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Background: Bacteria of the genus Borrelia are agents of disease in both domestic animals and humans and pose a significant public health risk. Borrelia species have complex transmission cycles, often using rodents as vertebrate reservoir hosts. These bacteria are classified into three well-defined monophyletic groups: Borrelia burgdorferi sensu lato (Bbsl) complex, the relapsing fever (RF) group, and a third group associated with reptiles and echidnas. Moreover, a new group of Borrelia associated with rodents has recently been proposed, as these bacteria form a phylogenetic group separated from the previously mentioned groups. This study aimed to investigate the presence of DNA of Borrelia spirochetes in rodents in specific areas of the Colombian Caribbean., Methods: A total of 155 rodent spleen samples were selected from the tissue bank. These samples were obtained in the departments of La Guajira and Córdoba (Northern Colombia). DNA extraction and specific real-time polymerase chain reaction (PCR) targeting Borrelia 16S ribosomal RNA (rRNA) gene were performed, followed by nested PCR (nPCR) on positive samples to obtain larger fragments of the 16S rRNA gene and characterize the flaB gene. Alignments of generated sequences and ortholog sequences downloaded from Genbank were performed in Clustal Omega. A phylogenetic tree was built with the maximum likelihood method in IQTREE., Results: Spleen samples from rodents of the genera Heteromys, Mus, Necromys, Olygoryzomys, Proechymis, Rattus, Sigmodon, and Zygodontomys were processed. Overall, 6.5% (4/162) of the animals tested positive for Borrelia by real-time PCR. All quantitative PCR (qPCR)-positive samples were also positive for nPCR targeting the 16S rRNA gene, yielding fragments of 344-408 bp and 603-673 bp from two Sigmodon rodents and two Zygodontomys rodents from La Guajira and Córdoba. All samples were negative for the flaB gene. Only samples from Zygodontomys rodents presented good quality sequences. A BLASTn analysis showed a percentage of identity ranging between 98.16 and 96.06% with Borrelia sp. R57. Phylogenetic analysis revealed that sequences of the present study clustered with species of the recently proposed Borrelia "rodent group.", Conclusions: This is the first detection of borreliae of the "rodent group" in South America. Our results reaffirm the occurrence of a group of spirochetes associated with rodents, extending its geographic distribution to the Colombian Caribbean., Competing Interests: Declarations. Ethics approval and consent to participate: The rodents were captured under the permits of the National Authority of Environmental Licenses [ANLA], resolution no. 00914. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests., (© 2024. The Author(s).)

The impact of tick-borne diseases caused by pathogens such as Anaplasma phagocytophilum, Neoehrlichia mikurensis, Borrelia miyamotoi, Rickettsia helvetica and Babesia species on public health is largely unknown. Data on the prevalence of these pathogens in Ixodes ricinus ticks from seven countries within the North Sea Region in Europe as well as the types and availability of diagnostic tests and the main clinical features of their corresponding diseases is reported and discussed. Raised awareness is needed to discover cases of these under-recognized types of tick-borne disease, which should provide valuable insights into these diseases and their clinical significance., Competing Interests: Declaration of Competing Interest None, (Copyright © 2023. Published by Elsevier GmbH.)

Background: Small mammals play an important role in the life-cycle of ticks and are reservoirs for several zoonotic pathogens. The aim of this study was to provide epidemiological data regarding the presence of Borrelia spp. in tissues of small mammals from Romania., Methods: We examined 401 individuals belonging to 11 small mammal species collected in Romania. Collections cover the largest effort to survey these reservoirs in the country. Tissue samples were analyzed by multiplex qPCR targeting the ospA gene of Borrelia burgdorferi (s.l.) and a part of the flaB gene of B. miyamotoi. Positive samples were further analysed by conventional PCR and sequenced., Results: The overall prevalence of infection with Borrelia spp. in small mammal tissues was 4.9%. The most commonly detected species were B. afzelii, followed by B. garinii/B. bavariensis, B. miyamotoi and B. burgdorferi (s.s.). To our knowledge, we report for the first time the detection of Borrelia spp. in Crocidura leucodon and C. suaveolens, and B. miyamotoi in the liver of Myodes glareolus., Conclusions: To our knowledge, our study evaluates for the first time the occurrence of Borrelia spp. in small mammals in Romania, contributing to a better knowledge of the distribution of these bacteria. This survey upgrades previous data on the spatial distribution of the pathogens and reveals the importance of animal surveillance regarding Lyme borreliosis and relapsing fever caused by B. miyamotoi.

Surveillance to investigate the wildlife-vector transmission cycle of the human pathogen Borrelia miyamotoi in California, USA, revealed infections in dusky-footed woodrats, brush mice, and California mice. Phylogenetic analyses suggest a single, well-supported clade of B. miyamotoi is circulating in California.

Background: Elucidating which wildlife species significantly contribute to the maintenance of Ixodes ricinus populations and the enzootic cycles of the pathogens they transmit is imperative in understanding the driving forces behind the emergence of tick-borne diseases. Here, we aimed to quantify the relative contribution of four mustelid species in the life-cycles of I. ricinus and Borrelia burgdorferi (sensu lato) in forested areas and to investigate their role in the transmission of other tick-borne pathogens. Road-killed badgers, pine martens, stone martens and polecats were collected in Belgium and the Netherlands. Their organs and feeding ticks were tested for the presence of tick-borne pathogens., Results: Ixodes hexagonus and I. ricinus were found on half of the screened animals (n = 637). Pine martens had the highest I. ricinus burden, whereas polecats had the highest I. hexagonus burden. We detected DNA from B. burgdorferi (s.l.) and Anaplasma phagocytophilum in organs of all four mustelid species (n = 789), and Neoehrlichia mikurensis DNA was detected in all species, except badgers. DNA from B. miyamotoi was not detected in any of the investigated mustelids. From the 15 larvae of I. ricinus feeding on pine martens (n = 44), only one was positive for B. miyamotoi DNA, and all tested negative for B. burgdorferi (s.l.), N. mikurensis and A. phagocytophilum. The two feeding larvae from the investigated polecats (n = 364) and stone martens (n = 39) were negative for all four pathogens. The infection rate of N. mikurensis was higher in feeding nymphs collected from mustelids compared to questing nymphs, but not for B. burgdorferi (s.l.), B. miyamotoi or A. phagocytophilum., Conclusions: Although all stages of I. ricinus can be found on badgers, polecats, pine and stone martens, their relative contribution to the life-cycle of I. ricinus in forested areas is less than 1%. Consequently, the relative contribution of mustelids to the enzootic cycles of I. ricinus-borne pathogens is negligible, despite the presence of these pathogens in organs and feeding ticks. Interestingly, all four mustelid species carried all stages of I. hexagonus, potentially maintaining enzootic cycles of this tick species apart from the cycle involving hedgehogs as main host species.

To understand the epidemiology of tick infestation and tick-borne diseases in pet dogs in south-eastern China and to develop a reference for their prevention and treatment, we collected 1550 ticks parasitizing 562 dogs in 122 veterinary clinics from 20 cities of south-eastern China. Dogs were tested for common tick-borne pathogens; collected ticks were identified and processed for the detection of tick-borne pathogens. The use of an in vitro ELISA diagnostic kit for antibody detection (SNAP®4Dx® Plus) on dog sera found the infection rates with Borrelia burgdorferi sensu lato, Ehrlichia canis, and Anaplasma spp. to be 0.4%, 1.3% and 2.7%, respectively. By using a specific ELISA method, the infection rate with Babesia gibsoni was 3.9%. Rhipicephalus sanguineus sensu lato, Haemaphysalis longicornis and Rhipicephalus haemaphysaloides were the major tick species identified on pet dogs. PCR tests were conducted to detect five tick-borne pathogens in 617 ticks. The infection rate was 10.2% for E. canis, 3.4% for Anaplasma platys, 2.3% for B. gibsoni, 0.3% for B. burgdorferi s.l. and 0% for Babesia canis. Some ticks were co-infected with two (1.46%) or three pathogens (0.16%). These results indicate the infestation of pet dogs by ticks infected with tick-borne pathogens in south-eastern China, and the need for effective treatment and routine prevention of tick infestations in dogs., (© J. Zhang et al., published by EDP Sciences, 2017.)

Background: In the United States, Tick-Borne Relapsing Fever (TBRF) in dogs is caused by the spirochete bacteria Borrelia turicatae and Borrelia hermsii, transmitted by Ornithodoros spp. ticks. The hallmark diagnostic feature of this infection is the visualization of numerous spirochetes during standard blood smear examination. Although the course of spirochetemia has not been fully characterized in dogs, in humans infected with TBRF the episodes of spirochetemia and fever are intermittent., Objectives: To describe TBRF in dogs by providing additional case reports and reviewing the disease in veterinary and human medicine., Animals: Five cases of privately-owned dogs naturally infected with TBRF in Texas are reviewed., Methods: Case series and literature review., Results: All dogs were examined because of lethargy, inappetence, and pyrexia. Two dogs also had signs of neurologic disease. All dogs had thrombocytopenia and spirochetemia. All cases were administered tetracyclines orally. Platelet numbers improved and spirochetemia and pyrexia resolved in 4 out of 5 dogs, where follow-up information was available., Conclusion and Clinical Importance: TBRF is likely underdiagnosed in veterinary medicine. In areas endemic to Ornithodoros spp. ticks, TBRF should be considered in dogs with thrombocytopenia. Examination of standard blood smears can provide a rapid and specific diagnosis of TBRF when spirochetes are observed., (Copyright © 2016 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.)

North Korea is located on the northern part of the Korean Peninsula in East Asia. While tick-borne pathogens of medical and veterinary importance have been reported from China and South Korea, they have not been reported from North Korea. To screen for zoonotic tick-borne pathogens in North Korea, ticks were collected from domestic goats. A total of 292 (27 nymph, 26 male, 239 female) Haemaphysalis (H.) longicornis were collected and assayed individually for selected tick-borne pathogens. A total of 77 (26.4%) were positive for Anaplasma bovis, followed by Bartonella (B.) grahamii (15, 5.1%), Anaplasma phagocytophilum (12, 4.1%), Bartonella henselae (10, 3.4%), and Borrelia spp. (3, 1.0%) based on 16S ribosomal RNA and ITS species-specific nested polymerase chain reaction. Using the groEL-based nested PCR, a total of 6 and 1 H. longicornis were positive for B. grahamii and B. henselae, respectively. All products were sequenced and demonstrated 100% identity and homology with previously reported sequences from other countries in GenBank. This is the first report of the detection of tick-borne pathogens in the North Korea and suggests that farm animals may act as reservoirs for zoonotic tick-borne pathogens.

A serological survey of Borrelia infection of dogs was performed in Sapporo, Japan, where Borrelia garinii infection in dogs was detected in 2011. A total of 314 serum samples were collected from dogs that visited three animal hospitals in Sapporo from 2012 to 2014. The two-step evaluation method, involving screening ELISA followed by Western blot analysis, was used to detect antibodies against Borrelia species. A total of 34 samples were positive by ELISA. Among those 34 samples, 32 were positive for Borrelia spp. by Western blot. These findings suggest that the 32 dogs (10.2%) generated antibodies against Borrelia burgdorferi sensu lato, such as B. garinii or B. afzelii. Antibody positivity was 7.6% and 13.3% for dogs living in urban and rural areas, respectively. Dogs with a history of tick infestation showed a positive rate of 16.7%, which was higher, although not significantly, than the 6.7% among dogs without a history.

Classifying the states of an individual and quantifying transitions between states are crucial while modeling animal behavior, movement, and physiologic status. When these states are hidden or imperfectly known, it is particularly convenient to relate them to appropriate quantitative measurements taken on the individual. This task is, however, challenging when quantitative measurements are not available at each sampling occasion. For capture-recapture data, various ways of incorporating such non-discrete information have been used, but they are either ad hoc and/or use a fraction of the available information by relying on a priori thresholds to assign individual states. Here we propose assigning discrete states based on a continuous measurement, and then modeled survival and transition probabilities based on these assignments. The main advantage of this new approach is that a more informative use of the non-discrete information is done. As an illustrative working example, we applied this approach to eco-epidemiological data collected across a series of years in which individuals of a long-lived seabird, the Black-legged Kittiwake (Rissa tridactyla), could either be visually detected or physically recaptured and blood sampled for subsequent immunological analyses. We discuss how this approach opens many perspectives in eco-epidemiology, but also more broadly, in population ecology.

A total of 87 Thoroughbred horses and 10 ixodid ticks from a ranch in Hidaka district, Hokkaido were tested for tick-borne diseases. Using the indirect fluorescent antibody (IFA) method, 3.4, 92.0 and 97.7% of the horses showed antibody titers of ≥ 80 against Anaplasma phagocytophilum, Rickettsia helvetica, and Borrelia garinii, respectively. This is the first report of infection with the 3 pathogens in horses in Japan. Using PCR, DNAs from the peripheral blood of all horses were found negative with any Anaplasma, Rickettsia and Borrelia spp., while those from Haemaphysalis megaspinosa ticks were found positive for Anaplasma sp. closely related to A. phagocytophilum in Japan, and A. bovis. B. japonica was also detected in an H. flava tick for the first time.

Background: In the Eastern and Upper Midwestern regions of North America, Ixodes scapularis (L.) is the most abundant tick species encountered by humans and the primary vector of B. burgdorferi, whereas in the southeastern region Amblyomma americanum (Say) is the most abundant tick species encountered by humans but cannot transmit B. burgdorferi. Surveys of Borreliae in ticks have been conducted in the southeastern United States and often these surveys identify B. lonestari as the primary Borrelia species, surveys have not included Arkansas ticks, canines, or white-tailed deer and B. lonestari is not considered pathogenic. The objective of this study was to identify Borrelia species within Arkansas by screening ticks (n=2123), canines (n=173), and white-tailed deer (n=228) to determine the identity and locations of Borreliae endemic to Arkansas using PCR amplification of the flagellin (flaB) gene., Methods: Field collected ticks from canines and from hunter-killed white-tailed were identified to species and life stage. After which, ticks and their hosts were screened for the presence of Borrelia using PCR to amplify the flaB gene. A subset of the positive samples was confirmed with bidirectional sequencing., Results: In total 53 (21.2%) white-tailed deer, ten (6%) canines, and 583 (27.5%) Ixodid ticks (252 Ixodes scapularis, 161 A. americanum, 88 Rhipicephalus sanguineus, 50 Amblyomma maculatum, 19 Dermacentor variabilis, and 13 unidentified Amblyomma species) produced a Borrelia flaB amplicon. Of the positive ticks, 324 (22.7%) were collected from canines (151 A. americanum, 78 R. sanguineus, 43 I. scapularis, 26 A. maculatum, 18 D. variabilis, and 8 Amblyomma species) and 259 (37.2%) were collected from white-tailed deer (209 I. scapularis, 24 A. maculatum, 10 A. americanum, 10 R. sanguineus, 1 D. variabilis, and 5 Amblyomma species). None of the larvae were PCR positive. A majority of the flaB amplicons were homologous with B. lonestari sequences: 281 of the 296 sequenced ticks, 3 canines, and 27 deer. Only 22 deer, 7 canines, and 15 tick flaB amplicons (12 I. scapularis, 2 A. maculatum, and 1 Amblyomma species) were homologous with B. burgdorferi sequences., Conclusions: Data from this study identified multiple Borreliae genotypes in Arkansas ticks, canines and deer including B. burgdorferi and B. lonestari; however, B. lonestari was significantly more prevalent in the tick population than B. burgdorferi. Results from this study suggest that the majority of tick-borne diseases in Arkansas are not B. burgdorferi.

Background: The presence in Morocco of Argasid ticks of the Ornithodoros erraticus complex, the vector of tick-borne relapsing fever (TBRF) in North Africa, has been known since 1919, but the disease is rarely diagnosed and few epidemiological data are available., Methodology/principal Findings: Between 2006 and 2011, we investigated the presence of Ornithodoros ticks in rodent burrows in 34 sites distributed across Morocco. We also collected small mammals in 10 sites and we investigated TBRF in febrile patients in Kenitra district. The prevalence of Borrelia infections was assessed by nested PCR amplification in ticks and the brain tissue of small mammals, and by evaluation of thick blood films in patients. A high proportion of burrows were infested with ticks of the O. erraticus complex in all regions of Morocco, with a mean of 39.5% for the whole country. Borrelia infections were found in 39/382 (10.2%) of the ticks and 12/140 (8.6%) of the rodents and insectivores studied by PCR amplification, and 102 patients tested positive by thick blood film. Five small mammalian species were found infected: Dipodillus campestris, Meriones shawi, Gerbillus hoogstrali, Gerbillus occiduus and Atelerix algirus. Three Borrelia species were identified in ticks and/or rodents: B. hispanica, B. crocidurae and B. merionesi., Conclusions/significance: Tick populations belonging to O. erraticus complex are widely distributed in Morocco and a high proportion of ticks and small mammals are infected by Borrelia species. Although rarely diagnosed, TBRF may be a common cause of morbidity in all regions of Morocco.

Thirty-five deer mice, Peromyscus maniculatus, were trapped on Gannet Cluster 2 (GC-2), one of a group of islands numbered by convention in the Gannet Island Archipelago, and examined for ectoparasites. One species each of Acari (Ixodes uriae) and Siphonaptera (Orchopeas leucopus) were recovered. Samples of mice favored males to females (3.4∶1). Twenty-nine percent (10) of the mice were free of ectoparasites. Males were more heavily parasitized than females when both parasites were considered. No ticks were recovered from the female mice, while the males that were parasitized carried adult Ixodes uriae. These 2 ectoparasites parasitizing P. maniculatus, which is a known reservoir host for Lyme disease (Borrelia burgdorferi), may carry B. garinii and their presence would have serious implications for the spread of this human pathogen northward in continental North America.

An increasing number of studies reveal that ticks and their hosts are infected with multiple pathogens, suggesting that coinfection might be frequent for both vectors and wild reservoir hosts. Whereas the examination of associations between coinfecting pathogen agents in natural host-vector-pathogen systems is a prerequisite for a better understanding of disease maintenance and transmission, the associations between pathogens within vectors or hosts are seldom explicitly examined. We examined the prevalence of pathogen agents and the patterns of associations between them under natural conditions, using a previously unexamined host-vector-pathogen system--green lizards Lacerta viridis, hard ticks Ixodes ricinus, and Borrelia, Anaplasma, and Rickettsia pathogens. We found that immature ticks infesting a temperate lizard species in Central Europe were infected with multiple pathogens. Considering I. ricinus nymphs and larvae, the prevalence of Anaplasma, Borrelia, and Rickettsia was 13.1% and 8.7%, 12.8% and 1.3%, and 4.5% and 2.7%, respectively. The patterns of pathogen prevalence and observed coinfection rates suggest that the risk of tick infection with one pathogen is not independent of other pathogens. Our results indicate that Anaplasma can play a role in suppressing the transmission of Borrelia to tick vectors. Overall, however, positive effects of Borrelia on Anaplasma seem to prevail as judged by higher-than-expected Borrelia-Anaplasma coinfection rates.

Lyme disease is a tick borne zoonotic infection, caused by Borrelia burgdorferi s.l. bacteria. For the transmission of the disease, the presence of ticks is a prerequisite. Lyme borreliosis mostly occurs in people and dogs, but it may occur in other animals. Ticks which carry B. burgdorferi s.l. in Serbia are of the Ixodes ricinus specis. In Serbia, Lyme disease was detected for the first time in the late '80-es. In dogs, clinical symptoms may occur even months after a tick bite, and include weakness, lymphadenopathy, fever, lameness, arthritis, etc. In our survey, we have observed tick and dog populations in the province of Vojvodina (northern part of Serbia). I. ricinus ticks were collected and examined for the presence of B. burgdorferi s.l. in several chosen locations. In addition, blood samples were collected from house dogs and pets from the same locations, and analyzed for the presence of antibodies specific for B. burgdorferi s.l. The results showed a mean infection of ticks of 22.12%, and a mean seroprevalence of Lyme disease in dogs of 25.81%. We conclude that in Vojvodina there is an actual risk of Lyme borreliosis for other animals and humans, because of the persistence of B. burgdorferi s.l. in both tick and dog populations.

The Lyme borreliosis agent Borrelia burgdorferi and the relapsing fever group species Borrelia miyamotoi co-occur in the United States. We used species-specific, quantitative polymerase chain reaction to study both species in the blood and skin of Peromyscus leucopus mice and host-seeking Ixodes scapularis nymphs at a Connecticut site. Bacteremias with B. burgdorferi or B. miyamotoi were most prevalent during periods of greatest activity for nymphs or larvae, respectively. Whereas B. burgdorferi was 30-fold more frequent than B. miyamotoi in skin biopsies and mice had higher densities of B. burgdorferi densities in the skin than in the blood, B. miyamotoi densities were higher in blood than skin. In a survey of host-seeking nymphs in 11 northern states, infection prevalences for B. burgdorferi and B. miyamotoi averaged approximately 0.20 and approximately 0.02, respectively. Co-infections of P. leucopus or I. scapularis with both B. burgdorferi and B. miyamotoi were neither more nor less common than random expectations.

This study reports the experimental transmission of Borrelia anserina to domestic chickens by infected Argas (Persicargas) miniatus. Clinical alterations as well as prepatent and patent periods were evaluated. Twenty-seven 67-day-old birds were divided into three groups in a randomized experimental design. The first group was exposed to ticks infected with B. anserina, the second group was exposed to noninfected ticks, and the third group was not exposed to ticks. Blood smears from each bird of groups 1 and 2 were prepared daily and examined for 25 days postexposure (PE). Examination of the blood smears from birds in group 1 revealed large numbers of spirochetes from days 5 to 12 PE. In this group the prepatent and patent periods were 5-7 and 4-7 days, respectively. Birds from group 1 presented ruffled feathers, pale combs, drowsiness, inappetence, loss of weight, and greenish diarrhea after day 6 PE. The current study confirms the viability of experimental transmission of B. anserina to domestic chickens by A. (P.) miniatus.

Borrelia burgdorferisensu lato infection has been confirmed in clinical cases in the northeast of Mexico; however, the bacterium has not been identified as infecting the tick vector Ixodes, Amblyomma and Dermacentor ticks were collected from mammals and plants in northeastern Mexico and examined for Borrelia. Eighteen of 214 ticks were PCR-positive for the fla and 16S rRNA genes and 15 for the ospA gene. Southern blotting with a fla probe and sequencing of ospA genes confirmed infection with B. burgdorferi sensu stricto. These findings, together with reports of indigenous cases, fulfil the criteria that allow northeastern Mexico to be considered as a zone endemic for Lyme disease.

The effect of introduced large herbivores on the abundance of Ixodes ricinus ticks and their Borrelia infections was studied in a natural woodland in The Netherlands. Oak and pine plots, either ungrazed or grazed by cattle, were selected. Ticks were collected weekly by blanket dragging. Borrelia infections were determined by PCR and restriction fragment length polymorphism. Rodent densities were estimated using mark-release-recapture methods. On occasion, the cattle were inspected for tick infestations. Meteorological data were recorded for each habitat. Significantly more ticks were collected in the ungrazed woodland than in the grazed woodland. The ungrazed oak habitat had higher tick densities than the pine habitat, while in the grazed habitats, tick densities were similar. Borrelia infection rates ranged from zero in larvae to 26% in nymphs to 33% in adult ticks, and B. afzelii, B. burgdorferi sensu stricto, B. garinii, and B. valaisiana were the species involved. Coinfections were found in five ticks. There was no effect of the presence of cattle on Borrelia infections in the ticks. In the ungrazed area, Borrelia infections in nymphs were significantly higher in the oak habitat than in the pine habitat. More mice were captured in the ungrazed area, and these had a significantly higher tick burden than mice from the grazed area. Tick burden on cattle was low. The results suggest that grazing has a negative effect on small rodents as well as on ticks but not on Borrelia infections. Implications of these results for management of woodland reserves and risk of Lyme disease are discussed.

Blackbirds (Turdus merula) and song thrushes (Turdus philomelos) were found to carry 95% of all spirochete-infected tick larvae among 40 bird species captured in Central Europe. More than 90% of the infections were typed as Borrelia garinii and Borrelia valaisiana. We conclude that thrushes are key players in the maintenance of these spirochete species in this region of Central Europe.

Verification of the efficacy of Biocan B inj. ad us. vet. (Bioveta, a.s.) was done by challenge testing. Ticks collected in the nature were used as natural vectors of the infection. Six beagles and two control ones were used in the test. Formation of outer surface protein A specific antibodies (OspA antibodies) and borrelia specific immonoglobulins (IgG) was measured by Western blot and EIA in the sera samples. The tissue samples were used for detection of borreliae by cultivation method and dark field microscopy (DFM). Formation of IgG antibodies and OspA antibodies after vaccination was observed. The maximum titer level of antibodies was reached between 21. and 49. day after vaccination and then slowly decreased. Presence of borreliae was detected only in skin biopsies of non-vaccinated dogs. The post mortem tissue samples showed presence of borreliae in all of the samples of the non-vaccinated dogs. The tissues of the vaccinated dogs were not infected with borreliae, except for two samples of dog with low titer levels of OspA antibodies. The development of the new vaccine is based on preparation of recombinant outer surface proteins (e.g. rOspA and rOspC) of B. afzelii, B. burgdorferi and B. garinii origin. Chosen recombinant proteins were successfully expressed in E. coli. The obtained purified proteins are currently being tested on laboratory BALB/c mice. Formation of specific antibodies against some recombinant proteins has been confirmed. These proteins are suitable candidates for preparation of a vaccine prototype and they will be subsequently used in challenge tests.

The prevalence of infection of Columbian black-tailed deer (Odocoileus hemionus columbianus) with Borrelia spp. was evaluated in an area of northwestern California (USA) where Lyme disease is endemic and the relapsing-fever group spirochete Borrelia coriaceae is enzootic, and in a far-removed comparison area having a disparate climate and lower density of vector ticks. Blood samples collected from both deer herds in 1987, 1988, and from 2000-02 were assayed for borrelial infection with microscopic and molecular methods. Serum specimens from two (5%) of 39 deer from the Dye Creek Preserve in Tehama County versus 13 (20%) of 64 animals from the Hopland Research and Extension Center (HREC) in Mendocino County, California were polymerase chain reaction (PCR) test positive for B. burgdorferi sensu lato. DNA sequencing analyses revealed that eight animals were infected with B. bissettii, six with three unclassified genotypes, and one with B. burgdorferi sensu stricto. One serum sample (2%) from HREC was positive for a relapsing-fever group spirochete that had a 16S rRNA sequence homology of 99% with the C053 type strain of B. coriaceae. Spirochetes undetermined to geno-species were detected in thick-blood drops prepared from three (8%) of 36 deer from the HREC by direct immunofluorescence. Adults of the hippoboscid flies Lipoptena depressa (n=73) and Neolipoptena ferrisi (n=24), the Pacific Coast tick (Dermacentor occidentalis) (n=22), and the western black-legged tick (Ixodes pacificus) (n=1) that had been removed from deer from both study areas in 2002 were PCR test negative for borreliae. The occurrence of diverse borreliae in deer from northern California confounds and, consequently, reduces the utility of borrelial serosurveys for detecting specific genospecies, unless they are complemented by more specific assays (e.g., immunoblotting, PCR/sequencing analysis).

The aim of this study was to improve understanding of several factors related to the ecology and environmental risk of Borrelia infection in northern Florida. Small mammals and host-seeking adult ticks were collected at several sites, and specimens were tested for the presence of Borrelia species, primarily by PCR amplification. Tissues from some vertebrates and ticks were initially cultured in BSK-H medium to isolate spirochetes, but none were recovered. However, comparison of partial flagellin (flaB), 66-kDa protein (p66), and outer surface protein A (ospA) gene sequences from DNAs amplified from small mammals and ticks confirmed the presence of several Borrelia species. Borrelia lonestari DNA was detected among lone star ticks (Amblyomma americanum) at four sites. Borrelia burgdorferi sensu stricto strains were detected in all small mammal species tested and in A. americanum, Ixodes affinis, and Ixodes scapularis ticks. Borrelia bissettii was found in a cotton mouse and cotton rats and in I. affinis ticks. The study findings extend the known geographic distributions of B. lonestari in A. americanum and of B. burgdorferi sensu lato in A. americanum, I. affinis, I. scapularis, and small mammals to new sites in Florida. The presence of B. burgdorferi sensu stricto strains in host-seeking lone star ticks at two sites in Florida suggests that A. americanum should still be considered a possible vector of B. burgdorferi sensu lato.

Some Borrelia species associated with Lyme disease bind the complement-regulatory protein factor H (fH), a process that may aid in immune evasion. In this report we demonstrate that some Borrelia species associated with relapsing fever bind fH, but not those associated with avian borreliosis and epizootic bovine abortion. Cell-bound fH was also found to mediate cleavage of exogenously supplied human C3b, demonstrating the biological relevance of fH binding and its possible importance in the pathogenesis of the relapsing-fever spirochetes.

To determine if white-tailed deer may serve as a reservoir host for Borrelia lonestari, we used a nested PCR for the Borrelia flagellin gene to evaluate blood samples collected from deer from eight southeastern states. Seven of 80 deer (8.7%) from 5 of 17 sites (29.4%) had sequence-confirmed evidence of a B. lonestari flagellin gene by PCR, indicating that deer are infected with B. lonestari or another closely related Borrelia species. Our findings expand the known geographic range of B. lonestari and provide the first evidence of this organism in a vertebrate other than humans.

Although Borrelia theileri, the agent of bovine borreliosis, was described at the turn of the century (in 1903), its relationship with borreliae causing Lyme disease or relapsing fever remains undescribed. We tested the previously published hypothesis that spirochetes infecting Lone Star ticks (Amblyomma americanum) may comprise B. theileri by analyzing the 16S ribosomal DNAs (rDNAs) and flagellin genes of these spirochetes. B. theileri, the Amblyomma agent, and B. miyamotoi formed a natural group or clade distinct from but most closely related to that of the relapsing fever spirochetes. B. theileri and the Amblyomma agent were 97 and 98% similar at the nucleotide level within the analyzed portions of the 16S rDNA and the flagellin gene respectively, suggesting a recent divergence. The agent of bovine borreliosis might be explored as a surrogate antigen for the as-yet-uncultivatable Amblyomma agent in studies designed to explore the etiology of a Lyme disease-like infection associated with Lone Star ticks.

Previous work described Borrelia burgdorferi sensu lato group DN127 as a new genospecies, Borrelia bissettii, and prompted the present study to identify the Borrelia spp. that exist in northern Colorado. To determine the genospecies present, we analyzed two specific intergenic spacer regions located between the 5S and 23S and the 16S and 23S ribosomal genes. Phylogenetic analysis of the derived sequences clearly demonstrated that these isolates, originating from rodents captured in the foothills of northern Colorado, diverged from B. burgdorferi sensu stricto by 5 to 5.5% and were members of the new genospecies B. bissettii.

Eleven pure cultures of Borrelia from 3 species of wild rodents (Apodemus agrarius, Mus formosanus, Rattus losea) captured in Taichung, located in the center of Taiwan island, and on Kinmen Island were characterized. Five isolates showed restriction fragment length polymorphism (RFLP) patterns of 5S-23S rRNA gene intergenic spacer sequences identical to those of strains 5MT and 10MT, identified as Borrelia valaisiana, which were isolated in the southern tip of South Korea. Although the remaining six isolates showed novel RFLP patterns, these isolates showed more similarity to members of B. valaisiana from Korea, Japan and Europe based on 16S rRNA gene and flagellin gene sequences. This led us to speculate that transmission and proliferation of this type of borrelia occurred between Taiwan and the southern part of South Korea.

During May 1996, field surveys on Lyme disease spirochetes were conducted in Beijing, Shenyang, Fushun, and Inner Mongolia in northeastern China. The ticks collected consisted of 3 genera and 12 species. Of these, Ixodes persulcatus was dominant in sun-exposed vegetation in forests in Inner Mongolia; 57 Borrelia strains (55/123 unfed adults and 2/5 immature stages fed on a rodent) were obtained from this tick by BSK culture. Additionally, 2/2 Apodemus peninsulae were positive. Ixodes nipponensis, Ixodes pavlovskyi, Haemaphysalis douglasi, and Haemaphysalis megaspinosa, newly recorded in China, and other Haemaphysalis spp. were all negative for Borrelia. Based on a polymerase chain reaction restriction fragment length polymorphism analysis of the 45 strains successfully subcultured, these were classified as 29 Borrelia garinii and 16 Borrelia afelii. These strains seemed to be more closely related to Japanese strains in genetic features than to those from Europe. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis suggested more diversity in both genospecies, but Borrelia burgdorferi sensu stricto was not found.

Spirochetes were isolated from the tick, Ixodes tanuki, as well as wood mice (Apodemus speciosus) and voles (Clethrionomys rufocanus and Eothenomys smithii), caught in Fukui, Tokushima, and Hokkaido, Japan, from 1991 to 1993. Spirochetes were characterized on the basis of protein profiles, reactivities with monoclonal antibodies (mAbs), Outer surface protein A gene (ospA) and Outer surface protein B gene (ospB) amplification analysis, rRNA gene and flagellin gene restriction fragment length polymorphism (RFLP) analysis, and DNA homology values. Protein profiles of all isolates were homologous and reacted with mAb to OspA, OspB, OspC, flagellin, and heat shock protein 60. The primer reactivity to ospA and ospB were different from those of Borrelia burgdorferi sensu stricto, B. afzelii, B. japonica, and B. garinii. Based on the DNA/DNA homology value and RFLP analysis of rRNA and flagellin gene, these Borrelia sp. isolates are a new group of B. burgdorferi sensu lato. The isolates from ticks and the host rodents were identical in these assays. We propose that these Borrelia sp. are adapted to I. tanuki and are maintained in these field rodents.

Objective: To determine the correlation of seroimmunologic test results between reference and nonreference laboratories., Design: Retrospective data analysis., Procedure: Serum samples obtained from naturally infected dogs and cats were distributed to reference and nonreference laboratories for seroimmunologic testing. Correlation of test results was evaluated by use of nonparametric analysis., Results: Correlation coefficients were high between laboratory groups for samples tested for feline immunodeficiency virus antibodies, FeLV antigen, and toxoplasmosis antibodies in cats. Results for feline immunodeficiency virus antibody tests from reference laboratories were more likely to be positive than results from nonreference laboratories. Test results for feline infectious peritonitis antibodies, antinuclear antibodies, and Borrelia antibodies in cats were not significant. Coefficient correlations were significant for results of heart-worm antigen, Brucella antibodies, Toxoplasma antibodies, antinuclear antibodies, and rheumatoid factor in dogs. Results for Borrelia antibodies were not correlated between laboratory groups., Clinical Implications: Results were highly correlated between reference and nonreference laboratories for 8 of 14 seroimmunologic tests. Seroimmunologic tests for use in cats were less correlated as a group than those for use in dogs. Poor correlation of results between laboratories was attributed to variations in control agents, antigens, reagents, technical expertise, and cutoff values and end-point titers used for diagnosis.

In autumn of 1994 and spring of 1995, we examined Borrelia infection among Microtinae voles, Clethrionomys rufocanus bedfordiae, in Hokkaido, Japan. In BSK culturing of the earlobe tissues of 45 C. rufocanus bedfordiae captured, twelve rodents were positive for Borrelia. Eight isolates were used for the polymerase chain reaction (PCR) and the restriction fragment length polymorphism (RFLP) analysis. According to the results, these isolates were classified into B. garinii or B. afzelii. It is considered that a common vole, C. rufocanus bedfordiae, plays a significant role in the transmission and maintenance of B. garinii and B. afzelii, similar to the role of Apodemus speciosus mice.

The morphology and ultrastructure of two strains of Borrelia anserina were investigated by electron microscopy of negatively stained and ultrathin sectioned cells. One was a cultivable strain originally isolated in the USA and the other was originally isolated in Nigeria and maintained in chickens. The cells were regularly helical, 9-21 microns long and 0.22-0.26 microns wide with a helix wavelength of about 1.7 microns. The cells were surrounded by a surface layer and appeared to divide by binary fission. The structure of the cells from each of the two strains was identical except that those of the USA strain possessed seven flagella inserted at each end and those from the Nigerian strain had eight.

Spirochetemia is a rarely reported observation in dogs. We describe the clinical, hematologic, and immunodiagnostic features of two spirochetemic dogs from northern Florida and the subsequent isolation and preliminary characterization of a Borrelia species from one dog in which culture of a sample for spirochetes was attempted. Results of polyacrylamide gel electrophoresis, monoclonal antibody testing, and PCR analysis indicate that the Florida isolate is not Borrelia burgdorferi, the only other member of the genus that has been isolated in Florida. Our findings also indicate that a member of the genus Borrelia potentially causes disease in dogs in Florida and that serologic cross-reactivity of the Florida canine Borrelia isolate with B. burgdorferi probably contributes to the inaccurate diagnosis of canine Lyme disease in the region.

An outbreak of spirochetosis occurred in a flock of 75 game chickens in California during fall 1991. Affected birds were weak and anemic. Many had greenish diarrhea. Spirochetes were seen on Giemsa-stained blood smears and in silver-stained tissue sections of kidney, liver, and spleen. Splenomegaly, which is reported to be characteristic of fowl spirochetosis, was not observed in two acutely infected chickens.

Six hundred sixty-five hunter-killed white-tailed deer (Odocoileus virginianus) from 18 counties in Alabama (USA) were examined for ticks. Most of the collections were made at state-operated wildlife management areas. Four species of ticks (n = 4,527) were recovered: the lone star tick Amblyomma americanum (n = 482); the Gulf Coast tick A. maculatum (n = 11); the winter tick Dermacentor albipictus (n = 1,242); and the black-legged tick Ixodes scapularis (n = 2,792). Fifty-six percent of the ticks (n = 2,555) were examined for Borrelia sp. spirochetes using an immunofluorescent, polyclonal antibody assay. Spirochetes were detected in I. scapularis (five females, seven males) from Barbour, Butler, Coosa, and Lee counties and A. americanum (four males, four nymphs) from Hale, Lee, and Wilcox counties. Area-specific prevalences in ticks were as high as 3.3% for I. scapularis and 3.8% for A. americanum.

A serologic survey for the agents of Rocky Mountain spotted fever (RMSF) (Rickettsia rickettsii), Borrelia spp. including the causative agent for Lyme disease (Borrelia burgdorferi), and plague (Yersinia pestis) was conducted on blood samples collected from 30 and 46 black-tailed jack rabbits (Lepus californicus) from an urban environment in Lubbock, Texas (USA) during winter 1987 and the following spring 1988, respectively. Antibody titers to the agents of RMSF and borreliosis were detected in sera of 28 and 1% of the jack rabbits, respectively. Neither organisms (rickettsiae and/or spirochetes) nor their associated antigens were detected in any of the tissue or whole blood samples; plague antibodies were not detected in the 76 jack rabbits sampled. Four of 18 ticks (Dermacentor parumapertus) removed from 12 jack rabbits were positive for RMSF using the fluorescent antibody test. The black-tailed jack rabbit is a common wildlife species living in close proximity to higher density human populations in many areas of the southwestern United States. Our results indicate the potential importance of urban populations of this mammal as reservoirs for at least one important zoonotic disease, RMSF, in western Texas.

Exposure of 10-week-old White Leghorn chickens to sheep blood containing Borrelia theileri neither resulted in infection nor conferred protection against B. anserina.

Blood, kidney and tick samples were obtained from 18 hunter-killed black bears (Ursus americanus) from three sites in northern Wisconsin. A Borrelia sp., morphologically and antigenically similar to Borrelia burgdorferi, was isolated from the blood of two of the animals, and from the kidney of a third. Ixodes dammini and Dermacentor variabilis were found on the bears. This is the first report of borreliosis in the Ursidae, and of the primary vector of Lyme disease, I. dammini, from this host.

Borrelia burgdorferi, the causative agent of Lyme disease, was isolated from 15 of 17 white-footed mice (Peromyscus leucopus) and 54 of 82 subadult Ixodes dammini from Fort McCoy, Wis. Of the 47 isolates tested, all reacted in indirect fluorescent-antibody tests with monoclonal antibodies directed against a surface protein of B. burgdorferi (approximate molecular weight, 31,000) and flagellins that are common to all Borrelia species. Indirect fluorescent-antibody reactions were variable when an antibody that binds to a surface protein with an approximate molecular weight of 34,000 was tested. The major proteins of isolates from ticks and mice had approximate molecular weights of 31,000, 34,000, and 41,000. Antibodies to B. burgdorferi were present (titer, greater than or equal to 1:64) in 16 of 97 white-tailed deer (Odocoileus virginianus). The mean number of subadult I. dammini on mice captured in June 1986 was 6.5, and the mean number of adult I. dammini on deer killed in November 1986 was 5.9. The presence of ticks and the high prevalence of I. dammini and mice infected with B. burgdorferi establish that Fort McCoy is an area in which the Lyme disease agent is highly endemic, even though there have been relatively few documented cases in humans. The low number of reported cases in humans may be a result of National Guard and reserve unit personnel returning home to civilian life and having symptoms expressed subsequently, or it could be due to misdiagnosis or nonreporting.

This investigation defined the extent of Borrelia burgdorferi infection surrounding La Crosse, Wis. White-footed mice, Peromyscus leucopus or P. maniculatis, were captured from sites in Wisconsin, Minnesota, and Iowa and cultured for B. burgdorferi to define the local boundaries of the midwestern Lyme disease area. All foci of B. burgdorferi infection (N1, N2, N3, and N4) were located north of interstate highway 90 except focus S2, which was south of the highway near Fort McCoy, Wis. The interstate highway may have been a barrier to deer movement which slowed the southward dispersal of Ixodes dammini. B. burgdorferi was isolated from 12 (63%) of the mice captured from site N4, which was adjacent to the western border of Fort McCoy. Unexpectedly, no B. burgdorferi-infected mice were isolated at site N0, located north of interstate highway 90 and enclosed by areas in which B. burgdorferi infection is endemic. This site is surrounded by natural barriers which may have slowed the spread of I. dammini by deer. The Wisconsin area in which B. burgdorferi is endemic should now include the surrounding area north of interstate highway 90 west from Fort McCoy to the Mississippi River. Additional studies are needed to define the rapidity, limits, and means of I. dammini dispersal into southern Wisconsin.

This study evaluated the differences in immunoblot patterns when serum specimens from seropositive dogs were reacted against four strains of Borrelia burgdorferi. Intense bands were consistently detected for the 83-, 66-, 61- to 60-, 41-, and 31-kilodalton regions for all four strains. Most variations were observed in the regions of 45 to 34 and 26 to 15 kilodaltons. Adsorption studies suggested that one reason for the observed variability was a difference in proteins among the organisms, rather than a difference in migration of proteins. Therefore, knowledge and consistency of the test antigen are essential when evaluating and comparing canine immunoblot patterns to B. burgdorferi, but for diagnostic purposes all of the serum samples would have been considered positive regardless of the strain used.

In northern California, antibodies to Borrelia burgdorferi were detected in 58 of 73 (79%), and spirochetemias in one of 26 (4%) black-tailed jackrabbits (Lepus californicus californicus), by indirect and direct immunofluorescence, respectively. Five species of ticks (Dermacentor occidentalis, D. parumapertus, Ixodes neotomae, I. pacificus, and Haemaphysalis leporispalustris) were collected from rabbits. Two of these species of ticks were found to contain spirochetes; two of 10 (20%) I. neotomae and two of 174 (1%) H. leporispalustris. A strain of B. burgdorferi was recovered from I. neotomae. One infected H. leporispalustris female passed spirochetes via eggs to about 67% of her progeny. The widespread distribution of the black-tailed jackrabbit, its infestation by at least four ticks (D. occidentalis, D. parumapertus, I. neotomae, and I. pacificus) known to be infected naturally with B. burgdorferi, and the high prevalence of spirochetal antibody in this lagomorph suggest that it might be useful as a sentinel for surveillance of Lyme borreliosis. Spirochetes were detected in 15% of 40 Columbian black-tailed deer (Odocoileus hemionus columbianus) by direct immunofluorescence bound with a Borrelia-specific monoclonal antibody (H9724), but not with a monoclonal antibody (H5332) specific for B. burgdorferi. The geographical overlap of different borreliae in ticks that bite wildlife such as deer may confound spirochetal serosurveys, and underscores the need for more specific serologic tests than those currently available.

Borrelia burgdorferi, the etiologic agent of Lyme disease, was isolated from 111 of 237 Peromyscus leucopus captured during all seasons of the year. Borreliae were cultured from tissues of the spleen (101 mice), left kidney (76 mice), and right kidney (73 mice), from blood (12 mice), and from one fetus. Mice were infected during the winter, when immature Ixodes dammini were inactive. The prevalence of infection during the winter (less than or equal to 33%) was more than twofold lower than that during the summer (ca. 75%), a time when nymphal ticks are abundant. Overwintering, infected mice are reservoir hosts for subadult ticks that begin feeding in early spring. Twenty white-footed mice from which B. burgdorferi was isolated from tissues of spleen or kidney but not from blood were parasitized by larval I. dammini or Dermacentor variabilis which harbored borreliae. We conclude that these mice were infectious to feeding ticks, even though borreliae were not isolated from blood.

Infectious bursal disease and spirochetosis were diagnosed simultaneously in a flock of 100 six-week-old babcock pullet chicks. Larvae of the fowl tick Argas persicus were found on the bodies of the chicks. Thirty-nine of the chicks died 5 days after the onset of clinical signs.