New records of soft ticks (Acari: Argasidae) from caves in Brazil, with a morphological study of Ornithodoros fonsecai and an analysis of the taxonomic status of Antricola inexpectata.

In this study, we report soft ticks from bat-inhabiting caves in different areas of Brazil. From 2010 to 2019, we collected 807 tick specimens from nine caves located in four Brazilian states among two biomes. Ticks were morphologically identified as Antricola guglielmonei (282 specimens), Ornithodoros cavernicolous (260 specimens), and Ornithodoros fonsecai (265 specimens). Whereas A. guglielmonei was collected on bat guano in hot caves, O. cavernicolous and O. fonsecai were collected in cracks and crevices on the walls of cold caves, sometimes in the same chamber. Morphological identifications were corroborated by molecular and phylogenetic analyses inferred from tick mitochondrial 16S rRNA gene partial sequences. The sequences of A. guglielmonei, O. cavernicolous and O. fonsecai collected in this study clustered with conspecific GenBank sequences from different localities of Brazil. Remarkably, a clade containing 12 sequences of O. fonsecai was clearly bifurcated, denoting a degree of genetic divergence (up to 5 %) of specimens from Cerrado/Atlantic Forest biomes with the specimens from the Caatinga biome. To further evaluate this divergence, we performed morphometric analysis of the larval stage of different O. fonsencai populations by principal component analysis, which indicated that the larvae from Caatinga populations were generally smaller than the larvae from other biomes. Some of the present A. guglielmonei specimens were collected from the type locality of Antricola inexpectata. Comparisons of these specimens with the type specimens of A. inexpectata and A. guglielmonei indicated that they could not be separated by their external morphology. Hence, we are relegating A. inexpectata to a synonym of A. guglielmonei. This proposal is corroborated by our phylogenetic analysis.

Apparent tick paralysis by Otobius megnini in a cat.

In this study, we report a rare case of tick paralysis in a cat induced by Otobius megnini infestation. An 11-month-old female cat was admitted to a private veterinary clinic in Luling Texas, USA presenting with depression, tachycardia, and flaccid paralysis of the entire body. The four recovered ticks were morphologically and molecularly identified as O. megnini nymphs. Following initial tick removal and treatment with 0.1% milbemycin oxime in the ear canal on the first day of hospitalization, and additional tick removal and topical selamectin treatment on the second day of hospitalization the animal gradually improved. The recovery of the cat after tick removal supports the diagnosis of tick toxicosis. While tick antiserum is not available in North America, prevention of tick infestation and tick-induced paralysis can be effectively accomplished using repellent collars and the compliant use of other ectoparasiticide products year-round.

Experimental transmission of a novel relapsing fever group Borrelia harbored by Ornithodoros octodontus (Ixodida: Argasidae) in Chile.

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.

Aural hematoma in lambs associated with Otobius megnini (Ixodida: Argasidae) infestation.

In this report we described a case of aural hematomas in three lambs associated with Otobius megnini (Ixodida: Argasidae) infestation. From April to May 2021, five 3-month-old Hampshire cross lambs presented with unilateral aural hematomas. Upon otoscopic examination, engorged soft ticks (O. megnini) were observed in the external ear canals of three of the five lambs. The remaining two lambs had lesions consistent with infestation and were in a shared environment and deemed likely to have been infected. The treatment of all animals was based on the drainage of the serosanguinous fluid through an incision in the internal space of the ear pinna. Upon physical inspection of the entire flock (n = 310), O. megnini infestation was observed in one additional animal that did not have a hematoma. Following animal and environmental ectoparasiticide treatment with permethrin, no recurrences or additional cases of aural hematomas were observed in the flock in the following two-year period. To the authors' knowledge, this is the first description of aural hematomas in lambs associated with O. megnini infestation with successful recovery after surgery and off-label acaricide treatment.

Bat ectoparasites: chigger mites (Trombiculidae), ticks (Ixodidae and Argasidae), and bugs (Cimicidae) in the Eastern Palaearctic.

Nine species-level taxa of bat ectoparasites, three chigger mites (Trombiculidae), three hard (Ixodidae), and one soft tick (Argasidae) species, as well as two bug (Cimicidae) species from nine bat species hosts were detected in the Eastern Palaearctic. Trombiculid larvae of Leptotrombidium schlugerae, Leptotrombidium album, and Ascoschoengastia latyshevi were first recorded on bats in the temperate zone of eastern Russia. L. schlugerae was more abundant than A. latyshevi in the same study sites in Eastern Siberia, and the main hosts of both chigger species were Plecotus ognevi and Eptesicus nilssonii. Ixodid ticks Dermacentor marginatus, Ixodes simplex, and Ixodes sp. were sampled from bats in Kazakhstan, the Far East, and Eastern Siberia, respectively. Phylogenetic analysis based on Cox1, 16S rDNA, and ITS2 sequences of I. simplex showed that the specimens from the Far East grouped into a clade distributed in the Eastern Palaearctic and India. In turn, the specimen of Ixodes sp. from Eastern Siberia was most closely related to Ixodes soricis and Ixodes angustus with p-distance of 9.8-10.7% (Cox1), suggesting that this tick probably belongs to a new species. Argas vespertilionis larvae were collected from three widespread bat species in Kazakhstan. Two bug species, Cimex pipistrelli and Cimex aff. lectularius, were recorded in the Far East and Eastern Siberia, respectively. Specimens from Transbaikalia were morphologically identified as Cimex lectularius. However, they differed from the latter by 12.5-12.9% of Cox1 sequences, indicating that C. aff. lectularius may be a new species.

Molecular detection of Borrelia sp. in Ornithodoros cavernicolous (Acari: Argasidae) in midwestern Brazil.

Ticks are obligate hematophagous parasites that can transmit to vertebrate hosts several pathogens, including viruses, bacteria, protozoa and helminths. Among these agents, some Borrelia species some Borrelia species cause disease in humans and other vertebrate hosts; therefore, they have medical and veterinary health importance. To gather additional information on Borrelia species in Brazil, the current study aimed to detect the presence of these species in Ornithodoros cavernicolous ticks collected in September 2019 from cement pipes that are used by bats as shelter in a farm located in the midwestern region of Brazil. DNA samples obtained from 18 specimens of O. cavernicolous were subjected of two polymerase chain reactions, targeting a segment of the Borrelia fla B gene. Of the samples tested, only one (6 %, 1/18) showed amplification. The nucleotide sequence of the amplified DNA showed more than 97 % (293/300) identity with a sequence of a Borrelia sp. detected in blood collected from a bat from Macaregua Cave, Colombia, and more than 97 % (292/300) detected in lungs from vampire bats from northeastern Brazil. The deduced amino acid sequences were identical to each other. Phylogenetic analysis indicated that these sequences formed a group of Borrelia species (putatively associated with bats) that is closely related to sequences of Borrelia species of the Lyme borreliosis group. Further investigations should be carried out in order to determine whether the sequence of the Borrelia sp. we found belongs to a new taxon. It will also be of great importance to determine which vertebrate hosts, besides bats, O. cavernicolous ticks can parasitize in order to investigate whether the Borrelia sp. we found may be transmitted and cause disease to the other vertebrate hosts.

Anti-tick vaccine candidate subolesin is important for blood feeding and innate immune gene expression in soft ticks.

Subolesin is a conserved molecule in both hard and soft ticks and is considered as an effective candidate molecule for the development of anti-tick vaccine. Previous studies have reported the role of subolesin in blood feeding, reproduction, development, and gene expression in hard ticks. However, studies addressing the role of subolesin in soft ticks are limited. In this study, we report that subolesin is not only important in soft tick Ornithodoros turicata americanus blood feeding but also in the regulation of innate immune gene expression in these ticks. We identified and characterized several putative innate immune genes including Toll, Lysozyme precursor (Lp), fibrinogen-domain containing protein (FDP), cystatin and ML-domain containing protein (MLD) in O. turicata americanus ticks. Quantitative real-time polymerase chain reaction analysis revealed the expression of these genes in both O. turicata americanus salivary glands and midgut and in all developmental stages of these soft ticks. Significantly increased expression of fdp was noted in salivary glands and midgut upon O. turicata americanus blood feeding. Furthermore, RNAi-mediated knockdown of O. turicata americanus subolesin expression affected blood feeding and innate immune gene expression in these ticks. Significant downregulation of toll, lp, fdp, cystatin, and mld transcripts was evident in sub-dsRNA-treated ticks when compared to the levels noted in mock-dsRNA-treated control. Collectively, our study not only reports identification and characterization of various innate immune genes in O. turicata americanus ticks but also provides evidence on the role of subolesin in blood feeding and innate immune gene expression in these medically important ticks.

Controlling Argas arboreus and A. persicus (Acari: Argasidae) by Adiantum capillus-veneris L. extracts with phytochemical analysis.

Argas species are parasites associated mostly with birds. Their infestations of the host may cause blood loss, resulting in anemia and finally death. Egypt loses millions of tons annually from birds because of these parasites. In addition, they can transmit pathogens to animals and humans. The acaricidal effects of the ethanolic and methanolic extracts of Adiantum capillus-veneris at different concentrations (1-4%) against semi-fed adults of Argas arboreus and A. persicus were investigated during 30 days after treatments. Mobility and mortality, acaricide efficacy, and the concentration that kills 50% of specimens (LC50) were estimated. The percentage of dead adults of both Argas species appeared during 6 days considerably until 30 days was significantly increased after treatment of either ethanol or methanol extracts of Adiantum at 1-4%, versus control groups. Ethanolic extracts (100% mortality) were more effective than methanolic ones (90% mortality) for both Argas species. Argas arboreus (80% efficacy and 5.9% LC50) was more resistant than A. persicus (100% efficacy and 4.1% LC50). Generally, males were more resistant than females. The chemical profile (by gas chromatography-mass spectrometry analysis) for the ethanolic extract of Ad. capillus-veneris at 4% (the most effective extract) was provided for the first time, which showed that the major group was sugars and sugar alcohols, and the main components were thymol-ß-d-glucopyranoside, D-(-)-Tagatofuranose, D-Arabinose, D-Galactose, D-(-)-Fructofuranose and Anthracene, 1-methyl. The efficiency of all these components was discussed. Based on the findings, bioactive compounds present in Ad. capillus-veneris have the potential to be applied as substitutes for synthetic acaricides and a biological control agent in the management of A. arboreus and A. persicus ticks.

Autogenous reproduction by Ornithodoros turicata (Ixodida: Argasidae) females and vertical transmission of the tick-borne pathogen Borrelia turicatae (Spirochaetales: Borreliaceae).

IMPORTANCE: Previous research has implicated Ornithodoros ticks, including Ornithodoros turicata, as long-term reservoirs of relapsing fever (RF) spirochetes. Considering the tick's long lifespan and their efficiency in maintaining and transferring spirochetes within the population, the infection could persist in a given enzootic focus for decades. However, little is known about the relative importance of horizontal and vertical transmission routes in the persistence and evolution of RF Borrelia. Our observations on the reproductive biology of O. turicata in the absence of vertebrate hosts indicate an additional mechanism by which Borrelia turicatae can be maintained in the environment. This work establishes the foundation for studying O. turicata reproduction and spirochete-vector interactions, which will aid in devising control measures for Ornithodoros ticks and RF spirochetes.

New records of Ornithodoros turicata (Ixodida: Argasidae) in rural and urban sites in the Mexican states of Aguascalientes and Zacatecas indicate the potential for tick-borne relapsing fever.

Soft ticks from the Ornithodoros genus are vectors of relapsing fever (RF) spirochetes around the world. In Mexico, they were originally described in the 19th century. However, few recent surveillance studies have been conducted in Mexico, and regions where RF spirochetes circulate remain vague. Here, the presence of soft ticks in populated areas was assessed in two sites from the Mexican states of Aguascalientes and Zacatecas. Argasidae ticks were collected, identified by morphology and mitochondrial 16S rDNA gene sequencing, and tested for RF borreliae. The specimens in both sites were identified as Ornithodoros turicata but no RF spirochetes were detected. These findings emphasize the need to update the distribution of these ticks in multiple regions of Mexico and to determine the circulation of RF borreliosis in humans and domestic animals.

Ticks (Ixodida) associated with bats (Chiroptera): an updated list with new records for Brazil.

Bats harbor diverse groups of ectoparasites, such as insects and mites like ticks (Ixodida). Some species of ticks with records for bats and humans have already been reported with the occurrence of pathogens. This research article aims to document new geographical and host records of ticks infesting bats in Rio de Janeiro state, Southeastern Brazil, and provides a list of tick species associated with bats in Brazil. We counted 12 argasid ticks and five ixodid ticks associated with six individuals of bats. Larvae of Amblyomma sp., Ixodes sp., Ornithodoros sp., and Ornithodoros hasei and one nymph of Amblyomma sculptum parasitizing Artibeus obscurus, Phyllostomus hastatus, Micronycteris sp., Molossus fluminensis, and Carollia perspicillata in different localities of Rio de Janeiro state were studied. We carried out a systematic review with the descriptors: tick bat Brazil. We considered data from 42 articles in the systematic review. We compiled eleven records of Ixodidae, and 160 records of Argasidae. Ornithodoros cavernicolous were the most recorded tick species. Overall, we registered 171 tick-bat or roost-bat associations with 85 records of these infesting bats. The review also shows the occurrence of tick species associated with bats, and we present new records on ticks parasitizing bats in Brazil.

Description of a new Pavlovskyella species (Acari: Argasidae) from Chile.

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.

Distribution of the soft tick Carios vespertilionis in lowlands and low mountain regions of Germany.

In Germany, the knowledge about ticks infesting bats is limited, and is restricted only to a few studies, most of them dating back decades. To further improve our knowledge on ticks parasitising bats, healthy and sick bats in central Germany were examined for ticks. In total 519 larvae and one nymph of Carios vespertilionis were collected from nine bat species: Eptesicus nilssonii, Eptesicus serotinus, Myotis daubentonii, Myotis myotis, Nyctalus leisleri, Pipistrellus nathusii, Pipistrellus pygmaeus, Pipistrellus pipistrellus, and Vespertilio murinus. Either the presence of C. vespertilionis was new for some areas or it was confirmed in some federal states in central Germany. The infestation rate was mostly low (n = 1-5 larvae/bat). However, in two cases a high number of ticks was observed. The highest infestation of 97 C. vespertilionis larvae was recorded on one Parti-coloured bat (V. murinus).

Soft Tick Relapsing Fever - United States, 2012-2021.

Soft tick relapsing fever (STRF) (also known as tickborne relapsing fever) is a rare infection caused by certain Borrelia spirochetes and transmitted to humans by soft-bodied Ornithodoros ticks. In the United States, acquisition of STRF is commonly associated with exposure to rustic cabins, camping, and caves. Antibiotic treatment is highly effective for STRF, but without timely treatment, STRF can result in severe complications, including death. No nationally standardized case definition for STRF exists; however, the disease is reportable in 12 states. This report summarizes demographic and clinical information for STRF cases reported during 2012-2021 from states where STRF is reportable. During this period, 251 cases were identified in 11 states. The median annual case count was 24. Most patients with STRF (55%) were hospitalized; no fatalities were reported. The geographic distribution and seasonal pattern of STRF have remained relatively constant since the 1990s. Persons should avoid rodent-infested structures and rodent habitats, such as caves, in areas where STRF is endemic. STRF surveillance, prevention, and control efforts would benefit from a standardized case definition and increased awareness of the disease among the public and clinicians.

The Soft Tick Ornithodoros moubata and Its Role in the Epidemiology of African Swine Fever in Central Malawi.

African swine fever is a viral hemorrhagic disease of pigs (Sus scrofa) caused by a double-stranded DNA virus, African swine fever virus (ASFV). With up to 100% mortality in pigs and no vaccine, the socioeconomic impacts of this disease are immense. In sub-Saharan Africa, warthogs (Phacochoerus africanus) are the original vertebrate host for the virus, which is transmitted among wild suids via the soft tick Ornithodoros moubata. In Malawi, O. moubata is thought to be widely distributed, with the potential to spread ASFV beyond its historical enzootic zone in the Central Region. We surveyed 76 domestic pig farms, 18 active warthog burrows, and three bushpigs (Potamochoerus larvatus) resting sites for O. moubata in the Central Region of Malawi, and tested the ticks for ASFV using PCR. We found a limited distribution for O. moubata: 75 ticks were discovered at a single farm in the Mchinji District. Eleven percent of these ticks were ASFV positive. This suggests that wildlife and O. moubata play a limited role in the epidemiology of ASF in Malawi; thus, other factors for disease spread, such as fomites and pig-to-pig infection, need to be considered.

Argas ticks (Ixodida: Argasidae) on migratory birds from Africa: first record of a genotype close to Argas africolumbae in Italy.

In spring, migratory birds reach Europe, mainly from sub-Saharan Africa or from northern African countries. Avian species may be implicated in the spread of pathogens, either as reservoirs, hosts or carriers of infected ectoparasites. In 2021, on Ventotene Island (Latium region, Italy) within a project focused on the potential incoming pathogens via migratory birds from Africa, we found two larvae of Argas sp., on the redstart Phoenicurus phoenicurus, that shared morphological features with the African Argas (Argas) africolumbae. Comparison of the tested larval DNA sequences to the adult reference sequences showed the highest identity (> 92%) with homologous sequences of A. africolumbae collected in South Africa and in Spain. This study reports the first detection of Argas africolumbae-like specimens in Italy.

Reticulinasus salahi (Acarina: Argasidae), a tick of bats and man in the Palaearctic and Afrotropics: review of records with the first pathogens detected.

The soft ticks of the genus Reticulinasus Schulze, 1941 (family Argasidae Koch, 1844) are ectoparasites of the fruit bats of the Old World (Pteropodidae). Reticulinasus salahi (Hoogstraal, 1953) is the only representative of this genus that occurs in the western part of the Palaearctic. This unusual distribution reflects the distributon range of its primary host, Rousettus aegyptiacus (Geoffroy, 1810). In this contribution, we present a revised review of records of this tick that were made in two periods, 1951-1966 (records from Egypt, Israel, Jordan, Spain) and 2005-2019 (Cyprus, Iran, Oman), and additionally, we present notes, re-determinations, new records, and summary of hosts of this tick. Besides the primary host, the revised list of hosts comprises two bats (Taphozous perforatus Geoffroy, 1818, Otonycteris hemprichii Peters, 1859) and the human (Homo sapiens Linnaeus, 1758). We also tried to identify pathogens in specimens of this tick collected from R. aegyptiacus in Oman. The DNA of the Mouse herpesvirus strain 68 (MHV-68), of two bacteria, Borellia burgdorferii sensu lato, and Ehrlichia sp. almost identical (98%) with Candidatus Ehrlichia shimanensis was detected in several larvae specimens.

Ticks of Australasia: 125 species of ticks in and around Australia.

The present work is an update and a substantial expansion of Barker & Walker (2014). Barker & Walker (2014) dealt with 16 species of particular importance to domestic animals and humans whereas the present work deals with all 125 of the species known from Australasia; that is Australia, New Zealand, the island of New Guinea, Sulawesi, and the islands of the western Pacific. These comprise 102 species of ixodid (hard) ticks and 23 species of argasid (soft) ticks.

Atlas of ticks (Acari: Argasidae, Ixodidae) in Germany: 1st data update.

The first data update of the atlas of ticks in Germany published in 2021 is presented here. This atlas provides maps based on georeferenced tick locations of 21 species endemic in Germany as well as three tick species that are regularly imported to Germany. The data update includes the following numbers of newly georeferenced tick locations: 17 Argas reflexus, 79 Carios vespertilionis, 2 Dermacentor marginatus, 43 Dermacentor reticulatus, 4 Haemaphysalis concinna, 3 Haemaphysalis punctata, 3 Hyalomma rufipes, 3 Ixodes apronophorus, 9 Ixodes arboricola, 1 Ixodes ariadnae, 30 Ixodes canisuga, 3 Ixodes frontalis, 80 Ixodes hexagonus, 3 Ixodes lividus, 497 Ixodes ricinus/inopinatus, 1 Ixodes rugicollis, 17 Ixodes trianguliceps, 14 Ixodes vespertilionis, and 45 Rhipicephalus sanguineus sensu lato. Old and new tick findings were mapped, such as the northernmost occurrence of D. marginatus in Germany observed in 2021, but also the historical records from the first descriptions of I. apronophorus and I. arboricola, which were georeferenced here for the first time. The digital dataset of tick locations available for Germany is supplemented by 854 new tick locations. These records increase the number of tick species mapped in the federal states Bavaria, Brandenburg and Mecklenburg Western Pomerania by five each, those in Berlin and Schleswig-Holstein by four each, those in Hamburg by three, those in Baden-Wuerttemberg, Bremen, Lower Saxony, Northrhine-Westphalia, Rhineland Palatinate and Thuringia by two each, and those in Hesse, Saxony and Saxony-Anhalt by one each. Thus, the first data update of the tick atlas in Germany and the underlying digital dataset significantly improve our knowledge of the distribution of these tick species and helps to investigate the effects of climate change and habitat changes on them.

A serological assay to detect and differentiate rodent exposure to soft tick and hard tick relapsing fever infections in the United States.

Human cases of relapsing fever (RF) in North America are caused primarily by Borrelia hermsii and Borrelia turicatae, which are spread by argasid (soft) ticks, and by Borrelia miyamotoi, which is transmitted by ixodid (hard) ticks. In some regions of the United States, the ranges of the hard and soft tick RF species are known to overlap; in many areas, recorded ranges of RF spirochetes overlap with Lyme disease (LD) group Borrelia spirochetes. Identification of RF clusters or cases detected in unusual geographic localities might prompt public health agencies to investigate environmental exposures, enabling prevention of additional cases through locally targeted mitigation. However, exposure risks and mitigation strategies differ among hard and soft tick RF, prompting a need for additional diagnostic strategies that differentiate hard tick from soft tick RF. We evaluated the ability of new and previously described recombinant antigens in serological assays to differentiate among prior exposures in mice to LD, soft or hard tick RF spirochetes. We extracted whole-cell protein lysates from RF Borrelia cultures and synthesized six recombinant RF antigens (Borrelia immunogenic protein A (BipA) derived from four species of RF Borrelia, glycerophosphodiester phosphodiesterase (GlpQ), and Borrelia miyamotoi membrane antigen A (BmaA)) to detect reactivity in laboratory derived (Peromyscus sp. and Mus sp.) mouse serum infected with RF and LD Borrelia species. Among 44 Borrelia exposed mouse samples tested, all five mice exposed to LD spirochetes were correctly differentiated from the 39 mice exposed to RF Borrelia using the recombinant targets. Of the 39 mice exposed to RF spirochetes, 28 were accurately categorized to species of exposure (71%). Segregation among soft tick RF species (Borrelia hermsii, Borrelia parkeri and Borrelia turicatae) was inadequate (58%) owing to observed cross-reactivity among recombinant BipA protein targets. However, among the 28 samples accurately separated to species, all were accurately assigned to soft tick or hard tick RF type. Although not adequately specific to accurately categorize exposure to soft tick RF species, the recombinant BipA protein targets from soft and hard tick RF species show utility in accurately discriminating mouse exposures to LD or RF Borrelia, and accurately segregate hard tick from soft tick RF Borrelia exposure.