Summary Background Lyme borreliosis is the most common tick-borne disease in the northern hemisphere. It is a multisystem disease caused by Borrelia burgdorferi sensu lato genospecies and characterised by tissue localisation and low spirochaetaemia. In this study we aimed to describe a novel Borrelia species causing Lyme borreliosis in the USA. Methods At the Mayo clinic, from 2003 to 2014, we tested routine clinical diagnostic specimens from patients in the USA with PCR targeting the oppA1 gene of B burgdorferi sensu lato. We identified positive specimens with an atypical PCR result (melting temperature outside of the expected range) by sequencing, microscopy, or culture. We collected Ixodes scapularis ticks from regions of suspected patient tick exposure and tested them by oppA1 PCR. Findings 100 545 specimens were submitted by physicians for routine PCR from Jan 1, 2003 to Sept 30, 2014. From these samples, six clinical specimens (five blood, one synovial fluid) yielded an atypical oppA1 PCR product, but no atypical results were detected before 2012. Five of the six patients with atypical PCR results had presented with fever, four had diffuse or focal rash, three had symptoms suggestive of neurological inclusion, and two were admitted to hospital. The sixth patient presented with knee pain and swelling. Motile spirochaetes were seen in blood samples from one patient and cultured from blood samples from two patients. Among the five blood specimens, the median oppA1 copy number was 180 times higher than that in 13 specimens that tested positive for B burgdorferi sensu stricto during the same time period. Multigene sequencing identified the spirochaete as a novel B burgdorferi sensu lato genospecies. This same genospecies was detected in ticks collected at a probable patient exposure site. Interpretation We describe a new pathogenic Borrelia burgdorferi sensu lato genospecies (candidatus Borrelia mayonii) in the upper midwestern USA, which causes Lyme borreliosis with unusually high spirochaetaemia. Clinicians should be aware of this new B burgdorferi sensu lato genospecies, its distinct clinical features, and the usefulness of oppA1 PCR for diagnosis.

Hard ticks of the order Ixodidae serve as vectors for numerous human pathogens, including the causative agent of Lyme Disease Borrelia burgdorferi. Tick-associated microbes can influence pathogen colonization, offering the potential to inhibit disease transmission through engineering of the tick microbiota. Here, we investigate whether B. burgdorferi encounters abundant bacteria within the midgut of wild adult Ixodes scapularis, its primary vector. Through the use of controlled sequencing methods and confocal microscopy, we find that the majority of field-collected adult I. scapularis harbor limited internal microbial communities that are dominated by endosymbionts. A minority of I. scapularis ticks harbor abundant midgut bacteria and lack B. burgdorferi. We find that the lack of a stable resident midgut microbiota is not restricted to I. scapularis since extension of our studies to I. pacificus, Amblyomma maculatum, and Dermacentor spp showed similar patterns. Finally, bioinformatic examination of the B. burgdorferi genome revealed the absence of genes encoding known interbacterial interaction pathways, a feature unique to the Borrelia genus within the phylum Spirochaetes. Our results suggest that reduced selective pressure from limited microbial populations within ticks may have facilitated the evolutionary loss of genes encoding interbacterial competition pathways from Borrelia.

Lyme borreliosis (LB) is a multisystem disease caused by spirochetes in the Borrelia burgdorferi sensu lato (Bbsl) genospecies complex. We previously described a novel Bbsl genospecies (type strain MN14-1420T) that causes LB among patients with exposures to ticks in the upper midwestern USA. Patients infected with the novel Bbsl genospecies demonstrated higher levels of spirochetemia and somewhat differing clinical symptoms as compared with those infected with other Bbsl genospecies. The organism was detected from human specimens using PCR, microscopy, serology and culture. The taxonomic status was determined using an eight-housekeeping-gene (uvrA, rplB, recG, pyrG, pepX, clpX, clpA and nifS) multi-locus sequence analysis (MLSA) and comparison of 16S rRNA gene, flaB, rrf–rrl, ospC and oppA2 nucleotide sequences. Using a system threshold of 98.3% similarity for delineation of Bbsl genospecies by MLSA, we demonstrated that the novel species is a member of the Bbsl genospecies complex, most closely related to B. burgdorferi sensu stricto (94.7–94.9% similarity). This same species was identified in Ixodes scapularis ticks collected in Minnesota and Wisconsin. This novel species, Borrelia mayonii sp. nov, is formally described here. The type strain, MN14-1420, is available through the Deutsche Sammlung von Mikroorganismen und Zelkulturen GmbH (DSM 102811) and the American Type Culture Collection (ATCC BAA-2743).

As a result of the increasing incidence of Lyme disease and other tick-borne pathogens in Wisconsin, we assessed the distribution of adult blacklegged ticks through collections from hunter-killed deer in 2008 and 2009 and compared results with prior surveys beginning in 1981. Volunteers staffed 21 Wisconsin Department of Natural Resources registration stations in 21 counties in the eastern half of Wisconsin in 2008 and 10 stations in seven counties in northwestern Wisconsin in 2009. In total, 786 and 300 white-tailed deer (Odocoileus virginianus) were examined in 2008 and 2009, respectively. All but three stations in 2008 were positive for ticks and all stations in 2009 were positive for ticks. The three sites negative for ticks occurred within the eastern half of Wisconsin. The results indicate that range expansion of Ixodes scapularis (Say) is continuing and the risk of tick exposure is increasing, especially in the eastern one-third of the state.

MHz conductivity, torque magnetometer and magnetization measurements are reported on single crystals of CeOs4Sb12 and NdOs4Sb12 using temperatures down to 0.5 K and magnetic fields of up to 60 tesla. The field-orientation dependence of the de Haas-van Alphen and Shubnikov-de Haas oscillations is deduced by rotating the samples about the [010] and [011] directions. The results indicate that NdOs4Sb12 has a similar Fermi surface topology to that of the unusual superconductor PrOs4Sb12, but with significantly smaller effective masses, supporting the importance of local phonon modes in contributing to the low-temperature heat capacity of NdOs4Sb12. By contrast, CeOs4Sb12 undergoes a field-induced transition from an unusual semimetal into a high-field, hightemperature state characterized by a single, almost spherical Fermi-surface section. The behavior of the phase boundary and comparisons with models of the bandstructure lead us to propose that the field-induced phase transition in CeOs4Sb12 is similar in origin to the well-known α − γ transition in Ce and its alloys.

Two species of mice, the white-footed mouse, Peromyscus leucopus (Rafinesque; Rodentia: Cricetidae) and the woodland deer mouse, Peromyscus maniculatus (Wagner; Rodentia: Cricetidae), serve as reservoirs of tick-borne pathogens in many parts of North America. However, the role P. maniculatus plays in the amplification and maintenance of Anaplasma phagocytophilum (Rickettsiales: Ehrlichiaceae) and Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) is not well understood. In northern Wisconsin, from 2012 to 2014, 560 unique mice were captured at 83 sites distributed across five forests. P. leucopus was more likely infested with immature Ixodes scapularis compared to P. maniculatus (60.1 vs. 28.3%). Abundance of immature I. scapularis on P. leucopus (M = 2.69; SD = 3.53) was surprisingly low and even lower for P. maniculatus (M = 0.717; SD = 1.44). Both P. leucopus and P. maniculatus were infected with B. burgdorferi, 24.0 and 16.8%, respectively. The prevalence of A. phagocytophilum infection in P. leucopus (1.69%) was similar to that observed in P. maniculatus (4.73%). Nine of 10 mice co-infected with both pathogens were P. maniculatus, and there were more co-infections in this species than expected by chance (3.07 vs. 0.82%). Differences in the behavior and biology between these two mice species may contribute to the variation observed in the abundance of host-attached ticks and pathogen prevalence. These differences highlight a potential hazard of the failure to differentiate between these visually similar mice, but there is evidence that these two mice species can each serve as reservoirs for tick-borne pathogens that cause human disease in Wisconsin.

1Hard ticks of the order Ixodidae serve as vectors for numerous human pathogens, including the 2 causative agent of Lyme Disease Borrelia burgdorferi. Tick-associated microbes can influence 3 pathogen colonization, offering the potential to inhibit disease transmission through engineering 4 of the tick microbiota. Here, we investigate whether B. burgdorferi encounters abundant bacteria 5 within the midgut of wild adult Ixodes scapularis, its primary vector. Through the use of 6 controlled sequencing methods and confocal microscopy, we find that the majority of field-7 collected adult I. scapularis harbor limited internal microbial communities that are dominated by 8 endosymbionts. A minority of I. scapularis ticks harbor abundant midgut bacteria and lack B. 9 burgdorferi. We find that the lack of a stable resident midgut microbiota is not restricted to I. 10 scapularis since extension of our studies to I. pacificus, Amblyomma maculatum, and 11Dermacentor spp showed similar patterns. Finally, bioinformatic examination of the B. 12 burgdorferi genome revealed the absence of genes encoding known interbacterial interaction 13 pathways, a feature unique to the Borrelia genus within the phylum Spirochaetes. Our results 14 suggest that reduced selective pressure from limited microbial populations within ticks may have 15 facilitated the evolutionary loss of genes encoding interbacterial competition pathways from 16Borrelia.

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