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

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.

• MeSH
• Borrelia * genetika   klasifikace   MeSH
• lidé MeSH
• myši MeSH
• návratná horečka * mikrobiologie   MeSH
• testy genetické komplementace MeSH
• zelené fluorescenční proteiny genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Relapsing fever is caused by the Borrelia species of spirochetes. Louse-borne epidemics of the disease may happen but the endemic disease is generally transmitted to humans by the bite of an infected tick (Ornithodorus). Clinical and laboratory findings of tick-borne relapsing fever in children in the north-west of Iran, Qazvin, were evaluated. This study was conducted from September 1992 to September 2012. Records from 53 cases of tick-borne relapsing fever (TBRF) were reviewed. In positive cases, febrile illness, and spirochetes were recognized in peripheral blood preparations. Of the 53 children younger than 12 years, fifty two percent were male and about one third (34%) of the patients were in the age range of 7-12 years. The disease is recorded through the whole year but its peak occurs during summer (52.8%) and autumn (32.1%). Sixty eight percent of patients were living in urban areas but had frequent travel to rural area. Thirty two percent of the cases were living in rural areas where their dwellings were close to animal shelters. All (100%) of the 53 subjects were febrile. Travellers to the rural areas with high prevalence of the disease should be attentive of the risk of tick-borne relapsing fever and use suitable control measures. Consequently relapsing fever should be considered when patients who live in or have vacationed in north-west of Iran show a recurring febrile illness.

• MeSH
• antibakteriální látky terapeutické užití   MeSH
• ceftriaxon terapeutické užití   MeSH
• cestování statistika a číselné údaje   MeSH
• dítě MeSH
• erythromycin terapeutické užití   MeSH
• kohortové studie MeSH
• kojenec MeSH
• lidé MeSH
• městské obyvatelstvo statistika a číselné údaje   MeSH
• mladiství MeSH
• návratná horečka diagnóza   farmakoterapie   epidemiologie   MeSH
• peniciliny terapeutické užití   MeSH
• předškolní dítě MeSH
• prevalence MeSH
• retrospektivní studie MeSH
• věkové rozložení MeSH
• venkovské obyvatelstvo statistika a číselné údaje   MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Írán MeSH

Introduction: We developed a new simple method to assess the composition of proteinaceous components in the saliva of Ornithodoros moubata, the main vehicle for pathogen transmission and a likely source of bioactive molecules acting at the tick-vertebrate host interface. To collect naturally expectorated saliva from the ticks we employed an artificial membrane feeding technique using a simple, chemically defined diet containing phagostimulants and submitted native saliva samples collected in this way for liquid chromatography-mass spectrometry (LC-MS) analysis. These experiments were conducted with groups of uninfected ticks as well as with O. moubata infected with B. duttonii. The ticks exhibited a fair feeding response to the tested diet with engorgement rates reaching as high as 60-100% of ticks per feeding chamber. The LC-MS analysis identified a total of 17 and 15 proteins in saliva samples from the uninfected and infected O. moubata nymphs, respectively. Importantly, the analysis was sensitive enough to detect up to 9 different proteins in the samples of saliva containing diet upon which as few as 6 nymphal ticks fed during the experiments. Some of the proteins recognized in the analysis are well known for their immunomodulatory activity in a vertebrate host, whereas others are primarily thought of as structural or "housekeeping" proteins and their finding in the naturally expectorated tick saliva confirms that they can be secreted and might serve some functions at the tick-host interface. Most notably, some of the proteins that have long been suspected for their importance in the vector-pathogen interactions of Borrelia spirochetes were detected only in the samples from infected ticks, suggesting that their expression was altered by the persistent colonization of the tick's salivary glands by spirochetes. The simple method described herein is an important addition to the toolbox available to study the vector-host-pathogen interactions in the rapidly feeding soft ticks.

• MeSH
• Argasidae * MeSH
• Borrelia * fyziologie   MeSH
• Ornithodoros * MeSH
• sliny MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Borrelia species fall into two groups, the Borrelia burgdorferi sensu lato (Bbsl) complex, the cause of Lyme borreliosis (also known as Lyme disease), and the relapsing fever group. Both groups exhibit inter- and intraspecies diversity and thus have variations in both clinical presentation and diagnostic approaches. A further layer of complexity is derived from the fact that ticks may carry multiple infectious agents and are able to transmit them to the host during blood feeding, with potential overlapping clinical manifestations. Besides this, pathogens like Borrelia have developed strategies to evade the host immune system, which allows them to persist within the host, including humans. Diagnostics can be applied at different times during the clinical course and utilize sample types, each with their own advantages and limitations. These differing methods should always be considered in conjunction with potential exposure and compatible clinical features. Throughout this review, we aim to explore different approaches providing the reader with an overview of methods appropriate for various situations. This review will cover human pathogenic members of Bbsl and relapsing fever borreliae, including newly recognized Borrelia miyamotoi spirochetes.

• MeSH
• Borrelia klasifikace   izolace a purifikace   MeSH
• členovci - vektory mikrobiologie   MeSH
• klíšťata mikrobiologie   MeSH
• lidé MeSH
• lymeská nemoc diagnóza   MeSH
• návratná horečka diagnóza   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Borrelia miyamotoi patří mezi emergentní patogeny přenášené klíšťaty a fylogeneticky náleží mezi spirochety způsobující tzv. návratné horečky. Je primárně přenášena klíšťaty komplexu Ixodes ricinus podobně jako borrelie způsobující Lymeskou borreliózu. Hostiteli jsou především drobní hlodavci. Je široce rozšířena v mírném pásu severní polokoule s konstantní nízkou prevalencí v klíšťatech v řádu jednotek procent. Dosud byly popsány více než dvě stovky humánních případů onemocnění včetně pěti případů meningoencefalitidy u imunokompromitovaných pacientů. Onemocnění se vyznačuje nespecifickými příznaky, především horečkou, únavou, zimnicí, bolestmi hlavy, svalů a kloubů. Léčba je antibiotická. Diagnostika zahrnuje především PCR a sérologické metody. Přehledová práce přináší současné poznatky o taxonomii, ekologii vektorů a rezervoárových hostitelů, geografickém rozšíření, diagnostice a léčbě onemocnění způsobené spirochetou B. miyamotoi. Práce současně zdůrazňuje potřebu správné etiologie onemocnění a upozorňuje na možnou záměnu s lymeskou borreliózou či lidskou granulocytární anaplazmózou.

Borrelia miyamotoi is an emerging tick-borne pathogen phylogenetically belonging to spirochaetes causing relapsing fever. It is primarily transmitted by ticks from the Ixodes ricinus complex, similarly to borreliae causing Lyme borreliosis. Small rodents can serve as reservoir hosts. It is widespread in mild climate areas of the northern hemisphere, with constant low prevalence in ticks, in the range of units of percent. To date more than 200 human cases have been described including five cases of meningoencephalitis in immunocompromised patients. Clinical features of illness are non-specific, characterized by fever, fatigue, chills, headaches, muscles and joint pains. It can be treated with antibiotics. The diagnostic approach includes mainly PCR and serological methods. This review summarizes current knowledge on B. miyamotoi with an emphasis on taxonomy, ecology of vectors and reservoir hosts, geographical distribution, diagnosis and treatment of the disease. The review also highlights the need for an accurate determination of the etiology of the disease and its differentiation from Lyme borreliosis and human granulocytic anaplasmosis.

• Klíčová slova
• Borrelia miyamotoi,
• MeSH
• Borrelia burgdorferi MeSH
• Borrelia MeSH
• infekce bakteriemi rodu Borrelia * diagnóza   přenos   MeSH
• lidé MeSH
• lymeská nemoc diagnóza   přenos   MeSH
• návratná horečka diagnóza   přenos   MeSH
• nemoci přenášené klíšťaty diagnóza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Borrelia miyamotoi patří mezi emergentní patogeny přenášené klíšťaty komplexu Ixodes ricinus. Jedná se o spirochetu způsobující onemocnění podobné návratné horečce s rozšířením v mírném pásmu severní polokoule. Předložená studie dokumentuje prevalenci B. miyamotoi v klíšťatech Ixodes ricinus ve 4 lokalitách Jihomoravského kraje. Spirocheta B. miyamotoi byla zaznamenána celkem v 10 vzorcích, ve všech 4 lokalitách a ve všech stadiích klíšťat I. ricinus. Výskyt B. miyamotoi v klíšťatech na území České republiky představuje potenciální riziko pro veřejné zdraví.

Borrelia miyamotoi is an emerging tick-borne pathogen. The vector is ticks of the Ixodes ricinus complex, causing relapsing fever- like disease and widespread in the temperate zone of the Northern Hemisphere. The present study documents the prevalence of B. miyamotoi in Ixodes ricinus ticks at four sites of the South Moravian Region. The spirochete B. miyamotoi was detected in a total of 10 samples from all four sites and all instars. The occurrence of B. miyamotoi in ticks in the Czech Republic poses a potential risk to public health.

• MeSH
• Borrelia izolace a purifikace   MeSH
• epidemiologické studie MeSH
• klíště mikrobiologie   MeSH
• lidé MeSH
• návratná horečka * epidemiologie   MeSH
• nemoci přenášené klíšťaty epidemiologie   MeSH
• prevalence MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Česká republika MeSH

PURPOSE: For simultaneous detection of Borrelia miyamotoi (relapsing fever spirochete) and Borrelia burgdorferi sensu lato, we have developed a duplex real-time PCR targeting the flagellin gene (flaB; p41), a locus frequently used in routine diagnostic PCR for B. burgdorferi s.l. detection. METHODS: Primers and probes were designed using multiple alignments of flaB sequences of B. miyamotoi and B. burgdorferi s.l. species. The sensitivity and specificity of primers and probes were determined using serial dilutions (ranging from 10(4) to 10(-1)) of B. miyamotoi and B. burgdorferi s.l. DNA and of several species of relapsing fever spirochetes. Conventional PCR on recG and glpQ and sequencing of p41 PCR products were used to confirm the species assignment. RESULTS: The detection limit of both singleplex and duplex PCR was 10 genome equivalents except for B. spielmanii and two B. garinii genotypes which showed a detection limit of 10(2) genome equivalents. There was no cross reactivity of the B. miyamotoi primers/probes with B. burgdorferi s.l. DNA, while the B. burgdorferi s.l. primer/probe generated a signal with B. hermsii DNA. Out of 2341 Ixodes ricinus ticks from Germany and Slovakia that were screened simultaneously for the presence of B. miyamotoi and B. burgdorferi s.l., 52 were positive for B. miyamotoi and 276 for B. burgdorferi s.l., denoting an average prevalence of 2.2% for B. miyamotoi and 11.8% for B. burgdorferi s.l., and B. miyamotoi DNA was also detectable by PCR using artificial clinical samples. CONCLUSION: The duplex real-time PCR developed here represents a method that permits simultaneous detection and differentiation of B. burgdorferi s.l. and B. miyamotoi in environmental and potentially clinical samples.

• MeSH
• Borrelia klasifikace   genetika   MeSH
• DNA primery genetika   MeSH
• flagelin genetika   MeSH
• klíště mikrobiologie   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• oligonukleotidové sondy genetika   MeSH
• senzitivita a specificita MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Německo MeSH
• Slovenská republika MeSH

• Publikační typ
• atlasy MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• cestovní a tropická medicína

The life cycle of spirochetes of the genus Borrelia includes complex networks of vertebrates and ticks. The tripartite association of Borrelia-vertebrate-tick has proved ecologically successful for these bacteria, which have become some of the most prominent tick-borne pathogens in the northern hemisphere. To keep evolutionary pace with its double-host life history, Borrelia must adapt to the evolutionary pressures exerted by both sets of hosts. In this review, we attempt to reconcile functional, phylogenetic, and ecological perspectives to propose a coherent scenario of Borrelia evolution. Available empirical information supports that the association of Borrelia with ticks is very old. The major split between the tick families Argasidae-Ixodidae (dated some 230-290 Mya) resulted in most relapsing fever (Rf) species being restricted to Argasidae and few associated with Ixodidae. A further key event produced the diversification of the Lyme borreliosis (Lb) species: the radiation of ticks of the genus Ixodes from the primitive stock of Ixodidae (around 217 Mya). The ecological interactions of Borrelia demonstrate that Argasidae-transmitted Rf species remain restricted to small niches of one tick species and few vertebrates. The evolutionary pressures on this group are consequently low, and speciation processes seem to be driven by geographical isolation. In contrast to Rf, Lb species circulate in nested networks of dozens of tick species and hundreds of vertebrate species. This greater variety confers a remarkably variable pool of evolutionary pressures, resulting in large speciation of the Lb group, where different species adapt to circulate through different groups of vertebrates. Available data, based on ospA and multilocus sequence typing (including eight concatenated in-house genes) phylogenetic trees, suggest that ticks could constitute a secondary bottleneck that contributes to Lb specialization. Both sets of adaptive pressures contribute to the resilience of highly adaptable meta-populations of bacteria.

• MeSH
• biologická adaptace MeSH
• biologická evoluce * MeSH
• Borrelia klasifikace   fyziologie   MeSH
• infekce přenášené vektorem * MeSH
• interakce hostitele a patogenu MeSH
• klíšťata mikrobiologie   MeSH
• lidé MeSH
• lymeská nemoc mikrobiologie   přenos   MeSH
• selekce (genetika) MeSH
• zdroje nemoci * mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH