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Infection caused by neuroinvasive Borrelia often manifests long-term CNS disorders and is difficult to treat as most antibiotics fail to attain an effective concentration within the brain or cannot kill the persister forms of Borrelia (cysts and round bodies). Thus, this study focused on developing antimicrobial cyclic peptides (AMPs) from a combinatorial phage display library that target phosphatidylcholine of the borrelial cell membrane. Isolated cyclic peptides with anti-Borrelia properties were then fused with the CNS homing peptide developed in this study (designated as O-BBB) to facilitate AMP transport across the blood-brain barrier. Among all O-BBB fused AMPs, Bor-18 had half maximal effective concentration (EC50) 0.83 µM when tested against spirochetal Borrelia. Bor-16, Bor-18, and Bor-26 inhibited the cystic form with EC50 0.83 µM, while Bor-11 had EC50 0.41 µM. Within an hour, all four peptides caused a permeability breach in the borrelial cell membrane, causing depolarization of the membrane. Bor peptides did not inhibit eukaryotic cell metabolism or proliferation, nor did they cause erythrocyte lysis. Peptides were stable in serum, could cross the BBB in-vitro, and remained effective against Borrelia. Cyclic AMPs fused with a CNS homing moiety, the Bor peptides, deserve further investigation for their potential use in neuroborreliosis therapy. [ABSTRACT FROM AUTHOR]

Introduction: Lyme disease (LD) is caused by infection with the bacteria Borrelia burgdorferi sensu lato (Bb) through the bite of an infected Ixodes spp. tick. LD has emerged as a public and animal health issue in Canada, with human incidence increasing in part due to the expansion of Ixodes scapularis ticks and their vertebrate hosts. We sought to provide the first comprehensive summary of published tick and animal surveillance literature regarding LD in Canada to describe changes in LD over time. Methods: We conducted a review to identify peer‐reviewed LD‐focused tick, mammal, and bird surveillance articles in three online databases between 1975 and 2023. Data on study characteristics, data collection years, and surveillance methods and findings were extracted. Descriptive statistics were reported. Results: In total, 115 studies were included for review. Results showed an increase in published surveillance literature and changes in study approaches and their provincial distribution over time, coinciding with increased LD cases in Canada. Seventy‐four studies were published after 2014 when Canada's Federal Framework on Lyme Disease Act was introduced, and two‐thirds of these studies focused on tick surveillance only. Overall, 58% of studies involved surveillance in Ontario but increases in all other provinces were observed after 2009. Conclusions: Observed changes in five decades of LD‐related tick and animal surveillance literature helps document the historical rapid spread of Ixodes and Bb across provinces. This can provide lessons for other regions that may transition from emerging to endemic status for LD in the coming years. [ABSTRACT FROM AUTHOR]

Background: Soft ticks (Family: Argasidae) are vectors of relapsing fever Borrelia in the United States and are potential vectors of African swine fever virus, a pathogen that could have a devastating effect on the U.S. swine industry if introduced to the U.S. mainland. Much of the tick-borne disease research in the U.S. focuses on hard ticks, and less is known about the ecology of soft ticks. Some soft tick species found in the southern U.S. have a wide host range and may feed on cattle, swine, native and exotic ungulates, small mammals, reptiles, and humans. Because the feeding habit of most soft tick species involves taking short, repeated blood meals that may include multiple host species, pathogen transmission among hosts is a concern both for human and animal health. Materials and Methods: Sampling was carried out at four locations in south Texas using dry ice traps placed in or near animal burrows and other sheltering cracks and crevasses that may provide refuge for soft ticks. Collected ticks were identified and subsequently screened for Rickettsia and Borrelia species and for host bloodmeal detection using conventional polymerase chain reaction and Sanger sequencing for pathogen and host species identification. Results: In total, 256 ticks of two Ornithodorinae species were screened. Borrelia species were identified in three samples. Bloodmeal detections were made in 22 tick specimens, representing eight vertebrate host species. Conclusions: Results demonstrate that the soft tick species detected herein feed on a range of wildlife hosts in south Texas and are associated with agents of human disease. [ABSTRACT FROM AUTHOR]

The purpose of this study was to assess the prevalence of zoonotic bacteria, including Coxiella burnetii , Bartonella spp. Rickettsia spp. Brucella spp. Borrelia spp. and Ehrlichia spp. among small mammalian in Iran. We examined 618 small mammals collected between 2016 and 2020 from different parts of Iran. We extracted DNA from spleen samples and used quantitative real-time PCR to identify specific genes. We found 559 (90.45%) samples infected with at least one of the analyzed pathogens. Among the studied specimens, 86.08% tested positive for Bartonella spp. 2.42% for Ehrlichia spp. 0.80% for Borrelia spp. 0.64% for C. burnetii , 0.48% for Brucella spp. and 0% for Rickettsia spp. Bartonella krasnovii (25.81%) and Bartonella taylorii (25.81%) were the most prevalent among the Bartonella species. This study identified a rodent infected with Brucella abortus. Among the Borrelia- positive samples, four out of five were identified as Borrelia duttonii. Among the positive cases in the Ehrlichia genus, Ehrlichia canis , Candidatus Ehrlichia shimanensis , and Neoehrlichia mikurensis were identified. Meriones persicus was the most prevalent captured rodent with 315 specimens (51.22%). Our study revealed that a large proportion of the small mammals analyzed were infected with one or more of the targeted pathogens. M. persicus exhibited significant infection rates with C. burnetii , Bartonella spp. Ehrlichia spp. Brucella spp. and Borrelia spp. This suggests that this rodent species could serve as a crucial reservoir for zoonotic pathogens in Iran. [ABSTRACT FROM AUTHOR]

Wild birds may be involved in the transmission of agents of infectious diseases, including zoonoses, a circumstance which raises a number of public and animal health issues. Migratory bird species play a significant role in the introduction of tick-borne pathogens to new geographic areas, contributing to the dissemination of various etiological agents. This preliminary study aimed to assess the occurrence of four potentially zoonotic pathogens (Hepatozoon spp., Borrelia spp., Babesia spp. and Theileria spp.) in the wild birds of Portugal. Blood and tissue samples were taken from 103 birds admitted at wildlife rehabilitation centers. Through the use of conventional PCR, our findings indicate no evidence of the circulation of these pathogens among the studied bird populations in the region. In the One Health context, it is relevant to understand how faraway avian populations play a role in the epidemiology of infectious diseases. Further molecular studies are needed to deepen the knowledge of avian piroplasmosis, borreliosis and hepatozoonosis. [ABSTRACT FROM AUTHOR]

Introduction: In Europe , Borrelia burgdorferi sensu lato (s.l.), the causative agent of Lyme borreliosis is transmitted by the castor bean tick, Ixodes ricinus. In the last decades, global changes affect the spread of ticks and also their bionomics. The aim of this study was summarization of a large dataset obtained during 20 years of research. Methods: The research was carried out in 1999-2019 at 16 localities in Slovakia that were continuously monitored. In total, 17,249 questing I. ricinus ticks were tested for the presence of B. burgdorferi s.l. Results: The total prevalence of infected ticks was 18.8% (3,248/17,249), with 15.1% (1,557/10,302) infected nymphs and 24.3% (1,691/6,947) infected adults. Nine species of B. burgdorferi s.l. were identified. Borrelia afzelii (37.1%), B. garinii / bavariensis (24.7%), and B. valaisiana (15.4%) were the most frequent and were present at all study sites, followed by B. lusitaniae (12.6%), B. burgdorferi sensu stricto (4.1%) and B. spielmanii (1.6%). Borrelia bavariensis was confirmed only in four samples (0.1%), however, detection of this species has been performed only since 2017. Borrelia bissettii and B. kurtenbachii were both recorded in one case. The total prevalence differed significantly among four habitat types (urban, suburban, natural, agricultural). The highest infection prevalence was confirmed in natural habitat (22.0%), the lowest in urban habitat (13.2%). In addition, molecular analysis was carried out on part of the collected ticks previously morphologically identified as I. ricinus. The analysis did not confirm the occurrence of Ixodes inopinatus in Slovakia. Conclusion: Long-term monitoring of the abundance and spread of ticks as well as the prevalence and genetic variability of tick-borne pathogens can reveal the impact of global climatic and socio-economic changes on different habitats, including natural foci of tick-borne pathogens. [ABSTRACT FROM AUTHOR]

Background: Borrelia miyamotoi and Borrelia burgdorferi sensu lato (s.l.) are important zoonotic agents transmitted by Ixodes ricinus ticks, which are widely distributed across Central Europe. Understanding the spatial distribution of these pathogens' prevalence will help identify areas with increased infection risk and facilitate the implementation of effective preventive measures. Methods: We analysed 12,955 I. ricinus ticks collected from 142 towns in the Czech Republic between 2016 and 2018. The ticks were pooled into 2591 groups of five and tested using duplex quantitative polymerase chain reaction (qPCR) for the presence of B. burgdorferi s.l. and B. miyamotoi. For each location, we estimated the overall prevalence of both agents using the EpiTools Epidemiological Calculator for pooled samples and calculated the minimum infection rate (MIR). To assess the potential risk of infection, we combined data on the abundance of nymphs and females with pathogen prevalence at each sampled site. Using a geographic information system (GIS), we mapped the MIR and infection risk of both Borrelia species across all 142 sampled locations and employed a geostatistical method (ordinary kriging) to predict MIR values and infection risk as continuous surfaces across the entire country. Results: We detected B. miyamotoi in 110 localities and B. burgdorferi s.l. in all 142 localities. The estimated prevalence of B. miyamotoi and B. burgdorferi s.l. in the collected ticks was 2.1% (95% confidence interval [CI] 1.8–2.3) and 27.1% (95% CI 26.0–28.3), respectively. For B. miyamotoi, we identified previously unknown, geographically distinct hotspots of MIR up to 8.3%, with MIR slightly higher in females (2.3%) than in males (1.9%) and nymphs (1.8%), though the difference was not statistically significant. In contrast, B. burgdorferi s.l. exhibited ubiquitous presence, with consistently high prevalence nationwide, showing similar MIRs in females (16.2%) and males (16.1%), and slightly lower in nymphs (15.6%). The highest infection risk for B. miyamotoi was 12.4 infected vectors per hour in southeastern Moravia, while the highest risk for B. burgdorferi s.l. reached 78.6 infected vectors per hour in the Bohemian-Moravian Highlands. Conclusions: Borrelia miyamotoi is widespread, forming distinct high-prevalence areas in certain regions. Borrelia burgdorferi s.l. demonstrates consistently high prevalence across most of the country, except for a few localized areas such as southwestern Czechia. Both pathogens exhibit natural nidality, forming regions with elevated prevalence and infection risk. Long-term time-series data are needed to confirm the spatio-temporal stability of these hotspots. [ABSTRACT FROM AUTHOR]

Background: Borrelia infection is caused by Borrelia burgdorferi sensu lato and transmitted by Ixodes ricinus ticks, a common tick-borne infection in Northern Europe. The establishment of Borrelia infection depends on transmission of the spirochetes, as well as the immune response generated in the skin after a bite. Here we aim to investigate the local immune response in the skin after a tick bite and assess the possible direct effects of Borrelia, by applying gene expression analysis of the immune response in skin exposed to Borrelia-infected and non-infected ticks, respectively. Methods: Skin biopsies from the study participants were taken 7–10 days after the tick-bite. The ticks and skin biopsies were analysed by real-time PCR for Borrelia spp. and other tick-borne pathogens. Dermal transcriptome profiles derived from RNA sequencing with focus on immune system regulation were created. In addition, we performed enrichment analysis of dermal transcriptome profiles with focus on immune system regulation. Results: Skin biopsies exposed to a Borrelia-positive tick induced an overall higher expression of immune-related genes. Cytokines involved in the regulation of T-cell and macrophage activation, pro-inflammatory regulators and Toll-like receptor 2, 3 and 7 involved in pathogen recognition were upregulated in skin exposed to Borrelia, although Borrelia DNA was not detected in the biopsies. Conclusion: The evidence of upregulation of genes in Borrelia exposed skin suggests an influence on the immune system of ticks and spirochetes. Characterization of Borrelia-associated gene expression signatures in the skin could contribute to future diagnostics and increase our understanding of the development of various manifestations of Borrelia infection. [ABSTRACT FROM AUTHOR]

Ticks are pivotal in transmitting a variety of pathogens that affect both humans and animals. These pathogens often occur in guilds, groups of species that exploit similar resources in similar ways. Although the composition of tick‐borne pathogen (TBP) guilds is well‐documented, the interactions among pathogens within these guilds remain poorly understood. We hypothesized that abiotic and biotic factors significantly influence the patterns of occurrence and interactions among pathogens within these guilds. To investigate this, we analyzed microfluidic‐based high‐throughput data on microorganisms from 166 Hyalomma excavatum ticks (94 male and 72 female) collected across different seasons from cattle in the central Algerian steppe using network analysis to uncover complex pathogen–pathogen interaction patterns. We found that female ticks had a higher infection rate (63.9%) with common pathogens such as Rickettsia slovaca (26.4%), unclassified Apicomplexa (22.2%), and Borrelia afzelii (19.4%). Male ticks showed a 56.4% infection rate, with Rickettsia (31.1%) and R. slovaca (16%) being the most prevalent. Notable pathogen–pathogen interactions within guilds were identified, with positive associations such as between R. slovaca and Rickettsia conorii in males, and B. afzelii and Borrelia spielmanii in females, indicating cooperative interactions. Conversely, negative associations, such as between Anaplasma phagocytophilum and Francisella tularensis, suggested competitive exclusion. The observed variation in interaction patterns under different conditions indicates that ecological determinants, both biotic and abiotic, influence pathogen association dynamics within guilds. These findings have significant implications for understanding disease transmission and developing control strategies. [ABSTRACT FROM AUTHOR]

ABSTRACT Introduction Methods Results Conclusions Ixodes ricinus, the castor bean tick, is the most prevalent tick species in Europe. It favours habitats such as shrubs, deciduous and mixed forests, but can also be found in urban environments. Due to its high vector competence, it is of enormous veterinary as well as medical importance, transmitting tick‐borne encephalitis (TBE) virus, Borrelia burgdorferi s. l., the causative agent of lyme borreliosis, Rickettsia spp. and Anaplasma phagocytophilum amongst many other pathogens. In Kosovo, I. ricinus is the predominant species and a few studies, mostly based on human and animal seroprevalences, indicate the circulation of tick‐borne pathogens. However, data on pathogens in I. ricinus are scarce in Kosovo, particularly in urban settings. This study aimed to provide first insights into the circulation of tic‐kborne pathogens in I. ricinus from urban and peri‐urban areas in Kosovo.Urban and peri‐urban areas were sampled by flagging 150 m transects. In total, 197 ticks were morphologicaly identified as I. ricinus, and consequently DNA and RNA were isolated. All individuals were screened for the presence of tick‐borne pathogens by using reverse line blotting (RLB) hybridisation.DNA of nine different pathogens from four genera including Borrelia spp., Rickettsia spp., Anaplasma spp. and Babesia spp. was detected in 60 (33.5%) specimens. The most frequently detected pathogens were Rickettsia spp. (16.2%), followed by Borrelia spp. (11.7%). Altogether, 54 single infections, 11 double infections and 1 triple infection were observed.We provide first data on genotyping of B. burgdorferi sensu lato as well as the detection of Anaplasma, Babesia and Rickettsia from I. ricinus in this country. The data underline that particularly recreational (peri‐)urban areas could facilitate the spillover of zoonotic tick‐borne pathogens to humans in Kosovo and provide baseline data for future surveys. [ABSTRACT FROM AUTHOR]

Tick-borne diseases are a public health problem and a significant burden on the livestock industry. The seasonal abundance of ticks and tick-borne pathogens strongly correlates with the prevalence of these diseases. To investigate the seasonal variation in ticks and tick-borne pathogens, ticks were collected from Gangwon State, Korea, and the tick-borne pathogens Borrelia, Anaplasma, Babesia, and Theileria were examined. In total, 14,748 ticks were collected, comprising ticks from two genera and three species: Haemaphysalis longicornis, Haemaphysalis flava, and Ixodes nipponensis, with H. longicornis being the predominant species. Of 7445 ticks (455 pools) examined for pathogens, Theileria was detected in 61 pools, whereas Borrelia and Anaplasma were observed in 17 pools. H. longicornis nymphs and adults were collected beginning in April, with nymph numbers peaking in May and June and adult ticks peaking in June and July. In contrast, the larvae were collected in May and peaked in September. Tick-borne pathogens were detected in April, peaking in July and September. Borrelia, the causative agent of Lyme disease, exhibits a temporal association between its detection in ticks and its occurrence in humans. In conclusion, tick-borne diseases seem to be closely linked not only to changes in tick numbers throughout the seasons but also to the seasonal variations of the pathogens within them. [ABSTRACT FROM AUTHOR]

Tick-borne pathogens are growing in importance for human and veterinary research worldwide. We developed, optimized, and validated a reliable quantitative PCR (qPCR; real-time PCR) assay to assess Borrelia burgdorferi infection by targeting two B. burgdorferi genes, ospA and flaB. When assessing previously tested tick samples, its performance surpassed the nested PCR in efficiency, sensitivity, and specificity. Since the detection of Borrelia is more difficult in mammalian samples, the qPCR assay was also assessed using wildlife tissues. For wildlife samples, the sensitivity and specificity of ospA primers, with the incorporation of a pre-amplification step, was equivalent or superior to the nested PCR. For human samples, no primer set was successful with human tissue without culture, but we detected Borrelia with ospA and flaB primers in 50% of the Lyme culture samples, corresponding to 60% of the participants with a Lyme disease diagnosis or suspicion. The specificity of amplification was confirmed by Sanger sequencing. The healthy participant culture samples were negative. This PCR-based direct detection assay performs well for the detection of Borrelia in different biological samples. Advancements in detection methods lead to a better surveillance of Borrelia in vectors and hosts, and, ultimately, enhance human and animal health. [ABSTRACT FROM AUTHOR]

Lyme arthritis, one of the possible late manifestations of Lyme borreliosis, predominantly affects the supporting joints and in adults most often occurs in the form of monoarthritis of the knee. Early diagnosis is based on clinical findings and serology. PCR detection of Borrelia in synovial fluid has become an integral part of the laboratory testing algorithm. The clinical presentation and inflammatory markers in Lyme arthritis can resemble septic arthritis. Determining the levels of alpha-defensins (human neutrophil peptide (HNP 1–3)) in synovial fluid by liquid chromatography is a highly sensitive method revealing the presence of inflammatory process. Between 2020 and 2022, we examined eleven patients with Lyme arthritis of the knee. We measured levels of HNP 1–3 from synovial fluid by HPLC in patients, and we compared it with the corresponding C-reactive protein (CRP) levels in paired serum samples. In patients diagnosed with Lyme arthritis, HNP 1–3 levels in synovial fluid ranged from 2.5 to 261 mg/L, with a median of 46.5 mg/L. Average serum CRP was 43 mg/L. The results show that elevated HNP 1–3 can be consistent with not only septic arthritis or systemic disease, but also with Lyme arthritis, especially in patients with negative culture and 16S PCR from synovial fluid. Final diagnosis must be verified by examination for anti-Borrelia antibodies from serum and synovial fluid. The aim of this work is to introduce an HPLC method for the determination of alpha-defensins as one of the possible diagnostic markers. [ABSTRACT FROM AUTHOR]

Abstract 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. Graphical abstract

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. [ABSTRACT FROM AUTHOR]

The genus Borrelia encompasses a diverse group of spirochetes transmitted primarily by ticks, with Borrelia burgdorferi causing Lyme disease, which is prevalent in North America and Europe. Borrelia's structural adaptations and ability to persist in diverse host tissues underscore its pathogenic potential. Beyond traditional infectious responses, Borrelia engages in complex interactions with the host immune system, contributing to autoimmune mechanisms such as molecular mimicry and persistent infections. This intricate interplay manifests in symptoms resembling various autoimmune diseases, including systemic lupus erythematosus, dermatomyositis, local scleroderma, and systemic sclerosis. However, these associations lack a precise explanation, emphasizing the need for further investigation. The cases of misdiagnosis between Lyme borreliosis and autoimmune diseases highlight the critical importance of accurate diagnostics and adherence to guidelines. Understanding Borrelia's impact on immune responses is pivotal for advancing diagnostics and targeted therapeutic interventions in Lyme borreliosis and its potential autoimmune implications. [ABSTRACT FROM AUTHOR]

Lyme disease is a vector-borne illness caused by spirochetes belonging to the Borrelia burgdorferi species group. These bacteria employ several mechanisms to survive within the vertebrate host, including evasion of the complement system. In this study, we examine the protection against human serum killing by the binding of host complement regulators C4b-binding protein (C4BP) and factor H (FH) to the bacterial surface of B. burgdorferi. Via serum depletion of isolated complement regulators, we found that the absence of C4BP did not alter the survival of B. burgdorferi strain B31; however, the removal of FH increased the sensitivity of this strain to human serum as previously described. The B. garinii seabird-isolated strain Far04, on the other hand, did not bind any complement regulators of human origin and was serum-sensitive, indicating its special host species specificity. [ABSTRACT FROM AUTHOR]

Lyme borreliosis (LB) is not notifiable in many European countries, and the patchwork of surveillance strategies in Europe perpetuates knowledge gaps. In the Netherlands, LB incidence has been estimated from recurring general practitioner surveys since the 1990s. To complement the incidence data, this study aimed to estimate the prevalence of antibodies against Borrelia burgdorferi sensu lato in the general population of the Netherlands in 1995/1996, identify risk factors for seropositivity, and compare these findings to data from 2016/2017 to identify temporal trends. Sera from participants (n = 8041, aged 0–80 years) in a cross-sectional nationwide surveillance study were assessed for the presence of antibodies against B. burgdorferi s.l., using a screening ELISA and immunoblot confirmation. Risk factors associated with seropositivity were evaluated using multivariable analysis. A significant difference in weighted seroprevalence was observed between 1995/1996 (2.8%) and 2016/2017 (4.3%). In both cohorts, the seroprevalence was significantly higher among men than among women, and increased with age and tick bite frequency. The upward trend in age-specific seropositivity in individuals over 50 was steeper in 2016/2017 than in 1995/1996, possibly due to improved fitness among contemporary elderly, allowing increased outdoor activities. This study highlights significant trends in the seroprevalence of B. burgdorferi s.l. antibodies in the general population of the Netherlands over 20 years. The doubling of seroprevalence underscores the increasing burden of LB, and the importance of continued surveillance. Targeted interventions, particularly for elderly populations, may help raise awareness to the risks of tick bites and reduce the growing disease burden and societal costs associated with LB. [ABSTRACT FROM AUTHOR]

VMP-like sequence (vls) antigenic variation systems are present in every Lyme disease Borrelia strain with complete genome sequences. The linear plasmidencoded vls system consists of a single expression site (vlsE) and contiguous array(s) of silent cassettes that have ~90% identity with the central cassette region of the cognate vlsE gene; antigenic variation occurs through random, segmental, and unidirectional recombination of vls silent cassette sequences into the vlsE expression site. Automated annotation programs do not accurately recognize vls silent cassette sequences, so these regions are not correctly annotated in most genomic sequences. In this study, the vls sequences were re-analyzed in the genomic sequences of 31 available Lyme disease Borrelia and one relapsing fever Borrelia organisms, and this information was utilized to systematically compare the vls systems in different species and strains. In general, the results confirm the conservation of the overall architecture of the vls system, such as the head-to-head arrangement of vlsE and a contiguous series of vlsS silent cassette sequences and presence of inverted repeat sequences between the two regions. However, the data also provide evidence for the divergence of the vls silent cassette arrays through point mutations, short indels, duplication events, and rearrangements. The probable occurrence of convergent evolution toward a vls system-like locus is exemplified by Borrelia turcica, a variable large protein (Vlp) expressing organism that is a member of the relapsing fever Borrelia group. [ABSTRACT FROM AUTHOR]

Although the importance of the microbiome in the context of tick biology and vector competence has recently come into a broader research focus, the field is still in its infancy and the complex ecological interactions between the tick residential bacteria and pathogens are obscure. Here, we show that an environmentally acquired gut bacterium has the potential to impair Borrelia afzelii colonization within the tick vector through a secreted metalloprotease. Oral introduction of either Bacillus cereus LTG-1 isolate or its purified enhancin (BcEnhancin) protein significantly reduces B. afzelii burden in the guts of Ixodes ricinus ticks. This effect is attributed to the ability of BcEnhancin to degrade a glycan-rich peritrophic matrix (PM), which is a gut protective barrier essential for Borrelia survival. Our study highlights the importance of the gut microbiome in determining tick vector competence and provides a deeper mechanistic insight into the complex network of interactions between Borrelia, the tick, and the tick microbiome. [ABSTRACT FROM AUTHOR]

The Borrelia burgdorferi sensu lato (s.l.) complex includes a group of spirochete bacteria that are involved in transmission cycles with vertebrates and the ticks associated with them. Rodents play an essential role in the ecoepidemiology of Borrelia, acting as reservoirs for the bacteria and hosts for ticks. To identify potential reservoir vertebrate hosts of Borrelia spirochetes in Argentina, we conducted molecular analyses on tissues obtained from a population of Sigmodontinae rodents inhabiting the Delta and Parana Islands ecoregion. We report the presence of B. burgdorferi s.l. complex genotypes infecting Akodon azarae and Oxymycterus rufus in this ecoregion. Phylogenetic analysis revealed at least three flaB haplotypes related to ‘Candidatus Borrelia paulista’ and Borrelia sp. strain Pampa from Brazil and to several Borrelia haplotypes from Uruguay. Additionally, we identified different alleles of the plasmid‐borne gene ospC in B. burgdorferi s.l. infecting A. azarae and Ox. rufus. The ospC allele present in Borrelia from Ox. rufus was also detected in Borrelia from Oligoryzomys mattogrossae, indicating the ability of Borrelia harbouring this ospC allele to infect multiple Sigmodontinae species, whereas the ospC allele from Borrelia infecting A. azarae represents a novel variant. Further research is needed to determine host specificity of ospC alleles present in South America. [ABSTRACT FROM AUTHOR]

The article addressed the issue of diagnosing Lyme disease in dogs. The study aimed to analyse the effectiveness of contemporary diagnostic methods for Lyme disease in dog populations. A review of the literature was conducted, examining studies that explored various diagnostic approaches. Based on the literature analysis, a classification of diagnostic methods was developed. This classification encompassed general diagnostic methods - including molecular, serological, and bacterioscopic techniques - as well as the application of nanotechnologies for diagnosing the causative agent of this disease (Borrelia burgdorferi) in Ixodes ticks (Ixodes ricinus, I. hexagonus, and I. persulcatus). The general diagnostic methods for Lyme disease in dogs also include several advanced high-precision techniques. These molecular methods comprise polymerase chain reaction, quantitative real-time polymerase chain reaction, and polymerase chain reaction dissociation. Serological methods include enzyme-linked immunosorbent assay, immunofluorescent assay, and immunoblotting. Bacterioscopic methods involve cultural techniques, immunohistochemical analysis, and microfluidics. Among the nanotechnologies, complete analysis microsystems and electrochemical methods were identified. Tick diagnosis for Lyme disease includes techniques aimed at detecting the presence of pathogens, specifically Borrelia species, within the tick itself. The analysis of diagnostic methods provided their characteristics and highlighted promising approaches for identifying Lyme disease in dogs. Among these, enzyme-linked immunosorbent assay and immunofluorescent assay yielded the best results due to their cost-effectiveness and rapid output, requiring minimal pathological material obtained at various stages of the disease. It was established that one of the most promising diagnostic tools for Lyme disease in dogs is the use of biomarkers. Notable examples include protein markers of inflammation, cytokines, chemokines, and genetic biomarkers. The findings from the literature analysis of diagnostic methods for Lyme disease in dogs will be valuable for veterinary practitioners involved in the treatment of vector borne diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: Ixodes ricinus, the most important tick vector in Europe, is responsible for the spreading of medically important pathogens. This study aimed to detect Borrelia spp. and Rickettsia spp. in I. ricinus ticks collected in urban and protected areas both in Poland and the Czech Republic. Ixodes ricinus ticks collected in Poland compared to ticks from the Czech Republic were more infected with Borrelia spp., and ticks collected in protected areas were more infected with Borrelia spp. than ticks collected in urban areas. Rickettsia spp. prevalence in I. ricinus was comparable in both Poland and the Czech Republic; however, regional differences were observed. The regional differences in Borrelia spp. and Rickettsia spp. prevalence in I. ricinus indicate the need for adaptation of public health surveillance strategies in each region. (1) Background: Ixodes ricinus is responsible for the spreading of medically important pathogens. Monitoring the level of tick infection in various areas is essential for determining the potential tick-born risk. This study aimed to detect Borrelia spp. and Rickettsia spp. in I. ricinus ticks collected in urban and protected areas both in Poland and the Czech Republic. (2) Methods: Ticks were collected by flagging in the years 2016–2017. Borrelia spp. was detected using nested PCR targeting the flaB gene and Rickettsia spp. using nested PCR targeting gltA. (3) Results: In total, DNA of Borrelia spp. was detected in 25.9% of samples. Ticks collected in Poland were more infected compared to the Czech Republic and ticks collected in protected areas were more infected with Borrelia spp. than ticks collected in urban areas. The RFLP analysis showed the occurrence of B. afzelii and B. garinii in both countries, and additionally B. valaisiana, B. burgdorferi s.s., and B. miyamotoi in Poland. Rickettsia spp. was detected in 17.4% of I. ricinus, with comparable infection level in both countries; however, regional differences were observed. (4) Conclusion: The regional differences in Borrelia spp. and Rickettsia spp. prevalence in I. ricinus indicate the complexity of factors influencing the level of infection and underline the need for adaptation public health surveillance strategies in each region. [ABSTRACT FROM AUTHOR]

Soft ticks in the genus Ornithodoros occur throughout the Mojave Desert in southern Nevada, southeastern California, and parts of southwestern Utah and northwestern Arizona, USA, and are frequently observed parasitizing Mojave desert tortoises (Gopherus agassizii). However, limited research exists examining the relationship between ticks and desert tortoises. Mojave desert tortoises are listed as threatened by the US Fish and Wildlife Service, and as such, their populations are monitored and individual tortoise health is routinely assessed. These health assessments document the presence and abundance of ticks present on tortoises, but detailed examination of the relationship between ticks and tortoise health has been lacking. This study analyzed the relationship between tick presence and desert tortoise health assessments as a function of season, location, age (adult vs. juvenile), foraging behavior, evidence of clinical signs of disease, body condition score, and sex. Our results indicate that more ticks were found on tortoises in the summer than in any other season. Ticks were observed more frequently on captive tortoises versus wild tortoises, and more ticks were likely to be present on adult tortoises than on juveniles. Ticks were also more likely to be observed on tortoises that lacked evidence of foraging and on tortoises with observed clinical signs of disease. These findings provide valuable insights into the biology of ticks in relation to tortoises that may be useful for management of both captive and free-living threatened tortoise populations where ticks are detected. Our study also may improve understanding of potential tick-borne disease dynamics in the Mojave desert tortoise habitat, including Borrelia sp. carried by Ornithodoros ticks, which cause tick- borne relapsing fever in people. [ABSTRACT FROM AUTHOR]

Compared to classic Lyme disease (LD), Baggio–Yoshinari syndrome (BYS) has the following distinctive characteristics: it is transmitted in the Amazon area and Northeast, Central-West, Southeast, and South regions of Brazil by hard ticks, notably Amblyomma cajannense or Rhipicefalus sp. The absence of Ixodes sp. ticks in areas at risk of BYS in Brazil is probably the main reason for the disease's differences from LD in the United States, Europe, and Asia. Biodiversity and climate probably favor the formation of atypical pleomorphic Borrelias, which have not yet been cultivated or isolated. Clinically, the first manifestation of BYS is the erythema migrans as in the classic forms of Lyme disease, but BYS is distinguished from LD by its prolonged clinical evolution, with a high frequency of relapses and the appearance of autoimmune manifestations. Prevalent symptoms are headache and erythema nodosum. Five clinical cases of BYS in patients who contracted the disease in the Brazilian Amazon rainforest are described here. This syndrome should be considered among differential diagnoses in patients bitten by ticks in Brazil who present with erythema migrans and/or headache. It is important to pursue an early diagnosis because symptoms respond well to antibiotics in the early stages; if treatment is started late, a chronic course with articular and neurological sequelae can be detected. [ABSTRACT FROM AUTHOR]

The European red squirrel (Sciurus vulgaris) is a common host for Ixodes ricinus ticks in urban and rural habitats, however, studies on ticks and tick-borne pathogens (TBPs) of squirrels have not been conducted in Poland yet. Thus, the aims of the current study were to assess and compare the prevalence and abundance of ticks on red squirrels trapped at two sites in the Warsaw area (in an urban forest reserve and an urban park) and using molecular tools, to assess the genetic diversity of three pathogens (Borrelia burgdorferi sensu lato, Rickettsia and Babesia spp.) in I. ricinus ticks collected from squirrels. For the detection of Rickettsia spp. a 750 bp long fragment of the citrate synthase gltA gene was amplified; for B. burgdorferi s.l. 132f/905r and 220f/824r primers were used to amplify the bacterial flaB gene fragments (774 and 605 bp, respectively) and for Babesia spp., a 550 bpfragment of 18S rRNA gene was amplified. In total, 91 red squirrels were examined for ticks. There were differences in tick prevalence and mean abundance of infestation in squirrels from the urban forest reserve and urban park. Three species of B. burgdorferi s.l., Rickettsia spp., and Babesia microti were detected in ticks removed from the squirrels. Our results broaden knowledge of S. vulgaris as an important host for immature I. ricinus stages and support the hypothesis that red squirrels act as a reservoir of B. burgdorferi. Moreover, we conclude that red squirrels may also play a role in facilitating the circulation of other pathogens causing serious risk of tick-borne diseases in natural and urban areas. [ABSTRACT FROM AUTHOR]

Tick-borne pathogen emergence is dependent on the abundance and distribution of competent hosts in the environment. Ixodes scapularis ticks are generalist feeders, and their pathogen infection prevalence depends on their relative feeding on local competent and non-competent hosts. The ability to determine what host a larval life stage tick fed on can help predict infection prevalence, emergence, and spread of certain tick-borne pathogens and the risks posed to public health. Here, we use a newly developed genomic target-based technique to detect the source of larval bloodmeals by sampling questing nymphs from Block Island, RI, a small island with a depauperate mammalian community. We used previously designed specific assays to target all known hosts on this island and analyzed ticks for four human pathogenic tick-borne pathogens. We determined the highest proportion of larvae fed on avian species (42.34%), whitefooted mice (36.94%), and white-tailed deer (20.72%) and occasionally fed on feral cats, rats, and voles, which are in low abundance on Block Island. Additionally, larvae that had fed on white-footed mice were significantly more likely to be infected with Borrelia burgdorferi and Babesia microti, while larvae that had fed on white-footed mice or white-tailed deer were significantly more likely to be infected with, respectively, mouseand deer-associated genotypes of Anaplasma phagocytophilum. The ability to detect a nymph's larval host allows for a better understanding of tick feeding behavior, host distribution, pathogen prevalence, and zoonotic risks to humans, which can contribute to better tick management strategies. [ABSTRACT FROM AUTHOR]

Five chromosomally encoded proteins, BB0108, BB0126, BB0298, BB0323, and BB0689, from Borrelia burgdorferi sensu lato (s.l.), were obtained in three variants each, representing the most common genospecies found in Europe (Borrelia afzelii, Borrelia burgdorferi sensu stricto (s.s.), and Borrelia garinii). The reactivity of these recombinant proteins with the IgM and IgG antibodies present in human serum was assessed using Western blot (WB) and the ELISA. In IgG-WB, the proteins exhibited varying reactivity, peaking at approximately 40–50% for BB0108 and BB0689. However, none of these proteins were recognized by specific antibodies in the IgM-WB. The sensitivity of IgG-ELISA based on three variants of BB0108 and BB0323 ranged from 71% to 82% and from 62% to 72%, respectively. Conversely, the specificity of both tested proteins was consistently above 82%. Tests utilizing single variants of BB0323 did not yield any diagnostic value in detecting IgM antibodies. However, BB0108 demonstrated recognition by antibodies present in 52% to 63% of the tested sera. These antigens appear advantageous due to the consistent reactivity observed across their variants. This observation suggests that appropriate selection of antigens conserved within B. burgdorferi s.l. could offer a solution to the issue of variable sensitivity encountered in serodiagnostic tests across Europe. [ABSTRACT FROM AUTHOR]

This study reveals a significant presence of ticks and tick-borne pathogens in urban recreational areas of Tallinn, Estonia. During the period of May–June 2018, 815 Ixodes ticks were collected from an area of 11,200 m2 using the flagging method. Tick density reached up to 18.8 ticks per 100 m2, indicating a high concentration of ticks in these urban green spaces. Pathogen analysis demonstrated that 34% of the collected ticks were infected with at least one pathogen. Specifically, Borrelia burgdorferi s.l., the causative agent of Lyme borreliosis, was detected in 17.4% of the ticks; Rickettsia spp. was detected in 13.5%; Neoehrlichia mikurensis was detected in 5.5%; Borrelia miyamotoi was detected in 2.6%; and Anaplasma phagocytophilum and tick-borne encephalitis virus were detected in 0.5% each. These findings indicate that the prevalence and abundance of ticks and tick-borne pathogens in these urban environments are comparable to or even exceed those observed in natural endemic areas. Given the increasing incidence of Lyme borreliosis in Central and Northern Europe, the risk of tick bites and subsequent infection in urban recreational sites should not be underestimated. Public health measures, including enhanced awareness and precautionary information, are essential to mitigate the risk of tick-borne diseases in these urban settings. [ABSTRACT FROM AUTHOR]

Borrelia burgdorferi sensu lato complex comprises 20 species, from which B. afzelii, B. garinii, B. burgdorferi sensu stricto, B. bavariensis, and B. spielmanii are directly associated with Lyme borreliosis, while B. bissettiae, B. lusitaniae, and B. valaisiana were detected in individual cases. Their main vector in Europe is the hard tick species Ixodes ricinus. To date, two species, B. spielmanii and B. lusitaniae, have been molecularly detected in Bosnia and Herzegovina. To test for the presence of other Borrelia species, we performed nested PCR targeting intergenic region rrf (5S) – rrl (23S) on DNA isolates from 49 ticks collected from vegetation by flagging method and 43 removed from humans in The Center for Emergency Medical Assistance of the Sarajevo Canton and The Healthcare Centers of the Sarajevo Canton. Borrelia species were detected by one-directional Sanger sequencing of the amplified region using the same forward primer as in PCR. Out of six Borrelia species detected in the present study, this is the first record of B. afzelii, B. garinii, B. burgdorferi s.s. B. bavariensis, and B. valaisiana in Bosnia and Herzegovina. [ABSTRACT FROM AUTHOR]

ABSTRACT The bacterium responsible for Lyme disease, Borrelia burgdorferi, accumulates high levels of manganese without iron and possesses a polyploid genome, characteristics suggesting potential extreme resistance to radiation. Contrary to expectations, we report that wild-type B. burgdorferi B31 cells are radiosensitive, with a gamma-radiation survival limit for 106 wild-type cells of

ABSTRACT Despite its importance in pathogenesis, the hematogenous dissemination pathway of Borrelia burgdorferi is still largely uncharacterized. To probe the molecular details of transendothelial migration more easily, we studied this process using cultured primary or telomerase-immortalized human microvascular endothelial cells in a medium that maintains both the human cells and the spirochetes. In B. burgdorferi-infected monolayers, we observed ~55% of wild-type spirochetes crossing the monolayer. Microscopic characterization revealed entrance points across the cellular surface rather than at cellular junctions, supporting a transcellular route. In support of this pathway, locking the endothelial junctions using a vascular endothelial protein tyrosine phosphatase (VE-PTP) inhibitor did not reduce transendothelial migration. We also used inhibitors to block the most common endocytic pathways to elucidate effectors that might be involved in B. burgdorferi uptake and/or transmigration. Directly inhibiting Cdc42 reduced spirochete transmigration by impeding internalization. However, blocking Rac1 alone dramatically reduced transmigration by ~84% and resulted in a concomitant doubling in spirochete accumulation in the cell. Our combined results support that B. burgdorferi internalization is an intermediate step in the transendothelial migration process, which requires both Cdc42 and Rac1; Cdc42 is needed for spirochete internalization, while Rac1 is required for cellular egress. These are the first two host proteins implicated in B. burgdorferi transmigration across endothelial cells.IMPORTANCELyme borreliosis is caused by Borrelia burgdorferi and related bacteria. It is the most common tick-transmitted illness in the Northern Hemisphere. The ability of this pathogen to spread to a wide variety of locations results in a diverse set of clinical manifestations, yet little is known regarding vascular escape of the spirochete, an important pathway for dissemination. Our current work has studied the traversal of B. burgdorferi across a monolayer of microvascular endothelial cells grown using a new culture system. We show that this occurs by passage of the spirochetes directly through cells rather than at cellular junctions and that internalization of B. burgdorferi is an intermediate step in transmigration. We also identify the first two host proteins, Cdc42 and Rac1, that are used by the spirochetes to promote traversal of the cellular monolayer. Our new experimental system also provides a new avenue for further studies of this important process.

IntroductionIn Europe, Borrelia burgdorferi sensu lato (s.l.), the causative agent of Lyme borreliosis is transmitted by the castor bean tick, Ixodes ricinus. In the last decades, global changes affect the spread of ticks and also their bionomics. The aim of this study was summarization of a large dataset obtained during 20 years of research.MethodsThe research was carried out in 1999-2019 at 16 localities in Slovakia that were continuously monitored. In total, 17,249 questing I. ricinus ticks were tested for the presence of B. burgdorferi s.l. ResultsThe total prevalence of infected ticks was 18.8% (3,248/17,249), with 15.1% (1,557/10,302) infected nymphs and 24.3% (1,691/6,947) infected adults. Nine species of B. burgdorferi s.l. were identified. Borrelia afzelii (37.1%), B. garinii/bavariensis (24.7%), and B. valaisiana (15.4%) were the most frequent and were present at all study sites, followed by B. lusitaniae (12.6%), B. burgdorferi sensu stricto (4.1%) and B. spielmanii (1.6%). Borrelia bavariensis was confirmed only in four samples (0.1%), however, detection of this species has been performed only since 2017. Borrelia bissettii and B. kurtenbachii were both recorded in one case. The total prevalence differed significantly among four habitat types (urban, suburban, natural, agricultural). The highest infection prevalence was confirmed in natural habitat (22.0%), the lowest in urban habitat (13.2%). In addition, molecular analysis was carried out on part of the collected ticks previously morphologically identified as I. ricinus. The analysis did not confirm the occurrence of Ixodes inopinatus in Slovakia.ConclusionLong-term monitoring of the abundance and spread of ticks as well as the prevalence and genetic variability of tick-borne pathogens can reveal the impact of global climatic and socio-economic changes on different habitats, including natural foci of tick-borne pathogens.

Persistent symptoms after an infection have been described for a number of infectious diseases, including Lyme disease. Studies have confirmed a moderate but consistent increase in the prevalence of such symptoms after Lyme disease, though the risk increase varies dependent on study design and the definition of persistent symptoms. Various possible predictors have been proposed, including a dysregulation of the immune system, metabolic changes, increased sensitization to pain signals, cognitive-behavioral factors, or—controversially—the persistence of the causative Borrelia bacteria or remnants thereof. Research on the precise roles of any of these factors is still ongoing. The lack of biological underpinning also makes it difficult to assess with certainty which patients' (generally nonspecific) persistent symptoms are etiologically related to the previous Lyme disease episode and which are not, particularly as these symptoms occur in the general population relatively frequently. The diagnostic criteria for posttreatment Lyme disease syndrome have shown their usefulness in both clinical and research settings but leave out a number of patients whose symptoms may fall just outside said criteria. Though the relationship between these symptoms and the previous Lyme disease episode may be very uncertain, we would argue that a uniform description and classification of these patients will aid in future research and patient management, regardless of the eventual underlying cause. Thus, we argue for an inclusive classification system for all persistent symptoms attributed to Lyme disease in order to promote validation of patient experiences and perspectives, while also maintaining scientific nuance regarding the very uncertain etiology of these patients' symptoms. [ABSTRACT FROM AUTHOR]

Lyme disease is caused by the spirochete, Borrelia burgdorferi, which is transmitted by Ixodes spp ticks. The rise in Lyme disease cases since its discovery in the 1970s has reinforced the need for a vaccine. A vaccine based on B burgdorferi outer surface protein A (OspA) was approved by the Food and Drug Administration (FDA) several decades ago, but was pulled from the market a few years later, reportedly due to poor sales, despite multiple organizations concluding that it was safe and effective. Newer OspA-based vaccines are being developed and are likely to be available in the coming years. More recently, there has been a push to develop vaccines that target the tick vector instead of the pathogen to inhibit tick feeding and thus prevent transmission of tick-borne pathogens to humans and wildlife reservoirs. This review outlines the history of Lyme disease vaccines and this movement to anti-tick vaccine approaches. [ABSTRACT FROM AUTHOR]

Borrelia miyamotoi, transmitted by Ixodes spp. ticks, was recognized as an agent of hard tick relapsing fever in the United States in 2013. Nine state health departments in the Northeast and Midwest have conducted public health surveillance for this emerging condition by using a shared, working surveillance case definition. During 2013-2019, a total of 300 cases were identified through surveillance; 166 (55%) were classified as confirmed and 134 (45%) as possible. Median age of case-patients was 52 years (range 1-86 years); 52% were male. Most cases (70%) occurred during June-September, with a peak in August. Fever and headache were common symptoms; 28% of case-patients reported recurring fevers, 55% had arthralgia, and 16% had a rash. Thirteen percent of patients were hospitalized, and no deaths were reported. Ongoing surveillance will improve understanding of the incidence and clinical severity of this emerging disease. [ABSTRACT FROM AUTHOR]

Introduction: Lyme disease (LD), caused by the spirochete Borrelia burgdorferi, is the most common vector‐borne disease in the United States. Although most surveillance‐reported cases are in people who are White, data suggest worse outcomes among people from racial and ethnic minority groups. Methods: We conducted a systematic literature review to describe racial disparities in LD. We described the epidemiology of LD by race and ethnicity, including clinical presentation at diagnosis, and summarised the literature on knowledge, attitudes and practices related to LD and ticks by race and ethnicity. Results: Overall, the incidence and prevalence of LD were 1.2–3.5 times higher in White persons than in persons who identified as Asian or Pacific Islander and 4.5–6.3 times higher in White persons than in persons who identified as Black. Across multiple studies, people from racial and ethnic minority groups were more likely than White people to have disseminated manifestations of LD, including neurological manifestations and arthritis, and less likely to have erythema migrans. People from racial and ethnic minority groups were also more likely to report disease onset in the fall and less likely to report disease onset in the summer. Possible reasons for these disparities include lack of recognition of the disease in people with darker skin tones, lack of knowledge of disease risk for some groups and differences in exposure risk. Conclusions: Taken together, these results reinforce that all people residing in high‐incidence areas are at risk of LD, regardless of race or ethnicity. Future prevention measures should be broadly targeted to reach all at‐risk populations. [ABSTRACT FROM AUTHOR]

Due to the extensive use of green urban areas as recreation places, city residents are exposed to tick-borne pathogens. The objectives of our study were (i) to determine the occurrence of ticks in urban green areas, focussing on areas used by humans such as parks, schools and kindergartens, and urban forests, and (ii) to assess the prevalence of Borrelia infections in ticks in Zielona Góra, a medium-sized city in western Poland. A total of 161 ticks representing the two species Ixodes ricinus (34 males, 51 females, 30 nymphs) and Dermacentor reticulatus (20 males, 26 females) were collected from 29 of 72 (40.3%) study sites. In total, 26.1% of the ticks (85.7% of I. ricinus and 14.3% of D. reticulatus) yielded DNA of Borrelia. The difference in the infection rate between I. ricinus and D. reticulatus was significant. Among infected ticks, the most frequent spirochete species were B. lusitaniae (50.0%) and B. afzelii (26.2%), followed by B. spielmanii (9.5%), B. valaisiana (7.1%), B. burgdorferi sensu stricto, (4.8%) and B. miyamotoi (2.4%). No co-infections were found. We did not observe a correlation in the occurrence of Borrelia spirochetes in ticks found in individual study sites that differed in terms of habitat type and height of vegetation. Our findings demonstrate that the Borrelia transmission cycles are active within urban habitats, pointing the need for monitoring of tick-borne pathogens in public green areas. They could serve as guidelines for authorities for the proper management of urban green spaces in a way that may limit tick populations and the potential health risks posed by tick-borne pathogens. [ABSTRACT FROM AUTHOR]

Borrelia, spirochetes transmitted by ticks, are the etiological agents of numerous multisystemic diseases, such as Lyme borreliosis (LB) and tick-borne relapsing fever (TBRF). This study focuses on two surface proteins from two Borrelia subspecies involved in these diseases: CspZ, expressed by Borrelia burgdorferi sensu stricto (also named BbCRASP-2 for complement regulator-acquiring surface protein 2), and the factor H binding A (FhbA), expressed by Borrelia hermsii. Numerous subspecies of Borrelia, including these latter, are able to evade the immune defenses of a variety of potential vertebrate hosts in a number of ways. In this context, previous data suggested that both surface proteins play a role in the immune evasion of both Borrelia subspecies by interacting with key regulators of the alternative pathway of the human complement system, factor H (FH) and FH-like protein 1 (FHL-1). The recombinant proteins, CspZ and FhbA, were expressed in Escherichia coli and purified by one-step metal-affinity chromatography, with yields of 15 and 20 mg or pure protein for 1 L of cultured bacteria, respectively. The purity was evaluated by SDS-PAGE and HPLC and is close to about 95%. The mass of CspZ and FhbA was checked by mass spectrometry (MS). Proper folding of CspZ and FhbA was confirmed by circular dichroism (CD), and their biological activity, namely their interaction with purified FH from human serum (recombinant FH15-20 and recombinant FHL-1), was characterized by SPR. Such a study provides the basis for the biochemical characterization of the studied proteins and their biomolecular interactions which is a necessary prerequisite for the development of new approaches to improve the current diagnosis of LB and TBRF. Key points: • DLS, CD, SEC-MALS, NMR, HPLC, and MS are tools for protein quality assessment • Borrelia spp. possesses immune evasion mechanisms, including human host complement • CspZ and FhbA interact with high affinity (pM to nM) to human FH and rFHL-1 [ABSTRACT FROM AUTHOR]

Relapsing fever (RF), a vector-borne disease caused by Borrelia spp., is characterized by recurring febrile episodes due to repeated bouts of bacteremia. RF spirochetes can be geographically and phylogenetically divided into two distinct groups; Old World RF Borrelia (found in Africa, Asia, and Europe) and New World RF Borrelia (found in the Americas). While RF is a rarely reported disease in the Americas, RF is prevalent in endemic parts of Africa. Despite phylogenetic differences between Old World and New World RF Borrelia and higher incidence of disease associated with Old World RF spirochete infection, genetic manipulation has only been described in New World RF bacteria. Herein, we report the generation of genetic tools for use in the Old World RF spirochete, Borrelia duttonii. We describe methods for transformation and establish shuttle vector- and integration-based approaches for genetic complementation, creating green fluorescent protein (gfp)-expressing B. duttonii strains as a proof of principle. Allelic exchange mutagenesis was also used to inactivate a homolog of the Borrelia burgdorferi p66 gene, which encodes an important virulence factor, in B. duttonii and demonstrate that this mutant was attenuated in a murine model of RF. Finally, the B. duttonii p66 mutant was complemented using shuttle vector- and cis integration-based approaches. As expected, complemented p66 mutant strains were fully infectious, confirming that P66 is required for optimal mammalian infection. The genetic tools and techniques reported herein represent an important advancement in the study of RF Borrelia that allows for future characterization of virulence determinants and colonization factors important for the enzootic cycle of Old World RF spirochetes. Author summary: Relapsing fever is a globally distributed, vector-borne bacterial infection that is characterized by recurring febrile episodes. The causative Borrelia spp. were identified over 100 years ago, but little is known regarding factors required for mammalian infection or vector colonization/transmission. Relapsing fever Borrelia can be geographically and phylogenetically divided into distinct clades; New World, found in the Americas, and Old World, found in Africa, Europe, and Asia. Although Old World relapsing fever Borrelia represent major causes of disease in endemic regions of the world, genetic studies aimed at identifying bacterial gene products required during the tick-vertebrate infectious cycle have focused on the less commonly reported New World relapsing fever Borrelia. Herein, we begin to address this knowledge gap by developing techniques and molecular tools for genetic manipulation of the Old World tick-borne relapsing fever spirochete, Borrelia duttonii. This genetic system will lay the foundation for future studies aimed at identifying bacterial factors required during the enzootic cycle of B. duttonii. [ABSTRACT FROM AUTHOR]

Aim: To describe the aetiology and clinical characteristics of acute peripheral facial palsy (PFP) in children and investigate the utility of the European Federation of Neurological Societies (EFNS) criteria for diagnosing Borrelia‐related PFP (BPFP) based on cerebrospinal fluid (CSF) testing and the Centers for Disease Control and Prevention (CDC) criteria based on serology. Methods: We retrospectively identified children aged <18 years diagnosed with acute PFP between 2014 and 2020. We used the EFNS criteria as the gold standard and the CDC criteria for diagnosing BPFP. Results: Out of 257 children with PFP, 93 (36%) fulfilled the EFNS or CDC criteria for BPFP. We found a discrepancy between the EFNS criteria with CSF testing and the CDC without CSF testing in 27 (14%) of the 190 children with available data. Of the 37 children with PFP and ≥2 symptoms of fever, fatigue, nausea/vomiting or meningeal symptoms, 31 (84%) fulfilled the EFNS criteria for BPFP. Conclusion: Borrelia is a common cause of PFF in children, and its prevalence is higher in children with systemic symptoms. Also, CSF testing did not have decisive management implications in most cases. Therefore, clinical evaluation and Borrelia serology could be the initial steps in the diagnosis of PFP in children. [ABSTRACT FROM AUTHOR]

Ticks are ectoparasites of a wide range of animals and are important vectors of numerous pathogens affecting humans, livestock, and pets. This study investigates possible correlations between selected factors, altitude, soil pH, and a factor called 'amount' (number of ticks examined in pooled samples) on the occurrence of I. ricinus ticks positive for selected tick-borne microorganisms. Questing I. ricinus ticks were collected in 2016 and 2017 across various altitudes, at two mountain ranges in central Slovakia. Tick pools were screened for the presence of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato (Bbsl), Babesia/Theileria spp., Rickettsia spp., and tick-borne encephalitis virus (TBEV) using molecular methods. Regression analysis was employed to evaluate relationships between selected factors and the occurrence of vector-borne microorganisms. This study revealed a statistically significant influence of altitude on the occurrence of A. phagocytophilum; increasing altitude of the sampling site was associated with increased probability of pathogen occurrence. For Babesia/Theileria spp., neither altitude nor soil pH significantly affected pathogen occurrence. The occurrence of Bbsl was notably impacted by both altitude and soil pH; higher altitudes were associated with a decreased probability of pathogen presence, whereas higher soil pH increased the likelihood of pathogen occurrence. The presence of Rickettsia in a pooled sample was not affected by altitude and soil pH, but the 'amount' factor was a significant predictor, increasing the probability of pathogen detection. Neither altitude nor soil pH had a significant impact on TBEV occurrence. The regression models showed moderate goodness-of-fit levels to the data, underscoring their utility in examining the role of altitude and soil pH on pathogen occurrence. However, they explained only a small portion of the overall variance in pathogen occurrence, indicating the presence of other significant factors not covered in this study. [ABSTRACT FROM AUTHOR]

Karachay-Cherkessia is the region in the Russian Federation that has been overlooked in terms of research on the human bacterial pathogens transmitted by ticks. In this study, we investigated the infection status of ticks with Rickettsia, Borrelia, Coxiella burmetii, Anaplasma phagocytophilum, and Ehrlichia chaffeensis/Ehrlichia muris associated with natural focal infections in Karachay-Cherkessia. A total amount of 159 ticks from three species across three genera (Ixodes ricinus, Dermacentor marginatus, Hyalomma scupense) removed from humans were collected for analysis. Additionally, we used 53 individual ticks and 40 tick pools from the vegetation of three species (I. ricinus, D. marginatus, and Rhipicephalus bursa). Screening of the studied pathogens was performed by using commercial qPCR kits. Sanger sequencing utilizing partial sequences of gltA and ompA genes was employed to identify the Rickettsia species. Our findings revealed the presence of DNA from five species within the spotted fever group Rickettsia, namely Rickettsia raoultii, R. slovaca, R. helvetica, R. monacensis, and R. aeschlimannii. Moreover, two distinct genotypes were identified within R. aeschlimannii, R. helvetica, and R. monacensis. Additionally, DNA from Borrelia burgdoferi s.l., B. miyamotoi, and A. phagocytophilum was detected in the tested ticks. This study provides valuable insights into the prevalence and the diversity of bacterial pathogens transmitted by ticks in the Karachay-Cherkessia region. [ABSTRACT FROM AUTHOR]

Emerging and re‐emerging pathogens often stem from zoonotic origins, cycling between humans and animals, and are frequently vectored and maintained by hematophagous arthropod vectors. The efficiency by which these disease agents are successfully transmitted between vertebrate hosts is influenced by many factors, including the host on which a vector feeds. The Lyme disease bacterium Borrelia burgdorferi sensu lato has adapted to survive in complex host environments, vectored by Ixodes ticks, and maintained in multiple vertebrate hosts. The versatility of Lyme borreliae in disparate host milieus is a compelling platform to investigate mechanisms dictating pathogen transmission through complex networks of vertebrates and ticks. Squamata, one of the most diverse clade of extant reptiles, is comprised primarily of lizards, many of which are readily fed upon by Ixodes ticks. Yet, lizards are one of the least studied taxa at risk of contributing to the transmission and life cycle maintenance of Lyme borreliae. In this review, we summarize the current evidence, spanning from field surveillance to laboratory infection studies, supporting their contributions to Lyme borreliae circulation. We also summarize the current understanding of divergent lizard immune responses that may explain the underlying molecular mechanisms to confer Lyme spirochete survival in vertebrate hosts. This review offers a critical perspective on potential enzootic cycles existing between lizard‐tick‐Borrelia interactions and highlights the importance of an eco‐immunology lens for zoonotic pathogen transmission studies. [ABSTRACT FROM AUTHOR]