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Sample records for borrelia burgdorferi serve

  1. Borrelia burgdorferi Infections in the United States

    OpenAIRE

    Heymann, Warren R.; Ellis, Dana L.

    2012-01-01

    It is becoming increasingly evident that the clinical presentation of infection with Borrelia burgdorferi varies greatly between different parts of the world. A growing number of European and Asian isolates of Lyme borreliae, differing from the American strain of Borrelia burgdorferi, have been identified in several different disorders. In light of the increasing number of reports describing an association between various cutaneous disorders and infection with Borrelia burgdorferi and the con...

  2. Erythromycin Resistance in Borrelia burgdorferi

    OpenAIRE

    Terekhova, Darya; Sartakova, Marina L.; Wormser, Gary P.; Schwartz, Ira; Cabello, Felipe C.

    2002-01-01

    Susceptibility testing of laboratory strains and clinical isolates of Borrelia burgdorferi indicates that resistance to erythromycin is present in them. Evaluation of the MICs, minimal bactericidal concentrations, and kinetics of bacterial killing of erythromycin suggests that this resistance is increased by preexposure to the antibiotic, is dependent on inoculum size, and may be the result of selection of subpopulations of bacterial cells with increased resistance.

  3. Polyneuritis cranialis associated with Borrelia burgdorferi.

    OpenAIRE

    Schmutzhard, E; Stanek, G.; Pohl, P.

    1985-01-01

    Three patients with classical idiopathic polyneuritis cranialis, in whom no other aetiology could be detected, were examined serologically, by means of indirect immunofluorescence test, for antibodies (IgM and IgG) against Borrelia burgdorferi, the aetiologic agent of Lyme disease. In each case polyneuritis cranialis was caused by infection with Borrelia burgdorferi. Therapy with penicillin proved to be effective.

  4. Enzyme activities of Borrelia burgdorferi sensu lato

    Czech Academy of Sciences Publication Activity Database

    Heroldová, M.; Němec, M.; Hubálek, Zdeněk; Halouzka, Jiří

    2001-01-01

    Roč. 46, č. 3 (2001), s. 179-182. ISSN 0015-5632 R&D Projects: GA ČR GA206/00/1204 Institutional research plan: CEZ:AV0Z6093917 Keywords : Borrelia burgdorferi * Lyme disease Subject RIV: EE - Microbiology, Virology Impact factor: 0.776, year: 2001

  5. Borrelia burgdorferi tissue morphologies and imaging methodologies.

    Science.gov (United States)

    MacDonald, A B

    2013-08-01

    This manuscript offers an image presentation of diverse forms of Borrelia burgdorferi spirochetes which are not spiral or corkscrew shaped. Explanations are offered to justify the legitimacy of tissue forms of Borrelia which may confuse the inexperienced microscopic examiner and which may lead to the misdiagnosis of non-spiral forms as artifacts. Images from the author's personal collection of Borrelia burgdorferi images and a few select images of Borrelia burgdorferi from the peer-reviewed published literature are presented. A commentary justifying each of the image profiles and a survey of the imaging modalities utilized provides the reader with a frame of reference. Regularly spiraled Borrelia are rarely seen in solid tissues. A variety of straightened, undulating, and clipped-off profiles are demonstrated, and the structural basis for each image is explained. Tissue examination is a diagnostic tool and a quality control for judging the eradication or the persistence of borreliosis following attempts to eradicate the infection with antibiotic therapy. The presence or absence of chronic Lyme borreliosis may be objectively adjudicated by tissue examinations which demonstrate or which fail to show pathogenic microbes in patients who have received a full course of antibiotics. PMID:23479042

  6. Recombinant constructs of Borrelia burgdorferi

    Energy Technology Data Exchange (ETDEWEB)

    Dattwyler, Raymond J. (Setauket, NY); Gomes-Solecki, Maria J. C. (New York, NY); Luft, Benjamin J. (East Setauket, NY); Dunn, John J.(Bellport, NY)

    2007-02-20

    Novel chimeric nucleic acids, encoding chimeric Borrelia proteins comprising OspC or an antigenic fragment thereof and OspA or an antigenic fragment thereof, are disclosed. Chimeric proteins encoded by the nucleic acid sequences are also disclosed. The chimeric proteins are useful as vaccine immunogens against Lyme borreliosis, as well as for immunodiagnostic reagents.

  7. DNA Microarray Assessment of Putative Borrelia burgdorferi Lipoprotein Genes

    OpenAIRE

    Liang, Fang Ting; Nelson, F. Kenneth; Fikrig, Erol

    2002-01-01

    A DNA microarray containing fragments of 137 Borrelia burgdorferi B31 putative lipoprotein genes was used to examine Lyme disease spirochetes. DNA from B. burgdorferi sensu stricto B31, 297, and N40; Borrelia garinii IP90; and Borrelia afzelii P/Gau was fluorescently labeled and hybridized to the microarray, demonstrating the degree to which the individual putative lipoprotein genes were conserved among the genospecies. These data show that a DNA microarray can globally examine the genes enco...

  8. Identification of superoxide dismutase activity in Borrelia burgdorferi.

    OpenAIRE

    Whitehouse, C. A.; Williams, L. R.; Austin, F E

    1997-01-01

    Infective and noninfective strains of Borrelia burgdorferi, along with Borrelia afzelii and Borrelia garinii, possessed a single iron-containing superoxide dismutase (SOD). None of the Lyme disease spirochetes tested possessed catalase or peroxidase activities. The borrelial SOD was not inducible by growth with increased oxygen concentrations and thus appeared to be produced constitutively.

  9. Infections with Ehrlichia canis and Borrelia burgdorferi in a dog

    Directory of Open Access Journals (Sweden)

    Tsachev Ilia

    2007-01-01

    Full Text Available A clinical case of Ehrlichia canis and Borrelia burgdorferi infections in a 5year-old male German Shepherd is described. Clinical, serological, necropsy and histopathological examinations supporting the diagnosis have been performed.

  10. Infections with Ehrlichia canis and Borrelia burgdorferi in a dog

    OpenAIRE

    Tsachev Ilia; Simeonov R.; Petrov Vladimir

    2007-01-01

    A clinical case of Ehrlichia canis and Borrelia burgdorferi infections in a 5year-old male German Shepherd is described. Clinical, serological, necropsy and histopathological examinations supporting the diagnosis have been performed.

  11. Evaluation of a New Culture Medium for Borrelia burgdorferi

    OpenAIRE

    Marques, Adriana R.; Stock, Frida; Gill, Vee

    2000-01-01

    We evaluated the new MPM medium for the growth of Borrelia burgdorferi. All 18 blood samples from 17 patients with Lyme disease were negative. Growth studies showed that by day 4, most organisms in MPM were not viable. Our results reinforce the use of BSK medium as the primary choice for growing B. burgdorferi.

  12. Decorin Binding by DbpA and B of Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi Sensu Stricto

    OpenAIRE

    Salo, Jemiina; Loimaranta, Vuokko; Lahdenne, Pekka; Viljanen, Matti K.; Hytönen, Jukka

    2011-01-01

    Background. Decorin adherence is crucial in the pathogenesis of Lyme borreliosis. Decorin-binding proteins (Dbp) A and B are the adhesins that mediate this interaction. DbpA and B of Borrelia garinii, Borrelia afzelii, and Borrelia burgdorferi sensu stricto (ss) differ in their amino acid sequence, but little attention has been paid to the potential difference in their decorin binding.

  13. Lipid Exchange between Borrelia burgdorferi and Host Cells

    OpenAIRE

    Crowley, Jameson T; Toledo, Alvaro M; LaRocca, Timothy J.; Coleman, James L.; Erwin London; Benach, Jorge L.

    2013-01-01

    Author Summary Lyme disease, the most prevalent arthropod-borne disease in North America, is caused by the spirochete Borrelia burgdorferi. Cholesterol is a significant component of the B. burgdorferi membrane lipids, and is processed to make cholesterol-glycolipids. Our interest in the presence of cholesterol in B. burgdorferi recently led to the identification and characterization of eukaryotic-like lipid rafts in the spirochete. The presence of free cholesterol and cholesterol-glycolipids ...

  14. Characterization of biofilm formation by Borrelia burgdorferi in vitro.

    Directory of Open Access Journals (Sweden)

    Eva Sapi

    Full Text Available Borrelia burgdorferi, the causative agent of Lyme disease, has long been known to be capable of forming aggregates and colonies. It was recently demonstrated that Borrelia burgdorferi aggregate formation dramatically changes the in vitro response to hostile environments by this pathogen. In this study, we investigated the hypothesis that these aggregates are indeed biofilms, structures whose resistance to unfavorable conditions are well documented. We studied Borrelia burgdorferi for several known hallmark features of biofilm, including structural rearrangements in the aggregates, variations in development on various substrate matrices and secretion of a protective extracellular polymeric substance (EPS matrix using several modes of microscopic, cell and molecular biology techniques. The atomic force microscopic results provided evidence that multilevel rearrangements take place at different stages of aggregate development, producing a complex, continuously rearranging structure. Our results also demonstrated that Borrelia burgdorferi is capable of developing aggregates on different abiotic and biotic substrates, and is also capable of forming floating aggregates. Analyzing the extracellular substance of the aggregates for potential exopolysaccharides revealed the existence of both sulfated and non-sulfated/carboxylated substrates, predominately composed of an alginate with calcium and extracellular DNA present. In summary, we have found substantial evidence that Borrelia burgdorferi is capable of forming biofilm in vitro. Biofilm formation by Borrelia species might play an important role in their survival in diverse environmental conditions by providing refuge to individual cells.

  15. Isolation and characterization of Borrelia burgdorferi from Illinois Ixodes dammini.

    OpenAIRE

    Nelson, J A; Bouseman, J K; Kitron, U.; Callister, S M; Harrison, B; Bankowski, M. J; Peeples, M E; Newton, B.J.; Anderson, J F

    1991-01-01

    Ixodes dammini ticks from two northwestern Illinois sites were found to be infected with Borrelia burgdorferi at rates of 19 and 32%. B. burgdorferi isolates, one from each site, had protein and antigenic patterns similar to those of the B-31 strain. An indirect immunofluorescence method proved to be more sensitive than dark-field microscopy in detection of these spirochetes. A modified BSK medium containing rifampin was found to be more efficient for spirochete isolation than unsupplemented ...

  16. In Vitro Susceptibility Testing of Four Antibiotics against Borrelia burgdorferi: a Comparison of Results for the Three Genospecies Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi Sensu Stricto

    OpenAIRE

    Sicklinger, Martin; Wienecke, Ralf; Neubert, Uwe

    2003-01-01

    MICs and minimal bactericidal concentrations (MBCs) were evaluated for the four antibiotics azithromycin, amoxicillin, ceftriaxone, and doxycycline against the three main genospecies of Borrelia burgdorferi sensu lato. In MBC testing, statistically significant differences between the genospecies could be found in 7 out of 12 comparative evaluations (P < 0.05).

  17. Whole-Genome Sequences of Thirteen Isolates of Borrelia burgdorferi

    Energy Technology Data Exchange (ETDEWEB)

    Schutzer S. E.; Dunn J.; Fraser-Liggett, C. M.; Casjens, S. R.; Qiu, W.-G.; Mongodin, E. F.; Luft, B. J.

    2011-02-01

    Borrelia burgdorferi is a causative agent of Lyme disease in North America and Eurasia. The first complete genome sequence of B. burgdorferi strain 31, available for more than a decade, has assisted research on the pathogenesis of Lyme disease. Because a single genome sequence is not sufficient to understand the relationship between genotypic and geographic variation and disease phenotype, we determined the whole-genome sequences of 13 additional B. burgdorferi isolates that span the range of natural variation. These sequences should allow improved understanding of pathogenesis and provide a foundation for novel detection, diagnosis, and prevention strategies.

  18. Distribution of Borrelia burgdorferi Sensu Lato in China▿†

    OpenAIRE

    Hao, Qin; Hou, Xuexia; Geng, Zhen; Wan, Kanglin

    2011-01-01

    We genotyped 102 Borrelia burgdorferi sensu lato strains isolated from ticks, animals, and patients in 11 provinces in China by PCR–restriction fragment length polymorphism (PCR-RFLP) amplification of 5S (rrf)-23S (rrl) rRNA gene spacer amplicons and multilocus sequence analysis (MLSA). The results showed that Borrelia garinii was the main genotype in China (65/102) and that it was distributed mainly in northern China. Borrelia afzelii was the second most frequently found species (22/102), an...

  19. Lack of Serum Antibodies against Borrelia burgdorferi in Children with Autism

    OpenAIRE

    Burbelo, Peter D.; Swedo, Susan E.; Thurm, Audrey; Bayat, Ahmad; Levin, Andrew E.; Marques, Adriana; Iadarola, Michael J

    2013-01-01

    It has been proposed that Borrelia burgdorferi infection is present in ∼25% of children with autism spectrum disorders. In this study, antibodies against Borrelia burgdorferi were assessed in autistic (n = 104), developmentally delayed (n = 24), and healthy control (n = 55) children. No seropositivity against Borrelia burgdorferi was detected in the children with and without autism. There was no evidence of an association between Lyme disease and autism.

  20. Immunogenic integral membrane proteins of Borrelia burgdorferi are lipoproteins.

    Science.gov (United States)

    Brandt, M E; Riley, B S; Radolf, J D; Norgard, M V

    1990-04-01

    The pathogenic spirochete Borrelia burgdorferi contains a set of integral membrane proteins which were selectively extracted into the detergent phase upon solubilization of intact B. burgdorferi with the nonionic detergent Triton X-114. Virtually all of these hydrophobic proteins were recognized by antibodies in pooled sera from patients with chronic Lyme arthritis, demonstrating that proteins partitioning into the detergent phase of Triton X-114 encompass the major B. burgdorferi immunogens. Furthermore, most of these immunogenic proteins, including the previously characterized OspA and OspB membrane antigens, could be biosynthetically labeled when B. burgdorferi was incubated in vitro with [3H]palmitate. The OspA and OspB antigens were radioimmunoprecipitated from [3H]palmitate-labeled detergent-phase proteins with monoclonal antibodies, and [3H]palmitate was recovered unaltered from these proteins after sequential alkaline and acid hydrolyses. The combined results provide formal confirmation that the major B. burgdorferi immunogens extracted by Triton X-114 are lipoproteins. The demonstration that B. burgdorferi integral membrane antigens are lipoproteins may explain the basis of their immunogenicity and may help to improve our understanding of the surface topology of B. burgdorferi. PMID:2318538

  1. Comparison of Growth of Borrelia afzelii, Borrelia garinii, and Borrelia burgdorferi Sensu Stricto at Five Different Temperatures

    Science.gov (United States)

    Veinović, Gorana; Ružić-Sabljić, Eva; Strle, Franc; Cerar, Tjaša

    2016-01-01

    Lyme borreliosis is caused by the spirochete Borrelia burgdorferi sensu lato, a fastidious bacterium that replicates slowly and requires special conditions to grow in the laboratory. Borrelia isolation from clinical material is a golden standard for microbiological diagnosis of borrelial infection. Important factors that affect in vitro borrelia growth are temperature of incubation and number of borrelia cells in the sample. The aim of the study was to assess the influence of temperature on borrelia growth and survival by evaluation and comparison of growth of 31 different borrelia strains at five different temperatures and to determine the influence of different inoculums on borrelia growth at different temperatures. Borreliae were cultured in the MKP medium; the initial and final number of spirochetes was determined by dark field microscopy using Neubauer counting chamber. The growth of borrelia was defined as final number of cells/mL after three days of incubation. For all three Borrelia species, the best growth was found at 33°C, followed by 37, 28, and 23°C, while no growth was detected at 4°C (Pburgdorferi sensu stricto at 28, 33, and 37°C (P>0.05), respectively. Inoculum had statistically significant influence on growth of all three Borrelia species at all tested temperatures except at 4°C. PMID:27310556

  2. Distribution of Borrelia burgdorferi in host mice in Pennsylvania.

    OpenAIRE

    Lord, R D; Lord, V R; Humphreys, J. G.; McLean, R G

    1994-01-01

    Host mice (Peromyscus leucopus and Peromyscus maniculatus) were sampled throughout the state of Pennsylvania to determine the geographical and ecological distribution of the Lyme disease spirochete Borrelia burgdorferi. All 67 counties of the state were sampled. A total of 1,619 mice were captured from a total of 157 sites during the period 1990 to 1993 for an overall capture rate of 29.69%. A total of 112 (6.92%) isolations of B. burgdorferi were made. The distribution of isolations revealed...

  3. Linear and Circular Plasmid Content in Borrelia burgdorferi Clinical Isolates

    OpenAIRE

    Iyer, Radha; Kalu, Ogori; Purser, Joye; Norris, Steven; Stevenson, Brian; Schwartz, Ira

    2003-01-01

    The genome of Borrelia burgdorferi, the etiologic agent of Lyme disease, is composed of a linear chromosome and more than 20 linear and circular plasmids. Typically, plasmid content analysis has been carried out by pulsed-field gel electrophoresis and confirmed by Southern hybridization. However, multiple plasmids of virtually identical sizes (e.g., lp28 and cp32) complicate the interpretation of such data. The present study was undertaken to investigate the complete plasmid complements of B....

  4. Minimal-Change Disease Secondary to Borrelia burgdorferi Infection

    OpenAIRE

    Ewa Kwiatkowska; Edyta Gołembiewska; Kazimierz Ciechanowski; Karolina Kędzierska

    2012-01-01

    Lyme borreliosis is a chronic illness caused by tick-transmitted spirochete Borrelia burgdorferi. Borreliosis can be extremely threatening if it is not diagnosed and treated in early stages. Kidneys are not typically involved in the disease. However, in infected dogs, Lyme nephritis is present in 5–10% of cases. It is associated with rapidly progressing renal failure. Histopathological examination shows mesangial proliferative glomerulonephritis with diffuse tubular necrosis, (Dambach et al. ...

  5. Proteome Analysis of Borrelia burgdorferi Response to Environmental Change

    Energy Technology Data Exchange (ETDEWEB)

    Angel, Thomas E.; Luft, Benjamin J.; Yang, Xiaohua; Nicora, Carrie D.; Camp, David G.; Jacobs, Jon M.; Smith, Richard D.

    2010-11-02

    We examined global changes in protein expression in the B31 strain of Borrelia burgdorferi, in response to two environmental cues (pH and temperature) chosen for their reported similarity to those encountered at different stages of the organism’s life cycle. Multidimensional nano-liquid chromatographic separations coupled with tandem mass spectrometry were used to examine the array of proteins (i.e., the proteome) of B. burgdorferi for different pH and temperature culture conditions. Changes in pH and temperature elicited in vitro adaptations of this spirochete known to cause Lyme disease and led to alterations in protein expression that are associated with increased microbial pathogenesis. We identified 1031 proteins that represent 59% of the annotated genome of B. burgdorferi and elucidated a core proteome of 414 proteins that were present in all environmental conditions investigated. Observed changes in protein abundances indicated varied replicon usage, as well as proteome functional distributions between the in vitro cell culture conditions. Surprisingly, the pH and temperature conditions that mimicked B. burgdorferi residing in the gut of a fed tick showed a marked reduction in protein diversity. Additionally, the results provide us with leading candidates for exploring how B. burgdorferi adapts to and is able to survive in a wide variety of environmental conditions and lay a foundation for planned in situ studies of B. burgdorferi isolated from the tick midgut and infected animals.

  6. Genetic Characterization of Four Strains Borrelia Burgdorferi Isolated in China

    Institute of Scientific and Technical Information of China (English)

    曾霞; 王树声; 张涛; 毕胜利; 周永东

    2004-01-01

    To study the genetic characterization of four strains of Borrelia burgdorferi isolated in China. PCR technique was used to amplify the 5S-23S rRNA intergenic spacer DNA from the whole cellular DNA of isolated GXLD-4, 9, 18 and Chang 14, and then the amplified products were cloned into plasmid pGEM-T Easy and sequenced. It was found that the 5S-23S rRNA intergenic spacer DNA of the four isolates was 242 bp, revealing the nucleotide sequence identity of more than 99%. The four isolates had higher sequence identify with Borrelia valaisiana than with other genetic groups. These four isolates most likely belong to Borrelia valaisiana genomic group.

  7. Seroprevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, Ehrlichia canis, and Dirofilaria immitis among dogs in Canada

    OpenAIRE

    Villeneuve, Alain; Goring, Jonas; Marcotte, Lynne; Overvelde, Sébastien

    2011-01-01

    The seropositivity of dogs to Borrelia burgdorferi, Anaplasma phagocytophilum, and Ehrlichia canis antibodies, and Dirofilaria immitis antigen was assessed in Canada. Borrelia burgdorferi had the highest seroprevalence, while that of Dirofilaria immitis has not changed significantly in the past 20 y. The risk for these vector-borne infectious agents in Canadian dogs is low but widespread with foci of higher prevalence.

  8. Whole genome sequence of an unusual Borrelia burgdorferi sensu lato isolate

    Energy Technology Data Exchange (ETDEWEB)

    Casjens, S.R.; Dunn, J.; Fraser-Liggett, C. M.; Mongodin, E. F.; Qiu, W. G.; Luft, B. J.; Schutzer, S. E.

    2011-03-01

    Human Lyme disease is caused by a number of related Borrelia burgdorferi sensu lato species. We report here the complete genome sequence of Borrelia sp. isolate SV1 from Finland. This isolate is to date the closest known relative of B. burgdorferi sensu stricto, but it is sufficiently genetically distinct from that species that it and its close relatives warrant its candidacy for new-species status. We suggest that this isolate should be named 'Borrelia finlandensis.'

  9. Lipid exchange between Borrelia burgdorferi and host cells.

    Directory of Open Access Journals (Sweden)

    Jameson T Crowley

    2013-01-01

    Full Text Available Borrelia burgdorferi, the agent of Lyme disease, has cholesterol and cholesterol-glycolipids that are essential for bacterial fitness, are antigenic, and could be important in mediating interactions with cells of the eukaryotic host. We show that the spirochetes can acquire cholesterol from plasma membranes of epithelial cells. In addition, through fluorescent and confocal microscopy combined with biochemical approaches, we demonstrated that B. burgdorferi labeled with the fluorescent cholesterol analog BODIPY-cholesterol or (3H-labeled cholesterol transfer both cholesterol and cholesterol-glycolipids to HeLa cells. The transfer occurs through two different mechanisms, by direct contact between the bacteria and eukaryotic cell and/or through release of outer membrane vesicles. Thus, two-way lipid exchange between spirochetes and host cells can occur. This lipid exchange could be an important process that contributes to the pathogenesis of Lyme disease.

  10. Regulation of Expression of the Paralogous Mlp Family in Borrelia burgdorferi

    OpenAIRE

    Yang, Xiaofeng F.; Hübner, Anette; Popova, Taissia G.; Hagman, Kayla E.; Norgard, Michael V.

    2003-01-01

    The Mlp (multicopy lipoproteins) family is one of many paralogous protein families in Borrelia burgdorferi. To examine the extent to which the 10 members of the Mlp family in B. burgdorferi strain 297 might be differentially regulated, antibodies specific for each of the Mlps were developed and used to analyze the protein expression profiles of individual Mlps when B. burgdorferi replicated under various cultivation conditions. All of the Mlps were upregulated coordinately when B. burgdorferi...

  11. DNA Sequencing Diagnosis of Off-Season Spirochetemia with Low Bacterial Density in Borrelia burgdorferi and Borrelia miyamotoi Infections

    OpenAIRE

    Sin Hang Lee; Vigliotti, Jessica S.; Vigliotti, Veronica S; William Jones; Moorcroft, Thomas A.; Katherine Lantsman

    2014-01-01

    A highly conserved 357-bp segment of the 16S ribosomal RNA gene (16S rDNA) of Borrelia burgdorferi sensu lato and the correspondent 358-bp segment of the Borrelia miyamotoi gene were amplified by a single pair of nested polymerase chain reaction (PCR) primers for detection, and the amplicons were used as the templates for direct Sanger DNA sequencing. Reliable molecular diagnosis of these borreliae was confirmed by sequence alignment analysis of the hypervariable regions of the PCR amplicon,...

  12. Acquisition of Coinfection and Simultaneous Transmission of Borrelia burgdorferi and Ehrlichia phagocytophila by Ixodes scapularis Ticks

    OpenAIRE

    Michael L Levin; Fish, Durland

    2000-01-01

    The agents of Lyme disease (Borrelia burgdorferi) and human granulocytic ehrlichiosis (Ehrlichia phagocytophila) are both transmitted by the tick Ixodes scapularis. In nature, ticks are often infected with both agents simultaneously. We studied whether previous infection with either Borrelia or Ehrlichia in ticks would affect acquisition and transmission of a second pathogen. Ehrlichia-infected I. scapularis nymphs were fed upon Borrelia-infected mice, and Borrelia-infected I. scapularis nymp...

  13. Rapid Typing of Borrelia burgdorferi Sensu Lato Species in Specimens from Patients with Different Manifestations of Lyme Borreliosis

    OpenAIRE

    Lünemann, Jan D.; Zarmas, Silvia; Priem, Susanne; Franz, Juliane; Zschenderlein, Rolf; Aberer, Elisabeth; Klein, Rolf; Schouls, Leon; Burmester, Gerd R.; Krause, Andreas

    2001-01-01

    To further investigate the pathogenic potential of different Borrelia burgdorferi genospecies, specimens from 27 patients with different manifestations of Lyme borreliosis were analyzed by PCR and reverse line blotting (RLB). In samples from Lyme arthritis patients, B. burgdorferi sensu stricto was predominantly identified, while in patients with neuroborreliosis or acrodermatitis, Borrelia garinii and Borrelia afzelii, respectively, were exclusively detected. The results demonstrate that PCR...

  14. Population dynamics of Borrelia burgdorferi in Lyme disease

    Directory of Open Access Journals (Sweden)

    SebastianChristophBinder

    2012-03-01

    Full Text Available Many chronic inflammatory diseases are known to be caused by persistent bacterial or viral infections. A well-studied example is the tick-borne infection by the gram-negative Spirochaetes of the genus Borrelia in humans and other mammals, causing severe symptoms of chronic inflammation and subsequent tissue damage (Lyme Disease, particularly in large joints and the central nervous system, but also in the heart and other tissues of untreated patients. Although killed efficiently by human phagocytic cells in vitro, Borrelia exhibits a remarkably high infectivity in mice and men. In experimentally infected mice, the first immune response almost clears the infection. However, approximately one week post infection, the bacterial population recovers and reaches an even larger size before entering the chronic phase. We developed a mathematical model describing the bacterial growth and the immune response against Borrelia burgdorferi in the C3H mouse strain that has been established as an experimental model for Lyme disease. The peculiar dynamics of the infection exclude two possible mechanistic explanations for the regrowth of the almost cleared bacteria. Neither the hypothesis of bacterial dissemination to different tissue nor a limitation of phagocytic capacity were compatible with experiment. The mathematical model predicts that Borrelia recovers from the strong initial immune response by the regrowth of an immune-resistant sub-population of the bacteria. The chronic phase appears as an equilibration of bacterial growth and adaptive immunity. This result has major implications for the development of the chronic phase of Borrelia infections as well as on potential protective clinical interventions.

  15. Pleomorphic forms of Borrelia burgdorferi induce distinct immune responses.

    Science.gov (United States)

    Meriläinen, Leena; Brander, Heini; Herranen, Anni; Schwarzbach, Armin; Gilbert, Leona

    2016-01-01

    Borrelia burgdorferi is the causative agent of tick-borne Lyme disease. As a response to environmental stress B. burgdorferi can change its morphology to a round body form. The role of B. burgdorferi pleomorphic forms in Lyme disease pathogenesis has long been debated and unclear. Here, we demonstrated that round bodies were processed differently in differentiated macrophages, consequently inducing distinct immune responses compared to spirochetes in vitro. Colocalization analysis indicated that the F-actin participates in internalization of both forms. However, round bodies end up less in macrophage lysosomes than spirochetes suggesting that there are differences in processing of these forms in phagocytic cells. Furthermore, round bodies stimulated distinct cytokine and chemokine production in these cells. We confirmed that spirochetes and round bodies present different protein profiles and antigenicity. In a Western blot analysis Lyme disease patients had more intense responses to round bodies when compared to spirochetes. These results suggest that round bodies have a role in Lyme disease pathogenesis. PMID:27139815

  16. Conservation and Heterogeneity of vlsE among Human and Tick Isolates of Borrelia burgdorferi

    OpenAIRE

    Iyer, Radha; Hardham, John M.; Wormser, Gary P.; Schwartz, Ira; Norris, Steven J

    2000-01-01

    The vls (variable major protein [VMP]-like sequence) locus of Borrelia burgdorferi encodes an antigenic variation system that closely resembles the VMP system of relapsing fever borreliae. To determine whether vls sequences are present consistently in low-passage, infectious isolates of B. burgdorferi, 22 blood and erythema migrans biopsy isolates from Lyme disease patients in Westchester County, New York, were examined by Southern blot and PCR analysis. Each of the strains contained a single...

  17. Molecular characterization of Borrelia burgdorferi sensu lato strains isolated in the area of Belgrade, Serbia Caracterização molecular de cepas de Borrelia burgdorferi sensu lato isoladas na região de Belgrado, Sérvia

    OpenAIRE

    Elizabeta S. Ristanovic; Kayoko Kitamura; Toshiyuki Masuzawa; Marija J. Milutinovic; Radovan M. Cekanac; Novica T. Stajkovic; Danijela M. Zivanovic

    2007-01-01

    This is the first report of the molecular characterization and identification of Borrelia burgdorferi sensu lato strains isolated in Serbia. Isolates A1, A2 and M1, from Ixodes ricinus, belong to Borrelia burgdorferi sensu stricto, while isolate K1 from Apodemus flavicollis is a mixture of Borrelia afzelii and B. burgdorferi s.s.Trata-se do primeiro relato de identificação e caracterização molecular de cepas de Borrelia burgdorferi sensu lato isoladas na região de Belgrado, Sérvia. As cepas A...

  18. Ability to cause erythema migrans differs between Borrelia burgdorferi sensu lato isolates

    NARCIS (Netherlands)

    E. Tijsse-Klasen (Ellen); N. Pandak (Nenad); P. Hengeveld (Paul); K. Takumi (Katsuhisa); M.P.G. Koopmans D.V.M. (Marion); H. Sprong (Hein)

    2013-01-01

    textabstractBackground: Lyme borreliosis is a tick-borne disease caused by Borrelia burgdorferi sensu lato. The variety of characteristic and non-specific clinical manifestations is partially explained by its genetic diversity. We investigated the ability of B. burgdorferi sl isolates to cause eryth

  19. Evaluation of a quantitative fluorescence immunoassay (FIAX) for detection of serum antibody to Borrelia burgdorferi.

    OpenAIRE

    Pennell, D R; Wand, P J; Schell, R F

    1987-01-01

    A quantitative, indirect, fluorescence immunoassay (FIAX; Whittaker Bioproducts, Inc.) was compared with the conventional indirect fluorescent-antibody test for detection of serum antibody to Borrelia burgdorferi. FIAX correlated well with the indirect fluorescent-antibody test (r = 0.72). FIAX is a convenient and dependable means of measuring serum antibody to B. burgdorferi.

  20. Simultaneous transmission of Borrelia burgdorferi and Babesia microti by individual nymphal Ixodes dammini ticks.

    OpenAIRE

    Piesman, J; Hicks, T.C.; Sinsky, R J; Obiri, G.

    1987-01-01

    Nymphal Ixodes dammini ticks, selected from a group of ticks in which 22 of 31 (71%) contained dual Borrelia burgdorferi and Babesia microti infections, simultaneously transmitted B. burgdorferi and B. microti to 4 of 7 (57%) hamsters exposed to individual ticks.

  1. Characterization of Borrelia burgdorferi isolates by restriction endonuclease analysis and DNA hybridization.

    OpenAIRE

    LeFebvre, R B; Perng, G C; Johnson, R C

    1989-01-01

    Genomes of several Borrelia burgdorferi isolates from North America and Europe were characterized by restriction endonuclease analysis and DNA hybridization using labeled B. burgdorferi whole-cell DNA (strain ATCC 35210). Several different restriction and homology patterns were observed among these isolates, indicating genotypic heterogeneity within this genus and species. It was concluded from this study that restriction endonuclease analysis of B. burgdorferi whole-cell DNA may be a reliabl...

  2. The critical role of the linear plasmid lp36 in the infectious cycle of Borrelia burgdorferi

    OpenAIRE

    Mollie W Jewett; Lawrence, Kevin; Bestor, Aaron C; Tilly, Kit; Grimm, Dorothee; Shaw, Pamela; VanRaden, Mark; Gherardini, Frank; Rosa, Patricia A.

    2007-01-01

    Borrelia burgdorferi, the aetiological agent of Lyme disease, follows a life cycle that involves passage between the tick vector and the mammalian host. To investigate the role of the 36 kb linear plasmid, lp36 (also designated the B. burgdorferi K plasmid), in the infectious cycle of B. burgdorferi, we examined a clone lacking this plasmid, but containing all other plasmids known to be required for infectivity. Our results indicated that lp36 was not required for spirochete survival in the t...

  3. Evidence for Host-Genotype Associations of Borrelia burgdorferi Sensu Stricto

    OpenAIRE

    Samir Mechai; Gabriele Margos; Feil, Edward J.; Nicole Barairo; L Robbin Lindsay; Pascal Michel; Nicholas H Ogden

    2016-01-01

    Different genotypes of the agent of Lyme disease in North America, Borrelia burgdorferi sensu stricto, show varying degrees of pathogenicity in humans. This variation in pathogenicity correlates with phylogeny and we have hypothesized that the different phylogenetic lineages in North America reflect adaptation to different host species. In this study, evidence for host species associations of B. burgdorferi genotypes was investigated using 41 B. burgdorferi-positive samples from five mammal s...

  4. The urokinase receptor (uPAR) facilitates clearance of Borrelia burgdorferi

    NARCIS (Netherlands)

    J.W.R. Hovius; M.F. Bijlsma; G.J.W. van der Windt; W.J. Wiersinga; B.J.D. Boukens; J. Coumou; A. Oei; R. de Beer; A.F. de Vos; C. van 't Veer; A.P. van Dam; P. Wang; E. Fikrig; M.M. Levi; J.J.T.H. Roelofs; T. van der Poll

    2009-01-01

    The causative agent of Lyme borreliosis, the spirochete Borrelia burgdorferi, has been shown to induce expression of the urokinase receptor (uPAR); however, the role of uPAR in the immune response against Borrelia has never been investigated. uPAR not only acts as a proteinase receptor, but can also

  5. Borrelia burgdorferi sensu lato in humans in a rural area of Paraná State, Brazil

    OpenAIRE

    Daniela Dib Gonçalves; Rodrigo Assunção Moura; Mônica Nunes; Teresa Carreira; Odilon Vidotto; Julio Cesar de Freitas; Maria Luísa Vieira

    2015-01-01

    This study describes the detection of Borrelia garinii and Borrelia burgdorferi sensu stricto (s.s.) in Brazilian individuals using PCR and DNA sequencing. Our results suggest that these species are emerging pathogens in this country, and additional studies are necessary to determine the epidemiological characteristics of this disease in Brazil.

  6. Borrelia burgdorferi sensu lato in humans in a rural area of Paraná State, Brazil

    Directory of Open Access Journals (Sweden)

    Daniela Dib Gonçalves

    2015-06-01

    Full Text Available This study describes the detection of Borrelia garinii and Borrelia burgdorferi sensu stricto (s.s. in Brazilian individuals using PCR and DNA sequencing. Our results suggest that these species are emerging pathogens in this country, and additional studies are necessary to determine the epidemiological characteristics of this disease in Brazil.

  7. Occurrence of multiple infections with different Borrelia burgdorferi genospecies in Danish Ixodes ricinus nymphs

    DEFF Research Database (Denmark)

    Andersen, Jean Vennestrøm; Egholm, H.; Mikkelsen, Per Jensen

    2008-01-01

    The pathogen Borrelia burgdorferi causes Lyme Borreliosis in human and animals world-wide. In Europe the pathogen is transmitted to the host by the vector Ixodes ricinus. The nymph is the primary instar for transmission to humans. We here study the infection rate of five Borrelia genospecies: B...

  8. Common Ancestry of Borrelia burgdorferi Sensu Lato Strains from North America and Europe

    OpenAIRE

    Postic, D; Ras, N. Marti; Lane, R S; Humair, P.-F.; Wittenbrink, M. M.; Baranton, G

    1999-01-01

    Ten atypical European Borrelia burgdorferi sensu lato (Borrelia spp.) strains were genetically characterized, and the diversity was compared to that encountered among related Borrelia spp. from North America. Phylogenetic analyses of a limited region of the genome and of the whole genome extend existing knowledge about borrelial diversity reported earlier in Europe and the United States. Our results accord with the evidence that North American and European strains may have a common ancestry.

  9. Infection of Ixodes ricinus (Acari: Ixodidae) by Borrelia burgdorferi sensu lato in North Africa

    Science.gov (United States)

    Zhioua, E.; Bouattour, A.; Hu, C.M.; Gharbi, M.; Aeschliman, A.; Ginsberg, H.S.; Gern, L.

    1999-01-01

    Free-living adult Ixodes ricinus L. were collected in Amdoun, situated in the Kroumiry mountains in northwestern Tunisia (North Africa). Using direct fluorescence antibody assay, the infection rate of field-collected I. ricinus by Borrelia burgdorferi sensu lato was 30.5% (n = 72). No difference in infection rate was observed between male and female ticks. Spirochetes that had been isolated from I. ricinus from Ain Drahim (Kroumiry Mountains) in 1988 were identified as Borrelia lusitaniae (formerly genospecies PotiB2). This is the first identification of a genospecies of Borrelia burgdorferi sensu lato from the continent of Africa.

  10. In-vitro-Sensibilität von Borrelia burgdorferi sensu lato gegenüber vier Antibiotika: ein Vergleich der Genospezies Borrelia afzelii, Borrelia garinii und Borrelia burgdorferi sensu stricto

    OpenAIRE

    Sicklinger, Martin

    2006-01-01

    MICs and minimal bactericidal concentrations (MBCs) were evaluated for the four antibiotics azithromycin, amoxicillin, ceftriaxone, and doxycycline against the three main genospecies of Borrelia burgdorferi sensu lato. In MBC testing, statistically significant differences between the genospecies could be found in 7 out of 12 comparative evaluations (P < 0.05).

  11. Isolation, cultivation, and in vitro susceptibility testing of Borrelia burgdorferi sensu lato: A review

    Directory of Open Access Journals (Sweden)

    Veinović Gorana

    2013-01-01

    Full Text Available Lyme borreliosis is the most common vector-borne disease in the northern hemisphere. The agents of Lyme borreliosis are borrelia, bacteria of the family Spirochaetaceae, which are grouped in Borrelia burgdorferi sensu lato species complex. Borreliae are fastidious, slow-growing and biochemically inactive bacteria that need special attention and optimal conditions for cultivation. The isolation of Borrelia from clinical material and their cultivation is a time-consuming and demanding procedure. Cultivation lasts from 9 up to 12 weeks, which is much longer than is necessary to grow most other human bacterial pathogens. Although B. burgdorferi sensu lato is susceptible to a wide range of antimicrobial agents in vitro, up to now the susceptibility of individual Borrelia species to antibiotics is defined only partially. [Projekat Ministarstva nauke Republike Srbije, br. 175011

  12. Borrelia

    Czech Academy of Sciences Publication Activity Database

    Rudenko, Natalia; Golovchenko, Maryna; Oliver, J. H., Jr.; Grubhoffer, Libor

    Boca Raton: CRC Press, Taylor & Francis Group, 2011 - (Liu, D.), s. 1155-1168 ISBN 978-1-4398-1238-9 R&D Projects: GA MŠk(CZ) LC06009; GA ČR GA206/09/1782 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia * Ixodid ticks * Borrelia burgdorferi sensu lato coplex Subject RIV: EE - Microbiology, Virology

  13. Sensitivity of spirochetes from \\kur{Borrelia burgdorferi} sensu lato complex to human complement: infection potential of selected species

    OpenAIRE

    Tichá, Lucie

    2015-01-01

    Sensitivity of spirochetes from Borrelia burgdorferi sensu lato complex to serum complement of humans of different age and sex was analyzed. Complement-mediated Borrelia killing was observed in different combination of serum and selected Borrelia genospecies. The obtained results confirmed that age itself does not influence the sensitivity of human to Borrelia infection. However, the females seem to be more vulnerable to it. Each of ten tested Borrelia species was proved to be potentially inf...

  14. Fourier transform infrared spectroscopy of DNA from Borrelia burgdorferi sensu lato and Ixodes ricinus ticks

    Science.gov (United States)

    Muntean, Cristina M.; Stefan, Razvan; Bindea, Maria; Cozma, Vasile

    2013-06-01

    In this work we present a method for detection of motile and immotile Borrelia burgdorferi genomic DNA, in relation with infectious and noninfectious spirochetes. An FT-IR study of DNA isolated from B. burgdorferi sensu lato strains and from positive and negative Ixodes ricinus ticks, respectively, is reported. Motile bacterial cells from the species B. burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii were of interest. Also, FT-IR absorbance spectra of DNA from immotile spirochetes of B. burgdorferi sensu stricto, in the absence and presence of different antibiotics (doxycycline, erythromycin, gentamicin, penicillin V or phenoxymethylpenicillin, tetracycline, respectively) were investigated. FT-IR spectra, providing a high molecular structural information, have been analyzed in the wavenumber range 400-1800 cm-1. FT-IR signatures, spectroscopic band assignments and structural interpretations of these DNAs are reported. Spectral differences between FT-IR absorbances of DNAs from motile bacterial cells and immotile spirochetes, respectively, have been found. Particularly, alterations of the sugar-phosphate B-form chain in the case of DNA from Borrelia immotile cells, as compared with DNA from B. burgdorferi sensu lato motile cells have been observed. Based on this work, specific B. burgdorferi sensu lato and I. ricinus DNA-ligand interactions, respectively, might be further investigated using Fourier transform infrared spectroscopy.

  15. Diversity within Borrelia burgdorferi sensu lato genospecies in Switzerland by recA gene sequence

    OpenAIRE

    Casati, Simona; Bernasconi, Marco V.; Gern, Lise; Piffaretti, Jean-Claude

    2006-01-01

    A total of 874 Ixodes ricinus ticks were collected in Switzerland to investigate the genetic diversity of the Borrelia population. We integrated to the RT-PCR method the DNA sequence analysis of a 162-bp fragment of the recA gene. Five genospecies were detected: Borrelia afzelii, Borrelia burgdorferi s.s., Borrelia garinii, Borrelia valaisiana, and Borrelia lusitaniae. A heterogeneous distribution was observed within the B. burgdorferi s.l. genospecies. The most prevalent and diverse genospec...

  16. Detection of Borrelia burgdorferi sensu stricto and Borrelia garinii DNAs in patient with Hyperkeratosis lenticularis perstans (Flegel disease).

    Science.gov (United States)

    Schwarzova, Katarina; Kozub, Peter; Szep, Zoltan; Golovchenko, Marina; Rudenko, Natasha

    2016-09-01

    Determination of the causative agent of erythema-like skin lesions in case of nonspecific superficial perivascular dermatitis was supported by histological examination and led to the latter diagnosis of Hyperkeratosis lenticularis perstans (Flegel disease) in patient. The presence of antibodies against Borrelia burgdorferi in patient serum was confirmed by a routine ELISA method and verified by Western blot technique. Skin biopsy and blood specimens were analyzed by PCR and multilocus sequence analysis (MLSA). Western blot method revealed IgG antibody response against two specific antigens, 17 and 83 kDa proteins. The recombinant test detected IgG antibody response against p100 and p41 antigens. The sequence analysis of amplicons from the selected genomic loci obtained from skin biopsy and serum samples revealed the presence of two species from B. burgdorferi sensu lato complex as a co-infection in this patient-B. burgdorferi sensu stricto (s.s.) and Borrelia garinii. PMID:26769152

  17. Rodent species as natural reservoirs of Borrelia burgdorferi sensu lato in different habitats of Ixodes ricinus in The Netherlands

    NARCIS (Netherlands)

    Gassner, F.; Takken, W.; Plas, C.; Kastelein, P.; Hoetmer, A.J.; Holdinga, M.; Overbeek, van L.S.

    2013-01-01

    Rodents are natural reservoirs for human pathogenic spirochaetes of the Borrelia burgdorferi complex [B. burgdorferi sensu lato (s.l.)], and the pathogens are transmitted by Ixodes ricinus ticks to humans in The Netherlands. B. burgdorferi s.l. infection prevalence in questing ticks, rodents, and ti

  18. SERUM ANTIBODIES TO BORRELIA BURGDORFERI, ANAPLASMA PHAGOCYTOPHILUM, AND BABESIA MICROTI IN RECAPTURED WHITE-FOOTED MICE

    OpenAIRE

    Magnarelli, Louis A.; Williams, Scott C.; Norris, Steven J; Fikrig, Erol

    2013-01-01

    A mark-release-recapture study was conducted during 2007 through 2010 in six, tick-infested sites in Connecticut, United States to measure changes in antibody titers for Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum, and Babesia microti in Peromyscus leucopus (white-footed mice). There was an overall recapture rate of 40%, but only four tagged mice were caught in ≥2 yr. Sera from 561 mice were analyzed for total antibodies to B. burgdorferi and A. phagocytophilum by using whol...

  19. Borrelia burgdorferi aggrecanase activity: more evidence for persistent infection in Lyme disease

    OpenAIRE

    Stricker, Raphael B.; Johnson, Lorraine

    2013-01-01

    Lyme disease is the most common tickborne illness in the world today. A recent study describes for the first time an enzyme produced by the spirochetal agent of Lyme disease, Borrelia burgdorferi, that cleaves aggrecan, a proteoglycan found in joints and connective tissue. Discovery of the spirochetal aggrecanase raises many questions about the pathogenesis of Lyme arthritis and lends support to the concept of persistent B. burgdorferi infection in patients with chronic Lyme disease symptoms.

  20. Survey for Ixodes spp. and Borrelia burgdorferi in southeastern Wisconsin and northeastern Illinois.

    OpenAIRE

    Callister, S M; Nelson, J A; Schell, R F; Jobe, D A; Bautz, R; Agger, W A; Coggins, J

    1991-01-01

    Forested areas adjacent to Milwaukee, Wis., and Chicago, Ill., were investigated for rodents and ticks infected with Borrelia burgdorferi, the causative agent of Lyme disease. White-footed mice (Peromyscus leucopus or Peromyscus maniculatus), meadow voles (Microtus pennsylvanicus), and eastern chipmunks (Tamias striatus) were captured; and specimens from these animals were cultured for B. burgdorferi to define whether the midwestern Lyme disease area currently encompasses these large metropol...

  1. Flagellin and outer surface proteins from Borrelia burgdorferi are not glycosylated

    OpenAIRE

    ŠTĚRBA, Ján

    2012-01-01

    Glycosylation of four proteins from Borrelia burgdorferi s.s. was investigated ? flagellins FlaA, FlaB, and outer surface proteins OspA and OspB. Glycosylation of these four proteins was not proved by any of the used techniques. However, other glycan-staining positive proteins were present in the borrelia samples. These proteins were suggested to originate in the culture medium.

  2. DNA Sequencing Diagnosis of Off-Season Spirochetemia with Low Bacterial Density in Borrelia burgdorferi and Borrelia miyamotoi Infections

    Directory of Open Access Journals (Sweden)

    Sin Hang Lee

    2014-06-01

    Full Text Available A highly conserved 357-bp segment of the 16S ribosomal RNA gene (16S rDNA of Borrelia burgdorferi sensu lato and the correspondent 358-bp segment of the Borrelia miyamotoi gene were amplified by a single pair of nested polymerase chain reaction (PCR primers for detection, and the amplicons were used as the templates for direct Sanger DNA sequencing. Reliable molecular diagnosis of these borreliae was confirmed by sequence alignment analysis of the hypervariable regions of the PCR amplicon, using the Basic Local Alignment Search Tool (BLAST provided by the GenBank. This methodology can detect and confirm B. burgdorferi and B. miyamotoi in blood samples of patients with off-season spirochetemia of low bacterial density. We found four B. miyamotoi infections among 14 patients with spirochetemia, including one patient co-infected by both B. miyamotoi and B. burgdorferi in a winter month when human exposure to tick bites is very limited in the Northeast of the U.S.A. We conclude that sensitive and reliable tests for these two Borrelia species should be implemented in the microbiology laboratory of hospitals located in the disease-endemic areas, for timely diagnosis and appropriate treatment of the patients at an early stage of the infection to prevent potential tissue damages.

  3. In vitro susceptibilities of Borrelia burgdorferi to five oral cephalosporins and ceftriaxone.

    OpenAIRE

    Agger, W A; Callister, S M; Jobe, D A

    1992-01-01

    We determined the in vitro susceptibilities of eight Borrelia burgdorferi isolates to five oral cephalosporins. MICs for B. burgdorferi 297 were 23 micrograms/ml (cephalexin), 45 micrograms/ml (cefadroxil), 91 micrograms/ml (cefaclor), 0.13 microgram/ml (cefuroxime), 0.8 microgram/ml (cefixime), and 0.02 microgram/ml (ceftriaxone). When B. burgdorferi isolates were exposed to concentrations twice the MIC of cefuroxime, cefixime, or ceftriaxone, at least 72 h of incubation was required to kill...

  4. Reservoir competence of Microtus pennsylvanicus (Rodentia: Cricetidae) for the Lyme disease spirochete, Borrelia burgdorferi

    Science.gov (United States)

    Markowski, D.; Ginsberg, H.S.; Hyland, K.E.; Hu, R.

    1998-01-01

    The reservoir competence of the meadow vole, Microtus pennsylvanicus Ord, for the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner was established on Patience Island, RI. Meadow voles were collected from 5 locations throughout Rhode Island. At 4 of the field sites, M. pennsylvanicus represented only 4.0% (n = 141) of the animals captured. However, on Patience Island, M. pennsylvanicus was the sole small mammal collected (n = 48). Of the larval Ixodes scapularis Say obtained from the meadow voles on Patience Island, 62% (n = 78) was infected with B. burgdorferi. Meadow voles from all 5 locations were successfully infected with B. burgdorferi in the laboratory and were capable of passing the infection to xenodiagnostic I. scapularis larvae for 9 wk. We concluded that M. pennsylvanicus was physiologically capable of maintaining B. burgdorferi infection. However, in locations where Peromyscus leucopus (Rafinesque) is abundant, the role of M. pennsylvanicus as a primary reservoir for B. burgdorferi was reduced.

  5. Suppression of Long-Lived Humoral Immunity Following Borrelia burgdorferi Infection.

    Directory of Open Access Journals (Sweden)

    Rebecca A Elsner

    2015-07-01

    Full Text Available Lyme Disease caused by infection with Borrelia burgdorferi is an emerging infectious disease and already by far the most common vector-borne disease in the U.S. Similar to many other infections, infection with B. burgdorferi results in strong antibody response induction, which can be used clinically as a diagnostic measure of prior exposure. However, clinical studies have shown a sometimes-precipitous decline of such antibodies shortly following antibiotic treatment, revealing a potential deficit in the host's ability to induce and/or maintain long-term protective antibodies. This is further supported by reports of frequent repeat infections with B. burgdorferi in endemic areas. The mechanisms underlying such a lack of long-term humoral immunity, however, remain unknown. We show here that B. burgdorferi infected mice show a similar rapid disappearance of Borrelia-specific antibodies after infection and subsequent antibiotic treatment. This failure was associated with development of only short-lived germinal centers, micro-anatomical locations from which long-lived immunity originates. These showed structural abnormalities and failed to induce memory B cells and long-lived plasma cells for months after the infection, rendering the mice susceptible to reinfection with the same strain of B. burgdorferi. The inability to induce long-lived immune responses was not due to the particular nature of the immunogenic antigens of B. burgdorferi, as antibodies to both T-dependent and T-independent Borrelia antigens lacked longevity and B cell memory induction. Furthermore, influenza immunization administered at the time of Borrelia infection also failed to induce robust antibody responses, dramatically reducing the protective antiviral capacity of the humoral response. Collectively, these studies show that B. burgdorferi-infection results in targeted and temporary immunosuppression of the host and bring new insight into the mechanisms underlying the failure

  6. Role of outer membrane architecture in immune evasion by Treponema pallidum and Borrelia burgdorferi.

    Science.gov (United States)

    Radolf, J D

    1994-09-01

    Combined ultrastructural and molecular studies have revealed that the syphilis and Lyme-disease spirochetes, Treponema pallidum and Borrelia burgdorferi, have distinctive molecular architectures. Both organisms persist in their hosts and have strategies for immune evasion that include the use of rare, poorly immunogenic surface-exposed proteins as potential virulence determinants. PMID:7812663

  7. Antibodies against Borrelia burgdorferi sensu lato among Adults, Germany, 2008–2011

    OpenAIRE

    Wilking, Hendrik; Fingerle, Volker; Klier, Christiane; Thamm, Michael; Stark, Klaus

    2015-01-01

    To assess Borrelia burgdorferi sensu lato (the cause of Lyme borreliosis) seropositivity in Germany, we tested serum samples from health survey (2008–2011) participants. Seroprevalence was 5.8% among women and 13.0% among men; infection risk was highest among persons >60 years of age. Public health interventions, including education about risk factors and preventive measures, are needed.

  8. Concurrent Borrelia burgdorferi and Babesia microti infection in nymphal Ixodes dammini.

    OpenAIRE

    Piesman, J; Mather, T N; Telford, S R; Spielman, A

    1986-01-01

    Ixodes dammini nymphs were examined for evidence of concurrent infection with Borrelia burgdorferi and Babesia microti. A total of 19 nymphs (18.6%) from Nantucket Island were simultaneously infected, as were 24 nymphs (8.2%) from Naushon Island. These observations are consistent with a common reservoir host for both I. dammini-transmitted pathogens.

  9. Lectin-binding characteristics of a Lyme borreliosis spirochete Borrelia burgdorferi sensu stricto

    Czech Academy of Sciences Publication Activity Database

    Vancová, M.; Nebesářová, J.; Grubhoffer, Libor

    2005-01-01

    Roč. 50, č. 3 (2005), s. 229-238. ISSN 0015-5632 R&D Projects: GA ČR GA206/03/1323; GA AV ČR KSK6005114 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia burgdorferi * electron microscopy * lectin binding Subject RIV: EE - Microbiology, Virology Impact factor: 0.918, year: 2005

  10. First isolation and cultivation of Borrelia burgdorferi sensu lato from Missouri.

    Science.gov (United States)

    Oliver, J H; Kollars, T M; Chandler, F W; James, A M; Masters, E J; Lane, R S; Huey, L O

    1998-01-01

    Five Borrelia burgdorferi sensu lato isolates from Missouri are described. This represents the first report and characterization of such isolates from that state. The isolates were obtained from either Ixodes dentatus or Amblyomma americanum ticks that had been feeding on cottontail rabbits (Sylvilagus floridanus) from a farm in Bollinger County, Mo., where a human case of Lyme disease had been reported. All isolates were screened immunologically by indirect immunofluorescence by using monoclonal antibodies to B. burgdorferi-specific outer surface protein A (OspA) (antibodies H3TS and H5332), B. burgdorferi-specific OspB (antibody H6831), Borrelia (genus)-specific antiflagellin (antibody H9724), and Borrelia hermsii-specific antibody (antibody H9826). Analysis of the isolates also involved a comparison of their protein profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Finally, the isolates were analyzed by PCR with six pairs of primers known to amplify selected DNA target sequences specifically found in the reference strain B. burgdorferi B-31. Although some genetic variability was detected among the five isolates as well as between them and the B-31 strain, enough similarities were found to classify them as B. burgdorferi sensu lato. PMID:9431909

  11. Complement-mediated killing of Borrelia burgdorferi by nonimmune sera from sika deer.

    Science.gov (United States)

    Nelson, D R; Rooney, S; Miller, N J; Mather, T N

    2000-12-01

    Various species of cervid deer are the preferred hosts for adult, black-legged ticks (Ixodes scapularis and Ixodes pacificus) in the United States. Although frequently exposed to the agent of Lyme disease (Borrelia burgdorferi), these animals, for the most part, are incompetent as transmission reservoirs. We examined the borreliacidal activity of normal and B. burgdorferi-immune sera from sika deer (Cervus nippon) maintained in a laboratory setting and compared it to that of similar sera from reservoir-competent mice and rabbits. All normal deer sera (NDS) tested killed > 90% of B. burgdorferi cells. In contrast, normal mouse and rabbit sera killed 1:1,024. Heat treatment (56 C, 30 min) of NDS reduced borreliacidal activity, with 90% of the cells killed. Addition of 10 mM EDTA reduced the killing to approximately 30%, whereas the addition of Mg2+ (10 mM) restored borreliacidal activity to NDS. The addition of zymosan A, an activator of the alternative pathway, increased the survival of B. burgdorferi cells to approximately 80% in NDS. These data suggest that the alternative complement activation pathway plays a major role in the borreliacidal activity of NDS. Additionally, 10 mM EGTA had almost no effect on the killing activity of B. burgdorferi-exposed deer sera, suggesting that the classical pathway is not involved in Borrelia killing, even in sera from B. burgdorferi-exposed deer. PMID:11191897

  12. The cyclic-di-GMP signaling pathway in the Lyme disease spirochete, Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Elizabeth A. Novak

    2014-05-01

    Full Text Available In nature, the Lyme disease spirochete Borrelia burgdorferi cycles between the unrelated environments of the Ixodes tick vector and mammalian host. In order to survive transmission between hosts, B. burgdorferi must be able to not only detect changes in its environment, but also rapidly and appropriately respond to these changes. One manner in which this obligate parasite regulates and adapts to its changing environment is through cyclic-di-GMP (c-di-GMP signaling. c-di-GMP has been shown to be instrumental in orchestrating the adaptation of B. burgdorferi to the tick environment. B. burgdorferi possesses only one set of c-di-GMP-metabolizing genes (one diguanylate cyclase and two distinct phosphodiesterases and one c-di-GMP-binding PilZ-domain protein designated as PlzA. While studies in the realm of c-di-GMP signaling in B. burgdorferi have exploded in the last few years, there are still many more questions than answers. Elucidation of the importance of c-di-GMP signaling to B. burgdorferi may lead to the identification of mechanisms that are critical for the survival of B. burgdorferi in the tick phase of the enzootic cycle as well as potentially delineate a role (if any c-di-GMP may play in the transmission and virulence of B. burgdorferi during the enzootic cycle, thereby enabling the development of effective drugs for the prevention and/or treatment of Lyme disease.

  13. Symptomatic co-infection with Babesia microti and Borrelia burgdorferi in patient after international exposure; a challenging case in Poland.

    Science.gov (United States)

    Jabłońska, Joanna; Żarnowska-Prymek, Hanna; Stańczak, Joanna; Kozłowska, Joanna; Wiercińska-Drapało, Alicja

    2016-06-01

    The report presents a well-documented case of symptomatic co-infection of Babesia microti and Borrelia burgdorferi in a Polish immunocompetent patient after travelling to Canada and the USA. PMID:27294655

  14. Critical Analysis of Treatment Trials of Rhesus Macaques Infected with Borrelia burgdorferi Reveals Important Flaws in Experimental Design

    OpenAIRE

    Wormser, Gary P.; Baker, Phillip J.; O'Connell, Susan; Pachner, Andrew R.; Schwartz, Ira; Shapiro, Eugene D.

    2012-01-01

    A critical analysis of two treatment trials of Chinese rhesus macaques infected with Borrelia burgdorferi indicates that insufficient attention was placed on documenting the blood levels, pharmacokinetics, and pharmacodynamic parameters of the antibiotics used in this host. Consequently, it is impossible to conclude that the findings have validity in judging the efficacy of doxycycline or ceftriaxone for the treatment of Borrelia burgdorferi in this animal model.

  15. Identification of glycoproteins in samples of the CB53 isolate of the spirochete \\kur{Borrelia burgdorferi} s.s.

    OpenAIRE

    JONÁKOVÁ, Martina

    2009-01-01

    Glycosylation of Borrelia burgdorferi has been studied for a quite long time, but it's not clear so far, if borrelia spirochetas are able to glycosylate proteins. This work is focused on isolation of glycoproteins from B. burgdorferi outer membranes and periplasmic flagella, on their identification and characterization. My results imply that OspC, FlaB and p83/100 proteins are potentially glycosylated.

  16. Effectiveness of Stevia Rebaudiana Whole Leaf Extract Against the Various Morphological Forms of Borrelia Burgdorferi in Vitro

    OpenAIRE

    Theophilus, P. A. S.; Victoria, M. J.; Socarras, K. M.; Filush, K. R.; Gupta, K.; Luecke, D. F.; Sapi, E.

    2015-01-01

    Lyme disease is a tick-borne multisystemic disease caused by Borrelia burgdorferi. Administering antibiotics is the primary treatment for this disease; however, relapse often occurs when antibiotic treatment is discontinued. The reason for relapse remains unknown, but recent studies suggested the possibilities of the presence of antibiotic resistant Borrelia persister cells and biofilms. In this study, we evaluated the effectiveness of whole leaf Stevia extract against B. burgdorferi spiroche...

  17. Polymerase chain reaction in diagnosis of Borrelia burgdorferi infections and studies on taxonomic classification

    DEFF Research Database (Denmark)

    Lebech, Anne-Mette

    2002-01-01

    Lyme borreliosis caused by the spirochete Borrelia burgdorferi is now the most common vectorborne disease in North America, Europe and Asia. It is a multisystemic infection which may cause skin, neurological, cardiac or rheumatologic disorders. The aims of the present thesis were: (i) to develop a...... PCR assay for direct detection of B. burgdorferi DNA and to evaluate the diagnostic utility of PCR in clinical specimens from patients with Lyme borreliosis and (ii) to study the taxonomic classification of B. burgdorferi isolates and its implications for epidemiology and clinical presentation....... Laboratory diagnosis of Lyme borreliosis by direct demonstration of B. burgdorferi in clinical specimens would compared to current serology allow (i) optimal specificity, (ii) increased sensitivity during the first weeks of infection, when the antibody response is not yet detectable and (iii) discrimination...

  18. Few vertebrate species dominate the Borrelia burgdorferi s.l. life cycle

    Science.gov (United States)

    Hofmeester, T. R.; Coipan, E. C.; van Wieren, S. E.; Prins, H. H. T.; Takken, W.; Sprong, H.

    2016-04-01

    Background. In the northern hemisphere, ticks of the Ixodidae family are vectors of diseases such as Lyme borreliosis, Rocky Mountain spotted fever and tick-borne encephalitis. Most of these ticks are generalists and have a three-host life cycle for which they are dependent on three different hosts for their blood meal. Finding out which host species contribute most in maintaining ticks and the pathogens they transmit, is imperative in understanding the drivers behind the dynamics of a disease. Methods. We performed a systematic review to identify the most important vertebrate host species for Ixodes ricinus and Borrelia burgdorferi s.l. as a well-studied model system for tick-borne diseases. We analyzed data from 66 publications and quantified the relative contribution for 15 host species. Review results. We found a positive correlation between host body mass and tick burdens for the different stages of I. ricinus. We show that nymphal burdens of host species are positively correlated with infection prevalence with B. burgdorferi s.l., which is again positively correlated with the realized reservoir competence of a host species for B. burgdorferi s.l. Our quantification method suggests that only a few host species, which are amongst the most widespread species in the environment (rodents, thrushes and deer), feed the majority of I. ricinus individuals and that rodents infect the majority of I. ricinus larvae with B. burgdorferi s.l. Discussion. We argue that small mammal-transmitted Borrelia spp. are maintained due to the high density of their reservoir hosts, while bird-transmitted Borrelia spp. are maintained due to the high infection prevalence of their reservoir hosts. Our findings suggest that Ixodes ricinus and Borrelia burgdorferi s.l. populations are maintained by a few widespread host species. The increase in distribution and abundance of these species, could be the cause for the increase in Lyme borreliosis incidence in Europe in recent decades.

  19. Study of the effect of \\kur{Borrelia burgdorferi} infection on the behaviour of Ixodes ricinus tick

    OpenAIRE

    HUSPEKOVÁ, Helena

    2014-01-01

    The main goal of this master thesis was to determine whether or not is the infection with Borrelia burgdorferi sensu lato responsible for changes in Ixodes ricinus tick behaviour. We tried to investigate if B. burgdorferi infection changes the time required for blood feeding, tick activity, phototaxis and amount of blood which is engorged by the tick.

  20. Identification of related DNA sequences in Borrelia burgdorferi and two strains of Leptospira interrogans by using polymerase chain reaction.

    OpenAIRE

    Kron, M A; Gupta, A; Mackenzie, C. D.

    1991-01-01

    The suitability of a polymerase chain reaction assay for Borrelia burgdorferi in epidemiological studies of infected tick populations was evaluated by using 28 strains of Leptospira interrogans and lysates of fixed adult Ixodes tick tissues. Two false positives representing leptospires were differentiated from B. burgdorferi by using an oligonucleotide probe.

  1. Large scale spatial risk and comparative prevalence of Borrelia miyamotoi and Borrelia burgdorferi sensu lato in Ixodes pacificus.

    Directory of Open Access Journals (Sweden)

    Kerry Padgett

    Full Text Available Borrelia miyamotoi is a newly described emerging pathogen transmitted to people by Ixodes species ticks and found in temperate regions of North America, Europe, and Asia. There is limited understanding of large scale entomological risk patterns of B. miyamotoi and of Borreila burgdorferi sensu stricto (ss, the agent of Lyme disease, in western North America. In this study, B. miyamotoi, a relapsing fever spirochete, was detected in adult (n=70 and nymphal (n=36 Ixodes pacificus ticks collected from 24 of 48 California counties that were surveyed over a 13 year period. Statewide prevalence of B. burgdorferi sensu lato (sl, which includes B. burgdorferi ss, and B. miyamotoi were similar in adult I. pacificus (0.6% and 0.8%, respectively. In contrast, the prevalence of B. burgdorferi sl was almost 2.5 times higher than B. miyamotoi in nymphal I. pacificus (3.2% versus 1.4%. These results suggest similar risk of exposure to B. burgdorferi sl and B. miyamotoi from adult I. pacificus tick bites in California, but a higher risk of contracting B. burgdorferi sl than B. miyamotoi from nymphal tick bites. While regional risk of exposure to these two spirochetes varies, the highest risk for both species is found in north and central coastal California and the Sierra Nevada foothill region, and the lowest risk is in southern California; nevertheless, tick-bite avoidance measures should be implemented in all regions of California. This is the first study to comprehensively evaluate entomologic risk for B. miyamotoi and B. burgdorferi for both adult and nymphal I. pacificus, an important human biting tick in western North America.

  2. Molecular Characterization of Borrelia burgdorferi Linear Plasmids by DNA Hybridization, PCR, Two-Dimensional Gel Electrophoresis, and Electron Microscopy

    OpenAIRE

    Karami, Ali; HOSSEYNI, Seyed Mohammad Javad; KIARUDI, Yaser

    2007-01-01

    Borrelia burgdorferi sensu stricto is the predominant cause of Lyme disease. B. burgdorferi strains carry several linear and circular plasmids and this may be also used as a distinguishing property of the bacteria. The presence of a mixed population of linear and circular plasmids makes separation and characterization of each kind very difficult. In this study, 7 clinical strains of skin and CSF isolates were determined by clonal selection and plasmid profile of B. burgdorferi by electron mic...

  3. Molecular Evidence of Coinfection of Ticks with Borrelia burgdorferi Sensu Lato and the Human Granulocytic Ehrlichiosis Agent in Switzerland

    OpenAIRE

    Leutenegger, Christian M.; Pusterla, Nicola; Mislin, Caroline N.; Weber, Rainer; Lutz, Hans

    1999-01-01

    Adult Ixodes ricinus ticks were collected in Switzerland and tested for the presence of coinfection with Borrelia burgdorferi sensu lato and the human granulocytic ehrlichiosis (HGE) agent by real-time PCR. Of 100 ticks, 49% were positive for B. burgdorferi and 2% were positive for the HGE agent. The two HGE agent-positive ticks were also found to be positive for B. burgdorferi.

  4. Identification of Borrelia burgdorferi genospecies isolated from Ixodes ricinus ticks in the South Moravian region of the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Bonczek, Ondřej; Žákovská, A.; Vargová, L.; Šerý, Omar

    2015-01-01

    Roč. 22, č. 4 (2015), s. 642-646. ISSN 1232-1966 Institutional support: RVO:67985904 Keywords : Borrelia burgdorferi s. I. * Borrelia spielmanii * Ixodes ricinus Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.126, year: 2014

  5. Seroprevalence of Borrelia burgdorferi antibodies in white-tailed deer from Texas

    OpenAIRE

    Adetunji, Shakirat A.; Rosina C. Krecek; Castellanos, Gabrielle; Morrill, John C.; Blue-McLendon, Alice; Walt E. Cook; Esteve-Gassent, Maria D.

    2016-01-01

    Lyme Disease is caused by the bacterial pathogen Borrelia burgdorferi, and is transmitted by the tick-vector Ixodes scapularis. It is the most prevalent arthropod-borne disease in the United States. To determine the seroprevalence of B. burgdorferi antibodies in white-tailed deer (Odocoileus virginianus) from Texas, we analyzed serum samples (n = 1493) collected during the 2001–2015 hunting seasons, using indirect ELISA. Samples with higher sero-reactivity (0.803 and above) than the negative ...

  6. Disparity in the natural cycles of Borrelia burgdorferi and the agent of human granulocytic ehrlichiosis.

    OpenAIRE

    Levin, M L; des Vignes, F.; Fish, D

    1999-01-01

    We studied the prevalence of Borrelia burgdorferi and the agent of human granulocytic ehrlichiosis (HGE) among questing nymphal and adult Ixodes scapularis ticks of the same generation and the infectivity of wild white-footed mice for ticks feeding on them. The prevalence of B. burgdorferi infection in host-seeking ticks increased less than twofold from nymphal (31% to 33%) to adult (52% to 56%) stage, and 52% of white-footed mice were infected. Prevalence of the agent of HGE increased 4.5- t...

  7. An Enhanced ELISPOT Assay for Sensitive Detection of Antigen-Specific T Cell Responses to Borrelia burgdorferi

    OpenAIRE

    Kellermann, Gottfried H.; Lehmann, Paul V; Diana R. Roen; Chenggang Jin

    2013-01-01

    Lyme Borreliosis is an infectious disease caused by the spirochete Borrelia burgdorferi that is transmitted through the bite of infected ticks. Both B cell-mediated humoral immunity and T cell immunity develop during natural Borrelia infection. However, compared with humoral immunity, the T cell response to Borrelia infection has not been well elucidated. In this study, a novel T cell-based assay was developed and validated for the sensitive detection of antigen-specific T cell response to B....

  8. Detection and differentiation of Borrelia burgdorferi sensu lato in ticks collected from sheep and cattle in China

    OpenAIRE

    Jorgensen Wayne; Ren Qiaoyun; Liu Aihong; Liu Junlong; Liu Zhijie; Ma Miling; Li Youquan; Xu Zongke; Fu Yuguang; Yang Jifei; Guan Guiquan; Niu Qingli; Luo Jianxun; Yin Hong

    2011-01-01

    Abstract Background Lyme disease caused by Borrelia burgdorferi sensu lato complex is an important endemic zoonosis whose distribution is closely related to the main ixodid tick vectors. In China, isolated cases of Lyme disease infection of humans have been reported in 29 provinces. Ticks, especially ixodid ticks are abundant and a wide arrange of Borrelia natural reservoirs are present. In this study, we developed a reverse line blot (RLB) to identify Borrelia spp. in ticks collected from sh...

  9. Two ways of experimental infection of Ixodes ricinus ticks (Acari: Ixodidae) with spirochetes of Borrelia burgdorferi sensu lato complex

    Czech Academy of Sciences Publication Activity Database

    Fišerová, Lenka; Černá, Kateřina; Horká, Helena; Kopecký, Jan

    2008-01-01

    Roč. 55, č. 2 (2008), s. 150-154. ISSN 0015-5683 R&D Projects: GA ČR GA524/05/0811; GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z60220518 Keywords : Lyme disease spirochete * Borrelia burgdorferi * Borrelia baronii * Borrelia afzelii * tick * Ixodes ricinus Subject RIV: EC - Immunology Impact factor: 1.307, year: 2008

  10. Lyme Disease: antibodies against Borrelia burgdorferi in farm workers in Argentina Doença de Lyme: anticorpos anti Borrelia burgdorferi nos trabalhadores rurais da Argentina

    Directory of Open Access Journals (Sweden)

    Nestor Oscar Stanchi

    1993-08-01

    Full Text Available Lyme Disease is a tick-borne (specially by Ixodes ticks immune-mediated inflammatory disorder caused by a newly recognize spirochete, Borrelia burgdorferi. Indirect fluorescent antibody (IF staining methods and enzyme-linked immunosorbent assay are frequently relied upon to confirm Lyme borreliosis infections. Although serologic testing for antibodies has limitations, it is still the only practical means of confirming B. burgdorferi infections. Because we have no previous report of Lyme disease in human inhabitants in Argentina, a study was designed as a seroepidemiologic investigation of the immune response to B. burgdorferi in farm workers of Argentina with arthritis symptoms. Three out of 28 sera were positive (#1,5 and 9. Serum # 1 was positive for Immunoglobulin G at dilution 1:320, serum # 5 and # 9 both to dilution 1:160; while for Immunoglobulin M all (#1, 5 and 9 were positive at low dilution (1:40 using IF. The results showed that antibodies against B. burgdorferi are present in an Argentinian population. Thus caution should be exercised in the clinical interpretation of arthritis until the presence of B. burgdorferi be confirmed by culture in specific media.A doença de Lyme, é uma desordem inflamatória, intermediada pelo sistema imunogênico, transmitida por carrapatos (especialmente do Gênero Ixodes e causada por uma espiroqueta recentemente descoberta, a Borrelia burgdorferi. A técnica de Imunofluorescência Indireta (IF é com freqüência usada para confirmar o diagnóstico da infecção por este microrganismo. Embora os métodos práticos tenham limitações, é no entanto o único método prático para seu diagnóstico. Devido a não existência de registros prévios dessa doença na Argentina, foi realizada pesquisa seroepidemiológica para determinar a presença de imunoglobulinas nos trabalhadores rurais da Argentina, com sintomas de artrite. Sobre um total de 28 soros analisados, 3 resultaram positivos (o soro número 1

  11. Further delineation of Borrelia burgdorferi Restriction-Modification system and understanding antibiotic resistance in Borrelia afzelii

    OpenAIRE

    DEMCHYSHYN, Stepan Stepanovych

    2015-01-01

    The aim of this thesis was to further the understanding of restriction-modification and the homology within the restriction-modification genes in various Borrelia species including Borrelia afzelii as well as to investigate spontaneous antibiotic resistance within this particular Borrelia species.

  12. NMR structure of an acyl-carrier protein from Borrelia burgdorferi

    OpenAIRE

    Barnwal, Ravi P.; Van Voorhis, Wesley C.; Varani, G

    2011-01-01

    Nearly complete resonance assignment and the high-resolution NMR structure of the acyl-carrier protein from Borrelia burgdorferi, a target of the Seattle Structural Genomics Center for Infectious Disease (SSGCID) structure-determination pipeline, are reported. This protein was chosen as a potential target for drug-discovery efforts because of its involvement in fatty-acid biosynthesis, an essential metabolic pathway, in bacteria. It was possible to assign >98% of backbone resonances and >92% ...

  13. Cryoelectron tomography reveals the sequential assembly of bacterial flagella in Borrelia burgdorferi

    OpenAIRE

    Zhao, Xiaowei; Zhang, Kai; Boquoi, Tristan; Hu, Bo; Motaleb, M. A.; Miller, Kelly A.; James, Milinda E; Charon, Nyles W.; Manson, Michael D.; Norris, Steven J; Li, Chunhao; Liu, Jun

    2013-01-01

    Periplasmic flagella are essential for the distinctive morphology, motility, and infectious life cycle of the Lyme disease spirochete Borrelia burgdorferi. In this study, we genetically trapped intermediates in flagellar assembly and determined the 3D structures of the intermediates to 4-nm resolution by cryoelectron tomography. We provide structural evidence that secretion of rod substrates triggers remodeling of the central channel in the flagellar secretion apparatus from a closed to an op...

  14. Transcriptional Regulation of the Borrelia burgdorferi Antigenically Variable VlsE Surface Protein

    OpenAIRE

    Bykowski, Tomasz; Babb, Kelly; von Lackum, Kate; Riley, Sean P.; Norris, Steven J; Stevenson, Brian

    2006-01-01

    The Lyme disease agent Borrelia burgdorferi can persistently infect humans and other animals despite host active immune responses. This is facilitated, in part, by the vls locus, a complex system consisting of the vlsE expression site and an adjacent set of 11 to 15 silent vls cassettes. Segments of nonexpressed cassettes recombine with the vlsE region during infection of mammalian hosts, resulting in combinatorial antigenic variation of the VlsE outer surface protein. We now demonstrate that...

  15. Serologic survey of the wild boar (Sus scrofa) for Borrelia burgdorferi sensu lato

    Czech Academy of Sciences Publication Activity Database

    Juřicová, Zina; Hubálek, Zdeněk

    2009-01-01

    Roč. 9, č. 5 (2009), s. 479-482. ISSN 1530-3667 R&D Projects: GA ČR GA524/96/1059 Grant ostatní: 6th Framework Programme EC(XE) GOCE-2003-010284 EDEN Institutional research plan: CEZ:AV0Z60930519 Keywords : Borrelia burgdorferi * Sus scrofa * serosurvey * Czech Republic Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 2.607, year: 2009

  16. SERUM ANTIBODIES TO WHOLE-CELL AND RECOMBINANT ANTIGENS OF BORRELIA BURGDORFERI IN COTTONTAIL RABBITS

    OpenAIRE

    Magnarelli, Louis A.; Norris, Steven J; Fikrig, Erol

    2012-01-01

    Archived serum samples, from 95 eastern cottontail rabbits (Sylvilagus floridanus) captured in New York, New York, USA and Millbrook, New York, USA, during 1985–86, were analyzed in solid-phase enzyme-linked immunosorbent assays (ELISA) for total and class-specific immunoglobulin (Ig) M antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Using a polyvalent conjugate, rabbit sera contained antibodies to whole-cell and recombinant antigens (protein [p]35, p37...

  17. First Isolation and Cultivation of Borrelia burgdorferi Sensu Lato from Missouri

    OpenAIRE

    Oliver, J H; Kollars, T. M.; Chandler, F W; James, A. M.; Masters, E. J.; Lane, R S; Huey, L. O.

    1998-01-01

    Five Borrelia burgdorferi sensu lato isolates from Missouri are described. This represents the first report and characterization of such isolates from that state. The isolates were obtained from either Ixodes dentatus or Amblyomma americanum ticks that had been feeding on cottontail rabbits (Sylvilagus floridanus) from a farm in Bollinger County, Mo., where a human case of Lyme disease had been reported. All isolates were screened immunologically by indirect immunofluorescence by using monocl...

  18. Molecular Typing of Borrelia burgdorferi Sensu Lato: Taxonomic, Epidemiological, and Clinical Implications

    OpenAIRE

    Wang, Guiqing; Dam, Alje P. van; Schwartz, Ira; Dankert, Jacob

    1999-01-01

    Borrelia burgdorferi sensu lato, the spirochete that causes human Lyme borreliosis (LB), is a genetically and phenotypically divergent species. In the past several years, various molecular approaches have been developed and used to determine the phenotypic and genetic heterogeneity within the LB-related spirochetes and their potential association with distinct clinical syndromes. These methods include serotyping, multilocus enzyme electrophoresis, DNA-DNA reassociation analysis, rRNA gene res...

  19. Genomic fingerprinting of Borrelia burgdorferi sensu lato by pulsed-field gel electrophoresis.

    OpenAIRE

    1993-01-01

    A total of 46 Borrelia burgdorferi sensu lato isolates that were isolated from patients with Lyme borreliosis and infected animals or were extracted from ticks of the genus Ixodes were analyzed. Large restriction fragment patterns obtained after cleavage of genomic DNAs with MluI were analyzed by pulsed-field gel electrophoresis (PFGE). To eliminate the contribution of plasmid DNA, only fragments greater than 70 kb were used for the analysis. The results indicated that each of the 14 B. burgd...

  20. Anaphylactoid reaction caused by sodium ceftriaxone in two horses experimentally infected by Borrelia burgdorferi

    OpenAIRE

    Basile, Roberta Carvalho; Rivera, Gabriela Gomes; Del Rio, Lara Antoniassi; de Bonis, Talissa Camargo Mantovani; do Amaral, Gabriel Paiva Domingues; Giangrecco, Edson; Ferraz, Guilherme; Yoshinari, Natalino Hajime; Canola, Paulo Aléscio; Queiroz Neto, Antonio

    2015-01-01

    Background Lyme borreliosis is a disease transmitted by ticks to mammals, especially in horses and humans. Caused by a spirochete Borrelia burgdorferi, it can result in lameness, arthritis, carditis, dermatitis and neurological signs. Anaphylactoid reactions are severe responses caused by direct action of substances (drugs, toxins), which can pose risks to life. Still poorly documented in horses, these reactions are caused by the effects of inflammatory mediators such as histamine, kinins and...

  1. Probable late lyme disease: a variant manifestation of untreated Borrelia burgdorferi infection

    OpenAIRE

    Aucott John N; Seifter Ari; Rebman Alison W

    2012-01-01

    Abstract Background Lyme disease, a bacterial infection with the tick-borne spirochete Borrelia burgdorferi, can cause early and late manifestations. The category of probable Lyme disease was recently added to the CDC surveillance case definition to describe patients with serologic evidence of exposure and physician-diagnosed disease in the absence of objective signs. We present a retrospective case series of 13 untreated patients with persistent symptoms of greater than 12 weeks duration who...

  2. Uncoordinated phylogeography of Borrelia burgdorferi and its tick vector, Ixodes scapularis

    OpenAIRE

    Humphrey, Parris T; Caporale, Diane A.; Brisson, Dustin

    2010-01-01

    Vector-borne microbes necessarily co-occur with their hosts and vectors, but the degree to which they share common evolutionary or biogeographic histories remains unexplored. We examine the congruity of the evolutionary and biogeographic histories of the Lyme disease system, the most prevalent vector-borne disease in North America. In the Eastern and Midwestern US, Ixodes scapularis ticks are the primary vectors of Borrelia burgdorferi, the bacterium that causes Lyme disease. Our phylogeograp...

  3. Biochemical and immunological characterization of the surface proteins of Borrelia burgdorferi.

    OpenAIRE

    Luft, B J; Jiang, W.; Munoz, P.; Dattwyler, R J; Gorevic, P D

    1989-01-01

    The immunodominant proteins and glycoproteins of Borrelia burgdorferi were analyzed by one-dimensional (1D) and 2D gel electrophoresis. More than 100 polypeptide species could be detected on silver-stained 2D gels. Separation of sonic extracts of the organism by differential centrifugation (100,000 X g) revealed several of the major proteins to reside predominantly within the pellet fraction. The antigenicity of the individual polypeptides was determined by Western (immuno-) blot analysis wit...

  4. Mapping the major antigenic domains of the native flagellar antigen of Borrelia burgdorferi.

    OpenAIRE

    Jiang, W.; Luft, B J; Schubach, W; Dattwyler, R J; Gorevic, P D

    1992-01-01

    Purified flagellar protein (p41) of Borrelia burgdorferi (strain B31) was subjected to chemical cleavage with hydroxylamine or proteolysis with V8 protease, endoproteinase Asp-N, or alpha-chymotrypsin. The resulting polypeptides were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their positions in the published DNA sequence of the p41 protein were determined by amino-terminal sequencing and amino acid analysis. Epitope specificities of antibody binding by a mono...

  5. Outer surface protein B is critical for Borrelia burgdorferi adherence and survival within Ixodes ticks.

    Directory of Open Access Journals (Sweden)

    Girish Neelakanta

    2007-03-01

    Full Text Available Survival of Borrelia burgdorferi in ticks and mammals is facilitated, at least in part, by the selective expression of lipoproteins. Outer surface protein (Osp A participates in spirochete adherence to the tick gut. As ospB is expressed on a bicistronic operon with ospA, we have now investigated the role of OspB by generating an OspB-deficient B. burgdorferi and examining its phenotype throughout the spirochete life cycle. Similar to wild-type isolates, the OspB-deficient B. burgdorferi were able to readily infect and persist in mice. OspB-deficient B. burgdorferi were capable of migrating to the feeding ticks but had an impaired ability to adhere to the tick gut and survive within the vector. Furthermore, the OspB-deficient B. burgdorferi bound poorly to tick gut extracts. The complementation of the OspB-deficient spirochete in trans, with a wild-type copy of ospB gene, restored its ability to bind tick gut. Taken together, these data suggest that OspB has an important role within Ixodes scapularis and that B. burgdorferi relies upon multiple genes to efficiently persist in ticks.

  6. Prevalence of Borrelia burgdorferi sensu lato in rodents from Gansu, northwestern China

    Directory of Open Access Journals (Sweden)

    Liu Zengjia

    2010-05-01

    Full Text Available Abstract Background Lyme disease is a multi-organ infection disease caused by Borrelia burgdorferi sensu lato. Lyme disease was first documented in north-east China in 1986. Since then more than 20 provinces in China were confirmed the existence of nature foci of Lyme disease. In the present study, a molecular epidemiological survey was conducted to investigate the presence of Borrelia burgdorferi sensu lato in rodents from Gansu Province for the first time. Result A total of 140 rodents of 7 species were examined for Borrelia burgdorferi sensu lato. by nested-PCR and culture isolation. The overall infection rate was 22.86%. Two rodent species most frequently trapped were responsible for all positive. 3 strains were isolated from Apodemus agrarius, which belonged to B. garinii, 1 strain isolated from Rattus losea was identified as B. afzelii. Conclusion The study firstly showed the role of rodents in maintaining the pathogen of Lyme disease in the environment from Gansu Province and there existed at least two genotypes of Lyme disease spirochaetes in rodents.

  7. Differences in Genotype, Clinical Features, and Inflammatory Potential of Borrelia burgdorferi sensu stricto Strains from Europe and the United States.

    Science.gov (United States)

    Cerar, Tjasa; Strle, Franc; Stupica, Dasa; Ruzic-Sabljic, Eva; McHugh, Gail; Steere, Allen C; Strle, Klemen

    2016-05-01

    Borrelia burgdorferi sensu stricto isolates from patients with erythema migrans in Europe and the United States were compared by genotype, clinical features of infection, and inflammatory potential. Analysis of outer surface protein C and multilocus sequence typing showed that strains from these 2 regions represent distinct genotypes. Clinical features of infection with B. burgdorferi in Slovenia were similar to infection with B. afzelii or B. garinii, the other 2 Borrelia spp. that cause disease in Europe, whereas B. burgdorferi strains from the United States were associated with more severe disease. Moreover, B. burgdorferi strains from the United States induced peripheral blood mononuclear cells to secrete higher levels of cytokines and chemokines associated with innate and Th1-adaptive immune responses, whereas strains from Europe induced greater Th17-associated responses. Thus, strains of the same B. burgdorferi species from Europe and the United States represent distinct clonal lineages that vary in virulence and inflammatory potential. PMID:27088349

  8. Vectors as Epidemiological Sentinels: Patterns of Within-Tick Borrelia burgdorferi Diversity.

    Science.gov (United States)

    Walter, Katharine S; Carpi, Giovanna; Evans, Benjamin R; Caccone, Adalgisa; Diuk-Wasser, Maria A

    2016-07-01

    Hosts including humans, other vertebrates, and arthropods, are frequently infected with heterogeneous populations of pathogens. Within-host pathogen diversity has major implications for human health, epidemiology, and pathogen evolution. However, pathogen diversity within-hosts is difficult to characterize and little is known about the levels and sources of within-host diversity maintained in natural populations of disease vectors. Here, we examine genomic variation of the Lyme disease bacteria, Borrelia burgdorferi (Bb), in 98 individual field-collected tick vectors as a model for study of within-host processes. Deep population sequencing reveals extensive and previously undocumented levels of Bb variation: the majority (~70%) of ticks harbor mixed strain infections, which we define as levels Bb diversity pre-existing in a diverse inoculum. Within-tick diversity is thus a sample of the variation present within vertebrate hosts. Within individual ticks, we detect signatures of positive selection. Genes most commonly under positive selection across ticks include those involved in dissemination in vertebrate hosts and evasion of the vertebrate immune complement. By focusing on tick-borne Bb, we show that vectors can serve as epidemiological and evolutionary sentinels: within-vector pathogen diversity can be a useful and unbiased way to survey circulating pathogen diversity and identify evolutionary processes occurring in natural transmission cycles. PMID:27414806

  9. Decreased Electroporation Efficiency in Borrelia burgdorferi Containing Linear Plasmids lp25 and lp56: Impact on Transformation of Infectious B. burgdorferi

    OpenAIRE

    Matthew B Lawrenz; Kawabata, Hiroki; Purser, Joye E.; Norris, Steven J

    2002-01-01

    The presence of the linear plasmids lp25 and lp56 of Borrelia burgdorferi B31 was found to dramatically decrease the rate of transformation by electroporation with the shuttle vector pBSV2, an autonomously replicating plasmid that confers kanamycin resistance (P. E. Stewart, R. Thalken, J. L. Bono, and P. Rosa, Mol. Microbiol. 39:714-721, 2001). B. burgdorferi B31 clones had transformation efficiencies that were either low, intermediate, or high, and this phenotype correlated with the presenc...

  10. ELISA-Based Measurement of Antibody Responses and PCR-Based Detection Profiles Can Distinguish between Active Infection and Early Clearance of Borrelia burgdorferi

    OpenAIRE

    Lazarus, John J.; McCarter, Akisha L.; Kari Neifer-Sadhwani; R. Mark Wooten

    2012-01-01

    Borrelia burgdorferi is a spirochetal bacterium that causes Lyme disease. These studies address whether current research methods using either ELISA to detect seroconversion to B. burgdorferi antigens or PCR quantification of bacterial DNA within tissues can accurately distinguish between a productive infection versus a B. burgdorferi exposure that is rapidly cleared by the innate responses. Mice receiving even minimal doses of live B. burgdorferi produced significantly more B. burgdorferi-spe...

  11. Bird-feeding ticks transstadially transmit Borrelia burgdorferi that infect Syrian hamsters.

    Science.gov (United States)

    Anderson, J F; Magnarelli, L A; Stafford, K C

    1990-01-01

    Bird-feeding Ixodes dammini ticks were documented for the first time to successfully molt and transstadially pass Borrelia burgdorferi spirochetes that were indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from the type B31 strain. Forty-six of 73 blood-engorged larvae and 50 of 66 fully-fed nymphs, removed from wild-caught birds, successfully molted. Borreliae were isolated from 21 of 78 partially- and fully-fed larvae off birds, including six specimens that molted. Spirochete-positive cultures also were obtained from 35 of 60 partially- and fully-fed nymphs that had fed from birds, including 20 nymphs that molted into adult ticks. Transstadially passed borreliae by bird-feeding larval and nymphal I. dammini were infectious to hamsters, leading us to suggest that these ticks are capable of subsequently transmitting infectious spirochetes to mammals, including humans. An isolated of B. burgdorferi, recovered from a bird-feeding larval Ixodes dentatus, was indistinguishable by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from the B31 strain. This isolate, unlike another from I. dentatus off a cottontail rabbit (Sylvilagus floridanus), had a protein band with a molecular weight of approximately 31,000 that reacted with murine monoclonal antibodies H3TS and H5332 in western blot analysis. Thus, closely related borreliae are present in both I. dentatus and I. dammini. PMID:2304189

  12. Proteomic Analysis of Lyme Disease: Global Protein Comparison of Three Strains of Borrelia burgdorferi

    Energy Technology Data Exchange (ETDEWEB)

    Jacobs, Jon M.; Yang, Xiaohua; Luft, Benjamin J.; Dunn, John J.; Camp, David G.; Smith, Richard D.

    2005-04-01

    The Borrelia burgdorferi spirochete is the causative agent of Lyme disease, the most common tick-borne disease in the United States. It has been studied extensively to help understand its pathogenicity of infection and how it can persist in different mammalian hosts. We report the proteomic analysis of the archetype B. burgdorferi B31 strain and two other strains (ND40, and JD-1) having different Borrelia pathotypes using strong cation exchange fractionation of proteolytic peptides followed by high-resolution, reversed phase capillary liquid chromatography coupled with ion trap tandem mass spectrometric (LC-MS/MS) analysis. Protein identification was facilitated by the availability of the complete B31 genome sequence. A total of 665 Borrelia proteins were identified representing ~38 % coverage of the theoretical B31 proteome. A significant overlap was observed between the identified proteins in direct comparisons between any two strains (>72%), but distinct differences were observed among identified hypothetical and outer membrane proteins of the three strains. Such a concurrent proteomic overview of three Borrelia strains based upon only the B31 genome sequence is shown to provide significant insights into the presence or absence of specific proteins and a broad overall comparison among strains.

  13. Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected from migratory birds in Southern Norway

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    Skarpaas Tone

    2010-11-01

    Full Text Available Abstract Background Borrelia burgdorferi sensu lato (s.l. are the causative agent for Lyme borreliosis (LB, the most common tick-borne disease in the northern hemisphere. Birds are considered important in the global dispersal of ticks and tick-borne pathogens through their migration. The present study is the first description of B. burgdorferi prevalence and genotypes in Ixodes ricinus ticks feeding on birds during spring and autumn migration in Norway. Methods 6538 migratory birds were captured and examined for ticks at Lista Bird Observatory during the spring and the autumn migration in 2008. 822 immature I. ricinus ticks were collected from 215 infested birds. Ticks were investigated for infection with B. burgdorferi s.l. by real-time PCR amplification of the 16S rRNA gene, and B. burgdorferi s.l. were thereafter genotyped by melting curve analysis after real-time PCR amplification of the hbb gene, or by direct sequencing of the PCR amplicon generated from the rrs (16S-rrl (23S intergenetic spacer. Results B. burgdorferi s.l. were detected in 4.4% of the ticks. The most prevalent B. burgdorferi genospecies identified were B. garinii (77.8%, followed by B.valaisiana (11.1%, B. afzelii (8.3% and B. burgdorferi sensu stricto (2.8%. Conclusion Infection rate in ticks and genospecies composition were similar in spring and autumn migration, however, the prevalence of ticks on birds was higher during spring migration. The study supports the notion that birds are important in the dispersal of ticks, and that they may be partly responsible for the heterogeneous distribution of B. burgdorferi s.l. in Europe.

  14. Borrelia carolinensis sp. nov., a new (14th) member of the Borrelia burgdorferi Sensu Lato complex from the southeastern region of the United States

    Czech Academy of Sciences Publication Activity Database

    Rudenko, Natalia; Golovchenko, Maryna; Grubhoffer, Libor; Oliver, J. H., Jr.

    2009-01-01

    Roč. 47, č. 1 (2009), s. 134-141. ISSN 0095-1137 R&D Projects: GA MŠk(CZ) LC06009; GA ČR(CZ) GA524/06/1479 Institutional research plan: CEZ:AV0Z60220518 Keywords : multilocus sequence analysis * B. burgdorferi sl complex * new borrelia species * Borrelia carolinensis Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 4.162, year: 2009

  15. Imbalanced presence of Borrelia burgdorferi s.l. multilocus sequence types in clinical manifestations of Lyme borreliosis.

    Science.gov (United States)

    Coipan, E Claudia; Jahfari, Setareh; Fonville, Manoj; Oei, G Anneke; Spanjaard, Lodewijk; Takumi, Katsuhisa; Hovius, Joppe W R; Sprong, Hein

    2016-08-01

    In this study we used typing based on the eight multilocus sequence typing scheme housekeeping genes (MLST) and 5S-23S rDNA intergenic spacer (IGS) to explore the population structure of Borrelia burgdorferi sensu lato isolates from patients with Lyme borreliosis (LB) and to test the association between the B. burgdorferi s.l. sequence types (ST) and the clinical manifestations they cause in humans. Isolates of B. burgdorferi from 183 LB cases across Europe, with distinct clinical manifestations, and 257 Ixodes ricinus lysates from The Netherlands, were analyzed for this study alone. For completeness, we incorporated in our analysis also 335 European B. burgdorferi s.l. MLST profiles retrieved from literature. Borrelia afzelii and Borrelia bavariensis were associated with human cases of LB while Borrelia garinii, Borrelia lusitaniae and Borrelia valaisiana were associated with questing I. ricinus ticks. B. afzelii was associated with acrodermatitis chronica atrophicans, while B. garinii and B. bavariensis were associated with neuroborreliosis. The samples in our study belonged to 251 different STs, of which 94 are newly described, adding to the overall picture of the genetic diversity of Borrelia genospecies. The fraction of STs that were isolated from human samples was significantly higher for the genospecies that are known to be maintained in enzootic cycles by mammals (B. afzelii, B. bavariensis, and Borrelia spielmanii) than for genospecies that are maintained by birds (B. garinii and B. valaisiana) or lizards (B. lusitaniae). We found six multilocus sequence types that were significantly associated to clinical manifestations in humans and five IGS haplotypes that were associated with the human LB cases. While IGS could perform just as well as the housekeeping genes in the MLST scheme for predicting the infectivity of B. burgdorferi s.l., the advantage of MLST is that it can also capture the differential invasiveness of the various STs. PMID:27125686

  16. Borrelia burgdorferi in ticks and dogs in the province of Vojvodina, Serbia*

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    Savić S.

    2010-12-01

    Full Text Available Lyme disease is a tick borne zoonotic infection, caused by Borrelia burgdorferi s.l. bacteria. For the transmission of the disease, the presence of ticks is a prerequisite. Lyme borreliosis mostly occurs in people and dogs, but it may occur in other animals. Ticks which carry B. burgdorferi s.l. in Serbia are of the Ixodes ricinus specis. In Serbia, Lyme disease was detected for the first time in the late ‘80-es. In dogs, clinical symptoms may occur even months after a tick bite, and include weakness, lymphadenopathy, fever, lameness, arthritis, etc. In our survey, we have observed tick and dog populations in the province of Vojvodina (northern part of Serbia. I. ricinus ticks were collected and examined for the presence of B. burgdorferi s.l. in several chosen locations. In addition, blood samples were collected from house dogs and pets from the same locations, and analyzed for the presence of antibodies specific for B. burgdorferi s.l. The results showed a mean infection of ticks of 22.12 %, and a mean seroprevalence of Lyme disease in dogs of 25.81 %. We conclude that in Vojvodina there is an actual risk of Lyme borreliosis for other animals and humans, because of the persistence of B. burgdorferi s.l. in both tick and dog populations.

  17. Lyme Disease: antibodies against Borrelia burgdorferi in farm workers in Argentina

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    Stanchi Nestor Oscar

    1993-01-01

    Full Text Available Lyme Disease is a tick-borne (specially by Ixodes ticks immune-mediated inflammatory disorder caused by a newly recognize spirochete, Borrelia burgdorferi. Indirect fluorescent antibody (IF staining methods and enzyme-linked immunosorbent assay are frequently relied upon to confirm Lyme borreliosis infections. Although serologic testing for antibodies has limitations, it is still the only practical means of confirming B. burgdorferi infections. Because we have no previous report of Lyme disease in human inhabitants in Argentina, a study was designed as a seroepidemiologic investigation of the immune response to B. burgdorferi in farm workers of Argentina with arthritis symptoms. Three out of 28 sera were positive (#1,5 and 9. Serum # 1 was positive for Immunoglobulin G at dilution 1:320, serum # 5 and # 9 both to dilution 1:160; while for Immunoglobulin M all (#1, 5 and 9 were positive at low dilution (1:40 using IF. The results showed that antibodies against B. burgdorferi are present in an Argentinian population. Thus caution should be exercised in the clinical interpretation of arthritis until the presence of B. burgdorferi be confirmed by culture in specific media.

  18. Lyme Disease: antibodies against Borrelia burgdorferi in farm workers in Argentina

    Directory of Open Access Journals (Sweden)

    Nestor Oscar Stanchi

    1993-08-01

    Full Text Available Lyme Disease is a tick-borne (specially by Ixodes ticks immune-mediated inflammatory disorder caused by a newly recognize spirochete, Borrelia burgdorferi. Indirect fluorescent antibody (IF staining methods and enzyme-linked immunosorbent assay are frequently relied upon to confirm Lyme borreliosis infections. Although serologic testing for antibodies has limitations, it is still the only practical means of confirming B. burgdorferi infections. Because we have no previous report of Lyme disease in human inhabitants in Argentina, a study was designed as a seroepidemiologic investigation of the immune response to B. burgdorferi in farm workers of Argentina with arthritis symptoms. Three out of 28 sera were positive (#1,5 and 9. Serum # 1 was positive for Immunoglobulin G at dilution 1:320, serum # 5 and # 9 both to dilution 1:160; while for Immunoglobulin M all (#1, 5 and 9 were positive at low dilution (1:40 using IF. The results showed that antibodies against B. burgdorferi are present in an Argentinian population. Thus caution should be exercised in the clinical interpretation of arthritis until the presence of B. burgdorferi be confirmed by culture in specific media.

  19. Cinética do crescimento de Borrelia burgdorferi (Spirochaetaceae em diferentes meios de cultivo Cinetic growth of Borrelia burgdorferi (Spirochaetacease in different culture media

    Directory of Open Access Journals (Sweden)

    Angela de Oliveira

    2004-06-01

    Full Text Available Estudou-se a cinética de crescimento de Borrelia burgdorferi, por um período de 3 meses, utilizando os seguintes oito meios de cultivo : (1 BSK adicionado de soro de coelho, (2 BSK adicionado de soro de suíno, (3 BSK adicionado de soro de suíno + 5 fluorouracil, (4 PMR, (5 CTB, (6 Dubos, (7 Caldo Brucella e (8 BHI. Todos os meios foram preparados assepticamente e mantidos em tubos de ensaio com capacidade para 10 ml. Para cada meio, o inoculo foi padronizado para conter no início 10² espiroquetas para cada 0,1 ml de cultivo. O monitoramento do crescimento foi feito contando-se o total de espiroquetas em 0,1 ml do meio entre lâmina de microscopia e lamínula com dimen sões de 10x30mm, tendo sido utilizado microscópio de campo escuro. A contagem foi realizada durante 14 semanas, tendo sido diária nos primeiros 12 dias e semanal a partir desta data. Houve crescimento de B. burgdorferi em todos meios testados, com melhor performance para três deles: BSK adicionado de soro de coelho, BSK adicionado de soro de suíno + 5 fluorouracil e meio CTB. Observou-se crescimento de B. burgdorferi a partir da 4ª semana, atingindo o platô de crescimento entre a 8ª e 12ª semanas, quando começou a exaustão do meio de cultivo. Formas císticas de B. burgdorferi foram observadas em todos os meios testados.The cinetic of growth of Borrelia burgdorferi was studied during a 3-month period, using the following 8 culture media: (1 rabbit serum BSK, (2 swine serum BSK, (3 swine serum BSK+5 fluorouracil, (4 PMR, (5 CTB, (6 Dubos, (7 Brucella broth and (8 BHI. All media were prepared aseptically and were maintained in culture tubes of 10 ml capacity. For each medium, the inoculum was standardized to contain initially 10² spirochetes for each 0.1 ml of culture. The growth was monitorized by counting the total number of spirochetes in 0.1ml of medium in a dark field microscope, using a 10x30 mm cover slip. For the first 12 days, counting was done each 24

  20. Resurgence of persisting non-cultivable Borrelia burgdorferi following antibiotic treatment in mice.

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    Emir Hodzic

    Full Text Available The agent of Lyme borreliosis, Borrelia burgdorferi, evades host immunity and establishes persistent infections in its varied mammalian hosts. This persistent biology may pose challenges to effective antibiotic treatment. Experimental studies in dogs, mice, and non-human primates have found persistence of B. burgdorferi DNA following treatment with a variety of antibiotics, but persisting spirochetes are non-cultivable. Persistence of B. burgdorferi DNA has been documented in humans following treatment, but the significance remains unknown. The present study utilized a ceftriaxone treatment regimen in the C3H mouse model that resulted in persistence of non-cultivable B. burgdorferi in order to determine their long-term fate, and to examine their effects on the host. Results confirmed previous studies, in which B. burgdorferi could not be cultured from tissues, but low copy numbers of B. burgdorferi flaB DNA were detectable in tissues at 2, 4 and 8 months after completion of treatment, and the rate of PCR-positive tissues appeared to progressively decline over time. However, there was resurgence of spirochete flaB DNA in multiple tissues at 12 months, with flaB DNA copy levels nearly equivalent to those found in saline-treated mice. Despite the continued non-cultivable state, RNA transcription of multiple B. burgdorferi genes was detected in host tissues, flaB DNA was acquired by xenodiagnostic ticks, and spirochetal forms could be visualized within ticks and mouse tissues by immunofluorescence and immunohistochemistry, respectively. A number of host cytokines were up- or down-regulated in tissues of both saline- and antibiotic-treated mice in the absence of histopathology, indicating host response to the presence of non-cultivable, despite the lack of inflammation in tissues.

  1. Role of Acetyl-Phosphate in Activation of the Rrp2-RpoN-RpoS Pathway in Borrelia burgdorferi

    Science.gov (United States)

    Xu, Haijun; Caimano, Melissa J.; Lin, Tao; He, Ming; Radolf, Justin D.; Norris, Steven J.; Gheradini, Frank; Wolfe, Alan J.; Yang, X. Frank

    2010-01-01

    Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ54–σS sigma factor cascade), plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl∼P), the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase) pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl∼P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl∼P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl∼P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl∼P can serve as a global signal in bacterial pathogenesis. PMID:20862323

  2. Role of acetyl-phosphate in activation of the Rrp2-RpoN-RpoS pathway in Borrelia burgdorferi.

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    Haijun Xu

    Full Text Available Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ(54-σ(S sigma factor cascade, plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s by which the upstream activator and response regulator Rrp2 is activated remains unclear. Here, we show that none of the histidine kinases present in the B. burgdorferi genome are required for the activation of Rrp2. Instead, we present biochemical and genetic evidence that supports the hypothesis that activation of the Rrp2-RpoN-RpoS pathway occurs via the small, high-energy, phosphoryl-donor acetyl phosphate (acetyl∼P, the intermediate of the Ack-Pta (acetate kinase-phosphate acetyltransferase pathway that converts acetate to acetyl-CoA. Supplementation of the growth medium with acetate induced activation of the Rrp2-RpoN-RpoS pathway in a dose-dependent manner. Conversely, the overexpression of Pta virtually abolished acetate-induced activation of this pathway, suggesting that acetate works through acetyl∼P. Overexpression of Pta also greatly inhibited temperature and cell density-induced activation of RpoS and OspC, suggesting that these environmental cues affect the Rrp2-RpoN-RpoS pathway by influencing acetyl∼P. Finally, overexpression of Pta partially reduced infectivity of B. burgdorferi in mice. Taken together, these findings suggest that acetyl∼P is one of the key activating molecule for the activation of the Rrp2-RpoN-RpoS pathway and support the emerging concept that acetyl∼P can serve as a global signal in bacterial pathogenesis.

  3. Seroprevalence of Leptospira spp. and Borrelia burgdorferi sensu lato in Italian horses.

    Science.gov (United States)

    Ebani, Valentina V; Bertelloni, Fabrizio; Pinzauti, Paolo; Cerri, Domenico

    2012-01-01

    The aim of the study was to determine the seroprevalence of Leptospira spp. and Borrelia burgdorferi sensu lato in healthy horses living in 7 provinces of central Italy. In the period 2007-2009, sera from 386 horses were tested by microagglutination test (MAT) to detect antibodies to Leptospira spp., employing the following serovars as antigens: Bratislava, Ballum, Canicola, Icterohaemorrhagiae, Grippotyphosa, Hardjo, Pomona, Tarassovi. 3 animals were positive for the serovars Icterohaemorrhagiae, 2 to Bratislava, and 1 to Pomona, for a total 1.5% seroprevalence. All sera were examined by immunofluorence antibody test (IFAT) to reveal anti-B. burgdorferi s.l. antibodies. 94 (24.3%) horses were positive with antibody titres ranging from 1:64 to 1:1,024. The seroprevalence was significantly higher in >10 year-old horses compared to younger subjects. No significant differences in the mean seroprevalence were observed in the respective years. The total mean seroprevalence were strictly related to the environmental conditions of the areas in which the horses lived. No cross-reactions between Leptospira and Borrelia were observed. This is the first serological survey on antibodies to B. burgdorferi s.l. in Italian horses. PMID:22742794

  4. Positive IgG Western Blot for Borrelia burgdorferi in Colombia

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    Palacios Ricardo

    1999-01-01

    Full Text Available In order to evaluate the presence of specific IgG antibodies to Borrelia burgdorferi in patients with clinical manifestations associated with Lyme borreliosis in Cali, Colombia, 20 serum samples from patients with dermatologic signs, one cerebrospinal fluid (CSF sample from a patient with chronic neurologic and arthritic manifestations, and twelve serum samples from individuals without clinical signs associated with Lyme borreliosis were analyzed by IgG Western blot. The results were interpreted following the recommendations of the Centers for Diseases Control and Prevention (CDC for IgG Western blots. Four samples fulfilled the CDC criteria: two serum specimens from patients with morphea (localized scleroderma, the CSF from the patient with neurologic and arthritic manifestations, and one of the controls. Interpretation of positive serology for Lyme disease in non-endemic countries must be cautious. However these results suggest that the putative "Lyme-like" disease may correlate with positivity on Western blots, thus raising the possibility that a spirochete genospecies distinct from B. burgdorferi sensu stricto, or a Borrelia species other than B. burgdorferi sensu lato is the causative agent. Future work will focus on a survey of the local tick and rodent population for evidence of spirochete species that could be incriminated as the etiologic agent.

  5. Disruption of bbe02 by Insertion of a Luciferase Gene Increases Transformation Efficiency of Borrelia burgdorferi and Allows Live Imaging in Lyme Disease Susceptible C3H Mice.

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    Kamfai Chan

    Full Text Available Lyme disease is the most prevalent tick-borne disease in North America and Europe. The causative agent, Borrelia burgdorferi persists in the white-footed mouse. Infection with B. burgdorferi can cause acute to persistent multisystemic Lyme disease in humans. Some disease manifestations are also exhibited in the mouse model of Lyme disease. Genetic manipulation of B. burgdorferi remains difficult. First, B. burgdorferi contains a large number of endogenous plasmids with unique sequences encoding unknown functions. The presence of these plasmids needs to be confirmed after each genetic manipulation. Second, the restriction modification defense systems, including that encoded by bbe02 gene lead to low transformation efficiency in B. burgdorferi. Therefore, studying the molecular basis of Lyme pathogenesis is a challenge. Furthermore, investigation of the role of a specific B. burgdorferi protein throughout infection requires a large number of mice, making it labor intensive and expensive. To overcome the problems associated with low transformation efficiency and to reduce the number of mice needed for experiments, we disrupted the bbe02 gene of a highly infectious and pathogenic B. burgdorferi strain, N40 D10/E9 through insertion of a firefly luciferase gene. The bbe02 mutant shows higher transformation efficiency and maintains luciferase activity throughout infection as detected by live imaging of mice. Infectivity and pathogenesis of this mutant were comparable to the wild-type N40 strain. This mutant will serve as an ideal parental strain to examine the roles of various B. burgdorferi proteins in Lyme pathogenesis in the mouse model in the future.

  6. Different Classes of Proteoglycans Contribute to the Attachment of Borrelia burgdorferi to Cultured Endothelial and Brain Cells

    OpenAIRE

    Leong, John M.; Wang, Hong; Magoun, Loranne; Field, Jodie A.; Morrissey, Pamela E.; Robbins, Douglas; Tatro, Jeffrey B.; Coburn, Jenifer; Parveen, Nikhat

    1998-01-01

    The Lyme disease spirochete, Borrelia burgdorferi, infects multiple tissues, such as the heart, joint, skin, and nervous system and has been shown to recognize heparan sulfate and dermatan sulfate proteoglycans. In this study, we examined the contribution of different classes of proteoglycans to the attachment of the infectious B. burgdorferi strain N40 to several immortalized cell lines and primary cultured cells, including endothelial cells and brain cells. Bacterial attachment was inhibite...

  7. High-Throughput Plasmid Content Analysis of Borrelia burgdorferi B31 by Using Luminex Multiplex Technology▿ †

    OpenAIRE

    Norris, Steven J; Howell, Jerrilyn K.; Odeh, Evelyn A.; Lin, Tao; Gao, Lihui; Diane G Edmondson

    2010-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease in North America, is an invasive pathogen that causes persistent multiorgan manifestations in humans and other mammals. Genetic studies of this bacterium are complicated by the presence of multiple plasmid replicons, many of which are readily lost during in vitro culture. The analysis of B. burgdorferi plasmid content by plasmid-specific PCR and agarose gel electrophoresis or other existing techniques is informative, but these techniqu...

  8. The Thermophilic, Homohexameric Aminopeptidase of Borrelia burgdorferi Is a Member of the M29 Family of Metallopeptidases

    OpenAIRE

    Bertin, Patrícia B.; Silene P Lozzi; Howell, Jerrilyn K.; Restrepo-Cadavid, Glória; Neves, David; Teixeira, Antonio R. L.; de Sousa, Marcelo V.; Norris, Steven J; Santana, Jaime M.

    2005-01-01

    Proteases are implicated in several aspects of the physiology of microorganisms, as well as in host-pathogen interactions. Aminopeptidases are also emerging as novel drug targets in infectious agents. In this study, we have characterized an aminopeptidase from the spirochete Borrelia burgdorferi, the causative agent of Lyme disease. The aminopeptidolytic activity was identified in cell extracts from B. burgdorferi by using the substrate leucine-7-amido-4-methylcoumarin. A protein displaying t...

  9. Expression and Immunological Analysis of the Plasmid-Borne mlp Genes of Borrelia burgdorferi Strain B31

    OpenAIRE

    Porcella, Stephen F.; Fitzpatrick, Cecily A.; Bono, James L.

    2000-01-01

    A lipoprotein gene family first identified in Borrelia burgdorferi strain 297, designated 2.9 LP and recently renamed mlp, was found on circular and linear plasmids in the genome sequence of B. burgdorferi strain B31-M1. Sequence analyses of the B31 mlp genes and physically linked variant gene families indicated that mlp gene heterogeneity is unique and unrelated to location or linkage to divergent sequences. Evidence of recombination between B31 mlp alleles was also detected. Northern blot a...

  10. Identification, Characterization, and Expression of Three New Members of the Borrelia burgdorferi Mlp (2.9) Lipoprotein Gene Family

    OpenAIRE

    Yang, Xiaofeng; Popova, Taissia G.; Hagman, Kayla E.; Wikel, Stephen K.; Schoeler, George B.; Caimano, Melissa J.; Radolf, Justin D.; Norgard, Michael V.

    1999-01-01

    We previously reported on the existence of a family of lipoprotein genes, designated 2.9 lipoprotein genes, encoded in at least seven versions on the circular (supercoiled) cp32 and cp18 plasmids of Borrelia burgdorferi 297. A distinguishing feature of the 2.9 lipoproteins were highly similar signal sequences but variable mature polypeptides that segregated into two antigenic classes. Further screenings of B. burgdorferi 297 genomic libraries led to the identification of three additional 2.9 ...

  11. Identification of endemic foci of Lyme disease: isolation of Borrelia burgdorferi from feral rodents and ticks (Dermacentor variabilis).

    OpenAIRE

    Anderson, J F; Johnson, R C; Magnarelli, L A; Hyde, F W

    1985-01-01

    Borrelia burgdorferi, the etiological agent of Lyme disease, was isolated from the blood, kidneys, spleens, eyes, or livers of white-footed mice (Peromyscus leucopus) (n = 19 of 22) and from the blood, kidneys, or spleens of eastern chipmunks (Tamias striatus) (n = 2 of 2) captured at three foci for Lyme disease in eastern Connecticut. These bacteria were cultured most frequently from spleens (n = 19) and kidneys (n = 15). B. burgdorferi persisted in one mouse for at least 60 days. One spiroc...

  12. Borrelia burgdorferi Regulates Expression of Complement Regulator-Acquiring Surface Protein 1 during the Mammal-Tick Infection Cycle

    OpenAIRE

    von Lackum, Kate; Miller, Jennifer C.; Bykowski, Tomasz; Riley, Sean P; Woodman, Michael E.; Brade, Volker; Kraiczy, Peter; Stevenson, Brian; Wallich, Reinhard

    2005-01-01

    During the natural mammal-tick infection cycle, the Lyme disease spirochete Borrelia burgdorferi comes into contact with components of the alternative complement pathway. B. burgdorferi, like many other human pathogens, has evolved the immune evasion strategy of binding two host-derived fluid-phase regulators of complement, factor H and factor H-like protein 1 (FHL-1). The borrelial complement regulator-acquiring surface protein 1 (CRASP-1) is a surface-exposed lipoprotein that binds both fac...

  13. Borrelia burgdorferi requires glycerol for maximum fitness during the tick phase of the enzootic cycle.

    Directory of Open Access Journals (Sweden)

    Christopher J Pappas

    2011-07-01

    Full Text Available Borrelia burgdorferi, the spirochetal agent of Lyme disease, is a vector-borne pathogen that cycles between a mammalian host and tick vector. This complex life cycle requires that the spirochete modulate its gene expression program to facilitate growth and maintenance in these diverse milieus. B. burgdorferi contains an operon that is predicted to encode proteins that would mediate the uptake and conversion of glycerol to dihydroxyacetone phosphate. Previous studies indicated that expression of the operon is elevated at 23°C and is repressed in the presence of the alternative sigma factor RpoS, suggesting that glycerol utilization may play an important role during the tick phase. This possibility was further explored in the current study by expression analysis and mutagenesis of glpD, a gene predicted to encode glycerol 3-phosphate dehydrogenase. Transcript levels for glpD were significantly lower in mouse joints relative to their levels in ticks. Expression of GlpD protein was repressed in an RpoS-dependent manner during growth of spirochetes within dialysis membrane chambers implanted in rat peritoneal cavities. In medium supplemented with glycerol as the principal carbohydrate, wild-type B. burgdorferi grew to a significantly higher cell density than glpD mutant spirochetes during growth in vitro at 25°C. glpD mutant spirochetes were fully infectious in mice by either needle or tick inoculation. In contrast, glpD mutants grew to significantly lower densities than wild-type B. burgdorferi in nymphal ticks and displayed a replication defect in feeding nymphs. The findings suggest that B. burgdorferi undergoes a switch in carbohydrate utilization during the mammal to tick transition. Further, the results demonstrate that the ability to utilize glycerol as a carbohydrate source for glycolysis during the tick phase of the infectious cycle is critical for maximal B. burgdorferi fitness.

  14. Borrelia burgdorferi Requires Glycerol for Maximum Fitness During The Tick Phase of the Enzootic Cycle

    Science.gov (United States)

    Pappas, Christopher J.; Iyer, Radha; Petzke, Mary M.; Caimano, Melissa J.; Radolf, Justin D.; Schwartz, Ira

    2011-01-01

    Borrelia burgdorferi, the spirochetal agent of Lyme disease, is a vector-borne pathogen that cycles between a mammalian host and tick vector. This complex life cycle requires that the spirochete modulate its gene expression program to facilitate growth and maintenance in these diverse milieus. B. burgdorferi contains an operon that is predicted to encode proteins that would mediate the uptake and conversion of glycerol to dihydroxyacetone phosphate. Previous studies indicated that expression of the operon is elevated at 23°C and is repressed in the presence of the alternative sigma factor RpoS, suggesting that glycerol utilization may play an important role during the tick phase. This possibility was further explored in the current study by expression analysis and mutagenesis of glpD, a gene predicted to encode glycerol 3-phosphate dehydrogenase. Transcript levels for glpD were significantly lower in mouse joints relative to their levels in ticks. Expression of GlpD protein was repressed in an RpoS-dependent manner during growth of spirochetes within dialysis membrane chambers implanted in rat peritoneal cavities. In medium supplemented with glycerol as the principal carbohydrate, wild-type B. burgdorferi grew to a significantly higher cell density than glpD mutant spirochetes during growth in vitro at 25°C. glpD mutant spirochetes were fully infectious in mice by either needle or tick inoculation. In contrast, glpD mutants grew to significantly lower densities than wild-type B. burgdorferi in nymphal ticks and displayed a replication defect in feeding nymphs. The findings suggest that B. burgdorferi undergoes a switch in carbohydrate utilization during the mammal to tick transition. Further, the results demonstrate that the ability to utilize glycerol as a carbohydrate source for glycolysis during the tick phase of the infectious cycle is critical for maximal B. burgdorferi fitness. PMID:21750672

  15. Increasing RpoS expression causes cell death in Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Linxu Chen

    Full Text Available RpoS, one of the two alternative σ factors in Borrelia burgdorferi, is tightly controlled by multiple regulators and, in turn, determines expression of many critical virulence factors. Here we show that increasing RpoS expression causes cell death. The immediate effect of increasing RpoS expression was to promote bacterial division and as a consequence result in a rapid increase in cell number before causing bacterial death. No DNA fragmentation or degradation was observed during this induced cell death. Cryo-electron microscopy showed induced cells first formed blebs, which were eventually released from dying cells. Apparently blebbing initiated cell disintegration leading to cell death. These findings led us to hypothesize that increasing RpoS expression triggers intracellular programs and/or pathways that cause spirochete death. The potential biological significance of induced cell death may help B. burgdorferi regulate its population to maintain its life cycle in nature.

  16. The critical role of the linear plasmid lp36 in the infectious cycle of Borrelia burgdorferi.

    Science.gov (United States)

    Jewett, Mollie W; Lawrence, Kevin; Bestor, Aaron C; Tilly, Kit; Grimm, Dorothee; Shaw, Pamela; VanRaden, Mark; Gherardini, Frank; Rosa, Patricia A

    2007-06-01

    Borrelia burgdorferi, the aetiological agent of Lyme disease, follows a life cycle that involves passage between the tick vector and the mammalian host. To investigate the role of the 36 kb linear plasmid, lp36 (also designated the B. burgdorferi K plasmid), in the infectious cycle of B. burgdorferi, we examined a clone lacking this plasmid, but containing all other plasmids known to be required for infectivity. Our results indicated that lp36 was not required for spirochete survival in the tick, but the clone lacking lp36 demonstrated low infectivity in the mammal. Restoration of lp36 to the mutant strain confirmed that the infectivity defect was due to loss of lp36. Moreover, spirochetes lacking lp36 exhibited a nearly 4-log increase in ID(50) relative to the isogenic lp36(+) clone. The infectivity defect of lp36-minus spirochetes was localized, in part, to loss of the bbk17 (adeC) gene, which encodes an adenine deaminase. This work establishes a vital role for lp36 in the infectious cycle of B. burgdorferi and identifies the bbk17 gene as a component of this plasmid that contributes to mammalian infectivity. PMID:17542926

  17. Seroprevalence of Borrelia burgdorferi antibodies in white-tailed deer from Texas.

    Science.gov (United States)

    Adetunji, Shakirat A; Krecek, Rosina C; Castellanos, Gabrielle; Morrill, John C; Blue-McLendon, Alice; Cook, Walt E; Esteve-Gassent, Maria D

    2016-08-01

    Lyme Disease is caused by the bacterial pathogen Borrelia burgdorferi, and is transmitted by the tick-vector Ixodes scapularis. It is the most prevalent arthropod-borne disease in the United States. To determine the seroprevalence of B. burgdorferi antibodies in white-tailed deer (Odocoileus virginianus) from Texas, we analyzed serum samples (n = 1493) collected during the 2001-2015 hunting seasons, using indirect ELISA. Samples with higher sero-reactivity (0.803 and above) than the negative control group (0.662) were further tested using a more specific standardized western immunoblot assay to rule out false positives. Using ELISA, 4.7% of the samples were sero-reactive against B. burgdorferi, and these originated in two eco-regions in Texas (Edwards Plateau and South Texas Plains). However, only 0.5% of the total samples were sero-reactive by standardized western immunoblot assay. Additionally, both ELISA and standardized western immunoblot assay results correlated with an increased incidence in human Lyme Disease cases reported in Texas. This is the first longitudinal study to demonstrate fluctuation in sero-reactivity of white-tailed deer to B. burgdorferi sensu stricto antigens in southern United States. Future ecological and geographical studies are needed to assess the environmental factors governing the prevalence of Lyme Disease in non-endemic areas of the southern United States. PMID:27366674

  18. Live Attenuated Borrelia burgdorferi Targeted Mutants in an Infectious Strain Background Protect Mice from Challenge Infection.

    Science.gov (United States)

    Hahn, Beth L; Padmore, Lavinia J; Ristow, Laura C; Curtis, Michael W; Coburn, Jenifer

    2016-08-01

    Borrelia burgdorferi, B. garinii, and B. afzelii are all agents of Lyme disease in different geographic locations. If left untreated, Lyme disease can cause significant and long-term morbidity, which may continue after appropriate antibiotic therapy has been administered and live bacteria are no longer detectable. The increasing incidence and geographic spread of Lyme disease are renewing interest in the vaccination of at-risk populations. We took the approach of vaccinating mice with two targeted mutant strains of B. burgdorferi that, unlike the parental strain, are avirulent in mice. Mice vaccinated with both strains were protected against a challenge with the parental strain and a heterologous B. burgdorferi strain by either needle inoculation or tick bite. In ticks, the homologous strain was eliminated but the heterologous strain was not, suggesting that the vaccines generated a response to antigens that are produced by the bacteria both early in mammalian infection and in the tick. Partial protection against B. garinii infection was also conferred. Protection was antibody mediated, and reactivity to a variety of proteins was observed. These experiments suggest that live attenuated B. burgdorferi strains may be informative regarding the identification of protective antigens produced by the bacteria and recognized by the mouse immune system in vivo Further work may illuminate new candidates that are effective and safe for the development of Lyme disease vaccines. PMID:27335385

  19. Nod2 suppresses Borrelia burgdorferi mediated murine Lyme arthritis and carditis through the induction of tolerance.

    Directory of Open Access Journals (Sweden)

    Tanja Petnicki-Ocwieja

    Full Text Available The internalization of Borrelia burgdorferi, the causative agent of Lyme disease, by phagocytes is essential for an effective activation of the immune response to this pathogen. The intracellular, cytosolic receptor Nod2 has been shown to play varying roles in either enhancing or attenuating inflammation in response to different infectious agents. We examined the role of Nod2 in responses to B. burgdorferi. In vitro stimulation of Nod2 deficient bone marrow derived macrophages (BMDM resulted in decreased induction of multiple cytokines, interferons and interferon regulated genes compared with wild-type cells. However, B. burgdorferi infection of Nod2 deficient mice resulted in increased rather than decreased arthritis and carditis compared to control mice. We explored multiple potential mechanisms for the paradoxical response in in vivo versus in vitro systems and found that prolonged stimulation with a Nod2 ligand, muramyl dipeptide (MDP, resulted in tolerance to stimulation by B. burgdorferi. This tolerance was lost with stimulation of Nod2 deficient cells that cannot respond to MDP. Cytokine patterns in the tolerance model closely paralleled cytokine profiles in infected Nod2 deficient mice. We propose a model where Nod2 has an enhancing role in activating inflammation in early infection, but moderates inflammation after prolonged exposure to the organism through induction of tolerance.

  20. BB0172, a Borrelia burgdorferi outer membrane protein that binds integrin α3β1.

    Science.gov (United States)

    Wood, Elaine; Tamborero, Silvia; Mingarro, Ismael; Esteve-Gassent, Maria D

    2013-08-01

    Lyme disease is a multisystemic disorder caused by Borrelia burgdorferi infection. Upon infection, some B. burgdorferi genes are upregulated, including members of the microbial surface components recognizing adhesive matrix molecule (MSCRAMM) protein family, which facilitate B. burgdorferi adherence to extracellular matrix components of the host. Comparative genome analysis has revealed a new family of B. burgdorferi proteins containing the von Willebrand factor A (vWFA) domain. In the present study, we characterized the expression and membrane association of the vWFA domain-containing protein BB0172 by using in vitro transcription/translation systems in the presence of microsomal membranes and with detergent phase separation assays. Our results showed evidence of BB0172 localization in the outer membrane, the orientation of the vWFA domain to the extracellular environment, and its function as a metal ion-dependent integrin-binding protein. This is the first report of a borrelial adhesin with a metal ion-dependent adhesion site (MIDAS) motif that is similar to those observed in eukaryotic integrins and has a similar function. PMID:23687274

  1. In Vitro Activities of the Everninomicin SCH 27899 and Other Newer Antimicrobial Agents against Borrelia burgdorferi

    OpenAIRE

    Lisa L. Dever; Torigian, Christine V.; Barbour, Alan G.

    1999-01-01

    The in vitro activity of the everninomicin antibiotic SCH 27899 against 17 isolates of Borrelia spp. was investigated. MICs ranged from 0.06 to 0.5 μg/ml. Time-kill studies with the B31 strain of B. burgdorferi demonstrated ≥3-log10-unit killing after 72 h with concentrations representing four times the MIC. The in vitro activity of four other newer antimicrobial agents, meropenem, cefepime, quinupristin-dalfopristin, and linezolid, was also tested against the B31 strain. Meropenem was the mo...

  2. Développement de Borrelia burgdorferi dans la tique Ixodes ricinus

    OpenAIRE

    Gern, Lise; Z. Zhu; Aeschlimann, André

    2011-01-01

    Nous avons étudié le développement de Borrelia burgdorferi dans son vecteur européen, la tique Ixodes ricinus, lors du repas sanguin. Nos résultats montrent que durant la phase de gorgement lente, les spirochètes présents dans l'intestin se multiplient. Chez certains individus, quelques spirochètes traversent la paroi intestinale et produisent une infection systémique de la tique. La présence de borrélies dans les conduits des glandes salivaires semblent prouver leur transmission via la saliv...

  3. Statistical analysis of the distribution of amino acids in Borrelia burgdorferi genome under different genetic codes

    Science.gov (United States)

    García, José A.; Alvarez, Samantha; Flores, Alejandro; Govezensky, Tzipe; Bobadilla, Juan R.; José, Marco V.

    2004-10-01

    The genetic code is considered to be universal. In order to test if some statistical properties of the coding bacterial genome were due to inherent properties of the genetic code, we compared the autocorrelation function, the scaling properties and the maximum entropy of the distribution of distances of amino acids in sequences obtained by translating protein-coding regions from the genome of Borrelia burgdorferi, under different genetic codes. Overall our results indicate that these properties are very stable to perturbations made by altering the genetic code. We also discuss the evolutionary likely implications of the present results.

  4. Evidencias de infección por Borrelia burgdorferi sensu lato en Cuba

    OpenAIRE

    Rodríguez González, Islay

    2012-01-01

    Se presentan los resultados de un conjunto de investigaciones realizadas con el propósito de aportar evidencias científicas sobre la presencia de la infección por Borrelia burgdorferi sensu lato en Cuba, que incluyó la evaluación de métodos microbiológicos para la detección de esta espiroqueta, la confirmación de la infección en muestras clínicas de pacientes con sospechas clínico-epidemiológicas de enfermedad de Lyme, la estimación de la seroprevalencia de anticuerpos contra este agent...

  5. A new (14th) member of Borrelia burgdorferi sensu lato complex

    Czech Academy of Sciences Publication Activity Database

    Oliver, J.; Rudenko, Natalia; Golovchenko, Maryna; Grubhoffer, Libor

    2008-01-01

    Roč. 14, S7 (2008), S110-S110. ISSN 1198-743X. [ECCMID /18./. 19.04.2008-22.04.2008, Barcelona] R&D Projects: GA ČR(CZ) GA524/06/1479; GA MŠk(CZ) LC06009 Grant ostatní: National Institutes of Health (NIH)(US) R37AI-24899; Centers for Disease Control and Prevention (CDC)(US) U50/CCU410282 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia burgdorferi s.l. complex * new member * multilocus sequence analysis Subject RIV: EC - Immunology

  6. Geographical differences in seroprevalence of Borrelia burgdorferi antibodies in Norway, 2011-2013.

    Science.gov (United States)

    Vestrheim, Didrik F; White, Richard A; Aaberge, Ingeborg S; Aase, Audun

    2016-07-01

    Detection of specific antibodies against Borrelia burgdorferi sensu lato is a useful aid for the diagnosis of Lyme borreliosis. However, antibodies are present in the general population. The seroprevalence increase with age, and varies according to the prevalence of infected ticks. We performed a seroprevalence study of IgM and IgG antibody reactivity against B. burgdorferi sensu lato in Norway by age-groups and geography, in order to provide a reference set of seroprevalence to inform the interpretation of positive test results. We used two commercially available enzyme immuno assays (EIA) and a multiplexed bead assay to detect Borrelia IgG antibodies in a convenience sample of 3057 sera collected from clinical chemistry laboratories in 10 of 19 counties in Norway between December 2011 and January 2013. We estimated seroprevalence by age and county by a logistic regression model. IgM antibodies were detected by two commercially available EIAs and a multiplexed bead assay. The overall seroprevalence of Borrelia IgG was 4.0% (95% CI: 2.4-6.6%) and 4.2% (2.6-6.8%) by the two EIAs, respectively. The seroprevalence increased by age, and by geography from north to south. The IgG assays showed a good agreement for positive test results. All sera positive for IgG in the multiplexed bead assay reacted with the VlsE antigen, and also had high antibody levels by EIA. The Borrelia seroprevalence varied by geography and increased by age. The results indicate regional differences in pre-test probabilities for positive test results, and can inform the interpretation of laboratory results. PMID:26961275

  7. Multiple infections of Ixodes scapularis ticks by Borrelia burgdorferi as revealed by single-strand conformation polymorphism analysis.

    OpenAIRE

    Guttman, D. S.; Wang, P. W.; Wang, I. N.; Bosler, E. M.; Luft, B J; Dykhuizen, D E

    1996-01-01

    The genetic heterogeneity of the spirochete Borrelia burgdorferi within single adult black-legged ticks from Shelter Island, N.Y., was determined by cold, single-strand conformation polymorphism (SSCP) analysis. The central region of the ospA gene of B. burgdorferi from infected ticks was amplified by nested PCR. Amplified product of the correct size was obtained from 20 to 45 ticks (44%). This is the fraction of ticks that is expected to be infected with B. burgdorferi. Four variant classes ...

  8. A Borrelia burgdorferi Surface-Exposed Transmembrane Protein Lacking Detectable Immune Responses Supports Pathogen Persistence and Constitutes a Vaccine Target.

    Science.gov (United States)

    Kung, Faith; Kaur, Simarjot; Smith, Alexis A; Yang, Xiuli; Wilder, Cara N; Sharma, Kavita; Buyuktanir, Ozlem; Pal, Utpal

    2016-06-01

    Borrelia burgdorferi harbors a limited set of transmembrane surface proteins, most of which constitute key targets of humoral immune responses. Here we show that BB0405, a conserved membrane-spanning protein of unknown function, fails to evoke detectable antibody responses despite its extracellular exposure. bb0405 is a member of an operon and ubiquitously expressed throughout the rodent-tick infection cycle. The gene product serves an essential function in vivo, as bb0405-deletion mutants are unable to transmit from ticks and establish infection in mammalian hosts. Despite the lack of BB0405-specific immunoglobulin M or immunoglobulin G antibodies during natural infection, mice immunized with a recombinant version of the protein elicited high-titer and remarkably long-lasting antibody responses, conferring significant host protection against tick-borne infection. Taken together, these studies highlight the essential role of an apparently immune-invisible borrelial transmembrane protein in facilitating infection and its usefulness as a target of protective host immunity blocking the transmission of B. burgdorferi. PMID:26747708

  9. An Enhanced ELISPOT Assay for Sensitive Detection of Antigen-Specific T Cell Responses to Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Gottfried H. Kellermann

    2013-09-01

    Full Text Available Lyme Borreliosis is an infectious disease caused by the spirochete Borrelia burgdorferi that is transmitted through the bite of infected ticks. Both B cell-mediated humoral immunity and T cell immunity develop during natural Borrelia infection. However, compared with humoral immunity, the T cell response to Borrelia infection has not been well elucidated. In this study, a novel T cell-based assay was developed and validated for the sensitive detection of antigen-specific T cell response to B. burgdorferi. Using interferon-g as a biomarker, we developed a new enzyme-linked immunospot method (iSpot Lyme™ to detect Borrelia antigen-specific effector/memory T cells that were activated in vivo by exposing them to recombinant Borrelia antigens ex vivo. To test this new method as a potential laboratory diagnostic tool, we performed a clinical study with a cohort of Borrelia positive patients and healthy controls. We demonstrated that the iSpot Lyme assay has a significantly higher specificity and sensitivity compared with the Western Blot assay that is currently used as a diagnostic measure. A comprehensive evaluation of the T cell response to Borrelia infection should, therefore, provide new insights into the pathogenesis, diagnosis, treatment and monitoring of Lyme disease.

  10. NKT cells prevent chronic joint inflammation after infection with Borrelia burgdorferi.

    Science.gov (United States)

    Tupin, Emmanuel; Benhnia, Mohammed Rafii-El-Idrissi; Kinjo, Yuki; Patsey, Rebeca; Lena, Christopher J; Haller, Matthew C; Caimano, Melissa J; Imamura, Masakazu; Wong, Chi-Huey; Crotty, Shane; Radolf, Justin D; Sellati, Timothy J; Kronenberg, Mitchell

    2008-12-16

    Borrelia burgdorferi is the etiologic agent of Lyme disease, a multisystem inflammatory disorder that principally targets the skin, joints, heart, and nervous system. The role of T lymphocytes in the development of chronic inflammation resulting from B. burgdorferi infection has been controversial. We previously showed that natural killer T (NKT) cells with an invariant (i) TCR alpha chain (iNKT cells) recognize glycolipids from B. burgdorferi, but did not establish an in vivo role for iNKT cells in Lyme disease pathogenesis. Here, we evaluate the importance of iNKT cells for host defense against these pathogenic spirochetes by using Valpha14i NKT cell-deficient (Jalpha18(-/-)) BALB/c mice. On tick inoculation with B. burgdorferi, Jalpha18(-/-) mice exhibited more severe and prolonged arthritis as well as a reduced ability to clear spirochetes from infected tissues. Valpha14i NKT cell deficiency also resulted in increased production of antibodies directed against both B. burgdorferi protein antigens and borrelial diacylglycerols; the latter finding demonstrates that anti-glycolipid antibody production does not require cognate help from Valpha14i NKT cells. Valpha14i NKT cells in infected wild-type mice expressed surface activation markers and produced IFNgamma in vivo after infection, suggesting a participatory role for this unique population in cellular immunity. Our data are consistent with the hypothesis that the antigen-specific activation of Valpha14i NKT cells is important for the prevention of persistent joint inflammation and spirochete clearance, and they counter the long-standing notion that humoral rather than cellular immunity is sufficient to facilitate Lyme disease resolution. PMID:19060201

  11. Evaluation of the importance of VlsE antigenic variation for the enzootic cycle of borrelia burgdorferi

    Science.gov (United States)

    Efficient acquisition and transmission of Borrelia burgdorferi by the tick vector, and the ability to persistently infect both vector and host, are important elements for the life cycle of the Lyme disease pathogen. Previous work has provided strong evidence implicating the significance of the vls l...

  12. Seroprevalence of Borrelia burgdorferi sensu lato and tick-borne encephalitis virus in zoo animal species in the Czech Republic

    Czech Academy of Sciences Publication Activity Database

    Širmarová, J.; Tichá, L.; Golovchenko, Maryna; Salát, Jiří; Grubhoffer, L.; Rudenko, Natalia; Nowotny, N.; Růžek, Daniel

    2014-01-01

    Roč. 5, č. 5 (2014), s. 523-527. ISSN 1877-959X R&D Projects: GA ČR GAP502/11/2116 Institutional support: RVO:60077344 Keywords : Tick-borne encephalitis virus * Borrelia burgdorferi sensu lato * Lyme borreliosis * Seroprevalence * Zoo animals Subject RIV: EE - Microbiology, Virology Impact factor: 2.718, year: 2014

  13. Are the specialized bird ticks, Ixodes arboricola and I. frontalis, competent vectors for Borrelia burgdorferi sensu lato?

    NARCIS (Netherlands)

    Heylen, D.; Sprong, H.; Van Oers, K.; Fonville, M.; Matthysen, E.

    2014-01-01

    Our study tested whether two European bird-specialized ticks, Ixodes arboricola and I. frontalis, can act as vectors in the transmission cycles of Borrelia burgdorferi s.l. The ticks have contrasting ecologies but share songbird hosts (such as the great tit, Parus major) with the generalist I. ricin

  14. Are the specialized bird ticks, Ixodes arboricola and I. frontalis, competent vectors for Borrelia burgdorferi sensu lato?

    NARCIS (Netherlands)

    Heylen, D.; Sprong, H.; Oers, van K.; Fonville, M.; Leirs, H.; Matthysen, E.

    2014-01-01

    Our study tested whether two European bird-specialized ticks, Ixodes arboricola and I. frontalis, can act as vectors in the transmission cycles of Borrelia burgdorferi s.l. The ticks have contrasting ecologies but share songbird hosts (such as the great tit, Parus major) with the generalist I. ricin

  15. Fluorescent membrane markers elucidate the association of Borrelia burgdorferi with tick cell lines.

    Science.gov (United States)

    Teixeira, R C; Baêta, B A; Ferreira, J S; Medeiros, R C; Maya-Monteiro, C M; Lara, F A; Bell-Sakyi, L; Fonseca, A H

    2016-06-20

    This study aimed to describe the association of Borrelia burgdorferi s.s. with ixodid tick cell lines by flow cytometry and fluorescence and confocal microscopy. Spirochetes were stained with a fluorescent membrane marker (PKH67 or PKH26), inoculated into 8 different tick cell lines and incubated at 30°C for 24 h. PKH efficiently stained B. burgdorferi without affecting bacterial viability or motility. Among the tick cell lines tested, the Rhipicephalus appendiculatus cell line RA243 achieved the highest percentage of association/internalization, with both high (90%) and low (10%) concentrations of BSK-H medium in tick cell culture medium. Treatment with cytochalasin D dramatically reduced the average percentage of cells with internalized spirochetes, which passed through a dramatic morphological change during their internalization by the host cell as observed in time-lapse photography. Almost all of the fluorescent bacteria were seen to be inside the tick cells. PKH labeling of borreliae proved to be a reliable and valuable tool to analyze the association of spirochetes with host cells by flow cytometry, confocal and fluorescence microscopy. PMID:27332772

  16. Genome stability of Lyme disease spirochetes: comparative genomics of Borrelia burgdorferi plasmids.

    Directory of Open Access Journals (Sweden)

    Sherwood R Casjens

    Full Text Available Lyme disease is the most common tick-borne human illness in North America. In order to understand the molecular pathogenesis, natural diversity, population structure and epizootic spread of the North American Lyme agent, Borrelia burgdorferi sensu stricto, a much better understanding of the natural diversity of its genome will be required. Towards this end we present a comparative analysis of the nucleotide sequences of the numerous plasmids of B. burgdorferi isolates B31, N40, JD1 and 297. These strains were chosen because they include the three most commonly studied laboratory strains, and because they represent different major genetic lineages and so are informative regarding the genetic diversity and evolution of this organism. A unique feature of Borrelia genomes is that they carry a large number of linear and circular plasmids, and this work shows that strains N40, JD1, 297 and B31 carry related but non-identical sets of 16, 20, 19 and 21 plasmids, respectively, that comprise 33-40% of their genomes. We deduce that there are at least 28 plasmid compatibility types among the four strains. The B. burgdorferi ∼900 Kbp linear chromosomes are evolutionarily exceptionally stable, except for a short ≤20 Kbp plasmid-like section at the right end. A few of the plasmids, including the linear lp54 and circular cp26, are also very stable. We show here that the other plasmids, especially the linear ones, are considerably more variable. Nearly all of the linear plasmids have undergone one or more substantial inter-plasmid rearrangements since their last common ancestor. In spite of these rearrangements and differences in plasmid contents, the overall gene complement of the different isolates has remained relatively constant.

  17. Genome Stability of Lyme Disease Spirochetes: Comparative Genomics of Borrelia burgdorferi Plasmids

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    Casjens S. R.; Dunn J.; Mongodin, E. F.; Qiu, W.-G.; Luft, B. J.; Schutzer, S. E.; Gilcrease, E. B.; Huang, W. M.; Vujadinovic, M.; Aron, J. K.; Vargas, L. C.; Freeman, S.; Radune, D.; Weidman, J. F.; Dimitrov, G. I.; Khouri, H. M.; Sosa, J. E.; Halpin, R. A.; Fraser, C. M.

    2012-03-14

    Lyme disease is the most common tick-borne human illness in North America. In order to understand the molecular pathogenesis, natural diversity, population structure and epizootic spread of the North American Lyme agent, Borrelia burgdorferi sensu stricto, a much better understanding of the natural diversity of its genome will be required. Towards this end we present a comparative analysis of the nucleotide sequences of the numerous plasmids of B. burgdorferi isolates B31, N40, JD1 and 297. These strains were chosen because they include the three most commonly studied laboratory strains, and because they represent different major genetic lineages and so are informative regarding the genetic diversity and evolution of this organism. A unique feature of Borrelia genomes is that they carry a large number of linear and circular plasmids, and this work shows that strains N40, JD1, 297 and B31 carry related but non-identical sets of 16, 20, 19 and 21 plasmids, respectively, that comprise 33-40% of their genomes. We deduce that there are at least 28 plasmid compatibility types among the four strains. The B. burgdorferi {approx}900 Kbp linear chromosomes are evolutionarily exceptionally stable, except for a short {le}20 Kbp plasmid-like section at the right end. A few of the plasmids, including the linear lp54 and circular cp26, are also very stable. We show here that the other plasmids, especially the linear ones, are considerably more variable. Nearly all of the linear plasmids have undergone one or more substantial inter-plasmid rearrangements since their last common ancestor. In spite of these rearrangements and differences in plasmid contents, the overall gene complement of the different isolates has remained relatively constant.

  18. Temperature-induced regulation of RpoS by a small RNA in Borrelia burgdorferi.

    Science.gov (United States)

    Lybecker, Meghan C; Samuels, D Scott

    2007-05-01

    The alternative sigma factor RpoS (sigma38 or sigmaS) plays a central role in the reciprocal regulation of the virulence-associated major outer surface proteins OspC and OspA in Borrelia burgdorferi, the Lyme disease spirochete. Temperature is one of the key environmental signals controlling RpoS, but the molecular mechanism by which the signal is transduced remains unknown. Herein, we identify and describe a small non-coding RNA, DsrABb, that regulates the temperature-induced increase in RpoS. A novel 5' end of the rpoS mRNA was identified and DsrABb has the potential to extensively base-pair with the upstream region of this rpoS transcript. We demonstrate that B. burgdorferi strains lacking DsrABb do not upregulate RpoS and OspC in response to an increase in temperature, but do regulate RpoS and OspC in response to changes in pH and cell density. Analyses of the rpoS and ospC steady-state mRNA levels in the dsrABb mutant indicate that DsrABb regulates RpoS post-transcriptionally. The 5' and 3' ends of DsrABb were mapped, demonstrating that at least four species exist with sizes ranging from 213 to 352 nucleotides. We hypothesize that DsrABb binds to the upstream region of the rpoS mRNA and stimulates translation by releasing the Shine-Dalgarno sequence and start site from a stable secondary structure. Therefore, we postulate that DsrABb is a molecular thermometer regulating RpoS in Borrelia burgdorferi. PMID:17501929

  19. Functional analysis of the Borrelia burgdorferi bba64 gene product in murine infection via tick infestation.

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    Toni G Patton

    Full Text Available Borrelia burgdorferi, the causative agent of Lyme borreliosis, is transmitted to humans from the bite of Ixodes spp. ticks. During the borrelial tick-to-mammal life cycle, B. burgdorferi must adapt to many environmental changes by regulating several genes, including bba64. Our laboratory recently demonstrated that the bba64 gene product is necessary for mouse infectivity when B. burgdorferi is transmitted by an infected tick bite, but not via needle inoculation. In this study we investigated the phenotypic properties of a bba64 mutant strain, including 1 replication during tick engorgement, 2 migration into the nymphal salivary glands, 3 host transmission, and 4 susceptibility to the MyD88-dependent innate immune response. Results revealed that the bba64 mutant's attenuated infectivity by tick bite was not due to a growth defect inside an actively feeding nymphal tick, or failure to invade the salivary glands. These findings suggested there was either a lack of spirochete transmission to the host dermis or increased susceptibility to the host's innate immune response. Further experiments showed the bba64 mutant was not culturable from mouse skin taken at the nymphal bite site and was unable to establish infection in MyD88-deficient mice via tick infestation. Collectively, the results of this study indicate that BBA64 functions at the salivary gland-to-host delivery interface of vector transmission and is not involved in resistance to MyD88-mediated innate immunity.

  20. Mechanisms generating long range correlation in nucleotide composition of the Borrelia Burgdorferi genome

    Science.gov (United States)

    Mackiewicz, P.; Gierlik, A.; Kowalczuk, M.; Szczepanik, D.; Dudek, M. R.; Cebrat, S.

    1999-12-01

    We have analysed protein coding and intergenic sequences in the Borrelia burgdorferi (the Lyme disease bacterium) genome using different kinds of DNA walks. Genes occupying the leading strand of DNA have significantly different nucleotide composition from genes occupying the lagging strand. Nucleotide compositional bias of the two DNA strands reflects the aminoacid composition of proteins. 96% of genes coding for ribosomal proteins lie on the leading DNA strand, which suggests that the positions of these as well as other genes are non-random. In the B. burgdorferi genome, the asymmetry in intergenic DNA sequences is lower than the asymmetry in the third positions in codons. All these characters of the B. burgdorferi genome suggest that both replication-associated mutational pressure and recombination mechanisms have established the specific structure of the genome and now any recombination leading to inversion of a gene in respect to the direction of replication is forbidden. This property of the genome allows us to assume that it is in a steady state, which enables us to fix some parameters for simulations of DNA evolution.

  1. The BBA33 lipoprotein binds collagen and impacts Borrelia burgdorferi pathogenesis

    Science.gov (United States)

    Zhi, Hui; Weening, Eric H.; Barbu, Elena Magda; Hyde, Jenny A.; Höök, Magnus; Skare, Jon T.

    2016-01-01

    Summary Borrelia burgdorferi , the etiologic agent of Lyme disease, adapts to the mammalian hosts by differentially expressing several genes in the BosR and Rrp2-RpoN-RpoS dependent pathways, resulting in a distinct protein profile relative to that seen for survival in the Ixodes spp. tick. Previous studies indicate that a putative lipoprotein, BBA33, is produced in an RpoS-dependent manner under conditions that mimic the mammalian component of the borrelial lifecycle. However, the significance and function for BBA33 is not known. Given its linkage to the BosR/Rrp2-RpoN-RpoS regulatory cascade, we hypothesized that BBA33 facilitates B. burgdorferi infection in the mammalian host. The deletion of bba33 eliminated B. burgdorferi infectivity in C3H mice, which was rescued by genetic complementation with intact bba33. With regard to function, a combinatorial peptide approach, coupled with subsequent in vitro binding assays, indicated that BBA33 binds to collagen type VI and, to a lesser extent, collagen type IV. Whole cell binding assays demonstrated BBA33-dependent binding to human collagen type VI. Taken together, these results suggest that BBA33 interacts with collagenous structures and may function as an adhesin in a process that is required to prevent bacterial clearance. PMID:25560615

  2. Lyme disease and the detection of Borrelia burgdorferi genospecies in Ixodes ricinus ticks from central Italy

    Directory of Open Access Journals (Sweden)

    Ilaria Pascucci

    2010-06-01

    Full Text Available The Province of Pesaro-Urbino, situated in the Marche Region of central Italy, can be considered to be an area at risk for Lyme disease because of its ecological features. Field data are not yet available although the disease is known to be present in neighbouring areas. During a field study lasting twelve months, ticks were collected from the vegetation, from wild cervids and also from humans who reported a tick bite at the local hospital. All ticks were identified and Ixodes ricinus specimens were tested using three different polymerase chain reaction tests for the detection of Borrelia burgdorferi sensu lato (sl. To identify the genospecies of B. burgdorferi sl, a fragment of the 5S-23S ribosomal rRNA intergenic spacer of the positive samples was amplified and then sequenced. Sequencing of the 5S-23S intergenic spacer led to the identification of two different genospecies, namely: B. burgdorferi sensu stricto and B. lusitaniae, both of which are involved in cases of human infection. Findings on the host-tick relationships and on the genospecies involved in the cycle of borreliosis confirm the suitable conditions for Lyme disease in the study area. The results concur with previous findings reported in the Mediterranean region.

  3. Novel methods for surveying reservoir hosts and vectors of Borrelia burgdorferi in Northern Minnesota

    Science.gov (United States)

    Seifert, Veronica Aili

    Lyme disease is the most prevalent tick-borne disease in North America and presents challenges to clinicians, researchers and the public in diagnosis, treatment and prevention. Lyme disease is caused by the spirochete, Borrelia burgdorferi, which is a zoonotic pathogen obligate upon hematophagous arthropod vectors and propagates in small mammal reservoir hosts. Identifying factors governing zoonotic diseases within regions of high-risk provides local health and agricultural agencies with necessary information to formulate public policy and implement treatment protocols to abate the rise and expansion of infectious disease outbreaks. In the United States, the documented primary reservoir host of Lyme disease is the white-footed mouse, Peromyscus leucopus, and the arthropod vector is the deer tick, Ixodes scapularis. Reducing the impact of Lyme disease will need novel methods for identifying both the reservoir host and the tick vector. The reservoir host, Peromyscus leucopus is difficult to distinguish from the virtually identical Peromyscus maniculatus that also is present in Northern Minnesota, a region where Lyme disease is endemic. Collection of the Ixodes tick, the Lyme disease vector, is difficult as this is season dependent and differs from year to year. This study develops new strategies to assess the extent of Borrelia burgdorferi in the local environment of Northern Minnesota. A selective and precise method to identify Peromyscus species was developed. This assay provides a reliable and definitive method to identify the reservoir host, Peromyscus leucopus from a physically identical and sympatric Peromyscus species, Peromyscus maniculatus. A new strategy to collect ticks for measuring the disbursement of Borrelia was employed. Students from local high schools were recruited to collect ticks. This strategy increased the available manpower to cover greater terrain, provided students with valuable experience in research methodology, and highlighted the

  4. Genotypic diversity of an emergent population of Borrelia burgdorferi at a coastal Maine island recently colonized by Ixodes scapularis.

    Science.gov (United States)

    MacQueen, Douglas D; Lubelczyk, Charles; Elias, Susan P; Cahill, Bruce K; Mathers, Amy J; Lacombe, Eleanor H; Rand, Peter W; Smith, Robert P

    2012-06-01

    The recent range expansion of Ixodes scapularis has been accompanied by the emergence of Borrelia burgdorferi. The development of genetic diversity in B. burgdorferi at these sites of emergence and its relationship to range expansion is poorly understood. We followed colonization of I. scapularis on a coastal Maine island over a 17-year period. B. burgdorferi's emergence was documented, as was expansion of ospC strain diversity. Ticks collected from rodents and vegetation were examined for the presence of B. burgdorferi. Sequencing and reverse line blot were used to detect B. burgdorferi ospC major groups (oMG). No I. scapularis were found until year four of the study, after which time they increased in abundance. No B. burgdorferi was detected by darkfield microscopy in I. scapularis until 10 years into the study, when 4% of adult ticks were infected. Seven years later, 43% of adult ticks were infected. In 2003, one oMG accounted for 91% of B. burgdorferi strains. This "founder" strain persisted in 2005, but by 2007 was a minority of the 7 oMGs present. Given the island's isolation, gene flow by avian introduction of multiple strains is suggested in the development of B. burgdorferi oMG diversity. PMID:22217172

  5. Linkages of Weather and Climate With Ixodes scapularis and Ixodes pacificus (Acari: Ixodidae), Enzootic Transmission of Borrelia burgdorferi, and Lyme Disease in North America.

    Science.gov (United States)

    Eisen, Rebecca J; Eisen, Lars; Ogden, Nicholas H; Beard, Charles B

    2016-03-01

    Lyme disease has increased both in incidence and geographic extent in the United States and Canada over the past two decades. One of the underlying causes is changes during the same time period in the distribution and abundance of the primary vectors: Ixodes scapularis Say and Ixodes pacificus Cooley and Kohls in eastern and western North America, respectively. Aside from short periods of time when they are feeding on hosts, these ticks exist in the environment where temperature and relative humidity directly affect their development, survival, and host-seeking behavior. Other important factors that strongly influence tick abundance as well as the proportion of ticks infected with the Lyme disease spirochete, Borrelia burgdorferi, include the abundance of hosts for the ticks and the capacity of tick hosts to serve as B. burgdorferi reservoirs. Here, we explore the linkages between climate variation and: 1) duration of the seasonal period and the timing of peak activity; 2) geographic tick distributions and local abundance; 3) enzootic B. burgdorferi transmission cycles; and 4) Lyme disease cases. We conclude that meteorological variables are most influential in determining host-seeking phenology and development, but, while remaining important cofactors, additional variables become critical when exploring geographic distribution and local abundance of ticks, enzootic transmission of B. burgdorferi, and Lyme disease case occurrence. Finally, we review climate change-driven projections for future impact on vector ticks and Lyme disease and discuss knowledge gaps and research needs. PMID:26681789

  6. Use of an endogenous plasmid locus for stable in trans complementation in Borrelia burgdorferi.

    Science.gov (United States)

    Kasumba, Irene N; Bestor, Aaron; Tilly, Kit; Rosa, Patricia A

    2015-02-01

    Targeted mutagenesis and complementation are important tools for studying genes of unknown function in the Lyme disease spirochete Borrelia burgdorferi. A standard method of complementation is reintroduction of a wild-type copy of the targeted gene on a shuttle vector. However, shuttle vectors are present at higher copy numbers than B. burgdorferi plasmids and are potentially unstable in the absence of selection, thereby complicating analyses in the mouse-tick infectious cycle. B. burgdorferi has over 20 plasmids, with some, such as linear plasmid 25 (lp25), carrying genes required by the spirochete in vivo but relatively unstable during in vitro cultivation. We propose that complementation on an endogenous plasmid such as lp25 would overcome the copy number and in vivo stability issues of shuttle vectors. In addition, insertion of a selectable marker on lp25 could ensure its stable maintenance by spirochetes in culture. Here, we describe the construction of a multipurpose allelic-exchange vector containing a multiple-cloning site and either of two selectable markers. This suicide vector directs insertion of the complementing gene into the bbe02 locus, a site on lp25 that was previously shown to be nonessential during both in vitro and in vivo growth. We demonstrate the functional utility of this strategy by restoring infectivity to an ospC mutant through complementation at this site on lp25 and stable maintenance of the ospC gene throughout mouse infection. We conclude that this represents a convenient and widely applicable method for stable gene complementation in B. burgdorferi. PMID:25452278

  7. Identification of new drug candidates against Borrelia burgdorferi using high-throughput screening.

    Science.gov (United States)

    Pothineni, Venkata Raveendra; Wagh, Dhananjay; Babar, Mustafeez Mujtaba; Inayathullah, Mohammed; Solow-Cordero, David; Kim, Kwang-Min; Samineni, Aneesh V; Parekh, Mansi B; Tayebi, Lobat; Rajadas, Jayakumar

    2016-01-01

    Lyme disease is the most common zoonotic bacterial disease in North America. It is estimated that >300,000 cases per annum are reported in USA alone. A total of 10%-20% of patients who have been treated with antibiotic therapy report the recrudescence of symptoms, such as muscle and joint pain, psychosocial and cognitive difficulties, and generalized fatigue. This condition is referred to as posttreatment Lyme disease syndrome. While there is no evidence for the presence of viable infectious organisms in individuals with posttreatment Lyme disease syndrome, some researchers found surviving Borrelia burgdorferi population in rodents and primates even after antibiotic treatment. Although such observations need more ratification, there is unmet need for developing the therapeutic agents that focus on removing the persisting bacterial form of B. burgdorferi in rodent and nonhuman primates. For this purpose, high-throughput screening was done using BacTiter-Glo assay for four compound libraries to identify candidates that stop the growth of B. burgdorferi in vitro. The four chemical libraries containing 4,366 compounds (80% Food and Drug Administration [FDA] approved) that were screened are Library of Pharmacologically Active Compounds (LOPAC1280), the National Institutes of Health Clinical Collection, the Microsource Spectrum, and the Biomol FDA. We subsequently identified 150 unique compounds, which inhibited >90% of B. burgdorferi growth at a concentration of <25 µM. These 150 unique compounds comprise many safe antibiotics, chemical compounds, and also small molecules from plant sources. Of the 150 unique compounds, 101 compounds are FDA approved. We selected the top 20 FDA-approved molecules based on safety and potency and studied their minimum inhibitory concentration and minimum bactericidal concentration. The promising safe FDA-approved candidates that show low minimum inhibitory concentration and minimum bactericidal concentration values can be chosen as lead

  8. Evaluation of in-vitro antibiotic susceptibility of different morphological forms of Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Sapi E

    2011-05-01

    Full Text Available Eva Sapi1, Navroop Kaur1, Samuel Anyanwu1, David F Luecke1, Akshita Datar1, Seema Patel1, Michael Rossi1, Raphael B Stricker21Lyme Disease Research Group, Department of Biology and Environmental Sciences, University of New Haven, New Haven, CT, USA; 2International Lyme and Associated Diseases Society, Bethesda, MD, USABackground: Lyme disease is a tick-borne illness caused by the spirochete Borrelia burgdorferi. Although antibiotic therapy is usually effective early in the disease, relapse may occur when administration of antibiotics is discontinued. Studies have suggested that resistance and recurrence of Lyme disease might be due to formation of different morphological forms of B. burgdorferi, namely round bodies (cysts and biofilm-like colonies. Better understanding of the effect of antibiotics on all morphological forms of B. burgdorferi is therefore crucial to provide effective therapy for Lyme disease.Methods: Three morphological forms of B. burgdorferi (spirochetes, round bodies, and biofilm-like colonies were generated using novel culture methods. Minimum inhibitory concentration and minimum bactericidal concentration of five antimicrobial agents (doxycycline, amoxicillin, tigecycline, metronidazole, and tinidazole against spirochetal forms of B. burgdorferi were evaluated using the standard published microdilution technique. The susceptibility of spirochetal and round body forms to the antibiotics was then tested using fluorescent microscopy (BacLight™ viability staining and dark field microscopy (direct cell counting, and these results were compared with the microdilution technique. Qualitative and quantitative effects of the antibiotics against biofilm-like colonies were assessed using fluorescent microscopy and dark field microscopy, respectively.Results: Doxycycline reduced spirochetal structures ~90% but increased the number of round body forms about twofold. Amoxicillin reduced spirochetal forms by ~85%–90% and round body

  9. Complement factor H-related proteins CFHR2 and CFHR5 represent novel ligands for the infection-associated CRASP proteins of Borrelia burgdorferi.

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    Corinna Siegel

    Full Text Available BACKGROUND: One virulence property of Borrelia burgdorferi is its resistance to innate immunity, in particular to complement-mediated killing. Serum-resistant B. burgdorferi express up to five distinct complement regulator-acquiring surface proteins (CRASP which interact with complement regulator factor H (CFH and factor H-like protein 1 (FHL1 or factor H-related protein 1 (CFHR1. In the present study we elucidate the role of the infection-associated CRASP-3 and CRASP-5 protein to serve as ligands for additional complement regulatory proteins as well as for complement resistance of B. burgdorferi. METHODOLOGY/PRINCIPAL FINDINGS: To elucidate whether CRASP-5 and CRASP-3 interact with various human proteins, both borrelial proteins were immobilized on magnetic beads. Following incubation with human serum, bound proteins were eluted and separated by Glycine-SDS-PAGE. In addition to CFH and CFHR1, complement regulators CFHR2 and CFHR5 were identified as novel ligands for both borrelial proteins by employing MALDI-TOF. To further assess the contributions of CRASP-3 and CRASP-5 to complement resistance, a serum-sensitive B. garinii strain G1 which lacks all CFH-binding proteins was used as a valuable model for functional analyses. Both CRASPs expressed on the B. garinii outer surface bound CFH as well as CFHR1 and CFHR2 in ELISA. In contrast, live B. garinii bound CFHR1, CFHR2, and CFHR5 and only miniscute amounts of CFH as demonstrated by serum adsorption assays and FACS analyses. Further functional analysis revealed that upon NHS incubation, CRASP-3 or CRASP-5 expressing borreliae were killed by complement. CONCLUSIONS/SIGNIFICANCE: In the absence of CFH and the presence of CFHR1, CFHR2 and CFHR5, assembly and integration of the membrane attack complex was not efficiently inhibited indicating that CFH in co-operation with CFHR1, CFHR2 and CFHR5 supports complement evasion of B. burgdorferi.

  10. Evidence that RpoS (sigmaS) in Borrelia burgdorferi is controlled directly by RpoN (sigma54/sigmaN).

    Science.gov (United States)

    Smith, Alexandra H; Blevins, Jon S; Bachlani, Gulnaz N; Yang, Xiaofeng F; Norgard, Michael V

    2007-03-01

    The alternative sigma factor (RpoN-RpoS) pathway controls the expression of key virulence factors in Borrelia burgdorferi. However, evidence to support whether RpoN controls rpoS directly or, perhaps, indirectly via a transactivator has been lacking. Herein we provide biochemical and genetic evidence that RpoN directly controls rpoS in B. burgdorferi. PMID:17158681

  11. Effectiveness of Stevia Rebaudiana Whole Leaf Extract Against the Various Morphological Forms of Borrelia Burgdorferi in Vitro.

    Science.gov (United States)

    Theophilus, P A S; Victoria, M J; Socarras, K M; Filush, K R; Gupta, K; Luecke, D F; Sapi, E

    2015-12-01

    Lyme disease is a tick-borne multisystemic disease caused by Borrelia burgdorferi. Administering antibiotics is the primary treatment for this disease; however, relapse often occurs when antibiotic treatment is discontinued. The reason for relapse remains unknown, but recent studies suggested the possibilities of the presence of antibiotic resistant Borrelia persister cells and biofilms. In this study, we evaluated the effectiveness of whole leaf Stevia extract against B. burgdorferi spirochetes, persisters, and biofilm forms in vitro. The susceptibility of the different forms was evaluated by various quantitative techniques in addition to different microscopy methods. The effectiveness of Stevia was compared to doxycycline, cefoperazone, daptomycin, and their combinations. Our results demonstrated that Stevia had significant effect in eliminating B. burgdorferi spirochetes and persisters. Subculture experiments with Stevia and antibiotics treated cells were established for 7 and 14 days yielding, no and 10% viable cells, respectively compared to the above-mentioned antibiotics and antibiotic combination. When Stevia and the three antibiotics were tested against attached biofilms, Stevia significantly reduced B. burgdorferi forms. Results from this study suggest that a natural product such as Stevia leaf extract could be considered as an effective agent against B. burgdorferi. PMID:26716015

  12. Effectiveness of Stevia Rebaudiana Whole Leaf Extract Against the Various Morphological Forms of Borrelia Burgdorferi in Vitro

    Science.gov (United States)

    Theophilus, P. A. S.; Victoria, M. J.; Socarras, K. M.; Filush, K. R.; Gupta, K.; Luecke, D. F.; Sapi, E.

    2015-01-01

    Lyme disease is a tick-borne multisystemic disease caused by Borrelia burgdorferi. Administering antibiotics is the primary treatment for this disease; however, relapse often occurs when antibiotic treatment is discontinued. The reason for relapse remains unknown, but recent studies suggested the possibilities of the presence of antibiotic resistant Borrelia persister cells and biofilms. In this study, we evaluated the effectiveness of whole leaf Stevia extract against B. burgdorferi spirochetes, persisters, and biofilm forms in vitro. The susceptibility of the different forms was evaluated by various quantitative techniques in addition to different microscopy methods. The effectiveness of Stevia was compared to doxycycline, cefoperazone, daptomycin, and their combinations. Our results demonstrated that Stevia had significant effect in eliminating B. burgdorferi spirochetes and persisters. Subculture experiments with Stevia and antibiotics treated cells were established for 7 and 14 days yielding, no and 10% viable cells, respectively compared to the above-mentioned antibiotics and antibiotic combination. When Stevia and the three antibiotics were tested against attached biofilms, Stevia significantly reduced B. burgdorferi forms. Results from this study suggest that a natural product such as Stevia leaf extract could be considered as an effective agent against B. burgdorferi. PMID:26716015

  13. Borrelia burgdorferi antibodies in dogs from Cotia county, São Paulo State, Brazil

    Directory of Open Access Journals (Sweden)

    JOPPERT Adriana Marques

    2001-01-01

    Full Text Available Dogs sera samples collected from Cotia County, São Paulo were tested using indirect immunoenzymatic test (ELISA in order to study Lyme disease serology in dogs. ELISA method was standardized and G39/40 North American strain of Borrelia burgdorferi was used as antigen. Positive results were confirmed employing the Western blotting technique. Because of the possibility of cross-reactions, sera were also tested for different serological strains of Leptospira interrogans and L. biflexa using microscopic sera agglutination test. Twenty-three of 237 (9.7% serum samples were positive in the ELISA; 20 of them (86.9% were confirmed by the Western blotting, what suggests that Cotia may be a risk area for Lyme disease. Although 4 samples (1.7% were positive for Lyme disease and leptospirosis, no correlation was found between the results (X² = 0.725; p = 0.394 what suggests absence of serological cross reactivity.

  14. Phenotypic and Genetic Characterization of a Novel Borrelia burgdorferi Sensu Lato Isolate from a Patient with Lyme Borreliosis

    OpenAIRE

    Wang, Guiqing; Dam, Alje P. van; Dankert, Jacob

    1999-01-01

    Borrelia burgdorferi sensu lato A14S was cultured from a skin biopsy specimen of a patient with erythema migrans in The Netherlands. This isolate had a unique DNA fingerprint pattern compared to 135 other B. burgdorferi sensu lato isolates. In this study, the isolate A14S was further characterized by protein analysis with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and reactivity with various monoclonal antibodies. In addition, the 16S rRNA, ospA, and ospC genes, as w...

  15. Role of Acetyl-Phosphate in Activation of the Rrp2-RpoN-RpoS Pathway in Borrelia burgdorferi

    OpenAIRE

    Haijun Xu; Caimano, Melissa J.; Tao Lin; Ming He; Radolf, Justin D.; Norris, Steven J; Frank Gherardini; Wolfe, Alan J.; X Frank Yang

    2010-01-01

    Borrelia burgdorferi, the Lyme disease spirochete, dramatically alters its transcriptome and proteome as it cycles between the arthropod vector and mammalian host. During this enzootic cycle, a novel regulatory network, the Rrp2-RpoN-RpoS pathway (also known as the σ(54)-σ(S) sigma factor cascade), plays a central role in modulating the differential expression of more than 10% of all B. burgdorferi genes, including the major virulence genes ospA and ospC. However, the mechanism(s) by which th...

  16. Detection of Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in hard ticks (Acari, Ixodidae) parasitizing bats in Poland.

    Science.gov (United States)

    Piksa, Krzysztof; Stańczak, Joanna; Biernat, Beata; Górz, Andrzej; Nowak-Chmura, Magdalena; Siuda, Krzysztof

    2016-04-01

    A total of 491 Ixodes vespertilionis and 8 Ixodes ricinus collected from bats and cave walls in southern Poland between 2010 and 2012 were examined by the polymerase chain reaction for tick-transmitted pathogens. PCR analysis for Borrelia burgdorferi s.l., Rickettsia spp., and Anaplasma phagocytophilum yielded negative results for all I. vespertilionis. DNA of Rickettsia helvetica was detected in three specimens of I. ricinus attached to Rhinolophus hipposideros or Myotis myotis, while Borrelia garinii was found in one tick parasitizing Myotis daubentonii. These pathogens were recorded for the first time in hard ticks that parasitized bats. PMID:26833325

  17. NMR structure of an acyl-carrier protein from Borrelia burgdorferi

    International Nuclear Information System (INIS)

    The high-resolution NMR structure of the acyl-carrier protein from the pathogen B. burgdorferi determined to a r.m.s. deviation of 0.4 Å over the protein backbone is reported. The NMR structure was determined using multidimensional NMR spectroscopy and consists of four α-helices and two 310-helices. Structural comparison reveals that this protein is highly similar to the acyl-carrier protein from A. aeolicus. Nearly complete resonance assignment and the high-resolution NMR structure of the acyl-carrier protein from Borrelia burgdorferi, a target of the Seattle Structural Genomics Center for Infectious Disease (SSGCID) structure-determination pipeline, are reported. This protein was chosen as a potential target for drug-discovery efforts because of its involvement in fatty-acid biosynthesis, an essential metabolic pathway, in bacteria. It was possible to assign >98% of backbone resonances and >92% of side-chain resonances using multidimensional NMR spectroscopy. The NMR structure was determined to a backbone r.m.s.d. of 0.4 Å and contained four α-helices and two 310-helices. A structure-homology search revealed that this protein is highly similar to the acyl-carrier protein from Aquifex aeolicus

  18. Cardiac Tropism of Borrelia burgdorferi: An Autopsy Study of Sudden Cardiac Death Associated with Lyme Carditis.

    Science.gov (United States)

    Muehlenbachs, Atis; Bollweg, Brigid C; Schulz, Thadeus J; Forrester, Joseph D; DeLeon Carnes, Marlene; Molins, Claudia; Ray, Gregory S; Cummings, Peter M; Ritter, Jana M; Blau, Dianna M; Andrew, Thomas A; Prial, Margaret; Ng, Dianna L; Prahlow, Joseph A; Sanders, Jeanine H; Shieh, Wun Ju; Paddock, Christopher D; Schriefer, Martin E; Mead, Paul; Zaki, Sherif R

    2016-05-01

    Fatal Lyme carditis caused by the spirochete Borrelia burgdorferi rarely is identified. Here, we describe the pathologic, immunohistochemical, and molecular findings of five case patients. These sudden cardiac deaths associated with Lyme carditis occurred from late summer to fall, ages ranged from young adult to late 40s, and four patients were men. Autopsy tissue samples were evaluated by light microscopy, Warthin-Starry stain, immunohistochemistry, and PCR for B. burgdorferi, and immunohistochemistry for complement components C4d and C9, CD3, CD79a, and decorin. Post-mortem blood was tested by serology. Interstitial lymphocytic pancarditis in a relatively characteristic road map distribution was present in all cases. Cardiomyocyte necrosis was minimal, T cells outnumbered B cells, plasma cells were prominent, and mild fibrosis was present. Spirochetes in the cardiac interstitium associated with collagen fibers and co-localized with decorin. Rare spirochetes were seen in the leptomeninges of two cases by immunohistochemistry. Spirochetes were not seen in other organs examined, and joint tissue was not available for evaluation. Although rare, sudden cardiac death caused by Lyme disease might be an under-recognized entity and is characterized by pancarditis and marked tropism of spirochetes for cardiac tissues. PMID:26968341

  19. Borrelia burgdorferi strain-specific Osp C-mediated immunity in mice.

    Science.gov (United States)

    Bockenstedt, L K; Hodzic, E; Feng, S; Bourrel, K W; de Silva, A; Montgomery, R R; Fikrig, E; Radolf, J D; Barthold, S W

    1997-11-01

    Antibodies to the outer surface proteins (Osps) A, B, and C of the spirochete Borrelia burgdorferi can prevent infection in animal models of Lyme borreliosis. We have previously demonstrated that immune serum from mice infected with B. burgdorferi N40 can also prevent challenge infection and induce disease regression in infected mice. The antigens targeted by protective and disease-modulating antibodies are presently unknown, but they do not include Osp A or Osp B. Because Osp C antibodies are present in immune mouse serum, we investigated the ability of hyperimmune serum to recombinant Osp C (N40) to protect mice against challenge infection with N40 spirochetes. In both active and passive immunization studies, Osp C (N40) antiserum failed to protect mice from challenge infection with cultured organisms. Mice actively immunized with recombinant Osp C (N40) were susceptible to tick-borne challenge infection, and nymphal ticks remained infected after feeding on Osp C-hyperimmunized mice. In contrast, similar immunization studies performed with Osp C (PKo) antiserum prevented challenge infection of mice with a clone of PKo spirochetes pathogenic for mice. Both Osp C (N40) and Osp C (PKo) antisera showed minimal in vitro borreliacidal activity, and immunofluorescence studies localized Osp C beneath the outer membrane of both N40 and PKo spirochetes. We conclude that Osp C antibody-mediated immunity is strain specific and propose that differences in Osp C surface expression by spirochetes in vivo may account for strain-specific immunity. PMID:9353047

  20. First report of Borrelia burgdorferi sensu lato in two threatened carnivores: the Marbled polecat, Vormela peregusna and the European mink, Mustela lutreola (Mammalia: Mustelidae

    Directory of Open Access Journals (Sweden)

    Gherman Călin M

    2012-08-01

    Full Text Available Abstract Background Lyme disease is a widespread cosmopolitan zoonosis caused by species belonging to the genus Borrelia. It is transmitted from animal reservoir hosts to humans through hard - ticks of genus Ixodes which are vectors of the disease. Case presentation Borrelia burgdorferi sensu lato infection was identified in a marbled polecat, Vormela peregusna, and two European minks, Mustela lutreola, from Romania, by PCR. RFLP revealed the presence of a single genospecies, Borrelia burgdorferi sensu stricto. Conclusions This is the first report of the Lyme disease spirochetes in the two mentioned hosts.

  1. Serum antibodies to whole-cell and recombinant antigens of Borrelia burgdorferi in cottontail rabbits.

    Science.gov (United States)

    Magnarelli, Louis A; Norris, Steven J; Fikrig, Erol

    2012-01-01

    Archived serum samples, from 95 eastern cottontail rabbits (Sylvilagus floridanus) captured in New York, New York, USA and Millbrook, New York, USA, during 1985-86, were analyzed in solid-phase enzyme-linked immunosorbent assays (ELISA) for total and class-specific immunoglobulin (Ig) M antibodies to whole-cell or recombinant antigens of Borrelia burgdorferi sensu stricto. Using a polyvalent conjugate, rabbit sera contained antibodies to whole-cell and recombinant antigens (protein [p]35, p37, or VlsE) during different seasons, but there was no reactivity to outer surface protein (Osp)A or OspB. Seventy-six of the 102 sera (75%) analyzed were reactive with one or more of the antigens; 61 of the positive samples (80%) reacted to whole-cell antigens, followed by results for the p35 (58%, 44/76), VlsE (43%, 33/76), and p37 (29%, 22/ 76) antigens. Fifty-eight sera (76%) contained antibodies to the VlsE or p35 antigens with or without reactivity to whole-cell antigens. High antibody titers (≥1:2,560) recorded for 52 sera indicate robust antibody production. In analyses for IgM antibodies in an ELISA containing whole-cell antigens, there were 30 positive sera; titers ranged from 1:160 to 1:640. There was minimal cross-reactivity when rabbit antisera to Treponema pallidum or four serovars of Leptospira interrogans were screened against B. burgdorferi antigens. Based on more-specific results, VlsE and p35 antigens appear to be useful markers for detecting possible B. burgdorferi infections. PMID:22247369

  2. Lyme Borreliosis: is there a preexisting (natural) variation in antimicrobial susceptibility among Borrelia burgdorferi strains?

    Science.gov (United States)

    Hodzic, Emir

    2015-01-01

    The development of antibiotics changed the world of medicine and has saved countless human and animal lives. Bacterial resistance/tolerance to antibiotics have spread silently across the world and has emerged as a major public health concern. The recent emergence of pan-resistant bacteria can overcome virtually any antibiotic and poses a major problem for their successful control. Selection for antibiotic resistance may take place where an antibiotic is present in the skin, gut, and other tissues of humans and animals and in the environment. Borrelia burgdorferi, the etiological agents of Lyme borreliosis, evades host immunity and establishes persistent infections in its mammalian hosts. The persistent infection poses a challenge to the effective antibiotic treatment, as demonstrated in various animal models. An increasingly heterogeneous subpopulation of replicatively attenuated spirochetes arises following treatment, and these persistent antimicrobial tolerant/resistant spirochetes are non-cultivable. The non-cultivable spirochetes resurge in multiple tissues at 12 months after treatment, with B. burgdorferi-specific DNA copy levels nearly equivalent to those found in shame-treated experimental animals. These attenuated spirochetes remain viable, but divide slowly, thereby being tolerant to antibiotics. Despite the continued non-cultivable state, RNA transcription of multiple B. burgdorferi genes was detected in host tissues, spirochetes were acquired by xenodiagnostic ticks, and spirochetal forms could be visualized within ticks and mouse tissues. A number of host cytokines were up- or down-regulated in tissues of both shame- and antibiotic-treated mice in the absence of histopathology, indicating a lack of host response to the presence of antimicrobial tolerant/resistant spirochetes. PMID:26295288

  3. Identification of Additional Anti-Persister Activity against Borrelia burgdorferi from an FDA Drug Library.

    Science.gov (United States)

    Feng, Jie; Weitner, Megan; Shi, Wanliang; Zhang, Shuo; Sullivan, David; Zhang, Ying

    2015-01-01

    Lyme disease is a leading vector-borne disease in the United States. Although the majority of Lyme patients can be cured with standard 2-4 week antibiotic treatment, 10%-20% of patients continue to suffer from prolonged post-treatment Lyme disease syndrome (PTLDS). While the cause for this is unclear, persisting organisms not killed by current Lyme antibiotics may be involved. In our previous study, we screened an FDA drug library and reported 27 top hits that showed high activity against Borrelia persisters. In this study, we present the results of an additional 113 active hits that have higher activity against the stationary phase B. burgdorferi than the currently used Lyme antibiotics. Many antimicrobial agents (antibiotics, antivirals, antifungals, anthelmintics or antiparasitics) used for treating other infections were found to have better activity than the current Lyme antibiotics. These include antibacterials such as rifamycins (3-formal-rifamycin, rifaximin, rifamycin SV), thiostrepton, quinolone drugs (sarafloxacin, clinafloxacin, tosufloxacin), and cell wall inhibitors carbenicillin, tazobactam, aztreonam; antifungal agents such as fluconazole, mepartricin, bifonazole, climbazole, oxiconazole, nystatin; antiviral agents zanamivir, nevirapine, tilorone; antimalarial agents artemisinin, methylene blue, and quidaldine blue; antihelmintic and antiparasitic agents toltrazuril, tartar emetic, potassium antimonyl tartrate trihydrate, oxantel, closantel, hycanthone, pyrimethamine, and tetramisole. Interestingly, drugs used for treating other non-infectious conditions including verteporfin, oltipraz, pyroglutamic acid, pidolic acid, and dextrorphan tartrate, that act on the glutathione/γ-glutamyl pathway involved in protection against free radical damage, and also the antidepressant drug indatraline, were found to have high activity against stationary phase B. burgdorferi. Among the active hits, agents that affect cell membranes, energy production, and reactive

  4. Identification of Additional Anti-Persister Activity against Borrelia burgdorferi from an FDA Drug Library

    Directory of Open Access Journals (Sweden)

    Jie Feng

    2015-09-01

    Full Text Available Lyme disease is a leading vector-borne disease in the United States. Although the majority of Lyme patients can be cured with standard 2–4 week antibiotic treatment, 10%–20% of patients continue to suffer from prolonged post-treatment Lyme disease syndrome (PTLDS. While the cause for this is unclear, persisting organisms not killed by current Lyme antibiotics may be involved. In our previous study, we screened an FDA drug library and reported 27 top hits that showed high activity against Borrelia persisters. In this study, we present the results of an additional 113 active hits that have higher activity against the stationary phase B. burgdorferi than the currently used Lyme antibiotics. Many antimicrobial agents (antibiotics, antivirals, antifungals, anthelmintics or antiparasitics used for treating other infections were found to have better activity than the current Lyme antibiotics. These include antibacterials such as rifamycins (3-formal-rifamycin, rifaximin, rifamycin SV, thiostrepton, quinolone drugs (sarafloxacin, clinafloxacin, tosufloxacin, and cell wall inhibitors carbenicillin, tazobactam, aztreonam; antifungal agents such as fluconazole, mepartricin, bifonazole, climbazole, oxiconazole, nystatin; antiviral agents zanamivir, nevirapine, tilorone; antimalarial agents artemisinin, methylene blue, and quidaldine blue; antihelmintic and antiparasitic agents toltrazuril, tartar emetic, potassium antimonyl tartrate trihydrate, oxantel, closantel, hycanthone, pyrimethamine, and tetramisole. Interestingly, drugs used for treating other non-infectious conditions including verteporfin, oltipraz, pyroglutamic acid, pidolic acid, and dextrorphan tartrate, that act on the glutathione/γ-glutamyl pathway involved in protection against free radical damage, and also the antidepressant drug indatraline, were found to have high activity against stationary phase B. burgdorferi. Among the active hits, agents that affect cell membranes, energy

  5. Clinical characteristics associated with Borrelia burgdorferi sensu lato skin culture results in patients with erythema migrans.

    Directory of Open Access Journals (Sweden)

    Franc Strle

    Full Text Available Clinical characteristics associated with isolation of Borrelia burgdorferi sensu lato from skin have not been fully evaluated. To gain insight into predictors for a positive EM skin culture, we compared basic demographic, epidemiologic, and clinical data in 608 culture-proven and 501 culture-negative adult patients with solitary EM. A positive Borrelia spp. skin culture was associated with older age, a time interval of >2 days between tick bite and onset of the skin lesion, EM ≥ 5 cm in diameter, and location of the lesion on the extremities, whereas several other characteristics used as clinical case definition criteria for the diagnosis of EM (such as tick bite at the site of later EM, information on expansion of the skin lesion, central clearing were not. A patient with a 15-cm EM lesion had almost 3-fold greater odds for a positive skin culture than patients with a 5-cm lesion. Patients with a free time interval between the tick bite and onset of EM had the same probability of a positive skin culture as those who did not recall a tick bite (OR=1.02; however, the two groups had >3-fold greater odds for EM positivity than patients who reported a tick bite with no interval between the bite and onset of the lesion. In conclusion, several yet not all clinical characteristics used in EM case definitions were associated with positive Borrelia spp. skin culture. The findings are limited to European patients with solitary EM caused predominantly by B. afzelii but may not be valid for other situations.

  6. Inhibition of the Production of Anti-OspA Borreliacidal Antibody with T Cells from Hamsters Vaccinated against Borrelia burgdorferi

    OpenAIRE

    Jensen, Jani R.; Du Chateau, Brian K.; Munson, Erik L.; Callister, Steven M.; Schell, Ronald F.

    1998-01-01

    The serious morbidity associated with Lyme borreliosis has focused considerable effort on the development of a comprehensive vaccine for protection against infection with Borrelia burgdorferi. Induction of borreliacidal antibody by vaccination or infection has been shown to correlate with protection of humans and animals against infection with the Lyme spirochete. In this report, we showed that high levels of borreliacidal antibody (titer of 1,280) were produced in vitro when T and B cells fr...

  7. Effect of salivary gland extract from Ixodes ricinus ticks on the proliferation of Borrelia burgdorferi sensu stricto in vivo

    Czech Academy of Sciences Publication Activity Database

    Macháčková, Martina; Oborník, Miroslav; Kopecký, Jan

    2006-01-01

    Roč. 53, č. 2 (2006), s. 153-158. ISSN 0015-5683 R&D Projects: GA ČR GA524/05/0811 Grant ostatní: Studentská grantová agentura, Biologická fakulta Jihočeské univerzity(CZ) SGA2004/009 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia burgdorferi * Ixodes ricinus * saliva -activated transmission Subject RIV: EC - Immunology Impact factor: 1.511, year: 2006

  8. Heterogeneity of outer membrane proteins in Borrelia burgdorferi: comparison of osp operons of three isolates of different geographic origins.

    OpenAIRE

    JONSSON, M; Noppa, L; Barbour, A G; Bergström, S

    1992-01-01

    Biochemical and immunochemical studies of the outer membrane proteins of Borrelia burgdorferi have shown that the OspA and OspB proteins from strains of different geographic origins may differ considerably in their reactivities with monoclonal antibodies and in their apparent molecular weights. To further characterize this variation in Osp proteins between strains, the osp operons and deduced translation products from two strains, one from Sweden (ACAI) and one from eastern Russia (Ip90), wer...

  9. Borrelia burgdorferi Infection-Associated Surface Proteins ErpP, ErpA, and ErpC Bind Human Plasminogen▿

    OpenAIRE

    Brissette, Catherine A.; Haupt, Katrin; Barthel, Diana; Cooley, Anne E.; Bowman, Amy; Skerka, Christina; Wallich, Reinhard; Zipfel, Peter F.; Kraiczy, Peter; Stevenson, Brian

    2008-01-01

    Host-derived plasmin plays a critical role in mammalian infection by Borrelia burgdorferi. The Lyme disease spirochete expresses several plasminogen-binding proteins. Bound plasminogen is converted to the serine protease plasmin and thereby may facilitate the bacterium's dissemination throughout the host by degrading extracellular matrix. In this work, we demonstrate plasminogen binding by three highly similar borrelial outer surface proteins, ErpP, ErpA, and ErpC, all of which are expressed ...

  10. PCR-Based Quantification of Borrelia burgdorferi Organisms in Canine Tissues over a 500-Day Postinfection Period

    OpenAIRE

    Straubinger, Reinhard K.

    2000-01-01

    Borrelia burgdorferi infection in beagle dogs was studied quantitatively with skin punch biopsy samples and blood samples collected at 4- and 2-week intervals, respectively, over a 500-day period. Thereafter, 25 tissue samples of each dog were collected for further analysis. Starting at day 120 after tick challenge, 12 dogs were treated with antibiotics (azithromycin, ceftriaxone, or doxycycline) for 30 consecutive days. Four dogs received no antibiotic therapy. Quantification of B. burgdorfe...

  11. Detection of Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in hard ticks (Acari, Ixodidae) parasitizing bats in Poland

    OpenAIRE

    Piksa, Krzysztof; Stańczak, Joanna; Biernat, Beata; Górz, Andrzej; Nowak-Chmura, Magdalena; Siuda, Krzysztof

    2016-01-01

    A total of 491 Ixodes vespertilionis and 8 Ixodes ricinus collected from bats and cave walls in southern Poland between 2010 and 2012 were examined by the polymerase chain reaction for tick-transmitted pathogens. PCR analysis for Borrelia burgdorferi s.l., Rickettsia spp., and Anaplasma phagocytophilum yielded negative results for all I. vespertilionis. DNA of Rickettsia helvetica was detected in three specimens of I. ricinus attached to Rhinolophus hipposideros or Myotis myotis, while Borrel...

  12. Borrelia burgdorferi sensu lato in Ixodes cf. neuquenensis and Ixodes sigelos ticks from the Patagonian region of Argentina.

    Science.gov (United States)

    Sebastian, Patrick S; Bottero, Maria Noelia Saracho; Carvalho, Luis; Mackenstedt, Ute; Lareschi, Marcela; Venzal, José M; Nava, Santiago

    2016-10-01

    This study was conducted to detect Borrelia burgdorferi sensu lato infection in ixodid ticks from the Patagonia region in the south of Argentina. Therefore, ticks were collected on rodents in the provinces of Chubut, Río Negro and Santa Cruz. These ticks were identified as nymphs of Ixodes cf. neuquenensis and Ixodes sigelos. The B. burgdorferi s.l. infection was tested by a battery of PCR methods targeting the gene flagellin (fla) and the rrfA-rrlB intergenic spacer region (IGS). Three pools of I. sigelos nymphs from Chubut and Santa Cruz provinces as well as one pool of I. cf. neuquenensis nymphs from Río Negro province were tested positive in the fla-PCR. The samples of I. sigelos were also positive for the IGS-PCR. Phylogenetically, the haplotypes found in the positive ticks belong to the B. burgdorferi s.l. complex, and they were closely related to Borrelia chilensis, a genospecies isolated from Ixodes stilesi in Chile. The pathogenic relevance of the Borrelia genospecies detected in both I. neuquenensis and I. sigelos is unknown. PMID:27372197

  13. Isolation of live Borrelia burgdorferi sensu lato spirochaetes from patients with undefined disorders and symptoms not typical for Lyme borreliosis.

    Science.gov (United States)

    Rudenko, N; Golovchenko, M; Vancova, M; Clark, K; Grubhoffer, L; Oliver, J H

    2016-03-01

    Lyme borreliosis is a multisystem disorder with a diverse spectrum of clinical manifestations, caused by spirochaetes of the Borrelia burgdorferi sensu lato complex. It is an infectious disease that can be successfully cured by antibiotic therapy in the early stages; however, the possibility of the appearance of persistent signs and symptoms of disease following antibiotic treatment is recognized. It is known that Lyme borreliosis mimics multiple diseases that were never proven to have a spirochaete aetiology. Using complete modified Kelly-Pettenkofer medium we succeeded in cultivating live B. burgdorferi sensu lato spirochaetes from samples taken from people who suffered from undefined disorders, had symptoms not typical for Lyme borreliosis, but who had undergone antibiotic treatment due to a suspicion of having Lyme disease even though they were seronegative. We report the first recovery of live B. burgdorferi sensu stricto from residents of southeastern USA and the first successful cultivation of live Borrelia bissettii-like strain from residents of North America. Our results support the fact that B. bissettii is responsible for human Lyme borreliosis worldwide along with B. burgdorferi s.s. The involvement of new spirochaete species in Lyme borreliosis changes the understanding and recognition of clinical manifestations of this disease. PMID:26673735

  14. CD14 cooperates with complement receptor 3 to mediate MyD88-independent phagocytosis of Borrelia burgdorferi

    Science.gov (United States)

    Hawley, Kelly L.; Olson, Chris M.; Iglesias-Pedraz, Juan M.; Navasa, Nicolás; Cervantes, Jorge L.; Caimano, Melissa J.; Izadi, Hooman; Ingalls, Robin R.; Pal, Utpal; Salazar, Juan C.; Radolf, Justin D.; Anguita, Juan

    2012-01-01

    Phagocytosis of Borrelia burgdorferi, the causative agent of Lyme disease, is a poorly understood process, despite its importance during the host immune response to infection. B. burgdorferi has been shown to bind to different receptors on the surface of phagocytic cells, including the β2 integrin, complement receptor 3 (CR3). However, whether these receptors mediate the phagocytosis of the spirochete remains unknown. We now demonstrate that CR3 mediates the phagocytosis of the spirochete by murine macrophages and human monocytes. Interaction of B. burgdorferi with the integrin is not sufficient, however, to internalize the spirochete; phagocytosis requires the interaction of CR3 with the GPI-anchored protein, CD14, independently of TLR/MyD88-induced or inside-out signals. Interestingly, the absence of CR3 leads to marked increases in the production of TNF in vitro and in vivo, despite reduced spirochetal uptake. Furthermore, the absence of CR3 during infection with B. burgdorferi results in the inefficient control of bacterial burdens in the heart and increased Lyme carditis. Overall, our data identify CR3 as a MyD88-independent phagocytic receptor for B. burgdorferi that also participates in the modulation of the proinflammatory output of macrophages. These data also establish a unique mechanism of CR3-mediated phagocytosis that requires the direct cooperation of GPI-anchored proteins. PMID:22232682

  15. The multifaceted responses of primary human astrocytes and brain microvascular endothelial cells to the Lyme disease spirochete, Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Catherine A. Brissette

    2013-08-01

    Full Text Available The vector-borne pathogen, Borrelia burgdorferi, causes a multi-system disorder including neurological complications. These neurological disorders, collectively termed neuroborreliosis, can occur in up to 15% of untreated patients. The neurological symptoms are probably a result of a glial-driven, host inflammatory response to the bacterium. However, the specific contributions of individual glial and other support cell types to the pathogenesis of neuroborreliosis are relatively unexplored. The goal of this project was to characterize specific astrocyte and endothelial cell responses to B. burgdorferi. Primary human astrocytes and primary HBMEC (human brain microvascular endothelial cells were incubated with B. burgdorferi over a 72-h period and the transcriptional responses to the bacterium were analyzed by real-time PCR arrays. There was a robust increase in several surveyed chemokine and related genes, including IL (interleukin-8, for both primary astrocytes and HBMEC. Array results were confirmed with individual sets of PCR primers. The production of specific chemokines by both astrocytes and HBMEC in response to B. burgdorferi, including IL-8, CXCL-1, and CXCL-10, were confirmed by ELISA. These results demonstrate that primary astrocytes and HBMEC respond to virulent B. burgdorferi by producing a number of chemokines. These data suggest that infiltrating phagocytic cells, particularly neutrophils, attracted by chemokines expressed at the BBB (blood–brain barrier may be important contributors to the early inflammatory events associated with neuroborreliosis.

  16. Diagnostic Value of Proteins of Three Borrelia Species (Borrelia burgdorferi Sensu Lato) and Implications for Development and Use of Recombinant Antigens for Serodiagnosis of Lyme Borreliosis in Europe

    OpenAIRE

    Hauser, Ulrike; Lehnert, Gisela; Wilske, Bettina

    1998-01-01

    More and more assays for the serodiagnosis of Lyme borreliosis (LB) are based on recombinant antigens. However, so far, there is no consensus as to which are the most specific and sensitive proteins and how they should be used in combination to obtain tests with the best discrimination abilities. The present study was preceded by a detailed analysis of Western blots (WB) using whole-cell lysates of Borrelia burgdorferi sensu stricto strain PKa2, B. afzelii PKo, and B. garinii PBi (U. Hauser, ...

  17. BB0324 and BB0028 are constituents of the Borrelia burgdorferi β-barrel assembly machine (BAM complex

    Directory of Open Access Journals (Sweden)

    Lenhart Tiffany R

    2012-04-01

    Full Text Available Abstract Background Similar to Gram-negative bacteria, the outer membrane (OM of the pathogenic spirochete, Borrelia burgdorferi, contains integral OM-spanning proteins (OMPs, as well as membrane-anchored lipoproteins. Although the mechanism of OMP biogenesis is still not well-understood, recent studies have indicated that a heterooligomeric OM protein complex, known as BAM (β-barrel assembly machine is required for proper assembly of OMPs into the bacterial OM. We previously identified and characterized the essential β-barrel OMP component of this complex in B. burgdorferi, which we determined to be a functional BamA ortholog. Results In the current study, we report on the identification of two additional protein components of the B. burgdorferi BAM complex, which were identified as putative lipoproteins encoded by ORFs BB0324 and BB0028. Biochemical assays with a BamA-depleted B. burgdorferi strain indicate that BB0324 and BB0028 do not readily interact with the BAM complex without the presence of BamA, suggesting that the individual B. burgdorferi BAM components may associate only when forming a functional BAM complex. Cellular localization assays indicate that BB0324 and BB0028 are OM-associated subsurface lipoproteins, and in silico analyses indicate that BB0324 is a putative BamD ortholog. Conclusions The combined data suggest that the BAM complex of B. burgdorferi contains unique protein constituents which differ from those found in other proteobacterial BAM complexes. The novel findings now allow for the B. burgdorferi BAM complex to be further studied as a model system to better our understanding of spirochetal OM biogenesis in general.

  18. PCR-Based quantification of Borrelia burgdorferi organisms in canine tissues over a 500-Day postinfection period.

    Science.gov (United States)

    Straubinger, R K

    2000-06-01

    Borrelia burgdorferi infection in beagle dogs was studied quantitatively with skin punch biopsy samples and blood samples collected at 4- and 2-week intervals, respectively, over a 500-day period. Thereafter, 25 tissue samples of each dog were collected for further analysis. Starting at day 120 after tick challenge, 12 dogs were treated with antibiotics (azithromycin, ceftriaxone, or doxycycline) for 30 consecutive days. Four dogs received no antibiotic therapy. Quantification of B. burgdorferi DNA was done with an ABI Prism 7700 Sequence Detection System with oligonucleotide primers and a fluorescence-labeled probe designed to specifically amplify a fragment of the ospA gene of B. burgdorferi strain N40. All 16 dogs became infected with B. burgdorferi after tick challenge. In skin biopsy samples, spirochete numbers peaked at day 60 postinfection (dogs, and decreased to almost undetectable levels during the following 6 months. The number of B. burgdorferi organisms detected in skin biopsy samples was inversely correlated with the antibody levels measured by enzyme-linked immunosorbent assay. Antibiotic treatment reduced the amount of detectable spirochete DNA in skin tissue by a factor of 1,000 or more. At the end of the experiment, B. burgdorferi DNA was detectable at low levels (10(2) to 10(4) organisms per 100 microgram of extracted DNA) in multiple tissue samples regardless of treatment. However, more tissue samples of untreated dogs than of antibiotic-treated dogs were positive, and tissue samples of untreated dogs also were positive by culture. Only 1.6% of 576 blood samples of all dogs were positive for B. burgdorferi by PCR. PMID:10834975

  19. Lyme disease caused by Borrelia burgdorferi with two homeologous 16S rRNA genes: a case report

    Directory of Open Access Journals (Sweden)

    Lee SH

    2016-04-01

    Full Text Available Sin Hang Lee,1,21Pathology Department, Milford Hospital, Milford, CT, USA; 2Milford Molecular Diagnostics, Milford, CT, USA Abstract: Lyme disease (LD, the most common tick-borne disease in North America, is believed to be caused exclusively by Borrelia burgdorferi sensu stricto and is usually diagnosed by clinical evaluation and serologic assays. As reported previously in a peer-reviewed article, a 13-year-old boy living in the Northeast of the USA was initially diagnosed with LD based on evaluation of his clinical presentations and on serologic test results. The patient was treated with a course of oral doxycycline for 28 days, and the symptoms resolved. A year later, the boy developed a series of unusual symptoms and did not attend school for 1 year. A LD specialist reviewed the case and found the serologic test band patterns nondiagnostic of LD. The boy was admitted to a psychiatric hospital. After discharge from the psychiatric hospital, a polymerase chain reaction test performed in a winter month when the boy was 16 years old showed a low density of B. burgdorferi sensu lato in the blood of the patient, confirmed by partial 16S rRNA (ribosomal RNA gene sequencing. Subsequent DNA sequencing analysis presented in this report demonstrated that the spirochete isolate was a novel strain of B. burgdorferi with two homeologous 16S rRNA genes, which has never been reported in the world literature. This case report shows that direct DNA sequencing is a valuable tool for reliable molecular diagnosis of Lyme and related borrelioses, as well as for studies of the diversity of the causative agents of LD because LD patients infected by a rare or novel borrelial variant may produce an antibody pattern that can be different from the pattern characteristic of an infection caused by a typical B. burgdorferi sensu stricto strain. Keywords: Lyme disease, Borrelia burgdorferi, homeologous 16S rRNA genes, DNA sequencing

  20. Dual role of Fcγ receptors in host defense and disease in Borrelia burgdorferi-infected mice

    Directory of Open Access Journals (Sweden)

    Alexia Anne Belperron

    2014-06-01

    Full Text Available Arthritis in mice infected with the Lyme disease spirochete, Borrelia burgdorferi, results from the influx of innate immune cells responding to the pathogen in the joint and is influenced in part by mouse genetics. Production of inflammatory cytokines by innate immune cells in vitro is largely mediated by Toll-like receptor (TLR interaction with Borrelia lipoproteins, yet surprisingly mice deficient in TLR2 or the TLR signaling molecule MyD88 still develop arthritis comparable to that seen in wild type mice after B. burgdorferi infection. These findings suggest that other, MyD88-independent inflammatory pathways can contribute to arthritis expression. Clearance of B. burgdorferi is dependent on the production of specific antibody and phagocytosis of the organism. As Fc receptors (FcγR are important for IgG-mediated clearance of immune complexes and opsonized particles by phagocytes, we examined the role that FcγR play in host defense and disease in B. burgdorferi-infected mice. B. burgdorferi-infected mice deficient in the Fc receptor common gamma chain (FcεRγ-/- mice harbored ~10 fold more spirochetes than similarly infected wild type mice, and this was associated with a transient increase in arthritis severity. While the elevated pathogen burdens seen in B. burgdorferi-infected MyD88-/- mice were not affected by concomitant deficiency in FcγR, arthritis was reduced in FcεRγ-/-MyD88-/- mice in comparison to wild type or single knockout mice. Gene expression analysis from infected joints demonstrated that absence of both MyD88 and FcγR lowers mRNA levels of proteins involved in inflammation, including Cxcl1 (KC, Xcr1 (Gpr5, IL-1beta, and C reactive protein. Taken together, our results demonstrate a role for FcγR-mediated immunity in limiting pathogen burden and arthritis in mice during the acute phase of B. burgdorferi infection, and further suggest that this pathway contributes to the arthritis that develops in B. burgdorferi

  1. Trans-Atlantic exchanges have shaped the population structure of the Lyme disease agent Borrelia burgdorferi sensu stricto.

    Science.gov (United States)

    Castillo-Ramírez, S; Fingerle, V; Jungnick, S; Straubinger, R K; Krebs, S; Blum, H; Meinel, D M; Hofmann, H; Guertler, P; Sing, A; Margos, G

    2016-01-01

    The origin and population structure of Borrelia burgdorferi sensu stricto (s.s.), the agent of Lyme disease, remain obscure. This tick-transmitted bacterial species occurs in both North America and Europe. We sequenced 17 European isolates (representing the most frequently found sequence types in Europe) and compared these with 17 North American strains. We show that trans-Atlantic exchanges have occurred in the evolutionary history of this species and that a European origin of B. burgdorferi s.s. is marginally more likely than a USA origin. The data further suggest that some European human patients may have acquired their infection in North America. We found three distinct genetically differentiated groups: i) the outgroup species Borrelia bissettii, ii) two divergent strains from Europe, and iii) a group composed of strains from both the USA and Europe. Phylogenetic analysis indicated that different genotypes were likely to have been introduced several times into the same area. Our results demonstrate that irrespective of whether B. burgdorferi s.s. originated in Europe or the USA, later trans-Atlantic exchange(s) have occurred and have shaped the population structure of this genospecies. This study clearly shows the utility of next generation sequencing to obtain a better understanding of the phylogeography of this bacterial species. PMID:26955886

  2. Evaluation of Borrelia burgdorferi BbHtrA Protease as a Vaccine Candidate for Lyme Borreliosis in Mice.

    Directory of Open Access Journals (Sweden)

    Amy J Ullmann

    Full Text Available Borrelia burgdorferi synthesizes an HtrA protease (BbHtrA which is a surface-exposed, conserved protein within Lyme disease spirochetes with activity toward CheX and BmpD of Borrelia spp, as well as aggrecan, fibronectin and proteoglycans found in skin, joints and neural tissues of vertebrates. An antibody response against BbHtrA is observed in Lyme disease patients and in experimentally infected laboratory mice and rabbits. Given the surface location of BbHtrA on B. burgdorferi and its ability to elicit an antibody response in infected hosts, we explored recombinant BbHtrA as a potential vaccine candidate in a mouse model of tick-transmitted Lyme disease. We immunized mice with two forms of BbHtrA: the proteolytically active native form and BbHtrA ablated of activity by a serine to alanine mutation at amino acid 226 (BbHtrA(S226A. Although inoculation with either BbHtrA or BbHtrA(S226A produced high-titer antibody responses in C3H/HeJ mice, neither antigen was successful in protecting mice from B. burgdorferi challenge. These results indicate that the search for novel vaccine candidates against Lyme borreliosis remains a challenge.

  3. Reservoir competence of native North American birds for the Lyme disease spirochete, Borrelia burgdorferi

    Science.gov (United States)

    Ginsberg, H.S.; Buckley, P.A.; Balmforth, M.G.; Zhioua, E.; Mitra, Siddhartha; Buckley, F.G.

    2005-01-01

    Reservoir competence of the Lyme disease spirochete, Borrelia burgdorferi, was tested for six species of native North American birds: American Robin, Gray Catbird, Brown Thrasher, Eastern Towhee, Song Sparrow, and Northern Cardinal. Wild birds collected by mistnetting on Fire Island, NY, were held in a field lab in cages over water, and locally collected larval ticks were placed on the birds, harvested from the water after engorgement, and tested for infection by DFA after molting to the nymphal stage. American Robins were competent reservoirs, infecting 16.1% of larvae applied to wild-caught birds, compared to 0% of control ticks placed on uninfected lab mice. Robins that were previously infected in the lab by nymphal feeding infected 81.8% of applied larvae. Wild-caught Song Sparrows infected 4.8% of applied larvae, and 21.1% when infected by nymphal feeding. Results suggest moderate levels of reservoir competence for Northern Cardinals, lower levels for Gray Catbirds, and little evidence of reservoir competence for Eastern Towhees or Brown Thrashers. Lower infection rates in larvae applied to wild-caught birds compared to birds infected in the lab suggest that infected birds display temporal variability in infectiousness to larval ticks. Engorged larvae drop from birds abundantly during daylight hours, so the abundance of these bird species in the peridomestic environment suggests that they might contribute infected ticks to lawns and gardens.

  4. An inverted repeat in the ospC operator is required for induction in Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Dan Drecktrah

    Full Text Available Borrelia burgdorferi, the spirochete that causes Lyme disease, differentially regulates synthesis of the outer membrane lipoprotein OspC to infect its host. OspC is required to establish infection but then repressed in the mammal to avoid clearance by the adaptive immune response. Inverted repeats (IR upstream of the promoter have been implicated as an operator to regulate ospC expression. We molecularly dissected the distal inverted repeat (dIR of the ospC operator by site-directed mutagenesis at its endogenous location on the circular plasmid cp26. We found that disrupting the dIR but maintaining the proximal IR prevented induction of OspC synthesis by DNA supercoiling, temperature, and pH. Moreover, the base-pairing potential of the two halves of the dIR was more important than the nucleotide sequence in controlling OspC levels. These results describe a cis-acting element essential for the expression of the virulence factor OspC.

  5. Introduced Siberian chipmunks (Tamias sibiricus barberi) harbor more-diverse Borrelia burgdorferi sensu lato genospecies than native bank voles (Myodes glareolus)

    OpenAIRE

    Marsot, Maud; Sigaud, M.; Chapuis, J L; Ferquel, E.; Vourc'H, Gwenaël

    2011-01-01

    Little attention has been given in scientific literature to how introduced species may act as a new host for native infectious agents and modify the epidemiology of a disease. In this study, we investigated whether an introduced species, the Siberian chipmunk (Tamias sibiricus barberi), was a potentially new reservoir host for Borrelia burgdorferi sensu lato, the causative agent of Lyme disease. First, we ascertained whether chipmunks were infected by all of the B. burgdorferi sensu lato geno...

  6. Introduced Siberian Chipmunks (Tamias sibiricus barberi) Harbor More-Diverse Borrelia burgdorferi Sensu Lato Genospecies than Native Bank Voles (Myodes glareolus)▿

    OpenAIRE

    Marsot, M.; Sigaud, M.; Chapuis, J L; Ferquel, E.; Cornet, M.; Vourc'h, G

    2011-01-01

    Little attention has been given in scientific literature to how introduced species may act as a new host for native infectious agents and modify the epidemiology of a disease. In this study, we investigated whether an introduced species, the Siberian chipmunk (Tamias sibiricus barberi), was a potentially new reservoir host for Borrelia burgdorferi sensu lato, the causative agent of Lyme disease. First, we ascertained whether chipmunks were infected by all of the B. burgdorferi sensu lato geno...

  7. Automated purification of Borrelia burgdorferi s.l. PCR products with KingFisher{sup TM} magnetic particle processor prior to genome sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Maekinen, Johanna E-mail: johanna.makinen@utu.fi; Marttila, Harri; Viljanen, Matti K

    2001-07-01

    Borrelia burgdorferi sensu lato genospecies were differentiated by PCR-based sequencing of the borrelial flagellin gene. To evaluate the usefulness of KingFisher{sup TM} magnetic particle processor in PCR product purification, borrelia PCR products were purified with KingFisher{sup TM} magnetic particle processor prior to cycle sequencing and the quality of the sequence data received was analyzed. KingFisher was found to offer a rapid and reliable alternative for borrelial PCR product purification.

  8. Automated purification of Borrelia burgdorferi s.l. PCR products with KingFisher™ magnetic particle processor prior to genome sequencing

    Science.gov (United States)

    Mäkinen, Johanna; Marttila, Harri; Viljanen, Matti K.

    2001-01-01

    Borrelia burgdorferi sensu lato genospecies were differentiated by PCR-based sequencing of the borrelial flagellin gene. To evaluate the usefulness of KingFisher™ magnetic particle processor in PCR product purification, borrelia PCR products were purified with KingFisher™ magnetic particle processor prior to cycle sequencing and the quality of the sequence data received was analyzed. KingFisher was found to offer a rapid and reliable alternative for borrelial PCR product purification.

  9. Detection of four species of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected from roe deer (Capreolus capreolus) in The Netherlands.

    OpenAIRE

    Rijpkema, S G; Herbes, R. G.; Verbeek-De Kruif, N.; Schellekens, J. F.

    1996-01-01

    Roe deer (Capreolus capreolus) were investigated for their value as sentinel animals for Lyme borreliosis in the Netherlands. Serum was obtained from 114 roe deer, and 513 Ixodes ricinus, predominantly females (72%), were obtained from 47 animals (41%). The polymerase chain reaction was used to detect DNA of Borrelia burgdorferi sensu lato in a total of 190 ticks, comprising 106 engorged ticks and 84 non-engorged ticks. Borrelia DNA was detected in 24 engorged ticks (23%) and 26 non-engorged ...

  10. Automated purification of Borrelia burgdorferi s.l. PCR products with KingFisherTM magnetic particle processor prior to genome sequencing

    International Nuclear Information System (INIS)

    Borrelia burgdorferi sensu lato genospecies were differentiated by PCR-based sequencing of the borrelial flagellin gene. To evaluate the usefulness of KingFisherTM magnetic particle processor in PCR product purification, borrelia PCR products were purified with KingFisherTM magnetic particle processor prior to cycle sequencing and the quality of the sequence data received was analyzed. KingFisher was found to offer a rapid and reliable alternative for borrelial PCR product purification

  11. Polysynovitis in a horse due to [i]Borrelia burgdorferi[/i] sensu lato infection – Case study

    Directory of Open Access Journals (Sweden)

    Fabrizio Passamonti

    2015-05-01

    Full Text Available Lyme borreliosis (LB is a multi-systemic tick-borne disease affecting both humans and animals, including horses, and is caused by a group of interrelated spirochetes classified within the[i] Borrelia burgdorferi [/i]sensu lato (s.l. complex. Despite the high reported seroprevalence in the European equine population for [i]B. burgdorferi[/i] s.l., to-date no documented clinical cases have been described. A 6-year-old Paint gelding was referred with a history of three weeks of fever, intermittent lameness and digital flexor tendon sheath effusion of the right hind limb. Based on a strict diagnostic protocol, which included serological tests for infectious diseases and molecular investigations, a final diagnosis was made of polysynovitis due to [i]B. burgdorferi [/i]s.l. infection. An unreported aspect observed in this case was the absence of the pathogen DNA in two of the affected joints. To the authors’ knowledge, the case described represents the first documented clinical case of equine LB in Italy. Moreover, the absence of pathogen DNA in two of the affected joints observed in this case revealed a possible similarity with the same condition described in humans, where an immunomediated pathogenesis for arthropathy due to [i]B. burgdorferi[/i] s.l. infection is suspected. Since humans and horses share the same habitat, this report supports the role of the horse as potential sentinel for human biological risk.

  12. A Seventeen-Year Epidemiological Surveillance Study of Borrelia burgdorferi Infections in Two Provinces of Northern Spain

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    Lourdes Lledó

    2014-01-01

    Full Text Available This paper reports a 17-year seroepidemiological surveillance study of Borrelia burgdorferi infection, performed with the aim of improving our knowledge of the epidemiology of this pathogen. Serum samples (1,179 from patients (623, stratified with respect to age, sex, season, area of residence and occupation bitten by ticks in two regions of northern Spain were IFA-tested for B. burgdorferi antibodies. Positive results were confirmed by western blotting. Antibodies specific for B. burgdorferi were found in 13.3% of the patients; 7.8% were IgM positive, 9.6% were IgG positive, and 4.33% were both IgM and IgG positive. Five species of ticks were identified in the seropositive patients: Dermacentor marginatus (41.17% of such patients Dermacentor reticulatus (11.76%, Rhiphicephalus sanguineus (17.64%, Rhiphicephalus turanicus (5.88% and Ixodes ricinus (23.52%. B. burgdorferi DNA was sought by PCR in ticks when available. One tick, a D. reticulatus male, was found carrying the pathogen. The seroprevalence found was similar to the previously demonstrated in similar studies in Spain and other European countries.

  13. Genetic Variation of the Borrelia burgdorferi Gene vlsE Involves Cassette-Specific, Segmental Gene Conversion

    OpenAIRE

    Zhang, Jing-Ren; Norris, Steven J

    1998-01-01

    The Lyme disease spirochete Borrelia burgdorferi possesses 15 silent vls cassettes and a vls expression site (vlsE) encoding a surface-exposed lipoprotein. Segments of the silent vls cassettes have been shown to recombine with the vlsE cassette region in the mammalian host, resulting in combinatorial antigenic variation. Despite promiscuous recombination within the vlsE cassette region, the 5′ and 3′ coding sequences of vlsE that flank the cassette region are not subject to sequence variation...

  14. Diagnostic Value of PCR for Detection of Borrelia burgdorferi in Skin Biopsy and Urine Samples from Patients with Skin Borreliosis

    OpenAIRE

    Brettschneider, S; Bruckbauer, H.; Klugbauer, N; Hofmann, H.

    1998-01-01

    Skin biopsies of 36 patients with erythema migrans and acrodermatitis chronica atrophicans (ACA) before therapy and those of 8 patients after therapy were examined for Borrelia burgdorferi DNA by PCR. Skin biopsies of 27 patients with dermatological diseases other than Lyme borreliosis and those of 10 healthy persons were examined as controls. Two different primer sets targeting 23S rRNA (PCR I) and 66-kDa protein (PCR II) genes were used. PCR was performed with freshly frozen tissue (FFT) an...

  15. CsrA (BB0184) Is Not Involved in Activation of the RpoN-RpoS Regulatory Pathway in Borrelia burgdorferi

    OpenAIRE

    Ouyang, Zhiming; Zhou, Jianli; Norgard, Michael V.

    2014-01-01

    Borrelia burgdorferi encodes a homologue of the bacterial carbon storage regulator A (CsrA). Recently, it was reported that CsrA contributes to B. burgdorferi infectivity and is required for the activation of the central RpoN-RpoS regulatory pathway. However, many questions concerning the function of CsrA in B. burgdorferi gene regulation remain unanswered. In particular, there are conflicting reports concerning the molecular details of how CsrA may modulate rpoS expression and, thus, how Csr...

  16. Investigation of Borrelia burgdorferi genotypes in Australia obtained from erythema migrans tissue

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    Mayne PJ

    2012-07-01

    Full Text Available Peter J Mayne International Lyme and Associated Diseases Society, Bethesda, MD, USA The author is a member of the International Lyme and Associated Diseases Society (ILADSBackground: Lyme disease (LD is an emerging infectious disease in Australia. There has been controversy regarding endemic lyme disease in the country for over 20 years. Borrelia burgdorferi sensu stricto (Bbss and sensu lato (Bbsl are closely related spirochetal species that are the causative agents of LD in humans. Clinical transmission of this tick-borne disease is marked by a characteristic rash known as erythema migrans (EM. This study employed molecular techniques to demonstrate the spirochetal agent of Lyme disease isolated from EM biopsies of patients in Australia and then investigate their genetic diversity.Methods: Four patients who presented to the author's practice over a one-year period from mid 2010 to mid 2011 returned positive results on central tissue biopsy of EM lesions using polymerase chain reaction (PCR analysis. The findings were confirmed by DNA sequencing, and basic local alignment search tool (BLAST analysis was then used to genetically characterize the causative organisms.Results: Three isolates were identified as Bbss that lay genotypically between strains B31 and ZS7 and were then characterized as strain 64b. One of the three isolates though may have similarity to B. bissettii a Bbsl. The fourth isolate was more appropriately placed in the sensu lato group and appeared to be similar, but not identical to, a B. valaisiana-type isolate. In this study, a central biopsy taken within 6 days of infection was used instead of conventional sampling at the leading edge, and the merits of this are discussed.Conclusion: These patients acquired infection in Australia, further proving endemic LD on the continent. Central biopsy site of EM is a useful tool for PCR evaluation. BLAST searches suggest a genetic diversity of B. burgdorferi, which has implications

  17. Antibody profile to Borrelia burgdorferi in veterinarians from Nuevo León, Mexico, a non-endemic area of this zoonosis

    Science.gov (United States)

    Skinner-Taylor, Cassandra M.; Salinas, José A.; Arevalo-Niño, Katiushka; Galán-Wong, Luis J.; Maldonado, Guadalupe; Garza-Elizondo, Mario A.

    2016-01-01

    Objectives Lyme disease is a tick-borne disease caused by infections with Borrelia. Persons infected with Borrelia can be asymptomatic or can develop disseminated disease. Diagnosis and recognition of groups at risk of infection with Borrelia burgdorferi is of great interest to contemporary rheumatology. There are a few reports about Borrelia infection in Mexico, including lymphocytoma cases positive to B. burgdorferi sensu stricto by PCR and a patient with acrodermatitis chronica atrophicans. Veterinarians have an occupational risk due to high rates of tick contact. The aim of this work was to investigate antibodies to Borrelia in students at the Faculty of Veterinary Medicine and Zootechnics, at Nuevo León, Mexico, and determine the antibody profile to B. burgdorferi antigens. Material and methods Sera were screened using a C6 ELISA, IgG and IgM ELISA using recombinant proteins from B. burgdorferi, B. garinii and B. afzelii. Sera with positive or grey-zone values were tested by IgG Western blot to B. burgdorferi sensu stricto. Results All volunteers reported tick exposures and 72.5% remembered tick bites. Only nine persons described mild Lyme disease related symptoms, including headaches, paresthesias, myalgias and arthralgias. None of the volunteers reported erythema migrans. Nine samples were confirmed by IgG Western blot. The profile showed 89% reactivity to OspA, 67% to p83, and 45% to BmpA. Conclusions Positive sera samples shared antibody reactivity to the markers of late immune response p83 and BmpA, even if individuals did not present symptoms of Lyme arthritis or post-Lyme disease. The best criterion to diagnose Lyme disease in our country remains to be established, because it is probable that different strains coexist in Mexico. This is the first report of antibodies to B. burgdorferi in Latin American veterinarians. Veterinarians and high-risk people should be alert to take precautionary measures to prevent tick-borne diseases.

  18. An invasive mammal (the gray squirrel, Sciurus carolinensis) commonly hosts diverse and atypical genotypes of the zoonotic pathogen Borrelia burgdorferi sensu lato

    DEFF Research Database (Denmark)

    Millins, Caroline; Magierecka, Agnieszka; Gilbert, Lucy;

    2015-01-01

    Invasive vertebrate species can act as hosts for endemic pathogens and may alter pathogen community composition and dynamics. For the zoonotic pathogen Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, recent work shows invasive rodent species can be of high epidemiological importance...... and may support host specific strains. This study examined the role of grey squirrels (Sciurus carolinensis), (n=679), an invasive species in the United Kingdom (UK), as B. burgdorferi s.l. hosts. We found that grey squirrels were frequently infested with Ixodes ricinus, the main vector of B. burgdorferi...... in squirrels infested with ticks. Invasive grey squirrels appear to become infected with locally circulating strains of B. burgdorferi s.l., further studies are required to determine their role in community disease dynamics. Our findings highlight that the role of introduced host species in B. burgdorferi s...

  19. MicroRNA-146a provides feedback regulation of lyme arthritis but not carditis during infection with Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Robert B Lochhead

    2014-06-01

    Full Text Available MicroRNAs have been shown to be important regulators of inflammatory and immune responses and are implicated in several immune disorders including systemic lupus erythematosus and rheumatoid arthritis, but their role in Lyme borreliosis remains unknown. We performed a microarray screen for expression of miRNAs in joint tissue from three mouse strains infected with Borrelia burgdorferi. This screen identified upregulation of miR-146a, a key negative regulator of NF-κB signaling, in all three strains, suggesting it plays an important role in the in vivo response to B. burgdorferi. Infection of B6 miR-146a-/- mice with B. burgdorferi revealed a critical nonredundant role of miR-146a in modulating Lyme arthritis without compromising host immune response or heart inflammation. The impact of miR-146a was specifically localized to the joint, and did not impact lesion development or inflammation in the heart. Furthermore, B6 miR-146a-/- mice had elevated levels of NF-κB-regulated products in joint tissue and serum late in infection. Flow cytometry analysis of various lineages isolated from infected joint tissue of mice showed that myeloid cell infiltration was significantly greater in B6 miR-146a-/- mice, compared to B6, during B. burgdorferi infection. Using bone marrow-derived macrophages, we found that TRAF6, a known target of miR-146a involved in NF-κB activation, was dysregulated in resting and B. burgdorferi-stimulated B6 miR-146a-/- macrophages, and corresponded to elevated IL-1β, IL-6 and CXCL1 production. This dysregulated protein production was also observed in macrophages treated with IL-10 prior to B. burgdorferi stimulation. Peritoneal macrophages from B6 miR-146a-/- mice also showed enhanced phagocytosis of B. burgdorferi. Together, these data show that miR-146a-mediated regulation of TRAF6 and NF-κB, and downstream targets such as IL-1β, IL-6 and CXCL1, are critical for modulation of Lyme arthritis during chronic infection with B

  20. Cross-Species Surface Display of Functional Spirochetal Lipoproteins by Recombinant Borrelia burgdorferi

    OpenAIRE

    Zückert, Wolfram R.; Lloyd, Jill E.; Stewart, Philip E.; Rosa, Patricia A.; Barbour, Alan G.

    2004-01-01

    Surface-exposed lipoproteins of relapsing fever (RF) and Lyme borreliosis Borrelia spirochetes mediate certain interactions of the bacteria with their arthropod and vertebrate hosts. RF spirochetes such as Borrelia hermsii serially evade the host's antibody response by multiphasic antigenic variation of Vsp and Vlp proteins. Furthermore, the expression of Vsp1 and Vsp2 by Borrelia turicatae is associated with neurotropism and higher blood densities, respectively. In contrast to RF Borrelia sp...

  1. Prevalence of the Lyme Disease Spirochete, Borrelia burgdorferi, in Blacklegged Ticks, Ixodes scapularis at Hamilton-Wentworth, Ontario.

    Science.gov (United States)

    Scott, John D; Anderson, John F; Durden, Lance A; Smith, Morgan L; Manord, Jodi M; Clark, Kerry L

    2016-01-01

    Lyme disease has emerged as a major health concern in Canada, where the etiological agent, Borrelia burgdorferi sensu lato (s.l.), a spirochetal bacterium, is typically spread by the bite of certain ticks. This study explores the presence of B. burgdorferi s.l. in blacklegged ticks, Ixodes scapularis, collected at Dundas, Ontario (a locality within the region of Hamilton-Wentworth). Using passive surveillance, veterinarians and pet groomers were asked to collect blacklegged ticks from dogs and cats with no history of travel. Additionally, I. scapularis specimens were submitted from local residents and collected by flagging. Overall, 12 (41%) of 29 blacklegged ticks were infected with B. burgdorferi s.l. Using polymerase chain reaction (PCR) and DNA sequencing, two borrelial amplicons were characterized as B. burgdorferi sensu stricto (s.s.), a genospecies pathogenic to humans and certain domestic animals. Notably, three different vertebrate hosts each had two engorged I. scapularis females removed on the same day and, likewise, one cat had three repeat occurrences of this tick species. These multiple infestations suggest that a population of I. scapularis may be established in this area. The local public health unit has been underreporting the presence of B. burgdorferi s.l.-infected I. scapularis in the area encompassing Dundas. Our findings raise concerns about the need to erect tick warning signs in parkland areas. Veterinarians, medical professionals, public health officials, and the general public must be vigilant that Lyme disease-carrying blacklegged ticks pose a public health risk in the Dundas area and the surrounding Hamilton-Wentworth region. PMID:27226771

  2. Characterization of the RelBbu Regulon in Borrelia burgdorferi Reveals Modulation of Glycerol Metabolism by (pppGpp.

    Directory of Open Access Journals (Sweden)

    Julia V Bugrysheva

    Full Text Available The bacterial stringent response is triggered by deficiencies of available nutrients and other environmental stresses. It is mediated by 5'-triphosphate-guanosine-3'-diphosphate and 5'-diphosphate-guanosine-3'-diphosphate (collectively (pppGpp and generates global changes in gene expression and metabolism that enable bacteria to adapt to and survive these challenges. Borrelia burgdorferi encounters multiple stressors in its cycling between ticks and mammals that could trigger the stringent response. We have previously shown that the B. burgdorferi stringent response is mediated by a single enzyme, RelBbu, with both (pppGpp synthase and hydrolase activities, and that a B. burgdorferi 297 relBbu null deletion mutant was defective in adapting to stationary phase, incapable of down-regulating synthesis of rRNA and could not infect mice. We have now used this deletion mutant and microarray analysis to identify genes comprising the rel regulon in B. burgdorferi cultured at 34°C, and found that transcription of genes involved in glycerol metabolism is induced by relBbu. Culture of the wild type parental strain, the relBbu deletion mutant and its complemented derivative at 34°C and 25°C in media containing glucose or glycerol as principal carbon sources revealed a growth defect in the mutant, most evident at the lower temperature. Transcriptional analysis of the glp operon for glycerol uptake and metabolism in these three strains confirmed that relBbu was necessary and sufficient to increase transcription of this operon in the presence of glycerol at both temperatures. These results confirm and extend previous findings regarding the stringent response in B. burgdorferi. They also demonstrate that the stringent response regulates glycerol metabolism in this organism and is likely crucial for its optimal growth in ticks.

  3. Prevalence of the Lyme Disease Spirochete, Borrelia burgdorferi, in Blacklegged Ticks, Ixodes scapularis at Hamilton-Wentworth, Ontario

    Science.gov (United States)

    Scott, John D.; Anderson, John F.; Durden, Lance A.; Smith, Morgan L.; Manord, Jodi M.; Clark, Kerry L.

    2016-01-01

    Lyme disease has emerged as a major health concern in Canada, where the etiological agent, Borrelia burgdorferi sensu lato (s.l.), a spirochetal bacterium, is typically spread by the bite of certain ticks. This study explores the presence of B. burgdorferi s.l. in blacklegged ticks, Ixodes scapularis, collected at Dundas, Ontario (a locality within the region of Hamilton-Wentworth). Using passive surveillance, veterinarians and pet groomers were asked to collect blacklegged ticks from dogs and cats with no history of travel. Additionally, I. scapularis specimens were submitted from local residents and collected by flagging. Overall, 12 (41%) of 29 blacklegged ticks were infected with B. burgdorferi s.l. Using polymerase chain reaction (PCR) and DNA sequencing, two borrelial amplicons were characterized as B. burgdorferi sensu stricto (s.s.), a genospecies pathogenic to humans and certain domestic animals. Notably, three different vertebrate hosts each had two engorged I. scapularis females removed on the same day and, likewise, one cat had three repeat occurrences of this tick species. These multiple infestations suggest that a population of I. scapularis may be established in this area. The local public health unit has been underreporting the presence of B. burgdorferi s.l.-infected I. scapularis in the area encompassing Dundas. Our findings raise concerns about the need to erect tick warning signs in parkland areas. Veterinarians, medical professionals, public health officials, and the general public must be vigilant that Lyme disease-carrying blacklegged ticks pose a public health risk in the Dundas area and the surrounding Hamilton-Wentworth region. PMID:27226771

  4. Serological survey for antibodies to Borrelia burgdorferi in rodents and detection of spirochaetes in ticks and fleas in South Moravia (Czech Republic)

    Czech Academy of Sciences Publication Activity Database

    Juřicová, Zina; Halouzka, Jiří; Hubálek, Zdeněk

    2002-01-01

    Roč. 57, č. 3 (2002), s. 383-387. ISSN 0006-3088 R&D Projects: GA AV ČR IAA6087601; GA ČR GA206/00/1204 Institutional research plan: CEZ:AV0Z6093917 Keywords : Borrelia burgdorferi * Lyme disease Subject RIV: EE - Microbiology, Virology Impact factor: 0.169, year: 2002

  5. Presence of host-seeking Ixodes ricinus and their infection with Borrelia burgdorferi sensu lato in the Northern Apennines, Italy.

    Science.gov (United States)

    Ragagli, Charlotte; Mannelli, Alessandro; Ambrogi, Cecilia; Bisanzio, Donal; Ceballos, Leonardo A; Grego, Elena; Martello, Elisa; Selmi, Marco; Tomassone, Laura

    2016-06-01

    Host-seeking ticks were collected in the Northern Apennines, Italy, by dragging at 35 sites, at altitudes ranging from 680 and 1670 m above sea level (asl), from April to November, in 2010 and 2011. Ixodes ricinus (4431 larvae, 597 nymphs and 12 adults) and Haemaphysalis punctata (11,209 larvae, 313 nymphs, and 25 adults) were the most abundant species, followed by Haemaphysalis sulcata (20 larvae, five nymphs, and 13 adults), Dermacentor marginatus (42 larvae and two adults) and Ixodes hexagonus (one nymph). Greatest numbers of ticks were collected at locations characterised by southern exposure and limestone substratum, at altitudes Borrelia burgdorferi sensu lato (sl) in 294 host-seeking I. ricinus nymphs was 8.5 %. Borrelia garinii was the most frequently identified genospecies (64.0 % of positive nymphs), followed by B. valaisiana, B. burgdorferi sensu stricto, B. afzelii, and B. lusitaniae. Based upon the comparison with the results of previous studies at the same location, these research findings suggest the recent invasion of the study area by the tick vector and the agents of Lyme borreliosis. PMID:26964552

  6. Analyses of mammalian sera in enzyme-linked immunosorbent assays with different strains of Borrelia burgdorferi sensu lato.

    Science.gov (United States)

    Magnarelli, L A; Anderson, J F; Johnson, R C

    1995-04-01

    Blood samples were collected from cottontail rabbits (Sylvilagus floridanus), raccoons (Procyon lotor), white-footed mice (Peromyscus leucopus), and white-tailed deer (Odocoileus virginianus) between 1977 and 1991 in southern Connecticut and New York State (USA) and were tested for antibodies against eight strains of Borrelia burgdorferi sensu lato in enzyme-linked immunosorbent assays. Among these spirochetes were six strains of B. burgdorferi sensu stricto, one strain of B. garinii (=IP90) and a strain (IPF) in group VS461. Sera from each study group reacted positively to all strains having origins in North America and Eurasia. Assay sensitivities normally ranged between 85% and 100% for all study groups. The lowest sensitivity (66%) was noted when mouse sera were tested with B. garinii, an isolate from Ixodes persulcatus in the former Soviet Union. Differences in serum reactivity to various strains were noted for all study groups, but because of multiple shared antigens among the closely related spirochetes tested, the selection of a particular North American strain of B. burgdorferi sensu stricto did not appear to be a critical factor for optimal assay performance. Locally obtained strains of this bacterium are preferred as coating antigens for serologic testing because of their availability. PMID:8583632

  7. Establishment of a minor groove binder-probe based quantitative real time PCR to detect Borrelia burgdorferi sensu lato and differentiation of Borrelia spielmanii by ospA-specific conventional PCR

    Directory of Open Access Journals (Sweden)

    Strube Christina

    2010-08-01

    Full Text Available Abstract Background Borrelia burgdorferi sensu lato (sl, the causative agent of Lyme borreliosis, is transmitted by ticks of the genus Ixodes as vector. For identification of Borrelia infections in ticks a TaqMan™ minor groove binder (MGB probe-based quantitative real time PCR (qPCR was established targeting the 5S-23S intergenic spacer. Extension to a duplex qPCR included an Ixodes spp. positive control to verify successful DNA isolation. Besides qPCR, an ospA-specific conventional PCR for species-specific identification of B. spielmanii was established. Afterwards 1000 I. ricinus flagged in the city of Hanover, Germany, were investigated for B. burgdorferi sl infections followed by species identification. Furthermore, I. hexagonus ticks were investigated to proof applicability of the PCRs. Results Quantitative real time PCR (qPCR identifying B. burgdorferi sl in ticks was able to detect 1-10 copies per reaction. B. spielmanii ospA-specific conventional PCR was also highly specific and showed no cross reactions with the other tested Borrelia species. From 1000 hanoveranian ticks 24.3% were positive compared to only 7.4% positives by dark-field microscopy. Related to tick stage 1.7% larvae, 18.1% nymphs, and 34.6% adults were positive. The most frequent species was B. garinii, followed by B. afzelii, B. spielmanii, B. valaisiana and B. burgdorferi sensu stricto (ss. 70.6% of I. ricinus were mono-infected, whereas 28.0% and 1.4% were infected with two and three Borrelia species, respectively. From 232 I. hexagonus collected from hedgehogs in different sites of Germany, qPCR detected 5.7% to be infected with B. burgdorferi sl, which were identified as B. afzelii, B. garinii and B. spielmanii. Conclusions The evaluated qPCR to detect B. burgdorferi sl in Ixodes spp. is highly specific and sensitive. As a duplex qPCR including detection of Ixodes spp. DNA it is the first DNA based technique incorporating a control for successful DNA isolation from

  8. Whole-Genome Sequences of Borrelia bissettii, Borrelia valaisiana, and Borrelia spielmanii

    OpenAIRE

    Schutzer, Steven E.; Fraser-Liggett, Claire M.; Qiu, Wei-Gang; Kraiczy, Peter; Mongodin, Emmanuel F.; Dunn, John J.; Luft, Benjamin J.; Casjens, Sherwood R.

    2012-01-01

    It has been known for decades that human Lyme disease is caused by the three spirochete species Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii. Recently, Borrelia valaisiana, Borrelia spielmanii, and Borrelia bissettii have been associated with Lyme disease. We report the complete genome sequences of B. valaisiana VS116, B. spielmanii A14S, and B. bissettii DN127.

  9. Whole-Genome Sequences of Borrelia bissettii Borrelia valaisiana and Borrelia spielmanii

    Energy Technology Data Exchange (ETDEWEB)

    Schutzer S. E.; Dunn J.; Fraser-Liggett C. M.; Qiu W.-G.; Kraiczy P.; Mongodin E. F.; Luft B. J.; Casjens S. R.

    2012-01-01

    It has been known for decades that human Lyme disease is caused by the three spirochete species Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii. Recently, Borrelia valaisiana, Borrelia spielmanii, and Borrelia bissettii have been associated with Lyme disease. We report the complete genome sequences of B. valaisiana VS116, B. spielmanii A14S, and B. bissettii DN127.

  10. Probable late lyme disease: a variant manifestation of untreated Borrelia burgdorferi infection

    Directory of Open Access Journals (Sweden)

    Aucott John N

    2012-08-01

    Full Text Available Abstract Background Lyme disease, a bacterial infection with the tick-borne spirochete Borrelia burgdorferi, can cause early and late manifestations. The category of probable Lyme disease was recently added to the CDC surveillance case definition to describe patients with serologic evidence of exposure and physician-diagnosed disease in the absence of objective signs. We present a retrospective case series of 13 untreated patients with persistent symptoms of greater than 12 weeks duration who meet these criteria and suggest a label of ‘probable late Lyme disease’ for this presentation. Methods The sample for this analysis draws from a retrospective chart review of consecutive, adult patients presenting between August 2002 and August 2007 to the author (JA, an infectious disease specialist. Patients were included in the analysis if their current illness had lasted greater than or equal to 12 weeks duration at the time of evaluation. Results Probable late Lyme patients with positive IgG serology but no history of previous physician-documented Lyme disease or appropriate Lyme treatment were found to represent 6% of our heterogeneous sample presenting with ≥ 12 weeks of symptom duration. Patients experienced a range of symptoms including fatigue, widespread pain, and cognitive complaints. Approximately one-third of this subset reported a patient-observed rash at illness onset, with a similar proportion having been exposed to non-recommended antibiotics or glucocorticosteroid treatment for their initial disease. A clinically significant response to antibiotics treatment was noted in the majority of patients with probable late Lyme disease, although post-treatment symptom recurrence was common. Conclusions We suggest that patients with probable late Lyme disease share features with both confirmed late Lyme disease and post-treatment Lyme disease syndrome. Physicians should consider the recent inclusion of probable Lyme disease in the

  11. Borrelia burgdorferi CheD Promotes Various Functions in Chemotaxis and the Pathogenic Life Cycle of the Spirochete.

    Science.gov (United States)

    Moon, Ki Hwan; Hobbs, Gerry; Motaleb, M A

    2016-06-01

    Borrelia burgdorferi possesses a sophisticated chemotaxis signaling system; however, the roles of the majority of the chemotaxis proteins in the infectious life cycle have not yet been demonstrated. Specifically, the role of CheD during host colonization has not been demonstrated in any bacterium. Here, we systematically characterized the B. burgdorferi CheD homolog using genetics and biochemical and mouse-tick-mouse infection cycle studies. Bacillus subtilis CheD plays an important role in chemotaxis by deamidation of methyl-accepting chemotaxis protein receptors (MCPs) and by increasing the receptor kinase activity or enhancing CheC phosphatase activity, thereby regulating the levels of the CheY response regulator. Our biochemical analysis indicates that B. burgdorferi CheD significantly enhances CheX phosphatase activity by specifically interacting with the phosphatase. Moreover, CheD specifically binds two of the six MCPs, indicating that CheD may also modulate the receptor proteins. Although the motility of the cheD mutant cells was indistinguishable from that of the wild-type cells, the mutant did exhibit reduced chemotaxis. Importantly, the mutant showed significantly reduced infectivity in C3H/HeN mice via needle inoculation. Mouse-tick-mouse infection assays indicated that CheD is dispensable for acquisition or transmission of spirochetes; however, the viability of cheD mutants in ticks is marginally reduced compared to that of the wild-type or complemented cheD spirochetes. These data suggest that CheD plays an important role in the chemotaxis and pathogenesis of B. burgdorferi We propose potential connections between CheD, CheX, and MCPs and discuss how these interactions play critical roles during the infectious life cycle of the spirochete. PMID:27021244

  12. Lyme disease caused by Borrelia burgdorferi with two homeologous 16S rRNA genes: a case report.

    Science.gov (United States)

    Lee, Sin Hang

    2016-01-01

    Lyme disease (LD), the most common tick-borne disease in North America, is believed to be caused exclusively by Borrelia burgdorferi sensu stricto and is usually diagnosed by clinical evaluation and serologic assays. As reported previously in a peer-reviewed article, a 13-year-old boy living in the Northeast of the USA was initially diagnosed with LD based on evaluation of his clinical presentations and on serologic test results. The patient was treated with a course of oral doxycycline for 28 days, and the symptoms resolved. A year later, the boy developed a series of unusual symptoms and did not attend school for 1 year. A LD specialist reviewed the case and found the serologic test band patterns nondiagnostic of LD. The boy was admitted to a psychiatric hospital. After discharge from the psychiatric hospital, a polymerase chain reaction test performed in a winter month when the boy was 16 years old showed a low density of B. burgdorferi sensu lato in the blood of the patient, confirmed by partial 16S rRNA (ribosomal RNA) gene sequencing. Subsequent DNA sequencing analysis presented in this report demonstrated that the spirochete isolate was a novel strain of B. burgdorferi with two homeologous 16S rRNA genes, which has never been reported in the world literature. This case report shows that direct DNA sequencing is a valuable tool for reliable molecular diagnosis of Lyme and related borrelioses, as well as for studies of the diversity of the causative agents of LD because LD patients infected by a rare or novel borrelial variant may produce an antibody pattern that can be different from the pattern characteristic of an infection caused by a typical B. burgdorferi sensu stricto strain. PMID:27186082

  13. Insertion of fluorescent fatty acid probes into the outer membranes of the pathogenic spirochaetes Treponema pallidum and Borrelia burgdorferi.

    Science.gov (United States)

    Cox, D L; Radolf, J D

    2001-05-01

    The authors examined the ability of octadecanoyl (C(18)), hexadecanoyl (C(16)) and dodecanoyl (C(12)) fatty acid (FA) conjugates of 5-aminofluorescein (OAF, HAF and DAF, respectively) to insert into the outer membranes (OMs) of Treponema pallidum, Borrelia burgdorferi and Escherichia coli. Biophysical studies have demonstrated that these compounds stably insert into phospholipid bilayers with the acyl chain within the hydrophobic interior of the apical leaflet and the hydrophilic fluorescein moiety near the phospholipid head groups. Consistent with the known poor intrinsic permeability of the E. coli OM to hydrophobic compounds and surfactants, E. coli was not labelled with any of the FA probes. OAF inserted more readily into OMs of B. burgdorferi than into those of T. pallidum, although both organisms were completely labelled at concentrations at or below 2 microg ml(-1). Intact spirochaetes were labelled with OAF but not with antibodies against known periplasmic antigens, thereby confirming that the probe interacted exclusively with the spirochaetal OMs. Separate experiments in which organisms were cooled to 4 degrees C (i.e. below the OM phase-transition temperatures) indicated that labelling with OAF was due to insertion of the probe into the OMs. B. burgdorferi, but not T. pallidum, was labelled by relatively high concentrations of HAF and DAF. Taken as a whole, these findings support the prediction that the lack of lipopolysaccharide renders T. pallidum and B. burgdorferi OMs markedly more permeable to lipophilic compounds than their Gram-negative bacterial counterparts. The data also raise the intriguing possibility that these two pathogenic spirochaetes obtain long-chain FAs, nutrients they are unable to synthesize, by direct permeation of their OMs. PMID:11320119

  14. Lyme disease caused by Borrelia burgdorferi with two homeologous 16S rRNA genes: a case report

    Science.gov (United States)

    Lee, Sin Hang

    2016-01-01

    Lyme disease (LD), the most common tick-borne disease in North America, is believed to be caused exclusively by Borrelia burgdorferi sensu stricto and is usually diagnosed by clinical evaluation and serologic assays. As reported previously in a peer-reviewed article, a 13-year-old boy living in the Northeast of the USA was initially diagnosed with LD based on evaluation of his clinical presentations and on serologic test results. The patient was treated with a course of oral doxycycline for 28 days, and the symptoms resolved. A year later, the boy developed a series of unusual symptoms and did not attend school for 1 year. A LD specialist reviewed the case and found the serologic test band patterns nondiagnostic of LD. The boy was admitted to a psychiatric hospital. After discharge from the psychiatric hospital, a polymerase chain reaction test performed in a winter month when the boy was 16 years old showed a low density of B. burgdorferi sensu lato in the blood of the patient, confirmed by partial 16S rRNA (ribosomal RNA) gene sequencing. Subsequent DNA sequencing analysis presented in this report demonstrated that the spirochete isolate was a novel strain of B. burgdorferi with two homeologous 16S rRNA genes, which has never been reported in the world literature. This case report shows that direct DNA sequencing is a valuable tool for reliable molecular diagnosis of Lyme and related borrelioses, as well as for studies of the diversity of the causative agents of LD because LD patients infected by a rare or novel borrelial variant may produce an antibody pattern that can be different from the pattern characteristic of an infection caused by a typical B. burgdorferi sensu stricto strain. PMID:27186082

  15. 伯氏疏螺旋体膜蛋白BmpA研究进展%Progresses on Borrelia burgdorferi Membrance Protein A (BmpA)

    Institute of Scientific and Technical Information of China (English)

    宝福凯; 赖名耀; 张云波; 董坚; 赵桂萍; 陈明清; 柳爱华

    2012-01-01

    Lyme disease, a global health concern, is a zoonosis, which has been a serious threat to human. The spiroehete Borrelia burgdorferi that is transmitted by the bite of hard tick (Ixodidae) is the pathogen of Lyme disease. Borrelia burgdorferi contains many membrane proteins with immungenicity and pathogenicity. Recent researches show that BmpA is an dominant immune protein of Borrelia burgdorferi, a laminin-binding protein, and an arthritogennic factor. Research progresses of BmpA protein in biological function, Lyme arthritis pathogenesis and diagnosis of lyme disease are reviewed.%莱姆病是一种人兽共患病,已严重威胁人类健康,成全球公共卫生问题,引起全球关注.伯氏疏螺旋体是莱姆病病原体,通过蜱叮咬传播而引起莱姆病,其表面存在的膜蛋白具有免疫性和致病性.BmpA (Borreli burgdorferi membrance protein A)是伯氏疏螺旋体的主要抗原之一,为层粘连蛋白结合蛋白,是莱姆关节炎的重要致病因子,对蛋白功能、诊断应用和莱姆关节炎致病机理三方面的研究进展进行概述.

  16. Polymerase chain reaction in diagnosis of Borrelia burgdorferi infections and studies on taxonomic classification

    DEFF Research Database (Denmark)

    Lebech, Anne-Mette

    PCR assay for direct detection of B. burgdorferi DNA and to evaluate the diagnostic utility of PCR in clinical specimens from patients with Lyme borreliosis and (ii) to study the taxonomic classification of B. burgdorferi isolates and its implications for epidemiology and clinical presentation...... for PCR amplification and subsequent identification of B. burgdorferi specific sequences were established and used. For all assays the analytical sensitivity was a few genome copies using purified DNA as template. The efficacy of PCR was initially evaluated using tissue samples from experimentally...... infected gerbils in order to start with biological samples a priori known to contain B. burgdorferi. B. burgdorferi DNA was detectable in 88% of the specimens. Thus the diagnostic sensitivity of PCR was comparable to and even higher than in vitro culture. PCR was significantly more sensitive than a...

  17. Establishment of Multiple Locus Variable-number Tandem Repeat Analysis Assay for Genotyping of Borrelia burgdorferi sensu lato Detected in China

    Institute of Scientific and Technical Information of China (English)

    ZHOU Xin; HOU Xue Xia; GENG Zhen; ZHAO Rui; WAN Kang Lin; HAO Qin

    2014-01-01

    Objective Human Lyme Borreliosis (LB), which is caused by Borrelia burgdorferi sensu lato (B. burgdorferi), has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multiple locus variable-number tandem repeat (VNTR) analysis (MLVA) assay for the genotyping of Borrelia burgdorferi strains detected in China. Methods B. garinii PBi complete 904.246 kb chromosome and two plasmids (cp26 and lp54) were screened by using Tandem Repeats Finder program for getting potential VNTR loci, the potential VNTR loci were analyzed and identified with PCR and the VNTR loci data were analyzed and MLVA clustering tree were constrcted by using the categorical coefficient and the unweighted pair-group method with arithmetic means (UPGMA). Results We identified 5 new VNTR loci through analyzing 47 potential VNTR loci. We used the MLVA protocol to analyse 101 B. burgdorferi strains detected in China and finally identified 51 unique genotypes in 4 major clusters including B. burgdorferi sensu stricto (B.b.s.s), B. garinii, B. afzelii, and B. valaisiana, consistent with the current MLSA phylogeny studies. The allele numbers of VNTR-1, VNTR-2, VNTR-3, VNTR-4, and VNTR-5 were 7, 3, 9, 7, and 6. The Hunter-Gaston index (HGI) of five VNTR loci were 0.79, 0.22, 0.77, 0.71, and 0.67, respectively. The combined HGI of five VNTR loci was 0.96. Clustering of the strains of Xinjiang, Inner Mongolia and Heilongjiang was confirmed, and this situation was consistent with the close geographical distribution of those provinces. Conclusion The MLVA protocol esytablished in this study is easy and can show strains’ phylogenetic relationships to distinguish the strains of Borrelia species. It is useful for further phylogenetic and epidemiological analyses of Borrelia strains.

  18. High Prevalence of Granulocytic Ehrlichiae and Borrelia burgdorferi Sensu Lato in Ixodes ricinus Ticks from Bulgaria

    OpenAIRE

    Christova, Iva; Schouls, Leo; van de Pol, Ingrid; Park, Jinho; Panayotov, Stefan; Lefterova, Viktoria; Kantardjiev, Todor; Dumler, J. Stephen

    2001-01-01

    Bulgarian Ixodes ricinus ticks were examined for Ehrlichia and Borrelia coinfection: 34 and 32% of adult ticks and at least 2 and 10% of nymphs were positive for these infections, respectively. Coinfections and dual or triple Borrelia infections were frequent, although Ehrlichia phagocytophila heterogeneity was minimal. Multiple tick-borne bacteria coexist in I. ricinus ticks in southeastern Europe.

  19. Response to Esteve-Gassent et al.: flaB sequences obtained from Texas PCR products are identical to the positive control strain Borrelia burgdorferi B31.

    Science.gov (United States)

    Norris, Steven J; Barbour, Alan G; Fish, Durland; Diuk-Wasser, Maria A

    2015-01-01

    Feria-Arroyo et al. had reported previously that, based on PCR analysis, 45% of Ixodes scapularis ticks collected in Texas and Mexico were infected with the Lyme disease spirochete Borrelia burgdorferi (Parasit. Vectors 2014, 7:199). However, our analyses of their initial data (Parasit. Vectors 2014, 7:467) and a recent response by Esteve-Gassent et al. (Parasit. Vectors 2015, 8:129) provide evidence that the positive PCR results obtained from both ribosomal RNA intergenic sequences and the flagellin gene flaB are highly likely due to contamination by the B. burgdorferi B31 positive control strain. PMID:26050617

  20. Minimal role of eastern fence lizards in Borrelia burgdorferi transmission in central New Jersey oak/pine woodlands

    Science.gov (United States)

    Rulison, Eric L.; Kerr, Kaetlyn T; Dyer, Megan C; Han, Seungeun; Burke, Russell L.; Tsao, Jean I.; Ginsberg, Howard S.

    2014-01-01

    The Eastern fence lizard, Sceloporus undulatus, is widely distributed in eastern and central North America, ranging through areas with high levels of Lyme disease, as well as areas where Lyme disease is rare or absent. We studied the potential role of S. undulatus in transmission dynamics of Lyme spirochetes by sampling ticks from a variety of natural hosts at field sites in central New Jersey, and by testing the reservoir competence of S. undulatus for Borrelia burgdorferi in the laboratory. The infestation rate of ticks on fence lizards was extremely low (proportion infested = 0.087, n = 23) compared to that on white footed mice and other small mammals (proportion infested = 0.53, n = 140). Of 159 nymphs that had fed as larvae on lizards that had previously been exposed to infected nymphs, none was infected with B. burgdorferi, compared with 79.9% of 209 nymphs that had fed as larvae on infected control mice. Simulations suggest that changes in the numbers of fence lizards in a natural habitat would have little effect on the infection rate of nymphal ticks with Lyme spirochetes. We conclude that in central New Jersey S. undulatus plays a minimal role in the enzootic transmission cycle of Lyme spirochetes.

  1. Treponema pallidum Lipoprotein TP0435 Expressed in Borrelia burgdorferi Produces Multiple Surface/Periplasmic Isoforms and mediates Adherence.

    Science.gov (United States)

    Chan, Kamfai; Nasereddin, Thayer; Alter, Laura; Centurion-Lara, Arturo; Giacani, Lorenzo; Parveen, Nikhat

    2016-01-01

    The ability of Treponema pallidum, the syphilis spirochete to colonize various tissues requires the presence of surface-exposed adhesins that have been difficult to identify due to the inability to culture and genetically manipulate T. pallidum. Using a Borrelia burgdorferi-based heterologous system and gain-in-function approach, we show for the first time that a highly immunogenic lipoprotein TP0435 can be differentially processed into multiple isoforms with one variant stochastically displayed on the spirochete surface. TP0435 was previously believed to be exclusively located in T. pallidum periplasm. Furthermore, non-adherent B. burgdorferi strain expressing TP0435 acquires the ability to bind to a variety of host cells including placental cells and exhibits slow opsonophagocytosis in vitro similar to poor ex vivo phagocytosis of T. pallidum by host macrophages reported previously. This phenomenon of production of both surface and periplasmic immunogenic lipoprotein isoforms has possible implications in immune evasion of the obligate pathogen T. pallidum during infection. PMID:27161310

  2. Analysis of an ordered, comprehensive STM mutant library in infectious Borrelia burgdorferi: insights into the genes required for mouse infectivity.

    Directory of Open Access Journals (Sweden)

    Tao Lin

    Full Text Available The identification of genes important in the pathogenesis of Lyme disease Borrelia has been hampered by exceedingly low transformation rates in low-passage, infectious organisms. Using the infectious, moderately transformable B. burgdorferi derivative 5A18NP1 and signature-tagged versions of the Himar1 transposon vector pGKT, we have constructed a defined transposon library for the efficient genome-wide investigation of genes required for wild-type pathogenesis, in vitro growth, physiology, morphology, and plasmid replication. To facilitate analysis, the insertion sites of 4,479 transposon mutants were determined by sequencing. The transposon insertions were widely distributed across the entire B. burgdorferi genome, with an average of 2.68 unique insertion sites per kb DNA. The 10 linear plasmids and 9 circular plasmids had insertions in 33 to 100 percent of their predicted genes. In contrast, only 35% of genes in the 910 kb linear chromosome had incapacitating insertions; therefore, the remaining 601 chromosomal genes may represent essential gene candidates. In initial signature-tagged mutagenesis (STM analyses, 434 mutants were examined at multiple tissue sites for infectivity in mice using a semi-quantitative, Luminex-based DNA detection method. Examples of genes found to be important in mouse infectivity included those involved in motility, chemotaxis, the phosphoenolpyruvate phosphotransferase system, and other transporters, as well as putative plasmid maintenance genes. Availability of this ordered STM library and a high-throughput screening method is expected to lead to efficient assessment of the roles of B. burgdorferi genes in the infectious cycle and pathogenesis of Lyme disease.

  3. Development and validation of a FACS-based lipoprotein localization screen in the Lyme disease spirochete Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Slusser Joyce G

    2010-11-01

    Full Text Available Abstract Background In our previous studies on lipoprotein secretion in the Lyme disease spirochete Borrelia burgdorferi, we used monomeric red fluorescent protein 1 (mRFP1 fused to specifically mutated outer surface protein A (OspA N-terminal lipopeptides to gather first insights into lipoprotein sorting determinants. OspA:mRFP1 fusions could be detected by epifluorescence microscopy both in the periplasm and on the bacterial surface. To build on these findings and to complement the prior targeted mutagenesis approach, we set out to develop a screen to probe a random mutagenesis expression library for mutants expressing differentially localized lipoproteins. Results A Glu-Asp codon pair in the inner membrane-localized OspA20:mRFP1 fusion was chosen for mutagenesis since the two negative charges were previously shown to define the phenotype. A library of random mutants in the two codons was generated and expressed in B. burgdorferi. In situ surface proteolysis combined with fluorescence activated cell sorting (FACS was then used to screen for viable spirochetes expressing alternative subsurface OspA:mRFP1 fusions. Analysis of 93 clones randomly picked from a sorted cell population identified a total of 43 distinct mutants. Protein localization assays indicated a significant enrichment in the selected subsurface phenotype. Interestingly, a majority of the subsurface mutant proteins localized to the outer membrane, indicating their impairment in "flipping" through the outer membrane to the spirochetal surface. OspA20:mRFP1 remained the protein most restricted to the inner membrane. Conclusions Together, these results validate this FACS-based screen for lipoprotein localization and suggest a rather specific inner membrane retention mechanism involving membrane anchor-proximal negative charge patches in this model B. burgdorferi lipoprotein system.

  4. An RND-type efflux system in Borrelia burgdorferi is involved in virulence and resistance to antimicrobial compounds.

    Directory of Open Access Journals (Sweden)

    Ignas Bunikis

    2008-02-01

    Full Text Available Borrelia burgdorferi is remarkable for its ability to thrive in widely different environments due to its ability to infect various organisms. In comparison to enteric Gram-negative bacteria, these spirochetes have only a few transmembrane proteins some of which are thought to play a role in solute and nutrient uptake and excretion of toxic substances. Here, we have identified an outer membrane protein, BesC, which is part of a putative export system comprising the components BesA, BesB and BesC. We show that BesC, a TolC homolog, forms channels in planar lipid bilayers and is involved in antibiotic resistance. A besC knockout was unable to establish infection in mice, signifying the importance of this outer membrane channel in the mammalian host. The biophysical properties of BesC could be explained by a model based on the channel-tunnel structure. We have also generated a structural model of the efflux apparatus showing the putative spatial orientation of BesC with respect to the AcrAB homologs BesAB. We believe that our findings will be helpful in unraveling the pathogenic mechanisms of borreliae as well as in developing novel therapeutic agents aiming to block the function of this secretion apparatus.

  5. Isolation of DNA after Extraction of RNA To Detect the Presence of Borrelia burgdorferi and Expression of Host Cellular Genes from the Same Tissue Sample

    OpenAIRE

    Amemiya, Kei; Schaefer, Henry; Pachner, Andrew R.

    1999-01-01

    We are investigating the neuropathogenesis of Lyme disease caused by Borrelia burgdorferi in a nonhuman primate model. In the past, two separate pieces of tissue had to be used when both analyzing for the presence of the spirochete and examining the host response to infection. We have modified a procedure to purify DNA from the same sample after the extraction of RNA. The remaining material containing the DNA was precipitated, and residual organic reagent was removed prior to deproteinization...

  6. Infectivity of the Highly Transformable BBE02− lp56− Mutant of Borrelia burgdorferi, the Lyme Disease Spirochete, via Ticks

    OpenAIRE

    Jacobs, Mary B.; Norris, Steven J; Kathrine M Phillippi-Falkenstein; Philipp, Mario T.

    2006-01-01

    Infectious Borrelia burgdorferi strains that have increased transformability with the shuttle vector pBSV2 were recently constructed by inactivating the gene encoding BBE02, a putative restriction-modification gene product expressed by the linear plasmid lp25 (Kawabata et al., Infect. Immun. 72:7147-7154, 2004). The absence of the linear plasmid lp56, which carries another putative restriction-modification gene, further enhanced transformation rates. The infectivity of these mutants was asses...

  7. Prävalenz von hämotrophen Mycoplasma spp., Bartonella spp., Anaplasma phagocytophilum und Borrelia burgdorferi bei Katzen im Raum Berlin/Brandenburg

    OpenAIRE

    Morgenthal, Dinah

    2014-01-01

    Arthropod-borne infectious agents in the cat include Bartonella (B.) henselae, Bartonella (B.) clarridgeiae, Bartonella (B.) quintana, Anaplasma (A.) phagocytophilum and Borrelia (B.) burgdorferi. Moreover infections with Mycoplasma (M.) haemofelis, Candidatus Mycoplasma (C. M.) turicensis and Candidatus Mycoplasma (C. M.) haemominutum occur in the cat. The cat flea (Ctenocephalides felis) is supposed to be the main vector for Mycoplasma spp.; moreover, transmission via blood transfusion and ...

  8. Subjective health complaints are not associated with tick bites or antibodies to Borrelia burgdorferi sensu lato in blood donors in western Norway: a cross-sectional study

    OpenAIRE

    2015-01-01

    Background There is controversy about chronic health consequences of tick-borne infections, especially Lyme borreliosis. This study aims to assess whether general function, physical fitness and subjective health complaints are associated with tick bites or antibodies to Borrelia burgdorferi sensu lato in blood donors. Methods Sera from 1,213 blood donors at four different blood banks in Sogn and Fjordane county in western Norway were obtained during January to June 2010, and analysed for s...

  9. Subjective health complaints are not associated with tick bites or antibodies to Borrelia burgdorferi sensu lato in blood donors in western Norway: a cross-sectional study

    OpenAIRE

    Hjetland, Reidar; Reiso, Harald; Ihlebæk, Camilla; Nilsen, Roy M; Grude, Nils; Ulvestad, Elling

    2015-01-01

    Background There is controversy about chronic health consequences of tick-borne infections, especially Lyme borreliosis. This study aims to assess whether general function, physical fitness and subjective health complaints are associated with tick bites or antibodies to Borrelia burgdorferi sensu lato in blood donors. Methods Sera from 1,213 blood donors at four different blood banks in Sogn and Fjordane county in western Norway were obtained during January to June 2010, and analysed for spec...

  10. Subjective health complaints are not associated with tick bites or antibodies to Borrelia burgdorferi sensu lato in blood donors in western Norway: a cross-sectional study

    OpenAIRE

    Hjetland, Reidar; Reiso, Harald; Ihlebæk, Camilla; Nilsen, Roy Miodini; Grude, Nils; Ulvestad, Elling

    2015-01-01

    Background: There is controversy about chronic health consequences of tick-borne infections, especially Lyme borreliosis. This study aims to assess whether general function, physical fitness and subjective health complaints are associated with tick bites or antibodies to Borrelia burgdorferi sensu lato in blood donors. Methods: Sera from 1,213 blood donors at four different blood banks in Sogn and Fjordane county in western Norway were obtained during January to June 2010, and analysed for...

  11. Tick saliva affects both proliferation and distribution of Borrelia burgdorferi spirochetes in mouse organs and increases transmission of spirochetes to ticks

    Czech Academy of Sciences Publication Activity Database

    Horká, Helena; Černá-Kýčková, Kateřina; Skallová, Anna; Kopecký, Jan

    2009-01-01

    Roč. 299, č. 5 (2009), s. 373-380. ISSN 1438-4221 R&D Projects: GA AV ČR IAA600960811; GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia burgdorferi * Ixodes ricinus * saliva-activated transmission * real -time PCR Subject RIV: EC - Immunology Impact factor: 2.802, year: 2009

  12. The Nucleotide Excision Repair System of Borrelia burgdorferi Is the Sole Pathway Involved in Repair of DNA Damage by UV Light

    OpenAIRE

    Hardy, Pierre-Olivier; Chaconas, George

    2013-01-01

    To survive and avoid accumulation of mutations caused by DNA damage, the genomes of prokaryotes encode a variety of DNA repair pathways most well characterized in Escherichia coli. Some of these are required for the infectivity of various pathogens. In this study, the importance of 25 DNA repair/recombination genes for Borrelia burgdorferi survival to UV-induced DNA damage was assessed. In contrast to E. coli, where 15 of these genes have an effect on survival of UV irradiation, disruption of...

  13. Production of reactive oxygen species and expression of inducible nitric oxide synthase in rat isolated Kupffer cells stimulated by Leptospira interrogans and Borrelia burgdorferi

    Institute of Scientific and Technical Information of China (English)

    Antonella Marangoni; Silvia Accardo; Rita Aldini; Massimo Guardigli; Francesca Cavrini; Vittorio Sambri; Marco Montagnani; Aldo Roda; Roberto Cevenini

    2006-01-01

    AIM: To evaluate the production of reactive oxygen species (ROS) and the expression of indudble nitric oxide synthase (iNOS) in rat isolated Kupffer cells (KCs) stimulated by Leptospira interrogans and Borrelia burgdorferi.METHODS: Rat Kupffer cells were separated by perfusion of the liver with 0.05% collagenase, and purified by Percoll gradients. Purified Kupffer cells were tested in vitro with alive L.interogans and B. burgdorferi preparations. The production of ROS was determined by chemiluminescence, whereas iNOS protein expression was evaluated by Western blot assay using anti-iNOS antibodies.RESULTS: B. burgdorferi and to a less extent L. interrogans induced ROS production with a peak 35 min after infection. The chemiluminescence signal progressively diminished and was undetectable by 180 min of incubation. Leptospirae and borreliae induced an increased iNOS expression in Kupffer cells that peaked at 6 hours and was still evident 22 h after infection.CONCLUSION: Both genera of spirochetes induced ROS and iNOS production in rat Kupffer cells. Since the cause of liver damage both in leptospiral as well as in borrelial infections are still unknown, we suggest that leptospira and borrelia damage of the liver can be initially mediated by oxygen radicals, and is then maintained at least in part by nitric oxide.

  14. Efficacy of sarolaner in the prevention of Borrelia burgdorferi and Anaplasma phagocytophilum transmission from infected Ixodes scapularis to dogs.

    Science.gov (United States)

    Honsberger, Nicole A; Six, Robert H; Heinz, Thomas J; Weber, Angela; Mahabir, Sean P; Berg, Thomas C

    2016-05-30

    The efficacy of sarolaner (Simparica™, Zoetis) to prevent transmission primarily of Borrelia burgdorferi and secondarily of Anaplasma phagocytophilum from infected wild-caught Ixodes scapularis to dogs was evaluated in a placebo-controlled laboratory study. Twenty-four purpose-bred laboratory Beagles seronegative for B. burgdorferi and A. phagocytophilum antibodies were allocated randomly to one of three treatment groups: placebo administered orally on Days 0 and 7, or sarolaner at 2mg/kg administered orally on Day 0 (28 days prior to tick infestation) or on Day 7 (21 days prior to tick infestation). On Day 28, each dog was infested with approximately 25 female and 25 male wild caught adult I. scapularis that were determined to have prevalence of 57% for B. burgdorferi and 6.7% for A. phagocytophilum by PCR. In situ tick counts were conducted on Days 29 and 30. On Day 33, all ticks were counted and removed. Acaricidal efficacy was calculated based on the reduction of geometric mean live tick counts in the sarolaner-treated groups compared to the placebo-treated group for each tick count. Blood samples collected from each dog on Days 27, 49, 63, 77, 91 and 104 were tested for the presence of B. burgdorferi and A. phagocytophilum antibodies using the SNAP(®) 4Dx(®) Plus Test, and quantitatively assayed for B. burgdorferi antibodies using an ELISA test. Skin biopsies collected on Day 104 were tested for the presence of B. burgdorferi by bacterial culture and PCR. Geometric mean live tick counts for placebo-treated dogs were 14.8, 12.8, and 19.1 on Days 29, 30, and 33, respectively. The percent reductions in mean live tick counts at 1, 2, and 5 days after infestation were 86.3%, 100%, and 100% for the group treated with sarolaner 21 days prior to infestation, and 90.9%, 97.1%, and 100% for the group treated with sarolaner 28 days prior to infestation. Geometric mean live tick counts for both sarolaner-treated groups were significantly lower than those for the

  15. Detection and identification of Anaplasma phagocytophilum, Borrelia burgdorferi, and Rickettsia helvetica in Danish Ixodes ricinus ticks

    DEFF Research Database (Denmark)

    Skarphédinsson, Sigurdur; Lyholm, Birgitte Fjendbo; Ljungberg, Marianne;

    2007-01-01

    % of adult ticks. The difference in prevalence between Anaplasma and Borrelia in adult ticks supports the idea that their maintenance cycles in nature may be different. Ticks were also infected with Rickettsia helvetica. Our study indicates that A. phagocytophilum prevalence in ticks in Denmark is as...

  16. Blood feeding on large grazers affects the transmission of Borrelia burgdorferi sensu lato by Ixodes ricinus

    NARCIS (Netherlands)

    Pacilly, F.C.A.; Benning, M.E.; Jacobs, F.; Leidekker, J.; Sprong, H.; Wieren, van S.E.; Takken, W.

    2014-01-01

    The presence of Ixodes ricinus and their associated Borrelia infections on large grazers was investigated. Carcases of freshly shot red deer, mouflon and wild boar were examined for the presence of any stage of I. ricinus. Questing ticks were collected from locations where red deer and wild boar are

  17. Lipoproteins of Borrelia burgdorferi and Treponema pallidum activate cachectin/tumor necrosis factor synthesis. Analysis using a CAT reporter construct.

    Science.gov (United States)

    Radolf, J D; Norgard, M V; Brandt, M E; Isaacs, R D; Thompson, P A; Beutler, B

    1991-09-15

    Lipoproteins from two pathogenic spirochetes (Borrelia burgdorferi and Treponema pallidum) induced the biosynthesis of TNF in murine macrophages and in permanently transformed macrophages of the cell line RAW 264.7. Induction was studied by measuring the secretion of biologically active TNF and by measuring the activity of the reporter enzyme chloramphenicol acetyltransferase (CAT) produced within macrophages transfected with an endotoxin-responsive CAT construct. Several lines of evidence indicated that the induction of TNF and CAT was attributable to the spirochete lipoproteins rather than to contaminating or endogenous LPS: 1) the dose response curves observed for the lipoproteins were markedly different from those obtained with LPS; 2) lipoprotein-mediated activation was unaffected by amounts of polymyxin B that completely neutralized the induction of TNF and CAT by LPS, 3) low concentrations of the lipoproteins induced TNF in macrophages from endotoxin-unresponsive C3H/HeJ mice as effectively as in macrophages from normal C3H/HeN mice, and 4) isolated spirochete lipoproteins, but not a non-lipoprotein immunogen, were potent inducers of CAT in the transformed macrophages. Moreover, LPS was not detected in the B. burgdorferi lipoprotein mixtures by Limulus amebocyte lysate assay. Proteolytic digestion of the intact bacterial protein preparations only modestly diminished their ability to activate the cells, suggesting that small lipopeptides comprise the biologically active portions of the molecules, as is the case with the murein lipoprotein of Escherichia coli. Through their ability to induce TNF production by macrophages, spirochete lipoproteins may play important roles in the development of the local inflammatory changes and the systemic manifestations that characterize syphilis and Lyme disease. PMID:1890308

  18. BB0844, an RpoS-Regulated Protein, Is Dispensable for Borrelia burgdorferi Infectivity and Maintenance in the Mouse-Tick Infectious Cycle▿

    Science.gov (United States)

    Banik, Sukalyani; Terekhova, Darya; Iyer, Radha; Pappas, Christopher J.; Caimano, Melissa J.; Radolf, Justin D.; Schwartz, Ira

    2011-01-01

    The genome of Borrelia burgdorferi, the causative agent of Lyme disease, is comprised of a large linear chromosome and numerous smaller linear and circular plasmids. B. burgdorferi exhibits substantial genomic variation, and previous studies revealed genotype-specific variation at the right chromosomal telomere. A correlation has also been established between genotype and invasiveness. The correlation between chromosome length and genotype and between genotype and invasiveness suggested that a gene(s) at the right chromosome telomere may be required for virulence. Of particular interest was bb0844, an RpoS-regulated gene at the right telomere, the expression of which is induced when the spirochete undergoes adaptation to the mammalian host. The structure of the right chromosomal telomere was examined in 53 B. burgdorferi clinical isolates of various genotypes. Four distinct patterns were observed for bb0844: (i) chromosomal localization, (ii) plasmid localization, (iii) presence on both chromosome and plasmid, and (iv) complete absence. These patterns correlated with the B. burgdorferi genotype. On the basis of available sequence data, we propose a mechanism for the genomic rearrangements that accounts for the variability in bb0844 genomic localization. To further explore the role of BB0844 in the spirochete life cycle, a bb0844 deletion mutant was constructed by allelic exchange, and the viability of wild-type and bb0844 deletion mutants was examined in an experimental mouse-tick infection model. The bb0844 mutant was fully infectious in C3H/HeJ mice by either needle inoculation or tick transmission with B. burgdorferi-infected Ixodes scapularis larvae. Naïve larval ticks acquired both wild-type and mutant spirochetes with equal efficiency from B. burgdorferi-infected mice. The results demonstrate that BB0844 is not required for spirochete viability, pathogenicity, or maintenance in the tick vector or the mammalian host. At present, a defined role for BB0844 in B

  19. Central role of the Holliday junction helicase RuvAB in vlsE recombination and infectivity of Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Tao Lin

    2009-12-01

    Full Text Available Antigenic variation plays a vital role in the pathogenesis of many infectious bacteria and protozoa including Borrelia burgdorferi, the causative agent of Lyme disease. VlsE, a 35 kDa surface-exposed lipoprotein, undergoes antigenic variation during B. burgdorferi infection of mammalian hosts, and is believed to be a critical mechanism by which the spirochetes evade immune clearance. Random, segmental recombination between the expressed vlsE gene and adjacent vls silent cassettes generates a large number of different VlsE variants within the infected host. Although the occurrence and importance of vlsE sequence variation is well established, little is known about the biological mechanism of vlsE recombination. To identify factors important in antigenic variation and vlsE recombination, we screened transposon mutants of genes known to be involved in DNA recombination and repair for their effects on infectivity and vlsE recombination. Several mutants, including those in BB0023 (ruvA, BB0022 (ruvB, BB0797 (mutS, and BB0098 (mutS-II, showed reduced infectivity in immunocompetent C3H/HeN mice. Mutants in ruvA and ruvB exhibited greatly reduced rates of vlsE recombination in C3H/HeN mice, as determined by restriction fragment polymorphism (RFLP screening and DNA sequence analysis. In severe combined immunodeficiency (C3H/scid mice, the ruvA mutant retained full infectivity; however, all recovered clones retained the 'parental' vlsE sequence, consistent with low rates of vlsE recombination. These results suggest that the reduced infectivity of ruvA and ruvB mutants is the result of ineffective vlsE recombination and underscores the important role that vlsE recombination plays in immune evasion. Based on functional studies in other organisms, the RuvAB complex of B. burgdorferi may promote branch migration of Holliday junctions during vlsE recombination. Our findings are consistent with those in the accompanying article by Dresser et al., and together

  20. ER-Coordinated Activities of Rab22a and Rab5a Drive Phagosomal Compaction and Intracellular Processing of Borrelia burgdorferi by Macrophages

    Directory of Open Access Journals (Sweden)

    Xenia Naj

    2015-09-01

    Full Text Available Borrelia burgdorferi is the causative agent of Lyme disease, a multisystemic disorder affecting the skin, joints, and nervous system. Macrophages and dendritic cells counteract Borrelia dissemination through internalization and degradation of spirochetes. We now show that Borrelia internalization by primary human macrophages involves uptake and compaction into Rab22a-positive phagosomes that are in close contact with Rab5a-positive vesicles. Compaction of borreliae involves membrane extrusion from phagosomes, is driven by Rab22a and Rab5a activity, and is coordinated by ER tubules forming contact sites of Rab22a phagosomes with Rab5a vesicles. Importantly, Rab22a and Rab5a depletion leads to reduced localization to lysosomes and to increased intracellular survival of spirochetes. These data show that Rab22a- and Rab5a-driven phagosomal uptake is a crucial step in the vesicular cascade that leads to elimination of spirochetes by macrophages. Rab22a and Rab5a thus present as potential molecular targets for the modulation of intracellular processing of borreliae in human immune cells.

  1. Updates on Borrelia burgdorferi sensu lato complex with respect to public health

    Czech Academy of Sciences Publication Activity Database

    Rudenko, Natalia; Golovchenko, Maryna; Grubhoffer, Libor; Oliver, J. H., Jr.

    2011-01-01

    Roč. 2, č. 3 (2011), s. 123-128. ISSN 1877-959X R&D Projects: GA MŠk(CZ) LC06009; GA ČR GA206/09/1782 Institutional research plan: CEZ:AV0Z60220518 Keywords : Spirochetes * Borrelia * Genetic diversity * Lyme borreliosis * Distribution * New species Subject RIV: EE - Microbiology, Virology Impact factor: 2.370, year: 2011

  2. Growth inhibiting activity of volatile oil from Cistus creticus L. against Borrelia burgdorferi s.s. in vitro.

    Science.gov (United States)

    Hutschenreuther, A; Birkemeyer, C; Grötzinger, K; Straubinger, R K; Rauwald, H W

    2010-04-01

    Borreliosis patients from self-help groups reported considerable pain relief after ingestion of Cistus creticus leaf preparations. C. creticus leaf extracts of different polarities such as aqueous, ethyl acetate, hexane extracts as well as the volatile oil fraction obtained by steam distillation were tested for their antibacterial activity against Borrelia burgdorferi sensu stricto (Bbss) in vitro using the antibiotic amoxicilline as standard and polysorbate 80 as solubilizer for lipophilic extracts. Comparison of the four plant preparations shows that the volatile oil exerts the strongest growth inhibitory effect. Even concentrations of 0.02% (w/v) volatile oil in cultivation media reduced the total number of bacteria to 2% in comparison to a growth control after an eight-day cultivation period. While the aqueous extract did not reduce bacterial growth, incubation with hexane and ethyl acetate extracts clearly inhibited microbial growth. The main volatile components of the three active extracts tested were analyzed by GC-MS. The number of different labdane-type diterpenes as well as the total relative amount of diterpenes in the samples tested was highest in the essential oil of C. creticus. Identification of ten different volatile labdane-type diterpenes was assigned to the essential oil of C. creticus. Among these, manoyl oxide, 13-epi-manoyl oxide, 3-acetoxy-manoyl oxide and the monoterpene carvacrol were determined to be major constituents, accompanied by minor amounts of 3-hydroxy-manoyl oxide, all of which are known to exert antimicrobial activity. PMID:20432627

  3. Detección de anticuerpos contra Borrelia burgdorferi e identificación de garrapatas ixodidas en Piura Y Amazonas, Perú

    Directory of Open Access Journals (Sweden)

    Martha Glenny A

    2004-01-01

    Full Text Available Objetivos: Detectar anticuerpos IgG/IgM contra Borrelia burgdorferi en población general, procedentes de los departamentos de Piura y Amazonas e identificar especies de garrapatas probablemente incriminadas en la transmisión de la enfermedad de Lyme. Material y Métodos: Entre agosto del año 2001 y junio de 2002, se colectaron muestras de sangre de 232 pobladores procedentes de ocho localidades del Departamento de Piura y 12 del Departamento de Amazonas, para evaluar mediante ELISA Captia™ Lyme IgG/IgM (Trinity biotech la presencia de anticuerpos contra Borrelia burgdorferi. Además, se colectaron garrapatas en animales domésticos por búsqueda directa. Resultados: Se detectó seropositividad en 9,9 % de los sueros evaluados. Asimismo, de 433 garrapatas colectadas se identificaron los géneros: Ixodes (5,5%, Amblyomma (18,0%, Rhipicephalus (23,5%, Anocentor (31,1% y Boophilus (21,7%. Conclusiones: Existen personas seropositivas por Borrelia en Piura y Amazonas, coincidiendo con los hallazgos realizados en Sapillica en el año 1992, además se detectó la presencia de garrapatas del género Ixodes en Piura.

  4. A Drug Combination Screen Identifies Drugs Active against Amoxicillin-induced Round Bodies of Borrelia burgdorferi Persisters from an FDA Drug Library

    Directory of Open Access Journals (Sweden)

    Jie eFeng

    2016-05-01

    Full Text Available Although currently recommended antibiotics for Lyme disease such as doxycycline or amoxicillin cure the majority of the patients, about 10-20% of patients treated for Lyme disease may experience lingering symptoms including fatigue, pain, or joint and muscle aches. Under stress conditions such as starvation or antibiotic exposure, Borrelia burgdorferi can develop round body forms, which are a type of persister bacteria that are not killed by current Lyme antibiotics. To identify more effective drugs that are active against the round bodies of B. burgdorferi, we established a round body persister model induced by amoxicillin and screened the Food and Drug Administration (FDA drug library consisting of 1581 drug compounds and also 22 drug combinations using the SYBR Green I/propidium iodide (PI viability assay. We identified 23 drug candidates that have higher activity against the round bodies of B. burgdorferi than either amoxicillin or doxycycline. Eleven of these scored better than metronidazole and tinidazole which have been previously described to be active against round bodies. While some drug candidates such as daptomycin and clofazimine overlapped with a previous screen against stationary phase B. burgdorferi persisters, additional drug candidates active against round bodies we identified include artemisinin, ciprofloxacin, nifuroxime, fosfomycin, chlortetracycline, sulfacetamide, sulfamethoxypyridazine and sulfathiozole. Two triple drug combinations had the highest activity against round bodies and stationary phase B. burgdorferi persisters: artemisinin/cefoperazone/doxycycline and sulfachlorpyridazine/daptomycin/doxycycline. These findings confirm and extend previous findings that certain drug combinations have superior activity against B. burgdorferi persisters in vitro, even if pre-treated with amoxicillin. These findings may have implications for improved treatment of Lyme disease.

  5. A Drug Combination Screen Identifies Drugs Active against Amoxicillin-Induced Round Bodies of In Vitro Borrelia burgdorferi Persisters from an FDA Drug Library

    Science.gov (United States)

    Feng, Jie; Shi, Wanliang; Zhang, Shuo; Sullivan, David; Auwaerter, Paul G.; Zhang, Ying

    2016-01-01

    Although currently recommended antibiotics for Lyme disease such as doxycycline or amoxicillin cure the majority of the patients, about 10–20% of patients treated for Lyme disease may experience lingering symptoms including fatigue, pain, or joint and muscle aches. Under experimental stress conditions such as starvation or antibiotic exposure, Borrelia burgdorferi can develop round body forms, which are a type of persister bacteria that appear resistant in vitro to customary first-line antibiotics for Lyme disease. To identify more effective drugs with activity against the round body form of B. burgdorferi, we established a round body persister model induced by exposure to amoxicillin (50 μg/ml) and then screened the Food and Drug Administration drug library consisting of 1581 drug compounds and also 22 drug combinations using the SYBR Green I/propidium iodide viability assay. We identified 23 drug candidates that have higher activity against the round bodies of B. burgdorferi than either amoxicillin or doxycycline. Eleven individual drugs scored better than metronidazole and tinidazole which have been previously described to be active against round bodies. In this amoxicillin-induced round body model, some drug candidates such as daptomycin and clofazimine also displayed enhanced activity which was similar to a previous screen against stationary phase B. burgdorferi persisters not exposure to amoxicillin. Additional candidate drugs active against round bodies identified include artemisinin, ciprofloxacin, nifuroxime, fosfomycin, chlortetracycline, sulfacetamide, sulfamethoxypyridazine and sulfathiozole. Two triple drug combinations had the highest activity against amoxicillin-induced round bodies and stationary phase B. burgdorferi persisters: artemisinin/cefoperazone/doxycycline and sulfachlorpyridazine/daptomycin/doxycycline. These findings confirm and extend previous findings that certain drug combinations have superior activity against B. burgdorferi

  6. Freqüência de anticorpos anti-Erhlichia canis, Borrelia burgdorferi e antígenos de Dirofilaria immitis em cães na microrregião Ilhéus-Itabuna, Bahia, Brasil Frequency of antibodies anti-Ehrlichia canis, Borrelia burgdorferi and Dirofilaria immitis antigens in dogs from microrregion Ilhéus-Itabuna, State of Bahia, Brazil

    Directory of Open Access Journals (Sweden)

    Renata S.A. Carlos

    2007-09-01

    Full Text Available Para avaliação de positividade para antígenos de Dirofilaria immitis, anticorpos anti-Borrelia burgdorferi e anti-Ehrlichia canis foram coletadas 200 amostras de sangue de cães, 100 no município de Ilhéus e 100 no de Itabuna. Foi utilizado o "kit" Snap 3DX (IDEXX Laboratories para realização das sorologias. Não se observou nenhum animal positivo para antígenos de D. immitis. Apenas dois dos cães estavam positivos para anticorpos anti-B. burgdorferi. Do total de amostras analisadas, 72 (36% estavam positivas para anticorpos anti-E. canis, sendo 43 em Ilhéus e 29 em Itabuna (p=0,027.In order to detect the positivity to antigens of Dirofilaria immitis, antibodies anti-Borrelia burgdorferi and anti-Ehrlichia canis, 200 canine blood samples were collected as followed: 100 from the municipality of Ilhéus and 100 from Itabuna, State of Bahia. The kit Snap 3DX (IDEXX Laboratories was used to performe serology. None of the tested animals were positive for antigens of D. immitis. Only two dogs of them were positive for antibodies anti-B. burgdorferi. From all the samples analyzed, 72 (36% were positive for antibodies anti-E. canis, 43 from Ilhéus and 29 from Itabuna (p=0,027.

  7. BB0347, from the lyme disease spirochete Borrelia burgdorferi, is surface exposed and interacts with the CS1 heparin-binding domain of human fibronectin.

    Directory of Open Access Journals (Sweden)

    Robert A Gaultney

    Full Text Available The causative agent of Lyme disease, Borrelia burgdorferi, codes for several known fibronectin-binding proteins. Fibronectin a common the target of diverse bacterial pathogens, and has been shown to be essential in allowing for the development of certain disease states. Another borrelial protein, BB0347, has sequence similarity with these other known fibronectin-binding proteins, and may be important in Lyme disease pathogenesis. Herein, we perform an initial characterization of BB0347 via the use of molecular and biochemical techniques. We found that BB0347 is expressed, produced, and presented on the outer surface of intact B. burgdorferi. We also demonstrate that BB0347 has the potential to be important in Lyme disease progression, and have begun to characterize the nature of the interaction between human fibronectin and this bacterial protein. Further work is needed to define the role of this protein in the borrelial infection process.

  8. Prevalence of Babesia canis, Borrelia burgdorferi sensu lato, and Anaplasma phagocytophilum in hard ticks collected from meadows of Lubelskie Voivodship (eastern Poland

    Directory of Open Access Journals (Sweden)

    Dzięgiel Beata

    2014-03-01

    Full Text Available The aim of the study was to assess the distribution of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, and Babesia canis in adult females and males of Ixodes ricinus and Dermacentor reticulatus ticks, inhabiting meadows near large forest complexes throughout the Lubelskie Voivodship (eastern region of Poland. Ticks were collected using the flagging method. Among 720 ticks collected, 506 were identified as D. reticulatus, and 214 as I. ricinus. DNA of B. canis and B. burgdorferi s.l. was detected in 21.3% and 0.6% of D. reticulatus ticks, respectively. In I. ricinus ticks, DNA specific to B. burgdorferi s.l. and A. phagocytophilum was detected in 5.6% and 10.3%, respectively. Co-infections of B. burgdorferi s.l. and A. phagocytophilum were found in two I. ricinus ticks. These results indicate that the Lublin region is an area at risk of tick-borne diseases of humans and animals, which must be considered in clinical practice.

  9. The nucleotide excision repair system of Borrelia burgdorferi is the sole pathway involved in repair of DNA damage by UV light.

    Science.gov (United States)

    Hardy, Pierre-Olivier; Chaconas, George

    2013-05-01

    To survive and avoid accumulation of mutations caused by DNA damage, the genomes of prokaryotes encode a variety of DNA repair pathways most well characterized in Escherichia coli. Some of these are required for the infectivity of various pathogens. In this study, the importance of 25 DNA repair/recombination genes for Borrelia burgdorferi survival to UV-induced DNA damage was assessed. In contrast to E. coli, where 15 of these genes have an effect on survival of UV irradiation, disruption of recombinational repair, transcription-coupled repair, methyl-directed mismatch correction, and repair of arrested replication fork pathways did not decrease survival of B. burgdorferi exposed to UV light. However, the disruption of the B. burgdorferi nucleotide excision repair (NER) pathway (uvrA, uvrB, uvrC, and uvrD) resulted in a 10- to 1,000-fold increase in sensitivity to UV light. A functional NER pathway was also shown to be required for B. burgdorferi resistance to nitrosative damage. Finally, disruption of uvrA, uvrC, and uvrD had only a minor effect upon murine infection by increasing the time required for dissemination. PMID:23475971

  10. BORRELIA BURGDORFERI DNA IN BIOLOGICAL SAMPLES FROM PATIENTS WITH SARCOIDOSIS USING THE POLYMERASE CHAIN REACTION TECHNIQUE

    Institute of Scientific and Technical Information of China (English)

    连伟; 罗慰慈

    1995-01-01

    Polymerase chain reaction (PCR) was used to detect the presence of Borretia burgdoferi DNA in biological samples from patients with sarcoidcsis. The target DNA sequence was of chromosomal origin. The amplified DNA sequence was analyzed by agarose gel electrophoresis, PAGE with silver staining, and the identity of amplified DNA was confirmed by restriction enzyme cleavage and DNA-DNA hybridlzation with a 32P-labelled probe. The assay was sensitive to fewer than two copies of B. burgdor feri genome, even in the presence of a 104-fold excess of human eukaryotic DNA, and was also specific to different B. burgdorferl strains tested. Sera seroiogieally positive to B. burgdorferi (n=26), broncbemlveolar lavage fluid and supematant of BALF (n=26) and peripheral blood (n=9) from sarcoidosis patients were tested. The positive rate was low (4/26, 2/26, and 0/9, respectively). It was considered that DNA from B. bur gdor feri may be identified in a minority of patients with s,arcoidosis, and it may play a pathogenetic rote in such cases. More studies need to be done before advancing the hypothesis of an etiologic role of B. burgdorferi in sarcoidosis.

  11. Isolation of DNA after Extraction of RNA To Detect the Presence of Borrelia burgdorferi and Expression of Host Cellular Genes from the Same Tissue Sample

    Science.gov (United States)

    Amemiya, Kei; Schaefer, Henry; Pachner, Andrew R.

    1999-01-01

    We are investigating the neuropathogenesis of Lyme disease caused by Borrelia burgdorferi in a nonhuman primate model. In the past, two separate pieces of tissue had to be used when both analyzing for the presence of the spirochete and examining the host response to infection. We have modified a procedure to purify DNA from the same sample after the extraction of RNA. The remaining material containing the DNA was precipitated, and residual organic reagent was removed prior to deproteinization and extraction of the DNA. This procedure now allows us to both assay for the presence of the Lyme microorganism and analyze the host response in the same tissue preparation. PMID:10325389

  12. Orale Immunisierung mit rekombinanten "Escherichia coli" NISSLE 1917:Charakterisierung der humoralen Immunantwort gegen Oberflächenexprimierte heterologe Antigene aus "Borrelia burgdorferi" und "Listeria monocytogenes"

    OpenAIRE

    Buddenborg, C. (Christian)

    2005-01-01

    Im Rahmen dieser Arbeit wurde ein Lebendvektor-System zur oralen Immunisierung entwickelt, das aus dem apathogenen, probiotischen E. coli Stamm NISSLE 1917 besteht. Mit Hilfe des AIDA-Autotransporters wurden die äußeren Membranproteine OspA und OspG aus Borrelia burgdorferi bzw. p60 aus Listeria monocytogenes auf der Oberfläche von E. coli NISSLE 1917 präsentiert. Nach oraler Immunisierung der Versuchstiere wurden die spezifischen humoralen Immunantworten gegen die verwendeten Antigene in Ser...

  13. Response to Esteve-Gassent et al.: flaB sequences obtained from Texas PCR products are identical to the positive control strain Borrelia burgdorferi B31

    OpenAIRE

    Norris, Steven J; Barbour, Alan G.; Fish, Durland; Diuk-Wasser, Maria A.

    2015-01-01

    Feria-Arroyo et al. had reported previously that, based on PCR analysis, 45 % of Ixodes scapularis ticks collected in Texas and Mexico were infected with the Lyme disease spirochete Borrelia burgdorferi (Parasit. Vectors 2014, 7:199). However, our analyses of their initial data (Parasit. Vectors 2014, 7:467) and a recent response by Esteve-Gassent et al. (Parasit. Vectors 2015, 8:129) provide evidence that the positive PCR results obtained from both ribosomal RNA intergenic sequences and the ...

  14. Detección de anticuerpos contra Borrelia burgdorferi e identificación de garrapatas ixodidas en Piura Y Amazonas, Perú

    OpenAIRE

    Martha Glenny A; Leonardo Mendoza U; Eduardo Falconí R

    2004-01-01

    Objetivos: Detectar anticuerpos IgG/IgM contra Borrelia burgdorferi en población general, procedentes de los departamentos de Piura y Amazonas e identificar especies de garrapatas probablemente incriminadas en la transmisión de la enfermedad de Lyme. Material y Métodos: Entre agosto del año 2001 y junio de 2002, se colectaron muestras de sangre de 232 pobladores procedentes de ocho localidades del Departamento de Piura y 12 del Departamento de Amazonas, para evaluar mediante ELISA Captia™ Lym...

  15. Detailed analysis of sequence changes occurring during vlsE antigenic variation in the mouse model of Borrelia burgdorferi infection.

    Directory of Open Access Journals (Sweden)

    Loïc Coutte

    2009-02-01

    Full Text Available Lyme disease Borrelia can infect humans and animals for months to years, despite the presence of an active host immune response. The vls antigenic variation system, which expresses the surface-exposed lipoprotein VlsE, plays a major role in B. burgdorferi immune evasion. Gene conversion between vls silent cassettes and the vlsE expression site occurs at high frequency during mammalian infection, resulting in sequence variation in the VlsE product. In this study, we examined vlsE sequence variation in B. burgdorferi B31 during mouse infection by analyzing 1,399 clones isolated from bladder, heart, joint, ear, and skin tissues of mice infected for 4 to 365 days. The median number of codon changes increased progressively in C3H/HeN mice from 4 to 28 days post infection, and no clones retained the parental vlsE sequence at 28 days. In contrast, the decrease in the number of clones with the parental vlsE sequence and the increase in the number of sequence changes occurred more gradually in severe combined immunodeficiency (SCID mice. Clones containing a stop codon were isolated, indicating that continuous expression of full-length VlsE is not required for survival in vivo; also, these clones continued to undergo vlsE recombination. Analysis of clones with apparent single recombination events indicated that recombinations into vlsE are nonselective with regard to the silent cassette utilized, as well as the length and location of the recombination event. Sequence changes as small as one base pair were common. Fifteen percent of recovered vlsE variants contained "template-independent" sequence changes, which clustered in the variable regions of vlsE. We hypothesize that the increased frequency and complexity of vlsE sequence changes observed in clones recovered from immunocompetent mice (as compared with SCID mice is due to rapid clearance of relatively invariant clones by variable region-specific anti-VlsE antibody responses.

  16. Detection of anti-Borrelia burgdorferi antibodies in buffaloes (Bubalus bubalis in the state of Pará, Brazil Detecção de anticorpos anti-Borrelia burgdorferi em búfalos (Bubalus bubalis no estado do Pará, Brasil

    Directory of Open Access Journals (Sweden)

    Fabíola do Nascimento Corrêa

    2012-09-01

    Full Text Available This study aimed to investigate the frequency of homologous antibodies of IgG class against Borrelia burgdorferi in buffaloes in the state of Pará, Brazil. Blood serum samples from 491 buffaloes were analyzed by means of the indirect ELISA test, using crude antigen produced from a cultivar of the North American strain G39/40 of B. burgdorferi. There were 412 positive samples (83.91%, and there was no statistically significant difference in the proportions of positive animals between the 81.69% (232/284 originating from Marajó Island and the 86.96% (180/207 from the continental area of the state of Pará. In all the municipalities studied, the frequency of positive findings of antibodies against B. burgdorferi among the animals ranged from 63.6% to 92.9%. The high numbers of seropositive animals can be explained by the frequent presence of the tick Rhipicephalus (Boophilusmicroplus, and by the possible existence of spirochetes of the genus Borrelia infecting buffaloes in the region studied, although specific studies are needed to confirm this relationship. These factors suggest that a cross-reaction exists between the North American strain G39/40 of B. burgdorferi, which is used as an antigenic substrate, and the species of Borrelia spp. that possibly infects buffaloes in the state of Pará.Este estudo teve como objetivo investigar a frequência de anticorpos homólogos da classe IgG contra Borrelia burgdorferi em búfalos do estado do Pará. Amostras de soro de 491 búfalos foram analisadas por meio do teste ELISA indireto, utilizando antígeno bruto produzido a partir do cultivo da cepa norte americana G39/40 de B. burgdorferi. Foram encontrados 412 soros positivos (83,91%, não havendo diferença estatística significativa entre os 81,69% (232/284 animais positivos provenientes da Ilha de Marajó e os 86,96% (180/207 da base continental do estado do Pará. Em todos os municípios estudados os animais apresentaram frequência de anticorpos

  17. Human TLR8 is activated upon recognition of Borrelia burgdorferi RNA in the phagosome of human monocytes

    Science.gov (United States)

    Cervantes, Jorge L.; La Vake, Carson J.; Weinerman, Bennett; Luu, Stephanie; O'Connell, Caitlin; Verardi, Paulo H.; Salazar, Juan C.

    2013-01-01

    Phagocytosed Borrelia burgdorferi (Bb), the Lyme disease spirochete, induces a robust and complex innate immune response in human monocytes, in which TLR8 cooperates with TLR2 in the induction of NF-κB-mediated cytokine production, whereas TLR8 is solely responsible for transcription of IFN-β through IRF7. We now establish the role of Bb RNA in TLR8-mediated induction of IFN-β. First, using TLR2-transfected HEK.293 cells, which were unable to phagocytose intact Bb, we observed TLR2 activation by lipoprotein-rich borrelial lysates and TLR2 synthetic ligands but not in response to live spirochetes. Purified Bb RNA, but not borrelial DNA, triggered TLR8 activation. Neither of these 2 ligands induced activation of TLR7. Using purified human monocytes we then show that phagocytosed live Bb, as well as equivalent amounts of borrelial RNA delivered into the phagosome by polyethylenimine (PEI), induces transcription of IFN-β and secretion of TNF-α. The cytokine response to purified Bb RNA was markedly impaired in human monocytes naturally deficient in IRAK-4 and in cells with knockdown TLR8 expression by small interfering RNA. Using confocal microscopy we provide evidence that TLR8 colocalizes with internalized Bb RNA in both early (EEA1) and late endosomes (LAMP1). Live bacterial RNA staining indicates that spirochetal RNA does not transfer from the phagosome into the cytosol. Using fluorescent dextran particles we show that phagosomal integrity in Bb-infected monocytes is not affected. We demonstrate, for the first time, that Bb RNA is a TLR8 ligand in human monocytes and that transcription of IFN-β in response to the spirochete is induced from within the phagosomal vacuole through the TLR8-MyD88 pathway. PMID:23906644

  18. Use of nonelectrolytes reveals the channel size and oligomeric constitution of the Borrelia burgdorferi P66 porin.

    Directory of Open Access Journals (Sweden)

    Iván Bárcena-Uribarri

    Full Text Available In the Lyme disease spirochete Borrelia burgdorferi, the outer membrane protein P66 is capable of pore formation with an atypical high single-channel conductance of 11 nS in 1 M KCl, which suggested that it could have a larger diameter than 'normal' Gram-negative bacterial porins. We studied the diameter of the P66 channel by analyzing its single-channel conductance in black lipid bilayers in the presence of different nonelectrolytes with known hydrodynamic radii. We calculated the filling of the channel with these nonelectrolytes and the results suggested that nonelectrolytes (NEs with hydrodynamic radii of 0.34 nm or smaller pass through the pore, whereas neutral molecules with greater radii only partially filled the channel or were not able to enter it at all. The diameter of the entrance of the P66 channel was determined to be ≤1.9 nm and the channel has a central constriction of about 0.8 nm. The size of the channel appeared to be symmetrical as judged from one-sidedness of addition of NEs. Furthermore, the P66-induced membrane conductance could be blocked by 80-90% by the addition of the nonelectrolytes PEG 400, PEG 600 and maltohexaose to the aqueous phase in the low millimolar range. The analysis of the power density spectra of ion current through P66 after blockage with these NEs revealed no chemical reaction responsible for channel block. Interestingly, the blockage of the single-channel conductance of P66 by these NEs occurred in about eight subconductance states, indicating that the P66 channel could be an oligomer of about eight individual channels. The organization of P66 as a possible octamer was confirmed by Blue Native PAGE and immunoblot analysis, which both demonstrated that P66 forms a complex with a mass of approximately 460 kDa. Two dimension SDS PAGE revealed that P66 is the only polypeptide in the complex.

  19. Assessment of decorin-binding protein A to the infectivity of Borrelia burgdorferi in the murine models of needle and tick infection

    Directory of Open Access Journals (Sweden)

    Hagman Kayla E

    2008-05-01

    Full Text Available Abstract Background Decorin-binding proteins (Dbps A and B of Borrelia burgdorferi, the agent of Lyme disease, are surface-exposed lipoproteins that presumably bind to the extracellular matrix proteoglycan, decorin. B. burgdorferi infects various tissues including the bladder, heart, joints, skin and the central nervous system, and the ability of B. burgdorferi to bind decorin has been hypothesized to be important for this disseminatory pathogenic strategy. Results To determine the role of DbpBA in the infectious lifecycle of B. burgdorferi, we created a DbpBA-deficient mutant of B. burgdorferi strain 297 and compared the infectious phenotype of the mutant to the wild-type strain in the experimental murine model of Lyme borreliosis. The mutant strain exhibited a 4-log decrease in infectivity, relative to the wild-type strain, when needle inoculated into mice. Upon complementation of the DbpBA-mutant strain with DbpA, the wild-type level of infectivity was restored. In addition, we demonstrated that the DbpBA-deficient mutant was able to colonize Ixodes scapularis larval ticks after feeding on infected mice and persist within the ticks during the molt to the nymphal state. Moreover, surprisingly, the DbpBA-mutant strain was capable of being transmitted to naïve mice via tick bite, giving rise to infected mice. Conclusion These results suggest that DbpBA is not required for the natural tick-transmission process to mammals, despite inferences from needle-inoculation experiments implying a requirement for DbpBA during mammalian infection. The combined findings also send a cautionary note regarding how results from needle-inoculation experiments with mice should be interpreted.

  20. Use of T7 RNA polymerase to direct expression of outer Surface Protein A (OspA) from the Lyme disease Spirochete, Borrelia burgdorferi

    Science.gov (United States)

    Dunn, John J.; Lade, Barbara N.

    1991-01-01

    The OspA gene from a North American strain of the Lyme disease Spirochete, Borrelia burgdorferi, was cloned under the control of transciption and translation signals from bacteriophage T7. Full-length OspA protein, a 273 amino acid (31kD) lipoprotein, is expressed poorly in Escherichia coli and is associated with the insoluble membrane fraction. In contrast, a truncated form of OspA lacking the amino-terminal signal sequence which normally would direct localization of the protein to the outer membrane is expressed at very high levels (less than or equal to 100 mg/liter) and is soluble. The truncated protein was purified to homogeneity and is being tested to see if it will be useful as an immunogen in a vaccine against Lyme disease. Circular dichroism and fluorescence spectroscopy was used to characterize the secondary structure and study conformational changes in the protein. Studies underway with other surface proteins from B burgdorferi and a related spirochete, B. hermsii, which causes relapsing fever, leads us to conclude that a strategy similar to that used to express the truncated OspA can provide a facile method for producing variations of Borrelia lipoproteins which are highly expressed in E. coli and soluble without exposure to detergents.

  1. Study of a Cohort of 1,886 Persons To Determine Changes in Antibody Reactivity to Borrelia burgdorferi 3 Months after a Tick Bite

    DEFF Research Database (Denmark)

    Dessau, Ram B; Fryland, Linda; Wilhelmsson, Peter;

    2015-01-01

    Lyme borreliosis is a tick-borne disease caused by the bacterium Borrelia burgdorferi. The most frequent clinical manifestation is a rash called erythema migrans. Changes in antibody reactivity to B. burgdorferi 3 months after a tick bite are measured using enzyme-linked immunosorbent assays...... (ELISAs). One assay is based on native purified flagellum antigen (IgG), and the other assay is based on a recombinant antigen called C6 (IgG or IgM). Paired samples were taken at the time of a tick bite and 3 months later from 1,886 persons in Sweden and the Åland Islands, Finland. The seroconversion or...... antigen and the C6 antigen, respectively. Graphical methods to display the antibody response and to choose thresholds for a rise in relative antibody reactivity are shown and discussed. In conclusion, 5.4% of people with tick bites showed a rise in Borrelia-specific antibodies above the 2.5% threshold in...

  2. Two Different Virulence-Related Regulatory Pathways in Borrelia burgdorferi Are Directly Affected by Osmotic Fluxes in the Blood Meal of Feeding Ixodes Ticks.

    Science.gov (United States)

    Bontemps-Gallo, Sébastien; Lawrence, Kevin; Gherardini, Frank C

    2016-08-01

    Lyme disease, caused by Borrelia burgdorferi, is a vector-borne illness that requires the bacteria to adapt to distinctly different environments in its tick vector and various mammalian hosts. Effective colonization (acquisition phase) of a tick requires the bacteria to adapt to tick midgut physiology. Successful transmission (transmission phase) to a mammal requires the bacteria to sense and respond to the midgut environmental cues and up-regulate key virulence factors before transmission to a new host. Data presented here suggest that one environmental signal that appears to affect both phases of the infective cycle is osmolarity. While constant in the blood, interstitial fluid and tissue of a mammalian host (300 mOsm), osmolarity fluctuates in the midgut of feeding Ixodes scapularis. Measured osmolarity of the blood meal isolated from the midgut of a feeding tick fluctuates from an initial osmolarity of 600 mOsm to blood-like osmolarity of 300 mOsm. After feeding, the midgut osmolarity rebounded to 600 mOsm. Remarkably, these changes affect the two independent regulatory networks that promote acquisition (Hk1-Rrp1) and transmission (Rrp2-RpoN-RpoS) of B. burgdorferi. Increased osmolarity affected morphology and motility of wild-type strains, and lysed Hk1 and Rrp1 mutant strains. At low osmolarity, Borrelia cells express increased levels of RpoN-RpoS-dependent virulence factors (OspC, DbpA) required for the mammalian infection. Our results strongly suggest that osmolarity is an important part of the recognized signals that allow the bacteria to adjust gene expression during the acquisition and transmission phases of the infective cycle of B. burgdorferi. PMID:27525653

  3. Co-infection of blacklegged ticks with Babesia microti and Borrelia burgdorferi is higher than expected and acquired from small mammal hosts.

    Directory of Open Access Journals (Sweden)

    Michelle H Hersh

    Full Text Available Humans in the northeastern and midwestern United States are at increasing risk of acquiring tickborne diseases--not only Lyme disease, but also two emerging diseases, human granulocytic anaplasmosis and human babesiosis. Co-infection with two or more of these pathogens can increase the severity of health impacts. The risk of co-infection is intensified by the ecology of these three diseases because all three pathogens (Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti are transmitted by the same vector, blacklegged ticks (Ixodes scapularis, and are carried by many of the same reservoir hosts. The risk of exposure to multiple pathogens from a single tick bite and the sources of co-infected ticks are not well understood. In this study, we quantify the risk of co-infection by measuring infection prevalence in 4,368 questing nymphs throughout an endemic region for all three diseases (Dutchess County, NY to determine if co-infections occur at frequencies other than predicted by independent assortment of pathogens. Further, we identify sources of co-infection by quantifying rates of co-infection on 3,275 larval ticks fed on known hosts. We find significant deviations of levels of co-infection in questing nymphs, most notably 83% more co-infection with Babesia microti and Borrelia burgdorferi than predicted by chance alone. Further, this pattern of increased co-infection was observed in larval ticks that fed on small mammal hosts, but not on meso-mammal, sciurid, or avian hosts. Co-infections involving A. phagocytophilum were less common, and fewer co-infections of A. phagocytophilum and B. microti than predicted by chance were observed in both questing nymphs and larvae fed on small mammals. Medical practitioners should be aware of the elevated risk of B. microti/B. burgdorferi co-infection.

  4. Mechanism of infection transmission and pathogenesis of Borrelia burgdorferi%莱姆病螺旋体的转移感染和致病机制

    Institute of Scientific and Technical Information of China (English)

    郝琴; 万康林; 徐建国

    2011-01-01

    本文主要就莱姆病螺旋体从蜱到宿主动物的转移感染以及在宿主体内存活并致病的机制进行了的阐述,包括螺旋体在蜱内定居以及感染宿主动物过程中蛋白的变化及作用;螺旋体在宿主体内致病的各种机制(病原体本身的作用、免疫逃避、细胞因子的作用、自身免疫).%The paper describes the mechanism of transmission of Borrelia burgdorferi from ticks to host animals and pathogenesis in host animals, including the colonizing of B. Burgdorferi in ticks, the change and role of its proteins in infecting host animals and all the pathogenic mechanisms of B. Burgdorferi in host animals (self role of pathogen, immune evasion, function of cytokines and autoimmunity).

  5. Immunohistochemistry and real-time PCR as diagnostic tools for detection of Borrelia burgdorferi sensu lato in ticks collected from humans.

    Science.gov (United States)

    Briciu, Violeta T; Sebah, Daniela; Coroiu, Georgiana; Lupşe, Mihaela; Cârstina, Dumitru; Ţăţulescu, Doina F; Mihalca, Andrei D; Gherman, Călin M; Leucuţa, Daniel; Meyer, Fabian; Hizo-Teufel, Cecilia; Fingerle, Volker; Huber, Ingrid

    2016-05-01

    The objective of this study was to evaluate different methods used for detection of Borrelia burgdorferi sensu lato (s.l.) in ticks: immunohistochemistry followed by focus floating microscopy (FFM) and real-time polymerase chain reaction (real-time PCR) targeting the ospA and hbb genes. Additionally, an optimized ospA real-time PCR assay was developed with an integrated internal amplification control (IAC) for the detection of inhibition in the PCR assay and was validated as an improved screening tool for B. burgdorferi. One hundred and thirty-six ticks collected from humans in a hospital from Cluj-Napoca, Romania, were investigated regarding genus, stage of development and sex, and then tested by all three assays. A poor quality of agreement was found between FFM and each of the two real-time PCR assays, as assessed by concordance analysis (Cohen's kappa), whereas the agreement between the two real-time PCR assays was moderate. The present study argues for a low sensitivity of FFM and underlines that discordant results of different assays used for detection of B. burgdorferi in ticks are frequent. PMID:26801157

  6. Eradication of Biofilm-like Microcolony Structures of Borrelia burgdorferi by Daunomycin and Daptomycin but not Mitomycin C in Combination with Doxycycline and Cefuroxime

    Directory of Open Access Journals (Sweden)

    Jie eFeng

    2016-02-01

    Full Text Available Lyme disease, caused by Borrelia burgdorferi, is the most common vector-borne disease in the United States and Europe. While the majority of Lyme disease patients can resolve their symptoms if treated promptly, 10-20% of patients suffer from prolonged symptoms called post-treatment Lyme disease syndrome (PTLDS. Although the cause for PTLDS is unclear, one possibility is the presence of bacterial persisters not effectively cleared by the current Lyme antibiotics. Recent studies identified several drug candidates including daptomycin, daunomycin, doxorubicin, and mitomycin C that had good activity against B. burgdorferi persisters. However, their relative activities against B. burgdorferi persisters have not been evaluated under the same conditions. In this study, we tested the anti-persister activities of these drugs against both 7-day and 15-day old stationary phase cultures of B. burgdorferi individually as well as in combination with Lyme antibiotics doxycycline and cefuroxime (Ceftin. Our findings demonstrate daunomycin and daptomycin were more active than mitomycin C in single drug comparison at 10 and 20 µM, as well as in drug combinations with doxycycline and cefuroxime. In addition, daunomycin was more active than doxorubicin which correlated with their ability to stain and accumulate in B. burgdorferi. The two drug combination of doxycycline and cefuroxime was unable to eradicate biofilm-like microcolonies of B. burgdorferi persisters. However, the addition of either daunomycin or daptomycin to the doxycycline + cefuroxime combination completely eradicated the biofilm-like structures and produced no visible bacterial regrowth after 7 days and 21 days, while the addition of doxorubicin was unable to prevent regrowth at either 7 day or 21 day subculture. Mitomycin C in combination with doxycycline and cefuroxime caused no regrowth at 7 days but visible spirochetal regrowth occurred after 21 day subculture. Furthermore, we found that

  7. NMR studies of Borrelia burgdorferi OspA, a 28 kDa protein containing a single-layer {beta}-sheet

    Energy Technology Data Exchange (ETDEWEB)

    Pham, Thuy-Nga; Koide, Shohei

    1998-05-15

    The crystal structure of outer surface protein A (OspA) from Borrelia burgdorferi contains a single-layer {beta}-sheet connecting the N- and C-terminal globular domains. The central {beta}-sheet consists largely of polar amino acids and it is solvent-exposed on both faces, which so far appears to be unique among known protein structures. We have accomplished nearly complete backbone H, C and N and C{sup ;}/H{sup {beta}} assignments of OspA (28 kDa) using standard triple resonance techniques without perdeuteration. This was made possible by recording spectra at a high temperature (45 {sup o}C ). The chemical shift index and {sup 15}N T{sub 1}/T{sub 2} ratios show that both the secondary structure and the global conformation of OspA in solution are similar to the crystal structure, suggesting that the unique central {beta}-sheet is fairly rigid.

  8. Borrelia burgdorferi Requires the Alternative Sigma Factor RpoS for Dissemination within the Vector during Tick-to-Mammal Transmission

    Science.gov (United States)

    Dunham-Ems, Star M.; Caimano, Melissa J.; Eggers, Christian H.; Radolf, Justin D.

    2012-01-01

    While the roles of rpoSBb and RpoS-dependent genes have been studied extensively within the mammal, the contribution of the RpoS regulon to the tick-phase of the Borrelia burgdorferi enzootic cycle has not been examined. Herein, we demonstrate that RpoS-dependent gene expression is prerequisite for the transmission of spirochetes by feeding nymphs. RpoS-deficient organisms are confined to the midgut lumen where they transform into an unusual morphotype (round bodies) during the later stages of the blood meal. We show that round body formation is rapidly reversible, and in vitro appears to be attributable, in part, to reduced levels of Coenzyme A disulfide reductase, which among other functions, provides NAD+ for glycolysis. Our data suggest that spirochetes default to an RpoS-independent program for round body formation upon sensing that the energetics for transmission are unfavorable. PMID:22359504

  9. Biochemical and biophysical characterization of the major outer surface protein, OSP-A from North American and European isolates of Borrelia burgdorferi

    Energy Technology Data Exchange (ETDEWEB)

    McGrath, B.C.; Dunn, J.J. [Brookhaven National Lab., Upton, NY (United States); France, L.L. [Plum Island Animal Disease Center, Greenport, NY (United States); Jaing, W.; Polin, D.; Gorgone, G.; Luft, B.; Dykhuizen, D. [SUNY at Stony Brook, Stony Brook, NY (United States)

    1995-12-31

    Lyme borreliosis, caused by the spirochete Borrelia burgdorferi, is the most common vector-borne disease in North America and Western Europe. As the major delayed immune response in humans, a better understanding of the major outer surface lipoproteins OspA and OspB are of much interest. These proteins have been shown to exhibit three distinct phylogenetic genotypes based on their DNA sequences. This paper describes the cloning of genomic DNA for each variant and amplification of PCR. DNA sequence data was used to derive computer driven phylogenetic analysis and deduced amino acid sequences. Overproduction of variant OspAs was carried out in E. coli using a T7-based expression system. Circular dichroism and fluorescence studies was carried out on the recombinant B31 PspA yielding evidence supporting a B31 protein containing 11% alpha-helix, 34% antiparallel beta-sheet, 12% parallel beta sheet.

  10. 广西首次分离到莱姆病螺旋体%Borrelia burgdorferi isolated from rat in Yachang forest farm of Guangxi Province

    Institute of Scientific and Technical Information of China (English)

    刘敏; 王树声; 张涛; 曾霞; 杨光华; 王昭孝

    2001-01-01

    Objective To isolate Lyme disease spirochetes from it′ s host-rat in Yachang forest farm of Guangxi Province.Methods The rat's tissues of urinary bladder were inoculated into the BSK-Ⅱ medium.After incubating for 7~ 14 days,the Borrelia burgdorferi was observed in dark-field microscope.Results From these rats,six isolates were obtained.They were named GXLD4,8,9,16,18,20.The positive rate of isolation is 40% (6/15).Conclusion It is the first report that Borrelia burgdorferi was isolated from Guangxi Province.%目的从广西鼠类的膀胱中分离莱姆病螺旋体。方法膀胱组织标本接种 BSK-Ⅱ培养基进行莱姆病病原分离培养。结果从广西雅长林场捕获的 23只野鼠( 8只鼠标本培养污染)中共分离出 6株莱姆病螺旋体,分别命名为 GXLD- 4,8,9,16,18,20,分离阳性率为 40% (6/15)。结论首次从广西莱姆病宿主动物中分离出伯氏螺旋体,证实广西存在莱姆病的流行。

  11. Serological survey of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, and Ehrlichia canis infections in rural and urban dogs in Central Italy

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    Valentina Virginia Ebani

    2014-11-01

    Full Text Available [b]Introduction[i][/i][/b][i]. Borrelia burgdorferi [/i]sensu lato (s.l. and Anaplasma phagocytophilum are well known zoonotic pathogens, whereas[i] Ehrlichia canis[/i] is usually considered to be of veterinary concern, although on the basis of recent reports it also seems to be able to infect humans. [b]objective[/b]. The aim of the study was to determine the seroprevalence of [i]B. burgdorferi [/i]s.l., A. phagocytophilum and [i]E. canis[/i] in an Italian canine population, and to verify if there are differences between dogs living in urban areas and those from a rural environment. [b]materials and method.[/b] Blood sera of 1,965 dogs, 1,235 from cities and 730 from rural areas, were tested by indirect immunofluorescent assay (IFAT. [b]results[/b]. The overall seroprevalence was highest for E. canis (7.07%, followed by [i]A. phagocytophilum[/i] (4.68%, and [i]B. burgdorferi[/i] s.l. (1.47%. Rural dogs showed the highest seroprevalence to [i]B. burgdorferi[/i] s.l. and [i]A. phagocytophilum[/i]. No significant differences were observed between rural and urban [i]E. canis[/i]-positive dogs. A low percentage (1.32% of dogs with dual seropositivity was detected, and no triple positive reactions were observed. No significant differences were detected in the seroprevalence of the three agents in relationship to the age and gender of the dogs. Seroprevalence in the five years considered were not statistically different, except for the lowest rate for [i]E. canis[/i] observed in 2012. [b]conclusions[/b]. The results confirm the presence of [i]B. burgdorferi[/i] s.l., [i]A. phagocytophilum[/i] and [i]E. canis[/i] in Italian dogs in both urban and rural areas. Monitoring pet dogs, which share the same environment with their owners, is useful for identifying the presence of tick-borne disease agents of both veterinary and public health significance

  12. Investigation of the genes involved in antigenic switching at the vlsE locus in Borrelia burgdorferi: an essential role for the RuvAB branch migrase.

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    Ashley R Dresser

    2009-12-01

    Full Text Available Persistent infection by pathogenic organisms requires effective strategies for the defense of these organisms against the host immune response. A common strategy employed by many pathogens to escape immune recognition and clearance is to continually vary surface epitopes through recombinational shuffling of genetic information. Borrelia burgdorferi, a causative agent of Lyme borreliosis, encodes a surface-bound lipoprotein, VlsE. This protein is encoded by the vlsE locus carried at the right end of the linear plasmid lp28-1. Adjacent to the expression locus are 15 silent cassettes carrying information that is moved into the vlsE locus through segmental gene conversion events. The protein players and molecular mechanism of recombinational switching at vlsE have not been characterized. In this study, we analyzed the effect of the independent disruption of 17 genes that encode factors involved in DNA recombination, repair or replication on recombinational switching at the vlsE locus during murine infection. In Neisseria gonorrhoeae, 10 such genes have been implicated in recombinational switching at the pilE locus. Eight of these genes, including recA, are either absent from B. burgdorferi, or do not show an obvious requirement for switching at vlsE. The only genes that are required in both organisms are ruvA and ruvB, which encode subunits of a Holliday junction branch migrase. Disruption of these genes results in a dramatic decrease in vlsE recombination with a phenotype similar to that observed for lp28-1 or vls-minus spirochetes: productive infection at week 1 with clearance by day 21. In SCID mice, the persistence defect observed with ruvA and ruvB mutants was fully rescued as previously observed for vlsE-deficient B. burgdorferi. We report the requirement of the RuvAB branch migrase in recombinational switching at vlsE, the first essential factor to be identified in this process. These findings are supported by the independent work of Lin et

  13. Methylated DNA in Borrelia species.

    OpenAIRE

    Hughes, C A; Johnson, R C

    1990-01-01

    The DNA of Borrelia species was examined for the presence of methylated GATC sequences. The relapsing-fever Borrelia sp., B. coriaceae, and only 3 of 22 strains of B. burgdorferi contained adenine methylation systems. B. anserina lacked an adenine methylation system. Fundamental differences in DNA methylation exist among members of the genus Borrelia.

  14. Evaluation of antibodies production against Borrelia burgdorferi in cattle submitted to rBm86 protein Boophilus microplus tick immunization and associated challenges influence Avaliação da produção de anticorpos anti Borrelia burgdorferi em bovinos submetidos à imunização com proteína rBm86 de carrapato Boophilus microplus e influência dos desafios associados

    OpenAIRE

    Márcia Mayumi Ishikawa; Adivaldo Henrique Fonseca; Natalino Hajime Yoshinari; Ana Luiza Alves Rosa Osório

    2003-01-01

    IgG antibodies production against Borrelia burgdorferi in immunized cattle with rBm86 protein from Boophilus microplus was evaluated as well as the influence of the association between immunizations and stress through indirect ELISA test during one year. In the present study there was no influence of the isolated challenged used on the production of IgG antibodies against B. burgdorferi. The rBm86 immunogen did not cause significant oscillation in the production of IgG antibodies against B. b...

  15. ErpC, a member of the complement regulator-acquiring family of surface proteins from Borrelia burgdorferi, possesses an architecture previously unseen in this protein family

    International Nuclear Information System (INIS)

    The structure of ErpC, a member of the complement regulator-acquiring surface protein family from B. burgdorferi, has been solved, providing insights into the strategies of complement evasion by this zoonotic bacterium and suggesting a common architecture for other members of this protein family. Borrelia burgdorferi is a spirochete responsible for Lyme disease, the most commonly occurring vector-borne disease in Europe and North America. The bacterium utilizes a set of proteins, termed complement regulator-acquiring surface proteins (CRASPs), to aid evasion of the human complement system by recruiting and presenting complement regulator factor H on its surface in a manner that mimics host cells. Presented here is the atomic resolution structure of a member of this protein family, ErpC. The structure provides new insights into the mechanism of recruitment of factor H and other factor H-related proteins by acting as a molecular mimic of host glycosaminoglycans. It also describes the architecture of other CRASP proteins belonging to the OspE/F-related paralogous protein family and suggests that they have evolved to bind specific complement proteins, aiding survival of the bacterium in different hosts

  16. Further structural insights into the binding of complement factor H by complement regulator-acquiring surface protein 1 (CspA) of Borrelia burgdorferi

    International Nuclear Information System (INIS)

    B. burgdorferi binds complement factor H using a dimeric surface protein, CspA (BbCRASP-1). Presented here is a new structure of CspA that suggests that there is a degree of flexibility between subunits which may have implications for complement regulator binding. Borrelia burgdorferi has evolved many mechanisms of evading the different immune systems across its range of reservoir hosts, including the capture and presentation of host complement regulators factor H and factor H-like protein-1 (FHL-1). Acquisition is mediated by a family of complement regulator-acquiring surface proteins (CRASPs), of which the atomic structure of CspA (BbCRASP-1) is known and shows the formation of a homodimeric species which is required for binding. Mutagenesis studies have mapped a putative factor H binding site to a cleft between the two subunits. Presented here is a new atomic structure of CspA which shows a degree of flexibility between the subunits which may be critical for factor H scavenging by increasing access to the binding interface and allows the possibility that the assembly can clamp around the bound complement regulators

  17. Retrospective analysis of clinical and laboratory findings in hunting dogs with serologic reactions to tick-borne pathogens (Anaplasma phagocytophilum, Borrelia burgdorferi, Babesia canis, Ehrlichia canis, Ricketsia conorii

    Directory of Open Access Journals (Sweden)

    Spasojević-Kosić Ljubica

    2015-01-01

    Full Text Available Seroprevalence of tick-borne infections in endemic areas could be high. In this study, we investigated the seroprevalence of tick-borne pathogens (Anaplasma phagocytophilum, Borrelia burgdorferi, Babesia canis, Ehrlichia canis, Rickettsia conorii in hunting dogs, naturally infected with one or more pathogens. Serological test results of the investigated animals were compared to those from clinical examination, as well as from haematological and biochemical analyses. A total of 74.14% dogs were seropositive (R.conorii 44.83%, B. canis 32.76%, B. burgdorferi 25.86%, E. canis 13.79%, A. phagocytophilum 8.47%, with 25.86% of dogs seropositive to two pathogens, 15.52% seropositive to three pathogens, and 1.72% of dogs seropositive to four pathogens. Among all registered clinical signs, only pyrexia (p<0.05 and arrhythmia (p<0.05 were significant in seropositive dogs. There was no significant difference between seropositive and seronegative dogs regarding the majority of haematological and biochemical parameters. Statistically significant difference was registered for particular haematological (number of red blood cells and seroreactivity to B. burgdorferi and biochemical parameters (albumin concentration and seroreactivity to E. canis, and AST and seroreactivity to R. conorii but these values were not clinically significant. The high exposure to tick-borne pathogens suggests that ectoparasitic profilactic treatment is not adequate in examined population of hunting dogs. Clinical finding of pyrexia need to be further investigated and explained etiologically, which means that molecular diagnosis should be used in order to identify larger number of pathogens because of the possibility of coinfection. [Projekat Ministarstva nauke Republike Srbije, br. TR 31084

  18. The Borrelia burgdorferi telomere resolvase, ResT, anneals ssDNA complexed with its cognate ssDNA-binding protein.

    Science.gov (United States)

    Huang, Shu Hui; Kobryn, Kerri

    2016-06-20

    Spirochetes of the genus Borrelia possess unusual genomes that consist in a linear chromosome and multiple linear and circular plasmids. The linear replicons are terminated by covalently closed hairpin ends, referred to as hairpin telomeres. The hairpin telomeres represent a simple solution to the end-replication problem. Deoxyribonucleic acid replication initiates internally and proceeds bidirectionally toward the hairpin telomeres. The telomere resolvase, ResT, forms the hairpin telomeres from replicated telomere intermediates in a reaction with similarities to those promoted by type IB topoisomerases and tyrosine recombinases. ResT has also been shown to possess DNA single-strand annealing activity. We report here that ResT promotes single-strand annealing of both free DNA strands and ssDNA complexed with single-stranded DNA binding protein (SSB). The annealing of complementary strands bound by SSB requires a ResT-SSB interaction that is mediated by the conserved amphipathic C-terminal tail of SSB. These properties of ResT are similar to those demonstrated for the recombination mediator protein, RecO, of the RecF pathway. Borrelia burgdorferi is unusual in lacking identifiable homologs of the RecFOR proteins. We propose that ResT may provide missing RecFOR functions. PMID:27131360

  19. Prevalence of Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in apparently healthy and CVBD-suspect dogs in Portugal - a national serological study

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    Cardoso Luís

    2012-03-01

    Full Text Available Abstract Background Canine vector-borne diseases (CVBDs are caused by a wide range of pathogens transmitted to dogs by arthropods including ticks and insects. Many CVBD-agents are of zoonotic concern, with dogs potentially serving as reservoirs and sentinels for human infections. The present study aimed at assessing the seroprevalence of infection with or exposure to Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in dogs in Portugal. Methods Based on 120 veterinary medical centres from all the regions of mainland and insular Portugal, 557 apparently healthy and 628 CVBD-suspect dogs were sampled. Serum, plasma or whole blood was tested for qualitative detection of D. immitis antigen and antibodies to E. canis, B. burgdorferi s. l., Anaplasma spp. and L. infantum with two commercial in-clinic enzyme-linked immunosorbent assay kits. Odds ratios (OR were calculated by logistic regression analysis to identify independent risk factors of exposure to the vector-borne agents. Results Total positivity levels to D. immitis, E. canis, B. burgdorferi, Anaplasma spp., L. infantum, one or more agents and mixed agents were 3.6%, 4.1%, 0.2%, 4.5%, 4.3%, 14.0% and 2.0% in the healthy group, and 8.9%, 16.4%, 0.5%, 9.2%, 25.2%, 46.3% and 11.6% in the clinically suspect group, respectively. Non-use of ectoparasiticides was a risk factor for positivity to one or more agents both in the apparently healthy (OR = 2.1 and CVBD-suspect (OR = 1.5 dogs. Seropositivity to L. infantum (OR = 7.6, E. canis (OR = 4.1 and D. immitis (OR = 2.4 were identified as risk factors for the presence of clinical signs compatible with CVBDs. Positivity to mixed agents was not found to be a risk factor for disease. Conclusions Dogs in Portugal are at risk of becoming infected with vector-borne pathogens, some of which are of zoonotic concern. CVBDs should be considered by practitioners and prophylactic measures must be put in

  20. Anti-inflammatory effects of dexamethasone and meloxicam on Borrelia burgdorferi-induced inflammation in neuronal cultures of dorsal root ganglia and myelinating cells of the peripheral nervous system

    OpenAIRE

    Ramesh, Geeta; Meisner, Olivia C.; Philipp, Mario T.

    2015-01-01

    Background Lyme neuroborreliosis (LNB), caused by the spirochete Borrelia burgdorferi (Bb), could result in cognitive impairment, motor dysfunction, and radiculoneuritis. We hypothesized that inflammation is a key factor in LNB pathogenesis and recently evaluated the effects of dexamethasone, a steroidal anti-inflammatory drug, and meloxicam a non-steroidal anti-inflammatory drug (NSAID), in a rhesus monkey model of acute LNB. Dexamethasone treatment significantly reduced the levels of immune...

  1. A possible role for inflammation in mediating apoptosis of oligodendrocytes as induced by the Lyme disease spirochete Borrelia burgdorferi

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    Ramesh Geeta

    2012-04-01

    Full Text Available Abstract Background Inflammation caused by the Lyme disease spirochete B. burgdorferi is an important factor in the pathogenesis of Lyme neuroborreliosis. Our central hypothesis is that B. burgdorferi can cause disease via the induction of inflammatory mediators such as cytokines and chemokines in glial and neuronal cells. Earlier we demonstrated that interaction of B. burgdorferi with brain parenchyma induces inflammatory mediators in glial cells as well as glial (oligodendrocyte and neuronal apoptosis using ex vivo and in vivo models of experimentation. Methods In this study we evaluated the ability of live B. burgdorferi to elicit inflammation in vitro in differentiated human MO3.13 oligodendrocytes and in differentiated primary human oligodendrocytes, by measuring the concentration of immune mediators in culture supernatants using Multiplex ELISA assays. Concomitant apoptosis was quantified in these cultures by the in situ terminal deoxynucleotidyl transferase mediated UTP nick end labeling (TUNEL assay and by quantifying active caspase-3 by flow cytometry. The above phenomena were also evaluated after 48 h of stimulation with B. burgdorferi in the presence and absence of various concentrations of the anti-inflammatory drug dexamethasone. Results B. burgdorferi induced enhanced levels of the cytokine IL-6 and the chemokines IL-8 and CCL2 in MO3.13 cells as compared to basal levels, and IL-8 and CCL2 in primary human oligodendrocytes, in a dose-dependent manner. These cultures also showed significantly elevated levels of apoptosis when compared with medium controls. Dexamethasone reduced both the levels of immune mediators and apoptosis, also in a manner that was dose dependent. Conclusions This finding supports our hypothesis that the inflammatory response elicited by the Lyme disease spirochete in glial cells contributes to neural cell damage. As oligodendrocytes are vital for the functioning and survival of neurons, the inflammation and

  2. Exposure to ticks and seroprevalence of [i]Borrelia burgdorferi [/i]among a healthy young population living in the area of southern Podlasie, Poland

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    Anna Pańczuk

    2014-09-01

    Full Text Available [b]Objectives[/b]. The objective of the study was assessment of risk of infection with [i]Borrelia burgdorferi[/i] in the area of southern Podlasie in Poland, near the border with Belarus, by analysis of post-exposure procedure, and evaluation of asymptomatic infection in adolescents bitten by a tick, confirmed by serologic tests. [b]Material and methods[/b]. The study was conducted among 128 healthy individuals aged 16–20 who declared being bitten by a tick. The level of IgM and IgG class antibodies was determined using the immunoenzymatic test (Borrelia 14 kD + OspC IgM ELISA and Borrelia IgG + VlsE ELISA, DRG Diagnostics. Positive and doubtful results were confirmed using the Western blot method (EUROLINE-WB, EUROIMMUN. [b]Results[/b]. In the study group, the largest number of respondents (59.4% declared tick bite in the region of the lower extremities, most often in the knee pit. Among the methods for removing the tick the largest number of respondents indicated removing it with the use of tweezers, with a simple, swift steady movement (29.7%, and pulling it out with the fingers (22.7%. In the ELISA test, a positive or doubtful result in at least one class was observed in 25.0% of respondents (n=32/128: in IgM class – 23.4% (n=30/128, and in IgG class – 4.7% (n=6/128. After verification with the Western blot test, infection was confirmed in 5.5% of respondents (n=7/128: in IgM class – 1.6% (n=2/128, in IgG class – 3.9% (n=5/128. In IgM class antibodies, the Western blot test confirmed positive or doubtful results of the ELISA test in 6.7%, while in IgG class antibodies in 83.3%. [b]Conclusion[/b]. Evaluation of the actual infection with [i]Borrelia spp.[/i] using serologic tests is difficult due to a certain non-specificity of the ELISA test, especially in IgM class antibodies, and difficulties with performance of a wide scope of specific Western blot tests. The variety of methods of tick removal declared by adolescents suggests

  3. Evaluation of antibodies production against Borrelia burgdorferi in cattle submitted to rBm86 protein Boophilus microplus tick immunization and associated challenges influence Avaliação da produção de anticorpos anti Borrelia burgdorferi em bovinos submetidos à imunização com proteína rBm86 de carrapato Boophilus microplus e influência dos desafios associados

    Directory of Open Access Journals (Sweden)

    Márcia Mayumi Ishikawa

    2003-11-01

    Full Text Available IgG antibodies production against Borrelia burgdorferi in immunized cattle with rBm86 protein from Boophilus microplus was evaluated as well as the influence of the association between immunizations and stress through indirect ELISA test during one year. In the present study there was no influence of the isolated challenged used on the production of IgG antibodies against B. burgdorferi. The rBm86 immunogen did not cause significant oscillation in the production of IgG antibodies against B. burgdorferi capable to interfere in the serological results for Lyme Borreliosis in cattle. This study demonstrated the possibility of transitory changes in the production of antibodies after the association of vaccine stimuli and stress, emphasizing the necessity of serological studies combined with epidemiological and management data.Avaliou-se a produção de anticorpos da classe Ig-G anti Borrelia burgdorferi em bovinos imunizados com proteína recombinante Bm86 de Boophilus microplus, assim como a influência de imunizações e estresse associados por meio do teste ELISA indireto no período de um ano. Não houve interferência na produção de anticorpos IgG anti B. burgdorferi pelos desafios utilizados isoladamente no presente estudo. O imunógeno rBm86 não causou oscilações significantes na produção de anticorpos IgG anti B. burgdorferi capazes de interferir nos resultados sorológicos para Borreliose de Lyme em bovinos. O estudo demonstrou a possibilidade de alterações transitórias na produção de anticorpos após estímulos vacinais e de estresse associados, ressaltando a necessidade de estudos sorológicos em conjunto a dados epidemiológicos e de manejo.

  4. Proving lipid rafts exist: membrane domains in the prokaryote Borrelia burgdorferi have the same properties as eukaryotic lipid rafts.

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    Timothy J LaRocca

    Full Text Available Lipid rafts in eukaryotic cells are sphingolipid and cholesterol-rich, ordered membrane regions that have been postulated to play roles in many membrane functions, including infection. We previously demonstrated the existence of cholesterol-lipid-rich domains in membranes of the prokaryote, B. burgdorferi, the causative agent of Lyme disease [LaRocca et al. (2010 Cell Host & Microbe 8, 331-342]. Here, we show that these prokaryote membrane domains have the hallmarks of eukaryotic lipid rafts, despite lacking sphingolipids. Substitution experiments replacing cholesterol lipids with a set of sterols, ranging from strongly raft-promoting to raft-inhibiting when mixed with eukaryotic sphingolipids, showed that sterols that can support ordered domain formation are both necessary and sufficient for formation of B. burgdorferi membrane domains that can be detected by transmission electron microscopy or in living organisms by Förster resonance energy transfer (FRET. Raft-supporting sterols were also necessary and sufficient for formation of high amounts of detergent resistant membranes from B. burgdorferi. Furthermore, having saturated acyl chains was required for a biotinylated lipid to associate with the cholesterol-lipid-rich domains in B. burgdorferi, another characteristic identical to that of eukaryotic lipid rafts. Sterols supporting ordered domain formation were also necessary and sufficient to maintain B. burgdorferi membrane integrity, and thus critical to the life of the organism. These findings provide compelling evidence for the existence of lipid rafts and show that the same principles of lipid raft formation apply to prokaryotes and eukaryotes despite marked differences in their lipid compositions.

  5. Ribotyping of Borrelia burgdorferi sensu lato in China%中国莱姆病螺旋体的核糖体基因分型研究

    Institute of Scientific and Technical Information of China (English)

    史翠霞; 万康林; 马凤琴; 张哲夫

    2001-01-01

    目的莱姆病螺旋体的基因型和临床表现、疫苗菌株和抗原菌株的选择存在密切的关系,所以对中国菌株进行分子流行病学研究,可为莱姆病的防治提供科学依据。方法 5S~23S rRNA基因间隔区RFLP分析和16+23S rRNA基因RFLP分析。结果中国菌株至少被分为3个基因型(B.burgdorferi sensu stricto、B.garinii和B.afzelii),B.garinii和B.afzelii占优势,B.burgdorferi sensu stricto少见。少数菌株用上述方法尚不能明确其分类地位,需进一步研究,中国很可能存在世界上独特的新基因型。结论中国菌株的基因型明显不同于北美菌株,而同欧洲菌株比较接近。5S~23S rRNA基因间隔区RFLP分析方法简便、快速、准确,是理想的基因型分类方法,可作为国内菌株基因型鉴定的常规方法。%Objective There is a close relationship between genospecies and clinical manifestation, the choice of strain for vaccine and serodiagnosis antigen. So molecular epidemiology of Borrelia burgdorferi sensu lato in China is important for the prevention of Lyme disease. Methods Rrf-rrl intergenic spacer RFLP analysis and 16+23S rRNA gene RFLP analysis. Results There were at least three genospecies in China. The most of them belongs to B. garinii and B. afzelii. A few isolates showed unique RFLP pattern. Their taxonomic positions remain unclear. There are probably new genospecies in China. Conclusions Chinese strains are quite different from that of Northern America and similar to that in Europe. The method of rrf-rrl intergenic spacer RFLP analysis is simple and accurate, so it could be used as routine genotyping method in China.

  6. Regulation of OspE-related, OspF-related, and Elp lipoproteins of Borrelia burgdorferi strain 297 by mammalian host-specific signals.

    Science.gov (United States)

    Hefty, P S; Jolliff, S E; Caimano, M J; Wikel, S K; Radolf, J D; Akins, D R

    2001-06-01

    In previous studies we have characterized the cp32/18 loci in Borrelia burgdorferi 297 which encode OspE and OspF orthologs and a third group of lipoproteins which possess OspE/F-like leader peptides (Elps). To further these studies, we have comprehensively analyzed their patterns of expression throughout the borrelial enzootic cycle. Serial dilution reverse transcription-PCR analysis indicated that although a shift in temperature from 23 to 37 degrees C induced transcription for all nine genes analyzed, this effect was often markedly enhanced in mammalian host-adapted organisms cultivated within dialysis membrane chambers (DMCs) implanted within the peritoneal cavities of rats. Indirect immunofluorescence assays performed on temperature-shifted, in vitro-cultivated spirochetes and organisms in the midguts of unfed and fed ticks revealed distinct expression profiles for many of the OspE-related, OspF-related, and Elp proteins. Other than BbK2.10 and ElpA1, all were expressed by temperature-shifted organisms, while only OspE, ElpB1, OspF, and BbK2.11 were expressed in the midguts of fed ticks. Additionally, although mRNA was detected for all nine lipoprotein-encoding genes, two of these proteins (BbK2.10 and ElpA1) were not expressed by spirochetes cultivated in vitro, within DMCs, or by spirochetes within tick midguts. However, the observation that B. burgdorferi-infected mice generated specific antibodies against BbK2.10 and ElpA1 indicated that these antigens are expressed only in the mammalian host and that a form of posttranscriptional regulation is involved. Analysis of the upstream regions of these genes revealed several differences between their promoter regions, the majority of which were found in the -10 and -35 hexamers and the spacer regions between them. Also, rather than undergoing simultaneous upregulation during tick feeding, these genes and the corresponding lipoproteins appear to be subject to progressive recruitment or enhancement of expression as

  7. HtrA, a Temperature- and Stationary Phase-Activated Protease Involved in Maturation of a Key Microbial Virulence Determinant, Facilitates Borrelia burgdorferi Infection in Mammalian Hosts.

    Science.gov (United States)

    Ye, Meiping; Sharma, Kavita; Thakur, Meghna; Smith, Alexis A; Buyuktanir, Ozlem; Xiang, Xuwu; Yang, Xiuli; Promnares, Kamoltip; Lou, Yongliang; Yang, X Frank; Pal, Utpal

    2016-08-01

    High-temperature requirement protease A (HtrA) represents a family of serine proteases that play important roles in microbial biology. Unlike the genomes of most organisms, that of Borrelia burgdorferi notably encodes a single HtrA gene product, termed BbHtrA. Previous studies identified a few substrates of BbHtrA; however, their physiological relevance could not be ascertained, as targeted deletion of the gene has not been successful. Here we show that BbhtrA transcripts are induced during spirochete growth either in the stationary phase or at elevated temperature. Successful generation of a BbhtrA deletion mutant and restoration by genetic complementation suggest a nonessential role for this protease in microbial viability; however, its remarkable growth, morphological, and structural defects during cultivation at 37°C confirm a high-temperature requirement for protease activation and function. The BbhtrA-deficient spirochetes were unable to establish infection of mice, as evidenced by assessment of culture, PCR, and serology. We show that transcript abundance as well as proteolytic processing of a borrelial protein required for cell fission and infectivity, BB0323, is impaired in BbhtrA mutants grown at 37°C, which likely contributed to their inability to survive in a mammalian host. Together, these results demonstrate the physiological relevance of a unique temperature-regulated borrelial protease, BbHtrA, which further enlightens our knowledge of intriguing aspects of spirochete biology and infectivity. PMID:27271745

  8. Epitope-Specific Evolution of Human B Cell Responses to Borrelia burgdorferi VlsE Protein from Early to Late Stages of Lyme Disease.

    Science.gov (United States)

    Jacek, Elzbieta; Tang, Kevin S; Komorowski, Lars; Ajamian, Mary; Probst, Christian; Stevenson, Brian; Wormser, Gary P; Marques, Adriana R; Alaedini, Armin

    2016-02-01

    Most immunogenic proteins of Borrelia burgdorferi, the causative agent of Lyme disease, are known or expected to contain multiple B cell epitopes. However, the kinetics of the development of human B cell responses toward the various epitopes of individual proteins during the course of Lyme disease has not been examined. Using the highly immunogenic VlsE as a model Ag, we investigated the evolution of humoral immune responses toward its immunodominant sequences in 90 patients with a range of early to late manifestations of Lyme disease. The results demonstrate the existence of asynchronous, independently developing, Ab responses against the two major immunogenic regions of the VlsE molecule in the human host. Despite their strong immunogenicity, the target epitopes were inaccessible to Abs on intact spirochetes, suggesting a lack of direct immunoprotective effect. These observations document the association of immune reactivity toward specific VlsE sequences with different phases of Lyme disease, demonstrating the potential use of detailed epitope mapping of Ags for staging of the infection, and offer insights regarding the pathogen's possible immune evasion mechanisms. PMID:26718339

  9. Comparative analysis of the infectivity rate of both Borrelia burgdorferi and Anaplasma phagocytophilum in humans and dogs in a New Jersey community

    Directory of Open Access Journals (Sweden)

    Gaito A

    2014-08-01

    Full Text Available Andrea Gaito,1 Vedrana Gjivoje,2 Sebastian Lutz,1 Ben Baxter2 1Private medical practice, Somerset County, NJ, USA; 2Bernardsville Animal Hospital, Somerset County, NJ, USA Abstract: Ticks are important vectors of disease and transmit an extensive array of bacterial, viral and protozoan diseases to both humans and dogs within a community. Borrelia burgdorferi, the causative agent of Lyme disease, has been extensively studied within both the human and veterinary population. Anaplasma phagocytophilum, an intracellular rickettsial pathogen also transmitted by ixodid ticks, has emerged as an important zoonotic infection with significant veterinary and medical implications, and is responsible for both canine granulocytic anaplasmosis and human granulocytic anaplasmosis. Multiple surveys exist in the international literature referencing infectivity rates of both of these diseases separately in both the dog and human populations. This is the first study to simultaneously examine the infectivity rate of both anaplasmosis and Lyme disease in humans and dogs in a community endemic for tick-borne diseases. Keywords: Lyme disease, anaplasmosis, dogs, humans 

  10. Acetyl-Phosphate Is Not a Global Regulatory Bridge between Virulence and Central Metabolism in Borrelia burgdorferi.

    Science.gov (United States)

    Richards, Crystal L; Lawrence, Kevin A; Su, Hua; Yang, Youyun; Yang, X Frank; Dulebohn, Daniel P; Gherardini, Frank C

    2015-01-01

    In B. burgdorferi, the Rrp2-RpoN-RpoS signaling cascade is a distinctive system that coordinates the expression of virulence factors required for successful transition between its arthropod vector and mammalian hosts. Rrp2 (BB0763), an RpoN specific response regulator, is essential to activate this regulatory pathway. Previous investigations have attempted to identify the phosphate donor of Rrp2, including the cognate histidine kinase, Hk2 (BB0764), non-cognate histidine kinases such as Hk1, CheA1, and CheA2, and small molecular weight P-donors such as carbamoyl-phosphate and acetyl-phosphate (AcP). In a report by Xu et al., exogenous sodium acetate led to increased expression of RpoS and OspC and it was hypothesized this effect was due to increased levels of AcP via the enzyme AckA (BB0622). Genome analyses identified only one pathway that could generate AcP in B. burgdorferi: the acetate/mevalonate pathway that synthesizes the lipid, undecaprenyl phosphate (C55-P, lipid I), which is essential for cell wall biogenesis. To assess the role of AcP in Rrp2-dependent regulation of RpoS and OspC, we used a unique selection strategy to generate mutants that lacked ackA (bb0622: acetate to AcP) or pta (bb0589: AcP to acetyl-CoA). These mutants have an absolute requirement for mevalonate and demonstrate that ackA and pta are required for cell viability. When the ΔackA or Δpta mutant was exposed to conditions (i.e., increased temperature or cell density) that up-regulate the expression of RpoS and OspC, normal induction of those proteins was observed. In addition, adding 20mM acetate or 20mM benzoate to the growth media of B. burgdorferi strain B31 ΔackA induced the expression of RpoS and OspC. These data suggest that AcP (generated by AckA) is not directly involved in modulating the Rrp2-RpoN-RpoS regulatory pathway and that exogenous acetate or benzoate are triggering an acid stress response in B. burgdorferi. PMID:26681317

  11. Acetyl-Phosphate Is Not a Global Regulatory Bridge between Virulence and Central Metabolism in Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Crystal L Richards

    Full Text Available In B. burgdorferi, the Rrp2-RpoN-RpoS signaling cascade is a distinctive system that coordinates the expression of virulence factors required for successful transition between its arthropod vector and mammalian hosts. Rrp2 (BB0763, an RpoN specific response regulator, is essential to activate this regulatory pathway. Previous investigations have attempted to identify the phosphate donor of Rrp2, including the cognate histidine kinase, Hk2 (BB0764, non-cognate histidine kinases such as Hk1, CheA1, and CheA2, and small molecular weight P-donors such as carbamoyl-phosphate and acetyl-phosphate (AcP. In a report by Xu et al., exogenous sodium acetate led to increased expression of RpoS and OspC and it was hypothesized this effect was due to increased levels of AcP via the enzyme AckA (BB0622. Genome analyses identified only one pathway that could generate AcP in B. burgdorferi: the acetate/mevalonate pathway that synthesizes the lipid, undecaprenyl phosphate (C55-P, lipid I, which is essential for cell wall biogenesis. To assess the role of AcP in Rrp2-dependent regulation of RpoS and OspC, we used a unique selection strategy to generate mutants that lacked ackA (bb0622: acetate to AcP or pta (bb0589: AcP to acetyl-CoA. These mutants have an absolute requirement for mevalonate and demonstrate that ackA and pta are required for cell viability. When the ΔackA or Δpta mutant was exposed to conditions (i.e., increased temperature or cell density that up-regulate the expression of RpoS and OspC, normal induction of those proteins was observed. In addition, adding 20mM acetate or 20mM benzoate to the growth media of B. burgdorferi strain B31 ΔackA induced the expression of RpoS and OspC. These data suggest that AcP (generated by AckA is not directly involved in modulating the Rrp2-RpoN-RpoS regulatory pathway and that exogenous acetate or benzoate are triggering an acid stress response in B. burgdorferi.

  12. Use of the "blue halo" assay in the identification of genes encoding exported proteins with cleavable signal peptides: cloning of a Borrelia burgdorferi plasmid gene with a signal peptide.

    Science.gov (United States)

    Giladi, M; Champion, C I; Haake, D A; Blanco, D R; Miller, J F; Miller, J N; Lovett, M A

    1993-07-01

    We have recently reported a phoA expression vector, termed pMG, which, like TnphoA, is useful in identifying genes encoding membrane-spanning sequences or signal peptides. This cloning system has been modified to facilitate the distinction of outer membrane and periplasmic alkaline phosphatase (AP) fusion proteins from inner membrane AP fusion proteins by transforming pMG recombinants into Escherichia coli KS330, the strain utilized in the "blue halo" assay first described by Strauch and Beckwith (Proc. Natl. Acad. Sci. USA 85:1576-1580, 1988). The lipoprotein mutation lpp-5508 of KS330 results in an outer membrane that is leaky to macromolecules, and its degP4 mutation greatly reduces periplasmic proteolytic degradation of AP fusion proteins. pMG AP fusions containing cleavable signal peptides, including the E. coli periplasmic protein beta-lactamase, the E. coli and Chlamydia trachomatis outer membrane proteins OmpA and MOMP, respectively, and Tp 9, a Treponema pallidum AP recombinant, diffused through the leaky outer membrane of KS330 and resulted in blue colonies with blue halos. In contrast, inner membrane AP fusions derived from E. coli proteins, including leader peptidase, SecY, and the tetracycline resistance gene product, as well as Tp 70, a T. pallidum AP recombinant which does not contain a signal peptide, resulted in blue colonies without blue halos. Lipoprotein-AP fusions, including the Borrelia burgdorferi OspA and T. pallidum Tp 75 and TmpA showed halo formation, although there was significantly less halo formation than that produced by either periplasmic or outer membrane AP fusions. In addition, we applied this approach to screen recombinants constructed from a 9.0-kb plasmid isolated from the B31 virulent strain of B. burgdorferi. One of the blue halo colonies identified produced an AP fusion protein which contained a signal peptide with a leader peptidase I cleavage recognition site. The pMG/KS330r- cloning and screening approach can identify

  13. Mapping human risk of infection with Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, in a periurban forest in France.

    Science.gov (United States)

    Vourc'h, G; Abrial, D; Bord, S; Jacquot, M; Masséglia, S; Poux, V; Pisanu, B; Bailly, X; Chapuis, J-L

    2016-07-01

    Lyme borreliosis is a major zoonosis in Europe, with estimates of over 26,000 cases per year in France alone. The etiological agents are spirochete bacteria that belong to the Borrelia burgdorferi sensu lato (s. l.) complex and are transmitted by hard ticks among a large range of vertebrate hosts. In Europe, the tick Ixodes ricinus is the main vector. In the absence of a vaccine and given the current difficulties to diagnose and treat chronic Lyme syndromes, there is urgent need for prevention. In this context, accurate information on the spatial patterns of risk of exposure to ticks is of prime importance for public health. The objective of our study was to provide a snapshot map of the risk of human infection with B. burgdorferi s. l. pathogens in a periurban forest at a high resolution, and to analyze the factors that contribute to variation in this risk. Field monitoring took place over three weeks in May 2011 in the suburban Sénart forest (3,200ha; southeast of Paris), which receives over 3 million people annually. We sampled ticks over the entire forest area (from 220 forest stands with a total area of 35,200m(2)) and quantified the density of questing nymphs (DON), the prevalence of infection among nymphs (NIP), and the density of infected nymphs (DIN), which is the most important predictor of the human risk of Lyme borreliosis. For each of these response variables, we explored the relative roles of weather (saturation deficit), hosts (abundance indices of ungulates and Tamias sibiricus, an introduced rodent species), vegetation and forest cover, superficial soil composition, and the distance to forest roads. In total, 19,546 questing nymphs were collected and the presence of B. burgdorferi s. l. was tested in 3,903 nymphs by qPCR. The mean DON was 5.6 nymphs per 10m(2) (standard deviation=10.4) with an average NIP of 10.1% (standard deviation=0.11). The highest DIN was 8.9 infected nymphs per 10m(2), with a mean of 0.59 (standard deviation=0.6). Our

  14. 莱姆病螺旋体鞭毛平截性蛋白的表达及其诊断潜力研究%Expression on specific fragement of flagellin from Borrelia burgdorferi and its diagnostic

    Institute of Scientific and Technical Information of China (English)

    仝彩玲; 吴银娟; 周勇志; 曹杰; 李培英; 周金林

    2011-01-01

    目的 原核表达伯氏疏螺旋体鞭毛蛋白Flagellin A基因特异性区段,获得重组鞭毛蛋白平截性蛋白作为诊断抗原,建立间接ELISA方法用于动物莱姆病的诊断.方法 PCR扩增获取伯氏疏螺旋体鞭毛蛋白基因的同源性较低的第394-798 bp区段,构建重组质粒pGEX-4T-(1)/tFlaA,构建好的表达质粒转化到大肠杆菌BL21( DE3)中进行表达,并纯化重组蛋白,用纯化的表达蛋白作为莱姆病诊断的抗原,用于ELISA检测实验感染小鼠莱姆病.结果 成功构建莱姆病螺旋体鞭毛平截性蛋白的表达载体,重组蛋白在宿主菌内高效、稳定表达,重组平截性蛋白显示了可作为ELISA诊断的抗原用于莱姆病的诊断价值.结论 纯化的伯氏疏螺旋体鞭毛蛋白可作为莱姆病ELISA诊断抗原用于莱姆病的诊断,为莱姆病快速诊断试剂盒的开发打下基础.%The purpose was to extract Borrelia burgdorferi DNA and use PCR method to obtain the gene coding of the Borrelia burgdorferi flagellin from 394 to 798 nucleotide that exhibited low homology with related gene from other bactdrial specises we insert Borrelia burgdorferi flagellin from 394 to 798 nucleotide to PGEX-4T-1 vector, verified by DNA sequence detection and tren thansformed it into in E. coli BL 21 (DE3) to induce target protein. Then we purified the recombination pro-tein as ELISA antigen to diagnosis Lyme disease. On other hand, we infected mouse with Borrelia burgdorferi. Two weeks, we got positive serum from mouse. We used recombinant protein as diagnostic antigen andpositive mouse serum as antibody to con-duct ELISA test and analysts. The ELISA results showed the feasibility of recombinant protein as diagnostic antigen to diag-nose Lyme disease, which lay foundations for the diagnosis of Lyme disease.

  15. Enhancement of immune response towards non-lipidized Borrelia burgdorferi recombinant OspC antigen by binding onto the surface of metallochelating nanoliposomes with entrapped lipophilic derivatives of norAbuMDP

    Czech Academy of Sciences Publication Activity Database

    Křupka, M.; Mašek, J.; Bartheldyová, E.; Turánek Knotigová, P.; Plocková, J.; Korvasová, Z.; Škrabalová, M.; Koudelka, Š.; Kulich, P.; Zachová, K.; Czerneková, L.; Strouhal, O.; Horynová, M.; Šebela, M.; Miller, A. D.; Ledvina, Miroslav; Raška, M.; Turánek, J.

    2012-01-01

    Roč. 160, č. 2 (2012), s. 374-381. ISSN 0168-3659 R&D Projects: GA ČR(CZ) GAP304/10/1951; GA TA ČR TA01011165; GA AV ČR(CZ) KAN200520703; GA AV ČR KAN200100801 Grant ostatní: GA UP(CZ) LF-2010-014; GA UP(CZ) LF-2011-002 Institutional research plan: CEZ:AV0Z40550506 Keywords : Borrelia burgdorferi * OspC * metallochelating liposomes * MDP Subject RIV: CC - Organic Chemistry Impact factor: 7.633, year: 2012

  16. Functional study of Treponema pallidum genes using Borrelia burgdorferi as a surrogate system%以伯氏疏螺旋体为载体菌研究梅毒螺旋体的基因功能

    Institute of Scientific and Technical Information of China (English)

    尹燕萍; 史玢; 唐志德; 楼永良; 杨晓峰

    2015-01-01

    目的:利用体外可培养的伯氏疏螺旋体作为载体菌,将梅毒螺旋体的基因转入相应的伯氏疏螺旋体菌株进行功能性表达,进而对梅毒螺旋体的基因功能进行研究。方法构建一个携带梅毒螺旋体基因tp0111的大肠埃希菌-伯氏疏螺旋体穿梭质粒,利用电转化法,将质粒转入相应的伯氏疏螺旋体菌株中,通过观察伯氏疏螺旋体的表型和相关基因表达变化,来研究梅毒螺旋体的基因功能。结果成功将梅毒螺旋体基因tp0111转入伯氏疏螺旋体rpoN突变株内,一定程度上回补了伯氏疏螺旋体突变株的表型缺陷。结论首次证明tp0111是梅毒螺旋体的rpoN基因,同时验证了伯氏疏螺旋体可以作为载体来研究梅毒螺旋体的致病因子及其基因调控系统。%Objective To employ Borrelia burgdorferi( B. burgdorferi) , a culturable and genetical-ly transformable spirochete, as a surrogate system to study Treponema pallidum ( T. pallidum) gene function. Methods Bioinformatic analysis revealed that the T. pallidum gene tp0111 encodes the putative sigma factor RpoN. We constructed a B. burgdorferi shuttle vector harboring tp0111. The shuttle vector was then trans-formed into the B. burgdorferi rpoN mutant strain. The phenotype of the resulting B. burgdorferi strain was then determined. Results We successfully constructed the B. burgdorferi rpoN mutant carrying the T. palli-dum gene tp0111. We found that tp0111 could partially complement the B. burgdorferi rpoN mutant. Con-clusion This work provides the first experimental evidence showing that tp0111 is the rpoN gene of T. palli-dum. It also demonstrates that B. burgdorferi can be used as a surrogate system for studying T. pallidum gene function.

  17. Phagosomal signaling by Borrelia burgdorferi in human monocytes involves Toll-like receptor (TLR) 2 and TLR8 cooperativity and TLR8-mediated induction of IFN-β

    Science.gov (United States)

    Cervantes, Jorge L.; Dunham-Ems, Star M.; La Vake, Carson J.; Petzke, Mary M.; Sahay, Bikash; Sellati, Timothy J.; Radolf, Justin D.; Salazar, Juan C.

    2011-01-01

    Phagocytosed Borrelia burgdorferi (Bb) induces inflammatory signals that differ both quantitatively and qualitatively from those generated by spirochetal lipoproteins interacting with Toll-like receptor (TLR) 1/2 on the surface of human monocytes. Of particular significance, and in contrast to lipoproteins, internalized spirochetes induce transcription of IFN-β. Using inhibitory immunoregulatory DNA sequences (IRSs) specific to TLR7, TLR8, and TLR9, we show that the TLR8 inhibitor IRS957 significantly diminishes production of TNF-α, IL-6, and IL-10 and completely abrogates transcription of IFN-β in Bb-stimulated monocytes. We demonstrate that live Bb induces transcription of TLR2 and TLR8, whereas IRS957 interferes with their transcriptional regulation. Using confocal and epifluorescence microscopy, we show that baseline TLR expression in unstimulated monocytes is greater for TLR2 than for TLR8, whereas expression of both TLRs increases significantly upon stimulation with live spirochetes. By confocal microscopy, we show that TLR2 colocalization with Bb coincides with binding, uptake, and formation of the phagosomal vacuole, whereas recruitment of both TLR2 and TLR8 overlaps with degradation of the spirochete. We provide evidence that IFN regulatory factor (IRF) 7 is translocated into the nucleus of Bb-infected monocytes, suggesting its activation through phosphorylation. Taken together, these findings indicate that the phagosome is an efficient platform for the recognition of diverse ligands; in the case of Bb, phagosomal signaling involves a cooperative interaction between TLR2 and TLR8 in pro- and antiinflammatory cytokine responses, whereas TLR8 is solely responsible for IRF7-mediated induction of IFN-β. PMID:21321205

  18. Activation of human monocytes by live Borrelia burgdorferi generates TLR2-dependent and -independent responses which include induction of IFN-beta.

    Directory of Open Access Journals (Sweden)

    Juan C Salazar

    2009-05-01

    Full Text Available It is widely believed that innate immune responses to Borrelia burgdorferi (Bb are primarily triggered by the spirochete's outer membrane lipoproteins signaling through cell surface TLR1/2. We recently challenged this notion by demonstrating that phagocytosis of live Bb by peripheral blood mononuclear cells (PBMCs elicited greater production of proinflammatory cytokines than did equivalent bacterial lysates. Using whole genome microarrays, we show herein that, compared to lysates, live spirochetes elicited a more intense and much broader transcriptional response involving genes associated with diverse cellular processes; among these were IFN-beta and a number of interferon-stimulated genes (ISGs, which are not known to result from TLR2 signaling. Using isolated monocytes, we demonstrated that cell activation signals elicited by live Bb result from cell surface interactions and uptake and degradation of organisms within phagosomes. As with PBCMs, live Bb induced markedly greater transcription and secretion of TNF-alpha, IL-6, IL-10 and IL-1beta in monocytes than did lysates. Secreted IL-18, which, like IL-1beta, also requires cleavage by activated caspase-1, was generated only in response to live Bb. Pro-inflammatory cytokine production by TLR2-deficient murine macrophages was only moderately diminished in response to live Bb but was drastically impaired against lysates; TLR2 deficiency had no significant effect on uptake and degradation of spirochetes. As with PBMCs, live Bb was a much more potent inducer of IFN-beta and ISGs in isolated monocytes than were lysates or a synthetic TLR2 agonist. Collectively, our results indicate that the enhanced innate immune responses of monocytes following phagocytosis of live Bb have both TLR2-dependent and -independent components and that the latter induce transcription of type I IFNs and ISGs.

  19. Abrogation of ospAB constitutively activates the Rrp2-RpoN-RpoS pathway (sigmaN-sigmaS cascade) in Borrelia burgdorferi.

    Science.gov (United States)

    He, Ming; Oman, Tara; Xu, Haijun; Blevins, Jon; Norgard, Michael V; Yang, X Frank

    2008-12-01

    Molecular mechanisms underlying the reciprocal regulation of the two major surface lipoproteins and virulence factors of Borrelia burgdorferi, OspA and OspC, are not fully understood. Herein, we report that inactivation of the ospAB operon resulted in overproduction of OspC and many other lipoproteins via the constitutive activation of the Rrp2-RpoN-RpoS pathway. Complementing the ospAB mutant with a wild-type copy of ospA, but not an ospA variant that lacks the lipoprotein signal sequence, restored normal regulation of the Rrp2-RpoN-RpoS pathway; these results indicate that the phenotype was not caused by spurious mutations. Interestingly, while most of the ospAB mutant clones displayed a constitutive ospC expression phenotype, some ospAB mutant clones showed little or no ospC expression. Further analyses revealed that this OspC-negative phenotype was independent of abrogation of ospAB. While activation of the Rrp2-RpoN-RpoS pathway was recently shown to downregulate ospA, our findings suggest that reduction of OspA can also activate this pathway. We postulate that the activation of the Rrp2-RpoN-RpoS pathway and downregulation of OspA form a positive feedback loop that allows spirochaetes to produce and maintain a constant high level of OspC and other lipoproteins during tick feeding, a strategy that is critical for spirochaetal transmission and mammalian infection. PMID:19019147

  20. 莱姆病螺旋体重组质粒pBX1的DNA序列分析及其在大肠杆菌中的诱导表达%DNA Sequence Analysis and Expression of the Recombinant Plasmid pBX1 from Borrelia Burgdorferi B31 Strain

    Institute of Scientific and Technical Information of China (English)

    谢勇恩; 鲍朗; 胡昌华; 李学敏; 陈炜

    2001-01-01

    after induced with IPTG.The recombinant fusion protein could be recongnized by rabbit polyclonal antiserum against Borrelia burgdorferi B31 strain. Conclusion A recombinant plasmid which contains the gene fragment encoding the specific region of the 83kd antigen protein from Borrelia burgdorferi B31 strain has been successfully constructed.The recombinant plasmid can stably express 29kd fusion protein in E.coli XL1-Blue MRF′.These results could serve as a base of further studies on the usefulness of the fusion protein in serodiagnosis and vaccine for Lyme disease.

  1. Isolation and identification of Borrelia burgdorferi sensu lato from ticks in six provinces in China%中国六省蜱中莱姆病螺旋体分离与鉴定

    Institute of Scientific and Technical Information of China (English)

    耿震; 侯学霞; 万康林; 郝琴

    2010-01-01

    目的 了解中国6省蜱中莱姆病螺旋体携带情况.方法 在6省各选取2个采样点捕蜱,采用病原分离培养和巢式PCR方法对蜱进行莱姆病螺旋体检测,通过基因测序确定分型.结果 6省共采集到2200余只蜱,约1000只蜱用于病原分离培养,从吉林省长白县全沟硬蜱标本中分离到13株螺旋体,从贵州省道真粒形硬蜱标本中分离到9株螺旋体.1255只蜱用于PCR检测,从6省的蜱标本中检测到莱姆病螺旋体特异片段,其中吉林省(长白县27.08%、通化县20.41%)、青海省(互助县25.06%、黄南县21.11%)和贵州省(道真县25.63%)蜱中莱姆病螺旋体阳性率较高,山西省(垣曲县4.72%、交城县3.64%)蜱中莱姆病螺旋体阳性率较低.通过序列同源性分析确定吉林、青海、甘肃和山西省蜱中莱姆病螺旋体基因型均为Borrelia garinii.贵州、湖南省的基因型均为Borrelia valaisiana.结论 6省蜱中均带有莱姆病螺旋体,且带菌率有差异;山西省蜱中存在Borrelia ganmi型莱姆病螺旋体,湖南省蜱中存在Borrelia valaisiana型莱姆病螺旋体.%Objective To understand the carrying status of Borrelia burgdorferi in ticks from the mountain areas from six representative provinces, including Jilin, Shanxi, Gansu, Qinghai,Guizhou and Hunan in China. Methods Flagging and trapping methods were used to collect ticks in forest area and culture medium was used to isolate the pathogen. Nested-PCR was used to detect the gem-carrying rate of ticks. Results More than 2200 ticks from six representative provinces were collected and 1000 ticks were used to isolate the pathogen. 13 Lyme disease spirochetes from ixodes persulcatus in Changbai, Jilin province and 9 Lyme disease spirochetes from ixodes granulatus in Daozhen, Guizhou province were identified. There were 1255 ticks used for PCR testing. Specific fragments of the Borrelia burgdorferi in ticks were found from the six representative provinces in China. The carrier

  2. Larvae of Ixodes ricinus transmit Borrelia afzelii and B. miyamotoi to vertebrate hosts

    NARCIS (Netherlands)

    Duijvendijk, van Gilian; Coipan, Claudia; Wagemakers, Alex; Fonville, Manoj; Ersöz, Jasmin; Oei, Anneke; Földvári, Gábor; Hovius, Joppe; Takken, Willem; Sprong, Hein

    2016-01-01

    Background: Lyme borreliosis is the most common tick-borne human disease and is caused by Borrelia burgdorferi sensu lato (s.l.). Borrelia miyamotoi, a relapsing fever spirochaete, is transmitted transovarially, whereas this has not been shown for B. burgdorferi (s.l). Therefore, B. burgdorferi (

  3. Mutations in the Borrelia burgdorferi Flagellar Type III Secretion System Genes fliH and fliI Profoundly Affect Spirochete Flagellar Assembly, Morphology, Motility, Structure, and Cell Division

    Science.gov (United States)

    Gao, Lihui; Zhao, Xiaowei; Liu, Jun; Norris, Steven J.

    2015-01-01

    ABSTRACT The Lyme disease spirochete Borrelia burgdorferi migrates to distant sites in the tick vectors and mammalian hosts through robust motility and chemotaxis activities. FliH and FliI are two cytoplasmic proteins that play important roles in the type III secretion system (T3SS)-mediated export and assembly of flagellar structural proteins. However, detailed analyses of the roles of FliH and FliI in B. burgdorferi have not been reported. In this study, fliH and fliI transposon mutants were utilized to dissect the mechanism of the Borrelia type III secretion system. The fliH and fliI mutants exhibited rod-shaped or string-like morphology, greatly reduced motility, division defects (resulting in elongated organisms with incomplete division points), and noninfectivity in mice by needle inoculation. Mutants in fliH and fliI were incapable of translational motion in 1% methylcellulose or soft agar. Inactivation of either fliH or fliI resulted in the loss of the FliH-FliI complex from otherwise intact flagellar motors, as determined by cryo-electron tomography (cryo-ET). Flagellar assemblies were still present in the mutant cells, albeit in lower numbers than in wild-type cells and with truncated flagella. Genetic complementation of fliH and fliI mutants in trans restored their wild-type morphology, motility, and flagellar motor structure; however, full-length flagella and infectivity were not recovered in these complemented mutants. Based on these results, disruption of either fliH or fliI in B. burgdorferi results in a severe defect in flagellar structure and function and cell division but does not completely block the export and assembly of flagellar hook and filament proteins. PMID:25968649

  4. Isolation of Borrelia spirochetes from patients in Texas.

    OpenAIRE

    Rawlings, J A; Fournier, P V; Teltow, G. J.

    1987-01-01

    The Texas Department of Health Laboratory began culturing the Lyme disease spirochete Borrelia burgdorferi in 1985. This organism was subsequently isolated from blood, cerebrospinal fluid, joint fluid, skin, bone, and autopsy tissues from humans. Fluorescent-antibody tests with murine monoclonal antibodies confirmed that seven of these isolates were B. burgdorferi and that two others belonged to the genus Borrelia.

  5. 一种伯氏疏螺旋体表达质粒的构建%Construction of a shuttle vector for inducible gene expression in Borrelia burgdorferi

    Institute of Scientific and Technical Information of China (English)

    叶美萍; 黄龙丽; 庄振超; 楼永良

    2014-01-01

    Objective To construct a shuttle plasmid for inducible gene expression in Borrelia burgdorferi (B.burgdorferi) with an advantage of flexible genetic manipulation.Methods The IPTG-inducible lac repressor/operator system from Escherichia coli (E.coli) was adopted and modified in the current study.The plasmid shuttle vector was developed by inserting multiple cloning sites,FLAG and HA tags into the shuttle vector by molecular cloning approaches.The target gene was inserted at the site under the control of the promoter (Tn5 derivate) in plasmid pQE30.This promoter contained two lac operators and a codonoptimized lacI gene driven by flaB promoter.Results A plasmid shuttle vector,pJJ275,was successfully constructed with the ability to express target genes in B.burgdorferi in the presence of IPTG.By using this system,a HA-tagged rpoS gene was introduced into the typical infectious strain B.burgdorferi B31.The target gene expression induced by IPTG was confirmed at transcriptional and translational levels.The RpoS dependent virulence factor of Borrelia,OspC,was also detected,indicating that the expressed protein was functional.Conclusion The constructed plasmid shuttle vector can express exogenous genes in B.burgdorferi with an inducible feature and an advantage of flexible genetic manipulation.It can be applied for genetic manipulation of B.burgdorferi involved in gene regulation and complementation.%目的 构建一个具有较强遗传可操作性的大肠杆菌-伯氏疏螺旋体表达穿梭质粒,以期作为工具质粒在疏螺旋体中实现外源基因的可诱导表达.方法 利用源自大肠杆菌的lac表达/诱导系统,在已有的穿梭质粒的基础上,通过分子生物学技术加入多克隆位点和HA、FLAG蛋白表达标签,增加其遗传可操作性.结果 成功构建工具质粒pJ J275,可以用于伯氏疏螺旋体的基因可控制表达.以rpoS基因为例,将其克隆至pJJ275并转入疏螺旋体中,获得的菌株在IPTG存在的条件

  6. The crystal structures of two salivary cystatins from the tick Ixodes scapularis and the effect of these inhibitors on the establishment of Borrelia burgdorferi infection in a murine model

    Energy Technology Data Exchange (ETDEWEB)

    Kotsyfakis, Michalis; Horka, Helena; Salat, Jiri; Andersen, John F. (South Bohemia); (ASCR-ICP); (NIAID)

    2010-11-17

    We have previously demonstrated that two salivary cysteine protease inhibitors from the Borrelia burgdorferi (Lyme disease) vector Ixodes scapularis - namely sialostatins L and L2 - play an important role in tick biology, as demonstrated by the fact that silencing of both sialostatins in tandem results in severe feeding defects. Here we show that sialostatin L2 - but not sialostatin L - facilitates the growth of B. burgdorferi in murine skin. To examine the structural basis underlying these differential effects of the two sialostatins, we have determined the crystal structures of both sialostatin L and L2. This is the first structural analysis of cystatins from an invertebrate source. Sialostatin L2 crystallizes as a monomer with an 'unusual' conformation of the N-terminus, while sialostatin L crystallizes as a domain-swapped dimer with an N-terminal conformation similar to other cystatins. Deletion of the 'unusual' N-terminal five residues of sialostatin L2 results in marked changes in its selectivity, suggesting that this region is a particularly important determinant of the biochemical activity of sialostatin L2. Collectively, our results reveal the structure of two tick salivary components that facilitate vector blood feeding and that one of them also supports pathogen transmission to the vertebrate host.

  7. Serological survey of Bartonella spp., Borrelia burgdorferi, Brucella spp., Coxiella burnetii, Francisella tularensis, Leptospira spp., Echinococcus, Hanta-, TBE- and XMR-virus infection in employees of two forestry enterprises in North Rhine-Westphalia, Germany, 2011-2013.

    Science.gov (United States)

    Jurke, Annette; Bannert, N; Brehm, K; Fingerle, V; Kempf, V A J; Kömpf, D; Lunemann, M; Mayer-Scholl, A; Niedrig, M; Nöckler, K; Scholz, H; Splettstoesser, W; Tappe, D; Fischer, Silke F

    2015-10-01

    We initiated a survey to collect basic data on the frequency and regional distribution of various zoonoses in 722 employees of forestry enterprises in the German state of North Rhine-Westphalia (NRW) from 2011 to 2013. Exposures associated with seropositivity were identified to give insight into the possible risk factors for infection with each pathogen. 41.2% of participants were found to be seropositive for anti-Bartonella IgG, 30.6% for anti-Borrelia burgdorferi IgG, 14.2% for anti-Leptospira IgG, 6.5% for anti-Coxiella burnetii IgG, 6.0% for anti-Hantavirus IgG, 4.0% for anti-Francisella tularensis IgG, 3.4% for anti-TBE-virus IgG, 1.7% for anti-Echinococcus IgG, 0.0% for anti-Brucella IgG and anti-XMRV IgG. Participants seropositive for B. burgdorferi were 3.96 times more likely to be professional forestry workers (univariable analysis: OR 3.96; 95% CI 2.60-6.04; pforestry workers nor office workers represent a risk population and that NRW is not a typical endemic area. Forestry workers appear to have higher risk for contact with B. burgdorferi-infected ticks and a regionally diverse risk for acquiring Hantavirus-infection. The regional epidemiology of zoonoses is without question of great importance for public health. Knowledge of the regional risk factors facilitates the development of efficient prevention strategies and the implementation of such prevention measures in a sustainable manner. PMID:26422407

  8. Use of CFSE staining of borreliae in studies on the interaction between borreliae and human neutrophils

    Directory of Open Access Journals (Sweden)

    Hytönen Jukka

    2006-10-01

    Full Text Available Abstract Background Species of the tick-transmitted spirochete group Borrelia burgdorferi sensu lato (B. burgdorferi cause Lyme borreliosis. Acute borrelial infection of the skin has unusual characteristics with only a mild local inflammatory response suggesting that the interaction between borreliae and the cells of the first-line defence might differ from that of other bacteria. It has been reported that human neutrophils phagocytose motile borreliae through an unconventional mechanism (tube phagocytosis which is not observed with non-motile borreliae. Therefore, it would be of great interest to visualise the bacteria by a method not affecting motility and viability of borreliae to be able to study their interaction with the cells of the innate immunity. Carboxyfluorescein diacetate, succinimidyl ester (CFSE labelling has been previously used for studying the adhesion of labelled bacteria to host cells and the uptake of labelled substrates by various cells using flow cytometry. Results In this study, CFSE was shown to efficiently stain different genospecies of B. burgdorferi without affecting bacterial viability or motility. Use of CFSE staining allowed subsequent quantification of borreliae associated with human neutrophils with flow cytometry and confocal microscopy. As a result, no difference in association between different borrelial genospecies (Borrelia burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, or between borreliae and the pyogenic bacterium Streptococcus pyogenes, with neutrophils could be detected. Borrelial virulence, on the other hand, affected association with neutrophils, with significantly higher association of a non-virulent mutant B. burgdorferi sensu stricto strain compared to the parental virulent wild type strain. Conclusion These results suggest that the flow cytometric assay using CFSE labelled borreliae is a valuable tool in the analysis of the interaction between borreliae and human neutrophils. The

  9. The ospA genes of Borrelia burgdorferi sensu lato detected in rodents in Zhejiang province%浙江省在鼠类中检测到伯氏疏螺旋体ospA基因

    Institute of Scientific and Technical Information of China (English)

    姜理平; 陆群英; 李钟梁; 张磊; 王复甦; 张政

    2012-01-01

    目的 了解浙江省磐安县鼠中感染伯氏疏螺旋体的情况,研究以外膜蛋白A(OspA)为靶基因的PCR方法在浙江省的应用.方法 利用PCR方法,检测磐安县128份鼠肝、脾标本中ospA基因特异片段;对所检测到的阳性结果进行序列测定,并进行分析.结果 从128份鼠肝、脾标本中检测发现4例ospA基因阳性片段,其中3份来自黄毛鼠,1份来自大林姬鼠;核酸序列基本一致,与伯氏疏螺旋体法雷斯疏螺旋体高度相似.结论 利用ospA基因能从标本中检测到伯氏疏螺旋体;浙江省部分山区存在鼠感染伯氏疏螺旋体的情况,其中伯氏疏螺旋体法雷斯疏螺旋体占优势.%Objective To identify Borrelia burgdorferi sensu lato infection of rodents in Pan' an county, Zhejiang province. Methods The ospA gene specific fragments from 128 mouse liver and spleen samples were detected using the PCR method and the positive samples were sequenced, followed by phylogenetic analysis. Results It was found that there were four ospA gene positive fragments out of the 128 samples, of which 3 fragments were obtained from Rattus losea and the other one was from Apodemus peninsulae. The 4 positive fragments shared high similarity with each other in terms of the sequences and shared high identities with B. Valaisiana. Conclusion B. Burgdorferi sensu lato can be detected from samples using the ospA gene. There exists B. Burgdorferi sensu lato infection in rodents in some mountain areas of Zhejiang province, and B. Valaisiana is found to be predominant.

  10. Invasive potential of Borrelia burgdorferi sensu stricto ospC type L strains increases the possible disease risk to humans in the regions of their distribution

    Czech Academy of Sciences Publication Activity Database

    Golovchenko, Maryna; Šíma, Radek; Hajdušek, Ondřej; Grubhoffer, Libor; Oliver, J. H., Jr.; Rudenko, Natalia

    2014-01-01

    Roč. 7, NOV 28 2014 (2014), s. 538. ISSN 1756-3305 R&D Projects: GA ČR GP13-27630P; GA ČR GP13-12816P Institutional support: RVO:60077344 Keywords : B. burgdorferi ospC type * Invasive potential * Lyme disease * Southeastern U.S.A. * Tick vector * Vertebrate host Subject RIV: EC - Immunology Impact factor: 3.430, year: 2014

  11. Immunochemical characterization of and isolation of the gene for a Borrelia burgdorferi immunodominant 60-kilodalton antigen common to a wide range of bacteria

    DEFF Research Database (Denmark)

    Hansen, K; Bangsborg, Jette Marie; Fjordvang, H;

    1988-01-01

    . burgdorferi CA was localized on a 2.3-kilobase fragment of the plasmid pKH1. CA may have pathogenetic implications in Lyme borreliosis, since the CA of mycobacteria recently has been shown to play a role in the etiology of experimental autoimmune arthritis. The extensive cross-reactivity of this antigen may...... account for the low diagnostic specificity of the currently used serological tests in Lyme borreliosis....

  12. A serological survey of tick-borne pathogens in dogs in North America and the Caribbean as assessed by Anaplasma phagocytophilum, A. platys, Ehrlichia canis, E. chaffeensis, E. ewingii, and Borrelia burgdorferi species-specific peptides

    Directory of Open Access Journals (Sweden)

    Barbara A. Qurollo

    2014-10-01

    Full Text Available Introduction: Tick-borne pathogens cause a spectrum of disease manifestations in both dogs and humans. Recognizing regional and temporal shifts in exposure are important as tick distributions change. To better delineate regional exposure to canine tick-borne pathogens, an expanded set of species-specific peptides were used to detect Anaplasma phagocytophilum (Aph, Anaplasma platys (Apl, Ehrlichia canis (Ec, Ehrlichia chaffeensis (Ech, Ehrlichia ewingii (Eew, and Borrelia burgdorferi (Bb antibodies in canine serum. Methods: Archived canine serum samples (n=6,582 collected during 2008–2010 and in 2012 from the US, Canada, and the Caribbean were retrospectively screened for antibodies against Ehrlichia and Anaplasma species-specific peptides. Overall, regional and temporal seroprevalence rates were determined. Results: Overall Bb and Eew were the most seroprevalent pathogens. During 2008–2010, seroprevalence rates increased overall for Aph and Ech, and regionally, Bb and Aph seroprevalence rates increased in the South. Canada had unexpectedly high seroprevalence rates for Ec and Apl. The most common co-exposures were Eew+Ech, followed by Aph+Bb and Eew+Bb. Conclusions: This study demonstrated significant shifts in canine vector-borne disease seroprevalence rates. The use of specific peptides facilitated improved geographic delineation of tick-borne pathogen distributions among dogs, which may enhance epidemiological surveillance of vector-borne pathogens shared by dogs and humans.

  13. A Genotype Loop-mediated Isothermal Amplification (LAMP) for Borrelia Burgdorferi of Lyme Disease%环介导恒温扩增对莱姆病病原伯氏疏螺旋体的分型鉴定

    Institute of Scientific and Technical Information of China (English)

    杨吉飞; 杨朝霞; 罗建勋; 殷宏; 关贵全; 牛庆丽; 李有全; 刘军龙; 马米玲; 任巧云; 刘爱红; 王子坚

    2010-01-01

    使用环介导恒温扩增技术,基于莱姆病病原伯氏疏螺旋体的外膜蛋白A (OspA)基因,针对伯氏疏螺旋体不同的基因型设计特异性引物,对国内主要的莱姆病病原伯氏疏螺旋体的3个基因型进行分型鉴定.研究结果表明,设计的引物具有良好的特异性,可以对狭义伯氏疏螺旋体(Borrelia burgdorferi sensu strict)、嘎氏疏螺旋体(B.afzelii)和伽氏疏螺旋体(B.garinii)进行分型鉴定.伯氏疏螺旋体的分型鉴定可以对不同临床症状莱姆病患者的治疗和莱姆病的控制提供一定的依据.

  14. Horizontally Acquired Genes for Purine Salvage in Borrelia spp. Causing Relapsing Fever

    OpenAIRE

    Barbour, Alan G.; Putteet-Driver, Adrienne D.; Bunikis, Jonas

    2005-01-01

    Unlike Borrelia burgdorferi, the relapsing fever agent Borrelia hermsii and the related Borrelia miyamotoi had purA and purB genes of the purine salvage pathway. These were located among the rRNA genes. Phylogenetic analysis indicated that these genes had a different evolutionary history than those of orthologs in other spirochetes.

  15. 青海省互助、泽库和祁连县啮齿类动物伯氏疏螺旋体调查%Investigation of Borrelia burgdorferi in Glires in Huzhu, Zekog, and Qilian county, Qinghai

    Institute of Scientific and Technical Information of China (English)

    张琳; 石燕; 耿震; 侯学霞; 郝琴

    2015-01-01

    目的:调查确定青海省3县啮齿类标本中伯氏疏螺旋体带菌情况。方法从青海省互助、泽库和祁连县共采集啮齿类动物202只,采用巢式PCR方法检测其伯氏疏螺旋体的带菌率。结果采用巢式PCR方法检测啮齿类动物标本202份,阳性49份,伯氏疏螺旋体平均阳性率为24.26%。其中互助县标本共67份,阳性12份,阳性率为17.91%;泽库县标本共78份,阳性27份,阳性率为34.62%;祁连县标本共57只,阳性10份,阳性率为17.54%。3个县的啮齿类动物伯氏疏螺旋体阳性率差异有统计学意义(χ2=7.40,P<0.05)。结论青海省互助、泽库和祁连县啮齿类动物中均存在伯氏疏螺旋体感染,应进一步对当地的媒介和人群进行调查,从而为当地莱姆病的防控提供参考。%Objective To identify the Borrelia burgdorferi in Glires in Huzhu, Zekog, and Qilian county, Qinghai province. Methods A total of 202 Glires were collected from forest in the counties, Qinghai province. All the collected samples were examined for B. burgdorferi by nested PCR. Results In total of 202 samples, 49 samples were tested positive by 24.26%. Positive rate was 17.91%(12/67) in Huzhu county, 34.62%(27/78) in Zekog county and 17.54%(10/57) in Qilian county. There was significant difference with positive rate of B. burgdorferi in Glires among these three counties(χ2=7.40, P<0.05). Conclusion Our research confirmed the existence of B. burgdorferi in Glires in three counties in Qinghai province. We suggested that the further investigation on local vectors and human infections be conducted for prevention and control of Lyme disease.

  16. 莱姆病螺旋体感染对蜱的4个功能基因的转录影响%Effects of Borrelia burgdorferi infection on the transcription of four functional genes in the tick

    Institute of Scientific and Technical Information of China (English)

    吴银娟; 李国清; 仝彩玲; 曹杰; 周勇志; 周金林

    2011-01-01

    To explore the new strategy for preventing and controlling the tick and vector-bome disease, and to understand the molecular mechanism of interaction between pathogen and vector ticks, we studied the effect on the transcription of the four functional genes of tick after Borrelia burgdorfen infection. Total RNA was isolated at different time points from Haemaphysalis longicomis microinjected with B. burgdorferi suspension at different concentrations. After reverse transcription into cDNA, quantitative real-time fluorescence PCR was used to analyze the expression level of the four functional genes. The resulta showed that it had a significant effect on the transcription of the functional genes encoding cystatin, folliatatin-related protein and hemalin of the ticks by B. burgdorferi infection, despite the existence of dose-and time-dependent manners. On the fourth day, the expression of HLcyst-1 gene dramatically induced, but the expression of hemalin gene was inhibited.%为探索蜱及蜱传病的防控新策略,了解病原与媒介硬蜱相互作用的分子机制,本研究以莱姆病病原伯氏疏螺旋体感染后对长角血蜱4个功能基因的转录影响进行了研究.将不同浓度梯度的伯氏疏螺旋体显微注射到长角血蜱体内,分不同的时间段提取蜱的总RNA,反转录成cDNA后,用实时荧光定量PCR检测蜱基因的表达水平.结果显示:伯氏疏螺旋体感染对蜱半胱氨酸蛋白酶抑制剂1(HLcyst-1)、半胱氨酸蛋白酶抑制剂3(HLcyst-3)、卵泡抑素(FRP)、凝血酶抑制剂(Hemalin)4个功能基因的转录均产生了显著影响,但存在时间和剂量相关性,感染后第4天显著诱导了长角血蜱HLcyst-1基因的表达,抑制了Hemalin基因表达.

  17. 伯氏疏螺旋体分子生物学分型进展及其意义%Advance in molecular biologic typing of Borrelia burgdorferi and its significance

    Institute of Scientific and Technical Information of China (English)

    李建民; 曹务春

    2002-01-01

    @@ 姆病是一种蜱传全球性动物源性疾病,病原体由W.Burgdorfer首次分离并被命名为伯氏疏螺旋体.根据DNA和蛋白质分析,伯氏疏螺旋体可以分为10个基因型或基因群,即B.burgdorferi sensu stricto(B31T,标准株为B31;B.为Borrelia.的简写)、B.afzelii(VS461)、B.garinii(20047)、B.japonica(HO14)、B.andersonii(21123)、B.lusitaiae(PotiB2)、B.valaisiana(vs116)、B.tanukii(HK501)、B.turdae(Ya501)和基因群DN127[1~7].分子生物学分型包括表型分型和基因分型,评价不同分子生物学分型方法主要依据可分型性、可重复性、分辨率、易操作性、及结果易解释性.分子生物学分型方法较多,包括G+C mol%、DNA-DNA杂交、核糖体分型、PCR-RFLP、染色体DNA脉冲电泳、蛋白质电泳和多位点酶电泳、PCR-杂交、基因型特异性PCR及核酸序列聚类分析技术等.

  18. Frequency of antibodies to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burdgorferi in cattle from the northeastern region of the state of Pará, Brazil Freqüência de anticorpos para Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax e Borrelia burgdorferi em bovinos do nordeste do Estado do Pará, Brasil

    Directory of Open Access Journals (Sweden)

    Daniel S. Guedes Junior

    2008-06-01

    Full Text Available Babesiosis, anaplasmosis, and trypanosomosis are relevant diseases, potentially causing morbidity in cattle, leading to economic losses. Borreliosis is import as a potential zoonosis. The objective of this study was to determine, by indirect enzyme-linked immunosorbent assay (ELISA, the frequency of seropositive cattle to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burgdorferi in cattle from the Northeastern region of Pará, Brazil. Sera samples from 246 female adult cattle from municipalities of Castanhal and São Miguel do Guamá were used. Crude antigens ELISAs were used to detect antibodies to all agents, except to A. marginale, to which an indirect ELISA with recombinant major surface 1a protein (MSP1a antigen was used. Overall frequencies of seropositive animals were: B. bigemina - 99.2%; B. bovis - 98.8%; A. marginale - 68.3%; T. vivax - 93.1% and B. burgdorferi - 54.9%. The frequencies of seropositive cattle to B. bovis and B. bigemina suggest a high rate of transmission of these organisms by tick in the studied region, which can be classified as enzootically stable to these hemoprotozoans. The low frequency of seropositive cattle to A. marginale may be attributed to a lower sensitivity of the recombinant antigen ELISA utilized or a distinct rate of inoculation of this rickettsia by ticks, as compared with Babesia sp. transmission. The high frequency of seropositive cattle to T. vivax indicates that this hemoprotozoan is prevalent in herds from the Northeastern region of Pará. The rate of animal that showed homologues antibodies to B. burgdorferi indicates the presence of the tickborne spirochaetal agent in the cattle population in the studied region.A babesiose, a anaplasmose e a tripanossomose são enfermidades relevantes, potencialmente causadoras de morbidade em bovinos, levando a perdas econômicas. A borreliose assume importância como zoonose potencial. O objetivo desse estudo foi determinar

  19. The dynamic proteome of Lyme disease Borrelia.

    Science.gov (United States)

    Norris, Steven J

    2006-01-01

    The proteome of the spirochete bacterium Borrelia burgdorferi, the tick-borne agent of Lyme disease, has been characterized by two different approaches using mass spectrometry, providing a launching point for future studies on the dramatic changes in protein expression that occur during transmission of the bacterium between ticks and mammals. PMID:16563176

  20. The dynamic proteome of Lyme disease Borrelia

    OpenAIRE

    Norris, Steven J

    2006-01-01

    The proteome of the spirochete bacterium Borrelia burgdorferi, the tick-borne agent of Lyme disease, has been characterized by two different approaches using mass spectrometry, providing a launching point for future studies on the dramatic changes in protein expression that occur during transmission of the bacterium between ticks and mammals.

  1. A prospective study on the incidence of Borrelia burgdorferi sensu lato infection after a tick bite in Sweden and on the Åland Islands, Finland (2008-2009).

    Science.gov (United States)

    Wilhelmsson, Peter; Fryland, Linda; Lindblom, Pontus; Sjöwall, Johanna; Ahlm, Clas; Berglund, Johan; Haglund, Mats; Henningsson, Anna J; Nolskog, Peter; Nordberg, Marika; Nyberg, Clara; Ornstein, Katharina; Nyman, Dag; Ekerfelt, Christina; Forsberg, Pia; Lindgren, Per-Eric

    2016-02-01

    Lyme borreliosis (LB) is a common and increasing tick-borne disease in Europe. The risk of acquiring a Borrelia infection after a tick bite is not fully known. Therefore, we investigated the incidence of Borrelia infection after a bite by a Borrelia-infected tick and if the Borrelia load and/or the duration of tick-feeding influenced the risk of infection. During 2008-2009, ticks and blood samples were collected from 1546 tick-bitten persons from Sweden and the Åland Islands, Finland. Follow-up blood samples were taken 3 months after the tick bite. The duration of tick feeding was microscopically estimated and Borrelia was detected and quantified in ticks by real-time PCR. Anti-Borrelia antibodies were detected in sera using ELISA tests and immunoblot. Five percent (78/1546) of the study participants developed Borrelia infection (LB diagnosis and/or seroconversion) after a tick bite (45% bitten by Borrelia-infected ticks and 55% bitten by uninfected ticks). Of these, 33 developed LB (whereof 9 also seroconverted) while 45 participants seroconverted only. Experience of non-specific symptoms was more frequently reported by Borrelia-infected participants compared to uninfected participants. All who seroconverted removed "their" ticks significantly later than those who did not. The Borrelia load in the ticks did not explain the risk of seroconversion. Regional and sex differences in the Borrelia seroprevalence were found. The risk of developing a Borrelia infection after a bite by a Borrelia-infected tick is small but increases with the duration of tick feeding. PMID:26341726

  2. The function of histidine kinase Ⅱ(Hk2)in the life cycle of Borrelia burgdorferi%组氨酸激酶Ⅱ在伯氏疏螺旋体生活史中的功能

    Institute of Scientific and Technical Information of China (English)

    杨章女; 蒋宝贵; 杨晓峰; 徐海君; 曹务春

    2012-01-01

    目的 伯氏疏螺旋体组氨酸激酶Ⅱ(Hk2)和反应调节蛋白Rrp2共同组成一个二元信号系统,控制病原菌侵染哺乳动物所需致病因子的表达,但Hk2的生物学功能有待验证.方法 应用同源重组方法得到hk2敲除菌株(hk2-),并利用蜱-鼠动物模型对其生物学功能进行探索.结果 hk2-能够通过人工针刺正常感染小鼠,间接免疫荧光试验显示在吸血的幼蜱和若蜱中肠能够检测到hk2-,定量PCR结果显示hk2敲除并不影响病原菌在若蜱中肠的繁殖,但在蜱叮咬状态下只有50%的小鼠能够被hk2-感染.结论 Hk2不影响病原菌从鼠传播到幼蜱及随后在蜱体内的长期寄生,但hk2敲除降低了病原菌在自然条件下(蜱叮咬)感染小鼠的能力,研究结果暗示Hk2在病原菌生活史中的功能是协助病原菌高效地侵染哺乳动物.%Objective To validate the biological function of Hk2, which is reported to be in combination with the cognate response regulator Rrp2 to constitute a two component signal system in Borrelia burgdorferi that controls the key virulence factors required for pathogen invasion in mammals. Methods A hk2 knockout strain (hk2~) was generated through homologous recombination, with the biological function of Hk2 estimated by the mouse-tick animal model. Results The hk2~ knockout strain remained to be able to infect mouse by needle inoculation, and subsequently successfully detected in the midgut of fed-larvae and -nymph by IFA. The quantitative - PCR demonstrated that hk2 knockout had no influence on bacterial replication in the midgut of fed nymph. However, only 50% of tested mice were infected in hk2~ group by tick bite. Conclusion The hk2 knockout does not affect the pathogen acquisition from mouse by larvae ticks and subsequent persistent colonization in tick midgut, but it embarrasses the transmission from nymph to mouse under natural conditions by tick bite. The above data indicate that the Hk2 may effectively

  3. Determination of Novel Borrelia Genospecies in Swedish Ixodes ricinus Ticks

    OpenAIRE

    2002-01-01

    A total of 301 adult questing Ixodes ricinus ticks were collected at 15 different locations along the south and east coasts of Sweden to determine the Borrelia genospecies diversity. Thirty-two ticks (11%) were found to be positive by nested PCR with Borrelia burgdorferi sensu lato-specific primers. Species determination was based on partial sequencing of the 16S rRNA gene and the flagellin gene. Five different Borrelia species were found. The nucleotide sequence of the Borrelia DNA found in ...

  4. Purine Salvage Pathways among Borrelia Species▿

    Science.gov (United States)

    Pettersson, Jonas; Schrumpf, Merry E.; Raffel, Sandra J.; Porcella, Stephen F.; Guyard, Cyril; Lawrence, Kevin; Gherardini, Frank C.; Schwan, Tom G.

    2007-01-01

    Genome sequencing projects on two relapsing fever spirochetes, Borrelia hermsii and Borrelia turicatae, revealed differences in genes involved in purine metabolism and salvage compared to those in the Lyme disease spirochete Borrelia burgdorferi. The relapsing fever spirochetes contained six open reading frames that are absent from the B. burgdorferi genome. These genes included those for hypoxanthine-guanine phosphoribosyltransferase (hpt), adenylosuccinate synthase (purA), adenylosuccinate lyase (purB), auxiliary protein (nrdI), the ribonucleotide-diphosphate reductase alpha subunit (nrdE), and the ribonucleotide-diphosphate reductase beta subunit (nrdF). Southern blot assays with multiple Borrelia species and isolates confirmed the presence of these genes in the relapsing fever group of spirochetes but not in B. burgdorferi and related species. TaqMan real-time reverse transcription-PCR demonstrated that the chromosomal genes (hpt, purA, and purB) were transcribed in vitro and in mice. Phosphoribosyltransferase assays revealed that, in general, B. hermsii exhibited significantly higher activity than did the B. burgdorferi cell lysate, and enzymatic activity was observed with adenine, hypoxanthine, and guanine as substrates. B. burgdorferi showed low but detectable phosphoribosyltransferase activity with hypoxanthine even though the genome lacks a discernible ortholog to the hpt gene in the relapsing fever spirochetes. B. hermsii incorporated radiolabeled hypoxanthine into RNA and DNA to a much greater extent than did B. burgdorferi. This complete pathway for purine salvage in the relapsing fever spirochetes may contribute, in part, to these spirochetes achieving high cell densities in blood. PMID:17502392

  5. Whole-Genome Sequences of Two Borrelia afzelii and Two Borrelia garinii Lyme Disease Agent Isolates

    Energy Technology Data Exchange (ETDEWEB)

    Casjens, S.R.; Dunn, J.; Mongodin, E. F.; Qiu, W.-G.; Luft, B. J.; Fraser-Liggett, C. M.; Schutzer, S. E.

    2011-12-01

    Human Lyme disease is commonly caused by several species of spirochetes in the Borrelia genus. In Eurasia these species are largely Borrelia afzelii, B. garinii, B. burgdorferi, and B. bavariensis sp. nov. Whole-genome sequencing is an excellent tool for investigating and understanding the influence of bacterial diversity on the pathogenesis and etiology of Lyme disease. We report here the whole-genome sequences of four isolates from two of the Borrelia species that cause human Lyme disease, B. afzelii isolates ACA-1 and PKo and B. garinii isolates PBr and Far04.

  6. Genetic and antigenic characterization of Borrelia coriaceae, putative agent of epizootic bovine abortion.

    OpenAIRE

    LeFebvre, R B; Perng, G C

    1989-01-01

    Borrelia coriaceae was characterized genetically and antigenically by utilizing the following techniques: restriction endonuclease analysis, Southern blotting and genomic hybridization, pulsed-field electrophoresis, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting. The B. coriaceae genome revealed unique and characteristic banding patterns both by agarose gel electrophoresis and by hybridization when compared with several Borrelia burgdorferi isolates. Pulsed-fiel...

  7. Antinuclear antibodies are not increased in the early phase of Borrelia infection.

    OpenAIRE

    Spiewak, R.W.; Stojek, NM; Chmielewska-Badora, J.

    2004-01-01

    In the literature, there are case reports suggesting that Borrelia burgdorferi infection may induce autoimmune diseases dependent on antinuclear antibodies (ANA). The present study was undertaken in order to verify this possibility in a prospective manner. The study group comprised 78 consecutive patients (51 women and 27 men, median age 41.5 years) referred to our Department for the serologic diagnosis of Borrelia infection. The patients' sera were tested for Borrelia-specific IgM and IgG (R...

  8. Phagosomal TLR signaling upon Borrelia burgdorferi infection

    Science.gov (United States)

    Cervantes, Jorge L.; Hawley, Kelly L.; Benjamin, Sarah J.; Weinerman, Bennett; Luu, Stephanie M.; Salazar, Juan C.

    2014-01-01

    Internalization and degradation of live Bb within phagosomal compartments of monocytes, macrophages and dendritic cells (DCs), allows for the release of lipoproteins, nucleic acids and other microbial products, triggering a broad and robust inflammatory response. Toll-like receptors (TLRs) are key players in the recognition of spirochetal ligands from whole viable organisms (i.e., vita-PAMPs). Herein we will review the role of endosomal TLRs in the response to the Lyme disease spirochete. PMID:24904837

  9. Borrelia infection and risk of non-Hodgkin lymphoma

    DEFF Research Database (Denmark)

    Schollkopf, C.; Melbye, M.; Munksgaard, L.;

    2008-01-01

    population controls. History of tick bite or Borrelia infection was ascertained through structured telephone interviews and through enzyme-linked immunosorbent assay serum analyses for antibodies against B. burgdorferi in a subset of 1579 patients and 1358 controls. Statistical associations with risk of NHL......, including histologic subtypes, were assessed by logistic regression. Overall risk of NHL was not associated with self-reported history of tick bite (odds ratio [OR] = 1.0; 95% confidence interval: 0.9-1.1), Borrelia infection (OR = 1.3 [0.96-1.8]) or the presence of anti-Borrelia antibodies (OR = 1.3 [0...

  10. Borrelia miyamotoi-Associated Neuroborreliosis in Immunocompromised Person.

    Science.gov (United States)

    Boden, Katharina; Lobenstein, Sabine; Hermann, Beate; Margos, Gabriele; Fingerle, Volker

    2016-09-01

    Borrelia miyamotoi is a newly recognized human pathogen in the relapsing fever group of spirochetes. We investigated a case of B. miyamotoi infection of the central nervous system resembling B. burgdorferi-induced Lyme neuroborreliosis and determined that this emergent agent of central nervous system infection can be diagnosed with existing methods. PMID:27533748

  11. Associations of passerine birds, rabbits, and ticks with Borrelia miyamotoi and Borrelia andersonii in Michigan, U.S.A.

    Directory of Open Access Journals (Sweden)

    Hamer Sarah A

    2012-10-01

    Full Text Available Abstract Background Wild birds contribute to maintenance and dissemination of vectors and microbes, including those that impact human, domestic animal, and wildlife health. Here we elucidate roles of wild passerine birds, eastern cottontail rabbits (Sylvilagus floridanus, and Ixodes dentatus ticks in enzootic cycles of two spirochetes, Borrelia miyamotoi and B. andersonii in a region of Michigan where the zoonotic pathogen B. burgdorferi co-circulates. Methods Over a four-year period, wild birds (n = 19,631 and rabbits (n = 20 were inspected for tick presence and ear tissue was obtained from rabbits. Samples were tested for Borrelia spirochetes using nested PCR of the 16S-23S rRNA intergenic spacer region (IGS and bidirectional DNA sequencing. Natural xenodiagnosis was used to implicate wildlife reservoirs. Results Ixodes dentatus, a tick that specializes on birds and rabbits and rarely bites humans, was the most common tick found, comprising 86.5% of the 12,432 ticks collected in the study. The relapsing fever group spirochete B. miyamotoi was documented for the first time in ticks removed from wild birds (0.7% minimum infection prevalence; MIP, in I. dentatus, and included two IGS strains. The majority of B. miyamotoi-positive ticks were removed from Northern Cardinals (Cardinalis cardinalis. Borrelia andersonii infected ticks removed from birds (1.6% MIP, ticks removed from rabbits (5.3% MIP, and rabbit ear biopsies (5% comprised twelve novel IGS strains. Six species of wild birds were implicated as reservoirs for B. andersonii. Frequency of I. dentatus larval and nymphal co-feeding on birds was ten times greater than expected by chance. The relatively well-studied ecology of I. scapularis and the Lyme disease pathogen provides a context for understanding how the phenology of bird ticks may impact B. miyamotoi and B. andersonii prevalence and host associations. Conclusions Given the current invasion of I. scapularis, a human biting species

  12. Genotypic variation and mixtures of Lyme Borrelia in Ixodes ticks from North America and Europe.

    Directory of Open Access Journals (Sweden)

    Chris D Crowder

    Full Text Available BACKGROUND: Lyme disease, caused by various species of Borrelia, is transmitted by Ixodes ticks in North America and Europe. Studies have shown the genotype of Borrelia burgdorferi sensu stricto (s.s. or the species of B. burgdorferi sensu lato (s.l. affects the ability of the bacteria to cause local or disseminated infection in humans. METHODOLOGY/PRINCIPAL FINDINGS: We used a multilocus PCR electrospray mass spectrometry assay to determine the species and genotype Borrelia from ticks collected in New York, Connecticut, Indiana, Southern Germany, and California and characterized isolates from parts of the United States and Europe. These analyses identified 53 distinct genotypes of B. burgdorferi sensu stricto with higher resolution than ospC typing. Genotypes of other members of the B. burgdorferi sensu lato complex were also identified and genotyped including B. afzelii, B. garinii, B. lusitaniae, B. spielmanii, and B. valaisiana. While each site in North America had genotypes unique to that location, we found genotypes shared between individual regions and two genotypes found across the United States. Significant B. burgdorferi s.s. genotypic diversity was observed between North America and Europe: only 6.6% of US genotypes (3 of 45 were found in Europe and 27% of the European genotypes (3 of 11 were observed in the US. Interestingly, 39% of adult Ixodes scapularis ticks from North America were infected with more than one genotype of B. burgdorferi s.s. and 22.2% of Ixodes ricinus ticks from Germany were infected with more than one genotype of B. burgdorferi s.l. CONCLUSIONS/SIGNIFICANCE: The presence of multiple Borrelia genotypes in ticks increases the probability that a person will be infected with more than one genotype of B. burgdorferi, potentially increasing the risks of disseminated Lyme disease. Our study indicates that the genotypic diversity of Borrelia in ticks in both North America and Europe is higher then previously reported

  13. Laimo boreliozės paplitimo Lietuvoje ir ligos sukėlėjo Borrelia burgdorferi s. l. ospA geno sekų analizė

    OpenAIRE

    Juodišiūtė, Indrė

    2014-01-01

    Laimo boreliozė (LB) yra labiausiai paplitusi erkių platinama liga. Ligos sukėlėjus perneša Ixodes rūšies erkės. Didelį borelijų paplitimą lemia platus rezervuarinių šeimininkų ratas. Šį susirgimą gali sukelti kelios B. burgdorferi s. l. komplekso bakterijų rūšys: B. afzelii, B. garinii, B. burgdorferi sensu stricto. Šiame darbe buvo tirtos 134 Ixodes ricinus erkės nuo 8 elninių gyvūnų (4 stirnų ir 4 elnių) ir 73 erkės nuo žolės (elninių teritorijoje). Naudojant dauginės polimerazės gra...

  14. Differential associations of Borrelia species with European badgers (Meles meles) and raccoon dogs (Nyctereutes procyonoides) in western Poland.

    Science.gov (United States)

    Wodecka, Beata; Michalik, Jerzy; Lane, Robert S; Nowak-Chmura, Magdalena; Wierzbicka, Anna

    2016-07-01

    European badgers and raccoon dogs and their associated ticks and lice were assayed for the presence of Lyme borreliosis and relapsing fever-group spirochete DNA in western Poland. Analyses of blood, ear-biopsy and liver samples revealed that 25% of 28 raccoon dogs and 12% of 34 badgers were PCR positive for borreliae. Borrelia garinii was the dominant species in raccoon dogs (62.5%), followed by B. afzelii (25%) and B. valaisiana (12.5%). PCR-positive badgers were infected only with B. afzelii. A total of 351 attached ticks was recovered from 23 (82%) of the raccoon dogs and 13 (38%) of the badgers. Using a nested PCR targeting the ITS2 fragments of Ixodes DNA, four Ixodes species were identified: I. ricinus, I. canisuga, I. hexagonus, and one provisionally named I. cf. kaiseri. Ixodes canisuga and I. ricinus prevailed on both host species. The highest infection prevalence was detected in I. ricinus, followed by I. canisuga and I. cf. kaiseri. Borrelia garinii and B. afzelii accounted for 61.6% and 30.1% of the infections detected in all PCR-positive ticks, respectively. Four other Borrelia species (B. burgdorferi sensu stricto, B. valaisiana, B. lusitaniae and B. miyamotoi) were detected only in I. ricinus from raccoon dogs. Moreover, Borrelia DNA, mostly B. garinii, was detected in 57 (81.4%) of 70 Trichodectes melis lice derived from 12 badgers. The detection of B. afzelii in one-half of PCR-positive biopsies reconfirms previous associations of this species with mammalian hosts, whereas the high prevalence of B. garinii in feeding lice and I. ricinus ticks (including larvae) demonstrates that both carnivores serve as hosts for B. garinii. The lack of B. garinii DNA in the tissues of badgers versus its prevalence in raccoon-dog biopsies, however, incriminates only the latter carnivore as a potential reservoir host. PMID:27263838

  15. Differential Role of Passerine Birds in Distribution of Borrelia Spirochetes, Based on Data from Ticks Collected from Birds during the Postbreeding Migration Period in Central Europe▿

    OpenAIRE

    Dubska, Lenka; Literak, Ivan; Kocianova, Elena; Taragelova, Veronika; Sychra, Oldrich

    2008-01-01

    Borrelia spirochetes in bird-feeding ticks were studied in the Czech Republic. During the postbreeding period (July to September 2005), 1,080 passerine birds infested by 2,240 Ixodes ricinus subadult ticks were examined. Borrelia garinii was detected in 22.2% of the ticks, Borrelia valaisiana was detected in 12.8% of the ticks, Borrelia afzelii was detected in 1.6% of the ticks, and Borrelia burgdorferi sensu stricto was detected in 0.3% of the ticks. After analysis of infections in which the...

  16. Structural Requirements for Glycosaminoglycan Recognition by the Lyme Disease Spirochete, Borrelia burgdorferi†

    OpenAIRE

    Leong, John M.; Robbins, Douglas; Rosenfeld, Louis; Lahiri, Biswajit; Parveen, Nikhat

    1998-01-01

    Borrelia burgdorferi, the Lyme disease agent, binds glycosaminoglycans (GAGs) such as heparin, heparan sulfate, and dermatan sulfate. Heparin or heparan sulfate fractions separated by size or charge were tested for their ability to inhibit attachment of B. burgdorferi to Vero cells. GAG chains of increasing length and/or charge showed increasing inhibitory potency, and detectable heparin inhibition of bacterial binding required a minimum of 16 residues. The ability of a given heparin fraction...

  17. How do Lyme Borrelia Organisms Cause Disease? The Quest for Virulence Determinants #

    OpenAIRE

    Norris, Steven J

    2012-01-01

    Lyme disease Borrelia are invasive, nontoxigenic, persistent pathogens, and little is known about their mechanisms of pathogenesis. In our laboratory, a signature-tagged mutagenesis (STM) library of over 4,000 Borrelia burgdorferi transposon mutants has been constructed and is being screened for infectivity in mice. In this manner, a global view of the virulence determinants (factors required for full infectivity) is being developed. Additionally, the mechanisms of immune evasion involving th...

  18. Transposon mutagenesis as an approach to improved understanding of Borrelia pathogenesis and biology

    OpenAIRE

    Lin, Tao; Troy, Erin B.; Linden T. Hu; Gao, Lihui; Norris, Steven J

    2014-01-01

    Transposon insertion provides a method for near-random mutation of bacterial genomes, and has been utilized extensively for the study of bacterial pathogenesis and biology. This approach is particularly useful for organisms that are relatively refractory to genetic manipulation, including Lyme disease Borrelia. In this review, progress to date in the application of transposon mutagenesis to the study of Borrelia burgdorferi is reported. An effective Himar1-based transposon vector has been dev...

  19. Transposon mutagenesis as an approach to improved understanding of Borrelia pathogenesis and biology

    OpenAIRE

    Tao eLin; Troy, Erin B.; Linden T. Hu; Lihui eGao; Norris, Steven J

    2014-01-01

    Transposon insertion provides a method for near-random mutation of bacterial genomes, and has been utilized extensively for the study of bacterial pathogenesis and biology. This approach is particularly useful for organisms that are relatively refractory to genetic manipulation, including Lyme disease Borrelia. In this review, progress to date in the application of transposon mutagenesis to the study of Borrelia burgdorferi is reported. An effective Himar1-based transposon vector has been ...

  20. MOLECULAR CLONING AND EXPRESSION IN BORRELIA BURGDORFERI P39 GENE E. COLI%莱姆病螺旋体P39蛋白的基因克隆及在大肠杆菌中的高效表达

    Institute of Scientific and Technical Information of China (English)

    佟玉品; 冯方波; 毕胜利; 江永珍; 鲁健

    2000-01-01

    目的克隆并表达我国莱姆病螺旋体分离株BmpA即P39基因,为制备莱姆病螺旋体基因工程重组抗原以用于我国莱姆病的基础与临床诊断研究奠定基础。方法应用PCR方法及基因重组技术对我国莱姆病螺旋体分离株P39蛋白进行全基因克隆,并在此基础上去掉信号肽序列,将P39基因插入原核表达载体LKB2,在大肠杆菌BL21(DE3)中进行诱导表达。表达产物以SDS-PAGE电泳、Western blot、薄层扫描分析。结果成功地克隆了P39基因,序列分析显示,P39基因全长1020bp,其核苷酸序列及氨基酸序列同源性与国外分离株均较高,与Sh-2-82株皆为99%,P39基因在大肠杆菌中以天然蛋白形式得到高效表达,扫描分析其分子量为37kDa,单株表达量占菌体总蛋白的58%,Western blot实验表明其可与莱姆病患者血清发生特异性反应。结论成功克隆了莱姆病螺旋体国内分离株P39蛋白基因并且在原核系统中得到高效表达。重组P39蛋白具有较好的免疫活性,可作为莱姆病新型诊断试剂理想的候选抗原。%Aim Molecular cloning and expressing BmpA(P39) gene from a Borrelia burgdorferi strain isolated from China for preparing recombinant antigen to the basic and chinical diagnostic study on Lyme disease in China. Method PCR and gene recombination technique were used to clone the whole P39 gene from a strain BT01. The P39 gene was then inserted into a expression vector LKB2 without its signal peptide and expressed in E. coli. The expressed protein was identified by SDS -PAGE,Western blot, and scanning analysis. Results Sequence analysis showed the P39 gene was 1020 base pairs with high homology to foreign isolates. Especially it had 99% nucleotide and amino acid identity with the Sh-2-82 strain. The recombinant plasmid BT-p39 expressed the protein with high level and a native form in the host cell BL21 (DE3),the protein was about 37KDa,and it accounted

  1. Virulence of recurrent infestations with Borrelia-infected ticks in a Borrelia-amplifying bird

    Science.gov (United States)

    Heylen, Dieter J. A.; Müller, Wendt; Vermeulen, Anke; Sprong, Hein; Matthysen, Erik

    2015-11-01

    Lyme disease cases caused by Borrelia burgdorferi s.l. bacteria is increasing steadily in Europe, in part due to the expansion of the vector, Ixodes ricinus. Wild reservoir hosts are typically recurrently infested. Understanding the impact of these cumulative parasite exposures on the host’s health is, therefore, central to predict the distribution of tick populations and their pathogens. Here, we have experimentally investigated the symptoms of disease caused by recurrent infestations in a common songbird (Parus major). Birds were exposed three times in succession to ticks collected in a Borrelia endemic area. Health and immune measures were analyzed in order to investigate changes in response to tick infestation and Borrelia infection rate. Nitric oxide levels increased with the Borrelia infection rate, but this effect was increasingly counteracted by mounting tick infestation rates. Tick infestations equally reduced haematocrit during each cycle. But birds overcompensated in their response to tick feeding, having higher haematocrit values during tick-free periods depending on the number of ticks they had been previously exposed to. Body condition showed a similar overshooting response in function of the severity of the Borrelia infection. The observed overcompensation increases the bird’s energetic needs, which may result in an increase in transmission events.

  2. Rats, the primary reservoir hosts of Borrelia burgdorferi, in six representative provinces ,China%中国6省莱姆病螺旋体主要宿主动物鼠的初步调查

    Institute of Scientific and Technical Information of China (English)

    侯学霞; 耿震; 郝琴; 张洋; 万康林

    2010-01-01

    目的 了解中国吉林、山西、甘肃、青海、贵州、湖南6省林区莱姆病螺旋体宿主动物鼠的感染情况.方法 在每个省各选取两个采样点进行捕鼠,采用病原分离培养和巢式PCR方法对野鼠的脾脏、肾脏和膀胱进行了病原学检测.并通过基因测序方法确定基因型.结果 从贵州黑线姬鼠中分离到了2株莱姆病螺旋体;从5省(贵州未检测到)野鼠的脾脏和/或肾脏中检查到了莱姆病螺旋体的特异片段,其中青海黄南(28.85%)和湖南石门(19.6%)两地标本的PCR阳性率较高,各地区阳性率的差异有统计学意义.通过序列同源性分析确定吉林、青海、甘肃和山西的基因型均为Borrelia garinii.湖南的基因型为Borrelia valaisiana.结论 本次调查表明各地宿主动物鼠的感染状况不同,各地应根据具体情况进一步扩大调查以明确当地的主要宿主动物种类及携带病原体情况.

  3. Longitudinal surveillance of the tick Ixodes ricinus for borreliae

    Czech Academy of Sciences Publication Activity Database

    Hubálek, Zdeněk; Halouzka, Jiří; Juřicová, Zina

    2003-01-01

    Roč. 17, č. 1 (2003), s. 46-51. ISSN 0269-283X R&D Projects: GA ČR GA206/00/1204; GA AV ČR IBS6093007 Institutional research plan: CEZ:AV0Z6093917 Keywords : Borrelia burgdorferi * Ixodes ricinus * Lyme disease Subject RIV: FN - Epidemiology, Contagious Disease s ; Clinical Immunology Impact factor: 1.040, year: 2003

  4. Borreliae in Ixodes ricinus ticks feeding on humans

    Czech Academy of Sciences Publication Activity Database

    Hubálek, Zdeněk; Halouzka, Jiří; Juřicová, Zina

    2004-01-01

    Roč. 18, č. 3 (2004), s. 228-231. ISSN 0269-283X R&D Projects: GA ČR GA206/03/0726; GA AV ČR IBS6093007 Institutional research plan: CEZ:AV0Z6093917 Keywords : Borrelia burgdorferi * Ixodes ricinus * Lyme disease Subject RIV: FN - Epidemiology, Contagious Disease s ; Clinical Immunology Impact factor: 1.405, year: 2004

  5. Associations of passerine birds, rabbits, and ticks with Borrelia miyamotoi and Borrelia andersonii in Michigan, U.S.A.

    OpenAIRE

    Hamer Sarah A; Hickling Graham J; Keith Rich; Sidge Jennifer L; Walker Edward D; Tsao Jean I

    2012-01-01

    Abstract Background Wild birds contribute to maintenance and dissemination of vectors and microbes, including those that impact human, domestic animal, and wildlife health. Here we elucidate roles of wild passerine birds, eastern cottontail rabbits (Sylvilagus floridanus), and Ixodes dentatus ticks in enzootic cycles of two spirochetes, Borrelia miyamotoi and B. andersonii in a region of Michigan where the zoonotic pathogen B. burgdorferi co-circulates. Methods Over a four-year period, wild b...

  6. 从桂黔交界地区中华硬蜱中分离到莱姆病螺旋体%Borrelia burgdorferi isolated from Ixodes sinensis in the border areas between Guangxi and Guizhou in China

    Institute of Scientific and Technical Information of China (English)

    李永学; 王昭孝; 曾霞; 王树声; 余春; 周敬祝

    2010-01-01

    目的 从桂黔交界地区捕获的蜱中分离莱姆病螺旋体,以探查广西壮族自治区(广西区)和贵州省莱姆病的传播媒介.方法 用BSK-Ⅱ培养基从蜱中分离莱姆病螺旋体.用PCR扩增分离菌株的5S~23S rRNA基因间隔区,将PCR扩增产物克隆后测序,通过将测序结果与基因库中莱姆病螺旋体菌株的5S~23S rRNA基因间隔区进行同源性比较和分析,从而鉴定分离株的基因型.结果 从捕获的中华硬蜱中分离出1株莱姆病螺旋体(命名为QLT1).分离株经鉴定为Borrelia valaisiana基因型莱姆病螺旋体.结论 中华硬蜱很有可能是广西区和贵州省莱姆病的传播媒介.

  7. 我国天津地区莱姆病螺旋体宿主动物的初步调查%Preliminary Investigation on Reservoir Hosts of Borrelia Burgdorferi in the Area of Tianjin

    Institute of Scientific and Technical Information of China (English)

    郝琴; 刘义; 耿震; 陈继永; 侯学霞; 宋春英; 陈建; 丁健青; 于风泰; 贾艳盒; 赵志武; 万康林

    2007-01-01

    目的 了解天津地区莱姆病螺旋体的宿主动物的感染情况.方法 采用间接免疫荧光抗体法(IFA)对野鼠、野兔、牛、羊、狗进行了莱姆病螺旋体感染的血清学检测;用病原分离培养方法和 PCR 方法对野鼠进行了病原学检测.并通过基因测序方法确定基因型.结果 野鼠的血清莱姆病感染阳性率最高为 32.00%(32/100);并从大林姬鼠、社鼠、小林姬鼠的脾脏和/或肾脏中检查到莱姆病螺旋体的特异片断.通过序同源性分析确定为Borrelia garinii 基因型.结论 首次从病原学证实天津林区鼠类为莱姆病螺旋体的重要储存宿主.

  8. Insights into the complex regulation of rpoS in Borrelia burgodorferi

    Science.gov (United States)

    The coordinated regulation of gene expression is required for the transmission and survival of Borrelia burgdorferi in different hosts, and the sigma factor RpoS (sigmaS), as regulated by RpoN (sigma54), has been shown to regulate key proteins in these processes. We show that rpoS has one sigma54-d...

  9. Antinuclear antibodies are not increased in the early phase of Borrelia infection.

    NARCIS (Netherlands)

    Spiewak, R.W.; Stojek, NM; Chmielewska-Badora, J

    2004-01-01

    In the literature, there are case reports suggesting that Borrelia burgdorferi infection may induce autoimmune diseases dependent on antinuclear antibodies (ANA). The present study was undertaken in order to verify this possibility in a prospective manner. The study group comprised 78 consecutive pa

  10. Tick sialostatins L and L2 differentially influence dendritic cell responses to Borrelia spirochetes

    Czech Academy of Sciences Publication Activity Database

    Lieskovská, Jaroslava; Páleníková, Jana; Langhansová, Helena; Chagas, A. C.; Calvo, E.; Kotsyfakis, Michalis; Kopecký, Jan

    2015-01-01

    Roč. 8, MAY 15 2015 (2015), s. 275. ISSN 1756-3305 R&D Projects: GA ČR GAP302/12/2208 Institutional support: RVO:60077344 Keywords : Dendritic cells * Borrelia burgdorferi * Tick cystatin * Signalling Subject RIV: EC - Immunology Impact factor: 3.430, year: 2014

  11. Prevalence of borreliae in ixodid ticks from a floodplain forest ecosystem

    Czech Academy of Sciences Publication Activity Database

    Hubálek, Zdeněk; Stünzner, D.; Halouzka, Jiří; Sixl, W.; Wendelin, I.; Juřicová, Zina; Sanogo, Yibayiri Osée

    2003-01-01

    Roč. 115, 3-4 (2003), s. 121-124. ISSN 0043-5325 R&D Projects: GA MŠk ME 229; GA ČR GA206/00/1204 Institutional research plan: CEZ:AV0Z6093917 Keywords : Lyme disease * Borrelia burgdorferi * floodplain forest Subject RIV: EE - Microbiology, Virology Impact factor: 0.774, year: 2003

  12. Detection and genotyping of Borrelia burgdorferi sensu lato in Pingxiang of Guangxi Zhuang Autonomous Region, China%广西凭祥地区莱姆病螺旋体检测和基因分型研究

    Institute of Scientific and Technical Information of China (English)

    韩华; 万道正; 张晓龙; 梁中平; 郭天宇; 褚宸一; 赵荣涛; 石华; 宋宏彬

    2013-01-01

    目的 调查广西凭祥地区莱姆病感染情况和基因型别.方法 2011年7月从广西凭祥地区采集蜱、啮齿动物和野鸟标本,分别采取煮沸法和Qiagen试剂盒提取蜱、啮齿动物脾脏以及鸟脾脏中莱姆病螺旋体基因组DNA;采用巢式PCR扩增莱姆病螺旋体5S~23S rRNA基因间隔区;对PCR扩增产物进行测序,并将序列结果与GenBank中莱姆病螺旋体5S~23SrRNA基因问隔区序列比对分析.结果 从3份啮齿动物标本中检测到莱姆病螺旋体5S~23SrRNA基因间隔区片段,啮齿动物的感染率为5.66%(3/53).其中一个序列与GenBank中Borrelia valaisiana基因型莱姆病螺旋体(序列号:HM100125.1,AB091455.1,AB091454.1,AB091453.1)同源性为100%;蜱和鸟标本中未检测到莱姆病螺旋体.结论 广西凭祥地区啮齿动物中存在莱姆病螺旋体B.valaisiana基因型感染.

  13. Preparation and preliminary identification of monoclonal antibody against Borrelia burgdorferi in China%中国莱姆病螺旋体特异性单克隆抗体的制备及初步鉴定

    Institute of Scientific and Technical Information of China (English)

    郝琴; 曹伯良; 侯学霞; 耿震; 万康林

    2008-01-01

    目的 制备特异的莱姆病螺旋体单克隆抗体,为我国莱姆病的诊断和莱姆病螺旋体菌株鉴定提供基础.方法 以中国莱姆病螺旋体伽氏疏螺旋体(Borrelia garinii)的代表菌株PD91的全菌蛋白为抗原,免疫BALB/c小鼠,取脾细胞与骨髓瘤细胞SP2/0融合,用间接酶联免疫吸附试验(ELISA)和蛋白免疫印迹方法(WB)筛选,并经过2次或3次克隆,以获得单克隆抗体.结果 共制备出10株单克隆抗体,经鉴定为3种,分别针对中国莱姆病螺旋体的外膜蛋白OspA(4株)、OspB(3株)和OspC(3株).结论 成功制备出3种抗莱姆病螺旋体不同蛋白的单克隆抗体, 可用于我国莱姆病的病原诊断和莱姆病螺旋体菌株鉴定.

  14. Molecular identification of Borrelia genus in questing hard ticks from Portugal: Phylogenetic characterization of two novel Relapsing Fever-like Borrelia sp.

    Science.gov (United States)

    Nunes, Mónica; Parreira, Ricardo; Maia, Carla; Lopes, Nádia; Fingerle, Volker; Vieira, M Luísa

    2016-06-01

    In the last decades, several studies have reported pathogenic species of Borrelia related to those that cause Tick-borne Relapsing Fever (RF), but unexpectedly suggesting their transmission by hard ticks, known vectors of Borrelia burgdorferi sensu lato (B. burgdorferi s.l.) species, rather than by soft ticks. This study was designed to update the presence of B. burgdorferi s.l. species in ticks from several districts of mainland Portugal, where Ixodes ricinus had been previously described. Ticks (a total of 2915 specimens) were collected in seven districts throughout the country, and analyzed using molecular methods. Three nested-PCR protocols, targeting the flagellin gene (flaB), the intergenic spacer region (IGS) located between 5S and 23S rRNA, and the glpQ gene, and a conventional PCR targeting the 16S rRNA, were used for Borrelia DNA detection. Borrelia DNA was detected in 3% of the ticks from Braga, Vila Real, Lisboa, Setúbal, Évora and Faro districts. The obtained amplicons were sequenced and analyzed by BLASTn, and 15/63 (24%) matched with homologous sequences from Borrelia lusitaniae and 15/63 (24%) with B. garinii, being these the most prevalent species. DNA from B. burgdorferi sensu stricto (s.s.), B. valaisiana and B. afzelii were detected in 7/63 (11%), 6/63 (10%), and 2/63 (3%) of the specimens, respectively. Unexpectedly, DNA sequence (flaB) analysis from eight (13%) samples, two from Rhipicephalus sanguineus and six from Haemaphysalis punctata tick species, revealed high homology with RF-like Borrelia. Phylogenetic analyses obtained from three genetic markers (16S rRNA, flaB, and glpQ) confirmed their congruent inclusion in a strongly supported RF cluster, where they segregated in two subgroups which differ from the other Relapsing Fever species. Therefore, the results confirm the circulation of multiple species of B. burgdorferi s.l. over a wide geographic range, covering most of the Portuguese mainland territory. Surprisingly, the obtained data

  15. Detection of specific antibody IgM of Lyme disease by ELISA using recombinant outer surface protein C of borrelia burgdorferi%用基因工程抗原建立ELISA检测莱姆病特异IgM抗体

    Institute of Scientific and Technical Information of China (English)

    贾月萍; 周国萍; 曾丽苹; 高峰

    2001-01-01

    目的利用伯氏疏螺旋体基因工程抗原外膜蛋白C(OspC)建立间接ELISA,检测莱姆病特异性抗体IgM。方法基因工程抗原OspC的包被浓度和酶标抗μ链单抗所用浓度及血清稀释倍数,均由方阵滴定法确定,并进行精密度、特异性试验、阻断试验和干扰试验。结果 OspC最佳浓度为150 μg/L,批内平均变异系数4.6%,批间平均变异系数14.2%,用ELISA测定临床已确诊莱姆病33例,57例正常体检者,同时与进口ELISA试剂盒比较,两方法符合率97.8%。结论该方法特异性强、敏感性高、实验结果可靠,是莱姆病早期诊断的好方法。%Objective To detect specific IgM of Lyme disease indirect ELISA using recombinant outer surface protein C(OpsC)of Borrelia burgdorferi in vitro was established. Methods Coated concentration of recombinant OspC and dilution multiple of serum anol concentration of enzyme secondary antibody were determined by block,and degree of percision.specificity interference and interruption test were performed. Results Best concentration of OspC was 150 μg/L.within-run CV was 6.3% between-run CV was 11.8%.Confimred 33 Lyme patients in clinic and 57 controls were examined meanwhile by this method and import ELISA kit,coincidena rate was 97.8%.Conclusion This ELISA using recombinant OspC was a good method for early diagnostic of Lyme disease.

  16. Vector competence of the blacklegged tick, Ixodes scapularis, for the recently recognized Lyme borreliosis spirochete Candidatus Borrelia mayonii.

    Science.gov (United States)

    Dolan, Marc C; Hojgaard, Andrias; Hoxmeier, J Charles; Replogle, Adam J; Respicio-Kingry, Laurel B; Sexton, Christopher; Williams, Martin A; Pritt, Bobbi S; Schriefer, Martin E; Eisen, Lars

    2016-07-01

    A novel species within the Borrelia burgdorferi sensu lato complex, provisionally named Borrelia mayonii, was recently found to be associated with Lyme borreliosis in the Upper Midwest of the United States. Moreover, B. mayonii was detected from host-seeking Ixodes scapularis, the primary vector of B. burgdorferi sensu stricto in the eastern United States. We therefore conducted a study to confirm the experimental vector competence of I. scapularis for B. mayonii (strain MN14-1420), using colony ticks originating from adults collected in Connecticut and CD-1 white mice. Larvae fed on mice 10 weeks after needle-inoculation with B. mayonii acquired spirochetes and maintained infection through the nymphal stage at an average rate of 12.9%. In a transmission experiment, 40% of naïve mice exposed to a single infected nymph developed viable infections, as compared with 87% of mice fed upon by 2-3 infected nymphs. Transmission of B. mayonii by one or more feeding infected nymphs was uncommon up to 48h after attachment (one of six mice developed viable infection) but occurred frequently when nymphs were allowed to remain attached for 72-96h or feed to completion (11 of 16 mice developed viable infection). Mice infected via tick bite maintained viable infection with B. mayonii, as determined by ear biopsy culture, for at least 28 weeks. Our results demonstrate that I. scapularis is capable of serving as a vector of B. mayonii. This finding, together with data showing that field-collected I. scapularis are infected with B. mayonii, indicate that I. scapularis likely is a primary vector to humans of this recently recognized Lyme borreliosis spirochete. PMID:26922324

  17. Cooperation of Doxycycline with Phytochemicals and Micronutrients Against Active and Persistent Forms of Borrelia sp.

    Science.gov (United States)

    Goc, Anna; Niedzwiecki, Alexandra; Rath, Matthias

    2016-01-01

    Phytochemicals and micronutrients represent a growing theme in antimicrobial defense; however, little is known about their anti-borreliae effects of reciprocal cooperation with antibiotics. A better understanding of this aspect could advance our knowledge and help improve the efficacy of current approaches towards Borrelia sp. In this study, phytochemicals and micronutrients such as baicalein, luteolin, 10-HAD, iodine, rosmarinic acid, and monolaurin, as well as, vitamins D3 and C were tested in a combinations with doxycycline for their in vitro effectiveness against vegetative (spirochetes) and latent (rounded bodies, biofilm) forms of Borrelia burgdorferi and Borrelia garinii. Anti-borreliae effects were evaluated according to checkerboard assays and supported by statistical analysis. The results showed that combination of doxycycline with flavones such as baicalein and luteolin exhibited additive effects against all morphological forms of studied Borrelia sp. Doxycycline combined with iodine demonstrated additive effects against spirochetes and biofilm, whereas with fatty acids such as monolaurin and 10-HAD it produced FICIs of indifference. Additive anti-spirochetal effects were also observed when doxycycline was used with rosmarinic acid and both vitamins D3 and C. Antagonism was not observed in any of the cases. This data revealed the intrinsic anti-borreliae activity of doxycycline with tested phytochemicals and micronutrients indicating that their addition may enhance efficacy of this antibiotic in combating Borrelia sp. Especially the addition of flavones balcalein and luteolin to a doxycycline regimen could be explored further in defining more effective treatments against these bacteria. PMID:27570483

  18. Borrelia Spirochetes Upregulate Release and Activation of Matrix Metalloproteinase Gelatinase B (MMP-9) and Collagenase 1 (MMP-1) in Human Cells

    OpenAIRE

    Gebbia, Joseph A.; Coleman, James L.; Benach, Jorge L

    2001-01-01

    Borrelia burgdorferi, the spirochetal agent of Lyme disease, stimulated human peripheral blood monocytes to release pro-matrix metalloproteinase-9 (gelatinase B; pro-MMP-9) and active matrix metalloproteinase-1 (collagenase-1; MMP-1). Human neutrophils also released pro-MMP-9 and a 130-kDa protein with gelatinolytic activity in response to live B. burgdorferi. In addition, U937 cells and human keratinocyte cells were also stimulated to release pro-MMP-9 under the same conditions. However, hum...

  19. Evaluation of the Detection of Borrelia burgdorferi DNA in Urine Samples by Polymerase ChainReaction for Diagnosing Lyme Disease%聚合酶链法检测莱姆病患者尿液中伯氏疏螺旋体DNA(附17例报道)

    Institute of Scientific and Technical Information of China (English)

    谭毓绘; 朱沂; 刘勇; 孙荷; 龙江; 牛晓珊; 于鲁海; 莫合塔尔; 阿斯亚; 李红燕

    2012-01-01

    目的:评估聚合酶链反应(PCR)检测莱姆病患者尿液中伯氏疏螺旋体DNA的诊断价值.方法:2004至2006年来自新疆莱姆病自然疫源地临床疑似莱姆病患者17例(均有蜱暴露史和莱姆病临床证据者)和6例以前确诊并治疗后完全恢复的莱姆病患者,共23例为病例组;另选择25例非疑似莱姆病患者为对照组.应用PCR方法检测尿液中伯氏疏螺旋体DNA,采用间接免疫荧光(IFA)检测血清伯氏疏螺旋体抗体.结果:①17例莱姆病患者临床表现呈多系统损害.12/17例(70.59%)为神经型(周围神经病3例,其中1例合并下肢瘀积性皮炎;游走性红斑并发莱姆脑病2例;脑膜脑炎2例;脑炎2例;脑膜炎、脑干炎和脊髓炎各1例);流感样症状2例,其中伴皮疹1例;余心脏型、精神障碍和肝病型各1例.②PCR伯氏疏螺旋体DNA检测阳性17/17例(总阳性率100%).4例血清IFA试验伯氏疏螺旋体抗体阳性.以前确诊并治疗者6例及对照组25例尿PCR伯氏疏螺旋体DNA和血清IFA试验伯氏疏螺旋体抗体均阴性.③10例完成抗生素治疗后3个月复查,9例PCR伯氏疏螺旋体DNA检测阴性(包括3例晚期莱姆病患者),1例2年复查时仍阳性;4例血清伯氏疏螺旋体抗体阳性者复查均阴性.④早期莱姆病多采用几周的抗生素治疗方案.结论:PCR检测尿液中伯氏疏螺旋体DNA是莱姆病一个有价值的诊断工具.%Aim: To estimate the value of detecting Borrelia bugdorferi DNA in urine samples by polymerase chain reaction (PCR) for diagnosing Lyme disease. Methods: 17 patients suspected of Lyme disease come from nature foci and 25 cases with Non-Lyme disease evidence were enrolled from 2004 to 2006. Lyme serology was performed by IF A. Urine samples were examined by PCR. Results: 17 patients with Lyme disease showed sings of multisystem damage. The majority of them, accounting for 70.59%, was nervous system diseases called neuroborreliosis, which were

  20. Survey of Borreliae in ticks, canines, and white-tailed deer from Arkansas, U.S.A.

    Directory of Open Access Journals (Sweden)

    Fryxell Rebecca T

    2012-07-01

    Full Text Available Abstract Background In the Eastern and Upper Midwestern regions of North America, Ixodes scapularis (L. is the most abundant tick species encountered by humans and the primary vector of B. burgdorferi, whereas in the southeastern region Amblyomma americanum (Say is the most abundant tick species encountered by humans but cannot transmit B. burgdorferi. Surveys of Borreliae in ticks have been conducted in the southeastern United States and often these surveys identify B. lonestari as the primary Borrelia species, surveys have not included Arkansas ticks, canines, or white-tailed deer and B. lonestari is not considered pathogenic. The objective of this study was to identify Borrelia species within Arkansas by screening ticks (n = 2123, canines (n = 173, and white-tailed deer (n = 228 to determine the identity and locations of Borreliae endemic to Arkansas using PCR amplification of the flagellin (flaB gene. Methods Field collected ticks from canines and from hunter-killed white-tailed were identified to species and life stage. After which, ticks and their hosts were screened for the presence of Borrelia using PCR to amplify the flaB gene. A subset of the positive samples was confirmed with bidirectional sequencing. Results In total 53 (21.2% white-tailed deer, ten (6% canines, and 583 (27.5% Ixodid ticks (252 Ixodes scapularis, 161 A. americanum, 88 Rhipicephalus sanguineus, 50 Amblyomma maculatum, 19 Dermacentor variabilis, and 13 unidentified Amblyomma species produced a Borrelia flaB amplicon. Of the positive ticks, 324 (22.7% were collected from canines (151 A. americanum, 78 R. sanguineus, 43 I. scapularis, 26 A. maculatum, 18 D. variabilis, and 8 Amblyomma species and 259 (37.2% were collected from white-tailed deer (209 I. scapularis, 24 A. maculatum, 10 A. americanum, 10 R. sanguineus, 1 D. variabilis, and 5 Amblyomma species. None of the larvae were PCR positive. A majority of the flaB amplicons were homologous with B

  1. Transposon mutagenesis as an approach to improved understanding of Borrelia pathogenesis and biology

    Directory of Open Access Journals (Sweden)

    Tao eLin

    2014-05-01

    Full Text Available Transposon insertion provides a method for near-random mutation of bacterial genomes, and has been utilized extensively for the study of bacterial pathogenesis and biology. This approach is particularly useful for organisms that are relatively refractory to genetic manipulation, including Lyme disease Borrelia. In this review, progress to date in the application of transposon mutagenesis to the study of Borrelia burgdorferi is reported. An effective Himar1-based transposon vector has been developed and used to acquire a sequence-defined library of nearly 4,500 mutants in the infectious, moderately transformable B. burgdorferi B31 derivative 5A18NP1. Analysis of these transposon mutants using signature-tagged mutagenesis (STM and Tn-seq approaches has begun to yield valuable information regarding the genes important in the pathogenesis and biology of this organism.

  2. Seasonal distribution of Borreliae in Ixodes ricinus ticks in the Belgrade region

    Directory of Open Access Journals (Sweden)

    Milutinović Marija

    2006-01-01

    Full Text Available Green areas at four localities in the Belgrade region (Ada Ciganlija, Košutnjak, Miljakovac forest, and Mt. Avala were investigated in 2004. The aim of the research was to clarify the faunistic composition, relative abundance, and population dynamics of ticks, as well as the seasonal distribution of Borrelia burgdorferi sensu lato (sl in Ixodes ricinus. Two species of ticks were detected: Ixodes ricinus and Dermacentor reticulates. Relative abundance analysis revealed that the species Ixodes ricinus was predominant (97.41 %. Out of 942 Ixodes ricinus ticks, 188 (19.96 % were infected with Borrelia burgdorferi sl. The infection rate of adults by localities ranged from 19.16% to 30.99% (Mt. Avala and Ada Ciganlija, respectively.

  3. Laboratoriediagnostik af infektion forårsaget af Borrelia burgdorferi

    DEFF Research Database (Denmark)

    Dessau, Ram B; Bangsborg, Jette M; Jensen, Tove P Ejlertsen;

    2006-01-01

    The laboratory diagnosis of Lyme disease in Denmark is reviewed with recommendations for serological testing. In Denmark the laboratory testing is performed with an ELISA technique. Most laboratories use an assay based on purified flagella antigen. The two-tier approach with Western Blot as...... confirmatory testing is not recommended since the contribution to the diagnostic specificity is only marginal. Predictive values of Lyme serology are presented, based on the estimated prevalence of the different stages of Lyme disease in Denmark....

  4. Laboratoriediagnostik af infektion forårsaget af Borrelia burgdorferi

    DEFF Research Database (Denmark)

    Dessau, Ram; Bangsborg, Jette Marie; Jensen, Tove P Ejlertsen;

    2006-01-01

    The laboratory diagnosis of Lyme disease in Denmark is reviewed with recommendations for serological testing. In Denmark the laboratory testing is performed with an ELISA technique. Most laboratories use an assay based on purified flagella antigen. The two-tier approach with Western Blot as confi...

  5. Overexpression of CsrA (BB0184) Alters the Morphology and Antigen Profiles of Borrelia burgdorferi▿

    OpenAIRE

    Sanjuan, Eva; Maria D Esteve-Gassent; Maruskova, Mahulena; Seshu, J.

    2009-01-01

    Borrelia burgdorferi, the agent of Lyme disease, alters its gene expression in response to highly disparate environmental signals encountered in its hosts. Among the relatively few regulators of adaptive gene expression present in the borrelial genome is an open reading frame (ORF), BB0184, annotated as CsrA (carbon storage regulator A). CsrA, in several bacterial species, has been characterized as a small RNA binding protein that functions as a global regulator affecting mRNA stability or le...

  6. Conversion of a linear to a circular plasmid in the relapsing fever agent Borrelia hermsii.

    OpenAIRE

    Ferdows, M S; Serwer, P; Griess, G A; Norris, S. J.; Barbour, A G

    1996-01-01

    Spirochetes of the genus Borrelia have genomes composed of both linear and circular replicons. We characterized the genomic organization of B. burgdorferi, B. hermsii, B. turicatae, and B. anserina with pulsed-field gel electrophoresis. All four species contained a linear chromosome approximately 1 Mb in size and multiple linear plasmids in the 16- to 200-kb size range. Plasmids 180 and 170 kb in size, present in the relapsing fever agents B. hermsii and B. turicatae but not in the other two ...

  7. Longitudinal analysis of tick densities and Borrelia, Anaplasma, and Ehrlichia infections of Ixodes ricinus ticks in different habitat areas in The Netherlands.

    NARCIS (Netherlands)

    Wielinga, Peter R; Gaasenbeek, Cor; Fonville, Manoj; Boer, Albert de; Vries, Ankje de; Dimmers, Wim; Akkerhuis Op Jagers, Gerard; Schouls, Leo M; Borgsteede, Fred; Giessen, Joke W B van der

    2006-01-01

    From 2000 to 2004, ticks were collected by dragging a blanket in four habitat areas in The Netherlands: dunes, heather, forest, and a city park. Tick densities were calculated, and infection with Borrelia burgdorferi and Anaplasma and Ehrlichia species was investigated by reverse line blot analysis.

  8. Longitudinal analysis of tick densities and Borrelia, Anaplasma, and Ehrlichia infections of Ixodes ricinus ticks in different habitat areas in the Netherlands

    NARCIS (Netherlands)

    Wielinga, P.R.; Gaasenbeek, C.P.H.; Fonville, M.; Boer, de A.G.; Vries, de A.; Dimmers, W.J.; Jagers op Akkerhuis, G.A.J.M.; Schouls, L.M.; Borgsteede, F.H.M.; Giessen, van der J.W.B.

    2006-01-01

    From 2000 to 2004, ticks were collected by dragging a blanket in four habitat areas in The Netherlands: dunes, heather, forest, and a city park. Tick densities were calculated, and infection with Borrelia burgdorferi and Anaplasma and Ehrlichia species was investigated by reverse line blot analysis.

  9. Borrelia bavariensis sp. nov. is widely distributed in Europe and Asia.

    Science.gov (United States)

    Margos, Gabriele; Wilske, Bettina; Sing, Andreas; Hizo-Teufel, Cecilia; Cao, Wu-Chun; Chu, Chenyi; Scholz, Holger; Straubinger, Reinhard K; Fingerle, Volker

    2013-11-01

    Since the original description of Borrelia bavariensis sp. nov. in 2009, additional samples available from humans and ticks from Europe and Mongolia, respectively, have been used to further characterize Borrelia strains belonging to this group of spirochaetes that utilize rodents as reservoir hosts. These investigations suggested the presence of related strains in Europe and Asia and confirmed their status as representing a distinct species. Furthermore, samples that were investigated by researchers from China and Japan confirm the ecological relationship of members of this proposed species with rodents and suggest that it has a wide distribution in Eurasia. Here, we use phylogenetic and genetic distance analyses to validate B. bavariensis sp. nov. as a species within the Borrelia burgdorferi sensu lato species complex. The type strain is PBi(T) ( = DSM 23469(T) = BAA-2496(T)). PMID:23838444

  10. Natural infection of 4-Lyme disease Borrelia burgdorferi in Diebu and Huajian areas of Gansu province%甘肃省迭部和铧尖地区4种蜱自然感染莱姆病伯氏疏螺旋体的调查

    Institute of Scientific and Technical Information of China (English)

    罗晓红; 何宗平

    2011-01-01

    目的 调查甘肃省迭部和铧尖地区4种蜱自然感染莱姆病伯氏疏螺旋体(Bb)情况,为莱姆病的防治提供科学依据。方法 2010年3月至6月,在甘肃省迭部和铧尖地区岷山北麓迭部林区(秦岭山脉)和肃南祁连山北麓铧尖林区,对4种优势蜱(森林革蜱、草原革蜱、日本血蜱和青海血蜱)自然感染莱姆病Bb进行流行病学检测。采用夹夜法,每隔10 m布夹,晚放晨收,将捕获的啮齿类动物逆毛检虫法采集寄生蜱,同时采用拖旗法采集游离蜱。对采集的4种活的成蜱,清洗消毒后解剖取其中肠内容物分别涂片,置暗视野镜下观察莱姆病Bb;对所分离到的螺旋体再用Bb单克隆和多克隆抗体鉴别试验加以证实。结果 共采集到蜱类2科8属36种,即硬蜱科6属33种,软蜱科2属3种。解剖森林革蜱、草原革蜱、日本血蜱和草原硬蜱4种201只蜱的肠道,暗视野观察出携带莱姆病Bb的阳性蜱25只,阳性率为12.44%(25/201);接种培养森林革蜱、草原革蜱和日本血蜱3种65只蜱,从12只蜱体内培养分离出莱姆病Bb,阳性率为18.46%(12/65)。结论 森林革蜱、草原革蜱和日本血蜱均有程度不同地莱姆病Bb的自然感染。%Objective In order to offer scientific evidence for prevention and therapy of Lyme disease, we had investigated the natural infection of lyme disease Borrelia burgdorferi(Bb) of 4 ticks in Diebu and Huajian areas of Gansu province. Methods Epidemiology detection of natural infection of lyme disease Bb was accomplished for four dominant tick ( Dermacetor silvarum , Dermacentor nuttalli , Haemaphysalis Japonica and Ixodes Crenulatus ) at Diebu, Diebu forest zone(Qin mountain), north of Min Mountains of Huajian region and Huajian forest zone, north of Qilian Mountains of Sunan region, Gansu, from March to June, 2010. The methods of dispersion of clamp every 10 m which was put in the morning and retrieved at

  11. Isolation and characterization of channel-forming proteins in the outer membrane of E. coli and Borrelia species

    OpenAIRE

    Denker, Katrin

    2006-01-01

    In this study pore forming proteins of the gram-negative bacteria B. burgdorferi, B. duttonii and E.coli were investigated. Therefore the study is subdivided into three parts. In the first part outer membrane preparation of three relapsing fever Borrelia were investigated. In the second part the putative TolC homologue BB0124 of B. burgdorferi, the Lyme borreliosis agent, was studied. In the last part the influence of point mutants within the greasy slide of the maltose specific porin (LamB) ...

  12. Investigation ofBorrelia spp. in ticks (Acari:Ixodidae) at the border crossings between China and Russia in Heilongjiang Province, China

    Institute of Scientific and Technical Information of China (English)

    Shi Liu; Chao Yuan; Yun-Fu Cui; Bai-Xiang Li; Li-Jie Wu; Ying Liu

    2012-01-01

    Objective:To investigate the precise species of tick vector and theBorrelia spirochete pathogen at the Heilongjiang Province international border with Russia.Methods:In this study, ticks were collected from12 Heilongjiang border crossings (including grasslands, shrublands, forests, and plantantions) to determine the rate and species type of spirochete-infected ticks and the most prevalent spirochete genotypes.Results:The ticks represented three genera and four species of the Ixodidae family [Ixodes persulcatus,Dermacentor silvarum,Haemaphysalis concinna and Haemaphysalis japonica].Ixodes persulcatus had the highest amount ofBorrelia burgdorferi sensu lato infection of25.6% and the most common species ofBorreliaisolated from Ixodes persulcatus wasBorrelia garinii, strainPD91.Conclusions: Our results suggest thatBorrelia gariniiPD91-infectedIxodes persulcatus may be the principal cause of Lyme disease in the border crossing areas of Heilongjiang Province.

  13. Borreliacidal activity of Borrelia metal transporter A (BmtA binding small molecules by manganese transport inhibition

    Directory of Open Access Journals (Sweden)

    Wagh D

    2015-02-01

    Full Text Available Dhananjay Wagh,* Venkata Raveendra Pothineni,* Mohammed Inayathullah, Song Liu, Kwang-Min Kim, Jayakumar Rajadas Biomaterials and Advanced Drug Delivery Laboratory, Stanford Cardiovascular Pharmacology Division, Cardiovascular Institute, Stanford University School of Medicine, Palo Alto, CA, USA *These authors contributed equally to this work  Abstract: Borrelia burgdorferi, the causative agent of Lyme disease, utilizes manganese (Mn for its various metabolic needs. We hypothesized that blocking Mn transporter could be a possible approach to inhibit metabolic activity of this pathogen and eliminate the infection. We used a combination of in silico protein structure prediction together with molecular docking to target the Borrelia metal transporter A (BmtA, a single known Mn transporter in Borrelia and screened libraries of FDA approved compounds that could potentially bind to the predicted BmtA structure with high affinity. Tricyclic antihistamines such as loratadine, desloratadine, and 3-hydroxydesloratadine as well as yohimbine and tadalafil demonstrated a tight binding to the in silico folded BmtA transporter. We, then, tested borreliacidal activity and dose response of the shortlisted compounds from this screen using a series of in vitro assays. Amongst the probed compounds, desloratadine exhibited potent borreliacidal activity in vitro at and above 78 µg/mL (250 µM. Borrelia treated with lethal doses of desloratadine exhibited a significant loss of intracellular Mn specifically and a severe structural damage to the bacterial cell wall. Our results support the possibility of developing a novel, targeted therapy to treat Lyme disease by targeting specific metabolic needs of Borrelia.  Keywords: Lyme disease, BmtA, Borrelia burgdorferi, desloratadine, Bac Titer-Glo assay

  14. 我国滇西北山区家犬莱姆病螺旋体基因的检测及序列分析%Detection and sequence analysis of Borrelia burgdorferi sensu lato in dogs from north-western Yunnan province,China

    Institute of Scientific and Technical Information of China (English)

    边长玲; 龚正达; 张丽云; 李四全; 李璋鸿; 褚辰一; 乔文清; 和志炎

    2010-01-01

    目的 了解滇西北山区莱姆病伯氏疏螺旋体的自然感染及基因分型情况.方法 以山区农村家犬作为指示动物,使用真空采血管抽取家犬全血,应用巢式PCR扩增犬血中伯氏疏螺旋体5S~23S rRNA间隔区片段,然后对阳性片段进行测序,并将所测序列与GenBank中注册的基因序列进行比较分析.结果共检测贡山、福贡2个县3个乡镇山村63只家犬血液,其中发现9只犬为阳性,阳性率14.29%.基因序列分析结果显示,当地犬感染的伯氏疏螺旋体为Borrelia garinii.结论 首次证实云南省西北部山区家犬中存在莱姆病Borrelia garinii型伯氏疏螺旋体感染.

  15. 中国莱姆病螺旋体PD91外膜蛋白A的克隆表达及免疫保护性的初步研究%Molecular cloning and expression of OspA of a Chinese Borrelia burgdorferi PD91 and preliminary research on the protectivity of rOspA

    Institute of Scientific and Technical Information of China (English)

    郝琴; 万康林

    2002-01-01

    目的克隆并表达中国莱姆病螺旋体Borrelia garinii基因型代表菌株PD91的外膜蛋白A(OspA),并对其免疫保护性进行初步研究,为进一步研制莱姆病疫苗提供基础. 方法用聚合酶链反应(PCR)从莱姆病螺旋体PD91全基因组DNA中将OspA基因调出,插入原核表达载体P42,在大肠杆菌BL21(DE3)中表达,表达产物用SDS-PAGE、Western blot分析,并进行基因序列测定.用重组OspA(rOspA)免疫新西兰家兔,用间接免疫荧光(IFA)检测其血清特异性抗体(IgG),并进行体外中和试验,从而对其免疫保护性有初步的了解. 结果 rOspA在宿主菌内表达高效、稳定;Western blot显示其与抗OspA的多抗有较好的免疫反应性;用rOspA免疫新西兰家兔后,其血清抗体(IgG)效价显著升高(32倍),体外中和试验表明,每毫升兔抗rOspA血清可杀灭105个莱姆病螺旋体. 结论在国内首次成功地对中国莱姆病螺旋体Borrelia garinii基因型的OspA基因进行了克隆和表达.rOspA有较好的免疫保护性,可作为多价莱姆病疫苗的一种成分.

  16. Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7

    Energy Technology Data Exchange (ETDEWEB)

    Dunn, J.J.; Barbour, A.G.

    1996-11-05

    A method is provided for preparing soluble recombinant variations of Borrelia lipoproteins such as Borrelia burgdorferi outer surface protein A (OspA) and outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The method includes synthesizing a set of oligonucleotide primers, amplifying the template DNA utilizing the PCR, purifying the amplification products, cloning the amplification products into a suitable expression vector, transforming a suitable host utilizing the cloned expression vector, cultivating the transformed host for protein production and subsequently isolating and purifying the resulting protein. Also provided are soluble, recombinant variations of Borrelia burgdorferi outer surface protein A (OspA), outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The expression vectors harboring DNA encoding the recombinant variations, pET9-OspA, pET9-OspB and pET9-Vmp7, as well as the E. coli host BL21(DE3)/pLysS transformed with each of these vectors, are also disclosed. 38 figs.

  17. Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7

    Energy Technology Data Exchange (ETDEWEB)

    Dunn, John J. (Bellport, NY); Barbour, Alan G. (San Antonio, TX)

    1996-11-05

    A method is provided herein for preparing soluble recombinant variations of Borrelia lipoproteins such as Borrelia burgdorferi outer surface protein A (OspA) and outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The method includes synthesizing a set of oligonucleotide primers, amplifying the template DNA utilizing the PCR, purifying the amplification products, cloning the amplification products into a suitable expression vector, transforming a suitable host utilizing the cloned expression vector, cultivating the transformed host for protein production and subsequently isolating and purifying the resulting protein. Also provided are soluble, recombinant variations of Borrelia burgdorferi outer surface protein A (OspA), outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The expression vectors harboring DNA encoding the recombinant variations, pET9-OspA, pET9-OspB and pET9-Vmp7, as well as the E. coli host BL21(DE3)/pLysS transformed with each of these vectors, are also disclosed.

  18. Detection of Borreliae in Archived Sera from Patients with Clinically Suspect Lyme Disease

    Directory of Open Access Journals (Sweden)

    Sin Hang Lee

    2014-03-01

    Full Text Available The diagnoses of Lyme disease based on clinical manifestations, serological findings and detection of infectious agents often contradict each other. We tested 52 blind-coded serum samples, including 20 pre-treatment and 12 post-treatment sera from clinically suspect Lyme disease patients, for the presence of residual Lyme disease infectious agents, using nested PCR amplification of a signature segment of the borrelial 16S ribosomal RNA gene for detection and direct DNA sequencing of the PCR amplicon for molecular validation. These archived sera were split from the samples drawn for the 2-tier serology tests performed by a CDC-approved laboratory, and are used as reference materials for evaluating new diagnostic reagents. Of the 12 post-treatment serum samples, we found DNA evidence of a novel borrelia of uncertain significance in one, which was also positive for the 2-tier serology test. The rest of the post-treatment sera and all 20 control sera were PCR-negative. Of the 20 pre-treatment sera from clinically suspect early Lyme disease patients, we found Borrelia miyamotoi in one which was 2-tier serology-negative, and a Borrelia burgdorferi in two—one negative and one positive for 2-tier serology. We conclude that a sensitive and reliable DNA-based test is needed to support the diagnosis of Lyme disease and Lyme disease-like borreliosis.

  19. Growth, cysts and kinetics of Borrelia garinii (Spirochaetales: Spirochaetacea in different culture media

    Directory of Open Access Journals (Sweden)

    Angela de Oliveira

    2010-08-01

    Full Text Available The aim of the present paper was to evaluate cyst formation and growth parameters of Borrelia garinii in a range of media differing in formulation and cost. A qualitative assessment of morphology and motility of B. garinii was conducted. All media were prepared aseptically and used in test tubes or Petri dishes. For each medium, the initial spirochete concentration was standardized to 10³ spirochets/mL. The following culture media were suitable to grow B. garinii: Barbour-Stoenner-Kelly, brain heart infusion and PMR. Growth was minimal at six weeks post-inoculation and maximum spirochete density was observed between 9-12 weeks. Often, the cultures developed cysts of different sizes, isolated or in groups, with a spiraled portion of variable sizes, mainly in unfavorable culture media. Brazilian Lyme disease-like illness, also known as Baggio-Yoshinari syndrome (BYS, is a new and interesting emerging tick-borne disease, caused by Borrelia burgdorferi sensu lato spirochetes, only during its cystic forms. It has been assumed that the peculiar clinical and laboratory features of BYS are consequential to the absence of a human sucker Ixodes ricinus complex tick at risk areas in Brazil, supporting the concept that the borrelia phenotypic expression pattern is modified as it is transmitted through the host.

  20. Borrelia hispanica relapsing fever, Morocco.

    Science.gov (United States)

    Sarih, M'hammed; Garnier, Martine; Boudebouch, Najma; Bouattour, Ali; Rihani, Abdelaziz; Hassar, Mohammed; Gern, Lise; Postic, Danièle; Cornet, Muriel

    2009-10-01

    We found that 20.5% of patients with an unexplained fever in northwestern Morocco had tick-borne relapsing fever. Molecular detection specific for the 16S rRNA gene identified Borrelia hispanica. The noncoding intergenic spacer sequence domain showed high sensitivity and good resolution for this species. PMID:19861058

  1. Borrelia hispanica Relapsing Fever, Morocco

    OpenAIRE

    Sarih, M’hammed; Garnier, Martine; Boudebouch, Najma; Bouattour, Ali; Rihani, Abdelaziz; Hassar, Mohammed; Gern, Lise; Postic, Danièle; Cornet, Muriel

    2009-01-01

    We found that 20.5% of patients with an unexplained fever in northwestern Morocco had tick-borne relapsing fever. Molecular detection specific for the 16S rRNA gene identified Borrelia hispanica. The noncoding intergenic spacer sequence domain showed high sensitivity and good resolution for this species.

  2. Beta-Amyloid Deposition and Alzheimer's Type Changes Induced by Borrelia Spirochetes

    Energy Technology Data Exchange (ETDEWEB)

    Miklossy,J.; Kis, A.; Radenovic, A.; Miller, L.; Forro, L.; Martins, R.; Reiss, K.; Darbinian, N.; Darekar, P.; et al.

    2006-01-01

    The pathological hallmarks of Alzheimer's disease (AD) consist of {beta}-amyloid plaques and neurofibrillary tangles in affected brain areas. The processes, which drive this host reaction are unknown. To determine whether an analogous host reaction to that occurring in AD could be induced by infectious agents, we exposed mammalian glial and neuronal cells in vitro to Borrelia burgdorferi spirochetes and to the inflammatory bacterial lipopolysaccharide (LPS). Morphological changes analogous to the amyloid deposits of AD brain were observed following 2-8 weeks of exposure to the spirochetes. Increased levels of {beta}-amyloid presursor protein (A{beta}PP) and hyperphosphorylated tau were also detected by Western blots of extracts of cultured cells that had been treated with spirochetes or LPS. These observations indicate that, by exposure to bacteria or to their toxic products, host responses similar in nature to those observed in AD may be induced.

  3. Complement factor H binding by different Lyme disease and relapsing fever Borrelia in animals and human

    Directory of Open Access Journals (Sweden)

    Gil Horacio

    2009-07-01

    Full Text Available Abstract Background Borreliae employ multiple immune evasive strategies such as binding to complement regulatory proteins [factor H (fH and factor H like-1 (FHL1], differential regulation of surface membrane proteins, antigenic variation, and binding of plasminogen/plasmin and matrix metalloproteinases. As a complement regulatory subunit, fH serves as a cofactor for the factor I-mediated cleavage of C3b. fH binding by Borrelia has been correlated with pathogenesis as well as with host diversity. Here we show the differential binding of borrelial proteins to fH from human and animal sera. Findings Affinity ligand binding experiments, 2-D electrophoresis, and protein identification and peptide de novo sequencing based on mass spectrometry, revealed novel fH putative binding proteins of Lyme- and relapsing fever Borrelia. An OspA serotype-associated differential human and animal fH binding by B. garinii was also observed, which could be related with the ability of some strains from serotypes 4 and 7 to invade non-nervous system tissues. Also, the variable affinity of binding proteins expressed by different Borrelia to animal fH correlated with their host selectivity. Conclusion The novel animal and human putative fH binding proteins (FHBPs in this study underscore the importance of evasion of complement in the pathogenesis of Borrelia infections.

  4. Characterization of a DNA Adenine Methyltransferase Gene of Borrelia hermsii and Its Dispensability for Murine Infection and Persistence

    Science.gov (United States)

    James, Allison E.; Rogovskyy, Artem S.; Crowley, Michael A.; Bankhead, Troy

    2016-01-01

    DNA methyltransferases have been implicated in the regulation of virulence genes in a number of pathogens. Relapsing fever Borrelia species harbor a conserved, putative DNA methyltransferase gene on their chromosome, while no such ortholog can be found in the annotated genome of the Lyme disease agent, Borrelia burgdorferi. In the relapsing fever species Borrelia hermsii, the locus bh0463A encodes this putative DNA adenine methyltransferase (dam). To verify the function of the BH0463A protein product as a Dam, the gene was cloned into a Dam-deficient strain of Escherichia coli. Restriction fragment analysis subsequently demonstrated that complementation of this E. coli mutant with bh0463A restored adenine methylation, verifying bh0463A as a Dam. The requirement of bh0463A for B. hermsii viability, infectivity, and persistence was then investigated by genetically disrupting the gene. The dam- mutant was capable of infecting immunocompetent mice, and the mean level of spirochetemia in immunocompetent mice was not significantly different from wild type B. hermsii. Collectively, the data indicate that dam is dispensable for B. hermsii viability, infectivity, and persistence. PMID:27195796

  5. Detection of Borrelia, Ehrlichia, and Rickettsia spp. in ticks in northeast Missouri.

    Science.gov (United States)

    Hudman, D A; Sargentini, N J

    2016-07-01

    We evaluated Amblyomma americanum (lone star tick) and Dermacentor variabilis (American dog tick) in northeast Missouri for the presence of Borrelia, Ehrlichia, and Rickettsia bacteria. We collected actively questing ticks from four sites within Adair County, Missouri. A total of 15,162 ticks were collected, of which 13,980 were grouped in 308 pools (lone star ticks, 288 pools; American dog ticks, 20 pools) and tested for presence/absence of bacteria using polymerase chain reaction. Infection rates were calculated as the maximum likelihood estimation (MLE) with 95% confidence intervals (CI). Of the 308 pools tested, 229 (74.4%) were infected with bacteria and the overall MLE of the infection rate per 100 ticks was calculated as 2.9% (CI 2.61-3.21). Infection rates varied among life stages, 28.6% (CI 23.89-33.97) in adults, 7.0% (CI 5.10-9.86) in nymphs, and 1.0% (CI 0.75-1.20) in larvae. In the 116 adult lone star pools, infection rates were calculated for Borrelia lonestari (1.4%), Borrelia spp. (2.7%), Ehrlichia chaffeensis (6.1%), Ehrlichia ewingii (3.3%), Rickettsia amblyommii (18.3%), and Rickettsia montanensis (0.4%). Infection rates for the 52 nymphal lone star pools were calculated as B. lonestari (1.03%), Borrelia spp. (0.40%), E. chaffeensis (2.02%), E. ewingii (0.24%), and R. amblyommii (2.70%). In the 20 adult American dog tick pools, infection rates were determined as E. chaffeensis (9.47%), E. ewingii (5.47%), and R. montanensis (8.06%). Eight Borrelia samples were sequenced with five 99-100% identical to B. burgdorferi (s.l.) and three 99% identical to B. lonestari. Eight samples were sequenced for E. chaffeensis (all 99-100% identical) and one sample was sequenced for E. ewingii (99% identical). Seven samples were sequenced for Rickettsia and three were 99% identical to R. montanensis and four were 100% identical to R. amblyommii. This study demonstrates B. lonestari, E. chaffeensis, E. ewingii, R. amblyommii, and R. montanensis in northeast

  6. Induktion von Matrixmetalloproteinasen und Interleukin-18 durch Borrelia burgdorferi in vivo und in vitro

    OpenAIRE

    Unsicker, Catrin

    2010-01-01

    Lyme borreliosis is a systemic disease involving the skin, the musculoskeletal system, the heart and the central nervous system. The shift of the cytokine pattern towards the Th1 response is one characteristic feature of the disease. Recently, interleukin-18 has been described to induce interferon-gamma. Therefore the potential role of IL-18 in Lyme disease has been studied. In addition, it is not clear how the spirochetes can disseminate within the host organism...

  7. Population Bottlenecks during the Infectious Cycle of the Lyme Disease Spirochete Borrelia burgdorferi

    Czech Academy of Sciences Publication Activity Database

    Rego, Ryan O. M.; Bestor, A.; Štefka, Jan; Rosa, P. A.

    2014-01-01

    Roč. 9, č. 6 (2014), e101009. E-ISSN 1932-6203 Institutional support: RVO:60077344 Keywords : sensu stricto * mammalian host * peromyscus-leucopus * Ixodes ricinus * ticks * mice * transmission * dissemination * diversity * North America Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 3.234, year: 2014

  8. 伯氏螺旋体在小鼠宿主中的关节特异性转录谱%The Joint-specific Expression Profile of Borrelia burgdorfri in the Murine Hosts

    Institute of Scientific and Technical Information of China (English)

    宝福凯; Erol Fikerig

    2008-01-01

    目的 建立伯氏疏螺旋体感染的小鼠模型,研究伯氏疏螺旋体在小鼠关节组织中的关节特异性基因表达谱.方法 首先,建立伯氏疏螺旋体感染小鼠模型,在不同时间点处死小鼠,收集小鼠关节、心脏、皮肤和膀胱组织,从四种组织中分别提取总RNA;随后,用DECAL技术和微阵列技术分析不同时间点伯氏疏螺旋体在四种组织中的转录组;最后,将伯氏疏螺旋体在关节中的转录组依次与其他三种组织中的转录组进行比较,找出仅在关节中表达的伯氏疏螺旋体基因,从而获得伯氏疏螺旋体不同时间点的关节特异性基因表达谱.结果 与其他组织相比,在感染后第15天,伯氏疏螺旋体在小鼠关节组织特异地表达21个基因,其中13个基因位于伯氏疏螺旋体染色体上,8个基因位于质粒上;在感染后的第105天,伯氏疏螺旋体在小鼠关节组织特异地表达24个基因,其中13个基因位于伯氏疏螺旋体染色体上,11个基因位于质粒上.结论 伯氏疏螺旋体在小鼠关节组织中存在独特的基因表达谱,这些在关节中特异表达的基因可能与莱姆关节炎的发生、发展有关.%Objective To identify the joint-specific expression profile of Borrelia burgdorfri in the routine hosts.Methods Establishing the routine model of Lyme disease,sacrificing the infected mice at different time-points,collecting the joint,heart,skin,and urinary bladder samples of the infected mice,and extracting total RNA from the samples.DECAL technique and microarray were applied to btain the transcriptomes of Borrelia burgdorferi in the joints,heart,skin,and urinary bladder,the transcriptomes from different tissues were compared to identify the joint-specific expression profile of Borrelia burgdorferi.Results Borrelia burgdorferi expresses 21 joint-specific genes,including 13 genes located in chromosome and 9 genes located in plasmids at day 15 after infection,and expresses 24 genes

  9. Transstadial Transmission of Borrelia turcica in Hyalomma aegyptium Ticks

    OpenAIRE

    Zsuzsa Kalmár; Vasile Cozma; Hein Sprong; Setareh Jahfari; Gianluca D'Amico; Mărcuțan, Daniel I.; Ionică, Angela M.; Cristian Magdaş; David Modrý; Mihalca, Andrei D

    2015-01-01

    Borrelia turcica comprises the third major group of arthropod-transmitted borreliae and is phylogenetically divergent from other Borrelia groups. The novel group of Borrelia was initially isolated from Hyalomma aegyptium ticks in Turkey and it was recently found in blood and multiple organs of tortoises exported from Jordan to Japan. However, the ecology of these spirochetes and their development in ticks or the vertebrate hosts were not investigated in detail; our aims were to isolate the pa...

  10. DETECTION OF BORRELIA BURGDOFERI DNA IN GRANULOMATOUS TISSUES FROM PATIENTSWITH SARCOIDOSIS USING POLYMERASE CHAIN REACTION IN SITU TECHNIQUE

    Institute of Scientific and Technical Information of China (English)

    徐作军; 马东来; 罗慰慈; 朱元珏

    1996-01-01

    To investigate the correlation between sarcoidosis and Borrelia burgdorferi (Bb) infection,flagella DNA of Bb were detected in 23 granulomatous tissue specimens from patients with confirmed sarcoidosis usingpolymerase chain reaction in situ technique (in situ PCR) and the antibodies to Bb were examined in 55 serum samples obtained from the patients by indirect immunoflurescence assays. Our data presented that =(1) None of granulomatous tissues was found to have Bb DNA in 23 tissue samples. (2) Thirty of 55(54.6%) patients with sarcoidosis were found antibodies to Bh positive,in contrast,six of 60 (10%) norreal subjects had antibodies against Bb,the positive rate was remarkably higher in patient group than thatin healthy group (P(0. 005). The results suggest that Bb might not be the causative agent of sarcoidosis,the elevated titres of serum antibodies against Bb in patients with sarcoidosis is a nonspecific response.

  11. Complement factor H binding by different Lyme disease and relapsing fever Borrelia in animals and human

    OpenAIRE

    Gil Horacio; Camafeita Emilio; Escudero Raquel; Bhide Mangesh R; Jado Isabel; Anda Pedro

    2009-01-01

    Abstract Background Borreliae employ multiple immune evasive strategies such as binding to complement regulatory proteins [factor H (fH) and factor H like-1 (FHL1)], differential regulation of surface membrane proteins, antigenic variation, and binding of plasminogen/plasmin and matrix metalloproteinases. As a complement regulatory subunit, fH serves as a cofactor for the factor I-mediated cleavage of C3b. fH binding by Borrelia has been correlated with pathogenesis as well as with host diver...

  12. Lyme disease and relapsing fever Borrelia elongate through zones of peptidoglycan synthesis that mark division sites of daughter cells.

    Science.gov (United States)

    Jutras, Brandon Lyon; Scott, Molly; Parry, Bradley; Biboy, Jacob; Gray, Joe; Vollmer, Waldemar; Jacobs-Wagner, Christine

    2016-08-16

    Agents that cause Lyme disease, relapsing fever, leptospirosis, and syphilis belong to the phylum Spirochaetae-a unique lineage of bacteria most known for their long, spiral morphology. Despite the relevance to human health, little is known about the most fundamental aspects of spirochete growth. Here, using quantitative microscopy to track peptidoglycan cell-wall synthesis, we found that the Lyme disease spirochete Borrelia burgdorferi displays a complex pattern of growth. B. burgdorferi elongates from discrete zones that are both spatially and temporally regulated. In addition, some peptidoglycan incorporation occurs along the cell body, with the notable exception of a large region at the poles. Newborn cells inherit a highly active zone of peptidoglycan synthesis at midcell that contributes to elongation for most of the cell cycle. Concomitant with the initiation of nucleoid separation and cell constriction, second and third zones of elongation are established at the 1/4 and 3/4 cellular positions, marking future sites of division for the subsequent generation. Positioning of elongation zones along the cell is robust to cell length variations and is relatively precise over long distances (>30 µm), suggesting that cells ‟sense" relative, as opposed to absolute, cell length to establish zones of peptidoglycan synthesis. The transition from one to three zones of peptidoglycan growth during the cell cycle is also observed in relapsing fever Borrelia. However, this mode of growth does not extend to representative species from other spirochetal genera, suggesting that this distinctive growth mode represents an evolutionary divide in the spirochete phylum. PMID:27506799

  13. Search for Borrelia sp. in ticks collected from potential reservoirs in an urban forest reserve in the State of Mato Grosso do Sul, Brazil: a short report

    Directory of Open Access Journals (Sweden)

    IP da Costa

    2002-07-01

    Full Text Available A total of 128 ticks of the genus Amblyomma were recovered from 5 marsupials (Didelphis albiventris - with 4 recaptures - and 17 rodents (16 Bolomys lasiurus and 1 Rattus norvegicus captured in an urban forest reserve in Campo Grande, State of Mato Grosso do Sul, Brazil. Of the ticks collected, 95 (78.9% were in larval form and 22 (21.1% were nymphs; the only adult (0.8% was identified as A. cajennense. Viewed under dark-field microscopy in the fourth month after seeding, 9 cultures prepared from spleens and livers of the rodents, blood of the marsupials, and macerates of Amblyomma sp. nymphs revealed spiral-shaped, spirochete-like structures resembling those of Borrelia sp. Some of them showed little motility, while others were non-motile. No such structures could be found either in positive Giemsa-stained culture smears or under electron microscopy. No PCR amplification of DNA from those cultures could be obtained by employing Leptospira sp., B. burgdorferi, and Borrelia sp. primers. These aspects suggest that the spirochete-like structures found in this study do not fit into the genera Borrelia or Leptospira, requiring instead to be isolated for proper identification.

  14. High seroprevalence of Borrelia miyamotoi antibodies in forestry workers and individuals suspected of human granulocytic anaplasmosis in the Netherlands.

    Science.gov (United States)

    Jahfari, S; Herremans, T; Platonov, A E; Kuiper, H; Karan, L S; Vasilieva, O; Koopmans, M P G; Hovius, J W R; Sprong, H

    2014-09-01

    Substantial exposure to Borrelia miyamotoi occurs through bites from Ixodes ricinus ticks in the Netherlands, which also transmit Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. Direct evidence for B. miyamotoi infection in European populations is scarce. A flu-like illness with high fever, resembling human granulocytic anaplasmosis, has been attributed to B. miyamotoi infections in relatively small groups. Borrelia miyamotoi infections associated with chronic meningoencephalitis have also been described in case reports. Assuming that an IgG antibody response against B. miyamotoi antigens reflects (endured) infection, the seroprevalence in different risk groups was examined. Sera from nine out of ten confirmed B. miyamotoi infections from Russia were found to be positive with the recombinant antigen used, and no significant cross-reactivity was observed in secondary syphilis patients. The seroprevalence in blood donors was set at 2.0% (95% CI 0.4-5.7%). Elevated seroprevalences in individuals with serologically confirmed, 7.4% (2.0-17.9%), or unconfirmed, 8.6% (1.8-23%), Lyme neuroborreliosis were not significantly different from those in blood donors. The prevalence of anti-B. miyamotoi antibodies among forestry workers was 10% (5.3-16.8%) and in patients with serologically unconfirmed but suspected human granulocytic anaplasmosis was 14.6% (9.0-21.8%); these were significantly higher compared with the seroprevalence in blood donors. Our findings indicate that infections with B. miyamotoi occur in tick-exposed individuals in the Netherlands. In addition, B. miyamotoi infections should be considered in patients reporting tick bites and febrile illness with unresolved aetiology in the Netherlands, and other countries where I. ricinus ticks are endemic. PMID:25356364

  15. High seroprevalence of Borrelia miyamotoi antibodies in forestry workers and individuals suspected of human granulocytic anaplasmosis in the Netherlands

    Directory of Open Access Journals (Sweden)

    S. Jahfari

    2014-09-01

    Full Text Available Substantial exposure to Borrelia miyamotoi occurs through bites from Ixodes ricinus ticks in the Netherlands, which also transmit Borrelia burgdorferi sensu lato and Anaplasma phagocytophilum. Direct evidence for B. miyamotoi infection in European populations is scarce. A flu-like illness with high fever, resembling human granulocytic anaplasmosis, has been attributed to B. miyamotoi infections in relatively small groups. Borrelia miyamotoi infections associated with chronic meningoencephalitis have also been described in case reports. Assuming that an IgG antibody response against B. miyamotoi antigens reflects (endured infection, the seroprevalence in different risk groups was examined. Sera from nine out of ten confirmed B. miyamotoi infections from Russia were found to be positive with the recombinant antigen used, and no significant cross-reactivity was observed in secondary syphilis patients. The seroprevalence in blood donors was set at 2.0% (95% CI 0.4–5.7%. Elevated seroprevalences in individuals with serologically confirmed, 7.4% (2.0–17.9%, or unconfirmed, 8.6% (1.8–23%, Lyme neuroborreliosis were not significantly different from those in blood donors. The prevalence of anti-B. miyamotoi antibodies among forestry workers was 10% (5.3–16.8% and in patients with serologically unconfirmed but suspected human granulocytic anaplasmosis was 14.6% (9.0–21.8%; these were significantly higher compared with the seroprevalence in blood donors. Our findings indicate that infections with B. miyamotoi occur in tick-exposed individuals in the Netherlands. In addition, B. miyamotoi infections should be considered in patients reporting tick bites and febrile illness with unresolved aetiology in the Netherlands, and other countries where I. ricinus ticks are endemic.

  16. Assessing the abundance, seasonal questing activity, and Borrelia and tick-borne encephalitis virus (TBEV) prevalence of Ixodes ricinus ticks in a Lyme borreliosis endemic area in Southwest Finland.

    Science.gov (United States)

    Sormunen, Jani J; Klemola, Tero; Vesterinen, Eero J; Vuorinen, Ilppo; Hytönen, Jukka; Hänninen, Jari; Ruohomäki, Kai; Sääksjärvi, Ilari E; Tonteri, Elina; Penttinen, Ritva

    2016-02-01

    Studies have revealed that Ixodes ricinus (Acari: Ixodidae) have become more abundant and their geographical distribution extended northwards in some Nordic countries during the past few decades. However, ecological data of tick populations in Finland are sparse. In the current study, I. ricinus abundance, seasonal questing activity, and their Borrelia spp. and tick-borne encephalitis virus (TBEV) prevalence were evaluated in a Lyme borreliosis endemic area in Southwest Finland, Seili Island, where a previous study mapping tick densities was conducted 12 years earlier. A total of 1940 ticks were collected from five different biotopes by cloth dragging during May-September 2012. The overall tick density observed was 5.2 ticks/100m(2) for nymphs and adults. Seasonal questing activity of ticks differed between biotopes and life stages: bimodal occurrences were observed especially for nymphal and adult ticks in forested biotopes, while larvae in pastures exhibited mostly unimodal occurrence. Prevalence of Borrelia and TBEV in ticks was evaluated using conventional and real-time PCR. All samples were negative for TBEV. Borrelia prevalence was 25.0% for adults (n=44) and the minimum infection rate (MIR) 5.6% for pooled nymph samples (191 samples, 1-14 individuals per sample; 30/191 positive). No Borrelia were detected in pooled larval samples (63 samples, 1-139 individuals per sample). Five species of Borrelia were identified from the samples: B. afzelii, B. burgdorferi s.s., B. garinii, B. valaisiana and B. miyamotoi. In Finland, B. valaisiana and B. miyamotoi have previously been reported from the Åland Islands but not from the mainland or inner archipelago. The results of the present study suggest an increase in I. ricinus abundance on the island. PMID:26548608

  17. Libraries serving dialogue

    CERN Document Server

    Dupont, Odile

    2014-01-01

    This book based on experiences of libraries serving interreligious dialogue, presents themes like library tools serving dialogue between cultures, collections dialoguing, children and young adults dialoguing beyond borders, story telling as dialog, librarians serving interreligious dialogue.

  18. Multispacer sequence typing relapsing fever Borreliae in Africa.

    Directory of Open Access Journals (Sweden)

    Haitham Elbir

    Full Text Available BACKGROUND: In Africa, relapsing fevers are neglected arthropod-borne infections caused by closely related Borrelia species. They cause mild to deadly undifferentiated fever particularly severe in pregnant women. Lack of a tool to genotype these Borrelia organisms limits knowledge regarding their reservoirs and their epidemiology. METHODOLOGY/PRINCIPAL FINDINGS: Genome sequence analysis of Borrelia crocidurae, Borrelia duttonii and Borrelia recurrentis yielded 5 intergenic spacers scattered between 10 chromosomal genes that were incorporated into a multispacer sequence typing (MST approach. Sequencing these spacers directly from human blood specimens previously found to be infected by B. recurrentis (30 specimens, B. duttonii (17 specimens and B. crocidurae (13 specimens resolved these 60 strains and the 3 type strains into 13 species-specific spacer types in the presence of negative controls. B. crocidurae comprised of 8 spacer types, B. duttonii of 3 spacer types and B. recurrentis of 2 spacer types. CONCLUSIONS/SIGNIFICANCE: Phylogenetic analyses of MST data suggested that B. duttonii, B. crocidurae and B. recurrentis are variants of a unique ancestral Borrelia species. MST proved to be a suitable approach for identifying and genotyping relapsing fever borreliae in Africa. It could be applied to both vectors and clinical specimens.

  19. Improved metod of detection and molecular typing of Borrelia burgdorferi sensu lato in clinical sample by PCR without DNA purification

    Czech Academy of Sciences Publication Activity Database

    Rudenko, Natalia; Golovchenko, Maryna; Němec, J.; Volkaert, J.; Mallátová, N.; Grubhoffer, Libor

    2005-01-01

    Roč. 50, č. 1 (2005), s. 31-39. ISSN 0015-5632 R&D Projects: GA ČR GA524/03/1326 Institutional research plan: CEZ:AV0Z60220518 Keywords : PCR * chelex * amplification Subject RIV: EE - Microbiology, Virology Impact factor: 0.918, year: 2005

  20. Neutrophil Extracellular Traps Entrap and Kill Borrelia burgdorferi Sensu Stricto Spirochetes and Are Not Affected by Ixodes ricinus Tick Saliva

    Czech Academy of Sciences Publication Activity Database

    Menten-Dedoyart, C.; Faccinetto, C.; Golovchenko, Maryna; Dupiereux, I.; Van Lerberghe, P.B.; Dubois, S.; Desmet, C.; Elmoualij, B.; Baron, F.; Rudenko, Natalia; Oury, C.; Heinen, E.; Couvreur, B.

    2012-01-01

    Roč. 189, č. 11 (2012), s. 5393-5401. ISSN 0022-1767 Institutional support: RVO:60077344 Keywords : GROUP-A STREPTOCOCCUS * IMMUNE-RESPONSE * PROTEIN * ESCAPE * INNATE * DEOXYRIBONUCLEASE * EXPRESSION * ALLOWS Subject RIV: EC - Immunology Impact factor: 5.520, year: 2012 http://www.jimmunol.org/content/189/11/5393

  1. Biology of 'Borrelia burgdorferi'-infected and non-infected 'Ixodes ricinus' ticks in the context of climate change

    OpenAIRE

    Herrmann, Coralie; Gern, Lise

    2015-01-01

    La tique Ixodes ricinus est très sensible à la dessiccation et elle est particulièrement exposée à des conditions desséchantes lorsqu’elle quête sur la végétation en attente d’un hôte. Nous avons donc étudié la survie de cette tique à la dessiccation en exposant des nymphes et des adultes à des conditions desséchantes créées en laboratoire à l’aide de solutions salines saturées produisant différentes conditions d’humidité relative à différentes températures. Nous avons observé que lorsque les...

  2. Exploring the association between Morgellons disease and Lyme disease: identification of Borrelia burgdorferi in Morgellons disease patients

    OpenAIRE

    Middelveen, Marianne J; Bandoski, Cheryl; Burke, Jennie; Sapi, Eva; Filush, Katherine R; Wang, Yean; Franco, Agustin; Mayne, Peter J; Stricker, Raphael B.

    2015-01-01

    Background Morgellons disease (MD) is a complex skin disorder characterized by ulcerating lesions that have protruding or embedded filaments. Many clinicians refer to this condition as delusional parasitosis or delusional infestation and consider the filaments to be introduced textile fibers. In contrast, recent studies indicate that MD is a true somatic illness associated with tickborne infection, that the filaments are keratin and collagen in composition and that they result from proliferat...

  3. Diversity and Distribution of Borrelia hermsii

    Science.gov (United States)

    Raffel, Sandra J.; Schrumpf, Merry E.; Porcella, Stephen F.

    2007-01-01

    Borrelia hermsii is the most common cause of tickborne relapsing fever in North America. DNA sequences of the 16S–23S rDNA noncoding intergenic spacer (IGS) region were determined for 37 isolates of this spirochete. These sequences distinguished the 2 genomic groups of B. hermsii identified previously with other loci. Multiple IGS genotypes were identified among isolates from an island, which suggested that birds might play a role in dispersing these spirochetes in nature. In support of this theory, all stages of the tick vector Ornithodoros hermsi fed successfully on birds in the laboratory and advanced in their life cycle. B. hermsii produced a detectable spirochetemia in 1 chicken inoculated subcutaneously. Additional work is warranted to explore the role of birds as enzootic hosts for this relapsing fever spirochete. PMID:17552097

  4. Lyme disease

    Science.gov (United States)

    ... Causes Lyme disease is caused by bacteria called Borrelia burgdorferi ( B burgdorferi ). Blacklegged ticks and other species of ... Names Borreliosis; Bannwarth syndrome Images Lyme disease organism, Borrelia burgdorferi Tick, deer engorged on the skin Lyme disease - ...

  5. A Helpful Serving

    Science.gov (United States)

    Rockower, David

    2006-01-01

    This article briefly describes how a fifth-grade class collaborated with a downtown diner for several months and then actually ran the restaurant for four hours. Through the Chatters Cafe, a local high school cafe that serves as a culinary arts training ground for high school students, fifth graders had the opportunity to prepare and serve dinner…

  6. Pesquisa de anticorpos anti-Borrelia e anti-Babesia em soro de crianças com manifestações clínicas e epidemiologia compatíveis com a doença de Lyme-Simile no Estado de Mato Grosso do Sul Detection of anti-Borrelia and anti-Babesia antibodies in the serum of children with clinical manifestations and compatible epidemiology with Lyme-Like disease in the State of Mato Grosso do Sul

    Directory of Open Access Journals (Sweden)

    Erica Naomi Naka

    2008-04-01

    Full Text Available A ocorrência de manifestações clínicas e laboratoriais semelhantes às encontradas na doença de Lyme e da coinfecção com a babesiose já foi demonstrada em trabalhos anteriores em adultos, porém não existem estudos desta natureza em crianças. OBJETIVO: Caracterizar o perfil clínico-epidemiológico da síndrome de Lyme-Símile em crianças do Estado de Mato Grosso do Sul e avaliar a prevalência dos anticorpos anti-Borrelia burgdorferi e anti-Babesia bovis no soro de pacientes que preencheram os critérios estabelecidos. MÉTODOS: 100 pacientes entre 9 meses e 16 anos de idade foram submetidos à pesquisa dos anticorpos pela técnica de ELISA e a soroprevalência foi comparada com um grupo-controle. RESULTADOS: Positividade para anticorpos anti-B. burgdorferi ocorreu em 27% dos pacientes com suspeita clínica, sendo 17% IgM e 12% IgG. As manifestações articulares ocorreram em 21 pacientes, as manifestações cutâneas em três pacientes e as manifestações neurológicas em três pacientes. A prevalência de anticorpos contra B. burgdorferi nos pacientes do grupo-controle foi de 15% (p The occurence of clinical and laboratory manifestations similar to the Lyme disease and the coinfection with babesiosis was already demonstraded in previous researches with adult patients. However, there are no studies in children. OBJECTIVE: To identify the clinical-epidemiological profile of the Lyme Simile Syndrome in children of the State of Mato Grosso do Sul and to evaluate the prevalence of antibodies against Borrelia burgdorferi and Babesia bovis in the serum of patients that had fulfilled the established criteria. METHODS: One hundred patients (age range: 9 months and 16 years-old were screened for the presence of those antibodies, using ELISA, comparing to a control group. RESULTS: Antibodies against B. burgdoferi were found in 27% of the patients with a clinical picture. The articular manifestations occurred in 21 patients, cutaneous in 3

  7. Transstadial transmission of Borrelia turcica in Hyalomma aegyptium ticks.

    Directory of Open Access Journals (Sweden)

    Zsuzsa Kalmár

    Full Text Available Borrelia turcica comprises the third major group of arthropod-transmitted borreliae and is phylogenetically divergent from other Borrelia groups. The novel group of Borrelia was initially isolated from Hyalomma aegyptium ticks in Turkey and it was recently found in blood and multiple organs of tortoises exported from Jordan to Japan. However, the ecology of these spirochetes and their development in ticks or the vertebrate hosts were not investigated in detail; our aims were to isolate the pathogen and to evaluate the possibility of transstadial transmission of Borrelia turcica by H. aegyptium ticks. Ticks were collected from Testudo graeca tortoises during the summer of 2013 from southeastern Romania. Engorged nymphs were successfully molted to the adult stage. Alive B. turcica was isolated from molted ticks by using Barbour-Stoenner-Kelly (BSK II medium. Four pure cultures of spirochetes were obtained and analyzed by PCR and sequencing. Sequence analysis of glpQ, gyrB and flaB revealed 98%-100% similarities with B. turcica. H. aegyptium ticks collected from T. graeca tortoises were able to pass the infection with B. turcica via transstadial route, suggesting its vectorial capacity.

  8. Failure of Ixodes Ticks To Inherit Borrelia afzelii Infection

    OpenAIRE

    Matuschka, Franz-Rainer; Schinkel, Thomas W.; Klug, Birte; Spielman, Andrew; Richter, Dania

    1998-01-01

    To define conditions promoting inherited infection by Lyme disease spirochetes in Ixodes ticks, we variously infected ticks with Borrelia afzelii and examined their progenies by dark-field microscopy, immunofluorescence, PCR, and serial passage. No episode of inherited infection was evident, regardless of instar or gender infected or frequency of exposure. We suggest that these spirochetes rarely, if ever, are inherited by vector ticks.

  9. Borrelia persica infection in dogs and cats: clinical manifestations, clinicopathological findings and genetic characterization

    OpenAIRE

    Baneth, Gad; Nachum-Biala, Yaarit; Halperin, Tamar; Hershko, Yizhak; Kleinerman, Gabriela; Anug, Yigal; Abdeen, Ziad; Lavy, Eran; Aroch, Itamar; Straubinger, Reinhard K.

    2016-01-01

    Background Relapsing fever (RF) is an acute infectious disease caused by arthropod-borne spirochetes of the genus Borrelia. The disease is characterized by recurrent episodes of fever that concur with spirochetemia. The RF borrelioses include louse-borne RF caused by Borrelia recurrentis and tick-borne endemic RF transmitted by argasid soft ticks and caused by several Borrelia spp. such as B. crocidurae, B. coriaceae, B. duttoni, B. hermsii, B. hispanica and B. persica. Human infection with B...

  10. Serving the Public Interest

    DEFF Research Database (Denmark)

    Markussen, Thomas; Tyran, Jean-Robert

    system is likely to produce good governance. That is, if benevolent candidates are common, if the president has little discretionary power, and if the public sector is effective. We analyze the role of institutions like investigative media and re-election and show that they can improve or further hamper......We present a model of political selection in which voters elect a president from a set of candidates. We assume that some of the candidates are benevolent and that all voters prefer a benevolent president, i.e. a president who serves the public interest. Yet, political selection may fail in our...... model because voters cannot easily tell benevolent from egoistic candidates by observing their pre-election behavior. Egoistic types may strategically imitate benevolent types in the pre-election stage to extract rents once in office. We show that strategic imitation is less likely if the political...

  11. Borrelia recurrentis employs a novel multifunctional surface protein with anti-complement, anti-opsonic and invasive potential to escape innate immunity.

    Directory of Open Access Journals (Sweden)

    Sonja Grosskinsky

    Full Text Available Borrelia recurrentis, the etiologic agent of louse-borne relapsing fever in humans, has evolved strategies, including antigenic variation, to evade immune defence, thereby causing severe diseases with high mortality rates. Here we identify for the first time a multifunctional surface lipoprotein of B. recurrentis, termed HcpA, and demonstrate that it binds human complement regulators, Factor H, CFHR-1, and simultaneously, the host protease plasminogen. Cell surface bound factor H was found to retain its activity and to confer resistance to complement attack. Moreover, ectopic expression of HcpA in a B. burgdorferi B313 strain, deficient in Factor H binding proteins, protected the transformed spirochetes from complement-mediated killing. Furthermore, HcpA-bound plasminogen/plasmin endows B. recurrentis with the potential to resist opsonization and to degrade extracellular matrix components. Together, the present study underscores the high virulence potential of B. recurrentis. The elucidation of the molecular basis underlying the versatile strategies of B. recurrentis to escape innate immunity and to persist in human tissues, including the brain, may help to understand the pathological processes underlying louse-borne relapsing fever.

  12. [Diagnostic aspects of Borrelia-infections in dogs].

    Science.gov (United States)

    Hovius, K E; Houwers, D J

    2007-08-15

    This paper discusses the problem of diagnosing borreliosis (Lyme disease) in dogs. A prospective cohort study in the Kempen district, a known Borrelia focus in The Netherlands, showed that dogs with the presumptive symptoms of borreliosis, episodic malaise and lameness, had significantly higher and longer lasting anti-Borrelia IgG titers than asymptomatic dogs. A small part of these dogs also had antibodies directed against the IR6 (C6) antigen which indicates persistent active Borrelia infection. A few typical case histories are presented. Dogs with episodic malaise and lameness with persistent high IgG titers are suspect of suffering from borreliosis. IR6 antibodies make this diagnosis likely. Initially, such patients should be treated with doxycyclin (10 mg/kg 1dd) for 10 days. If the symptoms recurr within a few months, a longer treatment (eg 6 weeks) should be considered. Bernese mountain dogs were strongly over-represented among the borreliosis patients in the cohort study and most high titered samples among those submitted for--diagnostic--serology appear to come from this breed, which suggests that these dogs have difficulties with clearing this tick-borne infection. PMID:17849909

  13. ServCat Sensitivity Guidelines

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This guide covers sensitivity in ServCat. This document provides technical guidance on how sensitivity fields work in ServCat, and provides suggestions on what...

  14. Tick bite

    Science.gov (United States)

    ... Images Lyme disease, erythema migrans Lyme disease organism, Borrelia burgdorferi Deer ticks Ticks Tick, deer engorged on the skin Lyme disease - Borrelia burgdorferi organism Tick, deer - adult female Deer and dog ...

  15. Lyme borreliosis: insights into tick- / host-borrelia relations

    Czech Academy of Sciences Publication Activity Database

    Grubhoffer, Libor; Golovchenko, Maryna; Vancová, Marie; Zacharovová, Klára; Rudenko, Natalia; Oliver, J. H., Jr.

    2005-01-01

    Roč. 52, č. 4 (2005), s. 279-294. ISSN 0015-5683 R&D Projects: GA ČR GA524/03/1326; GA ČR GA206/03/1323 Grant ostatní: National Institutes of Health(US) R37 AI-24899; Science and Technology Collaboration AVCR-NRC Canada(CZ) Z60220518/58-8500 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia spirochetes * ticks * lectins * glycoproteins Subject RIV: EE - Microbiology, Virology Impact factor: 1.138, year: 2005

  16. Antigenic variation with a twist--the Borrelia story.

    Science.gov (United States)

    Norris, Steven J

    2006-06-01

    A common mechanism of immune evasion in pathogenic bacteria and protozoa is antigenic variation, in which genetic or epigenetic changes result in rapid, sequential shifts in a surface-exposed antigen. In this issue of Molecular Microbiology, Dai et al. provide the most complete description to date of the vlp/vsp antigenic variation system of the relapsing fever spirochaete, Borrelia hermsii. This elaborate, plasmid-encoded system involves an expression site that can acquire either variable large protein (vlp) or variable small protein (vsp) surface lipoprotein genes from 59 different archival copies. The archival vlp and vsp genes are arranged in clusters on at least five different plasmids. Gene conversion occurs through recombination events at upstream homology sequences (UHS) found in each gene copy, and at downstream homology sequences (DHS) found periodically among the vlp/vsp archival genes. Previous studies have shown that antigenic variation in relapsing fever Borrelia not only permits the evasion of host antibody responses, but can also result in changes in neurotropism and other pathogenic properties. The vlsE antigenic variation locus of Lyme disease spirochaetes, although similar in sequence to the relapsing fever vlp genes, has evolved a completely different antigenic variation mechanism involving segmental recombination from a contiguous array of vls silent cassettes. These two systems thus appear to represent divergence from a common precursor followed by functional convergence to create two distinct antigenic variation processes. PMID:16796669

  17. 78 FR 18988 - Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection of...

    Science.gov (United States)

    2013-03-28

    ... Diagnostic Devices for the Detection of Antibodies to Borrelia burgdorferi; Guidance for Industry and Food... Borrelia burgdorferi.'' FDA is issuing this guidance to provide industry and Agency staff with... In Vitro Diagnostic Devices for the Detection of Antibodies to Borrelia burgdorferi'' to the...

  18. 76 FR 570 - Draft Guidance for Industry and Food and Drug Administration Staff; Establishing the Performance...

    Science.gov (United States)

    2011-01-05

    ... Antibodies to Borrelia Burgdorferi; Availability AGENCY: Food and Drug Administration, HHS. ACTION: Notice... Characteristics of In Vitro Diagnostic Devices for the Detection of Antibodies to Borrelia burgdorferi.'' FDA is... detection of antibodies to Borrelia burgdorferi. These devices are used to aid in the diagnosis of...

  19. DNA-based identification and OspC serotyping in cultures of Borrelia burgdorferi s.l. isolated from ticks collected in the Moravia (Czech Republic).

    Science.gov (United States)

    Norek, Adam; Janda, Lubomír; Žákovská, Alena

    2016-06-01

    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). PMID:27232140

  20. 伯氏疏螺旋体与莱姆病研究进展%The Research Advances in Borrelia burgdorferi and Lyme Disease

    Institute of Scientific and Technical Information of China (English)

    宝福凯; 柳爱华

    2007-01-01

    莱姆病是一种新发虫媒传染病,也是一种重要的人兽共患病,其病原体是伯氏疏螺旋体.莱姆病流行范围广,发病率较高,对人类,特别是林区工作者、野外作业人员、旅游者的健康危害大.本文对近年伯氏疏螺旋体与莱姆病研究的某些进展进行简要介绍.

  1. NorAbuMDP glycopeptides and metallochelating nanoliposomes for construction of recombinant vaccines: non-lipidised Borrelia burgdorferi recombinant OspC as an example

    Czech Academy of Sciences Publication Activity Database

    Turánek, J.; Mašek, J.; Raška, M.; Ledvina, Miroslav

    Praha: Ústav organické chemie a biochemie AV ČR, v.v.i, 2011 - (Slaninová, J.; Borovičková, L.). s. 68-68 ISBN 978-80-86241-36-4. [Biologically Active Peptides /12./. 27.04.2011-29.04.2011, Praha] R&D Projects: GA ČR(CZ) GAP304/10/1951; GA AV ČR KAN200520703; GA AV ČR KAN200100801 Institutional research plan: CEZ:AV0Z40550506 Keywords : glycopeptides * synthetic vaccines * structural analysis Subject RIV: CC - Organic Chemistry

  2. Ticks and tick-borne pathogens in South Bohemia (Czech Republic) – Spatial variability in Ixodes ricinus abundance, Borrelia burgdorferi and tick-borne encephalitis virus prevalence

    Czech Academy of Sciences Publication Activity Database

    Hönig, Václav; Švec, P.; Halas, Petr; Vavrušková, Zuzana; Tykalová, Hana; Kilian, Patrik; Vetišková, Vendula; Dorňáková, Veronika; Štěrbová, Jarmila; Šimonová, Zuzana; Erhart, Jan; Štěrba, Ján; Golovchenko, Maryna; Rudenko, Natalia; Grubhoffer, Libor

    2015-01-01

    Roč. 6, č. 5 (2015), s. 559-567. ISSN 1877-959X R&D Projects: GA ČR GD206/09/H026; GA MŠk(CZ) EE2.3.30.0032 Institutional support: RVO:68145535 ; RVO:60077344 Keywords : tick * tick-borne encephalitis * ixodes ricinus * environmental factors * lyme borreliosis Subject RIV: FN - Epidemiology, Contagious Diseases ; Clinical Immunology; EB - Genetics ; Molecular Biology (BC-A) Impact factor: 2.718, year: 2014 http://ac.els-cdn.com/S1877959X15000783/1-s2.0-S1877959X15000783-main.pdf?_tid=8785a702-8de3-11e5-bd28-00000aacb35f&acdnat=1447844587_a91b4e4d404a70fda98b06e4950edb11

  3. 伯氏疏螺旋体外膜蛋白OspA研究进展%Progress in research of Borrelia burgdorferi ourter surface protein A

    Institute of Scientific and Technical Information of China (English)

    崔宇晖; 郭美华; 宝福凯; 柳爱华

    2015-01-01

    莱姆病(Lyme disease,LD)是由伯氏疏螺旋体感染蜱媒传播的人兽共患病.OspA (Outer surface proteinA)是伯氏疏螺旋体主要的外膜蛋白之一.作为主要的免疫原性蛋白,该蛋白有着独特的β-折叠和α-螺旋二级蛋白结构,并且在不同宿主体内有着戏剧性的时空表达规律.OspA具有良好的免疫原性和免疫保护性,因此成为良好的基因疫苗候选者.但OspA株间变异较大,探求流行地区菌株OspA的变异,对疫苗及诊断试剂的研制具有重要意义.本文就该蛋白的结构特点、时空表达特点、异质性、诊断及疫苗的应用等方面归纳总结了近年来的研究成果.

  4. Borrelia burgdorferi s.l. v klíšťatech na ostravských haldách

    Czech Academy of Sciences Publication Activity Database

    Jarošová, V.; Rudolf, Ivo; Halouzka, Jiří; Hubálek, Zdeněk

    2009-01-01

    Roč. 58, č. 2 (2009), s. 90-97. ISSN 1210-7913 R&D Projects: GA AV ČR KJB600930613 Grant ostatní: 6th Framework Programme(XE) GOCE-2003-010284 EDEN Institutional research plan: CEZ:AV0Z60930519 Keywords : Ixodes ricinus * slag heaps * Lyme borreliosis * transmission risk Subject RIV: EE - Microbiology, Virology

  5. A New Borrelia Species Defined by Multilocus Sequence Analysis of Housekeeping Genes▿ †

    OpenAIRE

    Margos, Gabriele; Vollmer, Stephanie A.; Cornet, Muriel; Garnier, Martine; Fingerle, Volker; Wilske, Bettina; Bormane, Antra; Vitorino, Liliana; Collares-Pereira, Margarida; Drancourt, Michel; Kurtenbach, Klaus

    2009-01-01

    Analysis of Lyme borreliosis (LB) spirochetes, using a novel multilocus sequence analysis scheme, revealed that OspA serotype 4 strains (a rodent-associated ecotype) of Borrelia garinii were sufficiently genetically distinct from bird-associated B. garinii strains to deserve species status. We suggest that OspA serotype 4 strains be raised to species status and named Borrelia bavariensis sp. nov. The rooted phylogenetic trees provide novel insights into the evolutionary history of LB spiroche...

  6. Exploring Diversity among Norwegian Borrelia Strains Originating from Ixodes ricinus Ticks

    Directory of Open Access Journals (Sweden)

    Ann-Kristin Tveten

    2014-01-01

    Full Text Available Characterisation of Borrelia strains from Ixodes ricinus ticks is important in the epidemiological surveillance of vector-borne pathogens. Multilocus sequences analysis (MLSA is a molecular genotyping tool with high discriminatory power that has been applied in evolutionary studies and for the characterisation of Borrelia genospecies. MLSA was used to study genetic variations in Borrelia strains isolated from I. ricinus ticks collected from the woodlands in Skodje. The results demonstrate that the 50 Borrelia strains were separated into 36 sequence types (STs that were not previously represented in the MLST database. A distance matrix neighbour-joining tree (bootstrapped 500 iterations showed four deeply branched clusters, and each deeply branched cluster represented one Borrelia genospecies. The mean pairwise genetic differences confirm the genospecies clustering. The combination of alleles separates the Borrelia strains from northwest Norway from the strains in the MLST database, thus identifying new STs. Although a highly divergent B. afzelii population could be expected, the heterogeneity among the B. garinii strains is more unusual. The present study indicates that the circulation of strains between migrating birds and stationary birds in this coastal region may play a role in the evolution of B. garinii strains.

  7. Fundamentals of ServCat

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This training manual for the U.S. Fish and Wildlife Service Catalog (ServCat) provides detailed instructions on searching for records, creating records, and...

  8. Portion Size Versus Serving Size

    Science.gov (United States)

    ... Preschoolers Infographic How to Make a Healthy Home Dietary Recommendations for Healthy Children Top 10 Tips to Help ... for Kids Healthy Foods Under $1 Per Serving Dietary Recommendations for Healthy Children Reading Food Nutrition Labels How ...

  9. NRPC ServCat priorities

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This document lists the Natural Resource Program Centers priority ServCat documents. It is recommended that these documents which include annual narrative reports,...

  10. Natural Infection of Borrelia afzelii in Ixodes sinensis and Its Parasitism Hosts in Forest Areas of Huangshan, Anhui Province%安徽黄山地区中华硬蜱及其宿主感染莱姆病Borrelia afzelii的检测

    Institute of Scientific and Technical Information of China (English)

    孙毅; 许荣满; 葛学峰; 曹务春

    2009-01-01

    A survey was conducted on lxodes sinensis and its parasitism hosts for the infection of Borrelia burgdorferi with PCR and cultivation method. Only one spirochete strain was isolated from 40 I. sinensis collected in Huangshan Mountains. While all blood sample and other I. sinensis ticks fail to get Lyme spirochetes isolation. When detected by 16s rRNA amplification, 6.78% female and 5% male adult I. sinensis ticks yield positive result, two of 92 Chinese hares and one of 16 Chinese muntjaes also appear to be positive with the same DNA band, while all nymph ticks appeared uninfected. However, these DNA amplicon have the same 16S rRNA sequences without any difference. The isolated spirochetes strain obtained from I. sinensis in Huangshan show high similarity to other strains of B. afzelii by phylogeny analysis with 16S rRNA sequences and might belong to Borrelia afzelii species.The finding together with the proved transmission ability, imply that I. sinensis might be the vector tick species in southern China.%利用PCR和病原体分离技术对安徽黄山地区中华硬蜱及其宿主感染莱姆病螺旋体的状况进行了检测,结果从40只黄山林区的中华硬蜱中获得了一株莱姆病螺旋体分离株,寄主动物及其他中华硬蜱均未获得阳性分离结果.以16s rRNA为靶标进行PCR检测,6.78%雌性和5%雄性中华硬蜱发现有阳性感染,2只草兔(92只)和1只麂子(16只)也发现有阳性感染;若蜱和其余宿主动物未发现阳性感染.阳性分离株和扩增获得的168 rRNA序列结果一致,同GenBank中的B.afzelii的系统发育关系最近,提示该分离株螺旋体属于B.afzelii.该结果与前期的实验传播结果提示中华硬蜱是我国南方莱姆病的传播媒介.

  11. Variations in Ixodes ricinus Density and Borrelia Infections Associated with Cattle Introduced into a Woodland in The Netherlands

    NARCIS (Netherlands)

    Gassner, F.; Verbaarschot, P.G.H.; Smallegange, R.C.; Spitzen, J.; Wieren, van S.E.; Takken, W.

    2008-01-01

    The effect of introduced large herbivores on the abundance of Ixodes ricinus ticks and their Borrelia infections was studied in a natural woodland in The Netherlands. Oak and pine plots, either ungrazed or grazed by cattle, were selected. Ticks were collected weekly by blanket dragging. Borrelia inf

  12. Notational Analysis of the Volleyball Serve

    OpenAIRE

    Andrea CIUFFARELLA; Russo, Luca; Masedu, Francesco; Valenti, Marco; Riccardo Edgardo IZZO; Marco DE ANGELIS

    2013-01-01

    The aim of the present study was to investigate the serving techniques in male top level volleyball, especially the tactical and spatial behavior of the servers and receiving opponents focusing on the pros and cons of the different serving techniques. An analysis was made of 4552 serves from 28 matches played during the 2008-2009 regular season of the Italian volleyball male Top League. Serving techniques were categorized into Jump Serve (JS), Float Jump Serve (FJS) and Float Serve (FS), and ...

  13. Prevalence of Borrelia miyamotoi in Ixodes Ticks in Europe and the United States

    Czech Academy of Sciences Publication Activity Database

    Crowder, C. D.; Carolan, H. E.; Rounds, M. A.; Hönig, Václav; Mothes, B.; Haag, H.; Nolte, O.; Luft, B. J.; Grubhoffer, Libor; Ecker, D. J.; Schutzer, S. E.; Eshoo, M. W.

    2014-01-01

    Roč. 20, č. 10 (2014), s. 1678-1682. ISSN 1080-6040 Institutional support: RVO:60077344 Keywords : ionization mass spectrometry * polymerase chain reaction * Lyme disease * burgdorferi * spirochete * infection * vector * meningoencephalitis * identification * pathogens Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 6.751, year: 2014

  14. Borrelia carolinensis sp. nov., a novel species of the Borrelia burgclorferi sensu lato complex isolated from rodents and a tick from the south-eastern USA

    Czech Academy of Sciences Publication Activity Database

    Rudenko, Natalia; Golovchenko, Maryna; Grubhoffer, Libor; Oliver, J. H., Jr.

    2011-01-01

    Roč. 61, č. 2 (2011), 381-383. ISSN 1466-5026 R&D Projects: GA MŠk(CZ) LC06009; GA ČR GA206/09/1782 Institutional research plan: CEZ:AV0Z60220518 Keywords : MULTILOCUS SEQUENCE-ANALYSIS * FRAGMENT-LENGTH-POLYMORPHISM * LYME - DISEASE * RIBOSOMAL-RNA * Borrelia carolinensis sp. nov. * phylogeny Subject RIV: EE - Microbiology, Virology Impact factor: 2.268, year: 2011

  15. A relapsing fever group Borrelia sp. similar to Borrelia lonestari found among wild sika deer (Cervus nippon yesoensis) and Haemaphysalis spp. ticks in Hokkaido, Japan.

    Science.gov (United States)

    Lee, Kyunglee; Takano, Ai; Taylor, Kyle; Sashika, Mariko; Shimozuru, Michito; Konnai, Satoru; Kawabata, Hiroki; Tsubota, Toshio

    2014-10-01

    A relapsing fever Borrelia sp. similar to Borrelia lonestari (herein referred to as B. lonestari-like) was detected from wild sika deer (Cervus nippon yesoensis) and Haemaphysalis ticks in the eastern part of Hokkaido, Japan. The total prevalence of this Borrelia sp. in tested deer blood samples was 10.6% using conventional PCR and real-time PCR. The prevalence was significantly higher in deer fawns compared to adults (21.9% and 9.4%, respectively). Additionally, there was significant regional difference between our two sampling areas, Shiretoko and Shibetsu with 17% and 2.8% prevalence, respectively. Regional differences were also found in tick species collected from field and on deer. In the Shiretoko region, Haemaphysalis spp. were more abundant than Ixodes spp., while in Shibetsu, Ixodes spp. were more abundant. Using real-time PCR analysis, B. lonestari-like was detected from 2 out of 290 adult Haemaphysalis spp. ticks and 4 out of 76 pools of nymphs. This is the first report of a B. lonestari-like organism in Haemaphysalis spp. ticks, and the first phylogenetic analysis of this B. lonestari-like organism in Asia. Based on our results, Haemaphysalis spp. are the most likely candidates to act as a vector for B. lonestari-like; furthermore, regional variation of B. lonestari-like prevalence in sika deer may be dependent on the population distribution of these ticks. PMID:25108784

  16. Tick saliva suppresses IFN signalling in dendritic cells upon Borrelia afzelii infection

    Czech Academy of Sciences Publication Activity Database

    Lieskovská, Jaroslava; Kopecký, Jan

    2012-01-01

    Roč. 34, č. 1 (2012), s. 32-39. ISSN 0141-9838 R&D Projects: GA MŠk(CZ) LC06009 Institutional support: RVO:60077344 Keywords : Borrelia * dendritic cells * interferon signalling * tick saliva Subject RIV: EC - Immunology Impact factor: 2.208, year: 2012

  17. Effect of tick saliva on immune interactions between Borrelia afzelii and murine dendritic cells

    Czech Academy of Sciences Publication Activity Database

    Slámová, M.; Skallová, A.; Páleníková, J.; Kopecký, Jan

    2011-01-01

    Roč. 33, č. 12 (2011), 654-660. ISSN 0141-9838 R&D Projects: GA AV ČR IAA600960811; GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z60220518 Keywords : Borrelia * dendritic cell * immune modulation * Ixodes ricinus Subject RIV: EC - Immunology Impact factor: 2.601, year: 2011

  18. Effect of tick saliva on mechanisms of innate immune response against Borrelia afzelii

    Czech Academy of Sciences Publication Activity Database

    Kýčková, Kateřina; Kopecký, Jan

    2006-01-01

    Roč. 43, č. 4 (2006), s. 1208-1214. ISSN 0022-2585 R&D Projects: GA ČR GA524/05/0811; GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z60220518 Keywords : Ixodes ricinus * Borrelia afzelii * phagocytosis Subject RIV: EC - Immunology Impact factor: 1.950, year: 2006

  19. Tick salivary gland extract-activated transmission of Borrelia afzelii spirochaetes

    Czech Academy of Sciences Publication Activity Database

    Pechová, Jitka; Tresová, Gabriela; Kovář, Lubomír; Kopecký, Jan

    2002-01-01

    Roč. 49, č. 2 (2002), s. 153ů159. ISSN 0015-5683 R&D Projects: GA ČR GA524/02/0901 Institutional research plan: CEZ:AV0Z6022909 Keywords : Ixodes ricinus * saliva ry gland extract * Borrelia afzelii Subject RIV: EC - Immunology Impact factor: 0.515, year: 2002

  20. Borrelia miyamotoi Infection in Patients from Upper Midwestern United States, 2014-2015.

    Science.gov (United States)

    Jobe, Dean A; Lovrich, Steven D; Oldenburg, Darby G; Kowalski, Todd J; Callister, Steven M

    2016-08-01

    We confirmed Borrelia miyamotoi infection in 7 patients who had contracted an illness while near La Crosse, Wisconsin, USA, an area where Ixodes scapularis ticks are endemic. B. miyamatoi infection should now be considered among differential diagnoses for patients from the midwestern United States who have signs and symptoms suggestive of tickborne illness. PMID:27434048

  1. Helical Conformation of Treponema pallidum (Nichols Strain), Treponema paraluis-cuniculi, Treponema denticola, Borrelia turicatae, and Unidentified Oral Spirochetes

    OpenAIRE

    Stepan, Daniel E.; Johnson, Russell C.

    1981-01-01

    Borrelia turicatae (mouse virulent) and Treponema denticola, a small oral treponeme, formed right-handed helices as determined by scanning electron microscopy. Treponema pallidum (Nichols strain), Treponema paraluis-cuniculi, and two unidentified oral spirochetes displayed left-handed helices.

  2. Co-infections with Borrelia species, Anaplasma phagocytophilum and Babesia spp. in patients with tick-borne encephalitis

    OpenAIRE

    Moniuszko, A.; Dunaj, J.; Święcicka, I.; Zambrowski, G.; Chmielewska-Badora, J.; Żukiewicz-Sobczak, W.; Zajkowska, J.; Czupryna, P.; Kondrusik, M.; Grygorczuk, S.; Swierzbinska, R.; Pancewicz, S.

    2014-01-01

    The purpose of this study was evaluation of the prevalence of co-infection with Borrelia species, A. phagocytophilum and Babesia spp. in patients with tick-borne encephalitis (TBE). At total of 110 patients with TBE were included in the study. Serological tests for tick-borne encephalitis virus (TBEV), PCR for Borrelia species, Anaplasma phagocytophilum and Babesia spp., blood smears for A. phagocytophilum and Babesia spp. and BLAST analysis for Babesia spp. were performed. Results showed a s...

  3. ServCat Document Selection Guidelines

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — The ServCat document selection guidelines were developed for selecting appropriate documents to upload into ServCat. When beginning to upload documents into...

  4. Genetic structure of marine Borrelia garinii and population admixture with the terrestrial cycle of Lyme borreliosis

    OpenAIRE

    Gomez-Diaz, E.; Boulinier, T.; Sertour, N.; Cornet, M.; Ferquel, E.; McCoy, Karen

    2011-01-01

    Despite the importance of population structure for the epidemiology of pathogenic bacteria, the spatial and ecological heterogeneity of these populations is often poorly characterized. Here, we investigated the genetic diversity and population structure of the Lyme borreliosis (LB) spirochaete Borrelia garinii in its marine cycle involving colonial seabirds and different host races of the seabird tick Ixodes uriae. Multilocus sequence analyses (MLSA) on eight chromosomal and two plasmid loci ...

  5. Serologic Evidence for Borrelia hermsii Infection in Rodents on Federally Owned Recreational Areas in California

    OpenAIRE

    Fritz, Curtis L; Payne, Jessica R.; Schwan, Tom G.

    2013-01-01

    Tick-borne relapsing fever (TBRF) is endemic in mountainous regions of the western United States. In California, the principal agent is the spirochete Borrelia hermsii, which is transmitted by the argasid tick Ornithodoros hermsi. Humans are at risk of TBRF when infected ticks leave an abandoned rodent nest in quest of a blood meal. Rodents are the primary vertebrate hosts for B. hermsii. Sciurid rodents were collected from 23 sites in California between August, 2006, and September, 2008, and...

  6. Acute Diffuse and Total Alopecia of the Female Scalp Associated with Borrelia-Infection

    OpenAIRE

    Bhardwaj, Ekta K; Trüeb, Ralph Michel

    2015-01-01

    A case of acute diffuse and total alopecia of the female scalp associated with Borrelia-infection (acrodermatitis chronica atrophicans) is presented. Today, acute diffuse and total alopecia of the female scalp is recognized as a distinct variant of alopecia areata (AA) predominantly observed in women. Cases of AA have formerly been reported in association with infections. AA is understood to represent an organ-specific autoimmune disease of the hair follicle. It is conceivable that the antige...

  7. Notational Analysis of the Volleyball Serve

    Directory of Open Access Journals (Sweden)

    Andrea CIUFFARELLA

    2013-12-01

    Full Text Available The aim of the present study was to investigate the serving techniques in male top level volleyball, especially the tactical and spatial behavior of the servers and receiving opponents focusing on the pros and cons of the different serving techniques. An analysis was made of 4552 serves from 28 matches played during the 2008-2009 regular season of the Italian volleyball male Top League. Serving techniques were categorized into Jump Serve (JS, Float Jump Serve (FJS and Float Serve (FS, and for each serve several parameters were recorded: role of the server (Setter - S, Hitter - H, and Middle Blocker - MB, kind of serve (JS, FJS, FS, number of players defending the serve (Defense, difficulty in receiving the serve (RS, evaluation of serve outcome (EV, and defensive court zone where the ball was directed (FZ. The results confirmed the largest use of the JS (69.9%, followed by the FJS (26.9% and the FS (3.3%. There were significant relationships between the serve technique, the EV, the Defense and the FZ where the serves were directed (Chi-Square p = 0.000. The zone absolutely most hit was the posterior/central, followed by the left/posteriors. There were significant differences in the RS difficulty for the JS respect to FJS (p = 0.001 and FS (p = 0.000 and also for the defensive strategies performed: a defense strategy with 2 defenders showed significant and better score during the reception respect to that with 3 (p = 0.000. No statistical relationship was found between the role of the server and other parameters. These results are consistent with previous studies in which JS showed to be the most powerful technique in terms of increasing defensive difficulties but, at the same time, with a fairly high percentage of errors. This issue must be taken into consideration in crucial phases of the game, and the serving techniques must be used strategically. JS can be very useful for increasing the defensive “conflict zones” with the aim to score a

  8. Minority Serving Institutions Reporting System Database

    Data.gov (United States)

    Social Security Administration — The database will be used to track SSA's contributions to Minority Serving Institutions such as Historically Black Colleges and Universities (HBCU), Tribal Colleges...

  9. Statins reduce spirochetal burden and modulate immune responses in the C3H/HeN mouse model of Lyme disease.

    Science.gov (United States)

    Van Laar, Tricia A; Hole, Camaron; Rajasekhar Karna, S L; Miller, Christine L; Reddick, Robert; Wormley, Floyd L; Seshu, J

    2016-06-01

    Lyme disease (LD) is a systemic disorder caused by Borrelia burgdorferi. Lyme spirochetes encode for a functional 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR EC 1.1.1.88) serving as a rate limiting enzyme of the mevalonate pathway that contribute to components critical for cell wall biogenesis. Statins have been shown to inhibit B. burgdorferi in vitro. Using a mouse model of Lyme disease, we found that statins contribute to reducing bacterial burden and altering the murine immune response to favor clearance of spirochetes. PMID:26993029

  10. Detection of Lyme disease spirochetes in the skin of naturally infected wild sika deer (Cervus nippon yesoensis) by PCR.

    Science.gov (United States)

    Kimura, K; Isogai, E; Isogai, H; Kamewaka, Y; Nishikawa, T; Ishii, N; Fujii, N

    1995-04-01

    We demonstrated the presence of Borrelia burgdorferi sensu lato DNA in the skin tissues of naturally infected wild sika deer, using PCR. The risk of transmission of B. burgdorferi sensu lato is recognized in sika deer. PMID:7747979

  11. Detection of Lyme disease spirochetes in the skin of naturally infected wild sika deer (Cervus nippon yesoensis) by PCR.

    OpenAIRE

    Kimura, K.; Isogai, E; Isogai, H; Kamewaka, Y; Nishikawa, T.; Ishii, N; Fujii, N

    1995-01-01

    We demonstrated the presence of Borrelia burgdorferi sensu lato DNA in the skin tissues of naturally infected wild sika deer, using PCR. The risk of transmission of B. burgdorferi sensu lato is recognized in sika deer.

  12. 5 CFR 1203.14 - Serving documents.

    Science.gov (United States)

    2010-01-01

    ... REVIEW OF RULES AND REGULATIONS OF THE OFFICE OF PERSONNEL MANAGEMENT Procedures for Review § 1203.14 Serving documents. (a) Parties. In every case, the person requesting regulation review must serve a copy... business office or home of each party or representative and leaving it with the party or representative,...

  13. Detection of Borrelia-specific 16S rRNA sequence in total RNA extracted from Ixodes ricinus ticks

    Directory of Open Access Journals (Sweden)

    Ž. Radulović

    2010-08-01

    Full Text Available A reverse transcriptase - polymerase chain reaction based assay for Borrelia species detection in ticks was developed. The method was based on amplification of 552 nucleotide bases long sequence of 16S rRNA, targeted by Borrelia specific primers. In the present study, total RNA extracted from Ixodes ricinus ticks was used as template. The results showed higher sensitivity for Borrelia detection as compared to standard dark-field microscopy. Method specificity was confirmed by cloning and sequencing of obtained 552 base pairs long amplicons. Phylogenetic analysis of obtained sequences showed that they belong to B. lusitaniae and B. afzelii genospecies. RT-PCR based method presented in this paper could be very useful as a screening test for detecting pathogen presence, especially when in investigations is required extraction of total RNA from ticks.

  14. "A SURVEY OF PREVALENCE OF ORNITHODORUS TARTAKOVSKYI AND ITS INFECTION RATE WITH BORRELIA LATYCHEVI IN SARAKHS COUNTY,KHORASSAN PROVINCE "

    Directory of Open Access Journals (Sweden)

    N.Piazak

    2000-12-01

    Full Text Available This study was carried out to investigate Borrelia latychevi infection in Omithodorus tartakovskyi in the area surrounding Sarakhs County. Ten percent of the countryside was surveyed for a year and a half from the winter of 1996 to autumn of 1998, in 13 separate field trials. In the nine rural areas inspected, 135 burrows of rodent, terristerid birds and turtles were examined by destruction process. In village areas of Tajan, Sarakhs, Khangiran and Golbibi, respectively, 80%, 26.6%, 16.6% and 13.3% of burrows contained ticks. In the areas of Polkhatoon and Marzdaran no ticks were collected, 200 out of the ticks were studied for infection rate of Borrelia latychevi by direct blood engorgement. Only 30% of the studied ticks were capable of causing Caucasian tick-borne relapsing fever in infant mice, and the rate of infection of Ornithodorus tartakovskyi with Borrelia in Tajan area was 36%.

  15. Biodiversity Information Serving Our Nation (BISON)

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — The USGS Biodiversity Information Serving Our Nation (BISON) project is an online mapping information system consisting of a large collection of species occurrence...

  16. Lyme disease and current aspects of immunization

    OpenAIRE

    Kamradt, Thomas

    2001-01-01

    Lyme disease is a tick-borne multisystem disease that affects primarily the skin, nervous system, heart and joints. At least three species of Borrelia burgdorferi sensu lato, namely Borrelia burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii, can cause the disease. This review will focus mainly on the pathophysiology of Lyme arthritis, the long-term outcome of Lyme disease, and the recently licensed vaccine against Lyme disease.

  17. "Candidatus Borrelia kalaharica" Detected from a Febrile Traveller Returning to Germany from Vacation in Southern Africa.

    Directory of Open Access Journals (Sweden)

    Volker Fingerle

    2016-03-01

    Full Text Available A 26 year-old female patient presented to the Tropical Medicine outpatient unit of the Ludwig Maximilians-University in Munich with febrile illness after returning from Southern Africa, where she contracted a bite by a large mite-like arthropod, most likely a soft-tick. Spirochetes were detected in Giemsa stained blood smears and treatment was started with doxycycline for suspected tick-borne relapsing fever. The patient eventually recovered after developing a slight Jarisch-Herxheimer reaction during therapy. PCR reactions performed from EDTA-blood revealed a 16S rRNA sequence with 99.4% similarity to both, Borrelia duttonii, and B. parkeri. Further sequences obtained from the flagellin gene (flaB demonstrated genetic distances of 0.066 and 0.097 to B. parkeri and B. duttonii, respectively. Fragments of the uvrA gene revealed genetic distance of 0.086 to B. hermsii in genetic analysis and only distant relations with classic Old World relapsing fever species. This revealed the presence of a novel species of tick-borne relapsing fever spirochetes that we propose to name "Candidatus Borrelia kalaharica", as it was contracted from an arthropod bite in the Kalahari Desert belonging to both, Botswana and Namibia, a region where to our knowledge no relapsing fever has been described so far. Interestingly, the novel species shows more homology to New World relapsing fever Borrelia such as B. parkeri or B. hermsii than to known Old World species such as B. duttonii or B. crocidurae.

  18.  The role of complement factor H in the pathogenesis of Borrelia infection

    Directory of Open Access Journals (Sweden)

    Aleksandra Gęca

    2012-07-01

    Full Text Available  Complement factor H (CFH is one of the most important negative regulators of the alternative pathway of the complement system. It is a glycoprotein belonging to the protein H family, which is synthesized mainly in the liver and is composed into a globular protein consisting of 60 amino acid domains in the serum. It shows specificity for C3b molecule of the complement system present in the serum or bound to the cell surface. It inhibits the steady formation of C3 convertase enzymes and the binding of C2 to C4b and factor B to C3b. It accelerates the decomposition of C2a into C4b and the displacement of Bb from C3b.The present paper discusses the composition, properties and functions of the complement factor and the family it belongs to. The paper focuses in particular on its role in the pathogenesis of an infection caused by the spirochetes of the Borrelia genus. Through binding CFH and other related proteins, bacteria of the Borrelia species inhibit the key effect of the alternative pathway of the complement system – the lysis of spirochete cells dependent on the complement’s activation. The mechanism enables pathogens to spread in the host organism and facilitates the evolution of the disease.Discovering the immune mechanisms of the infection caused by the spirochetes of the Borrelia genus may allow for implementing a therapy blocking the binding of complement factor H early enough, apart from the standard treatment of the disease.

  19. Peaceful coexistence amongst Borrelia plasmids: getting by with a little help from their friends?

    OpenAIRE

    Chaconas, George; Norris, Steven J

    2013-01-01

    Borrelia species comprise a unique genus of bacterial pathogens. These organisms contain a segmented genome with up to two dozen plasmids ranging in size from 5kb up to about 200 kb. The plasmids have also been referred to as mini-chromosomes or essential genetic elements, as some of them carry information important for infection of vertebrates or for survival in the tick vector. Most of the plasmids are linear with covalently closed hairpin telomeres and these linear plasmids are in a consta...

  20. Complement-mediated killing of Borrelia garinii--bactericidal activity of wild deer serum.

    Science.gov (United States)

    Isogai, E; Kamewaka, Y; Isogai, H; Kimura, K; Fujii, N; Nishikawa, T

    1994-01-01

    The susceptibility of Borrelia garinii to fresh wild deer sera was determined by incubating strain SIKA2 at 10% serum concentration for 1 hr at 37 C in an in vitro bactericidal assay. Each serum showed bactericidal effects at various levels. The effect was dependent on the concentration of antibody to the spirochetes. Complement was essential in the bactericidal assay because the inactivated deer serum showed greatly decreased activity. Our results suggest that B. garinii is sensitive to deer serum, in the presence of antibody and the bactericidal effect is important for preventing Lyme disease in wild sika deer. PMID:7854216

  1. A notational analysis of elite tennis serve and serve-return strategies on slow surface.

    Science.gov (United States)

    Gillet, Eric; Leroy, David; Thouvarecq, Régis; Stein, Jean-François

    2009-03-01

    A notational analysis of singles events at the French Open Grand Slam tournament was undertaken in 2005 and 2006 to characterize the game patterns and strategies of serve and serve-return and to determine their influence on the point issue on a clay court surface. One hundred sixteen men's singles matches were video analyzed. The flat serve (57.6%), particularly down the "T" location (50.3%), allowed servers to win significantly more points than the topspin (24.1%) and slice serves (18.3%). When the topspin was the first serve strategy, servers kept a high percentage of points won from the serve (52.4%). This strategy was essentially used on the second serve (91.6%) by playing the "T" location in the deuce court and the wide zone in the advantage court. Returns to the central zone allowed receivers to win more points (73.3% on first serve and 65.9% on second serve) than plays to external locations. The results highlight the high impact of the first shots of all opponents on the rally. Even on clay, the slowest court surface, serves and serve-returns remain the strokes that most influence the match results in modern tennis games. PMID:19197212

  2. Plantar pressures in the tennis serve.

    Science.gov (United States)

    Girard, Olivier; Eicher, Frank; Micallef, Jean-Paul; Millet, Grégoire

    2010-06-01

    In-shoe loading patterns were examined in each foot (back and front) separately during two types of tennis serve [first (or flat) and second (or twist) serve] and two service stance styles [foot-up (back foot is moved forward next to front foot for push-off) and foot-back (feet remain at the same relative level)]. Ten competitive tennis players completed five trials for each type of serve and service stance style in random order. Plantar pressure distribution was recorded using Pedar insoles divided into nine areas for analysis. Mean and peak pressures (+15.2%, P forces (+20.2%, P forces were higher (+17.2%, P < 0.05) under the lesser toes. Relative load was higher on the lateral forefoot (+20.4%, P < 0.05) but lower (-32.5%, P < 0.05) on the medial heel of the front foot with foot-up compared with foot-back stance. Using a foot-up stance, loading of the back foot was higher (+31.8%, P < 0.01) under the lateral mid-foot but lower (-29.9%, P < 0.01) under the medial forefoot. The type of serve and the stance style adopted have a significant effect on foot loading. Such findings might help improve mechanical efficiency of the serve. PMID:20496222

  3. Identification of conserved antigens for early serodiagnosis of relapsing fever Borrelia

    Science.gov (United States)

    Lopez, Job E.; Porcella, Stephen F.; Schrumpf, Merry E.; Raffel, Sandra J.; Hammer, Carl H.; Zhao, Ming; Robinson, Mary Ann; Schwan, Tom G.

    2009-01-01

    Borrelia hermsii is a blood-borne pathogen transmitted by the argasid tick Ornithodoros hermsi. Since spirochaete clearance in mice is associated with an IgM-mediated response, an immunoproteomic analysis was used to identify proteins reactive with IgM. We report that IgM from both mice and human patients infected with B. hermsii not only reacted with the previously identified variable membrane proteins but also identified candidate antigens including heat-shock proteins, an adhesin protein, ABC transporter proteins, flagellar proteins, housekeeping proteins, an immune evasion protein, and proteins with unknown function. Furthermore, IgM reactivity to recombinant glycerophosphodiester phosphodiesterase was detected during early spirochaete infection and prior to a detectable IgG response. Lastly, a conserved hypothetical protein was produced in Escherichia coli and tested with immune serum against B. hermsii and Borrelia recurrentis. These results identify a much larger set of immunoreactive proteins, and could help in the early serodiagnosis of this tick-borne infection. PMID:19443544

  4. Strain-specific antibodies reduce co-feeding transmission of the Lyme disease pathogen, Borrelia afzelii.

    Science.gov (United States)

    Jacquet, Maxime; Durand, Jonas; Rais, Olivier; Voordouw, Maarten J

    2016-03-01

    Vector-borne pathogens use a diversity of strategies to evade the vertebrate immune system. Co-feeding transmission is a potential immune evasion strategy because the vector-borne pathogen minimizes the time spent in the vertebrate host. We tested whether the Lyme disease pathogen, Borrelia afzelii, can use co-feeding transmission to escape the acquired immune response in the vertebrate host. We induced a strain-specific, protective antibody response by immunizing mice with one of two variants of OspC (A3 and A10), the highly variable outer surface protein C of Borrelia pathogens. Immunized mice were challenged via tick bite with B. afzelii strains A3 or A10 and infested with larval ticks at days 2 and 34 post-infection to measure co-feeding and systemic transmission respectively. Antibodies against a particular OspC variant significantly reduced co-feeding transmission of the targeted (homologous) strain but not the non-targeted (heterologous) strain. Cross-immunity between OspC antigens had no effect in co-feeding ticks but reduced the spirochaete load twofold in ticks infected via systemic transmission. In summary, OspC-specific antibodies reduced co-feeding transmission of a homologous but not a heterologous strain of B. afzelii. Co-feeding transmission allowed B. afzelii to evade the negative consequences of cross-immunity on the tick spirochaete load. PMID:26411486

  5. Crystal Structure of Neurotropism-Associated Variable Surface Protein 1 (VSP1) of Borrelia Turicatae

    Energy Technology Data Exchange (ETDEWEB)

    Lawson,C.; Yung, B.; Barbour, A.; Zuckert, W.

    2006-01-01

    Vsp surface lipoproteins are serotype-defining antigens of relapsing fever spirochetes that undergo multiphasic antigenic variation to allow bacterial persistence in spite of an immune response. Two isogenic serotypes of Borrelia turicatae strain Oz1 differ in their Vsp sequences and in disease manifestations in infected mice: Vsp1 is associated with the selection of a neurological niche, while Vsp2 is associated with blood and skin infection. We report here crystal structures of the Vsp1 dimer at 2.7 and 2.2 Angstroms. The structures confirm that relapsing fever Vsp proteins share a common helical fold with OspCs of Lyme disease-causing Borrelia. The fold features an inner stem formed by highly conserved N and C termini and an outer 'dome' formed by the variable central residues. Both Vsp1 and OspC structures possess small water-filled cavities, or pockets, that are lined largely by variable residues and are thus highly variable in shape. These features appear to signify tolerance of the Vsp-OspC fold for imperfect packing of residues at its antigenic surface. Structural comparison of Vsp1 with a homology model for Vsp2 suggests that observed differences in disease manifestation may arise in part from distinct differences in electrostatic surface properties; additional predicted positively charged surface patches on Vsp2 compared to Vsp1 may be sufficient to explain the relative propensity of Vsp2 to bind to acidic glycosaminoglycans.

  6. Making a Difference by Serving All Students

    Science.gov (United States)

    Olley, Rivka I.

    2009-01-01

    Randi Brown came to school psychology almost as a family business. Her grandmother was a school psychologist and the first licensed psychologist in the state of New York. Randi graduated with a doctoral degree from Yeshiva University and has served students in Westchester County, New York, for 18 years. She exemplifies the dedication typical of so…

  7. How Finland Serves Gifted and Talented Pupils

    Science.gov (United States)

    Tirri, Kirsi; Kuusisto, Elina

    2013-01-01

    The purpose of this article is to provide an overview of the ways gifted and talented pupils are served in Finland. The trend toward individualism and freedom of choice as well as national policy affecting gifted education are discussed. Empirical research on Finnish teachers' attitudes toward gifted education with respect to the national…

  8. Toss differences between the slice serve and the kick serve in tennis

    OpenAIRE

    Jan Carboch; Vladimír Süss

    2015-01-01

    Background: Pre-contact information of servers' motion is important for receiving players in tennis. Objective: The aim of this study is to examine whether serving players use the same ball toss for kick serve (KS) and slice serve (SS) at two different directions of serves, from the receiver's view. Methods: 10 male right-handed professional tennis players with an average ATP ranking of 533 were videotaped from the receiver's view using a high-speed video camera (200 Hz). Firstly, they...

  9. Acquaintance molestation and youth-serving organizations.

    Science.gov (United States)

    Lanning, Kenneth V; Dietz, Park

    2014-10-01

    This article is based not only on the research literature but also on the extensive field experience of the authors in consulting with investigators, attorneys, and organizations on the prevention, investigation, prosecution, and civil litigation of molestation of children within or in connection with youth-serving organizations. Acquaintance molesters have often pursued careers or sought out paid or volunteer work with organizations through which they can meet children. To address the problem of such offenders, it is necessary for youth-serving organizations to recognize the diversity of sexual activity, the phenomena of "nice-guy" offenders and compliant child victims, and the grooming/seduction process, each of which is reviewed here. The four most important protection practices for organizations are screening; management, and supervision; response to suspicions, allegations, and complaints; and prevention and awareness programs. The authors recommend general approaches to each of these and describe the reasons many organizations resist implementing available preventive measures. PMID:24860081

  10. Serving the fuel cycle: preparing tomorrow's packagings

    International Nuclear Information System (INIS)

    The main fleet of transport packagings serving today the fuel cycle was born more than 20 years ago. Or was it they? The present paper will show that serving the fuel cycle by preparing tomorrow's logistics is actually an on-going process, rather than a rupture. We shall review the great packagings of the fuel cycle: In the front end, the major actors are the UF4, UF6, enriched UF6, UO2 powders, fresh fuel packagings. In the back end of the fuel cycle, we find the dry transport casks of the TN-12, TN-17, TN-13, family and also the Excellox wet flasks. In the waste management, a whole fleet of containers, culminating in the TN Gemini, are available or being created. (author)

  11. Culturally Relevant Practices That "Serve" Students at a Hispanic Serving Institution

    Science.gov (United States)

    Garcia, Gina A.; Okhidoi, Otgonjargal

    2015-01-01

    As institutions not founded to "serve" Latina/o students, Hispanic Serving Institutions (HSIs) must actively change their curricula and programs to meet the needs of their diverse population, including Latina/o, low income, and first generation students. Using a case study approach, including interviews and focus groups, this study…

  12. Toss differences between the slice serve and the kick serve in tennis

    Directory of Open Access Journals (Sweden)

    Jan Carboch

    2015-06-01

    Full Text Available Background: Pre-contact information of servers' motion is important for receiving players in tennis. Objective: The aim of this study is to examine whether serving players use the same ball toss for kick serve (KS and slice serve (SS at two different directions of serves, from the receiver's view. Methods: 10 male right-handed professional tennis players with an average ATP ranking of 533 were videotaped from the receiver's view using a high-speed video camera (200 Hz. Firstly, they served SS and then KS from deuce court. After reaching 3 successful SS and 3 KS to the correct location, the same procedure followed from the ad court. Kinematic analysis was used to obtain the point of ball release, vertical toss peak and racquet-ball contact. Results: Even though the release point was found nearly in the same location, the vertical toss peak of KS was horizontally to the right compared to SS and the point of racquet ball-contact of KS was even more to the right by approximately 30 cm from the receiver's view. Similar findings were obtained from deuce court and ad court. Conclusions: We found differences in the ball toss execution between KS and SS. The serve toss can provide useful information for receiving players. Serving players should use the same toss for each type of serve to hide their intention.

  13. Minority Serving College and University Cost Efficiencies

    Directory of Open Access Journals (Sweden)

    G. Thomas Sav

    2012-01-01

    Full Text Available Problem statement: Higher education minority enrollment growth has far outstripped white non-minority growth in the United States. Minority serving colleges and universities have disproportionately attended to that growth and will continue to play a critical role in providing minority educational opportunities in a knowledge based and globally diverse economy. However, they will face new and challenging budgetary and managerial reforms induced by the global financial crisis. As a result, they will be pressured to operate in the future with greater cost efficiency. Approach: Panel data pertaining to minority serving colleges and universities was used along with stochastic frontier analysis to provide cost inefficiency estimates over a four year academic period. Specification of an inefficiency component contained time varying institutional characteristics and influences, including a public Vs. private ownership control. Results: Minority College and university mean inefficiency was estimated to be approximately 1.24, indicating a 24% operation above the frontier cost. The study found that institutions achieved inefficiency reductions or efficiency gains in 2008-09 compared to 2005-06. The findings suggested that private institutions operated at greater inefficiencies relative to their publicly owned counterparts. However, the private sector laid claim to the most efficient institution, but also the most inefficient one. While the public minority serving colleges showed inefficiency deterioration over time, the findings point to private institution efficiency gains. Conclusion/Recommendations: A literature survey indicated that the study could be the first attempt at providing empirical estimates and subsequent insights into the operating cost efficiencies or inefficiencies of minority serving colleges and universities. The cost inefficiency findings suggested that these institutions did compare favorably in their managerial skills. However, as

  14. Serving the barbarian to preserve the dharma

    OpenAIRE

    Sumit Guha

    2010-01-01

    The last six centuries saw the emergence of major empires that dominated every part of the world. Inevitably then, these imperial formations governed diverse and polyglot peoples. They also developed complex systems of governance that deployed unprecedented numbers of scribes and accountants, often drawn from groups culturally distinct from the military elites. Literati serving Western empires have been much studied of late; but the phenomenon did not originate with them. This article examine...

  15. Serving Diverse Knowledge Systems in Academia

    Directory of Open Access Journals (Sweden)

    William F. Birdsall

    2009-06-01

    Full Text Available Libraries and academic disciplines are experiencing a major transformation to the digital era. A challenge for libraries is to adapt and coordinate their transformation with differing rates and types of changes in teaching, research, and scholarly communication among the disciplines they serve. This paper argues libraries need to acknowledge the diversity of knowledge systems and adopt a strategy that requires collaboration between libraries and multiple communities of knowing in the development and provision of heterogeneous services.

  16. DO ACCOUNTING AND FINANCE TOOLS SERVE GOVERNANCE?

    OpenAIRE

    Mohamed Ariff; J. Ratnatunga

    2008-01-01

    A brief review of recent literature on corporate governance is provided, which is then concluded with a proposed corporate governance framework as a starting point for further development. We propose that it is stakeholder concentration that determines the quality of corporate governance. Next objective of this paper is the more ambitious one of addressing the role of accounting and finance disciplines to serve corporate governance. We test empirically if the use of some accounting and financ...

  17. 莱姆病螺旋体基因型SPR鉴定技术的研究%Studies on the Detection of Genotype of Lyme Disease Borrelia Burgdorferi by SPR

    Institute of Scientific and Technical Information of China (English)

    于东冬; 刘岩; 张磊; 周建光

    2009-01-01

    采用自行设计组装的一套新型表面等离子体子共振(SPR)传感器, 并通过测量计算免疫反应动力学参数--解离常数KD, 对吉林地区常见的莱姆病螺旋体基因型(B.afzelii型和B.garinii型)进行了鉴定研究. 根据实验动物和实际病患血清的鉴定结果可以初步证明, 采用波长型SPR传感器鉴定莱姆病螺旋体基因型具有操作简单、节省时间以及仪器易于小型化、便于推广等特点.

  18. 分析伯氏疏螺旋体密码子使用的倾向性%A nonlinear mapping for codon usage bias studies in borrelia burgdorferi genome

    Institute of Scientific and Technical Information of China (English)

    韩志娴; 王永宏; 展永

    2005-01-01

    本文采用非线性映射的方法分析伯氏疏螺旋体前导链和后随链上的基因结构,发现基因分布存在明显的差异,同义密码子的使用亦具有明显的倾向性.此外,高表达基因分布具有不对称性,其同义密码子使用与其它基因亦有不同,这表明原核生物基因组复制起始点两侧的碱基分布及翻译机制均影响基因的密码子使用.

  19. Multiplex assay (Mikrogen recomBead) for detection of serum IgG and IgM antibodies to 13 recombinant antigens of Borrelia burgdorferi sensu lato in patients with neuroborreliosis

    DEFF Research Database (Denmark)

    Dessau, Ram Benny; Møller, Jens K.; Kolmos, Birte;

    2015-01-01

    with Lyme neuroborreliosis (LNB), 218 Danish blood donor controls, a set of 61 Swedish patients with LNB and 139 Swedish non-LNB patients investigated for suspected LNB were used. There are four parts developed in this study: a characterization of the sero-epidemiological antibody-response pattern, the...

  20. The crystal structures of two salivary cystatins from the tick Ixodes scapularis and the effect of these inhibitors on the establishment of Borrelia burgdorferi infection in a murine model

    Czech Academy of Sciences Publication Activity Database

    Kotsyfakis, Michalis; Horká, Helena; Salát, Jiří; Andersen, J. F.

    2010-01-01

    Roč. 77, č. 2 (2010), s. 456-470. ISSN 0950-382X R&D Projects: GA AV ČR KJB500960702; GA AV ČR IAA600960811 Institutional research plan: CEZ:AV0Z60220518 Keywords : INVARIANT CHAIN FRAGMENT * EGG-WHITE CYSTATIN * CATHEPSIN-L * CYSTEINE PROTEINASES * SIALOSTATIN-L * ENDOPEPTIDASE Subject RIV: EC - Immunology Impact factor: 4.819, year: 2010