Lyme borreliosis (LB) is the most prevalent tick-borne human infection in Europe, with increasing incidence during the latest decades. Abundant populations of Ixodes ricinus, the main vector of the causative agent, spirochetes from the Borrelia burgdorferi sensu lato (Bbsl) complex, have been observed in urban and suburban areas of Europe, in general, and Slovakia, particularly. Understanding the spread of infectious diseases is crucial for implementing effective control measures. Global changes affect contact rates of humans and animals with Borrelia-infected ticks and increase the risk of contracting LB. The aim of this study was to investigate spatial and temporal variation in prevalence of Bbsl and diversity of its species in questing I. ricinus from three sites representing urban/suburban, natural and agricultural habitat types in Slovakia. Ixodes ricinus nymphs and adults were collected by dragging the vegetation in green areas of Bratislava town (urban/suburban habitat), in the Small Carpathians Mountains (natural habitat) (south-western Slovakia) and in an agricultural habitat at Rozhanovce in eastern Slovakia. Borrelia presence in ticks was detected by PCR and Bbsl species were identified by restriction fragment length polymorphism (RFLP). Borrelia burgdorferi s.l. species in coinfected ticks were identified by reverse line blot. Significant spatial and temporal variability in prevalence of infected ticks was revealed in the explored habitats. The lowest total prevalence was detected in the urban/suburban habitat, whereas higher prevalence was found in the natural and agricultural habitat. Six Bbsl species were detected by RFLP in each habitat type -B. burgdorferi sensu stricto (s.s.), B. afzelii, B. garinii, B. valaisiana, B. lusitaniae and B. spielmanii. Coinfections accounted for 3% of the total infections, whereby B. kurtenbachii was identified by RLB and sequencing in mixed infection with B. burgdorferi s.s, B. garinii and B. valaisiana. This finding represents the first record of B. kurtenbachii in questing I. ricinus in Slovakia and Europe. Variations in the proportion of Bbsl species were found between nymphs and adults, between years and between habitat types. Spatial variations in prevalence patterns and proportion of Bbsl species were also confirmed between locations within a relatively short distance in the urban habitat. Habitat-related and spatial variations in Borrelia prevalence and distribution of Bbsl species are probably associated with the local environmental conditions and vertebrate host spectrum. Due to the presence of Borrelia species pathogenic to humans, all explored sites can be ranked as areas with high epidemiological risk.

• Keywords
• Borrelia burgdorferi, Ixodes ricinus, Slovakia, prevalence, species,
• Publication type
• Journal Article MeSH

Lyme borreliosis (LB), caused by spirochetes of the Borrelia burgdorferi sensu lato (s.l.) complex, is one of the most common vector-borne zoonotic diseases in Europe. Knowledge about the enzootic circulation of Borrelia pathogens between ticks and their vertebrate hosts is epidemiologically important and enables assessment of the health risk for the human population. In our project, we focused on the following vertebrate species: European hedgehog (Erinaceus europaeus), Northern white-breasted hedgehog (E. roumanicus), Eurasian red squirrel (Sciurus vulgaris), and Common blackbird (Turdus merula). The cadavers of accidentally killed animals used in this study constitute an available source of biological material, and we have confirmed its potential for wide monitoring of B. burgdorferi s.l. presence and genospecies diversity in the urban environment. High infection rates (90% for E. erinaceus, 73% for E. roumanicus, 91% for S. vulgaris, and 68% for T. merula) were observed in all four target host species; mixed infections by several genospecies were detected on the level of individuals, as well as in particular tissue samples. These findings show the usefulness of multiple tissue sampling as tool for revealing the occurrence of several genospecies within one animal and the risk of missing particular B. burgdorferi s.l. genospecies when looking in one organ alone.

• Keywords
• Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Common blackbird, Eurasian red squirrel, European hedgehog, Northern white-breasted hedgehog,
• Publication type
• Journal Article MeSH

• MeSH
• Genes, Bacterial MeSH
• Borrelia burgdorferi genetics   isolation & purification   MeSH
• Host-Pathogen Interactions MeSH
• Ixodidae microbiology   MeSH
• Humans MeSH
• Lyme Disease epidemiology   microbiology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Letter MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Southeastern United States epidemiology   MeSH

BACKGROUND: Out of 20 spirochete species from Borrelia burgdorferi sensu lato (s.l.) complex recognized to date some are considered to have a limited distribution, while others are worldwide dispersed. Among those are Borrelia burgdorferi sensu stricto (s.s.) and Borrelia bissettii which are distributed both in North America and in Europe. While B. burgdorferi s.s. is recognized as a cause of Lyme borreliosis worldwide, involvement of B. bissettii in human Lyme disease was not so definite yet. FINDINGS: Multilocus sequence typing of spirochete isolates originating from residents of Georgia and Florida, USA, revealed the presence of two Borrelia burgdorferi sensu stricto strains highly similar to those from endemic Lyme borreliosis regions of the northeastern United States, and an unusual strain that differed from any previously described in Europe or North America. Based on phylogenetic analysis of eight chromosomally located housekeeping genes divergent strain clustered between Borrelia bissettii and Borrelia carolinensis, two species from the B.burgdorferi s.l. complex, widely distributed among the multiple hosts and vector ticks in the southeastern United States. The genetic distance analysis showed a close relationship of the diverged strain to B. bissettii. CONCLUSIONS: Here, we present the analysis of the first North American human originated live spirochete strain that revealed close relatedness to B. bissettii. The potential of B. bissettii to cause human disease, even if it is infrequent, is of importance for clinicians due to the extensive range of its geographic distribution.

• MeSH
• Borrelia burgdorferi Group MeSH
• Borrelia burgdorferi MeSH
• Borrelia classification   genetics   isolation & purification   MeSH
• Genes, Essential MeSH
• Genotype MeSH
• Humans MeSH
• Lyme Disease diagnosis   microbiology   MeSH
• Molecular Sequence Data MeSH
• Multilocus Sequence Typing * MeSH
• Sequence Analysis, DNA MeSH
• Cluster Analysis MeSH
• Spirochaetales MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Florida MeSH
• Georgia MeSH

BACKGROUND: Analysis of Borrelia burgdorferi ospC types from the southeastern U.S.A. supported the common belief that various ospC types are geographically restricted and host specific. Being widely distributed in the region, the southeastern population of B. burgdorferi is represented by a surprisingly small number of ospC types. Types B, G and H are dominant or common and are invasive, while scarce type L, restricted mostly to the southeastern U.S.A., is believed to rarely if ever cause human Lyme disease. OspC type B and L strains are represented in the region at the same rate, however their distribution among tick vectors and vertebrate hosts is unequal. FINDINGS: Direct diagnostics was used to analyze the ability of B. burgdorferi ospC type L strains to disseminate into host tissues. Mice were infected by subcutaneous injections of B. burgdorferi strains of various ospC types with different invasive capability. Spirochete levels were examined in ear, heart, bladder and joint tissues. Noninfected I. ricinus larvae were fed on infected mice until repletion. Infection rates were determined in molted nymphs. Infected nymphs were then fed on naïve mice, and spirochete transmission from infected nymphs to mice was confirmed. CONCLUSIONS: B. burgdorferi ospC type L strains from the southeastern U.S.A. have comparable potential to disseminate into host tissues as ospC types strains commonly associated with human Lyme disease in endemic European and North American regions. We found no difference in the invasive ability of ospC type B and L strains originated either from tick vectors or vertebrate hosts.

• MeSH
• Antigens, Bacterial genetics   metabolism   MeSH
• Borrelia burgdorferi classification   physiology   MeSH
• Humans MeSH
• Lyme Disease epidemiology   microbiology   MeSH
• Mice MeSH
• Bacterial Outer Membrane Proteins genetics   metabolism   MeSH
• Gene Expression Regulation, Bacterial physiology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Europe epidemiology   MeSH
• Southeastern United States epidemiology   MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• OspC protein MeSH Browser
• Bacterial Outer Membrane Proteins MeSH

BACKGROUND: The controversy surrounding the potential impact of birds in spirochete transmission dynamics and their capacity to serve as a reservoir has existed for a long time. The majority of analyzed bird species are able to infect larval ticks with Borrelia. Dispersal of infected ticks due to bird migration is a key to the establishment of new foci of Lyme borreliosis. The dynamics of infection in birds supports the mixing of different species, the horizontal exchange of genetic information, and appearance of recombinant genotypes. METHODS: Four Borrelia burgdorferi sensu lato strains were cultured from Ixodes minor larvae and four strains were isolated from Ixodes minor nymphs collected from a single Carolina Wren (Thryothorus ludovicianus). A multilocus sequence analysis that included 16S rRNA, a 5S-23S intergenic spacer region, a 16S-23S internal transcribed spacer, flagellin, p66, and ospC separated 8 strains into 3 distinct groups. Additional multilocus sequence typing of 8 housekeeping genes, clpA, clpX, nifS, pepX, pyrG, recG, rplB, and uvrA was used to resolve the taxonomic status of bird-associated strains. RESULTS: Results of analysis of 14 genes confirmed that the level of divergence among strains is significantly higher than what would be expected for strains within a single species. The presence of cross-species recombination was revealed: Borrelia burgdorferi sensu stricto housekeeping gene nifS was incorporated into homologous locus of strain, previously assigned to B. americana. CONCLUSIONS: Genetically diverse Borrelia strains are often found within the same tick or same vertebrate host, presenting a wide opportunity for genetic exchange. We report the cross-species recombination that led to incorporation of a housekeeping gene from the B. burgdorferi sensu stricto strain into a homologous locus of another bird-associated strain. Our results support the hypothesis that recombination maintains a majority of sequence polymorphism within Borrelia populations because of the re-assortment of pre-existing sequence variants. Even if our findings of broad genetic diversity among 8 strains cultured from ticks that fed on a single bird could be the exception rather than the rule, they support the theory that the diversity and evolution of LB spirochetes is driven mainly by the host.

• MeSH
• Borrelia burgdorferi classification   genetics   isolation & purification   MeSH
• Genes, Essential MeSH
• Phylogeny MeSH
• Genetic Variation MeSH
• Ticks microbiology   MeSH
• Molecular Sequence Data MeSH
• Birds microbiology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH

Comparative analysis of ospC genes from 127 Borrelia burgdorferi sensu stricto strains collected in European and North American regions where Lyme disease is endemic and where it is not endemic revealed a close relatedness of geographically distinct populations. ospC alleles A, B, and L were detected on both continents in vectors and hosts, including humans. Six ospC alleles, A, B, L, Q, R, and V, were prevalent in Europe; 4 of them were detected in samples of human origin. Ten ospC alleles, A, B, D, E3, F, G, H, H3, I3, and M, were identified in the far-western United States. Four ospC alleles, B, G, H, and L, were abundant in the southeastern United States. Here we present the first expanded analysis of ospC alleles of B. burgdorferi strains from the southeastern United States with respect to their relatedness to strains from other North American and European localities. We demonstrate that ospC genotypes commonly associated with human Lyme disease in European and North American regions where the disease is endemic were detected in B. burgdorferi strains isolated from the non-human-biting tick Ixodes affinis and rodent hosts in the southeastern United States. We discovered that some ospC alleles previously known only from Europe are widely distributed in the southeastern United States, a finding that confirms the hypothesis of transoceanic migration of Borrelia species.

• MeSH
• Alleles * MeSH
• Antigens, Bacterial genetics   MeSH
• Borrelia burgdorferi genetics   isolation & purification   MeSH
• DNA, Bacterial chemistry   genetics   MeSH
• Genetic Variation MeSH
• Genotype MeSH
• Rodentia microbiology   MeSH
• Ixodes microbiology   MeSH
• Humans MeSH
• Molecular Sequence Data MeSH
• Bacterial Outer Membrane Proteins genetics   MeSH
• Sequence Analysis, DNA MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geographicals
• Europe MeSH
• North America MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• DNA, Bacterial MeSH
• OspC protein MeSH Browser
• Bacterial Outer Membrane Proteins MeSH

The rare ospC allele L was detected in 30% of Borrelia burgdorferi sensu stricto strains cultured from a tick species, Ixodes affinis, and two rodent host species, Peromyscus gossypinus and Sigmodon hispidus, collected in a coastal plain area of Georgia and South Carolina, in the southeastern United States.

• MeSH
• Alleles MeSH
• Antigens, Bacterial genetics   MeSH
• Borrelia burgdorferi genetics   isolation & purification   MeSH
• DNA, Bacterial chemistry   genetics   MeSH
• Genotype MeSH
• Ixodes microbiology   MeSH
• Peromyscus microbiology   MeSH
• Molecular Sequence Data MeSH
• Bacterial Outer Membrane Proteins genetics   MeSH
• Sequence Analysis, DNA MeSH
• Sigmodontinae microbiology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Geographicals
• Southeastern United States MeSH
• Names of Substances
• Antigens, Bacterial MeSH
• DNA, Bacterial MeSH
• OspC protein MeSH Browser
• Bacterial Outer Membrane Proteins MeSH

Borrelia burgdorferi sensu lato (s.l.) complex is a diverse group of worldwide distributed bacteria that includes 18 named spirochete species and a still not named group proposed as genomospecies 2. Descriptions of new species and variants continue to be recognized, so the current number of described species is probably not final. Most of known spirochete species are considered to have a limited distribution. Eleven species from the B. burgdorferi s.l. complex were identified in and strictly associated with Eurasia (B. afzelii, B. bavariensis, B. garinii, B. japonica, B. lusitaniae, B. sinica, B. spielmanii, B. tanukii, B. turdi, B. valaisiana, and B. yangtze), while another 5 (B. americana, B. andersonii, B. californiensis, B. carolinensis, and B. kurtenbachii) were previously believed to be restricted to the USA only. B. burgdorferi sensu stricto (s.s.), B. bissettii, and B. carolinensis share the distinction of being present in both the Old and the New World. Out of the 18 genospecies, 3 commonly and 4 occasionally infect humans, causing Lyme borreliosis (LB) - a multisystem disease that is often referred to as the 'great imitator' due to diversity of its clinical manifestations. Among the genospecies that commonly infect people, i.e. B. burgdorferi s.s., B. afzelii, and B. garinii, only B. burgdorferi s.s. causes LB both in the USA and in Europe, with a wide spectrum of clinical conditions ranging from minor cutaneous erythema migrans (EM) to severe arthritis or neurological manifestations. The epidemiological data from many European countries and the USA show a dramatic increase of the diagnosed cases of LB due to the development of new progressive diagnostic methods during the last decades (Hubálek, 2009). Recently, the definition of the disease has also changed. What was not considered Lyme borreliosis before might be now.

• MeSH
• Arachnid Vectors microbiology   physiology   MeSH
• Borrelia burgdorferi classification   genetics   isolation & purification   pathogenicity   MeSH
• DNA, Bacterial genetics   MeSH
• Species Specificity MeSH
• Phylogeny MeSH
• Phylogeography MeSH
• Genetic Variation MeSH
• Rodentia MeSH
• Ticks microbiology   physiology   MeSH
• Humans MeSH
• Lyme Disease * diagnosis   epidemiology   microbiology   transmission   MeSH
• Polymerase Chain Reaction MeSH
• Birds MeSH
• Bacterial Typing Techniques methods   MeSH
• Public Health MeSH
• Disease Reservoirs MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geographicals
• Asia MeSH
• Europe MeSH
• United States MeSH
• Names of Substances
• DNA, Bacterial MeSH

A group of 16 isolates with genotypic characteristics different from those of known species of the Borrelia burgdorferi sensu lato complex were cultured from ear biopsies of the rodents Peromyscus gossypinus and Neotoma floridana trapped at five localities in South Carolina, USA, and from the tick Ixodes minor feeding on N. floridana. Multilocus sequence analysis of members of the novel species, involving the 16S rRNA gene, the 5S-23S (rrf-rrl) intergenic spacer region and the flagellin, ospA and p66 genes, was conducted and published previously and was used to clarify the taxonomic status of the novel group of B. burgdorferi sensu lato isolates. Phylogenetic analysis based on concatenated sequences of the five analysed genomic loci showed that the 16 isolates clustered together but separately from other species in the B. burgdorferi sensu lato complex. The analysed group therefore represents a novel species, formally described here as Borrelia carolinensis sp. nov., with the type strain SCW-22(T) (=ATCC BAA-1773(T) =DSM 22119(T)).

• MeSH
• Borrelia burgdorferi Group classification   genetics   isolation & purification   MeSH
• DNA, Bacterial genetics   MeSH
• Flagellin genetics   MeSH
• Phylogeny * MeSH
• Ixodes microbiology   MeSH
• DNA, Ribosomal Spacer genetics   MeSH
• Molecular Sequence Data MeSH
• Multilocus Sequence Typing MeSH
• Polymorphism, Restriction Fragment Length MeSH
• RNA, Ribosomal, 16S genetics   MeSH
• Sequence Analysis, DNA MeSH
• Sigmodontinae microbiology   MeSH
• Bacterial Typing Techniques MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geographicals
• South Carolina MeSH
• Names of Substances
• DNA, Bacterial MeSH
• Flagellin MeSH
• DNA, Ribosomal Spacer MeSH
• RNA, Ribosomal, 16S MeSH