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Tick-borne relapsing fever spirochetes of genus Borrelia thrive in enzootic cycles involving Ornithodoros spp. (Argasidae) mainly, and rodents. The isolation of these spirochetes usually involves a murine model in which ticks are fed and the spirochetes detected in blood several days later. Such an experiment also demonstrates that a given species of tick is competent in the transmission of the bacteria. Here, soft ticks Ornithodoros octodontus were collected in Northern Chile with the objective to experimentally determine its capacity to transmit a Borrelia sp. detected in a previous study. Two Guinea pigs (Cavia porcellus) were used to feed nymphs and adults of O. octodontus and the spirochetes in blood were inspected by dark-field microscopy and nested PCR. Although spirochetes were not seen in blood, DNA was detected in only one animal 11 days after the ticks were fed. Genetic sequences of Borrelia flaB, clpX, pepX, recG, rplB, and uvrA genes retrieved from DNA extraction of positive blood were employed to construct two phylogenetic analyses. On the one hand, the flaB tree showed the Borrelia sp. transmitted by O. octodontus clustering with Borrelia sp. Alcohuaz, which was previously detected in that same tick species. On the other hand, concatenated clpX-pepX-recG-rplB-uvrA demonstrated that the characterized spirochete branches together with "Candidatus Borrelia caatinga", a recently discovered species from Brazil. Based on the genetic profile presented in this study, the name "Candidatus Borrelia octodonta" is proposed for the species transmitted by O. octodontus. The fact that spirochetes were not observed in blood of guinea pigs, may reflect the occurrence of low spirochetemia, which could be explained because the susceptibility of infection varies depending on the rodent species that is used in experimental models. Although the vertebrate reservoir of "Ca. Borrelia octodonta" is still unknown, Octodon degus, a rodent species that is commonly parasitized by O. octodontus, should be a future target to elucidate this issue.

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Bats represent the second order of mammals with the highest number of species worldwide with over 1,616 species, and almost 10% of them are recorded in Mexico. These mammals have a great diversity of ectoparasites, in particular soft ticks of the genus Ornithodoros. Desmodus rotundus is one of the bat species that has scarcely been studied in terms of tick species richness in Mexico, with three tick species reported in five of the 32 Mexican states. For this reason, the aim of the present work was to identify ticks associated with D. rotundus from Central Mexico. Fieldwork was undertaken in the municipality El Marqués, Ejido Atongo A, Querétaro, Mexico. Bats were captured using mist nets and were visually inspected for tick presence. The ectoparasites were identified morphologically and molecularly with the use of mitochondrial markers 16SrDNA and cytochrome oxidase subunit I (COI). A total of 30 D. rotundus (1 female, 29 males) were captured, from which 20 larvae identified as Ornithodoros yumatensis were recovered. Molecular analysis confirmed the presence of this species with identity values of 99-100% with sequences of this species from the southwestern US, and the Yucatán Peninsula, Mexico. This is the first report of ticks associated with bats for the state of Querétaro, providing the first sequences of the COI gene from Mexican populations of O. yumatensis and shows an increase in the distribution of this soft tick across Central Mexico.

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Soft ticks (Argasidae) of the Pavlovskyella Pospelova-Shtrom subgenus are important vectors of relapsing fever spirochetes, which are agents of disease globally. South American representatives of the Pavlovskyella subgenus include 3 species: Ornithodoros (Pavlovskyella) brasiliensis Aragão, Ornithodoros (Pavlovskyella) furcosus Neumann, and Ornithodoros (Pavlovskyella) rostratus Aragão. Here, we describe a fourth species based on morphological and mitogenomic evidence of ticks collected in burrows of unknown hosts in central Chile. The larva of the new species separates from other South American soft ticks by the following combination of characters: 13 pairs of dorsolateral setae, dorsal plate hexagonal, hypostome blunt with denticles from apex almost to the base. Adults of this new species lack cheeks, possess a dorsoventral groove, and have humps, similar to O. (P.) brasiliensis; however, they lack bulging structures on the flanks of idiosoma. Moreover, females and males differ from O. (P.) rostratus by having 3 humps instead of spurs in tarsi I and from O. (P.) furcosus because of their smaller size and thinner anterior lip of the genital aperture in females. The phylogenetic analysis performed with mitogenomes of the Argasidae family depicts the new Pavlovskyella species from Chile in a monophyletic clade with other South American species in the subgenus, confirming a regional group.

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Soft ticks are neglected competent vectors of a wide range of pathogenic microorganisms, among which bacteria of the genera Rickettsia and Borrelia stand out. In Mexico, previous studies have shown the presence of a member of the Ornithodoros talaje complex in the Virginia opossum (Didelphis virginiana Didelphimorphia: Didelphidae Kerr) from southeastern Mexico. However, its specific identification has not been achieved. Two D. virginiana were treated in a private clinic during the period of April-May 2022. Tick larvae were manually removed, DNA extraction was performed, and some genes from various bacterial and parasitic pathogens were amplified and sequenced. A total of 96 larvae were recovered, which were morphologically identified as Ornithodoros puertoricensis (Ixodida: Argasidae Fox); the 16 S sequences showed a similarity of 96.79%-99.51% with sequences of O. puertoricensis from Panama and Colombia. The presence of Rickettsia felis (Rickettsiales: Rickettsiaceae Bouyer et al.) was detected in 15 specimens from one host. The soft tick O. puertoricensis is recorded for the first time as an ectoparasite of the Virginia opossum in America and represents the second report for this soft tick in Mexico since 1963. This represents the most northern record of this tick species in its geographic distribution and brings a new soft tick-Rickettsia association.

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Ornithodoros mimon is an argasid tick species usually associated with bats and marsupials and occasionally parasitizes humans inside their homes. This paper reports a tick infestation in a residence in the municipality of Campinas, located in the interior of the state of São Paulo (SP). This report increases O. mimon occurrence in SP and corroborates its anthropophilic activity. Further studies are needed to clarify its role as a vector of pathogens. We highlighted the presence of O. mimon in an area with a large human population (Campinas) associated with synanthropic animals.(AU)

Ornithodoros mimon é uma espécie de carrapato argasídeo, geralmente associada a morcegos e marsupiais, sendo ocasionalmente relatada parasitando humanos dentro de seus domicílios. Este trabalho relata a infestação por carrapatos em uma residência no município de Campinas, interior do estado de São Paulo (SP). O presente relato amplia a ocorrência de O. mimon no estado de SP, corroborando sua atividade antropofílica, sendo necessários mais estudos para esclarecer o seu possível papel como vetor de patógenos. Destaca-se a presença de O. mimon numa área de grande contingente humano (Campinas), associado a animais sinantrópicos.(AU)

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Interest in research on soft ticks has increased in recent decades, leading to valuable insight into their role as disease vectors. The use of metagenomics-based analyses have helped to elucidate ecological factors involved in pathogen, vector, and host dynamics. To understand the main bacterial assemblages present in Ornithodoros cf. hasei and its mammalian hosts, 84 ticks and 13 blood samples from bat hosts (Chiroptera) were selected, and the 16S rRNA gene V4 region was sequenced in five pools (each one related to each host-tick pairing). Bacterial taxonomic assignment analyses were performed by comparing operational taxonomic units (OTUs) shared between ticks and their host blood. This analysis showed the presence of Proteobacteria (38.8%), Enterobacteriaceae (25%), Firmicutes (12.3%), and Actinobacteria (10.9%) within blood samples, and Rickettsiaceae (39%), Firmicutes (25%), Actinobacteria (13.1%), and Proteobacteria (9%) within ticks. Species related to potentially pathogenic genera were detected in ticks, such as Borrelia sp., Bartonella tamiae, Ehrlichia sp. and Rickettsia-like endosymbiont, and the presence of these organisms was found in all analyzed bat species (Cynomops planirostris, Molossus pretiosus, Noctilio albiventris), and O. cf. hasei. About 41-48.6% of bacterial OTUs (genera and species) were shared between ticks and the blood of bat hosts. Targeted metagenomic screening techniques allowed the detection of tick-associated pathogens for O. cf. hasei and small mammals for the first time, enabling future research on many of these pathogens.

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Ornithodoros tabajara n. sp. is described from laboratory-reared larvae and adult specimens collected in the Brazilian Caatinga. This new species shares the ecological niche with Ornithodoros rietcorreai and is likely associated with colonial rodents of genus Kerodon. However, O. tabajara n. sp. is morphologically easy to distinguish from O. rietcorreai and other Neotropical Ornithodoros by a unique combination of characters: larva with 17 pairs of dorsal setae (seven anterolateral, three central and seven posterolateral), sub-oval dorsal plate, hypostome blunt apically with dentition formula 2/2 along its extension, only one pair of posthypostomal setae, six pairs of sternal setae, posteromedian setae absent, and leave-shaped anal valves; alive adults with whitish islands of mammillae symmetrically distributed on dorsum (not visible in ethanol-preserved specimens), and median disk merging with posteromedian file. A phylogenetic analysis performed with mitochondrial 16S rDNA sequences points O. tabajara n. sp. as O. rietcorreai's sister taxon, which rises the hypothesis of sympatric speciation.

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Resumen Este informe describe el hallazgo de garrapatas blandas (Ixodida, Argasidae) parasitando a dos carpinteros andinos (Colaptes rupicola) provenientes del distrito de Nuñoa, provincia de Melgar en Puno, Perú (14°31'11.77"S; 70°32'15.95"W; 3967 m de altitud). Un total de 29 larvas de garrapatas fueron colectadas directamente de las aves y posteriormente identificadas morfológicamente como Argas neghmei. El diagnóstico fue basado en la morfología de las larvas y confirmado mediante el análisis molecular del gen 16S ARNr mitocondrial de la garrapata. Un fragmento de 363 pares de bases de este gen mostró una identidad del 100% con secuencias registradas para A. neghmei de Argentina y Chile (GenBank: FJ853598 y DQ295781). Este hallazgo confirma la presencia de A. neghmei en Perú y agrega al carpintero andino como nuevo hospedero para esta garrapata.

Abstract This report describes the finding of soft ticks (Ixodida, Argasidae) parasitizing two Andean flickers (Colaptes rupicola) from the Nuñoa district, Melgar province in Puno, Peru (14°31'11.77"S; 70°32'15.95''W; elevation 3967 m). A total of 29 larval ticks were collected directly from the birds and morphologically identified as Argas neghmei. Diagnosis was based on the larval morphology and confirmed by molecular analysis of the tick mitochondrial 16S rRNA gene. A sequence of 363 base pair of this gene showed to be 100% identical with sequences of A. neghmei from Argentina and Chile (GenBank: FJ853598 and DQ295781). This finding confirms the presence of A. neghmei in Peru and adds the Andean flicker as a new host for this species.

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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.

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Ornithodoros fonsecai is an argasid tick that is endemic to Brazil and has been described in the municipality of Bonito, state of Mato Grosso do Sul. Some specimens of this species were found in a cave in the municipality of Nobres, state of Mato Grosso. The specific identification of this population was confirmed by means of morphology and molecular biology. The mitochondrial 16S rDNA partial sequence of this species from Nobres has been deposited in GenBank (MK158949). The objective of this study was to elucidate the biology of O. fonsecai from Nobres, and to report autogeny in this tick population. Along three laboratory generations was observed molting of first nymphal instar to the second instar without feeding, a typical behavior of species included in the subgenus Alectorobius. The first generation (F1) presented five nymphal instars (N1 to N5), and most of adults emerged through molting of N5. The last nymphal instar of second generation (F2) was N4, but most of adults emerged from N3. In the third generation (F3) the last nymphal instar was N5, with most of the adults emerging from N4. In F2, some females (n = 20) originated from N3 began laying eggs without a blood meal. It was observed that those N3 fed twice before they molted to autogenic females. However, autogenic behavior occurred in relation to third generation females (F3) with specimens originating from N4 (n = 12) that were fed only once as nymphs. This behavior has already been reported as obligatory for the genera Otobius and Antricola, while it is facultative for one species of genus Argas and for four species of genus Ornithodoros. However, the present report provides the first record of facultative autogeny for a species of Ornithodoros in Brazil.

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BACKGROUND: Despite the increasingly recognized eco-epidemiological significance of bats, data from molecular analyses of vector-borne bacteria in bat ectoparasites are lacking from several regions of the Old and New Worlds. METHODS: During this study, six species of ticks (630 specimens) were collected from bats in Hungary, Romania, Italy, Kenya, South Africa, China, Vietnam and Mexico. DNA was extracted from these ticks and analyzed for vector-borne bacteria with real-time PCRs (screening), as well as conventional PCRs and sequencing (for pathogen identification), based on the amplification of various genetic markers. RESULTS: In the screening assays, Rickettsia DNA was only detected in bat soft ticks, whereas Anaplasma phagocytophilum and haemoplasma DNA were present exclusively in hard ticks. Bartonella DNA was significantly more frequently amplified from hard ticks than from soft ticks of bats. In addition to Rickettsia helvetica detected by a species-specific PCR, sequencing identified four Rickettsia species in soft ticks, including a Rickettsia africae-like genotype (in association with a bat species, which is not known to migrate to Africa), three haemotropic Mycoplasma genotypes in Ixodes simplex, and Bartonella genotypes in I. ariadnae and I. vespertilionis. CONCLUSIONS: Rickettsiae (from both the spotted fever and the R. felis groups) appear to be associated with soft rather than hard ticks of bats, as opposed to bartonellae. Two tick-borne zoonotic pathogens (R. helvetica and A. phagocytophilum) have been detected for the first time in bat ticks. The present findings add Asia (China) to the geographical range of R. lusitaniae, as well as indicate the occurrence of R. hoogstraalii in South Africa. This is also the first molecular evidence for the autochthonous occurrence of a R. africae-like genotype in Europe. Bat haemoplasmas, which are closely related to haemoplasmas previously identified in bats in Spain and to "Candidatus Mycoplasma haemohominis", are reported here for the first time from Central Europe and from any bat tick.

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Ornithodoros fonsecai is an argasid tick that is endemic to Brazil and has been described in the municipality of Bonito, state of Mato Grosso do Sul. Some specimens of this species were found in a cave in the municipality of Nobres, state of Mato Grosso. The specific identification of this population was confirmed by means of morphology and molecular biology. The mitochondrial 16S rDNA partial sequence of this species from Nobres has been deposited in GenBank (MK158949). The objective of this study was to elucidate the biology of O. fonsecai from Nobres, and to report autogeny in this tick population. Along three laboratory generations was observed molting of first nymphal instar to the second instar without feeding, a typical behavior of species included in the subgenus Alectorobius. The first generation (F1) presented five nymphal instars (N1 to N5), and most of adults emerged through molting of N5. The last nymphal instar of second generation (F2) was N4, but most of adults emerged from N3. In the third generation (F3) the last nymphal instar was N5, with most of the adults emerging from N4. In F2, some females (n?=?20) originated from N3 began laying eggs without a blood meal. It was observed that those N3 fed twice before they molted to autogenic females. However, autogenic behavior occurred in relation to third generation females (F3) with specimens originating from N4 (n?=?12) that were fed only once as nymphs. This behavior has already been reported as obligatory for the genera Otobius and Antricola, while it is facultative for one species of genus Argas and for four species of genus Ornithodoros. However, the present report provides the first record of facultative autogeny for a species of Ornithodoros in Brazil.

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ORNITHODOROS BRASILIENSIS: Aragão is an endemic tick restricted to the highlands of the state of Rio Grande do Sul, Brazil. This species is very aggressive toward humans, causing fever, great pain and intense inflammatory response at the bite site. It is also very aggressive toward dogs, and tick toxicosis syndrome has been reported in this host. In order to elucidate the biology of this tick, the present study describes its life cycle under laboratory conditions, using guinea pigs as hosts for two generations. In the nonparasitic phase, the ticks were maintained in sand, in an incubator under controlled conditions. The larvae molted to the nymphal stage without feeding, and five nymphal stages (N1, N2, N3, N4 and N5) were observed in both generations. In both generations emergence of adults started from N3 when sex ratio was 0.85:1 (23 males and 27 females) in F1 and 0.63:1 (34 males and 54 females) in F2. For both generations, N4 generated more females, while N5 only produced females. The pre-ecdysis period of the nymphs ranged from 31.1 to 38.6 days. Two gonotrophic cycles were observed, and the first one presented a higher average number of eggs deposited (N=139) than the second (N=73.8). The mean duration of the life cycle (egg to egg) of O. brasiliensis was 215.4 days for the first generation and 195 days for the second.

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In order to describe the developmental biology of the tick Argas neghmei Kohls & Hoogstraal under laboratory conditions, 40 females and 40 males were collected from chicken coops located in Calama (II Region, Chile). They were fed on chickens and maintained under two laboratory conditions: one group at 30 ± 5 ºC and 35 ± 5 percent RH and another at 27 ± 5 ºC and 80 ± 5 percent RH, both at 12: 12 h L:D photoperiod. The ticks were observed daily to determine larval feeding periods, preoviposition, oviposition, egg incubation as well as the frequency of egg laying, number of eggs laid, and percentage of larval hatching. Females did not lay eggs at 80 ± 5 percent RH, and data on the biology of this tick was obtained only at 35 ± 5 percent RH. The life cycle of A. neghmei lasted an average of 269 days. Feeding period of each nymphal stage as well as of adult females between oviposition events lasted less than a day. Females laid on average 1.8 egg batches and egg-laying period lasted on average 14 days, during which about 96 eggs were laid per female.

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