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.

• Klíčová slova
• Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Common blackbird, Eurasian red squirrel, European hedgehog, Northern white-breasted hedgehog,
• Publikační typ
• časopisecké články MeSH

The class Alphaproteobacteria is comprised of a diverse assemblage of Gram-negative bacteria that includes organisms of varying morphologies, physiologies and habitat preferences many of which are of clinical and ecological importance. Alphaproteobacteria classification has proved to be difficult, not least when taxonomic decisions rested heavily on a limited number of phenotypic features and interpretation of poorly resolved 16S rRNA gene trees. Despite progress in recent years regarding the classification of bacteria assigned to the class, there remains a need to further clarify taxonomic relationships. Here, draft genome sequences of a collection of genomes of more than 1000 Alphaproteobacteria and outgroup type strains were used to infer phylogenetic trees from genome-scale data using the principles drawn from phylogenetic systematics. The majority of taxa were found to be monophyletic but several orders, families and genera, including taxa recognized as problematic long ago but also quite recent taxa, as well as a few species were shown to be in need of revision. According proposals are made for the recognition of new orders, families and genera, as well as the transfer of a variety of species to other genera and of a variety of genera to other families. In addition, emended descriptions are given for many species mainly involving information on DNA G+C content and (approximate) genome size, both of which are confirmed as valuable taxonomic markers. Similarly, analysis of the gene content was shown to provide valuable taxonomic insights in the class. Significant incongruities between 16S rRNA gene and whole genome trees were not found in the class. The incongruities that became obvious when comparing the results of the present study with existing classifications appeared to be caused mainly by insufficiently resolved 16S rRNA gene trees or incomplete taxon sampling. Another probable cause of misclassifications in the past is the partially low overall fit of phenotypic characters to the sequence-based tree. Even though a significant degree of phylogenetic conservation was detected in all characters investigated, the overall fit to the tree varied considerably.

• Klíčová slova
• G+C content, Genome BLAST Distance Phylogeny, chemotaxonomy, genome size, morphology, phylogenetic systematics, phylogenomics,
• Publikační typ
• časopisecké články 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 klasifikace   genetika   izolace a purifikace   MeSH
• esenciální geny MeSH
• fylogeneze MeSH
• genetická variace MeSH
• klíšťata mikrobiologie   MeSH
• molekulární sekvence - údaje MeSH
• ptáci mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural 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
• arachnida jako vektory mikrobiologie   fyziologie   MeSH
• Borrelia burgdorferi klasifikace   genetika   izolace a purifikace   patogenita   MeSH
• DNA bakterií genetika   MeSH
• druhová specificita MeSH
• fylogeneze MeSH
• fylogeografie MeSH
• genetická variace MeSH
• hlodavci MeSH
• klíšťata mikrobiologie   fyziologie   MeSH
• lidé MeSH
• lymeská nemoc * diagnóza   epidemiologie   mikrobiologie   přenos   MeSH
• polymerázová řetězová reakce MeSH
• ptáci MeSH
• techniky typizace bakterií metody   MeSH
• veřejné zdravotnictví MeSH
• zdroje nemoci MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geografické názvy
• Asie MeSH
• Evropa MeSH
• Spojené státy americké MeSH
• Názvy látek
• DNA bakterií 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 komplex klasifikace   genetika   izolace a purifikace   MeSH
• DNA bakterií genetika   MeSH
• flagelin genetika   MeSH
• fylogeneze * MeSH
• klíště mikrobiologie   MeSH
• mezerníky ribozomální DNA genetika   MeSH
• molekulární sekvence - údaje MeSH
• multilokusová sekvenční typizace MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• Sigmodontinae mikrobiologie   MeSH
• techniky typizace bakterií MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geografické názvy
• Jižní Karolína MeSH
• Názvy látek
• DNA bakterií MeSH
• flagelin MeSH
• mezerníky ribozomální DNA MeSH
• RNA ribozomální 16S MeSH

Approximately 118 Borrelia isolates were cultured from a variety of rodents, birds, and ticks collected in the southern United States. In addition to a highly diverse group of Borrelia bissettii strains and a homogenous group of Borrelia burgdorferi sensu stricto strains, a group of 16 isolates with unusual characteristics was found. The isolates were cultured from ear biopsy samples of the rodents Peromyscus gossypinus and Neotoma floridana trapped at five localities in South Carolina. A multilocus sequence analysis of the rrf-rrl intergenic spacer, 16S rRNA, fla, ospA, and p66 genes were used to clarify the taxonomic status of the new group of B. burgdorferi sensu lato isolates. Thirteen species of the B. burgdorferi sensu lato complex were used as controls. Unique restriction fragment length polymorphism patterns of the rrf-rrl intergenic spacer region and fla gene were recognized. Unique signature nucleotides were also found in the 16S rRNA gene. A phylogenetic analysis shows that the 16 new isolates cluster together but separately from the other species in the B. burgdorferi sensu lato complex. Our data strongly support the recognition of the 16 isolates as a new B. burgdorferi sensu lato species. We propose to name this genospecies "Borrelia carolinensis" with respect to the place of its currently known geographic location.

• MeSH
• bakteriální proteiny genetika   MeSH
• bakteriální RNA genetika   MeSH
• Borrelia burgdorferi komplex klasifikace   izolace a purifikace   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• geny rRNA MeSH
• křeček rodu Peromyscus mikrobiologie   MeSH
• mezerníky ribozomální DNA chemie   genetika   MeSH
• molekulární sekvence - údaje MeSH
• ribozomální DNA chemie   genetika   MeSH
• RNA ribozomální 16S genetika   MeSH
• sekvenční analýza DNA MeSH
• sekvenční homologie nukleových kyselin MeSH
• shluková analýza MeSH
• Sigmodontinae mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, P.H.S. MeSH
• Geografické názvy
• jihovýchod USA MeSH
• Názvy látek
• bakteriální proteiny MeSH
• bakteriální RNA MeSH
• DNA bakterií MeSH
• mezerníky ribozomální DNA MeSH
• ribozomální DNA MeSH
• RNA ribozomální 16S MeSH

The genotype of Borrelia burgdorferi sensu lato was detected in 371 out of 1244 ticks. Borrelia determination was based on partial sequencing of the 16S rRNA gene and real-time polymerase chain reactions for identification and quantitation of ospA and recA genes. Different Borrelia spp. were identified; B. garinii in 40% ticks followed by B. afzelii (36.3%), B. burgdorferi sensu stricto (12.9%), B. valaisiana (3.5%), B. lusitaniae (0.8%), B. bissettii (0.5%) and B. miyamotoi-like (0.5%). Cultivation of 30 borrelia strains in BSK-H medium, among them B. valaisiana, B. bissettii-like and B. miyamotoi-like strains was unique in Czechia. Calibrated microfluidic-based quantification showed differences in the concentration of the nucleic acids and molar mass of the outer surface proteins of different Borrelia spp. with standard sensitivity and specificity and was helpful for their identification. The outer surface protein OspA was absent in B. miyamotoi-like and the OspB protein in B. valaisiana, B. lusitaniae and in three subtypes of B. garinii.

• MeSH
• antigeny povrchové chemie   genetika   MeSH
• bakteriální vakcíny chemie   genetika   MeSH
• Borrelia burgdorferi komplex genetika   izolace a purifikace   MeSH
• DNA bakterií chemie   genetika   MeSH
• fylogeneze MeSH
• genetická variace MeSH
• klíště mikrobiologie   MeSH
• lipoproteiny chemie   genetika   MeSH
• mikrofluidní analytické techniky MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• proteiny vnější bakteriální membrány chemie   genetika   MeSH
• RecA-rekombinasy chemie   genetika   MeSH
• RNA ribozomální 16S chemie   genetika   MeSH
• sekvence nukleotidů MeSH
• sekvenční seřazení MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny povrchové MeSH
• bakteriální vakcíny MeSH
• DNA bakterií MeSH
• lipoproteiny MeSH
• OspA protein MeSH Prohlížeč
• proteiny vnější bakteriální membrány MeSH
• RecA-rekombinasy MeSH
• RNA ribozomální 16S MeSH

In Europe the Borrelia burgdorferi sensu lato complex is represented by five distinct genospecies: Borrelia burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia valaisiana, and Borrelia lusitaniae. These taxonomic entities are known to differ in their specific associations with vertebrate hosts and to provoke distinct clinical manifestations in human patients. However, exceptions to these rules have often been observed, indicating that strains belonging to a single genospecies may be more heterogeneous than expected. It is, therefore, important to develop alternative identification tools which are able to distinguish Borrelia strains not only at the specific level but also at the intraspecific level. DNA from a sample of 370 Ixodes ricinus ticks collected in the Czech Republic was analyzed by PCR for the presence of a approximately 230-bp fragment of the rrfA-rrlB intergenic spacer of Borrelia spp. A total of 20.5% of the ticks were found to be positive. The infecting genospecies were identified by analyzing the amplified products by the restriction fragment length polymorphism (RFLP) method with restriction enzyme MseI and by single-strand conformation polymorphism (SSCP) analysis. The two methods were compared, and PCR-SSCP analysis appeared to be a valuable tool for rapid identification of spirochetes at the intraspecific level, particularly when large samples are examined. Furthermore, by using PCR-SSCP analysis we identified a previously unknown Borrelia genotype, genotype I-77, which would have gone unnoticed if RFLP analysis alone had been used.

• MeSH
• Borrelia burgdorferi komplex klasifikace   genetika   MeSH
• DNA bakterií genetika   izolace a purifikace   MeSH
• fylogeneze MeSH
• genetická variace * MeSH
• genotyp MeSH
• klíště mikrobiologie   MeSH
• lidé MeSH
• mezerníky ribozomální DNA genetika   MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce metody   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• polymorfismus konformace jednovláknové DNA * MeSH
• sekvenční analýza DNA MeSH
• techniky typizace bakterií MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• DNA bakterií MeSH
• mezerníky ribozomální DNA MeSH

Activities of 19 enzymes were tested by the API ZYM system in 13 strains of Borrelia burgdorferi sensu lato (B. burgdorferi sensu stricto, B. afzelii, B. garinii, B. lusitaniae, B. valaisiana) grown in liquid BSK-H medium supplemented with rabbit serum. All strains produced acid phosphatase, esterase (C4), esterase-lipase (C8), leucine arylamidase and naphthol-AS-BI-phosphohydrolase. Nine strains also produced alkaline phosphatase, and three strains produced alpha-glucosidase. The API ZYM system probably cannot be used for differentiation between B. burgdorferi sensu lato genomospecies.

• MeSH
• alfa-glukosidasy metabolismus   MeSH
• alkalická fosfatasa metabolismus   MeSH
• Borrelia burgdorferi komplex enzymologie   MeSH
• esterasy metabolismus   MeSH
• fosfatasy metabolismus   MeSH
• kyselá fosfatasa metabolismus   MeSH
• leucylaminopeptidasa metabolismus   MeSH
• lipasa metabolismus   MeSH
• lymeská nemoc mikrobiologie   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• alfa-glukosidasy MeSH
• alkalická fosfatasa MeSH
• esterasy MeSH
• fosfatasy MeSH
• kyselá fosfatasa MeSH
• leucylaminopeptidasa MeSH
• lipasa MeSH

A total of 2816 unfed adults nymphs of Ixodes ricinus ticks were collected from vegetation in Kosice (Eastern Slovakia) from 1994 to 1997. Prevalence of Borrelia burgdorferi s. l. in I. ricinus ticks, detected by dark field microscopy, varied and depended upon the year and the habitat of the collected ticks. The lowest prevalence was observed in 1994 (4.8%). During 1995 it increased to 17.2% and during the next two years decreased to 15.5% and 14.2%. The rate of infection varied from 2.1 to 23.3% within 10 examined habitats of the Kosice area. A different value of relative density of ticks was observed in various habitats. It ranged from 9-212 ticks per collecting hour within one flagged area (600 m2) which is 1.5-35.5 ticks per 100 m2. Eight isolates were obtained from the infected ticks. Electrophoresis and immunoblotting with 6 monoclonal antibodies were used for the identification of Borrelia strains. Three tick isolates were identified as B. burgdorferi s. s. and the other three isolates were found to be B. garinii. One strain reacted as a mixed culture of B. burgdorferi s. s., and B. garinii. The strain originated from the Vihorlat Mountains habitat and was detected by PCR-SSCP as B. burgdorferi s. s. with a small amount of B. afzelii. The obtained results emphasize the epidemiological importance not only of B. garinii and B. afzelii but also of B. burgdorferi s. s. in Central Europe.

• MeSH
• Borrelia burgdorferi komplex izolace a purifikace   MeSH
• Borrelia burgdorferi * MeSH
• Borrelia klasifikace   izolace a purifikace   MeSH
• elektroforéza MeSH
• klíště parazitologie   MeSH
• polymerázová řetězová reakce MeSH
• prevalence MeSH
• western blotting MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Slovenská republika MeSH