regional transmission of Rickettsia infection among humans in Taiwan.

Ehrlichia canis rapid spread and possible enzooty in northern South Australia and distribution of its vector Rhipicephalus linnaei.

Recent concerns have arisen in Australia regarding detections of the exotic bacterium Ehrlichia canis which has resulted in ehrlichiosis outbreaks. In Australia, it is spread by the tropical brown dog tick Rhipicephalus linnaei, formerly Rhipicephalus sanguineus sensu lato tropical lineage. Previously, the tick has been recorded in South Australia in the Coober Pedy and the Oodnadatta areas. This study, which includes historical specimens data held in historical Australian arthropod collections, along with 10 sampled remote communities, confirms the wide distribution range of this species within the State. E. canis was detected by PCR in the ticks. The percentage of dogs hosting PCR-positive ticks increased from 2.8% (95% confidence interval [CI]: 0.3 to 9.7) in November-December 2020 to 62.9% (95% CI: 44.9 to 78.5) end of February 2021, initially in two then in seven Anangu Pitjantjatjara Yankunytjatjara lands communities in the far northern regions of South Australia. Our results suggest a rapid spread of the pathogen. No evidence of E. canis was found in nine regional communities. The extended tropical brown dog tick distribution indicates a greater area where E. canis may occur and may require management to minimise the impacts of ehrlichiosis outbreaks. Without the implementation of effective detection and control programs, this extended distribution of R. linnaei is likely to result in the spread of the bacterium to other regions.

Use of a recombinant positive control in the diagnostic of canine Ehrlichiosis from 16sRNA gen of Ehrlichia canis in Mexico City.

Ehrlichia canis has gained importance over the years as a zoonotic bacterium, nevertheless in Mexico is unknown the extent of the problem in animals and public health. The country had a few studies carried out locally using serology and molecular tests as diagnostic methods. Ehrlichiosis is not considered endemic in the central valley of Mexico, because the climatic conditions in the region have not allowed the vector (Rhipicephalus sanguineus) to establish itself adequately, therefore, diagnosis is not used in clinical practice in this area. A nested PCR (nPCR) offers rapid results with high sensitivity and specificity regardless of cost. The use of a recombinant positive control provides the advantage of timely diagnosis, follow-up treatment and allows the clinician to decide. In this work, the nPCR reported by Wen et al. (J Clin Microbiol 35(7):1852-2185, 1997) was used for the diagnosis of E. canis by modifying the reaction conditions to improve the detection of the test. We constructed a recombinant positive control to nPCR as diagnostic technique for E. canis, also we modified the reaction conditions to improve detection of the test which allowed the diagnosis of E. canis in dogs in the Mexican Republic using 53 samples from dogs with positive serological diagnosis of Ehrlichiosis, some of them from the valley of Mexico. Currently, this nPCR is offered to public at the Faculty of Veterinary Medicine and Zootechnics of the National Autonomous University of Mexico at an accessible cost and allows to begin to generate epidemiological information to know distribution of the bacterium.