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Sample records for borrelia burgdorferi hbb

  1. Asymptomatic infection with Borrelia burgdorferi.

    Science.gov (United States)

    Steere, Allen C; Sikand, Vijay K; Schoen, Robert T; Nowakowski, John

    2003-08-15

    The natural history of asymptomatic seroconversion to Borrelia burgdorferi has been unclear. We report here, on the basis of a post hoc assessment, the frequency and outcome of asymptomatic seroconversion to B. burgdorferi in participants of a large Lyme disease vaccine trial. We show that infection with B. burgdorferi may be asymptomatic but that asymptomatic infection is unusual in the United States.

  2. 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...

  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. 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

  5. Recombinant constructs of Borrelia burgdorferi

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    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.

  6. 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...

  7. 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.

  8. The lipid raft proteome of Borrelia burgdorferi.

    Science.gov (United States)

    Toledo, Alvaro; Pérez, Alberto; Coleman, James L; Benach, Jorge L

    2015-11-01

    Eukaryotic lipid rafts are membrane microdomains that have significant amounts of cholesterol and a selective set of proteins that have been associated with multiple biological functions. The Lyme disease agent, Borrelia burgdorferi, is one of an increasing number of bacterial pathogens that incorporates cholesterol onto its membrane, and form cholesterol glycolipid domains that possess all the hallmarks of eukaryotic lipid rafts. In this study, we isolated lipid rafts from cultured B. burgdorferi as a detergent resistant membrane (DRM) fraction on density gradients, and characterized those molecules that partitioned exclusively or are highly enriched in these domains. Cholesterol glycolipids, the previously known raft-associated lipoproteins OspA and OpsB, and cholera toxin partitioned into the lipid rafts fraction indicating compatibility with components of the DRM. The proteome of lipid rafts was analyzed by a combination of LC-MS/MS or MudPIT. Identified proteins were analyzed in silico for parameters that included localization, isoelectric point, molecular mass and biological function. The proteome provided a consistent pattern of lipoproteins, proteases and their substrates, sensing molecules and prokaryotic homologs of eukaryotic lipid rafts. This study provides the first analysis of a prokaryotic lipid raft and has relevance for the biology of Borrelia, other pathogenic bacteria, as well as for the evolution of these structures. All MS data have been deposited in the ProteomeXchange with identifier PXD002365 (http://proteomecentral.proteomexchange.org/dataset/PXD002365).

  9. 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.

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

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    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.

  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.

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    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. 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.

  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

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    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. Detection of Borrelia burgdorferi and Borrelia lonestari in birds in Tennessee.

    Science.gov (United States)

    Jordan, B E; Onks, K R; Hamilton, S W; Hayslette, S E; Wright, S M

    2009-01-01

    Lyme disease in the United States is caused by the bacterial spirochete Borrelia burgdorferi s.s. (Johnson, Schmid, Hyde, Steigerwalt, and Brenner), which is transmitted by tick vectors Ixodes scapularis (Say) and I. pacificus (Cooley and Kohls). Borrelia lonestari, transmitted by the tick Amblyomma americanum L., may be associated with a related syndrome, southern tick-associated rash illness (STARI). Borrelia lonestari sequences, reported primarily in the southeastern states, have also been detected in ticks in northern states. It has been suggested that migratory birds may have a role in the spread of Lyme disease spirochetes. This study evaluated both migratory waterfowl and nonmigratory wild turkeys (Meleagris gallopavo silvestris, Eastern wild turkey) for B. burgdorferi and B. lonestari DNA sequences. A total of 389 avian blood samples (163 migratory birds representing six species, 125 wild turkeys harvested in habitats shared with migratory birds, 101 wild turkeys residing more distant from migratory flyways) were extracted, amplified, and probed to determine Borrelia presence and species identity. Ninety-one samples were positive for Borrelia spp. Among migratory birds and turkeys collected near migration routes, B. burgdorferi predominated. Among turkeys residing further away from flyways, detection of B. lonestari was more common. All A. americanum ticks collected from these areas were negative for Borrelia DNA; no I. scapularis were found. To our knowledge, this represents the first documentation of B. lonestari among any birds.

  1. Monoinfections caused by Borrelia burgdorferi and Borrelia burgdorferi / Anaplasma phagocytophilum co-infections in forestry workers and farmers

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    Małgorzata Tokarska-Rodak

    2015-10-01

    Full Text Available Background: The presence of co-infections induced by tick-borne pathogens in humans is an important epidemiological phenomenon. This issue has attracted growing attention of doctors and people working under conditions of an increased risk of being exposed to tick bites. Material and Methods: The research group consisted of 93 individuals with current anti-immunoglobulin M/G (IgM/ IgG Borrelia burgdorferi or IgG anti-Anaplasma phagocytophilum. The respondents were identified during the screening survey in a group of farmers and foresters occupationally exposed to tick bites. The aim of the work was to analyse the frequency of antibodies to specific antigens of B. burgdorferi and the levels of cytokines in forestry workers and farmers with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infections. Statistical analysis was performed using the Chi2, Mann-Whitney U and Kruskal-Wallis tests. Results: There is a stronger generation of IgG antibodies to B. burgdorferi antigens in patients with B. burgdorferi / A. phagocytophilum co-infections, such as variable major protein-like sequence expressed (VlsE (p < 0.05, p19 (p < 0.02, p17 (p < 0.05 and complement regulator-acquiring surface protein 3 (CRASP3 (p < 0.02 compared to persons with B. burgdorferi monoinfections. The discrepancies in the synthesis of cytokines interleukin 6 (IL-6, IL-10, and tumor necrosis factor α (TNF-α have not been found in persons with B. burgdorferi monoinfections and B. burgdorferi / A. phagocytophilum co-infection. Conclusions: The immune response directed against B. burgdorferi is stronger in patients co-infected with B. burgdorferi and A. phagocytophilum than in those with monoinfection. Med Pr 2015;66(5:645–651

  2. 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

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

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    Gorana Veinović

    Full Text Available 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 (P0.05, respectively. Inoculum had statistically significant influence on growth of all three Borrelia species at all tested temperatures except at 4°C.

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

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    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...

  5. Minimal-Change Disease Secondary to Borrelia burgdorferi Infection

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    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. ...

  6. Borrelia burgdorferi Spirochetes Induce Mast Cell Activation and Cytokine Release

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    Talkington, Jeffrey; Nickell, Steven P.

    1999-01-01

    The Lyme disease spirochete, Borrelia burgdorferi, is introduced into human hosts via tick bites. Among the cell types present in the skin which may initially contact spirochetes are mast cells. Since spirochetes are known to activate a variety of cell types in vitro, we tested whether B. burgdorferi spirochetes could activate mast cells. We report here that freshly isolated rat peritoneal mast cells or mouse MC/9 mast cells cultured in vitro with live or freeze-thawed B. burgdorferi spirochetes undergo low but detectable degranulation, as measured by [5-3H] hydroxytryptamine release, and they synthesize and secrete the proinflammatory cytokine tumor necrosis factor alpha (TNF-α). In contrast to findings in previous studies, where B. burgdorferi-associated activity was shown to be dependent upon protein lipidation, mast cell TNF-α release was not induced by either lipidated or unlipidated recombinant OspA. This activity was additionally shown to be protease sensitive and surface expressed. Finally, comparisons of TNF-α-inducing activity in known low-, intermediate-, and high-passage B. burgdorferi B31 isolates demonstrated passage-dependent loss of activity, indicating that the activity is probably plasmid encoded. These findings document the presence in low-passage B. burgdorferi spirochetes of a novel lipidation-independent activity capable of inducing cytokine release from host cells. PMID:10024550

  7. Proteome Analysis of Borrelia burgdorferi Response to Environmental Change

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    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.

  8. 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.

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

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    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.

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

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    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.'

  11. Lipid exchange between Borrelia burgdorferi and host cells.

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    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.

  12. [Heterogeneity of the gene P83/100 of Borrelia borrelia burgdorferi sensu lato complex].

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    Fomenko, N V; Sabitova, Iu B; Khasnatinov, M A; Gol'tsova, N A; Danchinova, G A; Bataa, Zh; Ambed, D; Stronin, O V

    2007-01-01

    The 35 full-length Borrelia burgdorferi sensu lato complex a83/100 gene nucleotide sequences were determined. High level of homology was observed in the nucleotide sequences corresponding to the strains and isolates of Borrelia fzelii. The analysis of the nucleotide sequences revealed two groups of Borrelia garinii. The most variable p83/100 gene region containing species-typical insertions and deletions was demonstrated to be included into the region where the antigenic determinants of protein were encoded. According to the data obtained in this work, the modification of the P83/100 protein structure and immunological properties could be suggested to exist even within species. The results of this work could be used for receiving recombinant P83/100 proteins useful for diagnostic applications.

  13. 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,...

  14. 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...

  15. [Criteria for evaluation of immunoblots using Borrelia afzelii, Borrelia garinii and Borrelia burgdorferi sensu stricto for diagnosis of Lyme borreliosis].

    Science.gov (United States)

    Honegr, K; Havlasová, J; Gebouský, P; Dostál, V; Pellantová, V; Skrabková, Z; Hulínská, D

    2001-11-01

    The immunoblot was prepared from genotypes Borrelia afzelii (KC 90), Borrelia garinii (M 192) and Borrelia burgdorferi sensu stricto (B 31). Sera of 63 patients with different forms of Lyme borreliosis were examined and 40 healthy donors in the endemic area of the disease. In class IgM in the group of patients significantly more frequently antibodies against OspC, p39, p41 B. afzelii, p39, p41, p66, p83 B. garinii and OspC1, OspA, B. burgdorferi sensu stricto were found. In class IgG there were antibodies against p39, p41, p93 B. afzelii, p14, p41, p93 B. garinii and OspA, OspC p93 B. burgdorferi sensu lato. Based on the assembled results by means of discrimination analysis and logistic regression the most suitable combinations of antigens for evaluation of immunoblots in different genotypes were determined. Furthermore evaluation was suggested using a combination of antigens of several genotypes which led to an increased sensitivity and specificity of the immunoblot. Tables were prepared for easier evaluation of newly examined sera samples.

  16. 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

  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

    Directory of Open Access Journals (Sweden)

    Elizabeta S. Ristanovic

    2007-03-01

    Full Text Available 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 A1, A2 e M1, isoladas de Ixodes ricinus, pertencem à Borrelia burgdorferi sensu stricto, enquanto a cepa K1, isolada de Apodemus flavocollis é uma mistura de Borrelia afzelii e B. burgdorferi s.s.

  18. 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...

  19. 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...

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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...

  7. 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...

  8. Antibodies to Rickettsia spp. and Borrelia burgdorferi in Spanish Wild Red Foxes (Vulpes vulpes).

    Science.gov (United States)

    Lledó, Lourdes; Serrano, José Luis; Isabel Gegúndez, María; Giménez-Pardo, Consuelo; Saz, José Vicente

    2016-01-01

    We examined 314 red foxes (Vulpes vulpes) from the province of Soria, Spain, for Rickettsia typhi, Rickettsia slovaca, and Borrelia burgdorferi infection. Immunofluorescence assays showed 1.9% had antibodies to R. typhi, 6.7% had antibodies to R. slovaca, and 8.3% had antibodies to B. burgdorferi. Serostatus was not correlated with sex or age. Because red foxes can be infected by Rickettsiae and B. burgdorferi, presence of red foxes may be and indicator for the presence of these pathogens.

  9. 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...

  10. 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...

  11. Patterns and regulation of ribosomal RNA transcription in Borrelia burgdorferi

    Directory of Open Access Journals (Sweden)

    Schwartz Ira

    2011-01-01

    Full Text Available Abstract Background Borrelia burgdorferi contains one 16S and two tandem sets of 23S-5S ribosomal (r RNA genes whose patterns of transcription and regulation are unknown but are likely to be critical for survival and persistence in its hosts. Results RT-PCR of B. burgdorferi N40 and B31 revealed three rRNA region transcripts: 16S rRNA-alanine transfer RNA (tRNAAla; tRNAIle; and both sets of 23S-5S rRNA. At 34°C, there were no differences in growth rate or in accumulation of total protein, DNA and RNA in B31 cultured in Barbour-Stoenner-Kelly (BSK-H whether rabbit serum was present or not. At 23°C, B31 grew more slowly in serum-containing BSK-H than at 34°C. DNA per cell was higher in cells in exponential as compared to stationary phase at either temperature; protein per cell was similar at both temperatures in both phases. Similar amounts of rRNA were produced in exponential phase at both temperatures, and rRNA was down-regulated in stationary phase at either temperature. Interestingly, a relBbu deletion mutant unable to generate (pppGpp did not down-regulate rRNA at transition to stationary phase in serum-containing BSK-H at 34°C, similar to the relaxed phenotype of E. coli relA mutants. Conclusions We conclude that rRNA transcription in B. burgdorferi is complex and regulated both by growth phase and by the stringent response but not by temperature-modulated growth rate.

  12. 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.

  13. 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.

  14. 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).

  15. 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

  16. 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.

  17. 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...

  18. 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

  19. 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...

  20. 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

  1. Borrelia burgdorferi Aggrecanase Activity: More Evidence for Persistent Infection in Lyme Disease.

    Directory of Open Access Journals (Sweden)

    Raphael B. Stricker

    2013-08-01

    Full Text Available 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.

  2. 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...

  3. 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.

  4. 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...

  5. 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.

  6. 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.

  7. 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...

  8. 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.

  9. 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

  10. The urokinase receptor (uPAR facilitates clearance of Borrelia burgdorferi.

    Directory of Open Access Journals (Sweden)

    Joppe W R Hovius

    2009-05-01

    Full Text Available 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, dependently or independently of ligation to uPA, directly affect leukocyte function. We here demonstrate that uPAR is upregulated on murine and human leukocytes upon exposure to B. burgdorferi both in vitro as well as in vivo. Notably, B. burgdorferi-inoculated C57BL/6 uPAR knock-out mice harbored significantly higher Borrelia numbers compared to WT controls. This was associated with impaired phagocytotic capacity of B. burgdorferi by uPAR knock-out leukocytes in vitro. B. burgdorferi numbers in vivo, and phagocytotic capacity in vitro, were unaltered in uPA, tPA (low fibrinolytic activity and PAI-1 (high fibrinolytic activity knock-out mice compared to WT controls. Strikingly, in uPAR knock-out mice partially backcrossed to a B. burgdorferi susceptible C3H/HeN background, higher B. burgdorferi numbers were associated with more severe carditis and increased local TLR2 and IL-1beta mRNA expression. In conclusion, in B. burgdorferi infection, uPAR is required for phagocytosis and adequate eradication of the spirochete from the heart by a mechanism that is independent of binding of uPAR to uPA or its role in the fibrinolytic system.

  11. 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.

  12. Autophagy Modulates Borrelia burgdorferi-induced Production of Interleukin-1beta (IL-1beta)

    NARCIS (Netherlands)

    Buffen, K.; Oosting, M.; Mennens, S.; Anand, P.K.; Plantinga, T.S.; Sturm, P.D.J.; Veerdonk, F.L. van de; Meer, J.W. van der; Xavier, R.J.; Kanneganti, T.D.; Netea, M.G.; Joosten, L.A.B.

    2013-01-01

    Borrelia burgdorferi sensu lato is the causative agent of Lyme disease. Recent studies have shown that recognition of the spirochete is mediated by TLR2 and NOD2. The latter receptor has been associated with the induction of the intracellular degradation process called autophagy. The present study d

  13. 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

  14. Comparison of detection of Borrelia burgdorferi DNA and anti-Borrelia burgdorferi antibodies in patients with erythema migrans in north-eastern Poland

    Science.gov (United States)

    Dunaj, Justyna; Zajkowska, Joanna; Czupryna, Piotr; Świerzbińska, Renata; Guziejko, Katarzyna; Aleksiejczuk, Piotr; Barry, Gerald; Kondrusik, Maciej; Pancewicz, Sławomir

    2015-01-01

    Introduction Diagnostic methods in erythema migrans are still not standardized. Aim To evaluate the frequency of Borrelia burgdorferi s.l. DNA presence in patients with erythema migrans (EM); to assess the polymerase chain reaction (PCR) procedure for detecting B. burgdorferi s.l. DNA in patients with the skin form of Lyme borreliosis; and to compare the results of the PCR-based method with the traditional ELISA method. Material and methods Skin biopsy and blood samples from 93 patients with EM were examined for B. burgdorferi s.l. DNA detection (PCR). Seventy-one of these patients were examined for the presence of anti-B. burgdorferi s.l. antibodies (ELISA). Results Borrelia burgdorferi s.l. DNA was detected in 48% of the skin biopsy specimens and in 2% of blood samples. Only 1 patient was PCR positive in both blood and skin samples. Seventy percent of patients whose PCR results were positive were bitten by a tick less than 14 days before. IgM anti-B. burgdorferi s.l – specific antibodies were present in the serum of 35% of patients and IgG antibodies – in 30% of patients. Seventeen percent were positive in both IgM and IgG. Conclusions Polymerase chain reaction of skin biopsy specimens seems to be currently the most sensitive and specific test for the diagnosis of patients with EM, especially in patients with a short duration of the disease (< 14 days) but still its effectiveness is much lower than expected. Polymerase chain reaction of blood samples cannot be recommended at the present time for the routine diagnostic of patients with EM. PMID:25821421

  15. 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

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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...

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  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. Molecular characterization of the p100 gene of Borrelia burgdorferi strain PKo.

    Science.gov (United States)

    Jauris-Heipke, S; Fuchs, R; Hofmann, A; Lottspeich, F; Preac-Mursic, V; Soutschek, E; Will, G; Wilske, B

    1993-12-01

    The p100 gene coding for the p100 protein of Borrelia burgdorferi strain PKo has been cloned, sequenced and expressed in Escherichia coli. An open reading frame including upstream and downstream sequences with potential translation and transcription signals could be identified. The reading frame consists of 1989 nucleotides corresponding to a protein of 663 amino acids and a calculated molecular mass of 75.8 kDa. The protein has a leader peptide and is processed without modification at the N-terminus. A high percentage of amino acid sequence identity could be found to the high-molecular mass protein p83/p93 of B. burgdorferi strain B31.

  8. 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.

  9. 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.

  10. 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....

  11. 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% ...

  12. 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...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. Evidence that Borrelia burgdorferi immunodominant proteins p100, p94 and p83 are identical.

    Science.gov (United States)

    Ditton, H J; Neuss, M; Zöller, L

    1992-07-15

    Recently there have been reports on high-molecular mass components of Borrelia burgdorferi, namely the p100, p94 and p83, which claimed these proteins to be specific marker antigens for the serodiagnosis of late Lyme borreliosis. The nucleotide sequences of the p100 and p83 have been published. The alignment of the deduced N-terminal amino acid sequences with the N-terminal sequence of the p94 now provides evidence that all three proteins are identical.

  2. Analysis of Borrelia burgdorferi surface proteins as determinants in establishing host cell interactions

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    Virginia L Schmit

    2011-07-01

    Full Text Available Borrelia burgdorferi infection causes Lyme borreliosis in humans, a condition which can involve a systemic spread of the organism to colonize various tissues and organs. If the infection is left untreated by antimicrobials, it can lead to manifestations including, arthritis, carditis, and/or neurological problems. Identification and characterization of B. burgdorferi outer membrane proteins that facilitate cellular attachment and invasion to establish infection continue to be investigated. In this study, we sought to further define putative cell binding properties of surface-exposed B. burgdorferi proteins by observing whether cellular adherence could be blocked by antibodies. B. burgdorferi mixed separately with monoclonal antibodies against outer surface protein (Osp A, OspC, decorin-binding protein (Dbp A, BBA64, and RevA antigens were incubated with human umbilical vein endothelial cells (HUVEC and human neuroglial cells (H4. B. burgdorferi treated with anti-OspA, -DbpA, and –BBA64 monoclonal antibodies showed a significant decrease in cellular association compared to controls, whereas B. burgdorferi treated with anti-OspC and anti-RevA showed no reduction in cellular attachment. Additionally, temporal transcriptional analyses revealed upregulated expression of bba64, ospA, and dbpA during coincubation with cells. Together, the data provide evidence that OspA, DbpA, and BBA64 function in host cell adherence and infection mechanisms.

  3. A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection

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    Wei Liu

    2016-08-01

    Full Text Available A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA was established for Borrelia burgdorferi (B. burgdorferi detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients’ serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions.

  4. A Novel Isothermal Assay of Borrelia burgdorferi by Recombinase Polymerase Amplification with Lateral Flow Detection.

    Science.gov (United States)

    Liu, Wei; Liu, Hui-Xin; Zhang, Lin; Hou, Xue-Xia; Wan, Kang-Lin; Hao, Qin

    2016-01-01

    A novel isothermal detection for recombinase polymerase amplification with lateral flow (LF-RPA) was established for Borrelia burgdorferi (B. burgdorferi) detection in this study. This assay with high sensitivity and specificity can get a visible result without any additional equipment in 30 min. We designed a pair of primers according to recA gene of B. burgdorferi strains and a methodology evaluation was performed. The results showed that the RPA assay based on the recA gene was successfully applied in B. burgdorferi detection, and its specific amplification was only achieved from the genomic DNA of B. burgdorferi. The detection limit of the new assay was about 25 copies of the B. burgdorferi genomic DNA. Twenty Lyme borreliosis patients' serum samples were detected by LF-RPA assay, real-time qPCR and nested-PCR. Results showed the LF-RPA assay is more effective than nested-PCR for its shorter reaction time and considerably higher detection rate. This method is of great value in clinical rapid detection for Lyme borreliosis. Using the RPA assay might be a megatrend for DNA detection in clinics and endemic regions. PMID:27527151

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

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    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. 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

  7. 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

  8. 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.

  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. BB0744 Affects Tissue Tropism and Spatial Distribution of Borrelia burgdorferi.

    Science.gov (United States)

    Wager, Beau; Shaw, Dana K; Groshong, Ashley M; Blevins, Jon S; Skare, Jon T

    2015-09-01

    Borrelia burgdorferi, the etiologic agent of Lyme disease, produces a variety of proteins that promote survival and colonization in both the Ixodes species vector and various mammalian hosts. We initially identified BB0744 (also known as p83/100) by screening for B. burgdorferi strain B31 proteins that bind to α1β1 integrin and hypothesized that, given the presence of a signal peptide, BB0744 may be a surface-exposed protein. In contrast to this expectation, localization studies suggested that BB0744 resides in the periplasm. Despite its subsurface location, we were interested in testing whether BB0744 is required for borrelial pathogenesis. To this end, a bb0744 deletion was isolated in a B. burgdorferi strain B31 infectious background, complemented, and queried for the role of BB0744 following experimental infection. A combination of bioluminescent imaging, cultivation of infected tissues, and quantitative PCR (qPCR) demonstrated that Δbb0744 mutant B. burgdorferi bacteria were attenuated in the ability to colonize heart tissue, as well as skin locations distal to the site of infection. Furthermore, qPCR indicated a significantly reduced spirochetal load in distal skin and joint tissue infected with Δbb0744 mutant B. burgdorferi. Complementation with bb0744 restored infectivity, indicating that the defect seen in Δbb0744 mutant B. burgdorferi was due to the loss of BB0744. Taken together, these results suggest that BB0744 is necessary for tissue tropism, particularly in heart tissue, alters the ability of B. burgdorferi to disseminate efficiently, or both. Additional studies are warranted to address the mechanism employed by BB0744 that alters the pathogenic potential of B. burgdorferi.

  11. 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...

  12. 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.

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

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    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.

  14. Borrelia burgdorferi erp genes are expressed at different levels within tissues of chronically infected mammalian hosts.

    Science.gov (United States)

    Miller, Jennifer C; Stevenson, Brian

    2006-05-01

    The spirochete Borrelia burgdorferi is the causative agent of Lyme disease and is transmitted to humans and other vertebrate hosts through the bites of ixodid ticks. B. burgdorferi Erp (OspE-F related lipoprotein) family members are encoded on members of the 32 kb circular plasmid-like prophage family (cp32s). Many Erp proteins serve as receptors for the complement inhibitory factor H molecules of numerous vertebrate hosts, providing one mechanism by which the bacteria potentially evade the innate immune system. Indirect immunofluorescence analyses (IFA) have demonstrated that Erp expression is temporally regulated throughout the mammal-tick infectious cycle, indicating that Erp proteins perform an important role (or even roles) during mammalian infection. However, it was not previously known whether Erp proteins are continually produced by B. burgdorferi throughout the course of mammalian infection. To address this issue, quantitative RT-PCR (q-RT-PCR) was utilized to assess erp transcription levels by bacteria within numerous different tissues of both mice and non-human primates (NHPs) chronically infected with B. burgdorferi. Q-RT-PCR results obtained using both animal models indicated that while the majority of erp genes were detectably transcribed during chronic infection, differences in expression levels were noted. These data strongly suggest that Erp proteins contribute to B. burgdorferi persistence within chronically infected host tissues, perhaps by protecting the bacteria from complement-mediated killing. PMID:16530008

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

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    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

  17. 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.

  18. Occurrence of Borrelia burgdorferi s.l. in different genera of mosquitoes (Culicidae) in Central Europe.

    Science.gov (United States)

    Melaun, Christian; Zotzmann, Sina; Santaella, Vanesa Garcia; Werblow, Antje; Zumkowski-Xylander, Helga; Kraiczy, Peter; Klimpel, Sven

    2016-03-01

    Lyme disease or Lyme borreliosis is a vector-borne infectious disease caused by spirochetes of the Borrelia burgdorferi sensu lato complex. Some stages of the borrelial transmission cycle in ticks (transstadial, feeding and co-feeding) can potentially occur also in insects, particularly in mosquitoes. In the present study, adult as well as larval mosquitoes were collected at 42 different geographical locations throughout Germany. This is the first study, in which German mosquitoes were analyzed for the presence of Borrelia spp. Targeting two specific borrelial genes, flaB and ospA encoding for the subunit B of flagellin and the outer surface protein A, the results show that DNA of Borrelia afzelii, Borrelia bavariensis and Borrelia garinii could be detected in ten Culicidae species comprising four distinct genera (Aedes, Culiseta, Culex, and Ochlerotatus). Positive samples also include adult specimens raised in the laboratory from wild-caught larvae indicating that transstadial and/or transovarial transmission might occur within a given mosquito population.

  19. Amino acid sequence heterogeneity of the chromosomal encoded Borrelia burgdorferi sensu lato major antigen P100.

    Science.gov (United States)

    Fellinger, W; Farencena, A; Redl, B; Sambri, V; Cevenini, R; Stöffler, G

    1995-04-01

    The entire nucleotide sequence of the chromosomal encoded major antigen p100 of the European Borrelia garinii isolate B29 was determined and the deduced amino acid sequence was compared to the homologous antigen p83 of the North American Borrelia burgdorferi sensu stricto strain B31 and the p100 of the European Borrelia afzelii (group VS461) strain PKo. p100 of strain B29 shows 87% amino acid sequence identity to strain B31 and 79.2% to strain PKo, p100 of strain B31 and PKo shows 62.5% identity to each other. In addition, partial nucleotide sequences of the most heterogeneous region of the p100 gene of two other Borrelia garinii isolates (PBi and VS286) have been determined and the deduced amino acid sequences were compared with all p100 of Borrelia garinii published so far. We found an amino acid sequence identity between 88.6 and 100% within the same genospecies. The N-terminal part of the p100 proteins is highly conserved whereas a striking heterogeneous region within the C-terminal part of the proteins was observed.

  20. 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.

  1. 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

  2. Prevalence of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) in Ixodes scapularis (Acari: Ixodidae) adults in New Jersey, 2000-2001.

    Science.gov (United States)

    Schulze, Terry L; Jordan, Robert A; Hung, Robert W; Puelle, Rose S; Markowski, Daniel; Chomsky, Martin S

    2003-07-01

    Using polymerase chain reaction, we analyzed 529 Ixodes scapularis Say adults collected from 16 of New Jersey's 21 counties for the presence of Borrelia burgdorferi, the etiological agent of Lyme disease. Overall, 261 (49.3%) were positive. B. burgdorferi was detected in ticks obtained from each county and from 53 of the 58 (93.1%) municipalities surveyed. The observed statewide prevalence in New Jersey is similar to those reported from other northeastern and mid-Atlantic states. PMID:14680126

  3. 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...

  4. 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...

  5. Prevalence of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae) in Ixodes scapularis (Acari: Ixodidae) adults in New Jersey, 2000-2001.

    Science.gov (United States)

    Schulze, Terry L; Jordan, Robert A; Hung, Robert W; Puelle, Rose S; Markowski, Daniel; Chomsky, Martin S

    2003-07-01

    Using polymerase chain reaction, we analyzed 529 Ixodes scapularis Say adults collected from 16 of New Jersey's 21 counties for the presence of Borrelia burgdorferi, the etiological agent of Lyme disease. Overall, 261 (49.3%) were positive. B. burgdorferi was detected in ticks obtained from each county and from 53 of the 58 (93.1%) municipalities surveyed. The observed statewide prevalence in New Jersey is similar to those reported from other northeastern and mid-Atlantic states.

  6. 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...

  7. Role of Fc Gamma Receptors in Triggering Host Cell Activation and Cytokine Release by Borrelia burgdorferi

    Science.gov (United States)

    Talkington, Jeffrey; Nickell, Steven P.

    2001-01-01

    Borrelia burgdorferi, the spirochetal bacterium that causes human Lyme disease, encodes numerous lipoproteins which have the capacity to trigger the release of proinflammatory cytokines from a variety of host cell types, and it is generally believed that these cytokines contribute to the disease process in vivo. We previously reported that low-passage-number infectious B. burgdorferi spirochetes express a novel lipidation-independent activity which induces secretion of the proinflammatory cytokine tumor necrosis factor alpha (TNF-α) by the mouse MC/9 mast cell line. Using RNase protection assays, we determined that mast cells exposed in vitro to low-passage-number, but not high-passage-number, B. burgdorferi spirochetes show increased expression of additional mRNAs representing several chemokines, including macrophage-inflammatory protein 1α (MIP-1α), MIP-1β, and TCA3, as well as the proinflammatory cytokine interleukin-6. Furthermore, mast cell TNF-α secretion can be inhibited by the phosphatidylinositol 3-kinase inhibitor wortmannin and also by preincubation with purified mouse immunoglobulin G1 (IgG1) and IgG2a, but not mouse IgG3, and by a mouse Fc gamma receptor II and III (FcγRII/III)-specific rat monoclonal antibody, suggesting the likely involvement of host FcγRIII in B. burgdorferi-mediated signaling. A role for passively adsorbed rabbit or bovine IgG or serum components in B. burgdorferi-mediated FcγR signaling was excluded in control experiments. These studies confirm that low-passage-number B. burgdorferi spirochetes express a novel activity which upregulates the expression of a variety of host cell chemokine and cytokine genes, and they also establish a novel antibody-independent role for FcγRs in transduction of activation signals by bacterial products. PMID:11119532

  8. 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.

  9. 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.

  10. 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

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

    Directory of Open Access Journals (Sweden)

    Shakirat A. Adetunji

    2016-08-01

    Full Text Available 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.

  12. 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

  13. 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...

  14. 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.

  15. 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...

  16. 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...

  17. Linear chromosomal physical and genetic map of Borrelia burgdorferi, the Lyme disease agent.

    Science.gov (United States)

    Casjens, S; Huang, W M

    1993-05-01

    A physical map of the 952 kbp chromosome of Borrelia burgdorferi Sh-2-82 has been constructed. Eighty-three intervals on the chromosome, defined by the cleavage sites of 15 restriction enzymes, are delineated. The intervals vary in size from 96 kbp to a few hundred bp, with an average size of 11.5 kbp. A striking feature of the map is its linearity; no other bacterial groups are known to have linear chromosomes. The two ends of the chromosome do not hybridize with one another, indicating that there are no large common terminal regions. The chromosome of this strain was found to be stable in culture; passage 6, 165 and 320 cultures have identical chromosomal restriction maps. We have positioned all previously known Borrelia burgdorferi chromosomal genes and several newly identified ones on this map. These include the gyrA/gyrB/dnaA/dnaN gene cluster, the rRNA gene cluster, fla, flgE, groEL (hsp60), recA, the rho/hip cluster, the dnaK (hsp70)/dnaJ/grpE cluster, the pheT/pheS cluster, and the genes which encode the potent immunogen proteins p22A, p39 and p83. Our electrophoretic analysis detects five linear and at least two circular plasmids in B. burgdorferi Sh-2-82. We have constructed a physical map of the 53 kbp linear plasmid and located the operon that encodes the two major outer surface proteins ospA and ospB on this plasmid. Because of the absence of functional genetic tools for this organism, these maps will serve as a basis for future mapping, cloning and sequencing studies of B. burgdorferi.

  18. Risk of occupational infections caused by Borrelia burgdorferi among forestry workers and farmers

    Directory of Open Access Journals (Sweden)

    Małgorzata Tokarska-Rodak

    2014-02-01

    Full Text Available Background: The aim of the work was to analyze the incidence of infection with Borrelia burgdorferi in forestry workers and farmers, major groups occupationally exposed to tick bites. Material and Methods: The study group included 275 workers (171 foresters and 104 farmers. The control group consisted of 45 people, who have not been occupationally exposed to tick bites. The screening Elisa and Wb tests for the presence of anti-Borrelia IgM/IgG antibodies were performed in all subjects of the study and control groups. Statistical analysis was performed using the Chi2 test. Results: The positive results denoting the presence of anti-Borrelia IgM/IgG antibodies were found in 55% of farmers and 28% of foresters occupationally exposed to Lyme borreliosis and coming from the area of South Podlasie Lowland and Lublin Polesie. The differences between the forestry workers and the control group (p ≤ 0.00001 and between farmers and the control group (p ≤ 0.001 were statistically significant. The species, such as B. spielmanii and B. bavariensis, which have not yet been reported in Poland, are significant etiologic agents of Lyme disease. Conclusion: The risk of occupational exposure to the B. burgdorferi infection is high for foresters and farmers, and the infection with spirochetes is frequently confirmed on the basis of positive results of the Wb test. The presence of specific antibodies against protein antigens of B. spielmanii and B. bavariensis suggest that these bacteria can cause Lyme disease both independently and in participation with other Borrelia species, which influences the development of the clinical manifestations of infection. Med Pr 2014;65(1:109–117

  19. 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

  20. 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 ...

  1. 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.

  2. Human antibody responses to VlsE antigenic variation protein of Borrelia burgdorferi.

    Science.gov (United States)

    Lawrenz, M B; Hardham, J M; Owens, R T; Nowakowski, J; Steere, A C; Wormser, G P; Norris, S J

    1999-12-01

    VlsE is a 35-kDa surface-exposed lipoprotein of Borrelia burgdorferi that was shown previously to undergo antigenic variation through segmental recombination of silent vls cassettes with vlsE during experimental mouse infections. Previous data had indicated that sera from North American Lyme disease patients and experimentally infected animals contained antibodies reactive with VlsE. In this study, sera from patients with Lyme disease, syphilis, and autoimmune conditions as well as from healthy controls were examined for reactivity with VlsE by Western blotting and enzyme-linked immunosorbent assay (ELISA). Strong Western blot reactivity to a recombinant VlsE cassette region protein was obtained consistently with Lyme disease sera. Although sera from Lyme disease patients also reacted with a band corresponding to VlsE in B. burgdorferi B31-5A3, interpretation was complicated by low levels of VlsE expression in in vitro-cultured B. burgdorferi and by the presence of comigrating bands. An ELISA using recombinant VlsE was compared with an ELISA using sonically disrupted B. burgdorferi as the antigen. For a total of 93 Lyme disease patient sera examined, the VlsE ELISA yielded sensitivities of 63% for culture-confirmed erythema migrans cases and 92% for later stages, as compared to 61 and 98%, respectively, for the "whole-cell" ELISA. The specificities of the two assays with healthy blood donor sera were comparable, but the VlsE ELISA was 90% specific with sera from syphilis patients, compared to 20% specificity for the whole-cell ELISA with this group. Neither assay showed reactivity with a panel of sera from 20 non-Lyme disease arthritis patients or 20 systemic lupus erythematosus patients. Our results indicate that VlsE may be useful in the immunodiagnosis of Lyme disease and may offer greater specificity than ELISAs using whole B. burgdorferi as the antigen.

  3. 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

  4. 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...

  5. 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

  6. 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

  7. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

  9. 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.

  10. 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

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

    Directory of Open Access Journals (Sweden)

    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.

  12. 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.

  13. 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

  14. Borrelia burgdorferi genetic markers and disseminated disease in patients with early Lyme disease.

    Science.gov (United States)

    Jones, Kathryn L; Glickstein, Lisa J; Damle, Nitin; Sikand, Vijay K; McHugh, Gail; Steere, Allen C

    2006-12-01

    Three genetic markers of Borrelia burgdorferi have been associated with disseminated disease: the OspC type, the 16S-23S rRNA intergenic spacer type (RST), and vlsE. Here, we modified previous methods so as to identify the three markers by PCR and restriction fragment length polymorphism in parallel, analyzed B. burgdorferi isolates from erythema migrans (EM) skin lesions in 91 patients, and correlated the results with evidence of dissemination. OspC type A was found approximately twice as frequently in patients with disseminated disease, whereas type K was identified approximately twice as often in those without evidence of dissemination, but these trends were not statistically significant. The remaining seven types identified were found nearly equally in patients with or without evidence of dissemination. RST 1 strains were significantly associated with dissemination (P=0.03), whereas RST 2 and RST 3 strains tended to have an inverse association with this outcome. The vlsE gene was identified in all 91 cases, using primer sets specific for an N-terminal sequence of B. burgdorferi strain B31 (vlsEB31) or strain 297 (vlsE297), but neither marker was associated with dissemination. Specific combinations of the three genetic markers usually occurred together. OspC type A was always found with RST 1 and vlsEB31, type K was always identified with RST 2 and more often with vlsE297, and types E and I were almost always found with RST 3 and equally often with vlsEB31 and vlsE297. We conclude that B. burgdorferi strains vary in their capacity to disseminate, but almost all strains isolated from EM lesions sometimes caused disseminated disease.

  15. Borrelia burgdorferi Genetic Markers and Disseminated Disease in Patients with Early Lyme Disease▿

    Science.gov (United States)

    Jones, Kathryn L.; Glickstein, Lisa J.; Damle, Nitin; Sikand, Vijay K.; McHugh, Gail; Steere, Allen C.

    2006-01-01

    Three genetic markers of Borrelia burgdorferi have been associated with disseminated disease: the OspC type, the 16S-23S rRNA intergenic spacer type (RST), and vlsE. Here, we modified previous methods so as to identify the three markers by PCR and restriction fragment length polymorphism in parallel, analyzed B. burgdorferi isolates from erythema migrans (EM) skin lesions in 91 patients, and correlated the results with evidence of dissemination. OspC type A was found approximately twice as frequently in patients with disseminated disease, whereas type K was identified approximately twice as often in those without evidence of dissemination, but these trends were not statistically significant. The remaining seven types identified were found nearly equally in patients with or without evidence of dissemination. RST 1 strains were significantly associated with dissemination (P = 0.03), whereas RST 2 and RST 3 strains tended to have an inverse association with this outcome. The vlsE gene was identified in all 91 cases, using primer sets specific for an N-terminal sequence of B. burgdorferi strain B31 (vlsEB31) or strain 297 (vlsE297), but neither marker was associated with dissemination. Specific combinations of the three genetic markers usually occurred together. OspC type A was always found with RST 1 and vlsEB31, type K was always identified with RST 2 and more often with vlsE297, and types E and I were almost always found with RST 3 and equally often with vlsEB31 and vlsE297. We conclude that B. burgdorferi strains vary in their capacity to disseminate, but almost all strains isolated from EM lesions sometimes caused disseminated disease. PMID:17035489

  16. Diversity of antibody responses to Borrelia burgdorferi in experimentally infected beagle dogs.

    Science.gov (United States)

    Baum, Elisabeth; Grosenbaugh, Deborah A; Barbour, Alan G

    2014-06-01

    Lyme borreliosis (LB) is a common infection of domestic dogs in areas where there is enzootic transmission of the agent Borrelia burgdorferi. While immunoassays based on individual subunits have mostly supplanted the use of whole-cell preparations for canine serology, only a limited number of informative antigens have been identified. To more broadly characterize the antibody responses to B. burgdorferi infection and to assess the diversity of those responses in individual dogs, we examined sera from 32 adult colony-bred beagle dogs that had been experimentally infected with B. burgdorferi through tick bites and compared those sera in a protein microarray with sera from uninfected dogs in their antibody reactivities to various recombinant chromosome- and plasmid-encoded B. burgdorferi proteins, including 24 serotype-defining OspC proteins of North America. The profiles of immunogenic proteins for the dogs were largely similar to those for humans and natural-reservoir rodents; these proteins included the decorin-binding protein DbpB, BBA36, BBA57, BBA64, the fibronectin-binding protein BBK32, VlsE, FlaB and other flagellar structural proteins, Erp proteins, Bdr proteins, and all of the OspC proteins. In addition, the canine sera bound to the presumptive lipoproteins BBB14 and BB0844, which infrequently elicited antibodies in humans or rodents. Although the beagle, like most other domestic dog breeds, has a small effective population size and features extensive linkage disequilibrium, the group of animals studied here demonstrated diversity in antibody responses in measures of antibody levels and specificities for conserved proteins, such as DbpB, and polymorphic proteins, such as OspC.

  17. 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

  18. 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.

  19. 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.

  20. 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

  1. 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...

  2. 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...

  3. 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

  4. Development of a Multiantigen Panel for Improved Detection of Borrelia burgdorferi Infection in Early Lyme Disease.

    Science.gov (United States)

    Lahey, Lauren J; Panas, Michael W; Mao, Rong; Delanoy, Michelle; Flanagan, John J; Binder, Steven R; Rebman, Alison W; Montoya, Jose G; Soloski, Mark J; Steere, Allen C; Dattwyler, Raymond J; Arnaboldi, Paul M; Aucott, John N; Robinson, William H

    2015-12-01

    The current standard for laboratory diagnosis of Lyme disease in the United States is serologic detection of antibodies against Borrelia burgdorferi. The Centers for Disease Control and Prevention recommends a two-tiered testing algorithm; however, this scheme has limited sensitivity for detecting early Lyme disease. Thus, there is a need to improve diagnostics for Lyme disease at the early stage, when antibiotic treatment is highly efficacious. We examined novel and established antigen markers to develop a multiplex panel that identifies early infection using the combined sensitivity of multiple markers while simultaneously maintaining high specificity by requiring positive results for two markers to designate a positive test. Ten markers were selected from our initial analysis of 62 B. burgdorferi surface proteins and synthetic peptides by assessing binding of IgG and IgM to each in a training set of Lyme disease patient samples and controls. In a validation set, this 10-antigen panel identified a higher proportion of early-Lyme-disease patients as positive at the baseline or posttreatment visit than two-tiered testing (87.5% and 67.5%, respectively; P Lyme disease.

  5. Borrelia burgdorferi infection regulates CD1 expression in human cells and tissues via IL1-β.

    Science.gov (United States)

    Yakimchuk, Konstantin; Roura-Mir, Carme; Magalhaes, Kelly G; de Jong, Annemieke; Kasmar, Anne G; Granter, Scott R; Budd, Ralph; Steere, Allen; Pena-Cruz, Victor; Kirschning, Carsten; Cheng, Tan-Yun; Moody, D Branch

    2011-03-01

    The appearance of group 1 CD1 proteins (CD1a, CD1b and CD1c) on maturing myeloid DC is a key event that converts myeloid DC to effective lipid APC. Here, we show that Borrelia burgdorferi, the causative agent of Lyme disease, triggers appearance of group 1 CD1 proteins at high density on the surface of human myeloid DC during infection. Within human skin, CD1b and CD1c expression was low or absent prior to infection, but increased significantly after experimental infections and in erythema migrans lesions from Lyme disease patients. The induction of CD1 was initiated by borrelial lipids acting through TLR-2 within minutes, but required 3 days for maximum effect. The delay in CD1 protein appearance involved a multi-step process whereby TLR-2 stimulated cells release soluble factors, which are sufficient to transfer the CD1-inducing effect in trans to other cells. Analysis of these soluble factors identified IL-1β as a previously unknown pathway leading to group 1 CD1 protein function. This study establishes that upregulation of group 1 CD1 proteins is an early event in B. burgdorferi infection and suggests a stepwise mechanism whereby bacterial cell walls, TLR activation and cytokine release cause DC precursors to express group 1 CD1 proteins.

  6. 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

  7. 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

  8. 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

  9. 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

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

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    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.

  11. Mechanisms of infection by pathogens transmitted by ticks on the example of bacteria: Anaplasma phagocytophilum and Borrelia burgdorferi

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    Paula Wróblewska

    2016-06-01

    Full Text Available Tick-borne diseases are transmission diseases belonging to the group of zoonoses but carried by ticks. These diseases are a major public health problem but also a problem for groups occupationally exposed to tick bites. Ixodes ricinus is a species of ticks which is the most common reservoir and the vector of a large number of microorganisms pathogenic to humans. It transfers, among others, bacteria of the species: Anaplasma phagocytophilum and Borrelia burgdorferi. The article discusses the mechanisms of infection with Borrelia burgdorferi and Anaplasma phagocytophilum for both ticks as well as for animals and humans. The two microorganisms discussed have developed many characteristics and mechanisms of adaptation to the environment, as well as defense mechanisms against the body's immune response. Understanding the biology of ticks and the function of proteins produced by ticks and pathogenic microorganisms is the key in the development of effective treatments and prevention of Lyme disease and anaplasmosis.

  12. Selective association of outer surface lipoproteins with the lipid rafts of Borrelia burgdorferi.

    Science.gov (United States)

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

    2014-03-11

    Borrelia burgdorferi contains unique cholesterol-glycolipid-rich lipid rafts that are associated with lipoproteins. These complexes suggest the existence of macromolecular structures that have not been reported for prokaryotes. Outer surface lipoproteins OspA, OspB, and OspC were studied for their participation in the formation of lipid rafts. Single-gene deletion mutants with deletions of ospA, ospB, and ospC and a spontaneous gene mutant, strain B313, which does not express OspA and OspB, were used to establish their structural roles in the lipid rafts. All mutant strains used in this study produced detergent-resistant membranes, a common characteristic of lipid rafts, and had similar lipid and protein slot blot profiles. Lipoproteins OspA and OspB but not OspC were shown to be associated with lipid rafts by transmission electron microscopy. When the ability to form lipid rafts in live B. burgdorferi spirochetes was measured by fluorescence resonance energy transfer (FRET), strain B313 showed a statistically significant lower level of segregation into ordered and disordered membrane domains than did the wild-type and the other single-deletion mutants. The transformation of a B313 strain with a shuttle plasmid containing ospA restored the phenotype shared by the wild type and the single-deletion mutants, demonstrating that OspA and OspB have redundant functions. In contrast, a transformed B313 overexpressing OspC neither rescued the FRET nor colocalized with the lipid rafts. Because these lipoproteins are expressed at different stages of the life cycle of B. burgdorferi, their selective association is likely to have an important role in the structure of prokaryotic lipid rafts and in the organism's adaptation to changing environments. IMPORTANCE Lipid rafts are cholesterol-rich clusters within the membranes of cells. Lipid rafts contain proteins that have functions in sensing the cell environment and transmitting signals. Although selective proteins are present in

  13. 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...

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

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    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...

  15. 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...

  16. 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

  17. First detection of Borrelia burgdorferi-antibodies in free-living birds of prey from Eastern Westphalia, Germany.

    Science.gov (United States)

    Büker, M; Picozzi, K; Kolb, S; Hatt, J-M

    2013-07-01

    Borrelia (B.) burgdorferi sensu lato, the causative agent of Lyme disease, is the most important arthropod-borne zoonosis-pathogen in the Northern hemisphere. Besides small mammals, birds, primarily Passeriformes and sea birds, play an important role in the transmission, distribution and maintenance of this disease. Previous studies on birds have focused mainly on the detection of Borrelia-infected ticks. However, the presence or absence of an infected tick cannot be taken as an indicator of the infective status of the avian host; to date this area of research has not been explored. In this study, serological analyses of blood collected from free-living birds of prey (n = 29) at the rehabilitation centre in Eastern Westphalia, Germany, highlights that birds of prey are also susceptible to B. burgdorferi and react immunologically to an infection. Increased antibody-levels could be found by using a modified Indirect Immunofluorescent-testing in two common buzzards, Buteo buteo, and two eagle owls, Bubo bubo. Further research regarding the serological diagnostics of B. burgdorferi within the avian host is required. In the future, it should be taken into account that birds of prey can be reservoirs for B. burgdorferi, as well as carriers of infected ticks; although at present their epidemiological importance is still to be confirmed. PMID:23823746

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

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    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.

  19. Borrelia burgdorferi Induces TLR2-Mediated Migration of Activated Dendritic Cells in an Ex Vivo Human Skin Model

    Science.gov (United States)

    Wagemakers, Alex; van ‘t Veer, Cornelis; Oei, Anneke; van der Pot, Wouter J.; Ahmed, Kalam; van der Poll, Tom; Geijtenbeek, Teunis B. H.; Hovius, Joppe W. R.

    2016-01-01

    Borrelia burgdorferi is transmitted into the skin of the host where it encounters and interacts with two dendritic cell (DC) subsets; Langerhans cells (LCs) and dermal DCs (DDCs). These cells recognize pathogens via pattern recognition receptors, mature and migrate out of the skin into draining lymph nodes, where they orchestrate adaptive immune responses. In order to investigate the response of skin DCs during the early immunopathogenesis of Lyme borreliosis, we injected B. burgdorferi intradermally into full-thickness human skin and studied the migration of DCs out of the skin, the activation profile and phenotype of migrated cells. We found a significant increase in the migration of LCs and DDCs in response to B. burgdorferi. Notably, migration was prevented by blocking TLR2. DCs migrated from skin inoculated with higher numbers of spirochetes expressed significantly higher levels of CD83 and produced pro-inflammatory cytokines. No difference was observed in the expression of HLA-DR, CD86, CD38, or CCR7. To conclude, we have established an ex vivo human skin model to study DC-B. burgdorferi interactions. Using this model, we have demonstrated that B. burgdorferi-induced DC migration is mediated by TLR2. Our findings underscore the utility of this model as a valuable tool to study immunity to spirochetal infections. PMID:27695100

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

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    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.

  1. Interpretation criteria for standardized Western blots for three European species of Borrelia burgdorferi sensu lato.

    Science.gov (United States)

    Hauser, U; Lehnert, G; Lobentanzer, R; Wilske, B

    1997-06-01

    Western blots (WBs; immunoblots) are a widely used tool for the serodiagnosis of Lyme borreliosis, but so far, no defined criteria for performance, analysis, and interpretation have been established in Europe. For the current study WBs were produced with strains PKa2 (Borrelia burgdorferi sensu stricto), PKo (Borrelia afzelii), and PBi (Borrelia garinii). To improve resolution we used gels of 17 cm in length. In a first step, 13 immunodominant proteins were identified with monoclonal antibodies. Then, the apparent molecular masses of all visually distinguishable bands were determined densitometrically. Approximately 40 bands of between 14 and 100 kDa were differentiated for each strain. From a study with 330 serum samples (from 189 patients with Lyme borreliosis and 141 controls), all observed bands were documented. To establish criteria for a positive WB result, the discriminating ability of a series of band combinations (interpretation rules) were evaluated separately for each strain (for immunoglobulin G [IgG] WB, > 40 combinations; for IgM WB, > 15 combinations). The following interpretation criteria resulting in specificities of greater than 96% were recommended: for IgG WB, at least one band of p83/100, p58, p56, OspC, p21, and p17a for PKa2; at least two bands of p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo; and at least one band of p83/100, p39, OspC, p21, and p17b for PBi; for IgM WB, at least one band of p39, OspC, and p17a or a strong p41 band for PKa2; at least one band of p39, OspC, and p17 or a strong p41 band for PKo; and at least one band of p39 and OspC or a strong p41 band for PBi. The overall sensitivity was the highest for PKo WB, followed by PBi and PKa2 WB, in decreasing order. Standardization of WB assays is necessary for comparison of results from different laboratories.

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

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    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

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

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    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

  4. 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.

  5. 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

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

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    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.

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

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    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.

  8. 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...

  9. 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...

  10. Synthesis of Th17 cytokines in the culture of peripheral blood mononuclear cells stimulated with Borrelia burgdorferi sensu lato

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    Sambor Grygorczuk

    2016-06-01

    Full Text Available [b]Introduction and objective. [/b]Th17 lymphocytes and their cytokines, interleukin 17A (IL-17A, IL-17F and IL-22, participate in the response to extracellular bacteria and in the autoimmunity and may be engaged in the pathogenesis of Lyme borreliosis. Concentrations were measured of IL-17A, IL-17F and IL-22 in the supernatant of the peripheral blood mononuclear cells (PBMC culture stimulated with [i]Borrelia burgdorferi sensu lato[/i] ([i]B. burgdorferi[/i]. [b]Materials and method.[/b] The study group consisted of 13 patients with early disseminated and late Lyme borreliosis and a control group of 7 healthy persons. PBMC cultures were stimulated for 48 hours with [i]B. burgdorferi [/i]spirochetes of three pathogenic species: [i]B. burgdorferi[/i] sensu stricto, B. afzelii or B. garinii, in the multiplicity of infection 10:1. Concentrations of Th17 cytokines IL-17A, IL-17F and IL-22, as well as Th2/immunoregulatory cytokine IL-10 were measured with ELISA assays. [b]Results. [/b]Expression of IL-17A, IL-17F and IL-22 increased under stimulation, simultaneously with the increased IL-10 expression. Concentration of IL-17F tended to be lower in early neuroborreliosis than in late Lyme borreliosis and than in controls. [i]B. afzelii[/i] elicited higher expression of IL-17A than the other two species. [b]Conclusions.[/b] IL-17A, IL-17F and IL-22 are synthesized simultaneously by PBMC stimulated with [i]B. burgdorferi[/i]. There is no antagonism between Th17 response and IL-10 expression. The role of Th17 cytokines seems to differ depending on the clinical stage of Lyme borreliosis and on the [i]B. burgdorferi[/i] species.

  11. 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.

  12. 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.

  13. 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 ...

  14. 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

  15. Persistence of immunoglobulin M or immunoglobulin G antibody responses to Borrelia burgdorferi 10-20 years after active Lyme disease.

    Science.gov (United States)

    Kalish, R A; McHugh, G; Granquist, J; Shea, B; Ruthazer, R; Steere, A C

    2001-09-15

    The interpretation of serological results for patients who had Lyme disease many years ago is not well defined. We studied the serological status of 79 patients who had had Lyme disease 10-20 years ago and did not currently have signs or symptoms of active Lyme disease. Of the 40 patients who had had early Lyme disease alone, 4 (10%) currently had IgM responses to Borrelia burgdorferi, and 10 (25%) still had IgG reactivity to the spirochete, as determined by a 2-test approach (enzyme-linked immunosorbent assay and Western blot). Of the 39 patients who had had Lyme arthritis, 6 (15%) currently had IgM responses and 24 (62%) still had IgG reactivity to the spirochete. IgM or IgG antibody responses to B. burgdorferi may persist for 10-20 years, but these responses are not indicative of active infection.

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

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    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.

  17. 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.

  18. 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...

  19. 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

  20. Molecular and immunological characterization of the p83/100 protein of various Borrelia burgdorferi sensu lato strains.

    Science.gov (United States)

    Rössler, D; Eiffert, H; Jauris-Heipke, S; Lehnert, G; Preac-Mursic, V; Teepe, J; Schlott, T; Soutschek, E; Wilske, B

    1995-05-01

    The complete coding regions of the chromosomally encoded p83/100 protein of four Borrelia garinii strains and one Borrelia burgdorferi sensu stricto strain have been amplified by the polymerase chain reaction (PCR), cloned and sequenced. From alignment studies with the deduced amino acid sequences presented here, and five other published p83/100 sequences, the most heterologous region of the p83/100 molecule was identified to be located between amino acid position 390-540. To study the structure of this heterogeneous region, and internal fragment of the p83/100 genes from 11 additional B. burgdorferi sensu lato strains was amplified by PCR. The PCR products were analyzed by DNA sequencing and restriction enzyme analysis. These internal p83/100 fragments varied in size and sequence. Cluster analysis of internal p83/100 fragments, as well as restriction enzyme analysis, revealed three major groups in accordance with grouping into the three species causing Lyme disease. Strains within the same species (six B. burgdorferi sensu stricto and six B. afzelii strains) showed similar p83/100 partial structures. Nevertheless, nine B. garinii strains showed more sequence variations and could be further divided into two major subgroups. One group is represented by OspA serotype 4 strains, the other more heterogeneous group is represented by OspA serotypes 3, 5, 6 and 7 strains. Phenotypic analysis with four p83/100-specific monoclonal antibodies revealed four distinct reactivity patterns. Antibody L100 1B4 recognized a common epitope of B. burgdorferi sensu stricto and B. afzelii. Antibodies L100 17D3 and L100 18B4 were reactive with an epitope shared by strains of all three species. The broadest reactivity was shown by L100 18B4 which, in contrast to L100 17D3, additionally recognized the relapsing fever borreliae B. turicatae and B. hermsii. L100 8B8 detected a subgroup of the B. burgdorferi sensu stricto strains. Since comparison of the p83/100 molecule with sequences from

  1. 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...

  2. 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.

  3. 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. PMID:27504018

  4. Borrelia burgdorferi genospecies detection by RLB hybridization in Ixodes cinus ticks from different sites of North-Eastern Poland

    Directory of Open Access Journals (Sweden)

    Justyna Dunaj

    2014-06-01

    Full Text Available Introduction. RLB (Reverse Line Blot Hybridization is a molecular biology technique that might be used for [i]Borrelia burgdorferi [/i]sensu lato (sl DNA detection with genospecies specification. Among[i] B. burgdorferi[/i] sl genospecies at least 7 are regarded as pathogenic in Europe. objective. The aim of the study was to evaluate the frequency of different [i]Borrelia[/i] genospecies DNA detection in Ixodes ricinus ticks in the endemic area of North-Eastern Poland by using RLB. materials and method. Ixodes ricinus ticks were collected in May – June, from 6 different sites in North-Eastern Poland (Jakubin, Kolno, Grajewo, Suwałki, Siemiatycze, Białowieża by flagging. Extracted DNA was amplified by polymerase chain reaction (PCR targeting the intergenic spacer 5S 23S of [i]B. burgdorferi sl.[/i] PCR products were hybridised to 15 different oligonucleotide probes for 9 different [i]Borrelia [/i]genospecies ([i]B. burgdorferi sl, B. burgdorferi ss, B. garinii, B. afzelii, B. valaisiana, B. lusitaniae, B. spielmanii, B. bissettii and B. relapsing[/i] fever-like spirochetes (B. myamotoi by RLB. results. [i]Borrelia [/i]genospecies DNA was detected in 205 Ixodes ricinus ticks. Among 14 infected with [i]Borrelia[/i] ticks, 4 were identified as B. garinii and 10 as B. afzelii. Higher numbers of infected ticks were noticed in the eastern part of the research area, where large forest complexes dominate. Nymphs appeared to be the most frequently infected tick stage, which has an epidemiological meaning in the incidence of Lyme borreliosis. conclusions. The study demonstrated that RLB might be easily used in [i]Borrelia[/i] DNA detection with genospecies-identification, and indicated the domination of [i]B. afzelii and B. garinii [/i]in ticks from North-Eastern Poland.

  5. Live Borrelia burgdorferi preferentially activate interleukin-1 beta gene expression and protein synthesis over the interleukin-1 receptor antagonist.

    Science.gov (United States)

    Miller, L C; Isa, S; Vannier, E; Georgilis, K; Steere, A C; Dinarello, C A

    1992-01-01

    Lyme arthritis is one of the few forms of chronic arthritis in which the cause is known with certainty. Because cytokines are thought to contribute to the pathogenesis of chronic arthritis, we investigated the effect of the Lyme disease spirochete, Borrelia burgdorferi, on the gene expression and synthesis of IL-1 beta and the IL-1 receptor antagonist (IL-1ra) in human peripheral blood mononuclear cells. Live B. burgdorferi induced fivefold more IL-1 beta than IL-1 alpha and sevenfold more IL-1 beta than IL-1ra; LPS or sonicated B. burgdorferi induced similar amounts of all three cytokines. This preferential induction of IL-1 beta was most dramatic in response to a low passage, virulent preparation of B. burgdorferi vs. three high passage avirulent strains. No difference in induction of IL-1ra was seen between these strains. The marked induction of IL-1 beta was partially diminished by heat-treatment and abrogated by sonication; IL-1ra was not affected. This suggested that a membrane component(s) accounted for the preferential induction of IL-1 beta. However, recombinant outer surface protein beta induced little IL-1 beta. By 4 h after stimulation, B. burgdorferi induced sixfold more IL-1 beta protein than LPS. In contrast to LPS-induced IL-1 beta mRNA which reached maximal accumulation after 3 h, B. burgdorferi-induced IL-1 beta mRNA showed biphasic elevations at 3 and 18 h. B. burgdorferi-induced IL-1ra mRNA peaked at 12 h, whereas LPS-induced IL-1ra mRNA peaked at 9 h. IL-1 beta synthesis increased in response to increasing numbers of spirochetes, whereas IL-1ra synthesis did not. The preferential induction by B. burgdorferi of IL-1 beta over IL-1ra is an example of excess agonist over antagonist synthesis induced by a microbial pathogen, and may contribute to the destructive lesion of Lyme arthritis. Images PMID:1387885

  6. 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

  7. 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...

  8. 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

  9. 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.

  10. Characterization of the RelBbu Regulon in Borrelia burgdorferi Reveals Modulation of Glycerol Metabolism by (p)ppGpp.

    Science.gov (United States)

    Bugrysheva, Julia V; Pappas, Christopher J; Terekhova, Darya A; Iyer, Radha; Godfrey, Henry P; Schwartz, Ira; Cabello, Felipe C

    2015-01-01

    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 (p)ppGpp) 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 (p)ppGpp 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.

  11. Borrelia burgdorferi DNA in the urine of treated patients with chronic Lyme disease symptoms. A PCR study of 97 cases.

    Science.gov (United States)

    Bayer, M E; Zhang, L; Bayer, M H

    1996-01-01

    The presence of Borrelia burgdorferi DNA was established by PCR from urine samples of 97 patients clinically diagnosed as presenting with symptoms of chronic Lyme disease. All patients had shown erythema chronica migrans following a deer tick bite. Most of the patients had been antibiotic-treated for extended periods of time. We used three sets of primer pairs with DNA sequences for the gene coding of outer surface protein A (OspA) and of a genomic sequence of B. burgdorferi to study samples of physician-referred patients from the mideastern USA. Controls from 62 healthy volunteers of the same geographic areas were routinely carried through the procedures in parallel with patients' samples. Of the 97 patients, 72 (74.2%) were found with positive PCR and the rest with negative PCR. The 62 healthy volunteers were PCR negative. It is proposed that a sizeable group of patients diagnosed on clinical grounds as having chronic Lyme disease may still excrete Borrelia DNA, and may do so in spite of intensive antibiotic treatment.

  12. 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

  13. Homogeneous Inflammatory Gene Profiles Induced in Human Dermal Fibroblasts in Response to the Three Main Species of Borrelia burgdorferi sensu lato

    Science.gov (United States)

    Meddeb, Mariam; Carpentier, Wassila; Cagnard, Nicolas; Nadaud, Sophie; Grillon, Antoine; Barthel, Cathy; De Martino, Sylvie Josiane; Jaulhac, Benoît; Boulanger, Nathalie

    2016-01-01

    In Lyme borreliosis, the skin is the key site for bacterial inoculation by the infected tick and for cutaneous manifestations. We previously showed that different strains of Borrelia burgdorferi sensu stricto isolated from tick and from different clinical stages of the Lyme borreliosis (erythema migrans, and acrodermatitis chronica atrophicans) elicited a very similar transcriptional response in normal human dermal fibroblasts. In this study, using whole transcriptome microarray chips, we aimed to compare the transcriptional response of normal human dermal fibroblasts stimulated by 3 Borrelia burgdorferi sensu lato strains belonging to 3 main pathogenic species (B. afzelii, B. garinii and B. burgdorferi sensu stricto) in order to determine whether “species-related” inflammatory pathways could be identified. The three Borrelia strains tested exhibited similar transcriptional profiles, and no species-specific fingerprint of transcriptional changes in fibroblasts was observed. Conversely, a common core of chemokines/cytokines (CCL2, CXCL1, CXCL2, CXCL6, CXCL10, IL-6, IL-8) and interferon-related genes was stimulated by all the 3 strains. Dermal fibroblasts appear to play a key role in the cutaneous infection with Borrelia, inducing a homogeneous inflammatory response, whichever Borrelia species was involved. PMID:27706261

  14. 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

  15. Establishment of enzyme-linked immunosorbent assay using purified recombinant 83-kilodalton antigen of Borrelia burgdorferi sensu stricto and Borrelia afzelii for serodiagnosis of Lyme disease.

    Science.gov (United States)

    Rauer, S; Kayser, M; Neubert, U; Rasiah, C; Vogt, A

    1995-10-01

    The 83-kDa antigen of Borrelia burgdorferi was expressed as a recombinant protein in Escherichia coli and purified for use in an enzyme-linked immunosorbent assay (p83-ELISA). Antibodies to the 83-kDa antigen of both the immunoglobulin G (IgG) and IgM isotypes could be detected in all stages of Lyme disease. Sensitivity varied, depending on the clinical stage of illness. In early stages, as defined for 118 patients with erythema migrans, it was found to be 20% (24 of 118 patients: 7 with IgM, 16 with IgG, and 1 with IgM and IgG). Of the patients with late-stage Lyme arthritis and acrodermatitis chronica atrophicans, 94% (16 of 17:2 with IgM and IgG and 14 with IgG) and 86% (36 of 42:2 with IgG and IgM and 34 with IgG) revealed positive results in the p83-ELISA, respectively. p83 displays sequence heterogeneity according to the genomospecies, but when the reactions of serum specimens from acrodermatitis chronica atrophicans patients and arthritis patients with p83 derived from representative strains of B. burgdorferi sensu stricto and Borrelia afzelii in ELISAs were compared, no differences in specificity and sensitivity were seen. When 82 serum specimens from healthy controls were tested, none had IgG and only 3 (4%) had IgM antibodies, indicating a high specificity. Positive reactions with antibodies against Treponema pallidum (1 of 37 patients; IgG) and Epstein-Barr virus (1 of 44 patients; IgM) and with autoantibodies of various specificities (1 of 53 patients; IgG) were seen with p83-ELISA provided little diagnostic information for Lyme disease, whereas the IgG p83-ELISA appears to be a suita ;e test for serodiagnosis of advanced-stage Lyme disease.

  16. Microarray analyses of inflammation response of human dermal fibroblasts to different strains of Borrelia burgdorferi sensu stricto.

    Directory of Open Access Journals (Sweden)

    Frédéric Schramm

    Full Text Available In Lyme borreliosis, the skin is the key site of bacterial inoculation by the infected tick, and of cutaneous manifestations, erythema migrans and acrodermatitis chronica atrophicans. We explored the role of fibroblasts, the resident cells of the dermis, in the development of the disease. Using microarray experiments, we compared the inflammation of fibroblasts induced by three strains of Borrelia burgdorferi sensu stricto isolated from different environments and stages of Lyme disease: N40 (tick, Pbre (erythema migrans and 1408 (acrodermatitis chronica atrophicans. The three strains exhibited a similar profile of inflammation with strong induction of chemokines (CXCL1 and IL-8 and IL-6 cytokine mainly involved in the chemoattraction of immune cells. Molecules such as TNF-alpha and NF-κB factors, metalloproteinases (MMP-1, -3 and -12 and superoxide dismutase (SOD2, also described in inflammatory and cellular events, were up-regulated. In addition, we showed that tick salivary gland extracts induce a cytotoxic effect on fibroblasts and that OspC, essential in the transmission of Borrelia to the vertebrate host, was not responsible for the secretion of inflammatory molecules by fibroblasts. Tick saliva components could facilitate the early transmission of the disease to the site of injury creating a feeding pit. Later in the development of the disease, Borrelia would intensively multiply in the skin and further disseminate to distant organs.

  17. A tick gut protein with fibronectin III domains aids Borrelia burgdorferi congregation to the gut during transmission.

    Science.gov (United States)

    Narasimhan, Sukanya; Coumou, Jeroen; Schuijt, Tim J; Boder, Eric; Hovius, Joppe W; Fikrig, Erol

    2014-08-01

    Borrelia burgdorferi transmission to the vertebrate host commences with growth of the spirochete in the tick gut and migration from the gut to the salivary glands. This complex process, involving intimate interactions of the spirochete with the gut epithelium, is pivotal to transmission. We utilized a yeast surface display library of tick gut proteins to perform a global screen for tick gut proteins that might interact with Borrelia membrane proteins. A putative fibronectin type III domain-containing tick gut protein (Ixofin3D) was most frequently identified from this screen and prioritized for further analysis. Immunization against Ixofin3D and RNA interference-mediated reduction in expression of Ixofin3D resulted in decreased spirochete burden in tick salivary glands and in the murine host. Microscopic examination showed decreased aggregation of spirochetes on the gut epithelium concomitant with reduced expression of Ixofin3D. Our observations suggest that the interaction between Borrelia and Ixofin3D facilitates spirochete congregation to the gut during transmission, and provides a "molecular exit" direction for spirochete egress from the gut.

  18. A tick gut protein with fibronectin III domains aids Borrelia burgdorferi congregation to the gut during transmission.

    Directory of Open Access Journals (Sweden)

    Sukanya Narasimhan

    2014-08-01

    Full Text Available Borrelia burgdorferi transmission to the vertebrate host commences with growth of the spirochete in the tick gut and migration from the gut to the salivary glands. This complex process, involving intimate interactions of the spirochete with the gut epithelium, is pivotal to transmission. We utilized a yeast surface display library of tick gut proteins to perform a global screen for tick gut proteins that might interact with Borrelia membrane proteins. A putative fibronectin type III domain-containing tick gut protein (Ixofin3D was most frequently identified from this screen and prioritized for further analysis. Immunization against Ixofin3D and RNA interference-mediated reduction in expression of Ixofin3D resulted in decreased spirochete burden in tick salivary glands and in the murine host. Microscopic examination showed decreased aggregation of spirochetes on the gut epithelium concomitant with reduced expression of Ixofin3D. Our observations suggest that the interaction between Borrelia and Ixofin3D facilitates spirochete congregation to the gut during transmission, and provides a "molecular exit" direction for spirochete egress from the gut.

  19. 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

  20. 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

  1. 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.

  2. 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

  3. In Vivo Imaging Demonstrates That Borrelia burgdorferi ospC Is Uniquely Expressed Temporally and Spatially throughout Experimental Infection

    Science.gov (United States)

    Skare, Jonathan T.; Shaw, Dana K.; Trzeciakowski, Jerome P.

    2016-01-01

    Borrelia burgdorferi is a spirochetal bacterium transmitted by the Ixodes tick that causes Lyme disease in humans due to its ability to evade the host immune response and disseminate to multiple immunoprotective tissues. The pathogen undergoes dynamic genetic alterations important for adaptation from the tick vector to the mammalian host, but little is known regarding the changes at the transcriptional level within the distal tissues they colonize. In this study, B. burgdorferi infection and gene expression of the essential virulence determinant ospC was quantitatively monitored in a spatial and temporal manner utilizing reporter bioluminescent borrelial strains with in vivo and ex vivo imaging. Although expressed from a shuttle vector, the PospC-luc construct exhibited a similar expression pattern relative to native ospC. Bacterial burden in skin, inguinal lymph node, heart, bladder and tibiotarsal joint varied between tissues and fluctuated over the course of infection possibly in response to unique cues of each microenvironment. Expression of ospC, when normalized for changes in bacterial load, presented unique profiles in murine tissues at different time points. The inguinal lymph node was infected with a significant B. burgdorferi burden, but showed minimal ospC expression. B. burgdorferi infected skin and heart induced expression of ospC early during infection while the bladder and tibiotarsal joint continued to display PospC driven luminescence throughout the 21 day time course. Localized skin borrelial burden increased dramatically in the first 96 hours following inoculation, which was not paralleled with an increase in ospC expression, despite the requirement of ospC for dermal colonization. Quantitation of bioluminescence representing ospC expression in individual tissues was validated by qRT-PCR of the native ospC transcript. Taken together, the temporal regulation of ospC expression in distal tissues suggests a role for this virulence determinant beyond

  4. Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking Activation of the C1 Complement Complex.

    Science.gov (United States)

    Garcia, Brandon L; Zhi, Hui; Wager, Beau; Höök, Magnus; Skare, Jon T

    2016-01-01

    Pathogens that traffic in blood, lymphatics, or interstitial fluids must adopt strategies to evade innate immune defenses, notably the complement system. Through recruitment of host regulators of complement to their surface, many pathogens are able to escape complement-mediated attack. The Lyme disease spirochete, Borrelia burgdorferi, produces a number of surface proteins that bind to factor H related molecules, which function as the dominant negative regulator of the alternative pathway of complement. Relatively less is known about how B. burgdorferi evades the classical pathway of complement despite the observation that some sensu lato strains are sensitive to classical pathway activation. Here we report that the borrelial lipoprotein BBK32 potently and specifically inhibits the classical pathway by binding with high affinity to the initiating C1 complex of complement. In addition, B. burgdorferi cells that produce BBK32 on their surface bind to both C1 and C1r and a serum sensitive derivative of B. burgdorferi is protected from killing via the classical pathway in a BBK32-dependent manner. Subsequent biochemical and biophysical approaches localized the anti-complement activity of BBK32 to its globular C-terminal domain. Mechanistic studies reveal that BBK32 acts by entrapping C1 in its zymogen form by binding and inhibiting the C1 subcomponent, C1r, which serves as the initiating serine protease of the classical pathway. To our knowledge this is the first report of a spirochetal protein acting as a direct inhibitor of the classical pathway and is the only example of a biomolecule capable of specifically and noncovalently inhibiting C1/C1r. By identifying a unique mode of complement evasion this study greatly enhances our understanding of how pathogens subvert and potentially manipulate host innate immune systems.

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

    Directory of Open Access Journals (Sweden)

    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.

  6. In Vivo Imaging Demonstrates That Borrelia burgdorferi ospC Is Uniquely Expressed Temporally and Spatially throughout Experimental Infection.

    Science.gov (United States)

    Skare, Jonathan T; Shaw, Dana K; Trzeciakowski, Jerome P; Hyde, Jenny A

    2016-01-01

    Borrelia burgdorferi is a spirochetal bacterium transmitted by the Ixodes tick that causes Lyme disease in humans due to its ability to evade the host immune response and disseminate to multiple immunoprotective tissues. The pathogen undergoes dynamic genetic alterations important for adaptation from the tick vector to the mammalian host, but little is known regarding the changes at the transcriptional level within the distal tissues they colonize. In this study, B. burgdorferi infection and gene expression of the essential virulence determinant ospC was quantitatively monitored in a spatial and temporal manner utilizing reporter bioluminescent borrelial strains with in vivo and ex vivo imaging. Although expressed from a shuttle vector, the PospC-luc construct exhibited a similar expression pattern relative to native ospC. Bacterial burden in skin, inguinal lymph node, heart, bladder and tibiotarsal joint varied between tissues and fluctuated over the course of infection possibly in response to unique cues of each microenvironment. Expression of ospC, when normalized for changes in bacterial load, presented unique profiles in murine tissues at different time points. The inguinal lymph node was infected with a significant B. burgdorferi burden, but showed minimal ospC expression. B. burgdorferi infected skin and heart induced expression of ospC early during infection while the bladder and tibiotarsal joint continued to display PospC driven luminescence throughout the 21 day time course. Localized skin borrelial burden increased dramatically in the first 96 hours following inoculation, which was not paralleled with an increase in ospC expression, despite the requirement of ospC for dermal colonization. Quantitation of bioluminescence representing ospC expression in individual tissues was validated by qRT-PCR of the native ospC transcript. Taken together, the temporal regulation of ospC expression in distal tissues suggests a role for this virulence determinant beyond

  7. [Seroprevalence of Borrelia burgdorferi and tick-borne encephalitis virus in a rural area of Samsun, Turkey].

    Science.gov (United States)

    Aslan Başbulut, Eşe; Gözalan, Ayşegül; Sönmez, Cemile; Cöplü, Nilay; Körhasan, Berrin; Esen, Berrin; Akın, Levent; Ertek, Mustafa

    2012-04-01

    Lyme disease or lyme borreliosis is a zoonosis caused by Borrelia burgdorferi transmitted by ticks, especially Ixodes species. Lyme borreliosis is a multi-systemic disease that invades the skin, musculoskeletal, cardiovascular and central nervous systems. Tick-borne encephalitis (TBE) is an important arboviral infection caused by tick-borne encephalitis virus (TBEV). The central nervous system is affected and the disease most often manifests as meningitis, encephalitis or meningoencephalitis. Previous studies have shown that B.burgdorferi and TBEV can be transmitted by the same tick species (Ixodes ricinus). Although the geographic location and climate is similar to some south-eastern European countries where lyme borreliosis and TBE have been reported, the incidence and prevalence of these diseases in Turkey still remain unclear. The aim of this study was to determine the seroprevelance of B.burgdorferi and TBEV in healthy population in Tekkeköy (41° 8-13' North; 36° 24-31' East), a district of Samsun province, Turkey with evidence of tick-borne disease and to explore the possible correlations of life styles of healthy individuals and prevelance. The cross-sectional study population included 419 people selected using a random proportional sampling method. All participants were asked at interview to complete a questionnaire and peripheral blood samples were collected. From the blood samples, B.burgdorferi IgG and IgM antibodies were evaluated using commercial ELISA (Euroimmun, Germany) and confirmed with Western blot (WB, Euroimmun, Germany). ELISA method was also used to asses IgM and IgG antibodies against TBEV, and neutralization test was used for confirmation. Of the 419 samples, 17 (4%) were positive for B.burgdorferi IgG by ELISA, however 14 (14/419; 3.3%) of them were confirmed by WB. B.burgdorferi seropositivity was higher among people living in rural areas, at an altitude of ≥ 400 meters and in locations ecologically suitable for wild boar and rabbits

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. 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.

  13. Quantitative detection of Borrelia burgdorferi sensu lato in erythema migrans skin lesions using internally controlled duplex real time PCR.

    Science.gov (United States)

    O'Rourke, Maria; Traweger, Andreas; Lusa, Lara; Stupica, Dasa; Maraspin, Vera; Barrett, P Noel; Strle, Franc; Livey, Ian

    2013-01-01

    B. burgdorferi sensu stricto, B. afzelii, B. garinii and B. bavariensis are the principal species which account for Lyme borreliosis (LB) globally. We have developed an internally controlled duplex quantitative real time PCR assay targeting the Borrelia 16S rRNA and the human RNAseP genes. This assay is well-suited for laboratory confirmation of suspected cases of LB and will be used to assess the efficacy of a vaccine against LB in clinical trials. The assay is highly specific, successfully detecting DNA extracted from 83 diverse B. burgdorferi sensu lato strains representing all major species causing LB, while 21 unrelated microbial species and human genomic DNA tested negative. The assay was highly reproducible and sensitive, with a lower limit of detection of 6 copies per PCR reaction. Together with culture, the assay was used to evaluate paired 3 mm skin biopsy samples taken from 121 patients presenting with solitary erythema migrans (EM) lesion. PCR testing identified more positive biopsy samples than culture (77.7% PCR positive versus 55.1% culture positive) and correctly identified all specimens scored as culture positive. OspA-based typing identified the majority of isolates as B. afzelii (96.8%) and the bacterial load was significantly higher in culture positive biopsies than in culture negative biopsies (Phistory of LB (P = 0.10). This is the first quantitative PCR study of human skin biopsies predominantly infected with B. afzelii and the first study to demonstrate a clear relationship between clinical symptoms in B. afzelii-infected patients and Borrelia burden.

  14. 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

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

    DEFF Research Database (Denmark)

    Lebech, Anne-Mette

    2002-01-01

    a histological B. burgdorferi specific immunophosphatase-staining method. The utility of the PCR was then tested for identification of B. burgdorferi DNA in skin biopsies from 31 patients with erythema migrans. The sensitivity of PCR was 71%, which was superior to culture and serology. Based on own and otherwise...... published results there is clear evidence for PCR being the most sensitive and specific test for detection of B. burgdorferi in skin biopsies from patients with both early and late dermatoborreliosis. However, since the clinical diagnosis of dermatoborreliosis in most instances is easy, an invasive...... been studied by phenotypic and genotypic traits and have been shown to be highly heterogeneous. Our first approach was to genotype a panel of human B. burgdorferi isolates by restriction fragment length polymorphism (RFLP) of three genes. Thereafter, sequencing and dideoxy fingerprinting of osp...

  16. Cellular and humoral immune responses to Borrelia burgdorferi antigens in patients with culture-positive early Lyme disease.

    Science.gov (United States)

    Vaz, A; Glickstein, L; Field, J A; McHugh, G; Sikand, V K; Damle, N; Steere, A C

    2001-12-01

    We determined cellular and humoral immune responses to Borrelia burgdorferi lysate and to recombinant flagellin (FlaB), OspC, and OspA in acute- and convalescent-phase samples from 39 culture-positive patients with erythema migrans and in 20 healthy control subjects. During the acute illness, a median of 4 days after the onset of erythema migrans, 51% of the patients had proliferative cellular responses and 72% had antibody responses to at least one of the borrelial antigens tested. During convalescence, at the conclusion of antibiotic therapy, 64% of the patients had proliferative cellular reactivity and 95% had antibody reactivity with at least one of the spirochetal antigens tested. In both acute- and convalescent-phase samples, cellular immune responses were found as frequently to OspA as to OspC and FlaB. Although antibody responses were also frequently seen to OspC and FlaB, only a few patients had marginal antibody reactivity with OspA. The percentage of patients with proliferative responses was similar in those with clinical evidence of localized or disseminated infection, whereas humoral reactivity was found more often in those with disseminated disease. We conclude that cellular and humoral responses to B. burgdorferi antigens are often found among patients with early Lyme disease. In contrast with the other antigens tested, cellular but not humoral reactivity was often found with OspA.

  17. 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

  18. 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.

  19. 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

  20. 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

  1. 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.

  2. Borrelia burgdorferi RST1 (OspC type A) genotype is associated with greater inflammation and more severe Lyme disease.

    Science.gov (United States)

    Strle, Klemen; Jones, Kathryn L; Drouin, Elise E; Li, Xin; Steere, Allen C

    2011-06-01

    Evidence is emerging for differential pathogenicity among Borrelia burgdorferi genotypes in the United States. By using two linked genotyping systems, ribosomal RNA intergenic spacer type (RST) and outer surface protein C (OspC), we studied the inflammatory potential of B. burgdorferi genotypes in cells and patients with erythema migrans or Lyme arthritis. When macrophages were stimulated with 10 isolates of each RST1, RST2, or RST3 strain, RST1 (OspC type A)-stimulated cells expressed significantly higher levels of IL-6, IL-8, chemokine ligand (CCL) 3, CCL4, tumor necrosis factor, and IL-1β, factors associated with innate immune responses. In peripheral blood mononuclear cells, RST1 strains again stimulated significantly higher levels of these mediators. Moreover, compared with RST2, RST1 isolates induced significantly more interferon (IFN)-α, IFN-γ, and CXCL10, which are needed for adaptive immune responses; however, OspC type I (RST3) approached RST1 (OspC type A) in stimulating these adaptive immune mediators. Similarly, serum samples from patients with erythema migrans who were infected with the RST1 genotype had significantly higher levels of almost all of these mediators, including exceptionally high levels of IFN-γ-inducible chemokines, CCL2, CXCL9, and CXCL10; and this pronounced inflammatory response was associated with more symptomatic infection. Differences among genotypes were not as great in patients with Lyme arthritis, but those infected with RST1 strains more often had antibiotic-refractory arthritis. Thus, the B. burgdorferi RST1 (OspC type A) genotype, followed by the RST3 (OspC type I) genotype, causes greater inflammation and more severe disease, establishing a link between spirochetal virulence and host inflammation.

  3. 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.

  4. Quantitative detection of Borrelia burgdorferi sensu lato in erythema migrans skin lesions using internally controlled duplex real time PCR.

    Directory of Open Access Journals (Sweden)

    Maria O'Rourke

    Full Text Available B. burgdorferi sensu stricto, B. afzelii, B. garinii and B. bavariensis are the principal species which account for Lyme borreliosis (LB globally. We have developed an internally controlled duplex quantitative real time PCR assay targeting the Borrelia 16S rRNA and the human RNAseP genes. This assay is well-suited for laboratory confirmation of suspected cases of LB and will be used to assess the efficacy of a vaccine against LB in clinical trials. The assay is highly specific, successfully detecting DNA extracted from 83 diverse B. burgdorferi sensu lato strains representing all major species causing LB, while 21 unrelated microbial species and human genomic DNA tested negative. The assay was highly reproducible and sensitive, with a lower limit of detection of 6 copies per PCR reaction. Together with culture, the assay was used to evaluate paired 3 mm skin biopsy samples taken from 121 patients presenting with solitary erythema migrans (EM lesion. PCR testing identified more positive biopsy samples than culture (77.7% PCR positive versus 55.1% culture positive and correctly identified all specimens scored as culture positive. OspA-based typing identified the majority of isolates as B. afzelii (96.8% and the bacterial load was significantly higher in culture positive biopsies than in culture negative biopsies (P<0.001. The quantitative data also enabled relationships between Borrelia burden and patient symptoms to be evaluated. The bacterial load was significantly higher among patients with systemic symptoms than without (P = 0.02 and was significantly higher for biopsies retrieved from patients with EM lesions with central clearing (P<0.001. 16S copy numbers were moderately lower in samples from patients reporting a history of LB (P = 0.10. This is the first quantitative PCR study of human skin biopsies predominantly infected with B. afzelii and the first study to demonstrate a clear relationship between clinical symptoms in B. afzelii

  5. 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...

  6. 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...

  7. 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.

  8. Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks and assessment of entomological risk index at localities in Belgrade

    Directory of Open Access Journals (Sweden)

    Krstić Milena

    2016-01-01

    Full Text Available Background/Aim. The first case of human Lyme borreliosis (LB in Serbia was recorded in 1987. The number of reported LB cases has increased in the past decade. The aim of this study was to estimate the density of Ixodes ricinus (I. ricinus ticks, the prevalence of Borrelia burgdorferi sensu lato (B. burgdorferi in them, and entomological risk index (ERI at 19 Belgrade localities which were grouped into three categories (forests, parkforests, parks. The values of ERI were compared with the number of tick bites in humans. Methods. Ticks were collected monthly by using the flag hours method and the infection rate was determined by using dark field microscopy. The ERI value was calculated for each locality where the ticks were collected. The related data about tick bites was obtained from the patient protocol of the Institute of Epidemiology, Military Medical Academy, Belgrade. Results. The total number of collected ticks, the number of nymphs and the infection rates of the nymphs were significantly higher in forests (p < 0.05 than park-forests and parks. Statistically, the ERI value was significantly higher in forests than parks of Belgrade (χ2 = 7.78, p < 0.01. In March and July, the ERI value was also significantly higher in forests, than park-forests (p < 0.01 and parks (p < 0.01. May was the month with the highest ERI value in each ecological category (forests p < 0.05; park-forests p < 0.01; parks p < 0.001. However, the number of tick bites in humans did not correlate with ERI values. Conclusion. The obtained results indicate that the risk of tick bite and human exposure to B. burgdorferi sensu lato is present at all selected localities in Belgrade. For a more comprehensive Lyme disease risk assessment the method of entomological risk index assessment should be combined with other methods, taking into consideration all tick stages and the behaviour and habits of people who may get infected B. burgdorferi sensu lato.

  9. 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.

  10. 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

  11. 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 high...

  12. Abundance of Ixodes ricinus and prevalence of Borrelia burgdorferi s.l. in the nature reserve Siebengebirge, Germany, in comparison to three former studies from 1978 onwards

    Directory of Open Access Journals (Sweden)

    Schwarz Alexandra

    2012-11-01

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

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

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    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

  14. 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

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

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    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.

  16. 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.

  17. 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

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

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    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.

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

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    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.

  20. 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...... and a 1.8-fold rise for the C6 assay. Of 1,886 persons, 102/101 (5.4%) had a significant rise in antibody reactivity in the flagellar assay or the C6 assay. Among 40 cases with a diagnosis of Lyme borreliosis, the sensitivities corresponding to a rise in antibodies were 33% and 50% for the flagellar...... in either ELISA but only 40 (2.1%) developed clinical Lyme borreliosis....

  1. 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.

  2. Host, habitat and climate preferences of Ixodes angustus (Acari: Ixodidae) and infection with Borrelia burgdorferi and Anaplasma phagocytophilum in California, USA.

    Science.gov (United States)

    Stephenson, Nicole; Wong, Johnny; Foley, Janet

    2016-10-01

    The Holarctic tick Ixodes angustus is a competent vector for Borrelia burgdorferi, the etiologic agent of Lyme disease, and possibly Anaplasma phagocytophilum, the etiologic agent of granulocytic anaplasmosis, as well. From 2005 to 2013, we collected host-feeding I. angustus individuals from live-trapped small mammals and by flagging vegetation from 12 study sites in northern and central California, and tested for B. burgdorferi sensu lato, A. phagocytophilum, and Rickettsia spp. DNA by real-time PCR. Among 261 I. angustus collected (259 from hosts and two by flagging), the most common hosts were tree squirrels (20 % of ticks) and chipmunks (37 %). The PCR-prevalence for A. phagocytophilum and B. burgdorferi in ticks was 2 % and zero, respectively. The minimum infection prevalence on pooled DNA samples was 10 % for Rickettsia spp. DNA sequencing of the ompA gene identified this rickettsia as Candidatus Rickettsia angustus, a putative endosymbiont. A zero-inflated negative binomial mixed effects model was used to evaluate geographical and climatological predictors of I. angustus burden. When host species within study site and season within year were included in the model as nested random effects, all significant variables revealed that I. angustus burden increased as temperature decreased. Together with published data, these findings suggest that I. angustus is a host generalist, has a broad geographic distribution, is more abundant in areas with lower temperature within it's range, and is rarely infected with the pathogens A. phagocytophilum and B. burgdorferi.

  3. 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

  4. 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.

  5. Anaplasmataceae and Borrelia burgdorferi sensu lato in the sand lizard Lacerta agilis and co-infection of these bacteria in hosted Ixodes ricinus ticks

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    Ekner Anna

    2011-09-01

    Full Text Available Abstract Background Anaplasmataceae and Borrelia burgdorferi s.l. are important tick-borne bacteria maintained in nature by transmission between ticks and vertebrate hosts. However, the potential role of lizards as hosts has not been sufficiently studied. Results The current study showed that 23 of 171 examined sand lizards Lacerta agilis were PCR positive for Anaplasmataceae. The nucleotide sequences of the several selected PCR products showed 100% homology with Anaplasma spp. found in Ixodes ricinus collected in Tunisia and Morocco (AY672415 - AY672420. 1.2% of lizard collar scale samples were PCR positive for B. lusitaniae. In addition, 12 of 290 examined I. ricinus were PCR positive for B. burgdorferi s.l. and 82 were PCR positive for Anaplasmatacea. The number of ticks per lizard and the number of ticks PCR positive for both microorganisms per lizard were strongly correlated. Moreover, we found a significant correlation between numbers of ticks infected with Anaplasmataceae and with B. burgdorferi s.l. living on the same lizard. However, there was no significant correlation between detection of both bacteria in the same tick. Conclusions To the best of our knowledge, this is the first report of Anaplasmataceae DNA and additionally the second report of B. burgdorferi s.l DNA detection in the sand lizard.

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

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    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.

  7. 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 ...

  8. 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...

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

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    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.

  11. Human Lyme arthritis and the immunoglobulin G antibody response to the 37-kilodalton arthritis-related protein of Borrelia burgdorferi.

    Science.gov (United States)

    Salazar, Carlos A; Rothemich, Monika; Drouin, Elise E; Glickstein, Lisa; Steere, Allen C

    2005-05-01

    In Borrelia burgdorferi-infected C3H-scid mice, antiserum to a differentially expressed, 37-kDa spirochetal outer-surface protein, termed arthritis-related protein (Arp), has been shown to prevent or reduce the severity of arthritis. In this study, we determined the immunoglobulin G (IgG) antibody responses to this spirochetal protein in single serum samples from 124 antibiotic-treated human patients with early or late manifestations of Lyme disease and in serial serum samples from 20 historic, untreated patients who were followed longitudinally from early infection through the period of arthritis. These 20 patients were representative of the spectrum of the severity and duration of Lyme arthritis. Among the 124 antibiotic-treated patients, 53% with culture-proven erythema migrans (EM) had IgG responses to recombinant glutathione S-transferase (GST)-Arp, as did 59% of the patients with facial palsy and 68% of those with Lyme arthritis. In addition, 75 to 80% of the 20 past, untreated patients had reactivity with this protein when EM was present, during initial episodes of joint pain, or during the maximal period of arthritis. There was no association at any of these three time points between GST-Arp antibody levels and the severity of the maximal attack of arthritis or the total duration of arthritis. Thus, after the first several weeks of infection, 60 to 80% of patients had IgG antibody responses to GST-Arp, but this response did not correlate with the severity or duration of Lyme arthritis.

  12. 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.

  13. 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.

    Science.gov (United States)

    Hauser, U; Lehnert, G; Wilske, B

    1998-07-01

    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, G. Lehnert, R. Lobentanzer, and B. Wilske, J. Clin. Microbiol. 35:1433-1444, 1997). For the present work, the data bank from that study, containing information about the reactivities of 330 sera (from patients at different stages of LB [n = 189]; control group, n = 141), was reused. The specificities and sensitivities of various combinations of proteins from different strains were calculated for different interpretation criteria. For immunoglobulin G (IgG) WB, the recommended combination of antigens available to date as recombinant proteins included p83/100 of PKa2, p83/100 of PKo, p39 of PKo, p39 of PBi, and OspC of PBi (interpretation criterion, at least one reactive band required for a positive WB; specificity, 96.5%; sensitivity, 56.1%). The further addition of p58 of PKo, p17 of PKo, or p14 of PKo was most favorable in terms of both a considerable gain of sensitivity and little loss of specificity. IgG Western blotting with a whole-cell lysate of strain PKo might be improved by the addition of OspC of PBi. For IgG WB, the best combination, out of all bands, was p83/100, p58, p39, p30, and p21 of all three strains and OspC of PBi, p17b of PBi, p56 of PKa2, p43 of PKo, p17 of PKo, and p14 of PKo (interpretation criterion, at least two reactive bands required for a positive WB; specificity, 97.2%; sensitivity, 61.4%). An interpretation criterion of at least two reactive bands is more reliable than one of only one reactive band. For IgM WB, the best combination was OspC of PKo

  14. 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

  15. 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.

  16. The Borrelia burgdorferi RelA/SpoT Homolog and Stringent Response Regulate Survival in the Tick Vector and Global Gene Expression during Starvation.

    Directory of Open Access Journals (Sweden)

    Dan Drecktrah

    2015-09-01

    Full Text Available As the Lyme disease bacterium Borrelia burgdorferi traverses its enzootic cycle, alternating between a tick vector and a vertebrate host, the spirochete must adapt and persist in the tick midgut under prolonged nutrient stress between blood meals. In this study, we examined the role of the stringent response in tick persistence and in regulation of gene expression during nutrient limitation. Nutritionally starving B. burgdorferi in vitro increased the levels of guanosine tetraphosphate (ppGpp and guanosine pentaphosphate (pppGpp, collectively referred to as (pppGpp, products of the bifunctional synthetase/hydrolase RelBbu (RelA/SpoT homolog. Conversely, returning B. burgdorferi to a nutrient-rich medium decreased (pppGpp levels. B. burgdorferi survival in ticks between the larval and nymph blood meals, and during starvation in vitro, was dependent on RelBbu. Furthermore, normal morphological conversion from a flat-wave shape to a condensed round body (RB form during starvation was dependent on RelBbu; relBbu mutants more frequently formed RBs, but their membranes were compromised. By differential RNA sequencing analyses, we found that RelBbu regulates an extensive transcriptome, both dependent and independent of nutrient stress. The RelBbu regulon includes the glp operon, which is important for glycerol utilization and persistence in the tick, virulence factors and the late phage operon of the 32-kb circular plasmid (cp32 family. In summary, our data suggest that RelBbu globally modulates transcription in response to nutrient stress by increasing (pppGpp levels to facilitate B. burgdorferi persistence in the tick.

  17. Birds as reservoirs for Borrelia burgdorferi s.l. in Western Europe: circulation of B. turdi and other genospecies in bird-tick cycles in Portugal.

    Science.gov (United States)

    Norte, A C; Ramos, J A; Gern, L; Núncio, M S; Lopes de Carvalho, I

    2013-02-01

    Birds are important in the ecology of Borrelia burgdorferi sensu lato (s.l.) because they are important hosts for vector tick immature stages and are known reservoirs for some Borrelia genospecies. The aim of our study was to assess the role of common passerine bird species as reservoirs for B. burgdorferi s.l. in Western Europe. We surveyed birds in enzootic areas in Portugal, where no information is available for birds as reservoirs for this aetiologic agent and where B. lusitaniae, for which few reservoirs have been identified, is the dominant genospecies. Twenty-three birds (2.9%), including Turdus merula, T. philomelos, Parus major and Fringilla coelebs harboured infected ticks, but only Turdus sp. harboured infected tick larvae. In one study area, although B. lusitaniae was dominant in questing Ixodes ricinus, no ticks feeding on birds were infected with this genospecies, and B. valaisiana was the dominant genospecies in I. ricinus larvae feeding on birds. In the other area ticks collected from birds were mainly I. frontalis which were infected with B. turdi. Two skin biopsies (4.2%) from two T. merula were positive, one for B. valaisiana and the other for B. turdi. This is the first report for B. turdi in Western Europe.

  18. 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.

  19. 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...

  20. 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

  1. A proposal for the reliable culture of Borrelia burgdorferi from patients with chronic Lyme disease, even from those previously aggressively treated.

    Science.gov (United States)

    Phillips, S E; Mattman, L H; Hulínská, D; Moayad, H

    1998-01-01

    Since culture of Borrelia burgdorferi from patients with chronic Lyme disease has been an extraordinarily rare event, clarification of the nature of the illness and proving its etiology as infectious have been difficult. A method for reliably and reproducibly culturing B. burgdorferi from the blood of patients with chronic Lyme disease was therefore sought by making a controlled blood culture trial studying 47 patients with chronic Lyme disease. All had relapsed after long-term oral and intravenous antibiotics. 23 patients with other chronic illness formed the control group. Positive cultures were confirmed by fluorescent antibody immuno-electron microscopy using monoclonal antibody directed against Osp A, and Osp A PCR. 43/47 patients (91%) cultured positive. 23/23 controls (100%) cultured negative. Although persistent infection has been, to date, strongly suggested in chronic Lyme disease by positive PCR and antigen capture, there are major problems with these tests. This new method for culturing B. burgdorferi from patients with chronic Lyme disease certainly defines the nature of the illness and establishes that it is of chronic infectious etiology. This discovery should help to reestablish the gold standard in laboratory diagnosis of Lyme disease.

  2. 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).

  3. Serum antibodies against Borrelia afzelii, Borrelia burgdorferi sensu stricto and the 41-kiloDalton flagellin in patients from a Lyme borreliosis endemic area: analysis by EIA and immunoblot.

    Science.gov (United States)

    Nilsson, I; von Rosen, I A

    1996-12-01

    To evaluate the possible importance of antigenic heterogeneity in the serological diagnosis of Lyme borreliosis a study was performed using antigens from various Lyme Borrelia strains. Serum samples from 102 patients with clinical signs of the infection, all living in an endemic area in southern Sweden, were evaluated by four enzyme immuno assays (EIA). The sera were initially tested for the immunoglobulin G response to antigens from a local Borrelia afzelii strain (ACA1). Serum samples from healthy blood donors residing in the same region were used to define seropositivity in the ACAI-EIA. Immunoblotting was performed with the ACAI antigen and the reactive bands were analysed. A serum was defined as positive when at least four of the Borrelia specific polypeptides (OspC, OspA, OspB, p39, p41 [flagellin], p83, p94, 110kDa) were stained. The same sera were then analysed in three other IgG enzyme immunoassays, one based on antigens from Borrelia burgdorferi sensu stricto B31, and another on pooled protein fractions from strains B31 and ACAI. In the third EIA, sera were analysed for antiflagellin reactivity (B, afzelii strain DK-1). An inconstant immune response was demonstrated in the EIAs and the seropositivity varied between 30-47% when low positive values were excluded, and between 38-73% if all values were included. Fifty sera (50/102) met the criteria for a positive immunoblot, but positive immunoblots were detected with both low positive and negative sera independent of antigen used in the EIAs. Antigens of the local B. afzelii strain were found to detect a higher number of seropositive individuals, which suggests that the antibody reactivity to Lyme Borrelia increases when antigens from a strain endemic in a particular geographical region are used. Data from this study suggest that EIA alone seems insufficient for the serodiagnosis, and antigenic heterogeneity of Lyme Borrelia spp. influences the performance of serum antibody tests. The reliability of

  4. 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

  5. Seropositivity of Borrelia burgdorferi in a cohort of symptomatic cats from Europe based on a C6-peptide assay with discussion of implications in disease aetiology.

    Science.gov (United States)

    Pantchev, Nikola; Vrhovec, Majda Globokar; Pluta, Silvia; Straubinger, Reinhard K

    2016-01-01

    There are only few reports on Lyme borreliosis (LB) in cats. The reasons might be a different tick infestation in cats compared to dogs, a low susceptibility for tick-borne infections or a low awareness of veterinarians for tick-borne diseases in feline patients. The aim of this study was to determine the proportion of antibodies against Borrelia burgdorferi sensu lato (Bbsl) in feline sera, to compare the significance of feline versus canine LB, as well as to evaluate possible implications on disease occurrence. Specific antibodies against the C6-peptide of Bbsl in cats were detected by a rapid test based on enzyme immunoassay technique. The serum samples were sent to a diagnostic laboratory by veterinarians from Germany and other European countries with request for Borrelia serology in the years 2009-2011. Veterinarians were asked for information regarding the cats' location, age, gender, clinical signs, treatment and follow-up. In six of 271 (2.2%; 95% CI: 0.8-4.8%) cat sera, antibodies against the C6-peptide of Bbsl were detected. Proportion of Borrelia antibody-positive cat sera was significantly lower than the one determined for dogs during the same time period. All positive cats lived in countries endemic for LB (Germany, Sweden and Belgium), and all C6-antibody positive cats with the exception of one cat showed clinical signs. Possible implications on disease occurrence are discussed. Data presented here demonstrate a lower prevalence of Borrelia specific C6-antibodies in European cats when compared to dogs residing in the same regions. The absence of antibodies against Bbsl in 97.8% (95% CI: 95.2-99.2%) of the submitted samples indicate that diagnosis "feline LB"is rare in cats. Nevertheless, LB should be considered in cats with compatible clinical signs (e.g. shifting leg lameness, to less extent neurological signs) when other differential diagnoses are ruled out. PMID:27529996

  6. Seroprevalence against Borrelia burgdorferi sensu lato and occurence of antibody co-expression with Anaplasma phagocytophilum in dogs in Latvia

    OpenAIRE

    Berzina, Inese; Matise, Ilze

    2013-01-01

    Background Lyme disease is commonly diagnosed in humans in Latvia, but up to date no studies have been performed to investigate its prevalence in dogs. The aim of this study was to evaluate if seroprevalence against B. burgdorferi sensu lato (B. burgdorferi s.l.) and co-expression of antibodies against B.burgdorferi s.l. and A. phagocytophilum is higher in dogs with clinical suspicion of tick-borne diseases compared to healthy dogs. Findings Venous blood was taken from healthy dogs (n=441) an...

  7. Recognition of multiple antibody epitopes throughout Borrelia burgdorferi p66, a candidate adhesin, in patients with early or late manifestations of Lyme disease.

    Science.gov (United States)

    Ntchobo, H; Rothermel, H; Chege, W; Steere, A C; Coburn, J

    2001-03-01

    Antibody responses to p66, a candidate integrin ligand of Borrelia burgdorferi, were studied in 79 patients with early or late manifestations of Lyme disease. The central portion of p66 was previously shown to contain all of the information required for specific recognition of beta3-chain integrins, but work by others had suggested that the C-terminal portion of the protein contains a single surface-exposed, immunodominant loop. In examining antibody responses to full-length p66 and to three overlapping fragments of the protein, we found that the majority of Lyme disease patients had immunoglobulin M (IgM) and/or IgG responses to p66 and that, particularly early in the disease, epitopes throughout p66 were recognized. Among patients with later manifestations of the illness, antibody responses to the C-terminal portion of the protein were more prominent. These results demonstrate that Lyme disease patient sera recognize epitopes throughout p66.

  8. 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.

  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. Comparative population genomics of the Borrelia burgdorferi species complex reveals high degree of genetic isolation among species and underscores benefits and constraints to studying intra-specific epidemiological processes.

    Directory of Open Access Journals (Sweden)

    Maude Jacquot

    Full Text Available Lyme borreliosis, one of the most frequently contracted zoonotic diseases in the Northern Hemisphere, is caused by bacteria belonging to different genetic groups within the Borrelia burgdorferi species complex, which are transmitted by ticks among various wildlife reservoirs, such as small mammals and birds. These features make the Borrelia burgdorferi species complex an attractive biological model that can be used to study the diversification and the epidemiology of endemic bacterial pathogens. We investigated the potential of population genomic approaches to study these processes. Sixty-three strains belonging to three species within the Borrelia burgdorferi complex were isolated from questing ticks in Alsace (France, a region where Lyme disease is highly endemic. We first aimed to characterize the degree of genetic isolation among the species sampled. Phylogenetic and coalescent-based analyses revealed clear delineations: there was a ∼50 fold difference between intra-specific and inter-specific recombination rates. We then investigated whether the population genomic data contained information of epidemiological relevance. In phylogenies inferred using most of the genome, conspecific strains did not cluster in clades. These results raise questions about the relevance of different strategies when investigating pathogen epidemiology. For instance, here, both classical analytic approaches and phylodynamic simulations suggested that population sizes and migration rates were higher in B. garinii populations, which are normally associated with birds, than in B. burgdorferi s.s. populations. The phylogenetic analyses of the infection-related ospC gene and its flanking region provided additional support for this finding. Traces of recombination among the B. burgdorferi s.s. lineages and lineages associated with small mammals were found, suggesting that they shared the same hosts. Altogether, these results provide baseline evidence that can be used

  11. 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。

  12. Analysis of the intrathecal immune response in neuroborreliosis to a sonicate antigen and three recombinant antigens of Borrelia burgdorferi sensu stricto.

    Science.gov (United States)

    Kaiser, R; Rauer, S

    1998-03-01

    The intrathecal synthesis of borrelial-specific IgM- and IgG-antibodies was studied in 67 patients with neuroborreliosis and in 14 patients with neurosyphilis (controls). Antibody concentrations in serum and in the cerebrospinal fluid were determined by an enzyme immunoassay (EIA) using, as antigens, a sonicate of Borrelia burgdorferi, the recombinant 14 kDa flagellin fragment, the outer surface protein C (22 kDa), and the high molecular mass protein p83 (83 kDa). In the sonicate EIA, IgG- and/or IgM-antibodies to Borrelia burgdorferi in serum were detected in all patients with neuroborreliosis and in 71% of patients with neurosyphilis. Intrathecal synthesis of borrelial-specific IgG- and/or IgM-antibodies was demonstrated in 82% of patients with neuroborreliosis and in 71% of patients with neurosyphilis. Immunoglobulin G- and/or IgM-antibodies in serum against any of the recombinant antigens were detected in 92% of patients with neuroborreliosis and in none of those with neurosyphilis. Intrathecal synthesis of IgG- and/or IgM-antibodies to individual recombinant antigens was demonstrated in 67% of patients with neuroborreliosis and in none of those with neurosyphilis. The sensitivity of the recombinant antigens in serum was almost equal to that of the sonicate EIA, whereas the recombinant antigens were clearly less sensitive in the estimation of the intrathecal specific immune response. It was concluded that in suspected cases of neuroborreliosis, the estimation of high specific antibodies in the recombinant EIA will be helpful in confirming the diagnosis.

  13. Wild turkey (Meleagris gallopavo) as a host of ixodid ticks, lice, and Lyme disease spirochetes (Borrelia burgdorferi sensu lato) in California state parks.

    Science.gov (United States)

    Lane, Robert S; Kucera, Thomas F; Barrett, Reginald H; Mun, Jeomhee; Wu, Chunling; Smith, Vincent S

    2006-10-01

    Rio Grande wild turkeys (Meleagris gallopavo intermedia) were evaluated as potential hosts of ixodid ticks, lice, and Lyme disease spirochetes (Borrelia burgdorferi sensu lato [s.l.]) in three state parks in Sonoma County, California, USA, during 2003 and 2004. In total, 113 birds were collected, 50 (44.2%) of which were found to be infested by 361 ixodid ticks representing three species: the western black-legged tick (Ixodes pacificus, n=248), the rabbit tick (Haemaphysalis leporispalustris, n=112), and one American dog tick (Dermacentor variabilis). Year-round the prevalence of all ticks combined was unrelated to the age or sex of turkeys, and the prevalence of infestation by I. pacificus (35.4%) was significantly higher than it was for either H. leporispalustris (14.2%) or D. variabilis (0.9%). The proportion of the two prevalent tick species differed significantly by life stage with 86.3% of the I. pacificus and 82.1% of the H. leporispalustris enumerated being nymphs and larvae, respectively. Three species of lice were collected, including the chicken body louse Menacanthus stramineus (12.5% of total), Chelopistes meleagridis (37.5% of total), and Oxylipeurus polytrapezius (50% of total). The records for all three ticks are the first ever from wild turkeys, and those for the lice are the first from this host in the far-western United States. Wild turkeys potentially were exposed to the feeding activities of I. pacificus nymphs infected with B. burgdorferi s.l. as 15% of host-seeking nymphs (n=200) collected in woodlands used by turkeys as roosting or foraging areas were infected mainly with B. burgdorferi sensu stricto (s.s.). However, only one (1%) of 90 turkey blood specimens tested by PCR contained B. burgdorferi s.s., and four in vitro, complement-protein assays demonstrated that domestic turkey serum is moderately bacteriolytic for this spirochete. Taken together, these findings indicate that wild turkeys are important avian hosts of I. pacificus nymphs

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

    Directory of Open Access Journals (Sweden)

    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

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

    Directory of Open Access Journals (Sweden)

    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

  16. Antigens of Borrelia burgdorferi recognized during Lyme disease. Appearance of a new immunoglobulin M response and expansion of the immunoglobulin G response late in the illness.

    Science.gov (United States)

    Craft, J E; Fischer, D K; Shimamoto, G T; Steere, A C

    1986-01-01

    Using immunoblots, we identified proteins of Borrelia burgdorferi bound by IgM and IgG antibodies during Lyme disease. In 12 patients with early disease alone, both the IgM and IgG responses were restricted primarily to a 41-kD antigen. This limited response disappeared within several months. In contrast, among six patients with prolonged illness, the IgM response to the 41-kD protein sometimes persisted for months to years, and late in the illness during arthritis, a new IgM response sometimes developed to a 34-kD component of the organism. The IgG response in these patients appeared in a characteristic sequential pattern over months to years to as many as 11 spirochetal antigens. The appearance of a new IgM response and the expansion of the IgG response late in the illness, and the lack of such responses in patients with early disease alone, suggest that B. burgdorferi remains alive throughout the illness. Images PMID:3531237

  17. 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

  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. 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

  20. 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...

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    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. A phylogenomic and molecular marker based proposal for the division of the genus Borrelia into two genera: the emended genus Borrelia containing only the members of the relapsing fever Borrelia, and the genus Borreliella gen. nov. containing the members of the Lyme disease Borrelia (Borrelia burgdorferi sensu lato complex).

    Science.gov (United States)

    Adeolu, Mobolaji; Gupta, Radhey S

    2014-06-01

    The genus Borrelia contains two groups of organisms: the causative agents of Lyme disease and their relatives and the causative agents of relapsing fever and their relatives. These two groups are morphologically indistinguishable and are difficult to distinguish biochemically. In this work, we have carried out detailed comparative genomic analyses on protein sequences from 38 Borrelia genomes to identify molecular markers in the forms of conserved signature inserts/deletions (CSIs) that are specifically found in the Borrelia homologues, and conserved signature proteins (CSPs) which are uniquely present in Borrelia species. Our analyses have identified 31 CSIs and 82 CSPs that are uniquely shared by all sequenced Borrelia species, providing molecular markers for this group of organisms. In addition, our work has identified 7 CSIs and 21 CSPs which are uniquely found in the Lyme disease Borrelia species and eight CSIs and four CSPs that are specific for members of the relapsing fever Borrelia group. Additionally, 38 other CSIs, in proteins which are uniquely found in Borrelia species, also distinguish these two groups of Borrelia. The identified CSIs and CSPs provide novel and highly specific molecular markers for identification and distinguishing between the Lyme disease Borrelia and the relapsing fever Borrelia species. We also report the results of average nucleotide identity (ANI) analysis on Borrelia genomes and phylogenetic analysis for these species based upon 16S rRNA sequences and concatenated sequences for 25 conserved proteins. These analyses also support the distinctness of the two Borrelia clades. On the basis of the identified molecular markers, the results from ANI and phylogenetic studies, and the distinct pathogenicity profiles and arthropod vectors used by different Borrelia spp. for their transmission, we are proposing a division of the genus Borrelia into two separate genera: an emended genus Borrelia, containing the causative agents of relapsing

  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. 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.

  6. Association of treatment-resistant chronic Lyme arthritis with HLA-DR4 and antibody reactivity to OspA and OspB of Borrelia burgdorferi.

    Science.gov (United States)

    Kalish, R A; Leong, J M; Steere, A C

    1993-01-01

    Chronic Lyme arthritis that is unresponsive to antibiotic therapy is associated with an increased frequency of the HLA-DR4 specificity. To determine whether the immune response to a particular polypeptide of Borrelia burgdorferi may be associated with treatment-resistant chronic Lyme arthritis, we correlated the clinical courses and HLA-DR specificities of 128 patients with Lyme disease with their antibody responses to spirochetal polypeptides. Antibody reactivity was determined by Western blotting (immunoblotting) with sonicated whole B. burgdorferi and recombinant forms of its outer surface proteins, OspA and OspB, as the antigen preparations. Of 15 patients monitored for 4 to 12 years, 11 (73%) developed strong immunoglobulin G responses to both OspA and OspB near the beginning of prolonged episodes of arthritis, from 5 months to 7 years after disease onset. When single serum samples from 80 patients with Lyme arthritis, were tested, 57 (71%) showed antibody reactivity to recombinant Osp proteins; in contrast, none of 43 patients who had erythema migrans or Lyme meningitis (P < 0.00001) and 1 of 5 patients who had chronic neuroborreliosis but who never had arthritis (P = 0.03) showed antibody reactivity to these proteins. Among the 60 antibiotic-treated patients with Lyme arthritis, those with the HLA-DR4 specificity and Osp reactivity had arthritis for a significantly longer time after treatment than those who lacked Osp reactivity (median duration, 9.5 versus 4 months; P = 0.009); a similar trend was found for the HLA-DR2 specificity. For other HLA-DR specificities, arthritis resolved within a median duration of 2 months in both Osp-reactive and nonreactive patients. We conclude that the combination of the HLA-DR4 specificity and OspA or OspB reactivity is associated with chronic arthritis and the lack of a response to antibiotic therapy. Images PMID:7685738

  7. Humoral immune response to outer surface protein C of Borrelia burgdorferi in Lyme disease: role of the immunoglobulin M response in the serodiagnosis of early infection.

    Science.gov (United States)

    Fung, B P; McHugh, G L; Leong, J M; Steere, A C

    1994-08-01

    We determined the humoral immune response to outer surface protein C (OspC) of Borrelia burgdorferi in patients with early or late manifestations of Lyme disease and investigated the use of this antigen in the serodiagnosis of early infection. The ospC gene from the low-passage human isolate 297, a North American B. burgdorferi strain, was used to make a recombinant maltose-binding protein (MBP)-OspC fusion protein for serologic tests. This gene showed 84 to 85% nucleotide sequence identity and 76 to 79% amino acid identity with ospC of B. burgdorferi B31 and 2591. The antibody responses to MBP-OspC were determined in serial sera from 15 patients with Lyme disease who were monitored for 4 to 12 years of illness, in single-serum samples from 189 patients with early or late manifestations of the disorder, and in serum samples from 106 control patients. Early in the infection, patients with erythema migrans or meningitis commonly had weak to strong immunoglobulin M (IgM) responses to OspC and sometimes weak to moderate IgG responses. Months to years later, weak to strong IgG reactivity with this protein was often apparent in patients with arthritis, but this response was weak or absent in patients with chronic neuroborreliosis. When acute- and convalescent-phase serum samples from patients with erythema migrans were tested for reactivity against MBP-OspC, the sensitivity of the IgM test was 73% and the specificity was 98%, with either enzyme-linked immunosorbent assay (ELISA) or Western blotting. We conclude that the majority of patients with Lyme disease have a prominent IgM response to OspC early in the illness, which is often followed by a prominent IgG response in patients with arthritis. For the serodiagnosis of early infection, the sensitivity and specificity of IgM ELISA and Western blotting were comparable or slightly improved when MBP-OspC was used as the antigen compared with tests in which spirochetal lysates were used.

  8. 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.

  9. 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

  10. Occurrence of ticks and prevalence of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. in three types of urban biotopes: forests, parks and cemeteries.

    Science.gov (United States)

    Hornok, Sándor; Meli, Marina L; Gönczi, Enikő; Halász, Edina; Takács, Nóra; Farkas, Róbert; Hofmann-Lehmann, Regina

    2014-10-01

    The aim of the present study was to compare different urban biotopes for the occurrence of ixodid tick species, for the population density of Ixodes ricinus and for the prevalence rates of two emerging, zoonotic pathogens. Altogether 2455 ticks were collected from the vegetation on 30 places (forests, parks, cemeteries) of Budapest, Hungary. I. ricinus and Haemaphysalis concinna were collected in all three biotope types, but Dermacentor reticulatus only in parks and forests, and D. marginatus only in a forest. Highest population density of I. ricinus was observed in neglected parts of cemeteries. In females of this tick species the prevalence rates of Anaplasma phagocytophilum and Borrelia burgdorferi s.l. were significantly lower in cemeteries, than in parks or forests. In conclusion, risks associated with the presence of ticks and tick-borne pathogens may be high in a city, but this depends on biotope types, due to habitat-related differences in the vegetation, as well as in the availability of tick hosts and pathogen reservoirs.

  11. 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

  12. 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.

  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. Sensitive and specific serodiagnosis of Lyme disease by enzyme-linked immunosorbent assay with a peptide based on an immunodominant conserved region of Borrelia burgdorferi vlsE.

    Science.gov (United States)

    Liang, F T; Steere, A C; Marques, A R; Johnson, B J; Miller, J N; Philipp, M T

    1999-12-01

    VlsE, the variable surface antigen of Borrelia burgdorferi, contains an immunodominant conserved region named IR(6). In the present study, the diagnostic performance of a peptide enzyme-linked immunosorbent assay (ELISA) based on a 26-mer synthetic peptide (C(6)) with the IR(6) sequence was explored. Sensitivity was assessed with serum samples (n = 210) collected from patients with clinically defined Lyme disease at the acute (early localized or early disseminated disease), convalescent, or late disease phase. The sensitivities for acute-, convalescent-, and late-phase specimens were 74% (29 of 39), 85 to 90% (34 of 40 to 35 of 39), and 100% (59 of 59), respectively. Serum specimens from early neuroborreliosis patients were 95% positive (19 of 20), and those from an additional group of patients with posttreatment Lyme disease syndrome yielded a sensitivity of 62% (8 of 13). To assess the specificity of the peptide ELISA, 77 serum samples from patients with other spirochetal or chronic infections, autoimmune diseases, or neurologic diseases and 99 serum specimens from hospitalized patients in an area where Lyme disease is not endemic were examined. Only two potential false positives from the hospitalized patients were found, and the overall specificity was 99% (174 of 176). Precision, which was assessed with a panel of positive and negative serum specimens arranged in blinded duplicates, was 100%. Four serum samples with very high anti-OspA antibody titers obtained from four monkeys given the OspA vaccine did not react with the C(6) peptide. This simple, sensitive, specific, and precise ELISA may contribute to alleviate some of the remaining problems in Lyme disease serodiagnosis. Because of its synthetic peptide base, it will be inexpensive to manufacture. It also will be applicable to serum specimens from OspA-vaccinated subjects.

  16. 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.

  17. 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.

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

    Science.gov (United States)

    Sirmarová, Jana; Tichá, Lucie; Golovchenko, Marina; Salát, Jiří; Grubhoffer, Libor; Rudenko, Nataliia; Nowotny, Norbert; Růžek, Daniel

    2014-09-01

    This study was conducted to evaluate the prevalence of antibodies against Borrelia bugdorferi (Bb) s.l. and tick-borne encephalitis virus (TBEV) in zoo animals in the Czech Republic. We collected 133 serum samples from 69 animal species from 5 zoos located in different parts of the country. The samples were obtained from even-toed ungulates (n=78; 42 species), odd-toed ungulates (n=32; 11 species), carnivores (n=13; 9 species), primates (n=2, 2 species), birds (n=3; 2 species), and reptiles (n=5; 3 species). A high antibody prevalence (60%) was observed for Bb s.l. On the other hand, only two animals had TBEV-specific antibodies: a markhor (Capra falconeri) and a reindeer (Rangifer tarandus), both from the same zoo, located in an area endemic for TBEV. Both of these animals were also positive for Bb s.l. antibodies. Our results indicate that a high number of animal species in the Czech zoos were exposed to Bb s.l. and that TBEV infection occurred at least in one of the investigated zoos. Considering the pathogenic potential of these two tick-borne pathogens, clinical and serological monitoring should be continued, and therapeutic and preventive measures should be taken when necessary.

  19. 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

  20. 伯氏疏螺旋体传播过程中病原体、媒介、宿主间的相互作用%The pathogen-vector-host interactions during Borrelia burgdorferi transmission

    Institute of Scientific and Technical Information of China (English)

    宝福凯; Erol Fikrig

    2008-01-01

    Lyme borreliosis is a prevalent arthropod-borne disease caused by the spirochete,Borrelia burgdorferi.The microbe consists in nature through an intricate tick-mammal cycle.B.burgdorferi is transmitted to vertebrates via ticks belonging to the Ixodes ricinus complex.Over the last two decades,scientists have been trying to unravel the complex mechanisms by which B.burgdorferi maintained in a unique enzootic cycle.Our special attention has been paid to addressing the molecular interaction strategies that B.burgdorferi employs for effective colonization,migration and transmission through ticks.Studies have shown that B.burgdorferi expresses a select set of genes in distinct phases of its life cycle- and in specific tissue locations.The outer surface protein(Osp)A of B.burgdorferi is down-regulated within a mammalian host,turned on as soon as the spirochete enters and resides in the arthropod vector.OspA acts as an adhesion,which binds a receptor in the tick miagut,and OspA is required for spirochetes to successfully colonize in the tick midgut.B.burgdorferi lacking OspA cannot survive in the tick.To further understand the mechanism of OspA-based adherence,we have recently identified and characterized the tick receptor for OspA,named TROSPA.TROSPA predominantly localized in the tick midgut,specifically bound to ospA expressing B.burgdorferi and TROSPA knockdown ticks allowed poor attachment of spirochetes in the tick midgut.These observations clearly indicate that like OspA.TROSPA is also actively engaged in the colonization of spirochetes in Ixodes ticks.The OspA-TROSPA interaction is the first illustration of a molecular interface where both microbial as well as vector gene products equally contribute to B.burgdorferi survival in nature,We also find that there are the complex pathogen vector-mammalian host triangle interactions during mammalian infection tith B.burgdorferi.Lxodes tick salivary protein Salp15 help spirochetes to infect the mice.B.burgdorferi basic

  1. 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

  2. Serodiagnosis of Lyme disease by kinetic enzyme-linked immunosorbent assay using recombinant VlsE1 or peptide antigens of Borrelia burgdorferi compared with 2-tiered testing using whole-cell lysates.

    Science.gov (United States)

    Bacon, Rendi Murphree; Biggerstaff, Brad J; Schriefer, Martin E; Gilmore, Robert D; Philipp, Mario T; Steere, Allen C; Wormser, Gary P; Marques, Adriana R; Johnson, Barbara J B

    2003-04-15

    In a study of US patients with Lyme disease, immunoglobulin (Ig) G and IgM antibody responses to recombinant Borrelia burgdorferi antigen VlsE1 (rVlsE1), IgG responses to a synthetic peptide homologous to a conserved internal sequence of VlsE (C6), and IgM responses to a synthetic peptide comprising the C-terminal 10 amino acid residues of a B. burgdorferi outer-surface protein C (pepC10) were evaluated by kinetic enzyme-linked immunoassay. At 99% specificity, the overall sensitivities for detecting IgG antibody to rVlsE1 or C6 in samples from patients with diverse manifestations of Lyme disease were equivalent to that of 2-tiered testing. When data were considered in parallel, 2 combinations (IgG responses to either rVlsE1 or C6 in parallel with IgM responses to pepC10) maintained high specificity (98%) and were significantly more sensitive than 2-tiered analysis in detecting antibodies to B. burgdorferi in patients with acute erythema migrans. In later stages of Lyme disease, the sensitivities of the in parallel tests and 2-tiered testing were high and statistically equivalent.

  3. 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 (

  4. Early and late antibody responses to full-length and truncated constructs of outer surface protein A of Borrelia burgdorferi in Lyme disease.

    Science.gov (United States)

    Kalish, R A; Leong, J M; Steere, A C

    1995-06-01

    The immunoglobulin G (IgG) antibody response to outer surface protein A (OspA) of Borrelia burgdorferi has been reported to occur late in the course of Lyme disease. To learn when reactivity to particular epitopes of OspA develops and whether the strength of particular responses correlates with the duration of arthritis and HLA-DR specificities, we determined the IgM and IgG responses by enzyme-linked immunosorbent assay in 128 patients with various manifestations of Lyme disease to full-length recombinant OspA and three OspA fragments which divided the protein approximately into thirds. Among the 10 patients who were followed serially, an early IgM response was often found to epitopes in all three fragments of OspA, sometimes accompanied by a weak IgG response, primarily to an epitope in the middle third of the protein. Months to years later, the seven patients who had prolonged or moderate episodes of arthritis developed strong IgG responses to OspA, especially to epitopes in the N-terminal and C-terminal fragments, that paralleled the course of the arthritis. In single serum samples from 128 patients, a similar pattern of IgM and IgG reactivity with OspA epitopes was seen in patients with early or late manifestations of the illness. Of the 80 patients with arthritis, 62 (78%) had IgG responses to OspA, usually with the strongest reactivity to the C-terminal fragment. In these patients, the strength of the IgG response to OspA correlated with the duration of arthritis; in HLA-DR4-positive patients, most of whom had chronic arthritis, this association was attributable to reactivity with the C-terminal fragment. Thus, patients with Lyme disease often have early responses to OspA, but those with prolonged arthritis do not develop IgG responses to certain epitopes of the protein until late in the illness. In patients with HLA-DR4, the strength of IgG reactivity with one or more epitopes in the C-terminal fragment of OspA correlates with the duration of arthritis.

  5. [Ticks Ixodes ricinus (Linnaeus, 1758) from selected municipal forests of the city Poznań and their infection with the spirochetes Borrelia burgdorferi senso lato].

    Science.gov (United States)

    Nowosad, A; Jenek, J; Głazaczow, A; Wal, M

    1999-01-01

    The results of research (1997-1998) on the occurrence of the spirochetes Borrelia burgdorferi s.l. in ticks Ixodes ricinus (L.) in selected municipal forests of the city of Poznań (localities--districts of Poznań: 1 Debiec, 2 Marcelin, 3 Golecin and Wola, 4 Krzyzowniki and Smochowice, 5 Kiekrz and Strzeszynek, 6 Morasko, 7 Piatkowo, 8 Umultowo, 9 Naramowice and Rózany Młyn, 10 Malta and Antoninek) are presented. A total of 1432 ticks were collected from 10 localities (748 larvae, 590 nymphs, 47 males, and 47 females). Out of this number, 266 specimens were selected at random for further analysis (20 larvae, 160 nymphs, 44 males, and 42 females) which were then tested for the presence of the spirochetes using the PCR technique (tab. I-III, fig. 1). Spirochetes were found in 60 specimens (22.6%). Percentages of infected nymphs and males were similar: 25.6% and 27.3%, respectively. The level of infection of females was lower (14.3%) and of larvae lowest (5.0%). Infected ticks were found in all ten localities, but their proportions differed from site to site and varied from 9.5% (Piatkowo) to 34.6% (Krzyzowniki and Smochowice). Special attention was devoted to those municipal woods where ticks were abundant (Kiekrz and Strzeszynek, as well as Malta and Antoninek, and relatively numerous Debiec) and where their extensiveness of infestation was high (Krzyzowniki and Smochowice: 34.6%; Debiec: 29.6%; Naramowice and Rózany Młyn: 28.6%; Malta and Antoninek: 28.1%; and Golecin and Wola: 26.9%). After summing up all the data from the literature on Wielkopolska and the city of Poznań, the mean extensiveness of infestation in Wielkopolska turns out to be 21.8% and in Poznań slightly higher, 22.2% (tab. IV). The infection levels of the particular developmental stages are different, though: highly variable in nymphs (8.3-25.6%), and more stable in males (22.0-27.3%) and females (22.0-24.5%).

  6. 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.

  7. 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

  8. 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.

  9. 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

  10. Use of a hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin and part of the 83-kDa protein as antigen for serodiagnosis of Lyme disease.

    Science.gov (United States)

    Rasiah, C; Rauer, S; Gassmann, G S; Vogt, A

    1994-04-01

    A hybrid protein consisting of the variable region of the Borrelia burgdorferi flagellin (an 18-kDa fragment) and a 59-kDa fragment (lacking the N-terminal part) of the 83-kDa protein has been constructed by genetic engineering. It was expressed as a nonfusion protein of an apparent molecular weight of 77,000 in Escherichia coli. The suitability of this new antigen for the diagnosis of Lyme disease was tested by immunoblotting; for comparison, the recombinant variable region of the flagellin, the 18-kDa fragment (p18), and the whole recombinant 83-kDa protein (p83), both expressed in E. coli, were used. A total of 120 serum samples from various stages of Lyme disease, which were positive in two serological assays, a passive hemagglutination assay and an indirect immunofluorescence assay, were tested. By indirect immunofluorescence, 74 samples were positive for immunoglobulin G (IgG) antibodies and 72 were positive for IgM antibodies. Of these serum samples, 69 of 74 (93%) contained IgG antibodies against p18 and/or p83, and IgG antibodies were detected by the hybrid protein in 67 (90%) samples. IgM antibodies against p18 and/or p83 were detected in 60 of 72 (83%) serum samples, and 57 (79%) serum samples were reactive with the hybrid protein. Twenty serum samples of patients with a history of syphilis and 40 serum samples, negative in routine B. burgdorferi serology, were tested as controls. The hybrid protein, made up of specific epitopes of an early (p18) and late (p83) antigen, is recognized by almost the same number of patient serum samples as the individual antigens.

  11. Strong IgG antibody responses to Borrelia burgdorferi glycolipids in patients with Lyme arthritis, a late manifestation of the infection.

    Science.gov (United States)

    Jones, Kathryn L; Seward, Robert J; Ben-Menachem, Gil; Glickstein, Lisa J; Costello, Catherine E; Steere, Allen C

    2009-07-01

    In this study, the membrane lipids of B. burgdorferi were separated into 16 fractions; the components in each fraction were identified, and the immunogenicity of each fraction was determined by ELISA using sera from Lyme disease patients. Only the 2 glycolipids, acylated cholesteryl galactoside (ACG, BbGL-I) and monogalactosyl diacylglycerol (MgalD, BbGL-II), were immunogenic. Early in the infection, 24 of 84 patients (29%) who were convalescent from erythema migrans and 19 of the 35 patients (54%) with neuroborreliosis had weak IgG responses to purified MgalD, and a smaller percentage of patients had early responses to synthetic ACG. However, almost all of 75 patients with Lyme arthritis, a late disease manifestation, had strong IgG reactivity with both glycolipids. Thus, almost all patients with Lyme arthritis have strong IgG antibody responses to B. burgdorferi glycolipid antigens.

  12. 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....

  13. 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.

  14. 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

    Directory of Open Access Journals (Sweden)

    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

  15. 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.

  16. 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.

  17. Borrelia infection and risk of non-Hodgkin lymphoma

    DEFF Research Database (Denmark)

    Schollkopf, C.; Melbye, M.; Munksgaard, L.;

    2008-01-01

    Reports of the presence of Borrelia burgdorferi DNA in malignant lymphomas have raised the hypothesis that infection with B. burgdorferi may be causally related to non-Hodgkin lymphoma (NHL) development. We conducted a Danish-Swedish case-control study including 3055 NHL patients and 3187.......9-2.0]). However, in analyses of NHL subtypes, self-reported history of B. burgdorferi infection (OR = 2.5 [1.2-5.1]) and seropositivity for anti-Borrelia antibodies (OR = 3.6 [1.8-7.4]) were both associated with risk of mantle cell lymphoma. Notably, this specific association was also observed in persons who did...... not recall Borrelia infection yet tested positive for anti-Borrelia antibodies (OR = 4.2 [2.0-8.9]). Our observations suggest a previously unreported association between B. burgdorferi infection and risk of mantle cell lymphoma Udgivelsesdato: 2008/6/15...

  18. Analysis of Borrelia burgdorferi Genotypes in Patients with Lyme Arthritis: High Frequency of RST 1 Strains in Antibiotic-Refractory Arthritis

    Science.gov (United States)

    Jones, Kathryn L.; McHugh, Gail A.; Glickstein, Lisa J.; Steere, Allen C.

    2009-01-01

    Objective Most of the B. burgdorferi genotypes have been isolated from erythema migrans (EM) skin lesions in patients with Lyme disease; outer-surface protein C (OspC) type K strains, which are 16S-23S rRNA intergenic spacer type 2 (RST 2), are most commonly recovered, but a higher percentage of OspC type A strains (RST 1), the next most common type, are detectable in blood. Our goals were to determine the B. burgdorferi genotypes in the joints of patients with Lyme arthritis. Methods Joint fluid samples from 124 patients seen over a 30-year period were analyzed for OspC types by semi-nested PCR and sequencing, and for RST by nested PCR and RFLP techniques. This information was correlated with clinical outcome. Results OspC and RST genotypes could be determined in 49 of the 124 joint fluid samples (40%). Of the 49 samples, 21 (43%) were OspC type K (RST 2), 11 (22%) were type A (RST 1), and 17 (35%) were distributed among 8 other OspC types and all 3 RSTs. However, among 17 patients who received current antibiotic regimens, all 7 infected with RST 1 strains had antibiotic-refractory arthritis compared with 4 of 6 patients infected with RST 2 strains and only 1 of 4 infected with RST 3 strains (P=0.03). Conclusions Most of the B. burgdorferi genotypes infected the joints of patients with Lyme arthritis, particularly OspC type K (RST 2); and genotype frequencies reflected those in EM skin lesions. However, RST 1 strains were most frequent in patients with antibiotic-refractory arthritis. PMID:19565522

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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

  5. 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.

  6. 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

  7. 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

  8. 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 ...

  9. 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.

  10. 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

    目的为莱姆病血清学诊断和基因工程亚单位疫苗研制提供靶抗原。方法采用377型DNA自动测序仪对莱姆病螺旋体重组质粒pBX1的插入片段进行DNA序列测定,并通过计算机软件对其进行限制性内切酶酶谱分析。然后将重组质粒pBX1在大肠杆菌中进行诱导表达,并对其表达产物进行免疫印迹分析。结果①重组质粒pBX1插入片段大小为477bp,其核苷酸序列与文献报道的p83基因全序列相应区段相比较仅有一个碱基的变异,②成功绘制了该插入片段的限制性内切酶酶谱;③重组质粒在大肠杆菌中诱导表达后获得了29kd的融合蛋白;④Western-blotting分析表明该融合蛋白能与莱姆病多价抗血清呈强阳性印迹反应。结论该研究成功地对莱姆病螺旋体83kd抗原蛋白特异性区段进行了基因重组和表达,为进一步研究其在莱姆病血清学诊断和基因工程亚单位疫苗研制中的应用奠定了基础。%Objective This study was to provide the target antigen for the development of a Lyme disease vaccine and serodiagnosis reagent.Methods We used the automatic DNA sequencing machine (Model 377) to detect the nucleotide sequence of the inserted part of the recombinant plasmid pBX1 from Borrelia burgdorferi B31 strain. The restriction enzyme map of the inserted part of pBX1 was analysed by using computer software. The expressed product of pBX1 in E.coli XLI-Blue MRF′ was analysed by using SDS-PAGE and western-blotting. Results ①DNA sequencing showed that pBX1 contained a 477bp inserted gene fragment,and when it was compared with the published sequence of the specific region of the gene of the 83kd antigen protein from Borrelia burgdorferi B31 strain,only one amino acid codon was different.②The restriction enzyme map of the inserted part of pBX1 was successfully constructed.③The recombinant plasmid pBX1 expressed a 29kd fusion protein in E.coli XL1-Blue MRF

  11. Borrelia species induce inflammasome activation and IL-17 production through a caspase-1-dependent mechanism

    NARCIS (Netherlands)

    Oosting, M.; Veerdonk, F.L. van de; Kanneganti, T.D.; Sturm, P.D.J.; Verschueren, I.; Berende, A.; Meer, J.W. van der; Kullberg, B.J.; Netea, M.G.; Joosten, L.A.B.

    2011-01-01

    Borrelia burgdorferi spirochetes cause Lyme disease, which can result in severe clinical symptoms such as multiple joint inflammation and neurological disorders. IFN-gamma and IL-17 have been suggested to play an important role in the host defense against Borrelia, and in the immunopathology of Lyme

  12. 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...

  13. 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

  14. 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...

  15. 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

  16. 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...

  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. 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.

  1. 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

  2. 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.

  3. 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.

  4. 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.

  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. Evidence of Borrelia autoimmunity-induced component of Lyme carditis and arthritis.

    Science.gov (United States)

    Raveche, Elizabeth S; Schutzer, Steven E; Fernandes, Helen; Bateman, Helen; McCarthy, Brian A; Nickell, Steven P; Cunningham, Madeleine W

    2005-02-01

    We investigated the possibility that manifestations of Lyme disease in certain hosts, such as arthritis and carditis, may be autoimmunity mediated due to molecular mimicry between the bacterium Borrelia burgdorferi and self-components. We first compared amino acid sequences of Streptococcus pyogenes M protein, a known inducer of antibodies that are cross-reactive with myosin, and B. burgdorferi and found significant homologies with OspA protein. We found that S. pyogenes M5-specific antibodies and sera from B. burgdorferi-infected mice reacted with both myosin and B. burgdorferi proteins by Western blots and enzyme-linked immunosorbent assay. To investigate the relationship between self-reactivity and the response to B. burgdorferi, NZB mice, models of autoimmunity, were infected. NZB mice infected with B. burgdorferi developed higher degrees of joint swelling and higher anti-B. burgdorferi immunoglobulin M cross-reactive responses than other strains with identical major histocompatibility complex (DBA/2 and BALB/c). These studies reveal immunological cross-reactivity and suggest that B. burgdorferi may share common epitopes which mimic self-proteins. These implications could be important for certain autoimmunity-susceptible individuals or animals who become infected with B. burgdorferi.

  7. 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.

  8. 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.

  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. 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

  11. 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.

  12. 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

  13. 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

  14. 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...

  15. 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....

  16. 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.

  17. 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.

  18. 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

  19. 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 ...

  20. 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...

  1. Complete genome sequence of Borrelia afzelii K78 and comparative genome analysis.

    Directory of Open Access Journals (Sweden)

    Wolfgang Schüler

    Full Text Available The main Borrelia species causing Lyme borreliosis in Europe and Asia are Borrelia afzelii, B. garinii, B. burgdorferi and B. bavariensis. This is in contrast to the United States, where infections are exclusively caused by B. burgdorferi. Until to date the genome sequences of four B. afzelii strains, of which only two include the numerous plasmids, are available. In order to further assess the genetic diversity of B. afzelii, the most common species in Europe, responsible for the large variety of clinical manifestations of Lyme borreliosis, we have determined the full genome sequence of the B. afzelii strain K78, a clinical isolate from Austria. The K78 genome contains a linear chromosome (905,949 bp and 13 plasmids (8 linear and 5 circular together presenting 1,309 open reading frames of which 496 are located on plasmids. With the exception of lp28-8, all linear replicons in their full length including their telomeres have been sequenced. The comparison with the genomes of the four other B. afzelii strains, ACA-1, PKo, HLJ01 and Tom3107, as well as the one of B. burgdorferi strain B31, confirmed a high degree of conservation within the linear chromosome of B. afzelii, whereas plasmid encoded genes showed a much larger diversity. Since some plasmids present in B. burgdorferi are missing in the B. afzelii genomes, the corresponding virulence factors of B. burgdorferi are found in B. afzelii on other unrelated plasmids. In addition, we have identified a species specific region in the circular plasmid, cp26, which could be used for species determination. Different non-coding RNAs have been located on the B. afzelii K78 genome, which have not previously been annotated in any of the published Borrelia genomes.

  2. Low prevalence of Borrelia bavariensis in Ixodes ricinus ticks in southeastern Austria.

    Science.gov (United States)

    Glatz, Martin; Muellegger, Robert R; Hizo-Teufel, Cecilia; Fingerle, Volker

    2014-10-01

    Borrelia bavariensis was recently described as a distinct genospecies among the B. burgdorferi sensu lato complex. The prevalence of B. bavariensis in Austria, a highly endemic area for tick-transmitted pathogens, is scarcely characterized. To investigate the prevalence of B. bavariensis in Ixodes ricinus ticks we reevaluated the results of a study conducted in 518 ticks from southeastern Austria collected in 2002 and 2003. The presence of B. burgdorferi s.l.-specific DNA in ticks was analyzed by a PCR for the outer surface protein A (ospA) gene. Borrelia species were differentiated by restriction fragment length polymorphism (RFLP) analysis, and samples positive for B. bavariensis were further analyzed by multilocus sequence analysis. Two of 133 (1.5%) B. burgdorferi s.l.-positive I. ricinus ticks were infected with B. bavariensis. Both specimens were coinfected with the OspA serotype 5 of B. garinii. Borrelia bavariensis is present; however, seem to be rare in I. ricinus ticks in southeastern Austria.

  3. 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.

  4. 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.

  5. 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

  6. 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

  7. 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) ...

  8. Circumstantial evidence for an increase in the total number and activity of borrelia-infected ixodes ricinus in the Netherlands

    Directory of Open Access Journals (Sweden)

    Sprong Hein

    2012-12-01

    Full Text Available Abstract Background Between 1994 and 2009, a threefold increase has been observed in consultations of general practitioners for tick bites and Lyme disease in The Netherlands. The objective of this study was to determine whether an increase in the number of questing ticks infected with B. burgdorferi sensu lato is a potential cause of the rise in Lyme disease incidence. Methods Historic data on land usage, temperature and wildlife populations were collected and analyzed together with data from two longitudinal field studies on density of questing ticks. Effective population sizes of Borrelia burgdorferi s.l. were calculated. Results Long-term trend analyses indicated that the length of the annual tick questing season increased as well as the surface area of tick-suitable habitats in The Netherlands. The overall abundances of feeding and reproductive hosts also increased. Mathematical analysis of the data from the field studies demonstrated an increase in mean densities/activities of questing ticks, particularly of larvae between 2006 and 2009. No increase in infection rate of ticks with Borrelia burgdorferi sensu lato was found. Population genetic analysis of the collected Borrelia species points to an increase in B. afzelii and B. garinii populations. Conclusions Together, these findings indicate an increase in the total number of Borrelia-infected ticks, providing circumstantial evidence for an increase in the risk of acquiring a bite of a tick infected with B. burgdorferi s.l. Due to the high spatiotemporal variation of tick densities/activities, long-term longitudinal studies on population dynamics of I. ricinus are necessary to observe significant trends.

  9. NO ASSOCIATION BETWEEN tHbmass AND POLYMORPHISMS IN THE HBB GENE IN ENDURANCE ATHLETES.

    Science.gov (United States)

    Malczewska-Lenczowska, J; Orysiak, J; Majorczyk, E; Pokrywka, A; Kaczmarski, J; Szygula, Z; Sitkowski, D

    2014-06-01

    The aim of this study was to examine the association between tHbmass and HBB gene polymorphisms in athletes of endurance disciplines. Eighty-two well-trained athletes (female n=36, male n=46), aged 19.3 ± 2.7 years, representing cross country skiing (n=37) and middle- and long-distance running (n=45), participated in the study. Genotyping for 2 polymorphisms in the HBB gene (- 551C/T and intron 2, +16 C/G) was performed using restriction fragment length polymorphism analysis. Total haemoglobin mass (tHbmass) was determined by the optimized carbon monoxide rebreathing method. Blood morphology, indices of iron status (ferritin, transferrin receptor and total iron binding capacity) and C reactive protein were also determined. No differences were found in the HBB genotype and allele frequencies between male and female athletes. Regardless of the polymorphisms, no relationships were found between HBB genotypes as well as alleles and relative values of tHbmass, expressed per body mass (g · kg(-1) BM), both in female and male athletes. Our results demonstrated that -551 C/T and intron 2, +16 C/G polymorphisms of the HBB gene have no association with total haemoglobin mass in endurance athletes. It cannot be ruled out that several polymorphisms, each with a small but significant contribution, may be responsible for the amount of haemoglobin.

  10. 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.

  11. Improved serodiagnosis of early Lyme borreliosis: immunoblot with local Borrelia afzelii strain.

    Science.gov (United States)

    Jovicić, Vilma Lj; Grego, Edita M; Lako, Branislav L; Ristović, Blagoje M; Lepsanović, Zorica A; Stajković, Novica T

    2003-11-01

    To improve the serodiagnosis of Lyme borreliosis (LB) the performances of four tests were evaluated. An indirect immunofluorescent assay based on Borrelia burgdorferi s.s., enzyme-linked immunosorbent assay (ELISA) based on local isolates of Borrelia afzelii and B. burgdorferi s.s., and immunoblot (IB) of B. afzelii were prepared. The serum panels contained 214 serum samples: control group (n=120) and patients at different stages of LB (n=94). The specificity of IB was 96%, of in-house ELISA 93%, and of IFA 89%. In early LB the sensitivity of IFA was 36%, ELISA 67%, and IB 93%. In late-stage LB the sensitivity was: 72% for IFA, 80% for ELISA, and 94% for IB. Comparison of in-house and Behring ELISA showed that the sensitivity of the serological assay could be increased when the test was based on local Borrelia strains. IgM and IgG antibodies from sera of patients with early and late LB most frequently demonstrated reactivity to OspC. The other significant proteins in early LB were: p39, p41 in IgM IB, and p83/100, p39, Osp17 in IgG IB; in late LB: p39, p41 in IgM IB, and p83/100, Osp17, p21 and p43 in IgG IB. Using IB based on local B. afzelii isolates improves the serodiagnosis of early LB in our geographical region.

  12. 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.

  13. Constraining wrong-sign $hbb$ couplings with $h \\rightarrow \\Upsilon \\gamma$

    CERN Document Server

    Modak, Tanmoy; Sadhukhan, Soumya; Silva, João P; Srivastava, Rahul

    2016-01-01

    The rare decay $h \\rightarrow \\Upsilon \\gamma$ has a very small rate in the Standard Model, due to a strong cancellation between the direct and indirect diagrams. Models with a changed $hbb$ coupling can thus lead to a great increase in this decay. Current limits on two Higgs doublet models still allow for the possibility that the $hbb$ coupling might have a sign opposite to the Standard Model; the so-called "wrong-sign". We show how $h \\rightarrow \\Upsilon \\gamma$ can be used to put limits on the wrong-sign solutions.

  14. 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

  15. Comparative genome analysis: selection pressure on the Borrelia vls cassettes is essential for infectivity

    Directory of Open Access Journals (Sweden)

    Wilske Bettina

    2006-08-01

    Full Text Available Abstract Background At least three species of Borrelia burgdorferi sensu lato (Bbsl cause tick-borne Lyme disease. Previous work including the genome analysis of B. burgdorferi B31 and B. garinii PBi suggested a highly variable plasmid part. The frequent occurrence of duplicated sequence stretches, the observed plasmid redundancy, as well as the mainly unknown function and variability of plasmid encoded genes rendered the relationships between plasmids within and between species largely unresolvable. Results To gain further insight into Borreliae genome properties we completed the plasmid sequences of B. garinii PBi, added the genome of a further species, B. afzelii PKo, to our analysis, and compared for both species the genomes of pathogenic and apathogenic strains. The core of all Bbsl genomes consists of the chromosome and two plasmids collinear between all species. We also found additional groups of plasmids, which share large parts of their sequences. This makes it very likely that these plasmids are relatively stable and share common ancestors before the diversification of Borrelia species. The analysis of the differences between B. garinii PBi and B. afzelii PKo genomes of low and high passages revealed that the loss of infectivity is accompanied in both species by a loss of similar genetic material. Whereas B. garinii PBi suffered only from the break-off of a plasmid end, B. afzelii PKo lost more material, probably an entire plasmid. In both cases the vls gene locus encoding for variable surface proteins is affected. Conclusion The complete genome sequences of a B. garinii and a B. afzelii strain facilitate further comparative studies within the genus Borrellia. Our study shows that loss of infectivity can be traced back to only one single event in B. garinii PBi: the loss of the vls cassettes possibly due to error prone gene conversion. Similar albeit extended losses in B. afzelii PKo support the hypothesis that infectivity of Borrelia

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. HbbTV:HTML5将会成为电视的未来

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    HTML5是一个热议的焦点,当它被公认为手机未来的时候,也代表着电视的未来。11月18—19日在上海举行的2013亚太OTTTV峰会上,HbbTV亚太区总代表王哗先生从HTML5在业界的最新进展、HTML5对于OTT意味着什么、HTML5为PayTV运营商所青睐、HbbTV2.0语境中的HTML5、HTML5未来趋势等五个方面阐述了HTML5将会成为电视的未来。

  1. 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.

  2. 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

  3. 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.

  4. 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

  5. Characterization of the HBB: c.*233G > C Variant: No Evidence of a β-Thalassemic Phenotype.

    Science.gov (United States)

    Smith, Debra L; Mitui, Midori; Park, Jason Y; Luu, Hung S; Timmons, Charles F

    2016-01-01

    β-Thalassemia (β-thal) results from homozygous or compound heterozygous inheritance of β-globin alleles that yield decreased or absent synthesis of the β chain. Disease is frequently severe, requiring lifelong transfusion therapy. Heterozygosity for a β-thal allele results in an asymptomatic carrier state with mild but characteristic hematological findings. More than 200 β-globin alleles have been demonstrated to produce β-thal. For populations with a high prevalence of β-thal, screening for carrier status, genetic counseling and prenatal diagnosis are important components of efforts to both reduce disease incidence and provide early diagnosis and treatment. It is therefore important to define and characterize potential β-thal alleles. We sought to further characterize the previously reported β-thal allele, HBB: c.*233G > C. This variant is provisionally included in the HbVar database based on a study of Palestinians in the Gaza Strip with β-thal disease or carrier status (known or suspected) where 4.2% of subjects were found to have HBB: c.*233G > C. In our patient population, we detected the HBB: c.*233G > C variant in 17.3% of individuals (17 heterozygotes, one homozygote) undergoing β hemoglobin (Hb) gene sequencing at our laboratory over a 25-month period. Hematological parameters were analyzed to determine if these individuals demonstrated findings consistent with inheritance of a β-thal allele. Individuals with the HBB: c.*233G > C variant did not demonstrate any abnormalities in hematological parameters characteristic of β-thal carrier state (17 heterozygotes) or clinical evidence of disease (homozygote). Our data demonstrate no evidence for pathogenicity of the HBB: c.*233G > C variant but rather demonstrate that this variant is a common benign polymorphism.

  6. High Frequency of Hb E-Saskatoon (HBB: c.67G > A) in Brazilians: A New Genetic Origin?

    Science.gov (United States)

    Wagner, Sandrine C; Lindenau, Juliana D; Castro, Simone M de; Santin, Ana Paula; Zaleski, Carina F; Azevedo, Laura A; Ribeiro Dos Santos, Ândrea K C; Dos Santos, Sidney E B; Hutz, Mara H

    2016-08-01

    Hb E-Saskatoon [β22(B4)Glu→Lys, HBB: c.67G > A] is a rare, nonpathological β-globin variant that was first described in a Canadian woman of Scottish and Dutch ancestry and has since then been detected in several populations. The aim of the present study was to identify the origin of Hb E-Saskatoon in Brazil using β-globin haplotypes and genetic ancestry in carriers of this hemoglobin (Hb) variant. Blood samples were investigated by isoelectric focusing (IEF) and high performance liquid chromatography (HPLC) using commercial kits. Hb E-Saskatoon was confirmed by amplification of the HBB gene, followed by sequence analysis. Haplotypes of the β-globin gene were determined by polymerase chain reaction (PCR), followed by digestion with specific restriction enzymes. Individual ancestry was estimated with 48 biallelic insertion/deletions using three 16-plex PCR amplifications. The IEF pattern was similar to Hbs C (HBB: c.19G > A) and Hb E (HBB: c.79G > A) [isoelectric point (pI): 7.59-7.65], and HPLC results showed an elution in the Hb S (HBB: c.20A > T) window [retention time (RT): 4.26-4.38]. DNA sequencing of the amplified β-globin gene showed a mutation at codon 22 (GAA>AAA) corresponding to Hb E-Saskatoon. A total of 11 cases of this variant were identified. In nine unrelated individuals, Hb E-Saskatoon was in linkage disequilibrium with haplotype 2 [+ - - - -]. All subjects showed a high degree of European contribution (mean = 0.85). Hb E-Saskatoon occurred on the β-globin gene of haplotype 2 in all Brazilian carriers. These findings suggest a different genetic origin for this Hb variant from that previously described.

  7. 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.

  8. Future e(+)e(-) colliders' sensitivity to Hbb coupling and CP violation.

    Science.gov (United States)

    Braguta, V; Chalov, A; Likhoded, A; Rosenfeld, R

    2003-06-20

    We perform a complete simulation of the process e(+)e(-)-->bbvv, where nu can be an electron, muon, or tau neutrino, in the context of a general Higgs coupling to b quarks. We parametrize the Hbb; coupling as (m(b)/v)(a+igamma(5)b). Taking into account interference effects between pure Higgs and Standard Model contributions, we find that sensitivities of the order of 2% and 20% can be obtained at a future e(+)e(-) collider for deviations of the a and b parameters, respectively, from their Standard Model values. Combining our analysis with an independent measurement of Gamma(H-->bb) can provide evidence about the CP nature of the Higgs sector.

  9. 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

  10. Ixodes ricinus as vector and reservoir of Borrelia burgdorferi in an urban environment

    Directory of Open Access Journals (Sweden)

    Rajković Dragana V.

    2005-01-01

    Full Text Available As specific arthropods, ticks in urban environments transmit the agents of dangerous contagious diseases endangering human and animal health. They are vectors of numerous infective agents that cause serious diseases of animals such as babesiosis and ehrlichiosis. Tick populations maintain and transmit an increas­ing number of different infective agents. In regard to the human population, they transmit the agents of tick encephalitis, Omsk hemorrhagic disease, Crim-Congo hemorrhagic disease, Kyasa-nur forest fever, ixodid fever, Q fever, Rocky Mountain spotted fever, tularemia, Siberian tick typhus and Lyme disease the last of which according to acarological references has become the dominant zoonosis in over 140 of the world's countries. In urban environments ticks represent problem both for their epidemiological significance and as organisms whose presence disturbs people during their work and rest. In Europe, the two most fre­quent species and Ixodes ricinus and Ixodes persulcatus. Both species have a wide spectrum of hosts.

  11. 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

  12. Generation of KCL035 research grade human embryonic stem cell line carrying a mutation in HBB gene

    Directory of Open Access Journals (Sweden)

    Heema Hewitson

    2016-03-01

    Full Text Available The KCL035 human embryonic stem cell line was derived from an embryo donated for research that carried a mutation in the HBB gene, which is linked to the β-thalassemia syndrome. The ICM was isolated using laser microsurgery and plated on γ-irradiated human foreskin fibroblasts. Both the derivation and cell line propagation were performed in an animal product-free environment. Pluripotent state and differentiation potential were confirmed by in vitro assays.

  13. 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

  14. β-Thalassemia major resulting from compound heterozygosity for HBB: c.92+2T>C [formerly known as IVS-I-2 (T>C)] and a novel β(0)-thalassemia frameshift mutation: HBB: c.209delG; p.Gly70Valfs*20.

    Science.gov (United States)

    Kluge, Michelle L; Hoyer, James D; Swanson, Kenneth C; Oliveira, Jennifer L

    2014-01-01

    A novel β(0)-thalassemia (β-thal) frameshift mutation, HBB: c.209delG; p.Gly70Valfs*20, is described in a 21-year-old African American female with β-thalassemia major (β-TM) due to compound heterozygosity for the β(0)-thal mutation HBB: c.92+2T>C [formerly known as IVS-I-2 (T>C)] and HBB: c.209delG. The combination of these mutations demonstrates a complete lack of β-globin chain synthesis, evidenced by the proband having no Hb A present.

  15. Integrative Analysis of CRISPR/Cas9 Target Sites in the Human HBB Gene

    Directory of Open Access Journals (Sweden)

    Yumei Luo

    2015-01-01

    Full Text Available Recently, the clustered regularly interspaced short palindromic repeats (CRISPR system has emerged as a powerful customizable artificial nuclease to facilitate precise genetic correction for tissue regeneration and isogenic disease modeling. However, previous studies reported substantial off-target activities of CRISPR system in human cells, and the enormous putative off-target sites are labor-intensive to be validated experimentally, thus motivating bioinformatics methods for rational design of CRISPR system and prediction of its potential off-target effects. Here, we describe an integrative analytical process to identify specific CRISPR target sites in the human β-globin gene (HBB and predict their off-target effects. Our method includes off-target analysis in both coding and noncoding regions, which was neglected by previous studies. It was found that the CRISPR target sites in the introns have fewer off-target sites in the coding regions than those in the exons. Remarkably, target sites containing certain transcriptional factor motif have enriched binding sites of relevant transcriptional factor in their off-target sets. We also found that the intron sites have fewer SNPs, which leads to less variation of CRISPR efficiency in different individuals during clinical applications. Our studies provide a standard analytical procedure to select specific CRISPR targets for genetic correction.

  16. Integrative Analysis of CRISPR/Cas9 Target Sites in the Human HBB Gene.

    Science.gov (United States)

    Luo, Yumei; Zhu, Detu; Zhang, Zhizhuo; Chen, Yaoyong; Sun, Xiaofang

    2015-01-01

    Recently, the clustered regularly interspaced short palindromic repeats (CRISPR) system has emerged as a powerful customizable artificial nuclease to facilitate precise genetic correction for tissue regeneration and isogenic disease modeling. However, previous studies reported substantial off-target activities of CRISPR system in human cells, and the enormous putative off-target sites are labor-intensive to be validated experimentally, thus motivating bioinformatics methods for rational design of CRISPR system and prediction of its potential off-target effects. Here, we describe an integrative analytical process to identify specific CRISPR target sites in the human β-globin gene (HBB) and predict their off-target effects. Our method includes off-target analysis in both coding and noncoding regions, which was neglected by previous studies. It was found that the CRISPR target sites in the introns have fewer off-target sites in the coding regions than those in the exons. Remarkably, target sites containing certain transcriptional factor motif have enriched binding sites of relevant transcriptional factor in their off-target sets. We also found that the intron sites have fewer SNPs, which leads to less variation of CRISPR efficiency in different individuals during clinical applications. Our studies provide a standard analytical procedure to select specific CRISPR targets for genetic correction.

  17. 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...

  18. The effect of HBB:c.*+96T>C (3’UTR +1570 T>C on the mild b-thalassemia intermedia phenotype

    Directory of Open Access Journals (Sweden)

    Türker Bilgen

    2011-09-01

    Full Text Available Hemoglobin beta (HBB:c.*+96T>C substitution is very rare among β-globin gene mutations and its clinical significance remains to be clarified. The present study aimed to investigate the role of HBB:c.*+96T>C in the β-thalassemia intermedia phenotype in a Turkish family. The proband and parents were screened for β-globin gene mutations via direct sequencing. Hematological and physical examination results were recorded, and correlated according to genotype. The proband was compound heterozygous for Cod 8 (-AA and HBB:c.*+96T>C, whereas his mother and father were heterozygous for Cod 8 (-AA and HBB:c.*+96T>C, respectively. The father had almost normal hematological findings, whereas the mother had the typical β-thalassemia trait phenotype. The proband was diagnosed as mild β-thalassemia intermedia based on hepatosplenomegaly and hematological findings. To the best of our knowledge this is the first report of HBB:c.*+96T>C mutation in a Turkish family. HBB:c.* 96T>C substitution is a very rare, but clinically relevant β-globin gene mutation. Additionally, we think that if 1 spouse is a carrier for β-globin gene mutation the other should be screened for silent mutations, such as HBB:c.*+96T>C mutation of the β-globin gene, even if she/he does not have any clinical or hematological signs of the β-thalassemia trait phenotype.

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. Chronic inflammatory state in sickle cell anemia patients is associated with HBB(*)S haplotype.

    Science.gov (United States)

    Bandeira, Izabel C J; Rocha, Lillianne B S; Barbosa, Maritza C; Elias, Darcielle B D; Querioz, José A N; Freitas, Max Vitor Carioca; Gonçalves, Romélia P

    2014-02-01

    The chronic inflammatory state in sickle cell anemia (SCA) is associated with several factors such as the following: endothelial damage; increased production of reactive oxygen species; hemolysis; increased expression of adhesion molecules by leukocytes, erythrocytes, and platelets; and increased production of proinflammatory cytokines. Genetic characteristics affecting the clinical severity of SCA include variations in the hemoglobin F (HbF) level, coexistence of alpha-thalassemia, and the haplotype associated with the HbS gene. The different haplotypes of SCA are Bantu, Benin, Senegal, Cameroon, and Arab-Indian. These haplotypes are associated with ethnic groups and also based on the geographical origin. Studies have shown that the Bantu haplotype is associated with higher incidence of clinical complications than the other haplotypes and is therefore considered to have the worst prognosis. This study aimed to evaluate the profile of the proinflammatory cytokines interleukin-6, tumor necrosis factor-α, and interleukin-17 in patients with SCA and also to assess the haplotypes associated with beta globin cluster S (HBB(*)S). We analyzed a total of 62 patients who had SCA and had been treated with hydroxyurea; they had received a dose ranging between 15 and 25 (20.0±0.6)mg/kg/day for 6-60 (18±3.4)months; their data were compared with those for 30 normal individuals. The presence of HbS was detected and the haplotypes of the beta S gene cluster were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our study demonstrated that SCA patients have increased inflammatory profile when compared to the healthy individuals. Further, analysis of the association between the haplotypes and inflammatory profile showed that the levels of IL-6 and TNF-α were greater in subjects with the Bantu/Bantu haplotype than in subjects with the Benin/Benin haplotype. The Bantu/Benin haplotype individuals had lower levels of cytokines than those with

  4. Oligoarthritis caused by Borrelia bavariensis, Austria, 2014.

    Science.gov (United States)

    Markowicz, Mateusz; Ladstatter, Stefan; Schotta, Anna M; Reiter, Michael; Pomberger, Gerhard; Stanek, Gerold

    2015-06-01

    A case of Lyme oligoarthritis occurred in an 11-year-old boy in Vienna, Austria. DNA of Borrelia bavariensis was detected by PCR in 2 aspirates obtained from different joints. Complete recovery was achieved after a 4-week course with amoxicillin. Lyme arthritis must be considered in patients from Europe who have persisting joint effusions.

  5. FP7 HBB-Next, a proposal for a European 7th Framework Project on next-generation Hybrid Broadcast Broadband

    NARCIS (Netherlands)

    Deventer, M.O. van; Niamut, O.A.

    2011-01-01

    This is the FP7 proposal HBB-Next: Next-Generation Hybrid Broadcast Broadband. It was submitted on 18 January 2011 for FP7 call no.7, objective 1.5a. So far, Hybrid Broadcast Internet applications have only linked Internet and broadcast content by offering over‐the‐top On‐Demand content in addition

  6. Are Apodemus spp. mice and Myodes glareolus reservoirs for Borrelia miyamotoi, Candidatus Neoehrlichia mikurensis, Rickettsia helvetica, R. monacensis and Anaplasma phagocytophilum?

    Science.gov (United States)

    Burri, C; Schumann, O; Schumann, C; Gern, L

    2014-04-01

    In Europe, in addition to Borrelia burgdorferi sensu lato and tick-borne encephalitis (TBE) virus, other zoonotic pathogens, like B. miyamotoi, a species related to the relapsing fever spirochaetes, Candidatus Neoehrlichia mikurensis (N. mikurensis), Rickettsia helvetica, Rickettsia monacensis, and Anaplasma phagocytophilum have been reported in the ixodid tick Ixodes ricinus. No study was conducted to identify reservoir hosts for these pathogens. Here, we investigated the role played by wild rodents in the natural transmission cycle of B. miyamotoi, N. mikurensis, R. helvetica, R. monacensis, and A. phagocytophilum in Switzerland. In 2011 and 2012, small mammals were captured in an area where these pathogens occur in questing ticks. Ixodes ricinus ticks infesting captured small mammals were analysed after their moult by PCR followed by reverse line blot to detect the different pathogens. Xenodiagnostic larvae were used to evaluate the role of rodents as reservoirs and analysed after their moult. Most of the 108 captured rodents (95.4%) were infested by I. ricinus ticks; 4.9%, 3.9%, 24.0%, and 0% of the rodents were infested by Borrelia, N. mikurensis, Rickettsia spp., and A. phagocytophilum-infected larvae, respectively. Borrelia afzelii, B. miyamotoi, N. mikurensis, Rickettsia spp., and A. phagocytophilum were detected in 2.8%, 0.17%, 2.6%, 6.8%, and 0% of the ticks attached to rodents, respectively. Borrelia afzelii was transmitted by 4 rodents to 41.2% of the xenodiagnostic ticks, B. miyamotoi by 3 rodents to 23.8%, and N. mikurensis was transmitted by 6 rodents to 41.0% of the xenodiagnostic ticks. None of the tested rodent transmitted Rickettsia spp. or A. phagocytophilum to I. ricinus xenodiagnostic larvae. This study showed that rodents are reservoir hosts for B. miyamotoi and N. mikurensis in Europe. PMID:24582511

  7. Identification of a Novel β-Globin Mutation (HBB: C.189_195delTCATGGC) in a Chinese Family.

    Science.gov (United States)

    He, Sheng; Lin, Li; Wei, Yuan; Chen, Biyan; Yi, Shang; Chen, Qiuli; Qiu, XiaoXia; Wei, Hongwei; Li, Guojian; Zheng, Chenguang

    2016-08-01

    β-Thalassemia (β-thal) is one of the most common genetic disorders worldwide. Molecular characterization of β-thal is essential for prevention and understanding the biology of the disease. More and more rare and novel mutations are being reported. Here, we report a novel 7 bp deletion at codons 63-65 (HBB: c.189_195delTCATGGC) in exon 2 of the β-globin gene in a family from Guangxi Province, China. This novel mutation causes a shift in the normal reading frame of the β-globin coding sequence and created a stop codon at codon 87 in exon 2, which leads to a β(0)-thal phenotype. PMID:27492766

  8. Exploitation of complement regulatory proteins by Borrelia and Francisella.

    Science.gov (United States)

    Madar, Marian; Bencurova, Elena; Mlynarcik, Patrik; Almeida, André M; Soares, Renata; Bhide, Katarina; Pulzova, Lucia; Kovac, Andrej; Coelho, Ana V; Bhide, Mangesh

    2015-06-01

    Pathogens have developed sophisticated mechanisms of complement evasion such as binding to the host complement regulatory proteins (CRPs) on their surface or expression of CRP mimicking molecules. The ability of pathogens to evade the complement system has been correlated with pathogenesis and host selectivity. Hitherto, little work has been undertaken to determine whether Borrelia and Francisella exploit various CRPs to block complement attack. Seventeen Borrelia (twelve species) and six Francisella (three subspecies) strains were used to assess their ability to bind human, sheep and cattle CRPs or mimic membrane associated complement regulators. A series of experiments including affinity ligand binding experiments, pull-down assays and mass spectrometry based protein identification, revealed an array of CRP binding proteins of Borrelia and Francisella. Unlike Francisella, Borrelia strains were able to bind multiple human CRPs. Three strains of Borrelia (SKT-4, SKT-2 and HO14) showed the presence of a human CD46-homologous motif, indicating their ability to possess putative human CD46 mimicking molecules. Similarly, five strains of Borrelia and two strains of Francisella may have surface proteins with human CD59-homologous motifs. Among ovine and bovine CRPs, the only CRP bound by Francisella (LVS, Tul4 strain) was vitronectin, while ovine C4BP, ovine factor H and bovine factor H were bound to Borrelia strains SKT-2, DN127 and Co53. This study presents an array of proteins of Borrelia and Francisella that bind CRPs or may mimic membrane-CRPs, thus enabling multiphasic complement evasion strategies of these pathogens.

  9. Multispacer sequence typing relapsing fever Borreliae in Africa.

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    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.

  10. 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...

  11. Immune response to Lactobacillus plantarum expressing Borrelia burgdorferi OspA is modulated by the lipid modification of the antigen.

    Directory of Open Access Journals (Sweden)

    Beatriz del Rio

    Full Text Available BACKGROUND: Over the past decade there has been increasing interest in the use of lactic acid bacteria as mucosal delivery vehicles for vaccine antigens, microbicides and therapeutics. We investigated the mechanism by which a mucosal vaccine based in recombinant lactic acid bacteria breaks the immunological tolerance of the gut in order to elicit a protective immune response. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed how the lipid modification of OspA affects the localization of the antigen in our delivery vehicle using a number of biochemistry techniques. Furthermore, we examined how OspA-expressing L. plantarum breaks the oral tolerance of the gut by stimulating human intestinal epithelial cells, peripheral blood mononuclear cells and monocyte derived dendritic cells and measuring cytokine production. We show that the leader peptide of OspA targets the protein to the cell envelope of L. plantarum, and it is responsible for protein export across the membrane. Mutation of the lipidation site in OspA redirects protein localization within the cell envelope. Further, we show that lipidated-OspA-expressing L. plantarum does not induce secretion of the pro-inflammatory cytokine IL-8 by intestinal epithelial cells. In addition, it breaks oral tolerance of the gut via Th1/Th2 cell mediated immunity, as shown by the production of pro- and anti-inflammatory cytokines by human dendritic cells, and by the production of IgG2a and IgG1 antibodies, respectively. CONCLUSIONS/SIGNIFICANCE: Lipid modification of OspA expressed in L. plantarum modulates the immune response to this antigen through a Th1/Th2 immune response.

  12. DNA damage in leukocytes of sickle cell anemia patients is associated with hydroxyurea therapy and with HBB*S haplotype.

    Science.gov (United States)

    da Silva Rocha, Lilianne Brito; Dias Elias, Darcielle Bruna; Barbosa, Maritza Cavalcante; Bandeira, Izabel Cristina Justino; Gonçalves, Romélia Pinheiro

    2012-12-12

    Hydroxyurea (HU) is the primary pharmacologic agent for preventing the complications and improving the quality of life of sickle cell anemia (SCA) patients. Although HU has been associated with an increased risk of leukemia in some patients with myeloproliferative disorders, the mutagenic and carcinogenic potential of HU has not been established. This study used the alkaline comet assay to investigate DNA damage in peripheral blood leukocytes from 41 individuals with SCA treated with HU (SCAHU) and from 26 normal individuals. The presence of HbS and the analysis of the haplotypes of the beta S gene cluster were done by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The damage index (DI) in the SCAHU group was significantly higher than in controls (p20kg/m(2). No significant influence of mean HU dose was observed on DI (p=0.950). However, individuals who received a mean HU dose≥20mg/kg showed a higher DI than those who received less. Furthermore, an association was observed between DI damage and HBB*S gene haplotypes. DI values for the Bantu/Bantu haplotype was greater when compared to the Benin/Benin haplotype; and the Bantu/Benin haplotype had a DI lower than the Bantu/Bantu haplotype and greater than the Benin/Benin haplotype. Our results show that DNA damage in sickle cell anemia is associated not only with treatment with HU but also with genotype. PMID:22918118

  13. Seroprevalencia de la infección por Borrelia burgdorgferi y Rickettsia conorii en población humana y canina de la zona básica de salud de San Andrés del Rabanedo (León, España

    Directory of Open Access Journals (Sweden)

    Rojo Vázquez Jaime

    1997-01-01

    Full Text Available FUNDAMENTOS: Se estudia la seroprevalencia de la infección por Borrelia burgdorferi y Rickettsia conorii en población humana y canina para conocer la situación de ambas en humanos, al mismo tiempo que la significación del perro, como indicador de la circulación de estos agentes entre aquéllos, en la Zona de Salud de San Andrés del Rabanedo, León. MÉTODO: Se realizó un estudio en 98 sueros humanos y 95 caninos (de diversas razas y aptitudes frente a B. burgdorferi (títulos de positividad > a 1/128 y > a 1/64 o superiores, respectivamente y 104 sueros humanos y 84 caninos frente a R. conorii (positividad a título > a 1/64 o superiores en ambas especies mediante inmunofluorescencia indirecta (IFI. RESULTADOS: Se halló positividad a las dos infecciones, tanto en personas como en perros. Frente a B. burgdorferi fue superior en humanos que en perros y frente a R. conorii fue superior en éstos que en humanos. En personas fue del 4,08% frente a B. burgdorferi y 1% frente a R. conorii; en perros fue del 2,10% frente a B. burgdorferi y del 14,28% frente a R. conorii. Los valores más altos se hallaron en los meses de primavera-verano, salvo en el caso de B. burgdorferi en perros. La seroprevalencia fue mayor en perros dedicados al cuidado de ganado (ovino que en los de caza y guarda. CONCLUSIONES: Los porcentajes de seroprevalencia hallados en nuestro trabajo, tanto en seres humanos como en caninos, considerados en el ámbito territorial de una zona geográfica semi-rural de la provincia de León, han sido iguales o inferiores a los reseñados para otras provincias, incluida la totalidad de la de León. En perros se halló mayor seroprevalencia frente a R. conorii que frente a B. burgdorferi, lo que indica que es el agente más extendido en nuestra Provincia, como han señalado otros autores. Los valores hallados en seres humanos frente a B. burgdorferi han sido más altos que en perros; la existencia de reacciones cruzadas con otros

  14. 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

  15. Transstadial transmission of Borrelia turcica in Hyalomma aegyptium ticks.

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    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.

  16. 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 - ...

  17. The genetic basis of asymptomatic codon 8 frame-shift (HBB:c25_26delAA) β(0) -thalassaemia homozygotes.

    Science.gov (United States)

    Jiang, Zhihua; Luo, Hong-Yuan; Huang, Shengwen; Farrell, John J; Davis, Lance; Théberge, Roger; Benson, Katherine A; Riolueang, Suchada; Viprakasit, Vip; Al-Allawi, Nasir A S; Ünal, Sule; Gümrük, Fatma; Akar, Nejat; Başak, A Nazli; Osorio, Leonor; Badens, Catherine; Pissard, Serge; Joly, Philippe; Campbell, Andrew D; Gallagher, Patrick G; Steinberg, Martin H; Forget, Bernard G; Chui, David H K

    2016-03-01

    Two 21-year old dizygotic twin men of Iraqi descent were homozygous for HBB codon 8, deletion of two nucleotides (-AA) frame-shift β(0) -thalassaemia mutation (FSC8; HBB:c25_26delAA). Both were clinically well, had splenomegaly, and were never transfused. They had mild microcytic anaemia (Hb 120-130 g/l) and 98% of their haemoglobin was fetal haemoglobin (HbF). Both were carriers of Hph α-thalassaemia mutation. On the three major HbF quantitative trait loci (QTL), the twins were homozygous for G>A HBG2 Xmn1 site at single nucleotide polymorphism (SNP) rs7482144, homozygous for 3-bp deletion HBS1L-MYB intergenic polymorphism (HMIP) at rs66650371, and heterozygous for the A>C BCL11A intron 2 polymorphism at rs766432. These findings were compared with those found in 22 other FSC8 homozygote patients: four presented with thalassaemia intermedia phenotype, and 18 were transfusion dependent. The inheritance of homozygosity for HMIP 3-bp deletion at rs66650371 and heterozygosity for Hph α-thalassaemia mutation was found in the twins and not found in any of the other 22 patients. Further studies are needed to uncover likely additional genetic variants that could contribute to the exceptionally high HbF levels and mild phenotype in these twins. PMID:26771086

  18. 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.

  19. Prevalence of avian influenza viruses, Borrelia garinii, Mycobacterium avium, and Mycobacterium avium subsp. paratuberculosis in waterfowl and terrestrial birds in Slovakia, 2006.

    Science.gov (United States)

    Gronesova, Paulina; Ficova, Martina; Mizakova, Adriana; Kabat, Peter; Trnka, Alfred; Betakova, Tatiana

    2008-10-01

    The prevalence of Borrelia, Mycobacteria and avian influenza virus (AIV) infections, together with the distribution of different AIV subtypes, was studied in migratory waterfowl and terrestrial birds trapped in three localities in Slovakia during 2006. Samples obtained from waterfowl captured in the Senianske Ponds area of Eastern Slovakia showed the highest diversity of AIV isolates. A total of 13 different subtypes were detected in 19 samples from this location (H1N2, H2N2, H3N2, H6N6, H7N6, H9N2, H9N5, H9N6, H10N5, H10N6, H12N6, H13N6, and H16N6). H3N5 virus was detected in 50% of passerines testing positive for AIV in the Parizske Wetlands, with H7N2, H9N2, H9N5, H12N1, and H13N2 infections also recorded at this locality. H9N5 virus predominated in passerines captured at Trnava Ponds, with isolates H1N6, H6N5, H7N2, H7N6, H10N3, and H10N6 also detected at this location. There were five cases where different AIV infections were detected in oropharyngeal and cloacal samples originating from the same bird (H13N6 and H1N2; H10N5 and H12N6; H9N5 and H6N5; H10N6 and H7N6; and H9N2 and H3N5 in the oropharynx and cloaca, respectively). Between 21% and 52% of captured birds tested positive for Borrelia burgdorferi sensu lato, with the proportion infected depending on bird species and locality. Samples were characterized by polymerase chain reaction-restriction fragment length polymorphism analysis and identified as Borrelia garinii species (either B/B' or R/R' pattern). Mycobacteria were detected in 42% and 26% of waders captured at Senianske Ponds and marsh-dwelling passerines captured in the Parizske Wetlands, respectively. Interestingly, forest-dwelling passerine species caught in the Trnava Ponds region were tested negative for Mycobacteria. PMID:18798030

  20. 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.

  1. [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

  2. 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

  3. 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 ...

  4. Persistence of the antibody response to the VlsE sixth invariant region (IR6) peptide of Borrelia burgdorferi after successful antibiotic treatment of Lyme disease.

    Science.gov (United States)

    Peltomaa, Miikka; McHugh, Gail; Steere, Allen C

    2003-04-15

    It has been suggested that a Lyme disease. We studied the response to this peptide in 77 patients with early or late disease, for whom archival samples were available at the time of antibiotic treatment and approximately 6 months or years later. Eight (33%) of the 24 patients with early manifestations and 18 (86%) of the 21 patients with late manifestations had a Lyme disease.

  5. 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.

  6. 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

  7. 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.

  8. Which forest bird species are the main hosts of the tick, Ixodes ricinus, the vector of Borrelia burgdorferi sensu lato, during the breeding season?

    Science.gov (United States)

    Marsot, M; Henry, P-Y; Vourc'h, G; Gasqui, P; Ferquel, E; Laignel, J; Grysan, M; Chapuis, J-L

    2012-07-01

    Wild birds are important hosts for vector-borne pathogens, especially those borne by ticks. However, few studies have been conducted on the role of different bird species within a community as hosts of vector-borne pathogens. This study addressed individual and species factors that could explain the burden of Ixodes ricinus on forest birds during the reproductive periods of both vectors and hosts. The goal was to identify which bird species contribute the most to the tick population at the community level. Birds were mist-netted on four plots in 2008 and on seven plots in 2009 in two forests (Sénart and Notre Dame, near Paris, France). The dependence of the tick load per bird upon environmental conditions (questing nymph density, year and plot) and on host species traits (species, age, sex, body size, vertical space use, level of innate and acquired immunity) was analysed. Finally, the relative contribution of each bird species to the local dynamics of ticks was estimated, while accounting for their respective abundance. Tick burden differed markedly between bird species and varied according to questing nymph density. Bird species with a high body mass, those that forage low in the vegetation, and those that had a high innate immune response and a high spleen mass were more likely to have a high tick burden. Four species (the Common Blackbird, Turdus merula, the European Robin, Erithacus rubecula, the Song Thrush, Turdus philomelos, and the Winter Wren, Troglodytes troglodytes) hosted more than 90% of the ticks in the local bird community. These species, and particularly T. merula which was host to a high proportion of the nymphs, are likely to contribute significantly to the circulation of pathogens for which they are competent, such as the agent of Lyme borreliosis.

  9. 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.

  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. High-prevalence Borrelia miyamotoi infection among [corrected] wild turkeys (Meleagris gallopavo) in Tennessee.

    Science.gov (United States)

    Scott, M C; Rosen, M E; Hamer, S A; Baker, E; Edwards, H; Crowder, C; Tsao, J I; Hickling, G J

    2010-11-01

    During spring and fall 2009, 60 wild turkeys (Meleagris gallopavo) harvested by Tennessee hunters were surveyed for Borrelia spp. by sampling their blood, tissue, and attached ticks. In both seasons, 70% of turkeys were infested with juvenile Amblyomma americanum; one spring turkey hosted an adult female Ixodes brunneus. Polymerase chain reaction assays followed by DNA sequencing indicated that 58% of the turkeys were positive for the spirochete Borrelia miyamotoi, with tissue testing positive more frequently than blood (P = 0.015). Sequencing of the 16S-23S rRNA intergenic spacer indicated > or = 99% similarity to previously published sequences of the North American strain of this spirochete. Positive turkeys were present in both seasons and from all seven middle Tennessee counties sampled. No ticks from the turkeys tested positive for any Borrelia spp. This is the first report of B. miyamotoi in birds; the transmission pathways and epidemiological significance of this high-prevalence spirochetal infection remain uncertain.

  13. A new β(0) frameshift mutation, HBB: c.44delT (p.Leu14ArgfsX5), identified in an Argentinean family associated with secondary genetic modifiers of β-thalassemia.

    Science.gov (United States)

    Pepe, Carolina; Eberle, Silvia Eandi; Chaves, Alejandro; Milanesio, Berenice; Aguirre, Fernando M; Gómez, Vanesa Avalos; Diaz, Lilian; Mansini, Adrian P; Fernandez, Diego A; Sciuccati, Gabriela; Candas, Andrea; Cervio, Carolina; Bonduel, Mariana; Feliú-Torres, Aurora

    2014-01-01

    β-Thalassemia intermedia (β-TI) patients present with a wide spectrum of phenotypes depending on the presence of primary, secondary, and tertiary genetic modifiers which modulate, by different mechanisms, the degree of imbalance between α and β chains. Here we describe a new β(0) frameshift mutation, HBB: c.44delT (p.Leu14ArgfsX5), identified in four members of a family, associated with secondary genetic modifiers in three of them. The different genotype present in this family was suspected after hematological analysis and thorough observation of blood smears highlighting their importance in the identification of β-TI patients among members of the same family.

  14. No evidence for the diagnostic value of Borrelia serology in patients with sudden hearing loss.

    NARCIS (Netherlands)

    Bakker, R.; Aarts, M.C.; Heijden, G.J. van der; Rovers, M.M.

    2012-01-01

    In this evidence-based case report, we address the following clinical question: What is the predictive value of serological testing for Borrelia for diagnosing neuroborreliosis in patients with sudden sensorineural hearing loss? We searched for relevant articles in PubMed, Embase, and Web of Science

  15. A Novel Animal Model of Borrelia recurrentis Louse-Borne Relapsing Fever Borreliosis Using Immunodeficient Mice

    NARCIS (Netherlands)

    Larsson, C.; Lundqvist, J.; Rooijen, van N.; Bergstrom, S.

    2009-01-01

    Louse-borne relapsing fever (LBRF) borreliosis is caused by Borrelia recurrentis, and it is a deadly although treatable disease that is endemic in the Horn of Africa but has epidemic potential. Research on LBRF has been severely hampered because successful infection with B. recurrentis has been achi

  16. 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

  17. 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...

  18. 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 ...

  19. High Prevalence of Borrelia miyamotoi among Adult Blacklegged Ticks from White-Tailed Deer.

    Science.gov (United States)

    Han, Seungeun; Hickling, Graham J; Tsao, Jean I

    2016-02-01

    We compared the prevalence of Borrelia miyamotoi infection in questing and deer-associated adult Ixodes scapularis ticks in Wisconsin, USA. Prevalence among deer-associated ticks (4.5% overall, 7.1% in females) was significantly higher than among questing ticks (1.0% overall, 0.6% in females). Deer may be a sylvatic reservoir for this newly recognized zoonotic pathogen. PMID:26811985

  20. 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...

  1. β-Thalassemia Due to Intronic LINE-1 Insertion in the β-Globin Gene (HBB): Molecular Mechanisms Underlying Reduced Transcript Levels of the β-GlobinL1 Allele

    Science.gov (United States)

    Lanikova, Lucie; Kucerova, Jana; Indrak, Karel; Divoka, Martina; Issa, Jean-Pierre; Papayannopoulou, Thalia; Prchal, Josef T.; Divoky, Vladimir

    2016-01-01

    We describe the molecular etiology of β+-thalassemia that is caused by the insertion of the full-length transposable element LINE-1 (L1) into the intron-2 of the β-globin gene (HBB). The transcript level of the affected β-globin gene was severely reduced. The remaining transcripts consisted of full-length, correctly processed β-globin mRNA and a minute amount of three aberrantly spliced transcripts with a decreased half-life due to activation of the nonsense-mediated decay pathway. The lower steady-state amount of mRNA produced by the β-globinL1 allele also resulted from a reduced rate of transcription and decreased production of full-length β-globin primary transcripts. The promoter and enhancer sequences of the β-globinL1 allele were hypermethylated; however, treatment with a demethylating agent did not restore the impaired transcription. A histone deacetylase inhibitor partially reactivated the β-globinL1 transcription despite permanent β-globinL1 promoter CpG methylation. This result indicates that the decreased rate of transcription from the β-globinL1 allele is associated with an altered chromatin structure. Therefore, the molecular defect caused by intronic L1 insertion in the β-globin gene represents a novel etiology of β-thalassemia. PMID:23878091

  2. 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.

  3. 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...

  4.  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.

  5. "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.

  6. 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...

  7. 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.

  8. Fatal spirochetosis due to a relapsing fever-like Borrelia sp. in northern spotted owl

    Science.gov (United States)

    Thomas, N.J.; Bunikis, J.; Barbour, A.G.; Wolcott, M.J.

    2002-01-01

    Acute septicemic spirochetosis was diagnosed in an adult male northern spotted owl (Strix occidentalis caurina) found dead in Kittitas County, Washington, USA. Gross necropsy findings included marked enlargement of the liver and spleen and serofibrinous deposits on the serous membranes lining the body cavities and the pericardial and perihepatic sacs. Microscopic observations included macrophage infiltration in the liver and spleen with mild thrombosis and multifocal necrosis, as well as hemorrhage and acute inflammation in the choroid plexus of the brain. No viruses or pathogenic bacteria were isolated from brain, liver, or spleen, and no parasites were found in blood smears or impression smears of the liver. Chlamydial culture attempts were unsuccessful and no chlamydial antibodies were detected in serum. In silver-stained microscopic sections and by transmission electron microscopy of liver, numerous long, thin, spiral-shaped bacteria were seen in the liver, spleen, cerebral ventricles, and within blood vessels in many organs. The organism was identified as a member of the Borrelia genus by sequence analysis of the PCR-amplified 16S rRNA gene. The most closely related species is B. hermsii, an agent of relapsing fever in humans in the western United States. This is the first report of a relapsing fever-related Borrelia in a wild bird.

  9. 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

  10. 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.

  11. 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.

  12. 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.

  13. 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

    , the construction of a diagnostic score, evaluation of the scoring method using an independent dataset and an assessment of the analytical quality of the multiplex assay. The VlsE IgG had the highest diagnostic value with an AUC (area under the curve) of 96% on the receiver operating characteristic curve. The Osp......C IgM had AUCs just above 80%. All the other antigens had both low quantitative reactivity and lower contrast in the patients with LNB compared to controls. The diagnostic value of the assay may be improved by using a logistic model giving a sensitivity of 90 and 79% for the specificities at 92 and 98......%, respectively. Overall, the patients with LNB had serum reactivity in IgG VlsE, but modest antibody reactivity in the remaining 12 IgG and 13 IgM antibody measurements. Using a logistic regression model with five IgG and two IgM antigens, the sensitivity and specificity of the assay was improved; but the Ig...

  14. Borrelia sp. in naturally infected Didelphis aurita (Wied, 1826 (marsupialia: didelphidae

    Directory of Open Access Journals (Sweden)

    Isis dos Santos Abel

    2000-01-01

    Full Text Available Fifty-six opossums (Didelphis aurita were captured on the campus of Universidade Federal Rural do Rio de Janeiro, Seropédica county, Rio de Janeiro state, in order to investigate the occurrence of Borrelia sp among them in relation with the study of spirochaetemia and its ectoparasites. Blood tests were made through dark field and phase contrast microscopy, as well as the obtainment of blood smears. Smears were stained with Giemsa stain, which did not prove efficacy. There was no relation between results obtained through blood tests (13 opossum positive for Borreliasp., and this technique (two positive animals. Parasitaemia studies of 37 animals kept in captivity as well as of several recaptures in which animals once negative proved to be positive days later, showed that haemoscopical studies could be used as an effective diagnosis tool. Ectoparasites from nine animals were classified; with the occurrence of nymphal Amblyomma cajennense and adult Ctenocephalides sp..Cinquenta e seis gambás (Didelphis aurita foram capturados no campus da Universidade Federal Rural do Rio de Janeiro, Seropédica - Rio de Janeiro, com o intuito de se investigar a ocorrência de Borrelia sp. entre eles, associada ao estudo da espiroquetemia e de seus ectoparasitos. Amostras de sangue dos animais foram examinadas à microscopia de campo escuro e contraste de fase, além deste ter se destinado à obtenção de esfregaços sanguíneos, corados pelo método de Giemsa. Não houve relação entre os resultados obtidos através dos exames diretos do sangue (13 gambás positivos para Borrelia sp. e aqueles observados nos esfregaços sanguíneos (dois animais positivos, o que demonstra a baixa eficácia desta técnica. Estudos sobre a parasitemia de 37 animais mantidos em cativeiro, assim como as várias recapturas nas quais animais uma vez negativos mostraram-se positivos dias mais tarde, revelaram que os estudos hemoscópicos podem ser utilizados eficientemente como

  15. Tick-borne Relapsing Fever and Borrelia hermsii, Los Angeles County, California, USA

    Science.gov (United States)

    Raffel, Sandra J.; Schrumpf, Merry E.; Webster, Larry S.; Marques, Adriana R.; Spano, Robyn; Rood, Michael; Burns, Joe; Hu, Renjie

    2009-01-01

    The primary cause of tick-borne relapsing fever in western North America is Borrelia hermsii, a rodent-associated spirochete transmitted by the fast-feeding soft tick Ornithodoros hermsi. We describe a patient who had an illness consistent with relapsing fever after exposure in the mountains near Los Angeles, California, USA. The patient’s convalescent-phase serum was seropositive for B. hermsii but negative for several other vector-borne bacterial pathogens. Investigations at the exposure site showed the presence of O. hermsi ticks infected with B. hermsii and the presence of rodents that were seropositive for the spirochete. We determined that this tick-borne disease is endemic to the San Gabriel Mountains near the greater Los Angeles metropolitan area. PMID:19624916

  16. Borrelia theileri: observação em carrapatos do gênero Boophilus microplus no município de Guaíba, RS, Brasil Borrelia theileri: observation on Boophilus microplus ticks in Guaiba, RS, Brazil

    Directory of Open Access Journals (Sweden)

    João Ricardo Martins

    1996-12-01

    Full Text Available Espiroquetas da espécie Borrelia theileri identificadas em uma estirpe de carrapatos Boophilus microplus provenientes do município de Guaíba, RS. A observação ocorreu no exame de hemolinfa de fêmeas adultas com 10 dias pós-repleçâo, corada por Giemsa. Não foram observadas espiroquetas em ovos provenientes de teleóginas infectadas. A detecção da estirpe infectada sugere a presença de borreliose em rebanhos bovinos, fato que eventualmente pode interferir em resultados de diagnóstico ou tornar-se motivo de preocupação em produtos derivados de sangue bovino tais como vacinas vivas contra anaplasmose e babesiose bovina.Spirochetes of species Borrelia theileri were identifica in afield-strain of the caule tick Boophilus microplus, in Guaíba, RS, Brazil. Hemolymph smears from females 10 days post-repletion were collected by gentty section of the tarsal-metatarsaijoint, and dropped onto a microscope slide, and stained by Giemsa. No spirochetes were observed in eggs squashed and stained by Giemsa from the same infected strain. The detection of B. microplus adult females infected with Borrelia theileri suggesfs the likely presence of borreliosis in bovine heras what might eventually interfere with the interpretation of diagnosis results or become cause for concern in blood products such as anaplasmosis and babesiosis live vaccines.

  17. Insights into Borrelia miyamotoi infection from an untreated case demonstrating relapsing fever, monocytosis and a positive C6 Lyme serology.

    Science.gov (United States)

    Sudhindra, Praveen; Wang, Guiqing; Schriefer, Martin E; McKenna, Donna; Zhuge, Jian; Krause, Peter J; Marques, Adriana R; Wormser, Gary P

    2016-09-01

    We describe a patient from the United States with PCR- and serology-confirmed Borrelia miyamotoi infection who recovered without antibiotics. Our findings suggest that B. miyamotoi infection may cause relapsing fever, blood monocytosis and antibody reactivity to the C6 peptide. Further studies are required to better define the spectrum of clinical and laboratory findings for this emerging tick-transmitted infection. PMID:27412815

  18. Isolation and Characterization of Borrelia hermsii Associated with Two Foci of Tick-Borne Relapsing Fever in California

    OpenAIRE

    Fritz, Curtis L; Bronson, Lawrence R.; Smith, Charles R.; Schriefer, Martin E.; Tucker, James R.; Schwan, Tom G.

    2004-01-01

    Relapsing fever, caused by the spirochete Borrelia hermsii and transmitted by the soft tick Ornithodoros hermsi, is endemic in many rural mountainous areas of California. Between 1996 and 1998, 12 cases of relapsing fever associated with two exposure sites in northern California were investigated. Follow-up at exposure sites included collection of soft ticks and serum specimens from sylvatic rodents. Attempts to cultivate spirochetes were made through inoculation of patient blood into mice an...

  19. 5.4.Spirochetosis

    Institute of Scientific and Technical Information of China (English)

    1992-01-01

    920043 Borrelia burgdorferi may be thecausal agent of sarcoidosis.HUA Bing (华冰),et al.Naval Gener Hosp,PLA, Beijing,100037.Chin J Intern Med 1991;30(10):631-633.Serum antibody to Borrelia burgdorferi wasmeasured in 33 patients with sarcoidosis whowere confirmed clinically and pathologically.The

  20. NCBI nr-aa BLAST: CBRC-MDOM-08-0124 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0124 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 5e-26 51% ...

  1. NCBI nr-aa BLAST: CBRC-PHAM-01-1006 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PHAM-01-1006 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 3e-16 42% ...

  2. NCBI nr-aa BLAST: CBRC-MEUG-01-2489 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MEUG-01-2489 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 2e-17 45% ...

  3. NCBI nr-aa BLAST: CBRC-MDOM-01-0368 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0368 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 1e-38 48% ...

  4. NCBI nr-aa BLAST: CBRC-MDOM-08-0235 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-08-0235 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 4e-26 56% ...

  5. NCBI nr-aa BLAST: CBRC-MMUR-01-1360 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MMUR-01-1360 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 1e-19 49% ...

  6. NCBI nr-aa BLAST: CBRC-MDOM-02-0003 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0003 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 2e-19 51% ...

  7. NCBI nr-aa BLAST: CBRC-MDOM-05-0044 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-05-0044 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 8e-14 39% ...

  8. NCBI nr-aa BLAST: CBRC-MDOM-06-0052 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-06-0052 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 5e-23 44% ...

  9. NCBI nr-aa BLAST: CBRC-MDOM-01-0239 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0239 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 2e-35 49% ...

  10. NCBI nr-aa BLAST: CBRC-MDOM-02-0496 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0496 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 4e-26 55% ...

  11. NCBI nr-aa BLAST: CBRC-MDOM-02-0404 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0404 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 8e-24 51% ...

  12. NCBI nr-aa BLAST: CBRC-STRI-01-0275 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-STRI-01-0275 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 3e-04 29% ...

  13. NCBI nr-aa BLAST: CBRC-MDOM-06-0156 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-06-0156 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 2e-37 53% ...

  14. NCBI nr-aa BLAST: CBRC-MDOM-01-0482 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0482 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 2e-31 48% ...

  15. NCBI nr-aa BLAST: CBRC-MDOM-01-0489 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0489 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 7e-29 49% ...

  16. NCBI nr-aa BLAST: CBRC-PHAM-01-1794 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-PHAM-01-1794 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 1e-17 46% ...

  17. NCBI nr-aa BLAST: CBRC-MDOM-03-0296 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-03-0296 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 3e-16 43% ...

  18. NCBI nr-aa BLAST: CBRC-MDOM-01-0081 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0081 ref|YP_002641585.1| hypothetical protein BBU72A_I0017 [Borrelia burg...dorferi 72a] gb|ACM10420.1| putative membrane protein [Borrelia burgdorferi 72a] YP_002641585.1 2e-33 52% ...

  19. NCBI nr-aa BLAST: CBRC-MDOM-02-0399 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0399 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 2e-24 50% ...

  20. NCBI nr-aa BLAST: CBRC-MDOM-01-0482 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-01-0482 ref|YP_002724155.1| hypothetical protein BBU94A_I15 [Borrelia burg...dorferi 94a] gb|ACN92071.1| putative membrane protein [Borrelia burgdorferi 94a] YP_002724155.1 3e-29 49% ...