In 2005 and 2006, Ixodes ricinus ticks were collected on two slag (waste rock) heaps from coal mines in the Ostrava area (North Moravia/Silesia, Czech Republic), Oskar (site A) and Emma (site B), partially covered by vegetation including trees, and at a control forest site near Hlucín (site C). The mean numbers of L. ricinus nymphs and imagoes flagged per person-hour were high: 35.3 nymphs and 12.7 imagoes, at site A, 23.3 and 26.0, respectively, at site B, and 25.4 and 16.8, respectively, at control site C. Using dark-field microscopy, 100 nymphs and 100 imagoes (50 females and 50 males) from each site were examined for borreliae. The mean prevalence rates of Borrelia burgdorferi sensu lato in nymphs and imagoes were 10.0% and 12.0%, respectively, at site A, 10.0% and 24.0%, respectively, at site B, and 13.0% and 17.0%, respectively, at site C. Differences in the prevalence of borreliae in nymphal and adult ticks from the slag heaps and control site were insignificant, but adult ticks from site B compared to site A contained borreliae significantly more frequently. The mean numbers of nymphs and imagoes infected with borreliae flagged per person-hour were 3.3 and 1.2, respectively at site A, 1.5 and 2.9, respectively, at site B, and 3.1 and 2.6, respectively, at site C. Isolation experiments for borreliae were carried out only in 16 ticks containing higher numbers of borreliae, with eight of these being culture-positive. The cultured borreliae were identified by PCR-RFLP as B. garinii (3 isolates: two from site B, one from site C), B. afzelii (4 isolates: one from site A, three from site B) and B. burgdorferi s.s. (one isolate from site A). Surprisingly, the results suggest that slag heaps, when covered by woody vegetation and frequented by humans, could theoretically pose roughly the same LB transmission risk to humans as common forest biotopes.

• MeSH
• Borrelia burgdorferi komplex izolace a purifikace   MeSH
• ekosystém MeSH
• klíšťata virologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• anglický abstrakt MeSH
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

The Borrelia consists of three groups of species, those of the Lyme borreliosis (LB) group, also known as B. burgdorferi sensu lato (s.l.) and recently reclassified into Borreliella, the relapsing fever (RF) group Borrelia, and a third reptile-associated group of spirochetes. Culture-based methods remain the gold standard for the laboratory detection of bacterial infections for both research and clinical work, as the culture of pathogens from bodily fluids or tissues directly detects replicating pathogens and provides source material for research. Borrelia and Borreliella spirochetes are fastidious and slow growing, and thus are not commonly cultured for clinical purposes; however, culture is necessary for research. This protocol demonstrates the methodology and recipes required to successfully culture LB and RF spirochetes, including all recognized species from B. burgdorferi s.l. complex including B. afzelii, B. americana, B. andersonii, B. bavariensis, B. bissettii/bissettiae, B. burgdorferi sensu stricto (s.s.), B. californiensis, B. carolinensis, B. chilensis, B. finlandensis, B. garinii, B. japonica, B. kurtenbachii, B. lanei, B. lusitaniae, B. maritima, B. mayonii, B. spielmanii, B. tanukii, B. turdi, B. sinica, B. valaisiana, B. yangtzensis, and RFspirochetes, B. anserina, B. coriaceae, B. crocidurae, B. duttonii, B. hermsii, B. hispanica, B. persica, B. recurrentis, and B. miyamotoi. The basic medium for growing LB and RF spirochetes is the Barbour-Stoenner-Kelly (BSK-II or BSK-H) medium, which reliably supports the growth of spirochetes in established cultures. To be able to grow newly isolated Borrelia isolates from tick- or host-derived samples where the initial spirochete number is low in the inoculum, modified Kelly-Pettenkofer (MKP) medium is preferred. This medium also supports the growth of B. miyamotoi. The success of the cultivation of RF spirochetes also depends critically on the quality of ingredients.

• MeSH
• Borrelia burgdorferi komplex * MeSH
• Borrelia burgdorferi * MeSH
• Borrelia * MeSH
• lidé MeSH
• lymeská nemoc * diagnóza   MeSH
• návratná horečka * diagnóza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• audiovizuální média MeSH
• časopisecké články MeSH

The aim of the present study is to compare the prevalence of Borrelia burgdorferi sensu lato (s.l.) and Anaplasma phagocytophilum in questing ticks by using molecular methods from spring to autumn 2007. A total of 526 Ixodes ricinus ticks were collected from vegetation in three different sampling sites, representing an urban area (city park), a suburban area (village) and a natural montane habitat. The prevalence of B. burgdorferi s.l. was 17.3% (7.5% in males, 18.7% in females and 20.2% in nymphs), while 4.4% of ticks (13.1% of males, 3.7% of females and 1.6% of nymphs) tested positive for A. phagocytophilum. We found higher rates of Anaplasma infection in ticks from the urban area (8.6%) than from the suburban (0.8%) and natural (1.6%) habitats in the spring months. The prevalence of Borrelia infection in the urban park increased significantly from spring (14% in March) to autumn (50% in October). The Anaplasma positivity in the urban area in the autumn months (2.2%) was significantly lower than in the spring and summer months (9.6%). The prevalence of A. phagocytophilum was significantly higher in male ticks than in females and nymphs. For B. burgdorferi s.l., the inverse was true. We conclude that infection risks associated with the presence of Anaplasma and Borrelia in ticks in cities may be comparable to those in natural ecosystems or may be even higher. Our results indicate the need for the surveillance of tick-borne pathogens in urban areas.

• Klíčová slova
• Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Ixodes ricinus, PCR, Tick,
• MeSH
• Anaplasma phagocytophilum izolace a purifikace   MeSH
• Borrelia burgdorferi komplex izolace a purifikace   MeSH
• ekosystém MeSH
• Ixodidae růst a vývoj   mikrobiologie   MeSH
• nymfa růst a vývoj   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

BACKGROUND: During the last decades, population densities of Ixodes ricinus and prevalences of Borrelia burgdorferi s.l. have increased in different regions in Europe. In the present study, we determined tick abundance and the prevalence of different Borrelia genospecies in ticks from three sites in the Siebengebirge, Germany, which were already examined in the years 1987, 1989, 2001 and 2003. Data from all investigations were compared. METHODS: In 2007 and 2008, host-seeking I. ricinus were collected by monthly blanket dragging at three distinct vegetation sites in the Siebengebirge, a nature reserve and a well visited local recreation area near Bonn, Germany. In both years, 702 ticks were tested for B. burgdorferi s.l. DNA by nested PCR, and 249 tick samples positive for Borrelia were further genotyped by reverse line blotting. RESULTS: A total of 1046 and 1591 I. ricinus were collected in 2007 and 2008, respectively. In comparison to previous studies at these sites, the densities at all sites increased from 1987/89 and/or from 2003 until 2008. Tick densities and Borrelia prevalences in 2007 and 2008, respectively, were not correlated for all sites and both years. Overall, Borrelia prevalence of all ticks decreased significantly from 2007 (19.5%) to 2008 (16.5%), thus reaching the same level as in 2001 two times higher than in 1987/89 (7.6%). Since 2001, single infections with a Borrelia genospecies predominated in all collections, but the number of multiple infections increased, and in 2007, for the first time, triple Borrelia infections occurred. Prevalences of Borrelia genospecies differed considerably between the three sites, but B. garinii or B. afzelii were always the most dominant genospecies. B. lusitaniae was detected for the first time in the Siebengebirge, also in co-infections with B. garinii or B. valaisiana. CONCLUSIONS: Over the last two centuries tick densities have changed in the Siebengebirge at sites that remained unchanged by human activity since they belong to a nature reserve. Abiotic and biotic conditions most likely favored the host-seeking activity of I. ricinus and the increase of multiple Borrelia infections in ticks. These changes have led to a potential higher risk of humans and animals to be infected with Lyme borreliosis.

• MeSH
• Borrelia burgdorferi komplex klasifikace   genetika   izolace a purifikace   MeSH
• časové faktory MeSH
• DNA bakterií genetika   MeSH
• hustota populace MeSH
• klíště růst a vývoj   mikrobiologie   MeSH
• polymerázová řetězová reakce MeSH
• prevalence MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Německo MeSH
• Názvy látek
• DNA bakterií MeSH

The aim of this study was to determine the seroprevalence of antibodies to B. burgdorferi s.l. in wild small mammals in the Czech Republic and compare sensitivity of PCR and cultivaton. Wild small mammals (n = 691) were trapped in years 2010-2014 in three localities of the Czech Republic. Heart rinses (n = 340) and sera (n = 351) were examined by modified indirect ELISA. Seventy animals were randomly selected for comparison of results of cultivation and PCR. Mean annual antiborelian positivity was 12% with statistical difference (p < 0.05) between Bank Vole (Clethrionomys glareolus) and other six animal species, while there was no statistical difference (p > 0.05) between rodentia and insectivora, gender and localities. The cultivation revealed one positive sample (1.4%), negative in both PCR and ELISA. Method PCR revealed seven positive samples (10%); two of them were simultaneously dubious in ELISA. Eleven animals, negative in cultivation and PCR, had antibodies in ELISA. Method of PCR compared to cultivation seems to be more sensitive for detection of Borrelia.

• Klíčová slova
• Lyme borreliosis, Rodents, Seroprevalence, Spirochetes, Ticks, Vector-borne disease,
• MeSH
• Arvicolinae imunologie   mikrobiologie   MeSH
• Borrelia burgdorferi genetika   růst a vývoj   imunologie   MeSH
• divoká zvířata imunologie   mikrobiologie   MeSH
• ELISA MeSH
• hlodavci imunologie   mikrobiologie   MeSH
• imunoglobulin G krev   MeSH
• imunoglobulin M krev   MeSH
• lymeská nemoc epidemiologie   imunologie   veterinární   MeSH
• polymerázová řetězová reakce MeSH
• protilátky bakteriální krev   MeSH
• savci imunologie   mikrobiologie   MeSH
• senzitivita a specificita MeSH
• séroepidemiologické studie MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• imunoglobulin G MeSH
• imunoglobulin M MeSH
• protilátky bakteriální MeSH

Two different genetic loci, flaB and ospC, were employed to assign genospecies and OspC phylogenetic type to 18 strains isolated from ticks collected in Pisárky, a suburban park in the city of Brno, Czech Republic. The RFLP analysis revealed three different genospecies (B. afzelii, B. garinii, and B. valaisiana). Three samples from the collection contained more than one genospecies. In the other 15 strains, nucleotide sequences of flaB and ospC were determined. The following phylogenetic analysis assigned 12 isolates to genospecies B. garinii and three to B. afzelii. These isolates were further subdivided into seven distinct ospC groups. The most related OspC types were G2, G4, and G5 (B. garinii) and A3 and A8 (B. afzelii).

• Klíčová slova
• Borrelia burgdorferi, Flagellin B, Lyme disease, OspC, Tick,
• MeSH
• antigeny bakteriální genetika   MeSH
• Borrelia burgdorferi komplex klasifikace   MeSH
• DNA bakterií genetika   MeSH
• flagelin genetika   MeSH
• fylogeneze MeSH
• klíště mikrobiologie   MeSH
• lymeská nemoc MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• proteiny vnější bakteriální membrány genetika   MeSH
• sérotypizace * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• antigeny bakteriální MeSH
• DNA bakterií MeSH
• flaB1 protein, bacteria MeSH Prohlížeč
• flagelin MeSH
• OspC protein MeSH Prohlížeč
• proteiny vnější bakteriální membrány MeSH

• MeSH
• Borrelia burgdorferi * MeSH
• divoká zvířata parazitologie   MeSH
• klíště mikrobiologie   MeSH
• lidé MeSH
• lymeská nemoc epidemiologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• dopisy MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Normanské ostrovy epidemiologie   MeSH

Borrelia burgdorferi sensu lato (s.l.) is the etiological agent of Lyme disease, transmitted by ticks of the genus Ixodes Latreille. Diagnosis of Lyme disease in humans is often difficult and a detailed knowledge of the circulation of B. burgdorferi s.l. in tick hosts is therefore fundamental to support clinical procedures. Here we developed a molecular approach for the detection of B. burgdorferi s.l. in North Italian Ixodes ricinus (Linnaeus). The method is based on the amplification of a fragment of the groEL gene, which encodes a heat-shock protein highly conserved among B. burgdorferi s.l. species. The tool was applied in both qualitative and Real-time PCR approaches testing ticks collected in a North Italian area. The obtained results suggest that this new molecular tool could represent a sensitive and specific method for epidemiological studies aimed at defining the distribution of B. burgdorferi s.l. in I. ricinus and, consequently, the exposure risk for humans.

• Klíčová slova
• Ixodes ricinus, Lyme diseases, Real-time PCR., detection, qualitative PCR,
• MeSH
• bakteriální proteiny analýza   MeSH
• Borrelia burgdorferi komplex genetika   izolace a purifikace   MeSH
• chaperon hsp60 analýza   MeSH
• klíště růst a vývoj   mikrobiologie   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• nymfa růst a vývoj   mikrobiologie   MeSH
• sekvenční analýza proteinů MeSH
• sekvenční seřazení MeSH
• senzitivita a specificita MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Itálie MeSH
• Názvy látek
• bakteriální proteiny MeSH
• chaperon hsp60 MeSH

Questing Ixodes ricinus L. (Acari: Ixodidae) ticks were collected on a forest trail that had been completely cleared of shrubs and ground vegetation in winter 2002 and on a nearby control uncleared forest transect in South Moravia (Czech Republic). Samples were collected each May in 2003, 2004 and 2005. Nymphal ticks were 3.4 times, 1.9 times and 1.2 times less frequent on cleared forest than on uncleared forest trails in the three respective years, whereas adult tick abundance was 27.2 times, 4.0 times and 2.2 times lower, respectively. The ticks were examined for borreliae by dark-field microscopy: prevalence of nymphal ticks infected with Borrelia burgdorferi sensu lato (12.6% to 20.0%) did not differ significantly between the cleared and uncleared trail during the 3 years. In conclusion, the habitat modification appeared to result in a decreased abundance of I. ricinus as well as a reduced frequency of infected ticks (and thus indirectly a lower potential risk of Lyme borreliosis), which lasted, however, for only 2 years. Eight cultures of borreliae isolated from the ticks were all identified as the 'ornithophilic' genomic species Borrelia garinii, possibly indicating a greater role of forest birds than that of forest rodents as the hosts of immature I. ricinus in the tick (and borrelial) colonization of the cleared part of the forest.

• MeSH
• arachnida jako vektory mikrobiologie   fyziologie   MeSH
• Borrelia burgdorferi komplex izolace a purifikace   MeSH
• časové faktory MeSH
• hustota populace MeSH
• klíště mikrobiologie   fyziologie   MeSH
• lymeská nemoc epidemiologie   přenos   MeSH
• prevalence MeSH
• rizikové faktory MeSH
• stromy klasifikace   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

Lyme borreliosis and tick-borne encephalitis (TBE) are the most common tick-borne diseases in Germany. We collected Ixodes ricinus ticks from 16 high-risk and four low-risk sites distributed in Lower Bavaria and Upper Palatinate based on the number of human TBE cases recorded at the Robert Koch Institute from 2001 to 2009. A total of 8805 questing ticks (8203 nymphs, 602 adults) were collected in 2010 and examined in pools for the presence of tick-borne encephalitis virus (TBEV) using real-time RT-PCR. Overall TBEV prevalence evaluated as the minimum infection rate (MIR) was 0.26 % (23 positive pools/8805 ticks in 1029 pools). TBEV was detected at seven of the 16 high-risk sites, where MIR ranged from 0.16 to 2.86 %. A total of 3969 ticks were examined by PCR for infection with Borrelia burgdorferi sensu lato (s.l.) targeting the 5 S-23 S rRNA intergenic spacer (IGS) region. IGS nucleotide sequences were used to determine genospecies. Selected positive Borrelia samples were subjected to PCR and sequencing targeting the OspA gene, providing 46 sequences for molecular phylogenetic analysis. Of the 3969 questing ticks, 506 (12.7 %) were positive for B. burgdorferi s.l. Seven B. burgdorferi s.l. genospecies were identified: B. afzelii (41.3 %), B. garinii (19 %), B. valaisiana (13.8 %), B. burgdorferi sensu stricto (11.1 %), B. spielmanii (0.4 %), B. lusitaniae (0.2 %), and Candidatus B. finlandensis (0.6 %). Mixed infections were identified in 13.6 % of the ticks. The rate of infection in questing ticks varied among sites from 5.6 % (72 examined, four positive) to 29.5 % (88 examined, 26 positive). B. burgdorferi s.l. occurred at all 20 sites, whereas TBEV was detected only at the high-risk sites where more human TBE cases were reported compared to low-risk sites.

• Klíčová slova
• Borrelia, Candidatus Borrelia finlandensis, Germany, Ixodes ricinus, Prevalence, Tick-Borne encephalitis,
• MeSH
• Borrelia burgdorferi komplex izolace a purifikace   MeSH
• klíště mikrobiologie   virologie   MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• nymfa MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prevalence MeSH
• viry klíšťové encefalitidy izolace a purifikace   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Německo MeSH