Borrelia miyamotoi sensu lato Seroreactivity and Seroprevalence in the Northeastern United States

Borrelia miyamotoi sensu lato, a relapsing fever Borrelia sp., is transmitted by the same ticks that transmit B. burgdorferi (the Lyme disease pathogen) and occurs in all Lyme disease�?"endemic areas of the United States. To determine the seroprevalence of IgG against B. miyamotoi sensu lato in the northeastern United States and assess whether serum from B. miyamotoi sensu lato�?"infected persons is reactive to B. burgdorferi antigens, we tested archived serum samples from area residents during 1991�?"2012. Of 639 samples from healthy persons, 25 were positive for B. miyamotoi sensu lato and 60 for B. burgdorferi. Samples from �%^10% of B. miyamotoi sensu lato�?"seropositive persons without a recent history of Lyme disease were seropositive for B. burgdorferi. Our resultsA suggest thatA human B. miyamotoiA sensu latoA infection may be common in southern New England and that B. burgdorferi antibody testing is not an effective surrogate for detecting B. miyamotoi sensu lato infection.     

我国莱姆病的流行病学和病原学研究

１９８７－１９９６年对２２个省市（市、区）的６０个县、区进行了莱姆病的调查研究。血清流行病学证实２２个省市（市、区）林区人群均存在莱姆病感染，感染率平均为５．０６％（１７２４／３４１０４）。病原学证实１７个省（市、区）存在莱姆病自然疫源地。１１个省（市、区）有典型病例存在。     

Evolution of Northeastern and Midwestern Borrelia burgdorferi, United States

The per capita incidence of human Lyme disease in the northeastern United States is more than twice that in the Midwest. However, the prevalence of Borrelia burgdorferi, the bacterium that causes Lyme disease, in the tick vector is nearly identical in the 2 regions. The disparity in human Lyme disease incidence may result from a disparity in the human invasiveness of the bacteria in the Northeast and Midwest caused by fundamentally different evolutionary histories. B. burgdorferi populations in the Northeast and Midwest are geographically isolated, enabling evolutionary divergence in human invasiveness. However, we found that B. burgdorferi populations in the Northeast and Midwest shared a recent common ancestor, which suggests that substantial evolutionary divergence in human invasiveness has not occurred. We propose that differences in either animal ecology or human behavior are the root cause of the differences in human incidence between the 2 regions.     

Infections with Tickborne Pathogens after Tick Bite,Austria, 2015–2018

The aim of this prospective study was to assess the risk for tickborne infections after a tick bite. A total of 489 persons bitten by 1,295 ticks were assessed for occurrence of infections with Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia spp., Babesia spp., Candidatus Neoehrlichia mikurensis, and relapsing fever borreliae. B. burgdorferi s.l. infection was found in 25 (5.1%) participants, of whom 15 had erythema migrans. Eleven (2.3%) participants were positive by PCR for Candidatus N. mikurensis. One asymptomatic participant infected with B. miyamotoi was identified. Full engorgement of the tick (odds ratio 9.52) and confirmation of B. burgdorferi s.l. in the tick by PCR (odds ratio 4.39) increased the risk for infection. Rickettsia helvetica was highly abundant in ticks but not pathogenic to humans. Knowledge about the outcome of tick bites is crucial because infections with emerging pathogens might be underestimated because of limited laboratory facilities.     

Evolutionary genomics of Borrelia burgdorferi sensu lato: Findings,hypotheses, and the rise of hybrids

Borrelia burgdorferi sensu lato (B. burgdorferi s.l.), the group of bacterial species represented by Lyme disease pathogens, has one of the most complex and variable genomic architectures among prokaryotes. Showing frequent recombination within and limited gene flow among geographic populations, the B. burgdorferi s.l. genomes provide an excellent window into the processes of bacterial evolution at both within- and between-population levels. Comparative analyses of B. burgdorferi s.l. genomes revealed a highly dynamic plasmid composition but a conservative gene repertoire. Gene duplication and loss as well as sequence variations at loci encoding surface-localized lipoproteins (e.g., the PF54 genes) are strongly associated with adaptive differences between species. There are a great many conserved intergenic spacer sequences that are candidates for cis-regulatory elements and non-coding RNAs. Recombination among coexisting strains occurs at a rate approximately three times the mutation rate. The coexistence of a large number of genomic groups within local B. burgdorferi s.l. populations may be driven by immune-mediated diversifying selection targeting major antigen loci as well as by adaptation to multiple host species. Questions remain regarding the ecological causes (e.g., climate change, host movements, or new adaptations) of the ongoing range expansion of B. burgdorferi s.l. and on the genomic variations associated with its ecological and clinical variability. Anticipating an explosive growth of the number of B. burgdorferi s.l. genomes sampled from both within and among species, we propose genome-based methods to test adaptive mechanisms and to identify molecular bases of phenotypic variations. Genome sequencing is also necessary for monitoring a likely increase of genetic admixture of previously isolated species and populations in North America and elsewhere.     

Failure of the Asian longhorned tick,Haemaphysalis longicornis,to serve as an experimental vector of the Lyme disease spirochete,Borrelia burgdorferi sensu stricto

The invasive, human-biting Asian longhorned tick, Haemaphysalis longicornis, was detected in New Jersey in the eastern United States in August of 2017 and by November of 2018 this tick had been recorded from 45 counties across 9 states, primarily along the Eastern Seaboard. The establishment of H. longicornis in the United States has raised the questions of how commonly it will bite humans and which native pathogens may naturally infect this tick. There also is a need for experimental vector competence studies with native pathogens to determine if H. longicornis can acquire a given pathogen while feeding, pass it transstadially, and then transmit the pathogen in the next life stage. In this experimental study, we evaluated the vector competence of a population of H. longicornis originating from the United States (New York) for a native isolate (B31) of the Lyme disease spirochete, Borrelia burgdorferi sensu stricto (s.s.). In agreement with a previous experimental study on the vector competence of H. longicornis for Borrelia garinii, we found that uninfected H. longicornis larvae could acquire B. burgdorferi s.s. while feeding on infected Mus musculus mice (infection prevalence >50% in freshly fed larvae) but that the infection was lost during the molt to the nymphal stage. None of 520 tested molted nymphs were found to be infected, indicating that transstadial passage of B. burgdorferi s.s. is absent or rare in H. longicornis; and based on the potential error associated with the number of nymphs testing negative in this study, we estimate that the upper 95% limit for infection prevalence was 0.73%. An Ixodes scapularis process control showed both effective acquisition of B. burgdorferi s.s. from infected mice by uninfected larvae and transstadial passage to the nymphal stage (infection prevalence of 80–82% for both freshly fed larvae and molted nymphs). We also observed that although H. longicornis larvae could be compelled to feed on mice by placing the ticks within feeding capsules, attachment and feeding success was minimal (<0.5%) when larvae were placed freely on the fur of the mice. We conclude that H. longicornis is unlikely to contribute more than minimally, if at all, to transmission of Lyme disease spirochetes in the United States.     

Lyme Disease,Virginia, USA, 2000–2011

Lyme disease, caused by the bacterium Borrelia burgdorferi and transmitted in the eastern United States by the black-legged tick (Ixodes scapularis), is increasing in incidence and expanding geographically. Recent environmental modeling based on extensive field collections of host-seeking I. scapularis ticks predicted a coastal distribution of ticks in mid-Atlantic states and an elevational limit of 510 m. However, human Lyme disease cases are increasing most dramatically at higher elevations in Virginia, a state where Lyme disease is rapidly emerging. Our goal was to explore the apparent incongruity, during 2000–2011, between human Lyme disease data and predicted and observed I. scapularis distribution. We found significantly higher densities of infected ticks at our highest elevation site than at lower elevation sites. We also found that I. scapularis ticks in Virginia are more closely related to northern than to southern tick populations. Clinicians and epidemiologists should be vigilant in light of the changing spatial distributions of risk.     

Current Guidelines,Common Clinical Pitfalls,and Future Directions for Laboratory Diagnosis of Lyme Disease,United States

In the United States, Lyme disease is caused by Borrelia burgdorferi and transmitted to humans by blacklegged ticks. Patients with an erythema migrans lesion and epidemiologic risk can receive a diagnosis without laboratory testing. For all other patients, laboratory testing is necessary to confirm the diagnosis, but proper interpretation depends on symptoms and timing of illness. The recommended laboratory test in the United States is 2-tiered serologic analysis consisting of an enzyme-linked immunoassay or immunofluorescence assay, followed by reflexive immunoblotting. Sensitivity of 2-tiered testing is low (30%–40%) during early infection while the antibody response is developing (window period). For disseminated Lyme disease, sensitivity is 70%–100%. Specificity is high (>95%) during all stages of disease. Use of other diagnostic tests for Lyme disease is limited. We review the rationale behind current US testing guidelines, appropriate use and interpretation of tests, and recent developments in Lyme disease diagnostics.     

Antibodies against Borrelia burgdorferi sensu lato among Adults,Germany, 2008–2011

To assess Borrelia burgdorferi sensu lato (the cause of Lyme borreliosis) seropositivity in Germany, we tested serum samples from health survey (2008–2011) participants. Seroprevalence was 5.8% among women and 13.0% among men; infection risk was highest among persons >60 years of age. Public health interventions, including education about risk factors and preventive measures, are needed.Key words: Lyme borreliosis, Borrelia burgdorferi sensu lato, seroepidemiologic studies, adults, Germany, risk factors, seroprevalence, prevalence, tickborne disease, bacteria, Ixodes ricinus, Ixodes persulcatusLyme borreliosis, the most common tickborne disease in the Northern Hemisphere, is caused by infection with spirochetes of the Borrelia burgdorferi sensu lato (s.l.) complex. Five genospecies are known to be pathogenic for humans: B. burgdorferi sensu stricto (s.s.), B. afzelii, B. garinii, B. bavariensis, and B. spielmanii (1). In Europe, the bacterium is transmitted to humans through the bite of Ixodes ricinus ticks; in eastern Europe, I. persulcatus ticks can also transmit the bacterium.In Europe, where the most common clinical manifestation of Lyme borreliosis is erythema migrans, followed by Lyme neuroborreliosis and Lyme arthritis (2), data are sparse regarding B. burgdorferi s.l. infection rates and risk factors (3). Persons of all ages are at risk for infection; however, surveillance data and prospective studies in Europe and the United States suggest that children and the elderly are particularly at risk (4–6). Population-based surveillance data suggest that Lyme borreliosis is endemic in eastern Germany: annual incidence is 20–35 cases/100,000 inhabitants (7). Regional differences in incidence are observed, but data cannot be easily compared because of reporting biases and differences in infection awareness.The limited representativeness and comparability of Lyme borreliosis surveillance data are well documented (8). Under such conditions, population-based serosurveys with high representativeness can provide valid estimates of the force of infection (rate at which susceptible persons acquire Lyme borreliosis) and the lifetime risk for infection; however, seroprevalence estimates do not necessarily represent cases of clinical disease. In a population-based seroprevalence study among 1- to 17-year-old children in Germany, seroprevalence increased cumulatively by age (9). We present data on the prevalence and determinants of B. burgdorferi s.l. seropositivity among adults in Germany during 2008–2011.     

Values of diagnostic tests for the various species of spirochetes

Bacteria of the Borrelia burgdorferi sensu lato complex, responsible for Lyme disease, are members of the spirochetes phylum. Diagnostic difficulties of Lyme disease are partly due to the characteristics of spirochetes as their culture is tedious or even impossible for some of them. We performed a literature review to assess the value of the various diagnostic tests of spirochetes infections of medical interest such as Lyme borreliosis, relapsing fever borreliae, syphilis, and leptospirosis. We were able to draw similarities between these four infections. Real-time PCR now plays an important role in the direct diagnosis of these infections. However, direct diagnosis remains difficult because of a persistent lack of sensitivity. Serological testing is therefore crucial in the diagnostic process. All currently available diagnostic tools are imperfect, with a potential risk of false positive and false negative results depending on the clinical context. Physicians should always take into consideration the clinical and epidemiological context when Lyme disease, relapsing fever borreliae, syphilis, and leptospirosis are suspected.     

Mapping distributions of the Lyme disease vector,Ixodes scapularis,and spirochete,Borrelia burgdorferi,in Kentucky using passive and active surveillance

Lyme disease is the most common tick-borne illness in the United States and is becoming more prevalent each year. It is transmitted to humans and animals through the bites of Ixodes scapularis ticks infected with Borrelia burgdorferi in the eastern United States, I. pacificus in the western U.S, and I. ricinus in Europe and Asia. In Kentucky, where Lyme disease is non-endemic, the number of reported human cases in 2010 totaled five. In 2019, that number had increased by over 300%. Identifying the distribution of I. scapularis populations infected with B. burgdorferi is important data for effective prevention strategies and an important first step in monitoring disease spread. In collaboration with the Kentucky Department for Public Health, we performed surveillance for I. scapularis throughout the state of Kentucky using both active and passive surveillance methods. Diagnostic testing for the identification of B. burgdorferi (sensu stricto) was also conducted. We identified 457 I. scapularis ticks from March 2019 to December 2020 from 32 counties in Kentucky. B. burgdorferi was detected in I. scapularis populations collected from 14 different counties. These results add to the little data that exists in Kentucky on I. scapularis and B. burgdorferi distribution.     

Active and Passive Surveillance and Phylogenetic Analysis of Borrelia burgdorferi Elucidate the Process of Lyme Disease Risk Emergence in Canada

Background

Northward expansion of the tick Ixodes scapularis is driving Lyme disease (LD) emergence in Canada. Information on mechanisms involved is needed to enhance surveillance and identify where LD risk is emerging.

Objectives

We used passive and active surveillance and phylogeographic analysis of Borrelia burgdorferi to investigate LD risk emergence in Quebec.

Methods

In active surveillance, we collected ticks from the environment and from captured rodents. B. burgdorferi transmission was detected by serological analysis of rodents and by polymerase chain reaction assays of ticks. Spatiotemporal trends in passive surveillance data assisted interpretation of active surveillance. Multilocus sequence typing (MLST) of B. burgdorferi in ticks identified likely source locations of B. burgdorferi.

Results

In active surveillance, we found I. scapularis at 55% of sites, and we were more likely to find them at sites with a warmer climate. B. burgdorferi was identified at 13 I. scapularis–positive sites, but infection prevalence in ticks and animal hosts was low. Low infection prevalence in ticks submitted in passive surveillance after 2004—from the tick-positive regions identified in active surveillance—coincided with an exponential increase in tick submissions during this time. MLST analysis suggested recent introduction of B. burgdorferi from the northeastern United States.

Conclusions

These data are consistent with I. scapularis ticks dispersed from the United States by migratory birds, founding populations where the climate is warmest, and then establishment of B. burgdorferi from the United States several years after I. scapularis have established. These observations provide vital information for public health to minimize the impact of LD in Canada.     

Borrelia burgdorferi hijacks cellular metabolism of immune cells: Consequences for host defense

Changes in cellular metabolism have proven to be important factors in driving cell behavior. It has been shown that cellular metabolism of immune cells changes when exposed to or infected by several pathogens: while this is often an adaptation of the host cells to the infection, sometimes it represents a mechanism through which the pathogens evade immune activation. Borrelia burgdorferi sensu lato, the causative agent of Lyme borreliosis, is a pathogen that highly depends on the host to survive, as the bacterium lacks many central metabolic pathways to generate its own nutrients. It is therefore quite likely that the bacterium interacts with host cells to obtain these metabolites and thereby affects metabolism in the host. Previously, several studies have assessed metabolic pathways in B. burgdorferi s.l. and how it adapts to its different host species. However, few studies have looked into how the interaction with the bacterium might affect the host cell metabolism. In this review we present the major metabolic pathways activated during Lyme borreliosis, viewed from both bacterium and host metabolism, and we discuss how these pathways interact with each other, and how they influence pathogenesis of Lyme borreliosis.     

山东省莱姆病地理流行病学研究

目的调查山东省莱姆病地理分布、媒介生物、动物宿主及病原分离情况,为今后防治提供依据。方法采用间接免疫荧光抗体试验(IFA)对6个调查点的人群进行莱姆病抗体检测;用直接免疫荧光抗体试验(DFA)检查优势蜱种及其携带莱姆病螺旋体情况,并用BSKⅡ培养基分离培养莱姆病螺旋体。宿主调查采用DFA检查鼠肾。用单克隆抗体和IFA进行菌株的鉴定。结果共检测血清1934份,阳性121份,感染率6.26%;鲁东、鲁南、鲁中、鲁西北感染率分别为5.76%、7.03%、0、9.81%;各年龄组差异无统计学意义(P>0.05);男性感染率为5.56%,女性为6.98%,两者间差异无统计学意义(χ2=1.67,P>0.05);山东省蜱类的优势种为长角血蜱,该蜱中肠带螺旋体率为12.00%;宿主动物调查显示,鼠类带莱姆病螺旋体率为13.26%,黑线姬鼠为优势种(占45.65%),带菌率高为14.24%。从86组培养物中分离到2株莱姆病螺旋体(TSH1、TSH2)。结论不同地区生态环境和传播媒介分布不同,各地区间莱姆病感染程度差别很大。长角血蜱可能为山东省莱姆病的重要生物媒介。     

Humans infected with relapsing fever spirochete Borrelia miyamotoi, Russia

Borrelia miyamotoi is distantly related to B. burgdorferi and transmitted by the same hard-body tick species. We report 46 cases of B. miyamotoi infection in humans and compare the frequency and clinical manifestations of this infection with those caused by B. garinii and B. burgdorferi infection. All 46 patients lived in Russia and had influenza-like illness with fever as high as 39.5°C; relapsing febrile illness occurred in 5 (11%) and erythema migrans in 4 (9%). In Russia, the rate of B. miyamotoi infection in Ixodes persulcatus ticks was 1%-16%, similar to rates in I. ricinus ticks in western Europe and I. scapularis ticks in the United States. B. miyamotoi infection may cause relapsing fever and Lyme disease-like symptoms throughout the Holarctic region of the world because of the widespread prevalence of this pathogen in its ixodid tick vectors.     

Geodemographic analysis of Borrelia burgdorferi sensu lato using the 5S–23S rDNA spacer region

BackgroundLyme borreliosis is the predominant tick-borne disease in the Northern hemisphere, with considerable heterogeneity in clinical manifestations. Here, we evaluated one genetic marker for its use in population genetic based analysis. For that we collected molecular and epidemiological records of Borrelia burgdorferi sensu lato isolates from ticks, animals and humans at various sites in The Netherlands and worldwide.MethodsThe 5S–23S rDNA (rrfA–rrlB) intergenic spacer region (IGS) from 291 Dutch Borrelia positive ticks was sequenced and compared to Borrelia sequences from GenBank. We estimated several population genetic measures to test the neutrality of the marker. We also assessed the ability of this marker to discriminate between Eurasian Borrelieae at a finer geographical resolution, and to detect population expansion per genospecies.ResultsThe most prevalent genospecies in The Netherlands was Borrelia afzelii, whereas Borrelia garinii, B. burgdorferi sensu stricto, Borrelia spielmanii and Borrelia valaisiana were found less frequently. The result of the Ewens–Watterson–Slatkin test was consistent with neutral selection of IGS region. Estimated pairwise fixation indices (Fst) were significantly different from zero between The Netherlands, the rest of Europe, Russia and Asia for B. afzelii and Borrelia garinii. Estimated Fu’s Fs were significantly negative for B. afzelii and B. garinii.ConclusionsAt least seven B. burgdorferi s.l. genospecies circulate in Ixodes ricinus population in The Netherlands. The population genetic analyses of IGS region can resolve subpopulations within a genospecies and detect a large excess of rare genetic variants at the genospecies level. A genetic trace of population expansion for B. afzelii and B. garinii is consistent with the reported increase in Lyme borreliosis incidence in European countries.     

Effects of cattle grazing on Ixodes ricinus-borne disease risk in forest areas of the Netherlands

Cattle grazing has been suggested to reduce the risk for Lyme borreliosis by decreasing the density of questing Ixodes ricinus infected with Borrelia burgdorferi sensu lato. We tested the hypotheses that cattle grazing used in woodland management decreases the density of questing I. ricinus, and that it decreases the nympal infection prevalence of B. burgdorferi sensu lato. We further expected the nympal infection prevalence of tick-borne pathogens that utilize cattle as amplifying hosts, namely Anaplasma phagocytophilum, and Babesia sensu stricto, to increase. To test these hypotheses, we compared the densities of questing I. ricinus between twenty pairs of plots in grazed and ungrazed forest areas. The density of I. ricinus adults, but not nymphs, was lower in areas grazed by cattle than in ungrazed areas. Nymphs were tested for the presence of Borrelia burgdorferi s.l., Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia s.s. DNA from twelve paired areas. Anaplasma phagocytophilum and Babesia s.s. from qPCR-positive tick lysates were identified further to the ecotype and species level, respectively, by DNA sequencing. The infection prevalence of A. phagocytophilum was lower, and infection prevalence of Babesia s.s., identified as Babesia venatorum, was higher in grazed areas. In contrast, infection prevalence with B. burgdorferi s.l. or B. miyamotoi did not differ between grazed and ungrazed areas. As a consequence, conventional cattle grazing in forested areas had no effect on the densities of questing nymphs infected with B. burgdorferi s.l. and B. miyamotoi. Similarly, we found no effect of cattle grazing on the density of infected nymphs with B. venatorum. The marked difference in the densities of questing nymphs infected with A. phagocytophilum could potentially be explained by the presence of a higher density of roe deer (Capreolus capreolus) in ungrazed areas, as the majority of typed A. phagocytophilum from ungrazed areas were the non-zoonotic ecotype II, which is associated with roe deer.     

Borrelia prevalence and species distribution in ticks removed from humans in Germany, 2013–2017

Lyme borreliosis caused by spirochaetes of the Borrelia burgdorferi sensu lato (s.l.) complex is the most common tick-borne disease in Europe. In addition, the relapsing-fever spirochaete Borrelia miyamotoi, which has been associated with febrile illness and meningoencephalitis in immunocompromised persons, is present in Europe. This study investigated Borrelia prevalence and species distribution in ticks removed from humans and sent as diagnostic material to the Institute for Parasitology, University of Veterinary Medicine Hannover, in 2013-2017. A probe-based real-time PCR was carried out and Borrelia-positive samples were subjected to species determination by reverse line blot (RLB), including a B. miyamotoi-specific probe. The overall Borrelia-infection rate as determined by real-time PCR was 20.02 % (510/2547, 95 % CI: 18.48–21.63 %), with annual prevalences ranging from 17.17 % (90/524, 95 % CI: 14.04–20.68 %) in 2014 to 24.12 % (96/398, 95 % CI: 19.99–28.63 %) in 2015. In total, 271/475 (57.1 %) positive samples available for RLB were successfully differentiated. Borrelia afzelii was detected in 30.53 % of cases (145/475, 95 % CI: 26.41–34.89), followed by B. garinii/B. bavariensis (13.26 % [63/475], 95 % CI: 10.34–16.65). Borrelia valaisiana occurred in 5.89 % (28/475, 95 % CI: 3.95–8.41), B. spielmanii in 4.63 % (22/475, 95 % CI: 2.93–6.93), B. burgdorferi sensu stricto (s.s.)/B. carolinensis in 2.32 % (11/475, 95 % CI: 1.16–4.11), B. lusitaniae in 0.63 % (3/475, 95 % CI: 0.13–1.83) and B. bisettiae in 0.42 % (2/475, 95 % CI: 0.05–1.51) of positive ticks. Borrelia kurtenbachii was not detected, while B. miyamotoi was identified in 7.37 % (35/475, 95 % CI: 5.19–10.10) of real-time PCR-positive samples. Sanger sequencing of B. garinii/B. bavariensis-positive ticks revealed that the majority were B. garinii-infections (50/52 successfully amplified samples), while only 2 ticks were infected with B. bavariensis. Furthermore, 6/12 B. burgdorferi s.s./B. carolinensis-positive samples could be differentiated; all of them were identified as B. burgdorferi sensu stricto. Thirty-nine ticks (8.21 %, 95 % CI: 5.90–11.05) were coinfected with two different species. Comparison of the species distribution between ticks removed from humans in 2015 and questing ticks collected in the same year and the same area revealed a significantly higher B. afzelii-prevalence in diagnostic tick samples than in questing ticks, confirming previous observations. The obtained data indicate that Borrelia prevalence fluctuated in the same range as observed in a previous study, analysing the period from 2006 to 2012. Detection of B. miyamotoi in 7.37 % of Borrelia-positive samples points to the fact that clinicians should be aware of this pathogen as a differential diagnosis in cases of febrile illness.     

Use of mammalian museum specimens to test hypotheses about the geographic expansion of Lyme disease in the southeastern United States

Lyme disease, caused primarily in North America by the bacterium Borrelia burgdorferi sensu stricto, is the most frequently reported vector-borne disease in North America and its geographic extent is increasing in all directions from foci in the northeastern and north central United States. Several southeastern states, including Virginia and North Carolina, have experienced large increases in Lyme disease incidence in the past two decades, with the biggest changes in incidence occurring in the western portion of each state. We tested the hypothesis that B. burgdorferi s.s. was present in western Virginia and North Carolina Peromyscus leucopus populations prior to the recent emergence of Lyme disease. Specifically, we examined archived P. leucopus museum specimens, sampled between 1900 and 2000, for B. burgdorferi s.s. DNA. After confirming viability of DNA extracted from ear punch biopsies from P. leucopus study skins collected between 1945 and 2000 in 19 Virginia counties and 17 North Carolina counties, we used qPCR of two species-specific loci to test for the presence of B. burgdorferi s.s. DNA. Ten mice, all collected from the Eastern Shore of Virginia in 1989, tested positive for presence of B. burgdorferi; all of the remaining 344 specimens were B. burgdorferi-negative. Our results suggest that B. burgdorferi s.s was not common in western Virginia or North Carolina prior to the emergence of Lyme disease cases in the past two decades. Rather, the emergence of Lyme disease in this region has likely been driven by the relatively recent expansion of B. burgdorferi s.s. in southward-moving ticks and reservoir hosts in the mountainous counties of these two states.     

A host-restricted viral vector for antigen-specific immunization against Lyme disease pathogen

Newcastle disease virus (NDV) is an avian virus that is attenuated in primates and is a potential vaccine vector for human use. We evaluated NDV as a vector for expressing selected antigens of the Lyme disease pathogen Borrelia burgdorferi. A series of recombinant NDVs were generated that expressed intracellular or extracellular forms of two B. burgdorferi antigens: namely, the basic membrane protein A (BmpA) and the outer surface protein C (OspC). Expression of the intracellular and extracellular forms of these antigens was confirmed in cultured chicken cells. C3H or Balb/C mice that were immunized intranasally with the NDV vectors mounted vigorous serum antibody responses against the NDV vector, but failed to mount a robust response against either the intracellular or extracellular forms of BmpA or OspC. By contrast, a single immunization of hamsters with the NDV vectors via the intranasal, intramuscular, or intraperitoneal route resulted in rapid and rigorous antibody responses against the intracellular or extracellular forms of BmpA and OspC. When groups of hamsters were separately inoculated with various NDV vectors and challenged with B. burgdorferi (10⁸ cells/animal), immunization with vector expressing either intracellular or extracellular BmpA was associated with a significant reduction of the pathogen load in the joints. Taken together, our studies highlighted the importance of NDV as vaccine vector that can be used for simple yet effective immunization of hosts against bacterial infections including Lyme disease.