Bartonella spp. and Typhus Group Rickettsiae among Persons Experiencing Homelessness, São Paulo, Brazil.

Persons experiencing homelessness in São Paulo, Brazil, were seropositive for Bartonella spp. (79/109, 72.5%) and typhus group rickettsiae (40/109, 36.7%). Bartonella quintana DNA was detected in 17.1% (14/82) body louse pools and 0.9% (1/114) blood samples. Clinicians should consider vectorborne agents as potential causes of febrile syndromes in this population.

Rickettsia communities and their relationship with tick species within and around the national park of Iguaçu, Brazil.

We report Rickettsia species from 2,334 ticks collected from environment (1,939 ticks) and animals (395 ticks) in the largest inland fragment of the Atlantic rainforest of southern Brazil and its fragments. Additionally, the DNA infection rates of Amblyomma ovale tick populations in the Neotropics with Rickettsia bellii and Rickettsia parkeri strain Atlantic rainforest were calculated using data from scientific publications, and their correlation was evaluated. From 11 tick species Rickettsia DNA was detected in seven (Amblyomma brasiliense, Amblyomma coelebs, Amblyomma incisum, Amblyomma longirostre, A. ovale, Haemaphysalis juxtakochi, Ixodes fuscipes) and was not detected in four species (Amblyomma dubitatum, Ixodes loricatus, Rhipicephalus microplus and Rhipicephalus sanguineus sensu lato). DNA of five Rickettsia species was detected (R. bellii, Rickettsia amblyommatis, Rickettsia rhipicephali, Rickettsia felis and Rickettsia sp. Aragaoi). To determine the prevalence of Rickettsia DNA positivity according to vector species, ticks were processed individually or in pools of 2-10 individuals (samples). The most prevalent Rickettsia species was R. bellii, found in 112 samples, followed by R. amblyommatis, R. rhipicephali, R. felis and Rickettsia sp. Aragaoi, found in 16, five, two and one sample, respectively. Rickettsia bellii DNA was found in five tick species with the highest infection rate in A. ovale and A. brasiliense. Absence of R. parkeri strain Atlantic rainforest in A. ovale ticks was an unexpected result. Furthermore, a negative correlation was identified between the infection rates (DNA) of R. bellii and/or R. parkeri strain Atlantic rainforest within A. ovale tick populations in the Neotropics. Putting together current knowledge, it can be proposed that, within natural settings, the diversity of rickettsiae and ticks creates a buffering effect on the overgrowth of rickettsiae and episodes of bacteremia in the hosts.

Comparative genomics of the Western Hemisphere soft tick-borne relapsing fever borreliae highlights extensive plasmid diversity.

BACKGROUND: Tick-borne relapsing fever (TBRF) is a globally prevalent, yet under-studied vector-borne disease transmitted by soft and hard bodied ticks. While soft TBRF (sTBRF) spirochetes have been described for over a century, our understanding of the molecular mechanisms facilitating vector and host adaptation is poorly understood. This is due to the complexity of their small (~ 1.5 Mb) but fragmented genomes that typically consist of a linear chromosome and both linear and circular plasmids. A majority of sTBRF spirochete genomes' plasmid sequences are either missing or are deposited as unassembled sequences. Consequently, our goal was to generate complete, plasmid-resolved genomes for a comparative analysis of sTBRF species of the Western Hemisphere. RESULTS: Utilizing a Borrelia specific pipeline, genomes of sTBRF spirochetes from the Western Hemisphere were sequenced and assembled using a combination of short- and long-read sequencing technologies. Included in the analysis were the two recently isolated species from Central and South America, Borrelia puertoricensis n. sp. and Borrelia venezuelensis, respectively. Plasmid analyses identified diverse sequences that clustered plasmids into 30 families; however, only three families were conserved and syntenic across all species. We also compared two species, B. venezuelensis and Borrelia turicatae, which were isolated ~ 6,800 km apart and from different tick vector species but were previously reported to be genetically similar. CONCLUSIONS: To truly understand the biological differences observed between species of TBRF spirochetes, complete chromosome and plasmid sequences are needed. This comparative genomic analysis highlights high chromosomal synteny across the species yet diverse plasmid composition. This was particularly true for B. turicatae and B. venezuelensis, which had high average nucleotide identity yet extensive plasmid diversity. These findings are foundational for future endeavors to evaluate the role of plasmids in vector and host adaptation.

Novel Anaplasmataceae agents Candidatus Ehrlichia hydrochoerus and Anaplasma spp. Infecting Capybaras, Brazil.

We amplified Ehrlichia and Anaplasma DNA from Amblyomma dubitatum tick-infested capybaras (Hydrochoerus hydrochaeris) in southern Brazil. Sequencing of 16S rRNA, sodB, and groEL indicated a novel Ehrlichia species, and sequencing of 16S rRNA from 2 capybaras indicated a novel Anaplasma species. The tick vectors remain unknown.

Rickettsial infection in ticks from a natural area of Atlantic Forest biome in southern Brazil.

From June 2013 to January 2014, blood sera samples and ticks were collected from domestic dogs and wild small mammals, and ticks from the vegetation in a preservation area of the Atlantic Forest biome (Turvo State Park), and the rural area surrounding the Park in Derrubadas municipality, state of Rio Grande do Sul, southern Brazil. Dogs were infested by Amblyomma ovale and Amblyomma aureolatum adult ticks, whereas small mammals were infested by immature stages of A. ovale, Amblyomma yucumense, Amblyomma brasiliense, Ixodes loricatus, and adults of I. loricatus. Ticks collected on vegetation were A. brasiliense, A. ovale, A. yucumense, Amblyomma incisum, and Haemaphysalis juxtakochi. Three Rickettsia species were molecularly detected in ticks: Rickettsia bellii in I. loricatus (also isolated through cell culture inoculation), Rickettsia amblyommatis in A. brasiliense, and Rickettsia rhipicephali in A. yucumense. The latter two are tick-rickettsia associations reported for the first time. Seroreactivity to Rickettsia antigens were detected in 33.5% (55/164) small mammals and 8.3% (3/36) canine sera. The present study reveals a richness of ticks and associated-rickettsiae in the largest Atlantic Forest Reserve of the state of Rio Grande do Sul, which is characterized by a rich fauna of wild mammals, typical of more preserved areas of this biome. Noteworthy, none of the detected Rickettsia species have been associated to human or animal diseases. This result contrasts to other areas of this biome in Brazil, which are endemic for tick-borne spotted fever caused by Rickettsia rickettsii or Rickettsia parkeri.

Serosurvey of spotted fever group Rickettsia in equids from western Pará, Amazon, Brazil.

From a previous large epidemiological survey, we randomly selected 474 serum samples (463 horses and 11 mules) distributed among four municipalities of Pará state, Amazon region, Brazil, and from three types: farm animal, urban carthorse, and sport horse. Samples were tested by indirect fluorescence antibody test (IFAT ≥ 64) for antibodies reactive to spotted fever group (SFG) rickettsiae using Rickettsia rickettsii as crude antigens. From the 474 equids tested, 149 (31.4%) had ticks attached during sampling, belonging mostly to the species Dermacentor nitens. The overall seroprevalence for SFG rickettsiae was 31.4% (95% confidence interval: 27.3-35.9%) with 149 seropositive animals out of 474 screened. Notably, 77 equids (16.2%) had high endpoint titers ranging from 512 to 16,384, indicating that they had been exposed to SFG rickettsiae not long before sampling. Animal type affected rickettsial seroprevalence, with significantly higher values among farm horses when compared with urban and sport animals. Presence of dogs and tick infestation were negatively associated with equid seropositivity to R. rickettsii. This is the first report of SFG rickettsiae-reactive antibodies in equids from Pará state, Brazilian Amazon.

New records of soft ticks (Acari: Argasidae) in the Caatinga biome of Brazil, with a phylogenetic analysis of argasids using the nuclear Histone 3 (H3) gene.

In Brazil, 19 species of the genus Ornithodoros (Acari: Argasidae) have been reported. The medical and veterinary importance of Ornithodoros ticks has increased substantially in recent decades, with the discovery of various relapsing fever Borrelia infecting Ornithodoros ticks. Herein, argasid ticks were collected during 2019-2020 from caves, abandoned nests and homes in various regions of Ceará State, Brazilian semiarid-Caatinga biome. In total, 289 ticks were collected and identified into five species: Ornithodoros cavernicolous (176 specimens), Ornithodoros fonsecai (81), Ornithodoros mimon (12), Ornithodoros rietcorreai (4), and a fifth species provisionally retained as Ornithodoros sp. Ubajara. Tick identifications were corroborated by a phylogenetic analysis inferred using the 16S rRNA gene. To extend the molecular characterization, DNA samples were tested by an additional PCR assay targeting the nuclear Histone 3 (H3) gene. Because there were no H3 sequences of argasids in GenBank, we extended this PCR assay for additional Ornithodoros species, available in our laboratory. In total, 15 partial sequences of the H3 gene were generated for 10 Ornithodoros species, showing 0% intraspecific polymorphism, and 1.5-11.6% interspecific polymorphism. Phylogenetic analyses inferred segregated Ornithodoros sp. Ubajara as a potential novel species. Our results also highlight the potential of the H3 gene for deeper phylogenetic analyses of argasids. The present study provides new data for argasid ticks of the genus Ornithodoros in the Caatinga biome. Because some of these tick species are human-biting ticks, active surveillance for the incidence of human infection due to Ornithodoros-borne agents is imperative in the Caatinga biome.

Relapsing Fever Group Borreliae in Human-Biting Soft Ticks, Brazil.

We conducted a molecular survey for Borrelia spp. in Ornithodoros ticks previously reported as biting humans. We collected specimens in natural ecosystems and inside human dwellings in 6 states in Brazil. Phylogenetic analyses unveiled the occurrence of 4 putatively new species of relapsing fever group borreliae.

Life cycle of Ixodes schulzei (Acari: Ixodidae) in the laboratory, and demonstration of reproduction by parthenogenesis.

Ixodes schulzei is an ixodid tick that parasitizes Cricetidae rodents, chiefly the South American water rat, Nectomys squamipes, in Brazil and Argentina. In the present study, we evaluated the life cycle of I. schulzei by exposing larvae and nymphs to feed on two rodent species, N. squamipes and Calomys callosus (large vesper mouse),while adult ticks were exposed to feed on N. squamipes. Off-host developmental periods were observed in an incubator at 27 °C, 95% relative humidity, and 0:24 (light:dark) regimen. Larvae and nymphs successfully fed on either C. callosus or N. squamipes. Mean larval and nymphal feeding periods were 8.8 and 8.7 days on N. squamipes and 8.5 and 9.7 days on C. callosus. The majority of engorged larvae (79.0-80.8%) and nymphs (67.0-86.0%) successfully molted to nymphs and adults, respectively. Mean premolt periods were 11.5-11.7 days for engorged larvae and 22.5-23.7 days for engorged nymphs. Only adult females emerged from engorged nymphs, regardless of host species, i.e., none of 120 engorged nymphs molted to male. Around 18% of the unfed females presented teratologies compatible with the metagynander type of gynandromorphism. Ixodes schulzei adult females successfully fed (mean feeding period, 9.4 days), oviposited, and presented high reproductive performance (high engorged weight, egg mass weight, and % egg mass hatching), in the absence of male ticks. Our results showed that I. schulzei successfully reproduces by parthenogenesis, and corroborate field data that indicate N. squamipes as the most important host for this tick species. The male of I. schulzei remains unknown.

Distribution modeling of Amblyomma rotundatum and Amblyomma dissimile in Brazil: estimates of environmental suitability.

The number of reports of tick parasitism in amphibians and reptiles has increased over the past few years, including new host and location records for Amblyomma rotundatum and Amblyomma dissimile. However, knowledge of the geographical distribution remains incomplete, and in many regions of Brazil, the presence of these vectors has not been investigated. Several candidate models were built using a correlative maximum entropy approach, and best-fitting models were selected based on statistical significance, predictive ability, and complexity based on current climatic trends and future projected climate changes. Final models showed a good ability to discriminate A. rotundatum and A. dissimile current and future potential distributions. The entire country had higher predicted suitability for A. rotundatum while A. dissimile was mainly restricted to the Amazon and Pantanal biomes. A. rotundatum is a species with enormous potential for dissemination in the next decades, potentially through the legal and illegal transport of reptiles and amphibians. The proposed model is useful for targeting surveillance efforts increasing the efficiency and accuracy of future ecological research and tick management efforts.

Ticks and tick-borne Rickettsia in El Salvador.

From May to November 2013, ticks were collected from wild and domestic hosts that were sampled by convenience in different localities of El Salvador. Among 48 localities, in total 1181 ticks were collected from 200 vertebrate animals, comprising 13 species of wild hosts (amphibian, reptiles, mammals) and five species of domestic mammals, plus four samples from humans and four samples from the environment. Through morphological analysis (corroborated by molecular analyses in a few cases), the following ten tick species were identified: Amblyomma dissimile, Amblyomma mixtum, Amblyomma ovale, Amblyomma cf. parvum, Amblyomma sabanerae, Amblyomma scutatum, Dermacentor dissimilis, Dermacentor nitens, Rhipicephalus microplus, and Rhipicephalus sanguineus sensu lato. Among a sample of 211 tick specimens tested for rickettsial infection by molecular methods, we identified: 'Candidatus Rickettsia colombianensi' in 10% of the A. dissimile ticks and 11% of the A. scutatum ticks; Rickettsia amblyommatis in 77% of the A. mixtum ticks, 50% of the A. cf. parvum ticks, 8% of the D. nitens ticks, and 11% of the Amblyomma spp. nymphs; and Rickettsia bellii in 3% of the A. dissimile ticks and 17% of the A. ovale ticks. The tick fauna of El Salvador is currently represented by 12 reported species.

Morphological descriptions of the nymph and adults of Ornithodoros clarki, the larva and nymph of Ornithodoros rondoniensis, with notes on their phylogenetic relationships.

Based on tick specimens collected recently in Mexico, Nicaragua, Panama and Brazil, we provide morphological descriptions of the nymph and adults of Ornithodoros clarki Jones & Clifford, 1972 from the first three countries, and the larva and nymph of Ornithodoros rondoniensis (Labruna, Terassini, Camargo, Brandão, Ribeiro & Estrada-Peña, 2008) from Brazil. Also, an analysis of mitochondrial 16S rDNA sequences was performed to analyze the phylogenetic relationships of these tick species. Adults and nymphs of O. clarki and O. rondoniensis are unique among the Argasidae family by presenting exceptionally large spiracular plates with small goblets, and an integument with smooth polygonal mammillae. However, these two species are morphologically distinct based on specific patterns of coxal folds, idiosomal mammillae and pilosity, and female genital flap. In contrast, the larvae of O. clarki and O. rondoniensis are morphologically identical, except for a general larger size of the former species; this slight difference is corroborated by Principal Component Analysis (PCA) by using 40 morphometric variables. Phylogenetic analyses including 16S rDNA partial sequences of different Ornithodoros taxa from Central and South America indicate that O. rondoniensis from Brazil diverges from O. clarki from Mexico, Nicaragua and Panama. However, phylogenetic distance separating both alleged species is similar or slightly lower than the distances depicted for conspecific populations of a few other Ornithodoros species. Nonetheless, our primary criterion to maintain O. rondoniensis as a valid species is because its adult and nymphal stages do present distinct morphological traits that easily distinguish these postlarval stages from O. clarki.

Tick-borne zoonotic agents infecting horses from an urban area in Midwestern Brazil: epidemiological and hematological features.

The emergence of tick-borne diseases has been reported as a serious problem in public health worldwide and many aspects of its epidemiology and effects on the health of its hosts are unclear. We aimed to perform an epidemiological study of tick-borne zoonotic Rickettsia, Borrelia, and Anaplasmataceae in horses from Midwestern Brazil. We also evaluated whether Borrelia spp. and Anaplasmataceae may be associated with hematological disorders in the sampled animals. Blood and serum samples as well as ticks were collected from 262 horses. Serum samples were used to perform serological tests, and hematological analyses were made using whole blood. Furthermore, DNA extracted from whole blood and ticks was used for molecular tests. Campo Grande is enzootic for tick-borne studied bacteria, since we found an overall exposure of 59.9% of the sampled horses, 28.7% of them presented co-exposure. Seropositivity rates of 20.6% for Borrelia spp., 25.6% for Rickettsia spp., and 31.6% for Anaplasmataceae were found in the sampled horses. Considering both molecular and serological tests for Borrelia spp., the infection rate was 48.0% (126/262). None of the tested horses showed molecular positivity for Anaplasma phagocytophilum. The horses sampled displayed 7.2% of parasitism by ixodid ticks in single and coinfestations. We did not find DNA of any studied bacteria in the sampled ticks. Positive horses for Borrelia spp. and Anaplasmataceae agents displayed leukopenia, monocytopenia, and lymphopenia. Together, our results suggest that horses may play a role as sentinel host for zoonotic bacteria and Borrelia spp. and Anaplasmataceae agents can impair the health of horses.

Typhus Group Rickettsiosis, Brazilian Amazon.

Rickettsia rickettsii infection is the only rickettsiosis included in the list of reportable diseases in Brazil, where typhus group rickettsioses, mainly murine typhus, have been underreported. We report a case of typhus group rickettsiosis with unique ecologic particularities in a patient from the Brazilian Amazon, where, to our knowledge, rickettsioses have not been reported.

Needlestick-Associated Rocky Mountain Spotted Fever, Brazil.

We report a fatal case of Rocky Mountain spotted fever (RMSF) in a man in Brazil without recent history of tick bites or environmental exposure. He received an accidental needlestick while working as a nurse. The nurse and his patient died. Both cases were confirmed as RMSF by molecular methods.

"Candidatus Borrelia ibitipoquensis," a Borrelia valaisiana-Related Genospecies Characterized from Ixodes paranaensis in Brazil.

Borrelia burgdorferi sensu lato (Bbsl) spirochetes include the agents of Lyme borreliosis in temperate regions of the Northern Hemisphere, and merge their transmission cycles mainly with ticks of the Ixodes ricinus complex. Twenty genospecies compose Bbsl currently, and with the exception of Borrelia chilensis, and Borrelia garinii, all have been described only for North America, Europe, North Africa, and Asia. Here, we collected specimens of Ixodes paranaensis, a tick associated with swifts in a Brazilian natural park from the state of Minas Gerais, and performed a molecular characterization of 11 borrelial genes. Based on comparisons of inter and intraspecific genetic divergences, and Bayesian phylogenetic trees inferred for 16S rRNA, flaB, p66, and concatenated clpA, clpX, pepX, pyrG, recG, nifS rlpB, and uvrA genes, we demonstrate the occurrence of a new genospecies of Bbsl. "Candidatus Borrelia ibitipoquensis" Ip37 is closely related to Borrelia sp. Am501, and Borrelia valaisiana, a spirochete transmitted by ticks of the I. ricinus complex in Eurasia that uses birds as reservoirs. In a similar ecological scenario involving ticks and avian hosts, the migratory swift Streptoprocne biscutata is the sole-documented bird associated with I. paranaensis, and, although not assessed in this study, could correspond to the vertebrate reservoir of this newly described genospecies in Brazil. Pathogenic roles of "Ca. B. ibitipoquensis" are still unknown. However, its possible vector I. paranaensis is not an anthropophilic tick, so human infections would be unlikely to occur. Our finding enhances the knowledge on Bbsl in South America, highlights the occurrence of ecologically and genetically related genospecies with vastly separated geographical distributions, and calls for the attention to explore a barely known diversity of spirochetes of this group in the region.

Low host specificity and lack of parasite avoidance by immature ticks in Brazilian birds.

Ticks are ectoparasites that feed on blood of a broad taxonomic range of terrestrial and flying vertebrates and are distributed across a wide range of environmental conditions. Here, we explore the biotic and abiotic factors on infestation probability of ticks of the genus Amblyomma and assess the degree of host specificity based on analysis of 1028 birds surveyed across Brazil. We show that tick infestation rates exhibited considerable variation across the 235 avian species analyzed and that the probability of an individual bird being parasitized by immature ticks (larvae and nymphs) increased with annual precipitation. Host phylogeny and two host ecological traits known to promote tick exposure (body mass and foraging behavior) did not predict infestation probability. Moreover, immature ticks displayed a low degree of host specificity at the family level. Lastly, tick occurrence in birds carrying infection with avian malaria and related parasites did not differ from those free of these haemosporidian parasites, indicating a lack of parasite avoidance by immature ticks. Our findings demonstrate that tick occurrence in birds across Brazilian biomes responds to environmental factors rather than ecological and evolutionary host attributes.

Ornithodoros capensis sensu stricto (Ixodida: Argasidae) in Coiba National Park: first report for Panama, with notes on the O. capensis group in Panamanian shores and Costa Rica.

Ornithodoros capensis sensu lato (s.l.) is a morphologically similar group of soft ticks that parasitizes mostly seabirds in continental and offshore territories worldwide. Ornithodoros capensis sensu stricto (s.s.) has been previously recorded in many islands and coastal localities along the American continent; however, some records from Central America remain obscure. In this work we performed morphological and molecular analyses on soft ticks collected in Coiba National Park, an archipelago located in the Pacific Ocean off the coast of Panama, confirming the occurrence of O. capensis s.s. in this country for the first time. In addition, a morphological examination of museum specimens collected in Costa Rica, and a further locality in Panama, confirmed that O. capensis s.l. is established in the former country, and that its distribution along Panamanian shores is likely larger.

Maintenance of the infection by Rickettsia amblyommatis in Amblyomma cajennense sensu stricto ticks and evaluation of vector competence.

Rickettsia amblyommatis has been reported infecting various tick species throughout the western hemisphere, including Amblyomma cajennense sensu stricto (s.s.). The present study aimed to evaluate whether R. amblyommatis can be maintained by transovarial transmission and transstadial passage in A. cajennense s.s. ticks, and whether these ticks are competent vectors of this rickettsial agent. We selected engorged females that were naturally infected or uninfected by R. amblyommatis, and reared their offspring (infected and control groups, respectively). Immature ticks were allowed to feed on guinea pigs, whereas adults fed on rabbits. All stages and individuals of each generation of the infected group yielded rickettsial DNA, which was not detected in any tick from the control group. No host from the control group seroconverted to R. amblyommatis. Among 16 guinea pigs and eight rabbits infested with R. amblyommatis-infected ticks, only one guinea pig seroconverted to R. amblyommatis. Some unfed adult ticks of the infected group were dissected, and DNA was extracted from their salivary glands and from their carcasses. DNA of R. amblyommatis was detected in all carcasses, but not in the salivary glands. Results of this study indicate very low vector competence of A. cajennense s.s. for R. amblyommatis. Previous studies reported domestic animals with high titers to R. amblyommatis in areas where these animals are naturally infested chiefly by A. cajennense s.s. ticks. One may consider that the vector competence of A. cajennense s.s. for R. amblyommatis could vary among tick populations and/or rickettsial strains.

Host movement and time of year influence tick parasitism in Pantanal birds.

Ticks are among the best studied parasitic groups as they spread important pathogens of medical and veterinary importance worldwide. Migratory birds can play an important role in transporting ticks infected with pathogens across wide geographic regions. It is therefore important to understand which factors promote tick parasitism rates across their avian hosts and the associated potential for disease spread. Here, we identified the host attributes of infestation probability of ticks from the genus Amblyomma in 955 birds from Pantanal, Brazil. Infestation rates exhibited considerable variation across the 129 avian species surveyed and were explained by both host ecological traits and evolutionary history. The probability of an individual bird being infested with immature ticks (larvae and/or nymphs) was higher across resident bird species that forage at ground level and during the wet season. Bird species that feed on vertebrates were less likely to be infested by ticks. Other ecological traits known to promote tick exposure (age, body mass, social behavior, and sex) did not predict infestation probability. Our findings demonstrate that tick occurrence in Pantanal birds is determined by avian host attributes, but tick occurrence throughout the year constrains exposure to host-seeking ticks. Moreover, the ecology of the avian host might prevent the potential spread of tick-borne diseases outside Pantanal as migratory hosts are likely less infested by ticks.