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Sample records for burgdorferi sensu lato

  1. 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

  2. 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.'

  3. 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.

  4. The ecology of Lyme borreliosis risk : interactions between lxodes ricinus, rodents and Borrelia burgdorferi sensu lato

    NARCIS (Netherlands)

    Duijvendijk, van Gilian

    2016-01-01

    The sheep tick (Ixodes ricinus) is widespread throughout Europe and can transmit Borrelia burgdorferi sensu lato (s.l.), which can cause Lyme borreliosis and B. miyamotoi, the agent of Borrelia miyamotoi disease in humans. Borrelia afzelii is the most common genospeci

  5. Geographical and genospecies distribution of Borrelia burgdorferi sensu lato DNA detected in humans in the USA.

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    Clark, Kerry L; Leydet, Brian F; Threlkeld, Clifford

    2014-05-01

    The present study investigated the cause of illness in human patients primarily in the southern USA with suspected Lyme disease based on erythema migrans-like skin lesions and/or symptoms consistent with early localized or late disseminated Lyme borreliosis. The study also included some patients from other states throughout the USA. Several PCR assays specific for either members of the genus Borrelia or only for Lyme group Borrelia spp. (Borrelia burgdorferi sensu lato), and DNA sequence analysis, were used to identify Borrelia spp. DNA in blood and skin biopsy samples from human patients. B. burgdorferi sensu lato DNA was found in both blood and skin biopsy samples from patients residing in the southern states and elsewhere in the USA, but no evidence of DNA from other Borrelia spp. was detected. Based on phylogenetic analysis of partial flagellin (flaB) gene sequences, strains that clustered separately with B. burgdorferi sensu stricto, Borrelia americana or Borrelia andersonii were associated with Lyme disease-like signs and symptoms in patients from the southern states, as well as from some other areas of the country. Strains most similar to B. burgdorferi sensu stricto and B. americana were found most commonly and appeared to be widely distributed among patients residing throughout the USA. The study findings suggest that human cases of Lyme disease in the southern USA may be more common than previously recognized and may also be caused by more than one species of B. burgdorferi sensu lato. This study provides further evidence that B. burgdorferi sensu stricto is not the only species associated with signs and/or symptoms consistent with Lyme borreliosis in the USA.

  6. 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

  7. 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

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    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.

  8. Detection of tick blood parasites in Egypt using PCR assay II- Borrelia burgdorferi sensu lato.

    Science.gov (United States)

    Adham, Fatma K; El-Samie-Abd, Emtithal M; Gabre, Refaat M; El Hussein, Hala

    2010-12-01

    The prevalence of Borrelia burgdorferi sensu lato (s.l.), the etiologic agent of Lyme borrelosis (LB), was determined for the first time in Egypt by using polymerase chain reaction (PCR). Questing 5243 hard and soft ticks were collected from animal farms throughout Giza Governorate. DNA from 500 individual tick species was extracted and PCR was performed. Primers verified from the sequence of German strain Pko of Borrelia afzelii were used. Fragments of 642 bp were generated and sequenced. The prevalence of B. burgdorferi sensu lato (s.l.) was 28% of examined soft and hard ticks. High infection rate (66%) of B. burgdorferi s.l. was observed in both nymph and adult soft ticks Ornithodoros savignyi. Beside, the role of hard ticks as potential vectors of Lyme disease in Egypt, where the infection rate was between 0.0-50.0%. Sequence analysis of PCR product of Borrelia burgdorferi sensu lato shares high degree of similarity in sequence compared to similar species in GenBank.

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

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    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

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

    Science.gov (United States)

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

    1998-01-01

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

  11. 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...

  12. Real-time PCR-based identification of Borrelia burgdorferi sensu lato species in ticks collected from humans in Romania.

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    Briciu, Violeta T; Meyer, Fabian; Sebah, Daniela; Tăţulescu, Doina F; Coroiu, Georgiana; Lupşe, Mihaela; Carstina, Dumitru; Mihalca, Andrei D; Hizo-Teufel, Cecilia; Klier, Christiane; Huber, Ingrid; Fingerle, Volker

    2014-09-01

    The aims of our study were to determine (i) which tick species bite humans in Romania and (ii) the prevalence of Borrelia (B.) burgdorferi genospecies in these ticks. All ticks collected from patients who presented to the Clinic of Infectious Diseases Cluj Napoca in spring/summer 2010 were morphologically identified by an entomologist and tested for B. burgdorferi genospecies prevalence by a real-time PCR assay targeting the hbb gene and melting curve analysis. Out of 532 ticks, 518 were Ixodes ricinus, 10 Dermacentor marginatus, and 3 Haemaphysalis spp. ticks, and one unidentified tick due to destruction. Since evaluation of the hbb PCR revealed that it was not possible to differentiate between B. spielmanii/B. valaisiana and B. garinii/B. bavariensis, sequencing of an 800-bp fragment of the ospA gene was performed in these cases. Out of 389 investigated ticks, 43 were positive by hbb PCR for B. burgdorferi sensu lato. The positive samples were 42 Ixodes ricinus (11.1% B. burgdorferi sensu lato prevalence) and the one unidentified tick. Species identification revealed the presence of mainly B. afzelii, but also of B. garinii, B. burgdorferi sensu stricto, B. valaisiana, and B. lusitaniae. In 4 samples, differentiation between B. spielmanii/B. valaisiana was impossible. Our study shows that the most relevant human pathogenic B. burgdorferi genospecies - predominantly B. afzelii - are present in ticks collected from Romanian patients.

  13. Occurrence of Bartonella henselae and Borrelia burgdorferi sensu lato co-infections in ticks collected from humans in Germany.

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    Mietze, A; Strube, C; Beyerbach, M; Schnieder, T; Goethe, R

    2011-06-01

    Bartonella (B.) henselae is the zoonotic agent of cat scratch disease. B. henselae has been associated with therapy-resistant Lyme disease in humans suggesting that B. henselae and Borrelia burgdorferi sensu lato might be transmitted concurrently by ticks. In the present study we found that 16 (6.9%) of 230 Ixodes ricinus collected from humans harboured DNA of Bartonella spp. Fifteen positive ticks were infected with B. henselae and one tick with B. clarridgeiae. Twenty-five percent of the 16 Bartonella positive ticks were co-infected with Borrelia burgdorferi sensu lato. Our data show that B. henselae is present in Ixodes ricinus and that ticks may serve as source of infection for humans.

  14. 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.

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

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    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.

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

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    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.

  17. First molecular evidence of [i]Borrelia burgdorferi[/i] sensu lato in goats, sheep, cattle and camels in Tunisia.

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    Ben Said, Mourad; Belkahia, Hanène; Alberti, Alberto; Abdi, Khaoula; Zhioua, Manel; Daaloul-Jedidi, Monia; Messadi, Lilia

    2016-09-01

    Borrelia burgdorferi sensu lato (s.l.) are tick-transmitted spirochaetes of veterinary and human importance. Molecular epidemiology data on ruminants are still lacking in most countries of the world. Therefore, the aim of this study was to estimate the rate of B. burgdorferi s.l. infection in ruminants from Tunisia. A total of 1,021 ruminants (303 goats, 260 sheep, 232 cattle and 226 camels) from different bioclimatic areas in Tunisia were investigated for the presence of B. burgdorferi s.l. DNA in blood by real time PCR. Prevalence rates were 30.4% (92/303) in goats, 6.2% (16/260) in sheep, 1.3% (3/232) in cattle, and 1.8% (4/226) in camels. Only tick species belonging to Rhipicephalus and Hyalomma genera were found on the investigated animals. In small ruminants, the prevalence of B. burgdorferi s.l. varied significantly according to localities and farms. Goats located in humid areas were statistically more infected than those located in sub-humid areas. Prevalence rates varied significantly according to age and breed in sheep, and age and tick infestation in goats. This study provides the first insight into the presence of B. burgdorferi s.l. DNA in ruminants in Tunisia, and demonstrates that host species such as goats and sheep may play an important role in natural Lyme disease cycles in this country.

  18. 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.

  19. Borrelia burgdorferi sensu lato spirochetes in wild birds in northwestern California: associations with ecological factors, bird behavior and tick infestation.

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    Newman, Erica A; Eisen, Lars; Eisen, Rebecca J; Fedorova, Natalia; Hasty, Jeomhee M; Vaughn, Charles; Lane, Robert S

    2015-01-01

    Although Borrelia burgdorferi sensu lato (s.l.) are found in a great diversity of vertebrates, most studies in North America have focused on the role of mammals as spirochete reservoir hosts. We investigated the roles of birds as hosts for subadult Ixodes pacificus ticks and potential reservoirs of the Lyme disease spirochete B. burgdorferi sensu stricto (s.s.) in northwestern California. Overall, 623 birds representing 53 species yielded 284 I. pacificus larvae and nymphs. We used generalized linear models and zero-inflated negative binomial models to determine associations of bird behaviors, taxonomic relationships and infestation by I. pacificus with borrelial infection in the birds. Infection status in birds was best explained by taxonomic order, number of infesting nymphs, sampling year, and log-transformed average body weight. Presence and counts of larvae and nymphs could be predicted by ground- or bark-foraging behavior and contact with dense oak woodland. Molecular analysis yielded the first reported detection of Borrelia bissettii in birds. Moreover, our data suggest that the Golden-crowned Sparrow (Zonotrichia atricapilla), a non-resident species, could be an important reservoir for B. burgdorferi s.s. Of 12 individual birds (9 species) that carried B. burgdorferi s.l.-infected larvae, no birds carried the same genospecies of B. burgdorferi s.l. in their blood as were present in the infected larvae removed from them. Possible reasons for this discrepancy are discussed. Our study is the first to explicitly incorporate both taxonomic relationships and behaviors as predictor variables to identify putative avian reservoirs of B. burgdorferi s.l. Our findings underscore the importance of bird behavior to explain local tick infestation and Borrelia infection in these animals, and suggest the potential for bird-mediated geographic spread of vector ticks and spirochetes in the far-western United States.

  20. Borrelia burgdorferi sensu lato spirochetes in wild birds in northwestern California: associations with ecological factors, bird behavior and tick infestation.

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    Erica A Newman

    Full Text Available Although Borrelia burgdorferi sensu lato (s.l. are found in a great diversity of vertebrates, most studies in North America have focused on the role of mammals as spirochete reservoir hosts. We investigated the roles of birds as hosts for subadult Ixodes pacificus ticks and potential reservoirs of the Lyme disease spirochete B. burgdorferi sensu stricto (s.s. in northwestern California. Overall, 623 birds representing 53 species yielded 284 I. pacificus larvae and nymphs. We used generalized linear models and zero-inflated negative binomial models to determine associations of bird behaviors, taxonomic relationships and infestation by I. pacificus with borrelial infection in the birds. Infection status in birds was best explained by taxonomic order, number of infesting nymphs, sampling year, and log-transformed average body weight. Presence and counts of larvae and nymphs could be predicted by ground- or bark-foraging behavior and contact with dense oak woodland. Molecular analysis yielded the first reported detection of Borrelia bissettii in birds. Moreover, our data suggest that the Golden-crowned Sparrow (Zonotrichia atricapilla, a non-resident species, could be an important reservoir for B. burgdorferi s.s. Of 12 individual birds (9 species that carried B. burgdorferi s.l.-infected larvae, no birds carried the same genospecies of B. burgdorferi s.l. in their blood as were present in the infected larvae removed from them. Possible reasons for this discrepancy are discussed. Our study is the first to explicitly incorporate both taxonomic relationships and behaviors as predictor variables to identify putative avian reservoirs of B. burgdorferi s.l. Our findings underscore the importance of bird behavior to explain local tick infestation and Borrelia infection in these animals, and suggest the potential for bird-mediated geographic spread of vector ticks and spirochetes in the far-western United States.

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

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    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.

  2. 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.

  3. The heterogeneity, distribution and environmental associations of Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, in Scotland

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    Marianne eJames

    2014-08-01

    Full Text Available Lyme borreliosis is an emerging infectious humandisease caused by the Borrelia burgdorferi sensu lato complex of bacteria with reported cases increasing in many areas of Europe and North America. To understand the drivers of disease risk and the distribution of symptoms which may improve mitigation and diagnostics, here we characterise the genetics, distribution and environmental associations of B. burgdorferi s.l. genospecies across Scotland. In Scotland reported Lyme borreliosis cases have increased almost 10-fold since 2000 but the distribution of B. burgdorferi s.l. is so far unstudied. Using a large survey of over 2200 Ixodes ricinus tick samples collected from birds, mammals and vegetation across 25 sites we identified four genospecies: B. afzelii (48%, B. garinii (36% B. valaisiana (8% and B. burgdorferi sensu stricto (7%, and one mixed genospecies infection. Surprisingly, 90% of the sequence types were novel and, importantly, up to 14% of samples were mixed intra-genospecies co-infections, suggesting tick co-feeding, feeding on multiple hosts or multiple infections in hosts. B. garinii (hosted by birds was considerably more genetically diverse than B. afzelii (hosted by small mammals, as predicted since there are more species of birds than small mammals and birds can import strains from mainland Europe. Higher proportions of samples contained B. garinii and B. valaisiana in the west, while B. afzelii and B. garinii were significantly more associated with mixed/deciduous than with coniferous woodlands. This may relate to the abundance of transmission hosts in different regions and habitats. These data on the genetic heterogeneity within and between Borrelia genospecies are a first step to understanding pathogen spread and could help explain the distribution of patient symptoms which may aid local diagnosis. Understanding the environmental associations of the pathogens is critical for rational policy making for disease risk mitigation and

  4. 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 <1400 m asl; I. ricinus was most abundant in Turkey oak (Quercus cerris) wood, whereas H. punctata was mostly collected in hop hornbeam (Ostrya carpinifolia) wood and on exposed rocks. Ixodes ricinus was also found up to 1670 m asl, in high stand beech (Fagus sylvatica) wood. The overall prevalence of 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.

  5. 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...... in squirrels infested with ticks. Invasive grey squirrels appear to become infected with locally circulating strains of B. burgdorferi s.l., further studies are required to determine their role in community disease dynamics. Our findings highlight that the role of introduced host species in B. burgdorferi s...

  6. 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.

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

    DEFF Research Database (Denmark)

    Millins, Caroline; Magierecka, Agnieszka; Gilbert, Lucy

    2015-01-01

    Invasive vertebrate species can act as hosts for endemic pathogens and may alter pathogen community composition and dynamics. For the zoonotic pathogen Borrelia burgdorferi sensu lato, the agent of Lyme borreliosis, recent work shows invasive rodent species can be of high epidemiological importance...... and may support host specific strains. This study examined the role of grey squirrels (Sciurus carolinensis), (n=679), an invasive species in the United Kingdom (UK), as B. burgdorferi s.l. hosts. We found that grey squirrels were frequently infested with Ixodes ricinus, the main vector of B. burgdorferi...... s.l. in the UK, and 11.9% were infected with B. burgdorferi s.l. All four genospecies which occur in the UK were detected in grey squirrels, and unexpectedly, the bird associated genospecies B. garinii, was most common. The second most frequent infection was with B. afzelli. Genotyping of B. garinii...

  8. 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

  9. 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

  10. Molecular analysis of Ixodes granulatus, a possible vector tick for Borrelia burgdorferi sensu lato in Taiwan.

    Science.gov (United States)

    Chao, Li-Lian; Wu, Wen-Jer; Shih, Chien-Ming

    2009-08-01

    The genetic identity of Ixodes granulatus ticks was determined for the first time in Taiwan. The phylogenetic relationships were analyzed by comparing the sequences of mitochondrial 16S ribosomal DNA gene obtained from 19 strains of ticks representing seven species of Ixodes and two outgroup species (Rhipicephalus sanguineus and Haemaphysalis inermis). Four major clades could be easily distinguished by neighbour-joining analysis and were congruent by maximum-parsimony method. All these I. granulatus ticks of Taiwan were genetically affiliated to a monophyletic group with highly homogeneous sequences (92.2-99.3% similarity), and can be discriminated from other Ixodes species and other genera of ticks with a sequence divergence ranging from 11.7 to 30.8%. Moreover, intraspecific analysis revealed that two distinct lineages are evident between the same species of I. granulatus ticks collected from Taiwan and Malaysia. Our results demonstrate that all these I. granulatus ticks of Taiwan represent a unique lineage distinct from the common vector ticks (I. ricinus complex) for Borrelia burgdorferi spirochetes.

  11. 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.

  12. 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

    A multiplex-bead-based assay for the detection of serum antibodies to Borrelia burgdorferi sensu lato was evaluated. The assay contained 13 different antigens in both the IgG and the IgM assay; thus, a total of 26 measurement results were available from each sample. A total of 49 Danish patients...

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

    Directory of Open Access Journals (Sweden)

    Dzięgiel Beata

    2014-03-01

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

  14. 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.

  15. Serological detection of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis antibodies and Dirofilaria immitis antigen in a countrywide survey in dogs in Poland.

    Science.gov (United States)

    Krämer, Friederike; Schaper, Roland; Schunack, Bettina; Połozowski, Andrzej; Piekarska, Jolanta; Szwedko, Aleksandra; Jodies, Robert; Kowalska, Dagmara; Schüpbach, Dörte; Pantchev, Nikola

    2014-09-01

    Canine vector-borne diseases (CVBDs) have increasingly become a focus of attention in the past few years. Nevertheless, in many parts of Europe information on their occurrence is still scarce. In a large study in Poland 3,094 serum samples taken from dogs throughout all 16 Polish provinces were tested using a commercial kit for the detection of circulating antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato and Ehrlichia canis and of Dirofilaria immitis antigen. A total of 12.31% (381/3,094; 95% confidence interval [CI]: 11.18-13.52%) and 3.75% (116/3,094; 95% CI: 3.11-4.48%) of the dogs were positive for A. phagocytophilum and B. burgdorferi s.l. antibodies, respectively. Furthermore, 0.26% (8/3,094; 95% CI: 0.11-0.51%) were positive for E. canis antibodies and 0.16% (5/3,094; 95% CI: 0.05-0.38%) for D. immitis antigen. The highest percentages of A. phagocytophilum-positive dogs were noted in Lesser Poland, Silesia and Łódź Provinces. For B. burgdorferi s.l., the highest prevalence was recorded in Łódź Province. Co-infections with A. phagocytophilum and B. burgdorferi s.l. were recorded in 1.71% of all examined dogs (53/3,094; 95% CI: 1.29-2.23%). One dog even had a triple infection, testing positive for E. canis too. Both A. phagocytophilum and B. burgdorferi s.l. have previously been reported in Poland and were confirmed in the present study by positive samples from all 16 provinces. Concerning E. canis and D. immitis travel history or importation cannot be excluded as factors which may have determined the occurrence of these pathogens in the relevant animals. Practitioners in Poland should be aware of the above mentioned CVBDs and of prophylactic measures to protect dogs and their owners.

  16. First Detection of Borrelia burgdorferi sensu lato DNA in Serum of the Wild Boar (Sus scrofa) in Northern Portugal by Nested-PCR.

    Science.gov (United States)

    Faria, Ana S; Paiva-Cardoso, Maria das Neves; Nunes, Mónica; Carreira, Teresa; Vale-Gonçalves, Hélia M; Veloso, Octávia; Coelho, Catarina; Cabral, João A; Vieira-Pinto, Madalena; Vieira, Maria L

    2015-03-01

    Lyme borreliosis is the most common tick-borne zoonosis in the northern hemisphere. Several vertebrates are crucial in the epidemiological cycle of Borrelia burgdorferi sensu lato, but the role of wild boar as a reservoir is still unknown. Sera were collected from 90 wild boars shot in the Trás-os-Montes region, Northern Portugal (hunting season 2011/2012). In this study, Borrelia DNA was detected for the first time by nested-PCR in three different sera, suggesting that the wild boar may be a potential reservoir for this spirochete. Sequencing results show 100% similarity with Borrelia afzelii. Further studies are needed to evaluate the public health risks associated with boar hunting.

  17. 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

  18. 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.

  19. 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

  20. 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

  1. 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

  2. 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.

  3. Genetic variability within Borrelia burgdorferi sensu lato genospecies established by PCR-single-strand conformation polymorphism analysis of the rrfA-rrlB intergenic spacer in ixodes ricinus ticks from the Czech Republic.

    Science.gov (United States)

    Derdáková, Markéta; Beati, Lorenza; Pet'ko, Branislav; Stanko, Michal; Fish, Durland

    2003-01-01

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

  4. 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

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    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

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

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    Cardoso Luís

    2012-03-01

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

  6. 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.

  7. Seroprevalence and geographic distribution of Dirofilaria immitis and tick-borne infections (Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, and Ehrlichia canis) in dogs from Romania.

    Science.gov (United States)

    Mircean, Viorica; Dumitrache, Mirabela Oana; Györke, Adriana; Pantchev, Nikola; Jodies, Robert; Mihalca, Andrei Daniel; Cozma, Vasile

    2012-07-01

    Tick-borne diseases are of great concern worldwide. Despite this, in Romania there is only limited information regarding the prevalence of vector-borne pathogens in dogs. In all, 1146 serum samples were tested by SNAP(®) 4Dx(®) (IDEXX Laboratories, Inc., Westbrook, ME) for Anaplasma phagocytophilum, Borrelia burgdorferi, and Ehrlichia canis antibodies, and for Dirofilaria immitis antigen. The correlation between positive cases and their geographic distribution, as well as potential risk factors (age, sex, breed, type of dog, habitat, and prophylactic treatments) were evaluated. Overall, 129 dogs (11.3%) were serologically-positive to one or more of the tested pathogens. The seroprevalence for the four infectious agents were: A. phagocytophilum 5.5% (63/1146), D. immitis 3.3% (38/1146), E. canis 2.1% (24/1146), and B. burgdorferi 0.5% (6/1146). Co-infection with E. canis and A. phagocytophilum was registered in 2 dogs (0.2%). The geographical distribution of the seropositive cases suggests clustered foci in southern regions and in the western part of the country for D. immitis, and in the southeastern region (Constanţa County) for E. canis. A. phagocytophilum and B. burgdorferi showed a homogenous distribution, with a tendency for Lyme-positive samples to concentrate in central Romania. For D. immitis, A. phagocytophilum, and E. canis, administering prophylactic treatments was a risk factor associated with infection. Another associated risk factor was the type of dog (stray dogs were at risk being positive for D. immitis, shelter dogs for E. canis, and hunting dogs for B. burgdorferi). The prevalence of D. immitis was significantly higher in males and in dogs older than 2 years. This survey represents the first data detailing A. phagocytophilum and E. canis seroprevalence in Romanian dogs, and the most comprehensive epidemiological study on vector-borne infections in dogs from this country.

  8. Borrelia mayonii sp. nov., a member of the Borrelia burgdorferi sensu lato complex, detected in patients and ticks in the upper midwestern United States.

    Science.gov (United States)

    Pritt, Bobbi S; Respicio-Kingry, Laurel B; Sloan, Lynne M; Schriefer, Martin E; Replogle, Adam J; Bjork, Jenna; Liu, Gongping; Kingry, Luke C; Mead, Paul S; Neitzel, David F; Schiffman, Elizabeth; Hoang Johnson, Diep K; Davis, Jeffrey P; Paskewitz, Susan M; Boxrud, David; Deedon, Alecia; Lee, Xia; Miller, Tracy K; Feist, Michelle A; Steward, Christopher R; Theel, Elitza S; Patel, Robin; Irish, Cole L; Petersen, Jeannine M

    2016-11-01

    Lyme borreliosis (LB) is a multisystem disease caused by spirochetes in the Borrelia burgdorferisensu lato (Bbsl) genospecies complex. We previously described a novel Bbsl genospecies (type strain MN14-1420T) that causes LB among patients with exposures to ticks in the upper midwestern USA. Patients infected with the novel Bbsl genospecies demonstrated higher levels of spirochetemia and somewhat differing clinical symptoms as compared with those infected with other Bbsl genospecies. The organism was detected from human specimens using PCR, microscopy, serology and culture. The taxonomic status was determined using an eight-housekeeping-gene (uvrA, rplB, recG, pyrG, pepX, clpX, clpA and nifS) multi-locus sequence analysis (MLSA) and comparison of 16S rRNA gene, flaB, rrf-rrl, ospC and oppA2 nucleotide sequences. Using a system threshold of 98.3 % similarity for delineation of Bbsl genospecies by MLSA, we demonstrated that the novel species is a member of the Bbsl genospecies complex, most closely related to B. burgdorferisensu stricto (94.7-94.9 % similarity). This same species was identified in Ixodes scapularis ticks collected in Minnesota and Wisconsin. This novel species, Borrelia mayonii sp. nov, is formally described here. The type strain, MN14-1420, is available through the Deutsche Sammlung von Mikroorganismen und Zelkulturen GmbH (DSM 102811) and the American Type Culture Collection (ATCC BAA-2743).

  9. High-rate evolution of Saccharomyces sensu lato chromosomes

    DEFF Research Database (Denmark)

    Spirek, M.; Yang, J.; Groth, C.

    2003-01-01

    Forty isolates belonging to the Saccharomyces sensu lato complex were analyzed for one nuclear and two mitochondrial sequences, and for their karyotypes. These data are useful for description and definition of yeast species based on the phylogenetic species concept. The deduced phylogenetic relat...

  10. Lantana camara L. (sensu lato: an enigmatic complex

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    Neha Goyal

    2015-04-01

    Full Text Available Lantana camara L., considered among the world’s worst invaders is in identity crisis and contentiously referred as Lantana camara L. (sensu lato. Taxonomic ambiguity in L. camara L. (sensu lato, a species complex is one of the grim caveats behind incompetence of its management efforts. Recognizing the extent of variability within the complex, we aim to highlight the need to circumscribe its composition to bring effective management and control efforts into practice. There is a need for clear terminology to examine weedy, naturalized and/or invasive complex constituents that have been placed under the contentious umbrella of ‘L. camara L. (sensu lato’. The time is ripe for invasion ecologists, cytogeneticists and conservationists to collaboratively focus on disentangling the complex and integrate their knowledge and expertise into management and control programs.

  11. Vector Capability of Xiphinema americanum sensu lato in California.

    Science.gov (United States)

    Griesbach, J A; Maggenti, A R

    1989-10-01

    Seven field populations of Xiphinema americanum sensu lato from California's major agronomic areas were tested for their ability to transmit two nepoviruses, including the prune brownline, peach yellow bud, and grapevine yellow vein strains of tomato ringspot virus and the bud blight strain of tobacco ringspot virus. Two field populations transmitted all isolates, one population transmitted all tomato ringspot virus isolates but failed to transmit bud blight strain of tobacco ringspot virus, and the remaining four populations failed to transmit any virus. Only one population, which transmitted all isolates, bad been associated with field spread of a nepovirus. As two California populations of Xiphinema americanum sensu lato were shown to have the ability to vector two different nepoviruses, a nematode taxonomy based on a parsimony of virus-vector relationship is not practical for these populations. Because two California populations of X. americanum were able to vector tobacco ringspot virus, commonly vectored by X. americanum in the eastern United States, these western populations cannot be differentiated from eastern populations by vector capability tests using tobacco ringspot virus.

  12. Antimicrobial activity of Stomoxys calcitransagainst Beauveria bassiana sensu lato isolates

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    Ana Paula Rodrigues de Moraes

    Full Text Available This study had the aims of evaluating the antimicrobial characteristics of Stomoxys calcitrans (Diptera: Muscidae larvae against the fungal isolates CG138, CG228 and ESALQ986 of Beauveria bassiana sensu lato (Balsamo-Crivelli Vuillemin, 1912 (Hypocreales: Cordycipitaceae. S. calcitrans eggs, larvae and pupae were exposed to these same isolates. Statistical analysis showed that the immature stages of S. calcitrans were not susceptible to the fungal isolates used, regardless of the exposure method. Diffusion test on solid culture medium reveled that macerated S. calcitrans larvae exposed to isolate CG138 reduced CG138 fungal development. The analysis of the chromatographic profiles indicated that the macerate or mucus of larvae of the control group and the groups exposed to the isolate CG138 presented different profiles. Reduced development of the isolate CG138 on the larvae cuticle was observed by means of scanning electron microscopy.

  13. Demonstration of Borrelia burgdorferi sensu stricto infection in ticks from the northeast of Mexico.

    Science.gov (United States)

    Gordillo-Pérez, G; Vargas, M; Solórzano-Santos, F; Rivera, A; Polaco, O J; Alvarado, L; Muñóz, O; Torres, J

    2009-05-01

    Borrelia burgdorferisensu lato infection has been confirmed in clinical cases in the northeast of Mexico; however, the bacterium has not been identified as infecting the tick vector Ixodes, Amblyomma and Dermacentor ticks were collected from mammals and plants in northeastern Mexico and examined for Borrelia. Eighteen of 214 ticks were PCR-positive for the fla and 16S rRNA genes and 15 for the ospA gene. Southern blotting with a fla probe and sequencing of ospA genes confirmed infection with B. burgdorferi sensu stricto. These findings, together with reports of indigenous cases, fulfil the criteria that allow northeastern Mexico to be considered as a zone endemic for Lyme disease.

  14. CLASSIFICATION OF GENUS Triticum, SENSU LATO AND SENSU STRICTO, BASED ON SPIKE AND GRAIN MORPHOLOGY

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    Hristo P. STOYANOV

    2015-10-01

    Full Text Available The formulation of the present classifications of species of the genus Triticum associates mainly with several plant morphological factors such as fragility of the spikes spindle, grains threshability, grain sphericity, shape and position of glumes, lemmas and paleas and awns, compactness, etc. Special attention is paid to the factor "cultural/wild" form, the ploidy and the genomic constitution of the species, often supported by molecular data which provides considerable comfort in disclosing phylogenetic features in a particular taxonomic unit. Such taxonomic determination is associated with certain disadvantages. It is not sufficiently focused on the spike morphology related to the reproductive apparatus of the plant, and also the causes of phylogenetic differentiation of certain parameters, such as spike branching, multiple spikelets, as well as the ratios of quantitative properties. The existing classifications do not give a precise answer to the taxonomic position of amphidiploids in the genus Triticum, and also for those obtained from hybrid combinations with genera Aegilops, Secale, Haynaldia, Hordeum, Elymus, Leymus, Elytrigia, Agropyron, as transitional and similar forms. Based on studies of spike and grain morphology of a large number of representatives of the genus Triticum and other interspecific and intergeneric amphidiploid forms, a classification of the genus sensu lato and sensu stricto is composed. Sensu stricto, genus Triticum covers all existing wild and cultivated known wheat forms, together with interspecific artificial synthetic forms. Sensu lato, the genus includes intergeneric hybrids, for which a specific generic epithet was coined - ×Triticum, and also a specific epithet, consistent with the originator of the amphidiploid. Special attention was paid to species and amphidiploids with the genus Aegilops. Classification sensu strictissimo was also formulated where the genus Triticum brings together only diploid species

  15. A proposal for the classification of biological weapons sensu lato.

    Science.gov (United States)

    Rozsa, Lajos

    2014-12-01

    Due to historical and legislation reasons, the category of bioweapons is rather poorly defined. Authors often disagree on involving or excluding agents like hormones, psychochemicals, certain plants and animals (such as weeds or pests) or synthetic organisms. Applying a wide definition apparently threatens by eroding the regime of international legislation, while narrow definitions abandon several important issues. Therefore, I propose a category of 'biological weapons sensu lato' (BWsl) that is defined here as any tool of human aggression whose acting principle is based on disciplines of biology including particularly microbiology, epidemiology, medical biology, physiology, psychology, pharmacology and ecology, but excluding those based on inorganic agents. Synthetically produced equivalents (not necessarily exact copies) and mock weapons are also included. This definition does not involve any claim to subject all these weapons to international legislation but serves a purely scholarly purpose. BWsl may be properly categorized on the base of the magnitude of the human population potentially targeted (4 levels: individuals, towns, countries, global) and the biological nature of the weapons' intended effects (4 levels: agricultural-ecological agents, and non-pathogenic, pathogenic, or lethal agents against humans).

  16. 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

  17. Taxonomy and molecular epidemiology of Echinococcus granulosus sensu lato.

    Science.gov (United States)

    Romig, T; Ebi, D; Wassermann, M

    2015-10-30

    Echinococcus granulosus, formerly regarded as a single species with a high genotypic and phenotypic diversity, is now recognised as an assemblage of cryptic species, which differ considerably in morphology, development, host specificity (including infectivity/pathogenicity for humans) and other aspects. This diversity is reflected in the mitochondrial and nuclear genomes and has led to the construction of phylogenetic trees and hypotheses on the origin and geographic dispersal of various taxa. Based on phenotypic characters and gene sequences, E. granulosus (sensu lato) has by now been subdivided into E. granulosus sensu stricto (including the formerly identified genotypic variants G1-3), Echinococcus felidis (the former 'lion strain'), Echinococcus equinus (the 'horse strain', genotype G4), Echinococcus ortleppi (the 'cattle strain', genotype G5) and Echinococcus canadensis. The latter species, as recognised here, shows the highest diversity and is composed of the 'camel strain', genotype G6, the 'pig strain', genotype G7, and two 'cervid strains', genotypes G8 and G10. There is debate whether the closely related G6 and G7 should be placed in a separate species, but more morphological and biological data are needed to support or reject this view. In this classification, the application of rules for zoological nomenclature led to the resurrection of old species names, which had before been synonymised with E. granulosus. This nomenclatural subdivision of the agents of cystic echinococcosis (CE) may appear inconvenient for practical applications, especially because molecular tools are needed for identification of the cyst stage, and because retrospective data on 'E. granulosus' are now difficult to interpret without examination of voucher specimens. However, the increased awareness for the diversity of CE agents - now emphasised by species names rather than genotype numbers - has led to a large number of recent studies on this issue and a rapid increase of knowledge

  18. Sinopse taxonômica de Boraginaceae sensu lato A. Juss. no Estado de Alagoas, Brasil Taxonomic synopsis of Boraginaceae sensu lato A. Juss. in Alagoas State, Brazil

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    José Iranildo Miranda de Melo

    2008-09-01

    Full Text Available Este trabalho consiste no tratamento sinóptico da família Boraginaceae sensu lato no Estado de Alagoas, Nordeste do Brasil. Foram encontrados três gêneros e 23 espécies: Cordia, com 13 espécies e Heliotropium e Tournefortia, representados por cinco espécies cada. São fornecidas chaves para o reconhecimento de gêneros e espécies, bem como ilustrações, dados de distribuição e hábitat.This work consists of a synoptic treatment for the family Boraginaceae sensu lato in Alagoas state, located in Northeast Brazil. Three genera and 23 species were found: Cordia with 13 species, Heliotropium and Tournefortia with five species each. Keys for determining genera and species are given, as well as illustrations, distribution data and habitat.

  19. Multilocus sequence typing of Pseudomonas syringae sensu lato confirms previously described genomospecies and permits rapid identification.

    Science.gov (United States)

    Since 2002, severe leaf spotting on parsley (Petroselinum crispum L.) has occurred in Monterey County, California. One of two different pathovars of Pseudomonas syringae sensu lato were isolated from diseased leaves from seven distinct outbreaks and twice from the same outbreak (2002 and 2009). Frag...

  20. Selection of Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato isolates as microbial control agents against the boll weevil (Anthonomus grandis) in Argentina.

    Science.gov (United States)

    Nussenbaum, A L; Lecuona, R E

    2012-05-01

    The boll weevil (Anthonomus grandis) is the main pest of cotton in the Americas. The aim of this work was to evaluate isolates of the entomopathogenic fungi Beauveria bassiana sensu lato and Metarhizium anisopliae sensu lato virulent against A. grandis. Screening was performed to evaluate the pathogenicity of 28 isolates of M. anisopliae s.l. and 66 isolates of B. bassiana s.l. against boll weevil adults. To select the isolates, LC(50) values of the most virulent isolates were calculated, and compatibility between the fungi and insecticides was studied. In addition, the effects of these isolates on the feeding behavior of the adults were evaluated. Isolates Ma 50 and Ma 20 were the most virulent against A. grandis and their LC(50) values were 1.13×10(7) and 1.20×10(7) conidia/ml, respectively. In addition, these isolates were compatible with pyrethroid insecticides, but none with endosulfan. On the other hand, infected females reduced the damage caused by feeding on the cotton squares and their weight gain. This shows that entomopathogenic fungi cause mortality in the insects, but also these fungi could influence the feeding behavior of the females. In summary, these results indicate the possibility of the use of M. anisopliae s.l. as a microbiological control agent against boll weevils. Also, this species could be included in an Integrated Pest Management program.

  1. Characterization of virulence properties in the C. parapsilosis sensu lato species.

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    Tibor Németh

    Full Text Available The C. parapsilosis sensu lato group involves three closely related species, C. parapsilosis sensu stricto, C. orthopsilosis and C. metapsilosis. Although their overall clinical importance is dramatically increasing, there are few studies regarding the virulence properties of the species of the psilosis complex. In this study, we tested 63 C. parapsilosis sensu stricto, 12 C. metapsilosis and 18 C. orthopsilosis isolates for the ability to produce extracellular proteases, secrete lipases and form pseudohyphae. Significant differences were noted between species, with the C. metapsilosis strains failing to secrete lipase or to produce pseudohyphae. Nine different clinical isolates each of C. parapsilosis sensu stricto, C. orthopsilosis and C. metapsilosis were co-cultured with immortalized murine or primary human macrophages. C. parapsilosis sensu stricto isolates showed a significantly higher resistance to killing by primary human macrophages compared to C. orthopsilosis and C. metapsilosis isolates. In contrast, the killing of isolates by J774.2 mouse macrophages did not differ significantly between species. However, C. parapsilosis sensu stricto isolates induced the most damage to murine and human macrophages, and C. metapsilosis strains were the least toxic. Furthermore, strains that produced lipase or pseudohyphae were most resistant to macrophage-mediated killing and produced the most cellular damage. Finally, we used 9 isolates of each of the C. parapsilosis sensus lato species to examine their impact on the survival of Galleriamellonella larvae. The mortality rate of G. mellonella larvae infected with C. metapsilosis isolates was significantly lower than those infected with C. parapsilosis sensu stricto or C. orthopsilosis strains. Taken together, our findings demonstrate that C. metapsilosis is indeed the least virulent member of the psilosis group, and also highlight the importance of pseudohyphae and secreted lipases during fungal

  2. Characterization of virulence properties in the C. parapsilosis sensu lato species.

    Science.gov (United States)

    Németh, Tibor; Tóth, Adél; Szenzenstein, Judit; Horváth, Péter; Nosanchuk, Joshua D; Grózer, Zsuzsanna; Tóth, Renáta; Papp, Csaba; Hamari, Zsuzsanna; Vágvölgyi, Csaba; Gácser, Attila

    2013-01-01

    The C. parapsilosis sensu lato group involves three closely related species, C. parapsilosis sensu stricto, C. orthopsilosis and C. metapsilosis. Although their overall clinical importance is dramatically increasing, there are few studies regarding the virulence properties of the species of the psilosis complex. In this study, we tested 63 C. parapsilosis sensu stricto, 12 C. metapsilosis and 18 C. orthopsilosis isolates for the ability to produce extracellular proteases, secrete lipases and form pseudohyphae. Significant differences were noted between species, with the C. metapsilosis strains failing to secrete lipase or to produce pseudohyphae. Nine different clinical isolates each of C. parapsilosis sensu stricto, C. orthopsilosis and C. metapsilosis were co-cultured with immortalized murine or primary human macrophages. C. parapsilosis sensu stricto isolates showed a significantly higher resistance to killing by primary human macrophages compared to C. orthopsilosis and C. metapsilosis isolates. In contrast, the killing of isolates by J774.2 mouse macrophages did not differ significantly between species. However, C. parapsilosis sensu stricto isolates induced the most damage to murine and human macrophages, and C. metapsilosis strains were the least toxic. Furthermore, strains that produced lipase or pseudohyphae were most resistant to macrophage-mediated killing and produced the most cellular damage. Finally, we used 9 isolates of each of the C. parapsilosis sensus lato species to examine their impact on the survival of Galleriamellonella larvae. The mortality rate of G. mellonella larvae infected with C. metapsilosis isolates was significantly lower than those infected with C. parapsilosis sensu stricto or C. orthopsilosis strains. Taken together, our findings demonstrate that C. metapsilosis is indeed the least virulent member of the psilosis group, and also highlight the importance of pseudohyphae and secreted lipases during fungal-host interactions.

  3. Incidence, Antibiotic Susceptibility, and Toxin Profiles of Bacillus cereus sensu lato Isolated from Korean Fermented Soybean Products.

    Science.gov (United States)

    Yim, Jin-Hyeok; Kim, Kwang-Yeop; Chon, Jung-Whan; Kim, Dong-Hyeon; Kim, Hong-Seok; Choi, Da-Som; Choi, In-Soo; Seo, Kun-Ho

    2015-06-01

    Korean fermented soybean products, such as doenjang, kochujang, ssamjang, and cho-kochujang, can harbor foodborne pathogens such as Bacillus cereus sensu lato (B. cereus sensu lato). The aim of this study was to characterize the toxin gene profiles, biochemical characteristics, and antibiotic resistance patterns of B. cereus sensu lato strains isolated from Korean fermented soybean products. Eighty-eight samples of Korean fermented soybean products purchased from retails in Seoul were tested. Thirteen of 26 doenjang samples, 13 of 23 kochujang samples, 16 of 30 ssamjang samples, and 5 of 9 cho-kochujang samples were positive for B. cereus sensu lato strains. The contamination level of all positive samples did not exceed 4 log CFU/g of food (maximum levels of Korea Food Code). Eighty-seven B. cereus sensu lato strains were isolated from 47 positive samples, and all isolates carried at least one enterotoxin gene. The detection rates of hblCDA, nheABC, cytK, and entFM enterotoxin genes among all isolates were 34.5%, 98.9%, 57.5%, and 100%, respectively. Fifteen strains (17.2%) harbored the emetic toxin gene. Most strains tested positive for salicin fermentation (62.1%), starch hydrolysis (66.7%), hemolysis (98.9%), motility test (100%), and lecithinase production (96.6%). The B. cereus sensu lato strains were highly resistant to β-lactam antibiotics such as ampicillin, penicillin, cefepime, imipenem, and oxacillin. Although B. cereus sensu lato levels in Korean fermented soybean products did not exceed the maximum levels permitted in South Korea (<10(4) CFU/g), these results indicate that the bacterial isolates have the potential to cause diarrheal or emetic gastrointestinal diseases.

  4. VERBENACEAE SENSU LATO EM UM TRECHO DA ESEC RASO DA CATARINA, BAHIA, BRASIL

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    JOSÉ IRANILDO MIRANDA DE MELO

    2010-01-01

    Full Text Available This work carried out a floristic-taxonomic survey of the family Verbenaceae sensu lato in a stretch at the Ecological Station Raso da Catarina, Bahia State, Brazil. Four genera and six species were recorded: Aegiphila, with one species (A. sellowiana Cham.; Lantana, with one species (L. fucata Lindl.; Lippia, with three species (L. gracilis Schauer, Lippia cf. schomburgkiana Schauer and L. thymoides Mart. & Schauer and Stachytarpheta, with one species (S. caatingensis S. Atkins. A key for recognition of the species, descriptions and illustrations, beyond data about flowering and fruiting, geographical distribution and habitat are provided.

  5. Differential sensitivity of the species of Candida parapsilosis sensu lato complex against statins.

    Science.gov (United States)

    Szenzenstein, Judit; Gácser, Attila; Grózer, Zsuzsanna; Farkas, Zoltán; Nagy, Katalin; Vágvölgyi, Csaba; Márki-Zay, János; Pfeiffer, Ilona

    2013-10-01

    Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis are human fungal pathogens with clinical importance. The recently reclassified three closely related species have significant variation in virulence, clinical prevalence and susceptibility characteristics to different antifungal compounds. The aim of this study was to investigate the in vitro activity of atorvastatin and fluvastatin against C. metapsilosis, C. orthopsilosis and C. parapsilosis. Susceptibility tests showed that C. parapsilosis was the most sensitive while C. orthopsilosis was the least susceptible species to both drugs. On the basis of the differential sensitivity, we developed a simple, reliable and highly cost-effective plate assay to distinguish these closely related species. Applying this method, 54 isolates belonging to the C. parapsilosis sensu lato complex deposited in Szeged Microbial Collection could be sorted into the three species with 100 % probability.

  6. Ulex parviflorus sensu lato (Genisteae, Leguminosae en la zona centro de Portugal

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    Cubas, Paloma

    1997-06-01

    Full Text Available A group of gorses growing in Central Portugal has been traditionally identified as Ulex parviflorus Pourret sensu lato. The study of these plants reveáls important differences in morphological characters such as size, ramification, colour and indumentum of shoots, the type, disposition and indumentum of primary and secondary spines, and the dimensions, shape and indumentum of the calyx. Based on these morphological characteristics, as well as chromosome number, ecological requirements and geographic distribution, the plants are now ascribed to the following taxa: U. airensis sp. nov., U. australis subsp. welwitschianus stat. & comb. nov., U. eriocladus C. Vicioso and U. jussiaei WebbEn la zona centro de Portugal crece un conjunto de tojos que han sido tradicionalmente identificados como Ulex parviflorus Pourret sensu lato. Sin embargo, estas plantas presentan diferencias morfológicas en el porte, ramificación, color e indumento de ejes y ramas, en el tipo, disposición e indumento de las espinas primarias y secundarias y en las dimensiones, forma e indumento del cáliz. Estos caracteres morfológicos, el número cromosómico, los requerimientos ecológicos y el área geográfica permiten adscribir estas plantas a los siguientes táxones: U. airensis sp. nov., U. australis subsp. welwitschianus stat. & comb. nov., U. eriocladus C. Vicioso y U. jussiaei Webb.

  7. VARIABLES THAT INFLUENCE STUDENTS’ CHOICE OF DISTANCE EDUCATION LATO SENSU GRADUATE BUSINESS PROGRAMS

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    Eduardo Mendes Nascimento

    2014-03-01

    Full Text Available Based on Scriven’s User-Focused Evaluation Theory, the general objective in this study was to identify and analyze the degree of importance Brazilian students attribute to the variables that influence them when choosing distance education lato sensu graduate business programs. The research is classified as descriptive and an electronic questionnaire was used to survey the data, involving 354 students from distance education lato sensu graduate business programs distributed across different Brazilian locations. The questionnaire included 16 variables, which the students were expected to score from 0 to 10. The results indicated that 04 variables obtained a mean score superior to 9, and that flexibility was the main factor the respondents considered in the choice of a distance education program. This evidences that the possibility to structure the program according to their available time is fundamental for the students. Nevertheless, having a trained teaching staff (second most influential variable and a curriculum appropriate to their pedagogical needs (fourth are also essential characteristics. Finally, the respondents indicated the cost as the third most important variable. Some authors even consider it decisive in the students’ choice as distance education programs are frequently cheaper than in-class programs. In addition, it was verified that women score the investigated internal variables higher than men. In addition, the location of the support hub appeared as a determinant variable in the choice of the program.

  8. 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.

  9. Sporothrix schenckii Sensu Lato identification in fragments of skin lesion cultured in NNN medium for differential diagnosis of cutaneous leishmaniasis.

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    Antonio, Liliane de Fátima; Pimentel, Maria Inês Fernandes; Lyra, Marcelo Rosandiski; Madeira, Maria de Fátima; Miranda, Luciana de Freitas Campos; Paes, Rodrigo Almeida; Brito-Santos, Fábio; Carvalho, Maria Helena Galdino Figueredo; Schubach, Armando de Oliveira

    2017-02-01

    Eighty-nine patients with clinical suspicion of leishmaniasis were referred for differential diagnosis. Sporothrix schenckii sensu lato was isolated in Novy-MacNeal-Nicolle + Schneider media in 98% of 64 patients with final diagnosis of sporotrichosis. This medium may be suitable for diagnosis of sporotrichosis in areas where cutaneous leishmaniasis is also endemic.

  10. Genetic diversity of Phytophthora infestans sensu lato in Ecuador provides new insight into the origin of this important plant pathogen

    NARCIS (Netherlands)

    Adler, N.E.; Erselius, L.J.; Chacón, G.M.; Flier, W.G.; Ordonez, M.E.; Kroon, L.P.N.M.; Forbes, G.A.

    2004-01-01

    The metapopulation structure of Phytophthora infestans sensu lato is genetically diverse in the highlands of Ecuador. Previous reports documented the diversity associated with four putative clonal lineages of the pathogen collected from various hosts in the genus Solanum. This paper simultaneously a

  11. Coleopteran and Lepidopteran Hosts of the Entomopathogenic Genus Cordyceps sensu lato

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    Bhushan Shrestha

    2016-01-01

    Full Text Available Entomophthoralean and ascomycetous fungi are the two major groups known to parasitize arthropods in almost every terrestrial habitat of the earth. Within Ascomycota, Cordyceps sensu lato is a large genus with more than 400 spp. described on numerous orders of Arthropoda. Among the hosts of Cordyceps, Coleoptera and Lepidoptera are the two major orders. Out of the estimated 200 Cordyceps spp. recorded on coleopteran and lepidopteran hosts, we have documented 92 spp. based on the available information of their host species. Among coleopteran hosts, Scarabaeidae and Elateridae are the two major families. Similarly, among lepidopterans, Hepialidae is the largest host family. Cordyceps militaris shows the widest host range, extending to 2 orders, 13 families, and 32 spp. We hope such accumulative work will be useful as a quick reference for interested biologists, forest ecologists, biocontrol researchers, and fungal and insect taxonomists to apprehend host range and host specificities of Cordyceps fungi.

  12. A taxonomic revision of the Neoserica (sensu lato calva group (Coleoptera, Scarabaeidae, Sericini

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    Wangang Liu

    2014-10-01

    Full Text Available The species of the Neoserica (sensu lato calva group are revised. Neoserica calva Frey, 1972, comb. n. is redescribed. Thirteen new species are described from China and South Korea: Neoserica ailaoshanica sp. n., N. anonyma sp. n., N. calvoides sp. n., N. gulinqingensis sp. n., N. koelkebecki sp. n., N. liangi sp. n., N. luxiensis sp. n., N. menghaiensis sp. n., N. mengi sp. n., N. taipingensis sp. n., N. zheijangensis sp. n., N. zhibenshanica sp. n., and N. zongyuani sp. n. A key to Sericini genera with multilamellate antenna and species groups of Neoserica of mainland Asia as well as a key to species of the N. calva group are provided. A map of species distribution is given, habitus and male genitalia are illustrated.

  13. Diversity of commensal Bacillus cereus sensu lato isolated from the common sow bug (Porcellio scaber, Isopoda).

    Science.gov (United States)

    Swiecicka, Izabela; Mahillon, Jacques

    2006-04-01

    Although Bacillus cereus sensu lato are important both from an ecological and an economical point of view, little is known about their population structure, ecology, and relationships with other organisms. In the present work, the genotypic similarity of arthropod-borne B. cereus s.l. isolates, and their symbiotic relationship with the host are assessed. Bacilli of this group were recovered from the digestive tracts of sow bugs (Porcellio scaber) collected in three closely located sites. Their genotypic diversity was investigated using pulse-field gel electrophoresis (PFGE) following the whole-genome DNA digestions with NotI and AscI, and PCR amplification of virulence genes. The majority of the sow-bug Bacillus cereus sensu stricto isolates originating from the same but also from different sites displayed identical PFGE patterns, virulence gene content and enterotoxicity, indicating strong genetic and genomic relationships. The sow-bug Bacillus mycoides/Bacillus pseudomycoides strains displayed a higher diversity. The isopod-B. cereus s.l. relationship was also evaluated using antibiotic-resistant derivatives of B. cereus s.s., B. mycoides/B. pseudomycoides and Bacillus thuringiensis reintroduced into sow bugs. Both spores and vegetative cells of B. cereus s.l. were recovered from sow bugs over a 30-day period, strongly suggesting that these bacteria are natural residents of terrestrial isopods.

  14. Candida parapsilosis (sensu lato) isolated from hospitals located in the Southeast of Brazil: Species distribution, antifungal susceptibility and virulence attributes.

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    Ziccardi, Mariangela; Souza, Lucieri O P; Gandra, Rafael M; Galdino, Anna Clara M; Baptista, Andréa R S; Nunes, Ana Paula F; Ribeiro, Mariceli A; Branquinha, Marta H; Santos, André L S

    2015-12-01

    Candida parapsilosis (sensu lato), which represents a fungal complex composed of three genetically related species - Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis, has emerged as an important yeast causing fungemia worldwide. The goal of the present work was to assess the prevalence, antifungal susceptibility and production of virulence traits in 53 clinical isolates previously identified as C. parapsilosis (sensu lato) obtained from hospitals located in the Southeast of Brazil. Species forming this fungal complex are physiologically/morphologically indistinguishable; however, polymerase chain reaction followed by restriction fragment length polymorphism of FKS1 gene has solved the identification inaccuracy, revealing that 43 (81.1%) isolates were identified as C. parapsilosis sensu stricto and 10 (18.9%) as C. orthopsilosis. No C. metapsilosis was found. The geographic distribution of these Candida species was uniform among the studied Brazilian States (São Paulo, Rio de Janeiro and Espírito Santo). All C. orthopsilosis and almost all C. parapsilosis sensu stricto (95.3%) isolates were susceptible to amphotericin B, fluconazole, itraconazole, voriconazole and caspofungin. Nevertheless, one C. parapsilosis sensu stricto isolate was resistant to fluconazole and another one was resistant to caspofungin. C. parapsilosis sensu stricto isolates exhibited higher MIC mean values to amphotericin B, fluconazole and caspofungin than those of C. orthopsilosis, while C. orthopsilosis isolates displayed higher MIC mean to itraconazole compared to C. parapsilosis sensu stricto. Identical MIC mean values to voriconazole were measured for these Candida species. All the isolates of both species were able to form biofilm on polystyrene surface. Impressively, biofilm-growing cells of C. parapsilosis sensu stricto and C. orthopsilosis exhibited a considerable resistance to all antifungal agents tested. Pseudohyphae were observed in 67.4% and 80

  15. Comparative Genomics of Listeria Sensu Lato: Genus-Wide Differences in Evolutionary Dynamics and the Progressive Gain of Complex, Potentially Pathogenicity-Related Traits through Lateral Gene Transfer.

    Science.gov (United States)

    Chiara, Matteo; Caruso, Marta; D'Erchia, Anna Maria; Manzari, Caterina; Fraccalvieri, Rosa; Goffredo, Elisa; Latorre, Laura; Miccolupo, Angela; Padalino, Iolanda; Santagada, Gianfranco; Chiocco, Doriano; Pesole, Graziano; Horner, David S; Parisi, Antonio

    2015-07-15

    Historically, genome-wide and molecular characterization of the genus Listeria has concentrated on the important human pathogen Listeria monocytogenes and a small number of closely related species, together termed Listeria sensu strictu. More recently, a number of genome sequences for more basal, and nonpathogenic, members of the Listeria genus have become available, facilitating a wider perspective on the evolution of pathogenicity and genome level evolutionary dynamics within the entire genus (termed Listeria sensu lato). Here, we have sequenced the genomes of additional Listeria fleischmannii and Listeria newyorkensis isolates and explored the dynamics of genome evolution in Listeria sensu lato. Our analyses suggest that acquisition of genetic material through gene duplication and divergence as well as through lateral gene transfer (mostly from outside Listeria) is widespread throughout the genus. Novel genetic material is apparently subject to rapid turnover. Multiple lines of evidence point to significant differences in evolutionary dynamics between the most basal Listeria subclade and all other congeners, including both sensu strictu and other sensu lato isolates. Strikingly, these differences are likely attributable to stochastic, population-level processes and contribute to observed variation in genome size across the genus. Notably, our analyses indicate that the common ancestor of Listeria sensu lato lacked flagella, which were acquired by lateral gene transfer by a common ancestor of Listeria grayi and Listeria sensu strictu, whereas a recently functionally characterized pathogenicity island, responsible for the capacity to produce cobalamin and utilize ethanolamine/propane-2-diol, was acquired in an ancestor of Listeria sensu strictu.

  16. The optimum cut-off value to differentiate Echinococcus granulosus sensu stricto from other species of E. granulosus sensu lato using larval rostellar hook morphometry.

    Science.gov (United States)

    Soriano, S V; Pierangeli, N B; Pianciola, L A; Mazzeo, M; Lazzarini, L E; Debiaggi, M F; Bergagna, H F J; Basualdo, J A

    2015-01-01

    Cystic echinococcosis caused by Echinococcus granulosus sensu lato is one of the most important helminth zoonoses in the world; it affects both humans and livestock. The disease is endemic in Argentina and highly endemic in the province of Neuquén. Considerable genetic and phenotypic variation has been demonstrated in E. granulosus, and ten different genotypes (G1-G10) have been identified using molecular tools. Echinococcus granulosus sensu lato may be considered a species complex, comprised of E. granulosus sensu stricto (G1-G3), E. equinus (G4), E. ortleppi (G5) and E. canadensis (G6-G10). In endemic areas, the characterization of cystic echinococcosis molecular epidemiology is important in order to apply adequate control strategies. A cut-off value for larval large hook total length to distinguish E. granulosus sensu stricto isolates from those produced by other species of the complex was defined for the first time. Overall, 1780 larval hooks of 36 isolates obtained from sheep (n= 11, G1), goats (n= 10, G6), cattle (n= 5, G6) and pigs (n= 10, G7) were analysed. Validation against molecular genotyping as gold standard was carried out using the receiver operating characteristic (ROC) curve analysis. The optimum cut-off value was defined as 26.5 μm. The proposed method showed high sensitivity (97.8%) and specificity (91.1%). Since in most endemic regions the molecular epidemiology of echinococcosis includes the coexistence of the widely distributed E. granulosus sensu stricto G1 strain and other species of the complex, this technique could be useful as a quick and economical tool for epidemiological and surveillance field studies, when fertile cysts are present.

  17. Echinococcus granulosus sensu lato genotypes infecting humans--review of current knowledge.

    Science.gov (United States)

    Alvarez Rojas, Cristian A; Romig, Thomas; Lightowlers, Marshall W

    2014-01-01

    Genetic variability in the species group Echinococcus granulosus sensu lato is well recognised as affecting intermediate host susceptibility and other biological features of the parasites. Molecular methods have allowed discrimination of different genotypes (G1-10 and the 'lion strain'), some of which are now considered separate species. An accumulation of genotypic analyses undertaken on parasite isolates from human cases of cystic echinococcosis provides the basis upon which an assessment is made here of the relative contribution of the different genotypes to human disease. The allocation of samples to G-numbers becomes increasingly difficult, because much more variability than previously recognised exists in the genotypic clusters G1-3 (=E. granulosus sensu stricto) and G6-10 (Echinococcus canadensis). To accommodate the heterogeneous criteria used for genotyping in the literature, we restrict ourselves to differentiate between E. granulosus sensu stricto (G1-3), Echinococcus equinus (G4), Echinococcus ortleppi (G5) and E. canadensis (G6-7, G8, G10). The genotype G1 is responsible for the great majority of human cystic echinococcosis worldwide (88.44%), has the most cosmopolitan distribution and is often associated with transmission via sheep as intermediate hosts. The closely related genotypes G6 and G7 cause a significant number of human infections (11.07%). The genotype G6 was found to be responsible for 7.34% of infections worldwide. This strain is known from Africa and Asia, where it is transmitted mainly by camels (and goats), and South America, where it appears to be mainly transmitted by goats. The G7 genotype has been responsible for 3.73% of human cases of cystic echinococcosis in eastern European countries, where the parasite is transmitted by pigs. Some of the samples (11) could not be identified with a single specific genotype belonging to E. canadensis (G6/10). Rare cases of human cystic echinococcosis have been identified as having been caused by

  18. Structures of xyloglucans in primary cell walls of gymnosperms, monilophytes (ferns sensu lato) and lycophytes.

    Science.gov (United States)

    Hsieh, Yves S Y; Harris, Philip J

    2012-07-01

    Little is known about the structures of the xyloglucans in the primary cell walls of vascular plants (tracheophytes) other than angiosperms. Xyloglucan structures were examined in 13 species of gymnosperms, 13 species of monilophytes (ferns sensu lato), and two species of lycophytes. Wall preparations were obtained, extracted with 6 M sodium hydroxide, and the extracts treated with a xyloglucan-specific endo-(1→4)-β-glucanase preparation. The oligosaccharides released were analysed by matrix-assisted laser-desorption ionisation time-of-flight mass spectrometry and by high-performance anion-exchange chromatography. The xyloglucan oligosaccharide profiles from the gymnosperm walls were similar to those from the walls of most eudicotyledons and non-commelinid monocotyledons, indicating that the xyloglucans were fucogalactoxyloglucans, containing the fucosylated units XXFG and XLFG. The xyloglucan oligosaccharide profiles for six of the monilophyte species were similar to those of the gymnosperms, indicating they were also fucogalactoxyloglucans. Phylogenetically, these monilophyte species were from both basal and more derived orders. However, the profiles for the other monilophyte species showed various significant differences, including additional oligosaccharides. In three of the species, these additional oligosaccharides contained arabinosyl residues which were most abundant in the profile of Equisetum hyemale. The two species of lycophytes examined, Selaginella kraussiana and Lycopodium cernuum, had quite different xyloglucan oligosaccharide profiles, but neither were fucogalactoxyloglucans. The S. kraussiana profile had abundant oligosaccharides containing arabinosyl residues. The L. cernuum profile indicated the xyloglucan had a very complex structure.

  19. Update of phylogenetic and genetic diversity of Sporothrix schenckii sensu lato.

    Science.gov (United States)

    Rangel-Gamboa, Lucía; Martínez-Hernandez, Fernando; Maravilla, Pablo; Arenas-Guzmán, Roberto; Flisser, Ana

    2016-03-01

    Sporothrix schenckii sensu lato causes subcutaneous mycosis. In this article we analysed its phylogeny and genetic diversity using calmodulin DNA sequences deposited in GenBank database. Population genetics indices were calculated, plus phylogenetic and haplotype network trees were built. Five clades with high values of posterior probability, 47 haplotypes and high diversity in the complex were found. Analysis of partial calmodulin sequences alignment revealed conserved and polymorphic regions that could be used as reference for taxonomic identification. The use of population genetics analysis allowed understanding the phylogenetic proximity of S. schenckii s. str. and S. brasiliensis; scarce genetic flow among them with low migration index and high ancestry coefficient was found. Similarly, S. globosa, S. mexicana and S. pallida sequences showed highly differentiated species with no genetic exchange. The phylogenetic tree suggests that S. mexicana shared a common ancestor with S. pallida; while S. globosa and S. brasiliensis are more related to S. schenckii s. str. and showed less haplotype diversity and restrictions in geographic distribution. In the haplotype network tree S. schenckii s. str. species displayed worldwide distribution without dispersion centres; while S. brasiliensis and S. globosa, exhibited Brazil and Euro-Asia as dispersion centres, respectively. Our data suggest that S. schenckii complex has been submitted to a divergent evolution process, probably due to the pressure of the environment and of the host. In contrast, S. brasiliensis could have been submitted to purifying selection or expansion process.

  20. The order Corallinales sensu lato (Rhodophyta in the Iberian Atlantic: current state of knowledge

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    Juan Lugilde

    2016-09-01

    Full Text Available A review of the order Corallinales sensu lato in the Atlantic Iberian Peninsula is presented with the aim of assessing its current state of knowledge in comparison with adjacent areas (British Isles-Atlantic France, Macaronesia and Iberian Mediterranean. According to the information compiled from more than 250 publications, herbarium data and manuscripts, we concluded that Atlantic Iberian coralline algae have been poorly studied, which resulted in only 49 species reported. By contrast, Macaronesia is the most species-rich region (91, followed by Spanish Mediterranean (67 and the British Isles-Atlantic France (61. In the Atlantic Iberian Peninsula, 17 species occurred commonly (present in more than 50% of the coastline, particularly those corresponding to the genera Amphiroa, Jania, Lithophyllum, Mesophyllum, and Phymatolithon. Instead, the genera Harveylithon, Hydrolithon, Leptophytum, Lithothamnion, Neogoniolithon and Pneophyllum have been occasionally reported. In the Atlantic Iberian Peninsula and adjacent regions, the epilithic growth-form was dominant, followed by the epiphytic, epizoic and the unattached (maerl/rodoliths; besides, sciaphilous taxa were more abundant than photophilous species. The low intertidal and shallow subtidal harbour a high diversity of coralline algae, as well as semi-exposed coasts or areas affected by currents. The present study confirms that studies on the Atlantic Iberian coralline algae are scarce, and that further research on this group is required.

  1. Assessment of the efficacy of orally administered afoxolaner against Rhipicephalus sanguineus sensu lato.

    Science.gov (United States)

    Kunkle, Bruce; Daly, Sean; Dumont, Pascal; Drag, Marlene; Larsen, Diane

    2014-04-02

    Two studies were conducted to confirm that a single oral dose of the novel insecticide/acaricide afoxolaner is efficacious against existing infestations of Rhipicephalus sanguineus sensu lato in dogs and can control re-infestation for up to 35 days. Each study utilized 16 purpose bred adult dogs using a controlled randomized block design. One or two days prior to treatment, all dogs were infested with 50 unfed adult ticks. On Day 0 one group was treated with an oral chewable formulation of afoxolaner at a dose as close as possible to the minimum dose of 2.5mg/kg. Weekly re-infestations with 50 adult unfed ticks were repeated for five weeks. Forty-eight hours after treatment and after each re-infestation, the number of remaining live ticks on each dog was counted. Treatment with afoxolaner resulted in efficacies of 98.8-100% within 48 h on existing tick infestations, while the efficacy against new tick infestations was >95.7% over five weeks.

  2. Retreat and extinction of the Late Pleistocene cave bear ( Ursus spelaeus sensu lato)

    Science.gov (United States)

    Baca, Mateusz; Popović, Danijela; Stefaniak, Krzysztof; Marciszak, Adrian; Urbanowski, Mikołaj; Nadachowski, Adam; Mackiewicz, Paweł

    2016-12-01

    The cave bear ( Ursus spelaeus sensu lato) is a typical representative of Pleistocene megafauna which became extinct at the end of the Last Glacial. Detailed knowledge of cave bear extinction could explain this spectacular ecological transformation. The paper provides a report on the youngest remains of the cave bear dated to 20,930 ± 140 14C years before present (BP). Ancient DNA analyses proved its affiliation to the Ursus ingressus haplotype. Using this record and 205 other dates, we determined, following eight approaches, the extinction time of this mammal at 26,100-24,300 cal. years BP. The time is only slightly earlier, i.e. 27,000-26,100 cal. years BP, when young dates without associated collagen data are excluded. The demise of cave bear falls within the coldest phase of the last glacial period, Greenland Stadial 3. This finding and the significant decrease in the cave bear records with cooling indicate that the drastic climatic changes were responsible for its extinction. Climate deterioration lowered vegetation productivity, on which the cave bear strongly depended as a strict herbivore. The distribution of the last cave bear records in Europe suggests that this animal was vanishing by fragmentation into subpopulations occupying small habitats. One of them was the Kraków-Częstochowa Upland in Poland, where we discovered the latest record of the cave bear and also two other, younger than 25,000 14C years BP. The relatively long survival of this bear in karst regions may result from suitable microclimate and continuous access to water provided by deep aquifers, indicating a refugial role of such regions in the Pleistocene for many species.

  3. Synopsis of Boraginaceae sensu lato in the Caatingas of the São Francisco River, Northeastern Brazil

    Directory of Open Access Journals (Sweden)

    Miranda de Melo, José Iranildo

    2015-06-01

    Full Text Available This work presents a commented synopsis of Boraginaceae sensu lato in the basin of the lower-middle and lower stretches of the São Francisco River in Northeast Brazil (states of Pernambuco and Bahia. Five genera and 21 species were recorded: Euploca, with six species; Cordia, with five; Varronia, with four; and Heliotropium and Myriopus, represented by three species each. A new combination in the genus Myriopus is here proposed. Keys for the separation of genera and species as well as distributional and geographical data are provided.Este trabajo presenta una sinopsis comentada de la familia Boraginaceae sensu lato en la cuenca media y baja del río São Francisco, nordeste del Brasil (estados de Pernambuco y Bahía. Se reconocen cinco géneros y 21 especies: Euploca, con seis especies; Cordia, con cinco; Varronia, con cuatro y Heliotropium y Myriopus, representados por tres especies cada. Se propone una combinación nueva en el género Myriopus. Así mismo, se aportan claves para la separación de los géneros y especies y datos sobre la distribución geográfica y la ecología de las especies.

  4. Brown dog tick, Rhipicephalus sanguineus sensu lato, infestation ofsusceptible dog hosts is reduced by slow release of semiochemicalsfrom a less susceptible host

    Science.gov (United States)

    Domestic dog breeds are hosts for the tick Rhipicephalus sanguineus sensu lato, but infestation levels vary among breeds. Beagles are less susceptible to tick infestations than English cocker spaniels due to enhanced production of 2-hexanone and benzaldehyde that act as tick repellents. We report th...

  5. First molecular evidence of the simultaneous human infection with two species of Echinococcus granulosus sensu lato: Echinococcus granulosus sensu stricto and Echinococcus canadensis.

    Science.gov (United States)

    Oudni-M'rad, Myriam; M'rad, Selim; Ksia, Amine; Lamiri, Rachida; Mekki, Mongi; Nouri, Abdellatif; Mezhoud, Habib; Babba, Hamouda

    2016-03-01

    Cystic echinococcosis is a widespread zoonotic parasitic disease especially in Tunisia which is one of the most endemic countries in the Mediterranean area. The etiological agent, Echinococcus granulosus sensu lato, implies dogs and other canids as definitive hosts and different herbivore species as intermediate hosts. Human contamination occurs during the consumption of parasite eggs passed in the environment through canid feces. Hydatid cysts coming from a child operated for multiple echinococcosis were collected and analyzed in order to genotype and to obtain some epidemiological molecular information. Three targets, ribosomal DNA ITS1 fragment, NADH dehydrogenase subunit 1 (nad1), and mitochondrial cytochrome c oxydase subunit 1 (CO1) genes, were amplified and analyzed by RFLP and sequencing approach. This study presents the first worldwide report in human of a simultaneous infection with Echinococcus granulosus sensu stricto (genotype G1) and Echinococcus canadensis (genotype G6) species. This is also the first report of the presence of E. canadensis in the Tunisian population which argues in favor of a greater importance of this species in human infestation in Tunisia than previously believed.

  6. Karyomorphology of six taxa in Chrysanthemum sensu lato (Anthemideae) in Egypt and their genetic relationships by Giemsa C-banding

    Institute of Scientific and Technical Information of China (English)

    Magdy Hussein ABD EL-TWAB; Ahmad Mohammad M. MEKAWY; Mohammad Saad EL-KATATNY

    2012-01-01

    Giemsa C-banding was applied to the chromosome complements of six diploid species belonging to six genera in Chrysanthemum sensu lato (Anthemideae) distributed in Egypt.Four types of C-banding distribution were observed in the taxa as follows:(i) negative C-banding in Anacyclus monanthos (L.) Thell.; (ii) all bands in terminal regions in Achilleafragrantissima (Forssk.) Sch.Bip,which showed 32 bands on 18 chromosomes; (iii)all eight bands at centromeric regions on eight chromosomes in Matricaria recutita L.; and (iv) bands at terminal and centromeric regions in Brocchia cinerea Vis.(12 terminal and six centromeric bands on 12 chromosomes),Cotula barbata DC.(four terminal,six centromeric,and eight short arm bands on 16 chromosomes),and Glebionis coronaria (L.) Cass.ex Spach.(eight terminal on the short arms and four large bands in centromeric regions on 12chromosomes).

  7. The Identification of Intrinsic Chloramphenicol and Tetracycline Resistance Genes in Members of the Bacillus cereus Group (sensu lato)

    Science.gov (United States)

    Glenwright, Helen; Pohl, Susanne; Navarro, Ferran; Miro, Elisenda; Jiménez, Guillermo; Blanch, Anicet R.; Harwood, Colin R.

    2017-01-01

    Bacillus toyonensis strain BCT-7112T (NCIMB 14858T) has been widely used as an additive in animal nutrition for more than 30 years without reports of adverse toxigenic effects. However, this strain is resistant to chloramphenicol and tetracycline and it is generally considered inadvisable to introduce into the food chain resistance determinants capable of being transferred to other bacterial strains, thereby adding to the pool of such determinants in the gastro-enteric systems of livestock species. We therefore characterized the resistance phenotypes of this strain and its close relatives to determine whether they were of recent origin, and therefore likely to be transmissible. To this end we identified the genes responsible for chloramphenicol (catQ) and tetracycline (tetM) resistance and confirmed the presence of homologs in other members of the B. toyonensis taxonomic unit. Unexpectedly, closely related strains encoding these genes did not exhibit chloramphenicol and tetracycline resistance phenotypes. To understand the differences in the behaviors, we cloned and expressed the genes, together with their upstream regulatory regions, into Bacillus subtilis. The data showed that the genes encoded functional proteins, but were expressed inefficiently from their native promoters. B. toyonensis is a taxonomic unit member of the Bacillus cereus group (sensu lato). We therefore extended the analysis to determine the extent to which homologous chloramphenicol and tetracycline resistance genes were present in other species within this group. This analysis revealed that homologous genes were present in nearly all representative species within the B. cereus group (sensu lato). The absence of known transposition elements and the observations that they are found at the same genomic locations, indicates that these chloramphenicol and tetracycline resistance genes are of ancient origin and intrinsic to this taxonomic group, rather than recent acquisitions. In this context we

  8. The Identification of Intrinsic Chloramphenicol and Tetracycline Resistance Genes in Members of the Bacillus cereus Group (sensu lato).

    Science.gov (United States)

    Glenwright, Helen; Pohl, Susanne; Navarro, Ferran; Miro, Elisenda; Jiménez, Guillermo; Blanch, Anicet R; Harwood, Colin R

    2016-01-01

    Bacillus toyonensis strain BCT-7112(T) (NCIMB 14858(T)) has been widely used as an additive in animal nutrition for more than 30 years without reports of adverse toxigenic effects. However, this strain is resistant to chloramphenicol and tetracycline and it is generally considered inadvisable to introduce into the food chain resistance determinants capable of being transferred to other bacterial strains, thereby adding to the pool of such determinants in the gastro-enteric systems of livestock species. We therefore characterized the resistance phenotypes of this strain and its close relatives to determine whether they were of recent origin, and therefore likely to be transmissible. To this end we identified the genes responsible for chloramphenicol (catQ) and tetracycline (tetM) resistance and confirmed the presence of homologs in other members of the B. toyonensis taxonomic unit. Unexpectedly, closely related strains encoding these genes did not exhibit chloramphenicol and tetracycline resistance phenotypes. To understand the differences in the behaviors, we cloned and expressed the genes, together with their upstream regulatory regions, into Bacillus subtilis. The data showed that the genes encoded functional proteins, but were expressed inefficiently from their native promoters. B. toyonensis is a taxonomic unit member of the Bacillus cereus group (sensu lato). We therefore extended the analysis to determine the extent to which homologous chloramphenicol and tetracycline resistance genes were present in other species within this group. This analysis revealed that homologous genes were present in nearly all representative species within the B. cereus group (sensu lato). The absence of known transposition elements and the observations that they are found at the same genomic locations, indicates that these chloramphenicol and tetracycline resistance genes are of ancient origin and intrinsic to this taxonomic group, rather than recent acquisitions. In this context

  9. 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

  10. Early cytokine release in response to live Borrelia burgdorferi Sensu Lato Spirochetes is largely complement independent.

    Directory of Open Access Journals (Sweden)

    Kerstin Sandholm

    Full Text Available AIM: Here we investigated the role of complement activation in phagocytosis and the release of cytokines and chemokines in response to two clinical isolates: Borrelia afzelii K78, which is resistant to complement-mediated lysis, and Borrelia garinii LU59, which is complement-sensitive. METHODS: Borrelia spirochetes were incubated in hirudin plasma, or hirudin-anticoagulated whole blood. Complement activation was measured as the generation of C3a and sC5b-9. Binding of the complement components C3, factor H, C4, and C4BP to the bacterial surfaces was analyzed. The importance of complement activation on phagocytosis, and on the release of cytokines and chemokines, was investigated using inhibitors acting at different levels of the complement cascade. RESULTS: 1 Borrelia garinii LU59 induced significantly higher complement activation than did Borrelia afzelii K78. 2 Borrelia afzelii K78 recruited higher amounts of factor H resulting in significantly lower C3 binding. 3 Both Borrelia strains were efficiently phagocytized by granulocytes and monocytes, with substantial inhibition by complement blockade at the levels of C3 and C5. 4 The release of the pro-inflammatory cytokines and chemokines IL-1β, IL-6, TNF, CCL20, and CXCL8, together with the anti-inflammatory IL-10, were increased the most (by>10-fold after exposure to Borrelia. 5 Both strains induced a similar release of cytokines and chemokines, which in contrast to the phagocytosis, was almost totally unaffected by complement blockade. CONCLUSIONS: Our results show that complement activation plays an important role in the process of phagocytosis but not in the subsequent cytokine release in response to live Borrelia spirochetes.

  11. Pteridologische aantekeningen, 3, De eikvaren (Polypodium vulgare L. sensu lato) in Nederland

    NARCIS (Netherlands)

    Meinders-Groeneveld, J.; Segal, S.

    1967-01-01

    Pteridological Notes, 3 The genus Polypodium in the Netherlands Both Polypodium vulgare L. (sensu stricto) and P. interjectum Shivas occur in the Netherlands. Most of the differential characters reported in the literature overlap one another rather broadly, so that it is often necessary to use a ’sy

  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. A comparative laboratory trial evaluating the immediate efficacy of fluralaner, afoxolaner, sarolaner and imidacloprid + permethrin against adult Rhipicephalus sanguineus (sensu lato) ticks attached to dogs

    OpenAIRE

    Burgio, Federica; Meyer, Leon; Armstrong, Rob

    2016-01-01

    Background Acaricides are used to treat and prevent tick infestations, and a common clinical scenario is to administer an acaricide on observing an attached tick. Consequently, immediate acaricidal efficacy (onset of activity and speed of kill) results are clinically valuable. This study evaluated the immediate efficacy of four commercially available acaricides against adult Rhipicephalus sanguineus (sensu lato). Methods Forty dogs were blocked on hair length and tick carrying capacity, then ...

  14. Comparative transcriptional profiling of Bacillus cereus sensu lato strains during growth in CO2-bicarbonate and aerobic atmospheres.

    Directory of Open Access Journals (Sweden)

    Karla D Passalacqua

    Full Text Available BACKGROUND: Bacillus species are spore-forming bacteria that are ubiquitous in the environment and display a range of virulent and avirulent phenotypes. This range is particularly evident in the Bacillus cereus sensu lato group; where closely related strains cause anthrax, food-borne illnesses, and pneumonia, but can also be non-pathogenic. Although much of this phenotypic range can be attributed to the presence or absence of a few key virulence factors, there are other virulence-associated loci that are conserved throughout the B. cereus group, and we hypothesized that these genes may be regulated differently in pathogenic and non-pathogenic strains. METHODOLOGY/PRINCIPAL FINDINGS: Here we report transcriptional profiles of three closely related but phenotypically unique members of the Bacillus cereus group--a pneumonia-causing B. cereus strain (G9241, an attenuated strain of B. anthracis (Sterne 34F(2, and an avirulent B. cereus strain (10987--during exponential growth in two distinct atmospheric environments: 14% CO(2/bicarbonate and ambient air. We show that the disease-causing Bacillus strains undergo more distinctive transcriptional changes between the two environments, and that the expression of plasmid-encoded virulence genes was increased exclusively in the CO(2 environment. We observed a core of conserved metabolic genes that were differentially expressed in all three strains in both conditions. Additionally, the expression profiles of putative virulence genes in G9241 suggest that this strain, unlike Bacillus anthracis, may regulate gene expression with both PlcR and AtxA transcriptional regulators, each acting in a different environment. CONCLUSIONS/SIGNIFICANCE: We have shown that homologous and even identical genes within the genomes of three closely related members of the B. cereus sensu lato group are in some instances regulated very differently, and that these differences can have important implications for virulence. This study

  15. Révision systématique des Hipparion sensu lato (Perissodactyla, Equidae de l'Ancien Monde

    Directory of Open Access Journals (Sweden)

    Zouhri, S.

    2005-04-01

    Full Text Available La systématique des Hipparion sensu lato de l’Ancien Monde se caractérise par un foisonnement anormal des espèces rapportées à ce groupe et par une vive controverse sur le statut supraspécifique de celles-ci d’où une confusion générale de la phylogénie de ce groupe et bien évidemment des interprétations biochronologiques et biogéographiques qui en découlent. L’analyse critique de la systématique de chacune de ces nombreuses espèces d’Hipparion [s. l.] a permis une discussion et une réévaluation de la validité de ces dernières et la suggestion de synonymies et de regroupements de certaines espèces et l’élaboration d’une nouvelle classification supraspécifique de ce groupe. Quatre genres ont été alors redéfinis : Hippotherium, Hipparion, Cremohipparion et Proboscidipparion. Ce dernier taxon est subdivisé en trois sous-genres : Proboscidipparion, Plesiohipparion et Eurygnathohippus.La sistemática de Hipparion sensu lato del Antiguo Mundo se caracteriza por una multiplicación excesiva de especies relacionadas con este grupo y una marcada controversia sobre el status supraespecífico que trae consigo una confusión general de su filogenia, así como de las interpretaciones biocronológicas y biogeográficas que se deducen. El análisis crítico de la sistemática de cada una de las numerosas especies de Hipparion [s. l.] del Antiguo Mundo ha permitido una discusión y una reevaluación de la validez de las mismas, así como una nueva clasificación supraespecífica de este grupo. Cuatro géneros han sido redefinidos: Hippotherium, Hipparion, Cremohipparion y Proboscidipparion. Este último taxon ha sido subdividido en tres subgéneros: Proboscidipparion, Plesiohipparion y Eurygnathohippus. La validez, las sinonimias y los reagrupamientos de las especies relacionadas con cada uno de estos taxones han sido discutidos y analizados en detalle.

  16. Environmental Contamination by Echinococcus granulosus sensu lato Eggs in Relation to Slaughterhouses in Urban and Rural Areas in Tunisia.

    Science.gov (United States)

    Chaâbane-Banaoues, Raja; Oudni-M'rad, Myriam; M'rad, Selim; Mezhoud, Habib; Babba, Hamouda

    2016-02-01

    Hydatidosis has become a real concern for health care institutions and animal rearers in Tunisia. The Tunisian endemicity is aggravated by the growing number of dogs and the difficulty of getting rid of contaminated viscera because of the lack of equipment in most slaughterhouses. Therefore, microscopic and molecular tools were applied to evaluate the role of slaughterhouses in canine infection and Echinococcus granulosus sensu lato (s. l.) egg dissemination. Exposure risk to E. granulosus s. l. eggs in urban and rural areas was explored in order to implant preventive and adapted control strategies. Microscopic examinations detected taeniid eggs in 152 amongst 553 fecal samples. The copro-PCR demonstrated that 138 of 152 taeniid samples analyzed were positive for E. granulosus s. l. DNA. PCR-RFLP demonstrated that all isolated samples belonged to E. granulosus sensu stricto (s. s.). An important environmental contamination index (25.0%) by E. granulosus s. l. eggs was demonstrated. The average contamination index from the regions around slaughterhouses (23.3%; 95% CI: 17.7-28.9%) was in the same range as detected in areas located far from slaughterhouses (26.0%, 95% CI: 21.3-30.8%). Echinococcosis endemic areas were extended in both rural (29.9%, 95% CI: 24.8-34.9%) and urban locations (18.1%, 95% CI: 13.0-22.9%). The pathogen dissemination is related neither to the presence/absence of slaughterhouses nor to the location in urban or rural areas, but is probably influenced by human activities (home slaughtering) and behavior towards the infected viscera.

  17. Changing patterns of prevalence in Opisthorchis viverrini sensu lato infection in children and adolescents in northeast Thailand.

    Science.gov (United States)

    Khuntikeo, Narong; Sithithaworn, Paiboon; Loilom, Watcharin; Namwat, Nisana; Yongvanit, Puangrat; Thinkhamrop, Bandit; Kiatsopit, Nadda; Andrews, Ross H; Petney, Trevor N

    2016-12-01

    Infection with the liver fluke Opisthorchis viverrini sensu lato (s.l.), a group 1 carcinogen, is the most important risk factor for developing cholangiocarcinoma (CCA) in Southeast Asia. Cholangiocarcinoma is a fatal disease with the world's highest incidence being found in northeast Thailand. Liver fluke infection occurs through eating raw or partially cooked cyprinid fish containing metacercariae and, therefore, the control of O. viverrini s.l. infection should lead to a reduction in CCA incidence. In this report, we review and analyze the age-prevalence profile data of O. viverrini to reveal temporal changes in patterns of prevalence pre- and post-control programs in Thailand. The profiles of O. viverrini prevalence have transformed from high prevalence in school children prior to 1983 to low prevalences after 1994. This pattern strongly suggests the influence of the health education program on the likelihood of school children becoming infected. In conjunction with current developments in health and socioeconomic conditions, we predict that the incidence of CCA will be reduced with time as the population cohorts that experienced the education programs reach the age at which CCA is most likely to develop, i.e. >50 years. The lessons learned in Thailand may be applicable to other areas endemic for human liver flukes.

  18. Seasonal transmission of Opisthorchis viverrini sensu lato and a lecithodendriid trematode species in Bithynia siamensis goniomphalos snails in northeast Thailand.

    Science.gov (United States)

    Namsanor, Jutamas; Sithithaworn, Paiboon; Kopolrat, Kulthida; Kiatsopit, Nadda; Pitaksakulrat, Opal; Tesana, Smarn; Andrews, Ross H; Petney, Trevor N

    2015-07-01

    Seasonal changes play roles in the transmission success of fish-borne zoonotic trematodes (FZT). This study examined the seasonal transmission patterns of Opisthorchis viverrini sensu lato (s.l.) and a virgulate cercaria (family Lecithodendriidae) in the snail intermediate host, Bithynia siamensis goniomphalos in northeast Thailand. Snail samples were collected monthly during the rainy, cool, and hot seasons during 2012-2013 to determine the prevalence and intensity of larval trematode infections. The prevalence of O. viverrini s.l. varied significantly with season, being 0.31%, 1.05%, and 0.37% in the rainy, cool, and hot seasons, respectively (P < 0.05). Similarly, the prevalence of virgulate cercariae was 3.11%, 6.80%, and 1.64% in the rainy, cool, and hot seasons, respectively (P < 0.05). The intensity of larval trematode infections also varied between seasons and peaked in the hot season (P < 0.05) in both species. The snails infected with O. viverrini s.l. were significantly smaller (P < 0.05) and those infected with virgulate cercariae were significantly larger (P < 0.05) than uninfected snails. Seasonal variation and the different sizes of B. s. goniomphalos parasitized by O. viverrini s.l. and virgulate trematodes indicate complex host-parasite interactions with important implications for the epidemiology of O. viverrini s.l.

  19. The systematics and population genetics of Opisthorchis viverrini sensu lato: implications in parasite epidemiology and bile duct cancer.

    Science.gov (United States)

    Sithithaworn, Paiboon; Andrews, Ross H; Petney, Trevor N; Saijuntha, Weerachai; Laoprom, Nonglak

    2012-03-01

    Together with host and environmental factors, the systematics and population genetic variation of Opisthorchis viverrini may contribute to recorded local and regional differences in epidemiology and host morbidity in opisthorchiasis and cholangiocarcinoma (CCA). In this review, we address recent findings that O. viverrini comprises a species complex with varying degrees of population genetic variation which are associated with specific river wetland systems within Thailand as well as the Lao PDR. Having an accurate understanding of systematics is a prerequisite for a meaningful assessment of the population structure of each species within the O. viverrini complex in nature, as well as a better understanding of the magnitude of genetic variation that occurs within different species of hosts in its life cycle. Whether specific genotypes are related to habitat type(s) and/or specific intermediate host species are discussed based on current available data. Most importantly, we focus on whether there is a correlation between incidence of CCA and genotype(s) of O. viverrini. This will provide a solid basis for further comprehensive investigations of the role of genetic variation within each species of O. viverrini sensu lato in human epidemiology and genotype related morbidity as well as co-evolution of parasites with primary and secondary intermediate species of host.

  20. Use of FTA(®) card methodology for sampling and molecular characterization of Echinococcus granulosus sensu lato in Africa.

    Science.gov (United States)

    Boué, Franck; El Berbri, Ikhlass; Hormaz, Vanessa; Boucher, Jean-Marc; El Mamy, Ahmed Bezeid; Traore, Abdallah; Fihri, Ouafaa Fassi; Petavy, Anne-Françoise; Dakkak, Allal; Umhang, Gérald

    2017-02-01

    Cystic Echinococcosis is a parasitic disease caused by the cestode Echinococcus granulosus widely distributed in Africa. Monitoring of this parasite requires access to cyst samples on intermediate hosts observed at the slaughterhouse. In order to facilitate sampling in the field and analysis, the French National Reference Laboratory for Echinococcus spp. has developed a tissue derived from DNA sampling with FTA(®) card technology. The DNA samples were taken by applying the FTA(®) paper on the germinal layer after opening the cysts. The sampling technique was validated using frozen cysts (n = 76) stored in the laboratory and from field samples (n = 134) taken at the slaughterhouse by veterinarian technicians during meat inspection in Morocco, Mali and Mauritania. DNA was extracted after several weeks of storage at room temperature. PCR assays were performed using primers for generic cestode (cox1) and amplified fragments were sequenced. All samples taken in the lab and 80% of field samples were capable of molecular characterization. Cyst-derived DNA from FTA(®) samples can be useful for easy sampling, storage and rapid, safe and cheap shipment. The use of the FTA methodology will facilitate studies in the field to investigate the presence and genetic characterization of E. granulosus sensu lato in African countries.

  1. Pathogenicity and characterization of a novel Bacillus cereus sensu lato isolate toxic to the Mediterranean fruit fly Ceratitis capitata Wied.

    Science.gov (United States)

    Ruiu, Luca; Falchi, Giovanni; Floris, Ignazio; Marche, Maria Giovanna; Mura, Maria Elena; Satta, Alberto

    2015-03-01

    The lethal and sub-lethal effects of sporulated cultures of a novel Bacillus cereus sensu lato strain lacking detectable cry genes and identified through morphological and genetic analyses, have been studied on the Mediterranean fruit fly Ceratitis capitata. The lethal effects on young larvae were concentration dependent, with a median lethal concentration (LC50) of 4.48 × 10(8)spores/g of diet. Sporulated cultures of this strain significantly extended development time and reduced immature survival, and the size of emerging fly adults. Besides spores, the toxicity has been associated to the insoluble extra-spore fraction characterized through a proteomic approach. The profile of the extra-spore protein fraction (ES) showed major protein bands within the 35-65 kDa range. The results of mass spectrometry analysis highlighted the presence of putative virulence factors, including members of protein families previously associated to the insecticidal action of other microbial entomopathogens. These proteins include metalloproteases, peptidases and other enzymes.

  2. Deep molecular divergence and exceptional morphological stasis in dwarf cannibal snails Nata sensu lato Watson, 1934 (Rhytididae) of southern Africa.

    Science.gov (United States)

    Moussalli, Adnan; Herbert, David G

    2016-02-01

    The genus Nata Watson, 1934 is a southern African endemic belonging to the Gondwanan family of carnivorous snails, Rhytididae. We present a molecular phylogeny of the genus based on two mitochondrial (16S and COI) and two nuclear genes (ITS2 and 28S RNA), and complement this with an appraisal of morphological characters relating to both the shell and soft parts. We identify four reciprocally monophyletic lineages for which valid names are already available, plus two undescribed species restricted to the Albany Thicket Biome. We show that Nata sensu lato may not be monophyletic. Rather there exist two deep lineages within Nata s.l., one lineage potentially sister to a clade dominated by the Australian and New Zealand radiation, and the other occupying a basal position within Rhytididae. Accordingly we recommend a revision recognising two genera, namely Nata s.s. and Natella respectively. Despite deep molecular divergences within Nata s.s., phenotypic evolution has been remarkably conserved, and contrasts greatly with that exhibited across other major lineages within the Rhytididae.

  3. Molecular characterization of Echinococcus granulosus sensu lato from farm animals in Egypt.

    Directory of Open Access Journals (Sweden)

    Said Amer

    Full Text Available Little is known on the diversity and public health significance of Echinococcus species in livestock in Egypt. In this study, 37 individual hydatid cysts were collected from dromedary camels (n=28, sheep (n=7 and buffalos (n=2. DNA was extracted from protoscoleces/germinal layer of individual cysts and amplified by PCR targeting nuclear (actin II and mitochondrial (COX1 and NAD1 genes. Direct sequencing of amplicons indicated the presence of Echinococcus canadenesis (G6 genotype in 26 of 28 camel cysts, 3 of 7 sheep cysts and the 2 buffalo derived cysts. In contrast, Echinococcus granulosus sensu stricto (G1 genotype was detected in one cyst from a camel and 4 of 7 cysts from sheep, whereas Echinococcus ortleppi (G5 genotype was detected in one cyst from a camel. This is the first identification of E. ortleppi in Egypt.

  4. Molecular characterization of Echinococcus granulosus sensu lato from farm animals in Egypt.

    Science.gov (United States)

    Amer, Said; Helal, Ibrahim B; Kamau, Evelyne; Feng, Yaoyu; Xiao, Lihua

    2015-01-01

    Little is known on the diversity and public health significance of Echinococcus species in livestock in Egypt. In this study, 37 individual hydatid cysts were collected from dromedary camels (n=28), sheep (n=7) and buffalos (n=2). DNA was extracted from protoscoleces/germinal layer of individual cysts and amplified by PCR targeting nuclear (actin II) and mitochondrial (COX1 and NAD1) genes. Direct sequencing of amplicons indicated the presence of Echinococcus canadenesis (G6 genotype) in 26 of 28 camel cysts, 3 of 7 sheep cysts and the 2 buffalo derived cysts. In contrast, Echinococcus granulosus sensu stricto (G1 genotype) was detected in one cyst from a camel and 4 of 7 cysts from sheep, whereas Echinococcus ortleppi (G5 genotype) was detected in one cyst from a camel. This is the first identification of E. ortleppi in Egypt.

  5. Echinococcus granulosus sensu lato GENOTYPES IN DOMESTIC LIVESTOCK AND HUMANS IN GOLESTAN PROVINCE, IRAN.

    Science.gov (United States)

    Sharbatkhori, Mitra; Tanzifi, Asal; Rostami, Sima; Rostami, Masoomeh; Fasihi Harandi, Majid

    2016-01-01

    Cystic echinococcosis (CE) is a globally parasitic zoonosis caused by larval stages of Echinococcus granulosus. This study investigated E. granulosus genotypes isolated from livestock and humans in the Golestan province, northern Iran, southeast of the Caspian sea, using partial sequencing data of the cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) mitochondrial genes. Seventy E. granulosus isolates were collected from animals in slaughterhouses: 18 isolates from sheep, 40 from cattle, nine from camels, two from buffaloes and one from a goat, along with four human isolates (formalin-fixed, paraffin-embedded tissues) from CE patients of provincial hospitals. All isolates were successfully analysed by PCR amplification and sequencing. The sequence analysis found four E. granulosus genotypes among the 74 CE isolates: G1 (78.3%), G2 (2.7%), G3 (15%) and G6 (4%). The G1-G3 complex genotype was found in all of the sheep, goat, cattle and buffalo isolates. Among the nine camel isolates, the frequency of G1-G3 and G6 genotypes were 66.7% and 33.3%, respectively. All four human CE isolates belonged to E. granulosus sensu stricto. This study reports the first occurrence of the G2 genotype in cattle from Iran and confirms the previously reported G3 genotype in camels in the same country.

  6. Bayesian inference towards the resolution of molecular evolution:application to the "Trichoderma harzianum sensu lato" clade

    Institute of Scientific and Technical Information of China (English)

    Druzhinina I S; Kubicek C P

    2004-01-01

    or T. harzianum formed either a proliferating megaclade (three loci combined tree) or appeared as several independent branches on the star-like phylograrm (ITS1 and 2 and large tef1a intron) . However most terminal clacles had a high posterior probability and thus fulfilled the criterion to consider their member isolates as a phylogenetic species. The clade containing the ex-neotype strain of T. harzianum included exclusively clonally propagating isolates from temperate climates, and was clearly distinguished from all other "H. lixii/T. harzianum" lineages by multivariate analysis of phenotype microarray data. We propose that only this lineage should be regarded as T. harzianum sensu stricto, and that the other lineages be regarded as T.harzianum sensu lato until phenetic characters have been defined which allow their description as separate taxa.

  7. 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.

  8. Seasonal Variation in Biting Rates of Simulium damnosum sensu lato, Vector of Onchocerca volvulus, in Two Sudanese Foci.

    Directory of Open Access Journals (Sweden)

    Isam M A Zarroug

    Full Text Available The abundance of onchocerciasis vectors affects the epidemiology of disease in Sudan, therefore, studies of vector dynamics are crucial for onchocerciasis control/elimination programs. This study aims to compare the relative abundance, monthly biting-rates (MBR and hourly-based distribution of onchocerciasis vectors in Abu-Hamed and Galabat foci. These seasonally-based factors can be used to structure vector control efforts to reduce fly-biting rates as a component of onchocerciasis elimination programs.A cross-sectional study was conducted in four endemic villages in Abu-Hamed and Galabat foci during two non-consecutive years (2007-2008 and 2009-2010. Both adults and aquatic stages of the potential onchocerciasis vector Simulium damnosum sensu lato were collected following standard procedures during wet and dry seasons. Adult flies were collected using human landing capture for 5 days/month. The data was recorded on handheld data collection sheets to calculate the relative abundance, MBR, and hourly-based distribution associated with climatic factors. The data analysis was carried out using ANOVA and Spearman rank correlation tests.Data on vector surveillance revealed higher relative abundance of S. damnosum s.l. in Abu- Hamed (39,934 flies than Galabat (8,202 flies. In Abu-Hamed, vector populations increased in January-April then declined in June-July until they disappeared in August-October. Highest black fly density and MBR were found in March 2007 (N = 9,444, MBR = 58,552.8 bites/person/month, and March 2010 (N = 2,603, MBR = 16,138.6 bites/person/month while none of flies were collected in August-October (MBR = 0 bites/person/month. In Galabat, vectors increased in September-December, then decreased in February-June. The highest vector density and MBR were recorded in September 2007 (N = 1,138, MBR = 6,828 bites/person/month and September 2010 (N = 1,163, MBR = 6,978 bites/person/month, whereas, none appeared in collection from April to

  9. A novel PCR-RFLP assay for molecular characterization of Echinococcus granulosus sensu lato and closely related species in developing countries.

    Science.gov (United States)

    Chaâbane-Banaoues, Raja; Oudni-M'rad, Myriam; M'rad, Selim; Amani, Hizem; Mezhoud, Habib; Babba, Hamouda

    2016-10-01

    Cystic echinococcosis, due to Echinococcus granulosus sensu lato (s. l.), currently affects three million people, especially in low-income countries and results in high livestock production loss. DNA-based methods demonstrated genetic variability of E. granulosus s. l., and five species were recognized to belong to the complex, including E. granulosus sensu stricto (s.s) (genotypes G1-G3), Echinococcus equinus (genotype G4), Echinococcus ortleppi (genotype G5), Echinococcus canadensis (genotypes G6-G10), and the lion strain Echinococcus felidis. The characterization of Echinococcus species responsible for human and animal echinococcosis is crucial to adapt the preventive measures against this parasitic disease. The sequencing approach is the gold standard for genotyping assays. Unfortunately, developing countries do not often have access to these techniques. Based on in silico RFLP tools, we described an accurate PCR-RFLP method for Echinococcus spp. characterization. The double digestion with the HaeIII and HinfI restriction enzymes of the PCR product from nad1 gene (1071 bp) led to a clear discrimination between E. granulosus s. l. and most closely related species (Echinococcus shiquicus and Echinococcus multilocularis).Molecular procedures and phylogenetic analysis confirmed the efficiency and the reproducibility of this simple and fast PCR-RFLP method. This technique is proved useful for fresh/unfixed and FF-PET tissues and enables large-scale molecular epidemiological screening in developing countries.

  10. 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.

  11. 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....... burgdorferi sensu stricto B. afzelii, B. garinii, B. valaisiana, B. lusitaniae in nymphs, by IFA and PCR. 600 nymphs were collected in North Zealand of Denmark. Each nymph was first analysed by IFA. If positive for spirochaetal infection, the genospecies was determined by PCR. The infection rate of B....... burgdorferi sensu lato was 15.5%, with the primary genospecies being B. afzelii (64.3%), B. garinii (57.1%), and B. lusitaniae (26.8%). It is the first time B. lusitaniae is documented in Denmark. Even though, the highest infection rate was discovered for B. afzelii and B. garinii, mixed infections are more...

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

    OpenAIRE

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

    2013-01-01

    Borrelia (B.) burgdorferi sensu lato, the causative agent of Lyme disease, is the most important arthropodborne 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 t...

  13. Comparative molecular analyses of Borrelia burgdorferi sensu stricto strains B31 and N40D10/E9 and determination of their pathogenicity

    Directory of Open Access Journals (Sweden)

    Chan Kamfai

    2012-07-01

    Full Text Available Abstract Background Lyme disease in the United States is caused primarily by B. burgdorferi sensu stricto while other species are also prevalent in Europe. Genetic techniques have identified several chromosomal and plasmid-borne regulatory and virulence factors involved in Lyme pathogenesis. B31 and N40 are two widely studied strains of B. burgdorferi, which belong to two different 16 S-23 S rRNA spacer types (RST and outer surface protein C (OspC allelic groups. However, the presence of several known virulence factors in N40 has not been investigated. This is the first comprehensive study that compared these two strains both in vitro and using the mouse model of infection. Results Phylogenetic analyses predict B31 to be more infectious. However, our studies here indicate that N40D10/E9 is more infectious than the B31 strain at lower doses of inoculation in the susceptible C3H mice. Based-upon a careful analyses of known adhesins of these strains, it is predicted that the absence of a known fibronectin-glycosaminoglycan binding adhesin, bbk32, in the N40 strain could at least partially be responsible for reduction in its binding to Vero cells in vitro. Nevertheless, this difference does not affect the infectivity of N40D10/E9 strain. The genes encoding known regulatory and virulence factors critical for pathogenesis were detected in both strains. Differences in the protein profiles of these B. burgdorferi strains in vitro suggest that the novel, differentially expressed molecules may affect infectivity of B. burgdorferi. Further exacerbation of these molecular differences in vivo could affect the pathogenesis of spirochete strains. Conclusion Based upon the studies here, it can be predicted that N40D10/E9 disseminated infection at lower doses may be enhanced by its lower binding to epithelial cells at the site of inoculation due to the absence of BBK32. We suggest that complete molecular analyses of virulence factors followed by their evaluation

  14. Infection with Ehrlichia canis and Anaplasma platys (Rickettsiales: Anaplasmataceae) in two lineages of Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) from Argentina.

    Science.gov (United States)

    Cicuttin, Gabriel L; Tarragona, Evelina L; De Salvo, M Nazarena; Mangold, Atilio J; Nava, Santiago

    2015-09-01

    Natural infection with Ehrlichia canis and Anaplasma platys in ticks belonging to the tropical and temperate lineages of Rhipicephalus sanguineus sensu lato from Argentina was evaluated. Samples were tested for Ehrlichia canis infection by PCR assays using 16S rRNA, dsb and p28 gene, while detection of A. platys was performed with 16S rRNA and groESL gene. The assignment of the ticks to each lineage was corroborated with 16S rDNA sequences. All ticks infected with E. canis and A. platys belonged to the tropical lineage. These results constitute the first record of E. canis infection in R. sanguineus s.l ticks from Argentina. No ticks from the temperate lineage were found to be infected with E. canis, coinciding with previous studies performed in Argentina and Uruguay where E. canis infection was not detected in R. sanguineus s.l from the temperate lineage. Because the presence of the tropical lineage of R. sanguineus s.l has been documented in tropical areas of northern Argentina between 22° and 24° of south latitude, the findings of this work indicate that transmission of E. canis and A. platys to dogs by R. sanguineus s.l probably occurs along this region.

  15. Seroprevalence of Rickettsia spp. in Equids and Molecular Detection of 'Candidatus Rickettsia amblyommii' in Amblyomma cajennense Sensu Lato Ticks From the Pantanal Region of Mato Grosso, Brazil.

    Science.gov (United States)

    Alves, Alvair Da S; Melo, Andréia L T; Amorim, Marcus V; Borges, Alice M C M; Gaíva E Silva, Lucas; Martins, Thiago F; Labruna, Marcelo B; Aguiar, Daniel M; Pacheco, Richard C

    2014-11-01

    The aim of the study was to evaluate exposure of equids to rickettsial agents (Rickettsia rickettsii, Rickettsia parkeri, 'Candidatus Rickettsia amblyommii', Rickettsia rhipicephali, and Rickettsia bellii) and rickettsial infection in ticks of a Pantanal region of Brazil. Sera of 547 equids (500 horses and 47 donkeys) were evaluated by indirect immunofluorescence assay. In total, 665 adults and 106 nymphal pools of Amblyomma cajennense F. sensu lato, 10 Dermacentor nitens Neumann ticks, and 88 larval pools of Amblyomma sp. were tested by polymerase chain reaction (PCR). Overall, 337 (61.6%) equids were reactive (titer ≥64) to at least one antigen of Rickettsia spp. The prevalence values for Rickettsia were 66%, and the highest endpoint titers were observed for 'Ca. R. amblyommii'. By PCR 3 (0.45%) A. cajennense s.l. females were positive for 'Ca. R. amblyommii'. Minimum infection rates of 0.75% for nymphs and 0.34% for larvae were calculated. Positive samples of ticks have had a fragment of the 16S mitochondrial rRNA gene sequenced and sequences showed 99% identity to Amblyomma sculptum Berlese. This study reports a wide exposure of equids to Rickettsia agents, and PCR evidence of infection with 'Ca. R. amblyommii', for the first time, in A. sculptum.

  16. Morphological characteristics of the antennal flagellum and its sensilla chaetica with character displacement in the sandfly Phlebotomus argentipes Annandale and Brunetti sensu lato (Diptera: Psychodidae)

    Indian Academy of Sciences (India)

    K Ilango

    2000-06-01

    Using light microscope and scanning electron microscope, the external morphological characteristics of the antennal flagellum and its sensilla are described in the sandfly, Phlebotomus argentipes Annandale and Brunetti sensu lato, a well known vector of visceral leishmaniasis in India. A revised terminology is given for the antennal segments to bring phlebotomine more in line with other subfamilies and families while a description of antennal sensilla is provided for the first time in phlebotomine sandflies. Each flagellum consists of scape, pedicel, flagellomeres I to XIII and apiculus. The antennal segments contain scales and sensilla and the latter consist of sensilla trichodea, s. basiconica, s. auricillica, s. coeloconica and s. chaetica and their putative functions are discussed. The sensilla chaeticum hitherto known as antennal ascoid in the phlebotomine sandflies was used to differentiate within and between species. Differences in its relative size to the flagellomere between the populations of P. argentipes collected from the endemic and non-endemic areas in Tamil Nadu state, southern India were established. These differences are considered to be a character displacement as means of premating reproductive isolating mechanism among the populations/members of species complex.

  17. Fitness costs to Helicoverpa armigera after exposure to sub-lethal concentrations of Metarhizium anisopliae sensu lato: Study on F1 generation.

    Science.gov (United States)

    Jarrahi, Azadeh; Safavi, Seyed Ali

    2016-07-01

    The entomopathogenic fungus, Metarhizium anisopliae (Metsch.) Sorokin is a valuable biocontrol agent attacking larval stages of many lepidopteran pests including Helicoverpa armigera (Hübner). Sub-lethal effects of M. anisopliae sensu lato (s.l.) (isolate M14) were investigated on life table parameters of offspring from treated larvae of H. armigera. Duration of different life stages was significantly affected by fungal treatments. Fecundity was decreased in females derived from H. armigera larvae treated with M. anisopliae s.l. Sub-lethal concentrations of the entomopathogen reduced the net reproduction rate (R0) of F1 insects for all treatments compared with the control. Similar reductions were observed for the intrinsic and the finite rates of increase (rm and λ, respectively). The mean generation time (T) and the doubling time (DT) were statistically higher in offspring of individuals exposed to some fungal concentrations than control insects. Our results indicated that there was a significant decrease in the F1 population of H. armigera derived from larvae that were exposed to sub-lethal concentrations of M. anisopliae s.l.

  18. Utility of COI, CAD and morphological data for resolving relationships within the genus Sarcophaga (sensu lato) (Diptera: Sarcophagidae): a preliminary study.

    Science.gov (United States)

    Meiklejohn, Kelly A; Wallman, James F; Pape, Thomas; Cameron, Stephen L; Dowton, Mark

    2013-10-01

    Currently there are ≈ 3000 known species of Sarcophagidae (Diptera), which are classified into 173 genera in three subfamilies. Almost 25% of sarcophagids belong to the genus Sarcophaga (sensu lato) however little is known about the validity of, and relationships between the ≈ 150 (or more) subgenera of Sarcophaga s.l. In this preliminary study, we evaluated the usefulness of three sources of data for resolving relationships between 35 species from 14 Sarcophaga s.l. subgenera: the mitochondrial COI barcode region, ≈ 800 bp of the nuclear gene CAD, and 110 morphological characters. Bayesian, maximum likelihood (ML) and maximum parsimony (MP) analyses were performed on the combined dataset. Much of the tree was only supported by the Bayesian and ML analyses, with the MP tree poorly resolved. The genus Sarcophaga s.l. was resolved as monophyletic in both the Bayesian and ML analyses and strong support was obtained at the species-level. Notably, the only subgenus consistently resolved as monophyletic was Liopygia. The monophyly of and relationships between the remaining Sarcophaga s.l. subgenera sampled remain questionable. We suggest that future phylogenetic studies on the genus Sarcophaga s.l. use combined datasets for analyses. We also advocate the use of additional data and a range of inference strategies to assist with resolving relationships within Sarcophaga s.l.

  19. Geographical distribution and prevalence of Borrelia burgdorferi genospecies in questing Ixodes ricinus from Romania: a countrywide study.

    Science.gov (United States)

    Kalmár, Zsuzsa; Mihalca, Andrei D; Dumitrache, Mirabela O; Gherman, Călin M; Magdaş, Cristian; Mircean, Viorica; Oltean, Miruna; Domşa, Cristian; Matei, Ioana A; Mărcuţan, Daniel I; Sándor, Attila D; D'Amico, Gianluca; Paştiu, Anamaria; Györke, Adriana; Gavrea, Raluca; Marosi, Béla; Ionică, Angela; Burkhardt, Etelka; Toriay, Hortenzia; Cozma, Vasile

    2013-09-01

    The paper reports the prevalence and geographical distribution of Borrelia burgdorferi sensu lato (s.l.) and its genospecies in 12,221 questing Ixodes ricinus ticks collected at 183 locations from all the 41 counties of Romania. The unfed ticks were examined for the presence of B. burgdorferi s.l. by PCR targeting the intergenic spacer 5S-23S. Reverse line blot hybridization (RLB) and restriction fragment length polymorphism (RFLP) analysis were performed for identification of B. burgdorferi genospecies. The overall prevalence of infection was 1.4%, with an average local prevalence between 0.75% and 18.8%. B. burgdorferi s.l. was found in ticks of 55 of the 183 localities. The overall prevalence B. burgdorferi s.l. in ticks in the infected localities was 3.8%. The total infection prevalence was higher in female ticks than in other developmental stages. Three Borrelia genospecies were detected. The most widely distributed genospecies was B. afzelii, followed by B. garinii and B. burgdorferi sensu stricto (s.s.). The study is the first countrywide study and the first report of B. burgdorferi s.s. in Romania. The distribution maps show that higher prevalences were recorded in hilly areas, but Lyme borreliosis spirochetes were also present in forested lowlands, albeit with a lower prevalence.

  20. 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.

  1. Investigating the host-range of the rust fungus Puccinia psidii sensu lato across tribes of the family Myrtaceae present in Australia.

    Science.gov (United States)

    Morin, Louise; Aveyard, Ruth; Lidbetter, Jonathan R; Wilson, Peter G

    2012-01-01

    The exotic rust fungus Puccinia psidii sensu lato was first detected in Australia in April 2010. This study aimed to determine the host-range potential of this accession of the rust by testing its pathogenicity on plants of 122 taxa, representative of the 15 tribes of the subfamily Myrtoideae in the family Myrtaceae. Each taxon was tested in two separate trials (unless indicated otherwise) that comprised up to five replicates per taxon and six replicates of a positive control (Syzygium jambos). No visible symptoms were observed on the following four taxa in either trial: Eucalyptus grandis×camaldulensis, E. moluccana, Lophostemon confertus and Sannantha angusta. Only small chlorotic or necrotic flecks without any uredinia (rust fruiting bodies) were observed on inoculated leaves of seven other taxa (Acca sellowiana, Corymbia calophylla 'Rosea', Lophostemon suaveolens, Psidium cattleyanum, P. guajava 'Hawaiian' and 'Indian', Syzygium unipunctatum). Fully-developed uredinia were observed on all replicates across both trials of 28 taxa from 8 tribes belonging to the following 17 genera: Agonis, Austromyrtus, Beaufortia, Callistemon, Calothamnus, Chamelaucium, Darwinia, Eucalyptus, Gossia, Kunzea, Leptospermum, Melaleuca, Metrosideros, Syzygium, Thryptomene, Tristania, Verticordia. In contrast, the remaining 83 taxa inoculated, including the majority of Corymbia and Eucalyptus species, developed a broad range of symptoms, often across the full spectrum, from fully-developed uredinia to no visible symptoms. These results were encouraging as they indicate that some levels of genetic resistance to the rust possibly exist in these taxa. Overall, our results indicated no apparent association between the presence or absence of disease symptoms and the phylogenetic relatedness of taxa. It is most likely that the majority of the thousands of Myrtaceae species found in Australia have the potential to become infected to some degree by the rust, although this wide host range may

  2. Investigating the host-range of the rust fungus Puccinia psidii sensu lato across tribes of the family Myrtaceae present in Australia.

    Directory of Open Access Journals (Sweden)

    Louise Morin

    Full Text Available The exotic rust fungus Puccinia psidii sensu lato was first detected in Australia in April 2010. This study aimed to determine the host-range potential of this accession of the rust by testing its pathogenicity on plants of 122 taxa, representative of the 15 tribes of the subfamily Myrtoideae in the family Myrtaceae. Each taxon was tested in two separate trials (unless indicated otherwise that comprised up to five replicates per taxon and six replicates of a positive control (Syzygium jambos. No visible symptoms were observed on the following four taxa in either trial: Eucalyptus grandis×camaldulensis, E. moluccana, Lophostemon confertus and Sannantha angusta. Only small chlorotic or necrotic flecks without any uredinia (rust fruiting bodies were observed on inoculated leaves of seven other taxa (Acca sellowiana, Corymbia calophylla 'Rosea', Lophostemon suaveolens, Psidium cattleyanum, P. guajava 'Hawaiian' and 'Indian', Syzygium unipunctatum. Fully-developed uredinia were observed on all replicates across both trials of 28 taxa from 8 tribes belonging to the following 17 genera: Agonis, Austromyrtus, Beaufortia, Callistemon, Calothamnus, Chamelaucium, Darwinia, Eucalyptus, Gossia, Kunzea, Leptospermum, Melaleuca, Metrosideros, Syzygium, Thryptomene, Tristania, Verticordia. In contrast, the remaining 83 taxa inoculated, including the majority of Corymbia and Eucalyptus species, developed a broad range of symptoms, often across the full spectrum, from fully-developed uredinia to no visible symptoms. These results were encouraging as they indicate that some levels of genetic resistance to the rust possibly exist in these taxa. Overall, our results indicated no apparent association between the presence or absence of disease symptoms and the phylogenetic relatedness of taxa. It is most likely that the majority of the thousands of Myrtaceae species found in Australia have the potential to become infected to some degree by the rust, although this

  3. Combining a climatic niche model of an invasive fungus with its host species distributions to identify risks to natural assets: Puccinia psidii Sensu Lato in Australia.

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    Darren J Kriticos

    Full Text Available Puccinia psidii sensu lato (s.l. is an invasive rust fungus threatening a wide range of plant species in the family Myrtaceae. Originating from Central and South America, it has invaded mainland USA and Hawai'i, parts of Asia and Australia. We used CLIMEX to develop a semi-mechanistic global climatic niche model based on new data on the distribution and biology of P. psidii s.l. The model was validated using independent distribution data from recently invaded areas in Australia, China and Japan. We combined this model with distribution data of its potential Myrtaceae host plant species present in Australia to identify areas and ecosystems most at risk. Myrtaceaeous species richness, threatened Myrtaceae and eucalypt plantations within the climatically suitable envelope for P. psidii s.l in Australia were mapped. Globally the model identifies climatically suitable areas for P. psidii s.l. throughout the wet tropics and sub-tropics where moist conditions with moderate temperatures prevail, and also into some cool regions with a mild Mediterranean climate. In Australia, the map of species richness of Myrtaceae within the P. psidii s.l. climatic envelope shows areas where epidemics are hypothetically more likely to be frequent and severe. These hotspots for epidemics are along the eastern coast of New South Wales, including the Sydney Basin, in the Brisbane and Cairns areas in Queensland, and in the coastal region from the south of Bunbury to Esperance in Western Australia. This new climatic niche model for P. psidii s.l. indicates a higher degree of cold tolerance; and hence a potential range that extends into higher altitudes and latitudes than has been indicated previously. The methods demonstrated here provide some insight into the impacts an invasive species might have within its climatically suited range, and can help inform biosecurity policies regarding the management of its spread and protection of valued threatened assets.

  4. Comparative efficacy of two oral treatments for dogs containing either afoxolaner or fluralaner against Rhipicephalus sanguineus sensu lato and Dermacentor reticulatus.

    Science.gov (United States)

    Beugnet, Frederic; Liebenberg, Julian; Halos, Lenaïg

    2015-04-15

    The present study compares the efficacy of two recent oral ectoparasiticides containing isoxazolines (NexGard(®), containing afoxolaner and administered at a monthly regimen, and Bravecto™ containing fluralaner and administered at a tri-monthly regimen) against Rhipicephalus sanguineus sensu lato and Dermacentor reticulatus ticks on dogs. 24 dogs were randomly allocated to untreated control, NexGard(®) treated, and Bravecto™ treated groups. The treatments were administered on Days 0, 28 and 56 for afoxolaner and on Day 0 for fluralaner. Tick infestations were performed weekly with 50 unfed adult ticks per each species on each dog from Days 30 to 84 (with the exception of R. sanguineus on Day 63). Ticks were counted at 24h post-infestation. The dogs from both treated groups had statistically significantly (p<0.05) less R. sanguineus and D. reticulatus ticks compared to the untreated dogs on all assessment days. Percent efficacy against R. sanguineus ranged from 86.4% to 99.5% at 24h post-infestation for NexGard(®) and from 65.7% to 100% for Bravecto™. Statistically significantly (p<0.05) less R. sanguineus ticks were recorded for NexGard(®) treated dogs compared to Bravecto™ treated dogs on Day 78. Percent efficacy against D. reticulatus ranged from 85.2% to 99.6% at 24h post-infestation for NexGard(®) and from 63.4% to 99.1% for Bravecto™. Statistically significantly (p<0.05) less D. reticulatus ticks were recorded for NexGard(®) treated dogs compared to Bravecto™ treated dogs on Days 71, 78 and 85.

  5. Investigation on resources of Cordyceps sensu lato in Wanfoshan reserve %安徽省万佛山地区虫草资源的初步调查

    Institute of Scientific and Technical Information of China (English)

    彭凡; 曹珊; 张琪; 李春如; 樊美珍

    2011-01-01

    通过对万佛山自然保护区的多次野外采集和调查,研究发现万佛山地区分布的虫草分属.6个种,分别为丝虫草Ophiocordyceps filiformis、珊瑚虫草Cordyceps martialis、粉被虫草C.pruinosa、下垂虫草O.nutans、亚黄蜂虫草O.oxycephala和莲状虫草C.nelumboides.其中下垂虫草和丝虫草最多,分别占总数的28%和23%.虫草的寄主涉及到鳞翅目的幼虫和茧、蜂的成虫、蜘蛛和蝽象,而主要寄主为鳞翅目的幼虫,占总数的58%.莲状虫草为中国大陆新记录种,此前仅在日本和中国台湾省有过报道.%Six species of Cordyceps sensu lato were collected from Wanfoshan Nature Reserve. They were Ophiocordyceps filiformis Moureau, Cordyceps martialis Spegazzini, C. pruinosa Petch, O. nutans Pat., O. oxycephala Penz. & Sacc. and C. nelumboides. Kobayasi & Shimizu. O. nutans and O. filiformis were the dominant species with relative abundance of 28% and 23%. The host related to lepodopteran larvae and cocoon, adult of bee,arachnida and pentatomidae, the main hosts were lepodopteran larvae with relative abundance of 58%. C. nelumboides was a new record in Chinese mainland, and it had only been reported in Japan and Taiwan Province before.

  6. Development of a powder formulation based on Bacillus cereus sensu lato strain B25 spores for biological control of Fusarium verticillioides in maize plants.

    Science.gov (United States)

    Martínez-Álvarez, Juan C; Castro-Martínez, Claudia; Sánchez-Peña, Pedro; Gutiérrez-Dorado, Roberto; Maldonado-Mendoza, Ignacio E

    2016-05-01

    Maize is an economically important crop in northern Mexico. Different fungi cause ear and root rot in maize, including Fusarium verticillioides (Sacc.) Nirenberg. Crop management of this pathogen with chemical fungicides has been difficult. By contrast, the recent use of novel biocontrol strategies, such as seed bacterization with Bacillus cereus sensu lato strain B25, has been effective in field trials. These approaches are not without their problems, since insufficient formulation technology, between other factors, can limit success of biocontrol agents. In response to these drawbacks, we have developed a powder formulation based on Bacillus B25 spores and evaluated some of its characteristics, including shelf life and efficacy against F. verticillioides, in vitro and in maize plants. A talc-based powder formulation containing 1 × 10(9) c.f.u. g(-1) was obtained and evaluated for seed adherence ability, seed germination effect, shelf life and antagonism against F. verticillioides in in vitro and in planta assays. Seed adherence of viable bacterial spores ranged from 1.0 to 1.41 × 10(7) c.f.u. g(-1). Bacteria did not display negative effects on seed germination. Spore viability for the powder formulation slowly decreased over time, and was 53 % after 360 days of storage at room temperature. This formulation was capable of controlling F. verticillioides in greenhouse assays, as well as eight other maize phytopathogenic fungi in vitro. The results suggest that a talc-based powder formulation of Bacillus B25 spores may be sufficient to produce inoculum for biocontrol of maize ear and root rots caused by F. verticillioides.

  7. Outer Surface Protein C Peptide Derived from Borrelia burgdorferi Sensu Stricto as a Target for Serodiagnosis of Early Lyme Disease

    Science.gov (United States)

    Arnaboldi, Paul M.; Seedarnee, Rudra; Sambir, Mariya; Callister, Steven M.; Imparato, Josephine A.

    2013-01-01

    Current serodiagnostic assays for Lyme disease are inadequate at detecting early infection due to poor sensitivity and nonspecificity that arise from the use of whole bacteria or bacterial proteins as assay targets; both targets contain epitopes that are cross-reactive with epitopes found in antigens of other bacterial species. Tests utilizing peptides that contain individual epitopes highly specific for Borrelia burgdorferi as diagnostic targets are an attractive alternative to current assays. Using an overlapping peptide library, we mapped linear epitopes in OspC, a critical virulence factor of B. burgdorferi required for mammalian infection, and confirmed the results by enzyme-linked immunosorbent assay (ELISA). We identified a highly conserved 20-amino-acid peptide epitope, OspC1. Via ELISA, OspC1 detected specific IgM and/or IgG in 60 of 98 serum samples (62.1%) obtained from patients with erythema migrans (early Lyme disease) at the time of their initial presentation. By comparison, the commercially available OspC peptide PepC10 detected antibody in only 48 of 98 serum samples (49.0%). In addition, OspC1 generated fewer false-positive results among negative healthy and diseased (rheumatoid arthritis and positive Rapid Plasma Reagin [RPR+] test result) control populations than did PepC10. Both highly specific and more sensitive than currently available OspC peptides, OspC1 could have value as a component of a multipeptide Lyme disease serological assay with significantly improved capabilities for the diagnosis of early infection. PMID:23365204

  8. Multilocus sequence typing of Pseudomonas syringae sensu lato confirms previously described genomospecies and permits rapid identification of P. syringae pv. coriandricola and P. syringae pv. apii causing bacterial leaf spot on parsley.

    Science.gov (United States)

    Bull, Carolee T; Clarke, Christopher R; Cai, Rongman; Vinatzer, Boris A; Jardini, Teresa M; Koike, Steven T

    2011-07-01

    Since 2002, severe leaf spotting on parsley (Petroselinum crispum) has occurred in Monterey County, CA. Either of two different pathovars of Pseudomonas syringae sensu lato were isolated from diseased leaves from eight distinct outbreaks and once from the same outbreak. Fragment analysis of DNA amplified between repetitive sequence polymerase chain reaction; 16S rDNA sequence analysis; and biochemical, physiological, and host range tests identified the pathogens as Pseudomonas syringae pv. apii and P. syringae pv. coriandricola. Koch's postulates were completed for the isolates from parsley, and host range tests with parsley isolates and pathotype strains demonstrated that P. syringae pv. apii and P. syringae pv. coriandricola cause leaf spot diseases on parsley, celery, and coriander or cilantro. In a multilocus sequence typing (MLST) approach, four housekeeping gene fragments were sequenced from 10 strains isolated from parsley and 56 pathotype strains of P. syringae. Allele sequences were uploaded to the Plant-Associated Microbes Database and a phylogenetic tree was built based on concatenated sequences. Tree topology directly corresponded to P. syringae genomospecies and P. syringae pv. apii was allocated appropriately to genomospecies 3. This is the first demonstration that MLST can accurately allocate new pathogens directly to P. syringae sensu lato genomospecies. According to MLST, P. syringae pv. coriandricola is a member of genomospecies 9, P. cannabina. In a blind test, both P. syringae pv. coriandricola and P. syringae pv. apii isolates from parsley were correctly identified to pathovar. In both cases, MLST described diversity within each pathovar that was previously unknown.

  9. Mechanisms of karyotype differentiation in Cassidinae sensu lato (Coleoptera, Polyphaga, Chrysomelidae) based on seven species of the Brazilian fauna and an overview of the cytogenetic data.

    Science.gov (United States)

    de Julio, Milena; Fernandes, Flávia Rodrigues; Costa, Cleide; Almeida, Mara Cristina; Cella, Doralice Maria

    2010-01-01

    Among the subfamilies of Chrysomelidae, Cassidinae sensu lato (s.l.) includes 6000 species distributed in 43 tribes. Approximately 100 of these species were cytogenetically analyzed and most of them presented 2n=18=16+Xy(p), which was smaller than 2n=20=18+Xy(p) considered basal for Polyphaga. However, some groups of species presented maintenance of the basal diploid number and others showed increase in this number. Certain species of the latter group also exhibited variation in the type of sex chromosome system (SCS). Considering the recent taxonomic revision accomplished for the Cassidinae s.l. species, the existence of phylogenetic relationship for some species of this subfamily, the high diversity of species of this group in the Neotropical region, and the low number of Cassidinae s.l. species karyotyped so far, the aim of the present work was to establish the main mechanisms involved in the karyotype evolution of this subfamily through the study of seven species of the Brazilian fauna and overview of the cytogenetic data. The individuals were collected in southeast and south of Brazil. The chromosomal preparations obtained from embryo and testes of adult males were stained with Giemsa solution. The species Agroiconota inedita (2n=42=40+Xy(p)), Charidotella (s.str.) immaculata (2n=22=20+Xy(p)), Charidotella (s.str.) sexpunctata (2n=22=20+Xy(p)), and Stolas chalybaea (2n=24=22+Xy(p)) revealed diploid number higher than that established as basal for Polyphaga and biarmed chromosomes. The karyotype of Cteisella confusa, Deloyala cruciata, and Metriona elatior showed the chromosomal formulae 2n=18=16+Xy(p) considered modal for Cassidinae s.l. and biarmed chromosomes. The seven species exhibited easily identified sex chromosomes due to their size and/or morphology. The analysis of meiotic cells of all the species showed pachytenes with a positively heteropycnotic block probably corresponding to the sex chromosomes; diplotenes with a high number of bivalents with two

  10. First record of Borrelia burgdorferi B31 strain in Dermacentor nitens ticks in the northern region of Parana (Brazil

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    Daniela Dib Gonçalves

    2013-09-01

    Full Text Available The aim of this study was to investigate the presence of DNA of Borrelia burgdorferi sensu lato (s.l. in ticks that feed on horses used for animal traction in rural Jataizinho, Parana, Brazil. Between February and June 2008, a total of 224 ticks was collected of which 75% were identified as Dermacentor nitens and 25% as Amblyomma cajenense. To amplify B. burgdorferi s.l. DNA, the intergenic space region (ISR between the 5S (rrf 23S (rrl rRNA genes was used as targets for nested-PCR. Two ticks of the D. nitens species were positive for B. burgdorferi s.l. Both species showed a fragment of 184 bp, but the sequencing revealed 99.9% homology with the B. burgdorferi sensu stricto (s.s. strain B31. These results showed, for the first time, the presence of spirochete DNA infecting ticks that parasitize horses used for animal traction, in the rural municipality mentioned. In conclusion, this study opens up promising prospects for determining the infection rate of B. burgdorferi s.s. genospecies or other species in the equine population, as well as the impact of the infection rate on Lyme disease in the state of Parana.

  11. Reassessment of the taxonomic structure of the diazotrophic genus Azoarcus sensu lato and description of three new genera and new species, Azovibrio restrictus gen. nov., sp. nov., Azospira oryzae gen. nov., sp. nov. and Azonexus fungiphilus gen. nov., sp. nov.

    Science.gov (United States)

    Reinhold-Hurek, B; Hurek, T

    2000-03-01

    The taxonomic structure of members of the genus Azoarcus sensu lato was reassessed in a polyphasic approach. Two species, Azoarcus communis and Azoarcus indigens, three unnamed species containing diazotrophs associated with Kallar grass roots (groups C, D) and a group of strains (E) isolated from fungi were analysed. They were compared by PAGE analyses of cellular proteins, genomic fingerprints, morphological and nutritional features to new isolates from rice roots. All strains within groups C, D and E containing 5-12 isolates showed group-specific cell and colony morphology and carbon source utilization patterns, with exception of the obligately microaerobic strain BS20-3, a member of group C. All strains, with this exception, also had almost indistinguishable electrophoretic protein patterns and genomic fingerprints generated with tDNA-directed primers, suggesting they belong to the same species. Phylogenetic analyses of almost complete 16S rDNA sequences carried out with three different algorithms (neighbour-joining, maximum-likelihood, parsimony) revealed that Azoarcus sensu lato is not monophyletic. Groups C, D and E formed three distinct lineages located between the Azoarcus/Thauera and the Rhodocyclus clusters. Phylogenetic distances between groups C, D and E were as large as between other genera (93-94% sequence similarity). This suggested they have the rank of three different genera. Since it was possible to differentiate them from each other and other related bacteria by phenotypic features, three new genera with one type species each are proposed: Azovibrio restrictus gen. nov., sp. nov., Azospira oryzae gen. nov., sp. nov. and Azonexus fungiphilus gen. nov., sp. nov.

  12. Prevalence of serological response to Borrelia burgdorferi in farmers from eastern and central Poland.

    Science.gov (United States)

    Zając, V; Pinkas, J; Wójcik-Fatla, A; Dutkiewicz, J; Owoc, A; Bojar, I

    2017-03-01

    Lyme borreliosis (Lyme disease) caused by the Borrelia burgdorferi sensu lato spirochete is the most common tick-borne infection manifested by a wide spectrum of clinical symptoms. In Poland, the preventive health care does not comprise individual farmers as it is practiced in foresters. The objective of this study was to evaluate the exposure of Polish farmers to infection with B. burgdorferi, based on serological screening test and epidemiological investigation. A total of 3,597 farmers were examined for the presence of B. burgdorferi antibodies, as well as interviewed regarding exposure to ticks and prophylaxis of tick-borne diseases. The prevalence varied between 18.2 and 50.7 % suggesting a focal occurrence of borreliosis. A significant increase in the frequency of positive reactions in the oldest age ranges was observed, equaling 30.9 % in the range of 60-69 years and 53.6 % in the range of 80-91 years. The prevalence of the anti-B. burgdorferi antibodies of IgG class (14.7 %) was similar to that of IgM class (16.0 %). Seroreactivity to B. burgdorferi antigen was significantly higher in the group of farmers exposed to repeated tick bites. Significant relationships were also found between some other risk factors and occurrence of seropositive reactions to B. burgdorferi. To the best of our knowledge, this is the first study concerning seroprevalence to B. burgdorferi carried out on such a large group of farmers. Results indicate a high risk of B. burgdorferi infection among Polish farmers and associations between some risk factors and the presence of seropositive reactions.

  13. 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.

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

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    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.

  15. 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.

  16. 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

  17. Polymorphism of 41 kD Flagellin Gene and Its Human B-Cell Epitope in Borrelia burgdorferi Strains of China

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    Huixin Liu

    2016-01-01

    Full Text Available The 41 kD flagellin of Borrelia burgdorferi (B. burgdorferi is a major component of periplasmic flagellar filament core and a good candidate for serodiagnosis in early stage of Lyme disease. Here, we chose 89 B. burgdorferi strains in China, amplified the gene encoding the 41 kD flagellin, and compared the sequences. The results showed that genetic diversity presented in the 41 kD flagellin genes of all 89 strains among the four genotypes of B. burgdorferi, especially in the genotype of B. garinii. Some specific mutation sites for each genotype of the 41 kD flagellin genes were found, which could be used for genotyping B. burgdorferi strains in China. Human B-cell epitope analysis showed that thirteen of 15 nonsynonymous mutations occurred in the epitope region of 41 kD flagellin and thirty of 42 B-cell epitopes were altered due to all 13 nonsynonymous mutations in the epitope region, which may affect the function of the antigen. Nonsynonymous mutations and changed human B-cell epitopes exist in 41 kD flagellin of B. burgdorferi sensu lato strains; these changes should be considered in serodiagnosis of Lyme disease.

  18. Five-month comparative efficacy evaluation of three ectoparasiticides against adult cat fleas (Ctenocephalides felis), flea egg hatch and emergence, and adult brown dog ticks (Rhipicephalus sanguineus sensu lato) on dogs housed outdoors.

    Science.gov (United States)

    Varloud, Marie; Hodgkins, Elizabeth

    2015-03-01

    This study was designed to compare the efficacy of three topical combinations on dogs in outdoor conditions against adult cat fleas (Ctenocephalides felis), flea egg hatch and emergence, and against adult brown dog ticks (Rhipicephalus sanguineus sensu lato). Treatment was performed on day 0 with a placebo; dinotefuran, pyriproxifen and permethrin (DPP); fipronil and (S)-methoprene (FM) or imidacloprid and permethrin (IP). Dogs (n = 32), housed outdoors for 7 months, were treated monthly for four consecutive months (on days 0, 30, 60 and 90) and infested with ~100 unfed adult fleas on days 14, 55, 74, 115 and 150 and with ~50 unfed adult ticks on days 28, 44, 88 and 104. Adult fleas were counted and removed 24 h after infestation. Immediately after flea removal, dogs were reinfested with ~100 new adult fleas 72 h prior to egg collection for up to 48 h. Flea eggs were incubated for 32 days, and newly emerged adults were counted. Ticks were counted and removed 48 h after each infestation. FM had >90 % efficacy against fleas at each time point and variable efficacy against ticks (38.0-99.6 %). Efficacy of IP was 60 days after the last treatment. Despite challenging weather conditions, DPP was highly effective, providing >90 % efficacy against adult ticks as well as adult and immature fleas at every time point of the study.

  19. Prevalence of Borrelia burgdorferi infection in a series of 98 primary cutaneous lymphomas.

    Science.gov (United States)

    Ponzoni, Maurilio; Ferreri, Andrés J M; Mappa, Silvia; Pasini, Elisa; Govi, Silvia; Facchetti, Fabio; Fanoni, Daniele; Tucci, Alessandra; Vino, Arianna; Doglioni, Claudio; Berti, Emilio; Dolcetti, Riccardo

    2011-01-01

    Borrelia burgdorferi has been variably associated with different forms of primary cutaneous lymphoma. Differences in prevalence rates among reported studies could be a result of geographic variability or heterogeneity in the molecular approaches that have been employed. In the present study, we investigated the prevalence of Borrelia burgdorferi sensu lato DNA in diagnostic tissue samples from fresh cutaneous biopsies of 98 primary cutaneous lymphomas and 19 normal skin controls. Three different polymerase chain reaction (PCR) protocols targeting the hbb, flagellin, and Osp-A genes were used. Direct sequencing of both sense and antisense strands of purified PCR products confirmed the specificity of the amplified fragments. Sequence specificity was assessed using the Basic Local Alignment Search Tool, and MultAlin software was used to investigate the heterogeneity of target gene sequences across the different samples. Borrelia DNA was not detected in 19 controls, 23 cases of follicular lymphoma, 31 cases of extranodal marginal zone lymphoma, or 30 cases of mycosis fungoides. A single case of 14 diffuse large B-cell lymphoma cases was positive for B. burgdorferi. This study does not support a pathogenic role of B. burgdorferi in primary cutaneous B- and T-cell lymphomas from areas nonendemic for this microorganism and the consequent rationale for the adoption of antibiotic therapy in these patients.

  20. Residência multiprofissional em saúde e pós-graduação lato sensu no Brasil: apontamentos históricos Multiprofessional residency in healthcare and postgraduate education in Brazil: historical remarks

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    Soraya Diniz Rosa

    2009-11-01

    Full Text Available Trata-se da discussão da educação superior brasileira, tomando-se como parâmetro a especialização no conjunto da pós-graduação, a partir da análise do Programa da Residência Multiprofissional em Saúde, reconhecido como pós-graduação lato sensu. Utilizou-se a pesquisa histórica e documental como instrumento para análise dos problemas enfrentados no modelo de formação proposto no referido programa e apresentado pelos ministérios da Educação e da Saúde como base para a consolidação da política nacional de educação em saúde. Os resultados alcançados nos permitem afirmar a importância da pós-graduação lato sensu no âmbito da educação em saúde. Contudo, sua oferta descontínua e, na maioria dos casos, não acadêmica, tem caracterizado um modelo limitado pela lógica mercantil, evidenciando a falta de políticas públicas que assegurem a devida qualificação profissional. A Residência Multiprofissional em Saúde pretende ser uma nova estratégia para políticas de educação permanente que, enfocando categorias profissionais não médicas da área, favoreça a produção das condições necessárias para mudanças no modelo médico-assistencial restritivo, ainda hegemônico, de atenção em saúde. Resta saber se essa interferência no modelo educacional vai contribuir para uma melhor ação profissional ou se restringe à estruturação da rede de serviços públicos, através da oferta de um campo de trabalho precarizado.This is a discussion about Brazilian higher education, using as a parameter specialization courses in the postgraduate program based on the analysis of the Programa da Residência Multiprofissional em Saúde [Multidisciplinary Residency Program in Health-care], known as a postgraduate program. We used historical and documental research as a tool for analysis of the problems faced in the training model proposed in the Program and presented by the ministries of Education and Health as the basis

  1. Genetic characterization of human-derived hydatid cysts of Echinococcus granulosus sensu lato in Heilongjiang Province and the first report of G7 genotype of E. canadensis in humans in China.

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    Tiemin Zhang

    Full Text Available Cystic echinococcosis (CE caused by the larval stage of Echinococcus granulosus sensu lato (s.l. is one of the most important zoonotic parasitic diseases worldwide and 10 genotypes (G1-G10 have been reported. In China, almost all the epidemiological and genotyping studies of E. granulosus s.l. are from the west and northwest pasturing areas. However, in Heilongjiang Province of northeastern China, no molecular information is available on E. granulosus s.l. To understand and to speculate on possible transmission patterns of E. granulosus s.l., we molecularly identified and genotyped 10 hydatid cysts from hepatic CE patients in Heilongjiang Province based on mitochondrial cytochrome c oxidase subunit I (cox1, cytochrome b (cytb and NADH dehydrogenase subunit 1 (nad1 genes. Two genotypes were identified, G1 genotype (n = 6 and G7 genotype (n = 4. All the six G1 genotype isolates were identical to each other at the cox1 locus; three and two different sequences were obtained at the cytb and nad1 loci, respectively, with two cytb gene sequences not being described previously. G7 genotype isolates were identical to each other at the cox1, cytb and nad1 loci; however, the cytb gene sequence was not described previously. This is the first report of G7 genotype in humans in China. Three new cytb gene sequences from G1 and G7 genotypes might reflect endemic genetic characterizations. Pigs might be the main intermediate hosts of G7 genotype in our investigated area by homology analysis. The results will aid in making more effective control strategies for the prevention of transmission of E. granulosus s.l.

  2. Genetic characterization of human-derived hydatid cysts of Echinococcus granulosus sensu lato in Heilongjiang Province and the first report of G7 genotype of E. canadensis in humans in China.

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    Zhang, Tiemin; Yang, Dong; Zeng, Zhaolin; Zhao, Wei; Liu, Aiqin; Piao, Daxun; Jiang, Tao; Cao, Jianping; Shen, Yujuan; Liu, Hua; Zhang, Weizhe

    2014-01-01

    Cystic echinococcosis (CE) caused by the larval stage of Echinococcus granulosus sensu lato (s.l.) is one of the most important zoonotic parasitic diseases worldwide and 10 genotypes (G1-G10) have been reported. In China, almost all the epidemiological and genotyping studies of E. granulosus s.l. are from the west and northwest pasturing areas. However, in Heilongjiang Province of northeastern China, no molecular information is available on E. granulosus s.l. To understand and to speculate on possible transmission patterns of E. granulosus s.l., we molecularly identified and genotyped 10 hydatid cysts from hepatic CE patients in Heilongjiang Province based on mitochondrial cytochrome c oxidase subunit I (cox1), cytochrome b (cytb) and NADH dehydrogenase subunit 1 (nad1) genes. Two genotypes were identified, G1 genotype (n = 6) and G7 genotype (n = 4). All the six G1 genotype isolates were identical to each other at the cox1 locus; three and two different sequences were obtained at the cytb and nad1 loci, respectively, with two cytb gene sequences not being described previously. G7 genotype isolates were identical to each other at the cox1, cytb and nad1 loci; however, the cytb gene sequence was not described previously. This is the first report of G7 genotype in humans in China. Three new cytb gene sequences from G1 and G7 genotypes might reflect endemic genetic characterizations. Pigs might be the main intermediate hosts of G7 genotype in our investigated area by homology analysis. The results will aid in making more effective control strategies for the prevention of transmission of E. granulosus s.l.

  3. 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.

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

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    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%).

  5. Prevalence of Borrelia burgdorferi in Ixodes ricinus ticks collected from moose (Alces alces) and roe deer (Capreolus capreolus) in southern Norway.

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    Kjelland, Vivian; Ytrehus, Bjørnar; Stuen, Snorre; Skarpaas, Tone; Slettan, Audun

    2011-06-01

    As part of a larger survey, ears from 18 roe deer (Capreolus capreolus) and 52 moose (Alces alces) shot in the 2 southernmost counties in Norway were collected and examined for Ixodes ricinus ticks. Seventy-two adult ticks, 595 nymphs, and 267 larvae from the roe deer, and 182 adult ticks, 433 nymphs, and 70 larvae from the moose were investigated for infection with Borrelia burgdorferi sensu lato (s.l.). The results showed the presence of B. burgdorferi s.l. DNA in 2.9% of the nymphs collected from roe deer and in 4.4% of the nymphs and 6.0% of the adults collected from moose. The spirochetes were not detected in adult ticks from roe deer, or in larvae feeding on roe deer or moose. In comparison, the mean infection prevalences in questing I. ricinus collected from the same geographical area were 0.5% infection in larvae, 24.5% in nymphs, and 26.9% in adults. The most prevalent B. burgdorferi genospecies identified in ticks collected from roe deer was B. afzelii (76.5%), followed by B. garinii (17.6%), and B. burgdorferi sensu stricto (5.9%). Only B. afzelii (76.7%) and B. garinii (23.3%) were detected in ticks collected from moose. The present study indicates a lower prevalence of B. burgdorferi infection in I. ricinus ticks feeding on roe deer and moose compared to questing ticks. This is the first study to report B. burgdorferi s.l. prevalence in ticks removed from cervids in Norway.

  6. 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.

  7. Borrelia burgdorferi Manipulates Innate and Adaptive Immunity to Establish Persistence in Rodent Reservoir Hosts

    Science.gov (United States)

    Tracy, Karen E.; Baumgarth, Nicole

    2017-01-01

    Borrelia burgdorferi sensu lato species complex is capable of establishing persistent infections in a wide variety of species, particularly rodents. Infection is asymptomatic or mild in most reservoir host species, indicating successful co-evolution of the pathogen with its natural hosts. However, infected humans and other incidental hosts can develop Lyme disease, a serious inflammatory syndrome characterized by tissue inflammation of joints, heart, muscles, skin, and CNS. Although B. burgdorferi infection induces both innate and adaptive immune responses, they are ultimately ineffective in clearing the infection from reservoir hosts, leading to bacterial persistence. Here, we review some mechanisms by which B. burgdorferi evades the immune system of the rodent host, focusing in particular on the effects of innate immune mechanisms and recent findings suggesting that T-dependent B cell responses are subverted during infection. A better understanding of the mechanisms causing persistence in rodents may help to increase our understanding of the pathogenesis of Lyme disease and ultimately aid in the development of therapies that support effective clearance of the bacterial infection by the host’s immune system. PMID:28265270

  8. 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.

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

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    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.

  10. A catalog of the types of Stratiomyidae, Tabanidae, Asilidae sensu lato, and Syrphidae (Insecta, Diptera deposited in the Museo Argentino de Ciencias Naturales, Buenos Aires (MACN: with an appendix on types of Tabanidae deposited in the Instituto "Dr. Carlos G. Malbrán", Buenos Aires (ANLIS

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    Axel O Bachmann

    2012-06-01

    Full Text Available The type specimens of all accepted categories of three names of Stratiomyidae, fifty-nine of Tabanidae, twenty-four of Asilidae sensu lato (includes Leptogastridae, and twenty of Syrphidae are recorded. In an appendix twelve paratypes of six species of Tabanidae are recorded, housed in the Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán" (ANLIS, Departamento Vectores, Buenos Aires.Se registran los ejemplares típo de todas las categorías aceptadas de tres nombres de Stratiomyidae, cincuenta y nueve de Tabanidae, veinticuatro de Asilidae s. l. (incluye Leptogastridae y veinte de Syrphidae. En un appendix se registran doce paratipos, de seis especies de Tabanidae, depositados en la Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán" (ANLIS, Departamento Vectores, Buenos Aires.

  11. 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

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

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    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.

  13. 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.

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

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    Brandon L Garcia

    2016-01-01

    Full Text Available 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.

  15. 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.

  16. Review of the taxonomy of the genus Arthrobacter, emendation of the genus Arthrobacter sensu lato, proposal to reclassify selected species of the genus Arthrobacter in the novel genera Glutamicibacter gen. nov., Paeniglutamicibacter gen. nov., Pseudoglutamicibacter gen. nov., Paenarthrobacter gen. nov. and Pseudarthrobacter gen. nov., and emended description of Arthrobacter roseus.

    Science.gov (United States)

    Busse, Hans-Jürgen

    2016-01-01

    In this paper, the taxonomy of the genus Arthrobacter is discussed, from its first description in 1947 to the present state. Emphasis is given to intrageneric phylogeny and chemotaxonomic characteristics, concentrating on quinone systems, peptidoglycan compositions and polar lipid profiles. Internal groups within the genus Arthrobacter indicated from homogeneous chemotaxonomic traits and corresponding to phylogenetic grouping and/or high 16S rRNA gene sequence similarities are highlighted. Furthermore, polar lipid profiles and quinone systems of selected species are shown, filling some gaps concerning these chemotaxonomic traits. Based on phylogenetic groupings, 16S rRNA gene sequence similarities and homogeneity in peptidoglycan types, quinone systems and polar lipid profiles, a description of the genus Arthrobacter sensu lato and an emended description of Arthrobacter roseus are provided. Furthermore, reclassifications of selected species of the genus Arthrobacter into novel genera are proposed, namely Glutamicibacter gen. nov. (nine species), Paeniglutamicibacter gen. nov. (six species), Pseudoglutamicibacter gen. nov. (two species), Paenarthrobacter gen. nov. (six species) and Pseudarthrobacter gen. nov. (ten species).

  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. 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. A comparison of Mg/Ca ratios in Globigerinoides ruber (white): sensu stricto versus a mixture of genotypes

    Digital Repository Service at National Institute of Oceanography (India)

    Naik, S.S.

    Mg/Ca ratios were measured in core AAS9/21 from the eastern Arabian Sea (EAS), on two sets of planktonic foraminifera, one with a mixture of Globigerinoides ruber genotypes (sensu stricto and sensu lato) and the second which contained only G. ruber...

  20. Candida parapsilosis sensu stricto and the closely related species Candida orthopsilosis and Candida metapsilosis in vulvovaginal candidiasis.

    Science.gov (United States)

    Zhu, Yuxia; Shan, Yingying; Fan, Shangrong; Li, Jianling; Liu, Xiaoping

    2015-02-01

    This study aimed to determine the clinical characteristics and in vitro susceptibilities of Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis isolates from patients with vulvovaginal candidiasis (VVC). We analysed 63 vaginal C. parapsilosis specimens. After the molecular analyses, the isolates were characterised as C. parapsilosis sensu stricto (77.8%), C. orthopsilosis (7.9%) and C. metapsilosis (14.3%). The signs and symptoms of VVC caused by C. parapsilosis sensu lato, including itching, erythema and abnormal discharge, were milder than those caused by C. albicans. None of the C. parapsilosis sensu lato isolates were resistant to fluconazole, miconazole or itraconazole. The resistance rates of C. albicans to fluconazole, itraconazole, miconazole and clotrimazole were 2.3, 1.5, 3.1 and 0.8%, respectively. Both C. parapsilosis sensu lato and C. albicans were susceptible to nystatin. The mycological eradication rate at follow-up days 7-14 and 30-35 were 77.8% (49/63) and 76.2% (48/63), respectively, when treated with various antifungal agents and regimens. We conclude that C. parapsilosis sensu stricto and the closely related species C. orthopsilosis and C. metapsilosis were present in the vaginal samples of VVC patients. The symptoms and signs of VVC caused by C. parapsilosis are milder than those caused by C. albicans. The antifungal susceptibility and therapeutic efficacy in patients colonised by C. parapsilosis sensu lato were similar to those observed in C. albicans-colonised patients.

  1. Phylogeny of Elymus sensu lato in Triticeae (Poaceae) based on plastid trnL-F sequence data%基于叶绿体trnL-F序列对广义披碱草属物种的系统进化研究

    Institute of Scientific and Technical Information of China (English)

    张春; 凡星; 沙莉娜; 康厚扬; 张海琴; 周永红

    2011-01-01

    The genus Elymus sensu lato includes Roegneria, Elymus, Hystrix, Sitanion, and Kengyilia. The plastid trnL-F sequence data were analyzed for their phylogenetic relationships, maternal donor, and for the origin and differentiation of St genomes by using Maximum Parsimony (MP) and Median Joining algorithm (MJ) methods. 1) The maternal donor of Elymus sensu lato was not always Pseudoroegneria. The P genome served as the maternal donor to Kengyilia melanthera and K. thoroldiana, while the Ns genome was the maternal donor for Hystrix duthiei and H. duthiei ssp. longearistata; 2) The MP tree revealed the close relationships between P, W and St genomes, while the H and Ns genomes were distinctly related to them; 3) K. melanthera and K. thoroldiana were clustered with diploid Agropyron species, which indicated the close relationships among them. H. patula was clustered in the St clade, while H. duthiei and H. duthiei ssp. longearistata were grouped in the Ns clade. The genomic constitution of H. patula was different from H. duthiei and H. duthiei ssp. longearistata; 4) The five diploid species with St genome did not form the monophyletic clades, suggesting the differentiation of the diploid species. The olyploid species with St genome were not clustered together, suggesting the differentiation of St genomes in polyploids; 5) The species distributed in Eurasia might originate from the diploid species of Pseudoroegneria from Asia, while the North American species might originate from North America Pseudoroegneria species. Some species might have a multiple origin and experienced recurrent hybridization between species with different genomes.%广义披碱草属包括了披碱草属、鹅观草属、猬草属、裂颖草属和仲彬草属.为深入研究这5个小麦族多年生属的系统地位、母本来源及St染色体组的来源和分化等问题,本研究根据叶绿体DNA trnL-F序列构建了系统树和网状结构图.结果表明,1)广义披碱草属物种的母

  2. Observaciones cariológicas en Vicia amphicarpa sensu lato

    Directory of Open Access Journals (Sweden)

    Tardío Pato, F. Javier

    1998-12-01

    Full Text Available Cytological studies were carried out on twenty four amphicarpic wild populations of Vicia sativa group from different sites (19 from Spain, 4 from Syria and 1 from Turkey. Morphologically, 19 accessions can be attributed to taxon Dorthes and 5 to V. angustifolia subsp. pseudoangustifolia (Rouy Tardío, comb. nov. In our studies, these two taxa show different chromosome numbers, V. amphicarpa Dorthes, has 14 chromosomes, while V. angustifolia subsp. pseudoangustifolia has only 12. Furthermore, the form of the chromosomes in the karyotypes of both taxa is quite different. The chromosome number and the morphology of the karyotype found in the first taxon agree with most of previous reports on V. amphicarpa, with a very characteristic short metacentric and six acrocentric chromosomes, one of them satellited. However, the second taxon has a very different karyotype with three large chromosomes, one of them submetacentric and another satellited, plus another three chromosomes which are significantly smaller. Therefore, we can conclude that within the genus Vicia, at least two amphicarpic taxa exist, clearly differentiated, both in morphological and karyotypical characteristics.Se estudian cariológicamente 24 poblaciones anficárpicas silvestres pertenecientes al grupo Vicia sativa, de distinta procedencia (19 españolas, cuatro de Siria y una de Turquía. Se encuentran diferencias en los números cromosomáticos y en los cariótipos. Diecinueve poblaciones presentan 14 cromosomas y cariotipo como el observado por otros autores, constituido por un pequeño cromosoma metacéntrico y seis acrocéntricos, uno de ellos con satélite. Éstas corresponden a V. amphicarpa Dorthes. Las otras cinco, con 2n = 12 y cariotipo con tres cromosomas grandes, uno de ellos submetacéntrico, otro con satélite, y otros tres mucho más pequeños, corresponden a V. angustifolia subsp. pseudoangustifolia (Rouy Tardío. Esta anterior se propone como combinación nueva. Por lo tanto, dentro de Vicia existen, al menos, dos táxones anficárpicos claramente diferenciados, tanto en su morfología externa como en el cariotipo.

  3. Do ticks and Borrelia burgdorferi s.l. constitute a burden to birds?

    Science.gov (United States)

    Norte, A C; Lobato, D N C; Braga, E M; Antonini, Y; Lacorte, G; Gonçalves, M; Lopes de Carvalho, I; Gern, L; Núncio, M S; Ramos, J A

    2013-05-01

    Ticks consume resources from their hosts shaping their life-history traits and are vectors of many zoonotic pathogens. Several studies have focused on the health effects of blood-sucking ectoparasites on avian hosts, but there is limited information on the effects of ticks on adult and sub-adult birds, which may actively avoid ticks and are likely to present low infestation intensities. We evaluated the effects of the presence of feeding ticks and intensity of infestation on health variables of avian hosts. We also evaluated whether these variables were affected by tick infection by Borrelia burgdorferi sensu lato (s.l.) and by the presence of Borrelia infection on the birds' skin. Presence of parasite association among ticks, haemosporidea and Borrelia within the bird-host was also tested. We found that infestation by ticks significantly increased heterophyl/lymphocyte ratio in Turdus merula suggesting increased stress. This was especially evident at high infestation intensities when a significant decrease in body mass and body condition (body mass corrected for size) was also observed. Erithacus rubecula infested with more than 10 larvae tended to have lower haematocrit and blood haemoglobin. Plasma globulin concentration in T. merula tended to be affected by the presence of attached ticks and their infection with Borrelia, but this depended on the age of the bird. No association was detected among ticks, haemosporidea and Borrelia infection. We showed that ticks have detrimental effects on their avian hosts even under natural infestation conditions and that confirmed Borrelia reservoir hosts may also present symptoms of infection, though these may be subtle.

  4. Asymptomatic Borrelia burgdorferi infection.

    Science.gov (United States)

    Wormser, G P; Nadelman, R B; Nowakowski, J; Schwartz, I

    2001-10-01

    Little is known about the natural history of asymptomatic Borrelia burgdorferi infection. Our analysis of the asymptomatic infections diagnosed serologically in a recent OspA vaccine trial conducted in the United States (N Engl J Med 1998;339: 209-215), suggests that the natural history of this event is more benign than that reported for untreated patients with erythema migrans (Ann Intern Med 1987;107: 725-731). We hypothesize that this is due either to incorrect diagnosis since the specificity of the serologic criteria used to diagnose asymptomatic infection in the vaccine study is unknown, or to infection with non-pathogenic strains of B. burgdorferi. Increasing evidence indicates that the invasive potential of strains of B. burgdorferi varies according to the specific subtype. Theoretically, a serologic testing method could be devised which would distinguish infection with invasive versus non-invasive strains of B. burgdorferi, and allow testing of the second hypothesis.

  5. 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.

  6. Identification of a New Borrelia Species among Small Mammals in Areas of Northern Spain Where Lyme Disease Is Endemic

    Science.gov (United States)

    Gil, Horacio; Barral, Marta; Escudero, Raquel; García-Pérez, Ana L.; Anda, Pedro

    2005-01-01

    The role of small mammals as reservoir hosts for Borrelia burgdorferi was investigated in several areas where Lyme disease is endemic in northern Spain. A low rate of infestation by Ixodes ricinus nymphs was found in the small mammal populations studied that correlated with the near-absence of B. burgdorferi sensu lato in 184 animals tested and with the lack of transmission of B. burgdorferi sensu lato to I. ricinus larvae that fed on them. In contrast, questing ticks collected at the same time and in the same areas were found to carry a highly variable B. burgdorferi sensu lato repertoire (B. burgdorferi sensu stricto, Borrelia garinii, Borrelia valaisiana, and Borrelia afzelii). Interestingly, the only isolate obtained from small mammals (R57, isolated from a bank vole) grouped by phylogenetic analyses with other Borrelia species but in a separate clade from the Lyme disease and relapsing fever organisms, suggesting that it is a new species. This new agent was widely distributed among small mammals, with infection rates of 8.5 to 12% by PCR. Moreover, a high seroprevalence to B. burgdorferi sensu lato was found in the animal sera, suggesting cross-reactivity between B. burgdorferi sensu lato and R57. Although small mammals do not seem to play an important role as reservoirs for B. burgdorferi sensu lato in the study area, they seem to be implicated in the maintenance of spirochetes similar to R57. PMID:15746336

  7. 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.

  8. Classification of Italian isolates of Borrelia burgdorferi into three genomic groups.

    Science.gov (United States)

    Cinco, M; De Giovannini, R; Fattorini, P; Florian, F; Graziosi, G

    1993-10-01

    In this study we investigated the genotypic characteristics of some locally isolated strains of B. burgdorferi by three different methodologies: restriction endonuclease analysis (REA), Southern blot hybridization with whole DNAs from Borrelia strains and Southern blot hybridization with rRNA 16 + 23S genes derived from E. coli. REA fingerprintings were evaluated by cluster analysis, according to the principles of numerical taxonomy. The genomas of the locally isolated strains were compared with borreliae originating from different countries of Europe, including Sweden and with the American reference strain B31. Among the European strains, some already described by Baranton (Baranton et al., 1992) as representatives of different genomic groups Borrelia sensu stricto and Borrelia garinii were used. By the different techniques the isolates were included in three genomic groups which could correspond to the three genospecies identified by Baranton, namely B. burgdorferi sensu stricto, B. garinii and B. group VS461: in fact two strains were included in a homogeneous group, probably corresponding to the VS461 genomic group, together with other European borreliae; one isolate was included in a group consisting of B31 and some other European strains already described as belonging to Borrelia burgdorferi in sensu stricto. Finally two isolates were ascribed to a third genomic group probably corresponding to the genospecies indicated as Borrelia garinii. These findings indicate that a small number of Borrelia strains isolated from a very restricted area can be genetically heterogeneous.

  9. La familia Apocynaceae sensu lato en México: diversidad y distribución The family Apocynaceae sensu lato in Mexico: diversity and distribution

    Directory of Open Access Journals (Sweden)

    Verónica Juárez-Jaimes

    2007-12-01

    Full Text Available La familia Apocynaceae s.l. se ubica entre las 15 familias más diversas de México, con 385 especies repartidas en 50 géneros y 3 subfamilias, de las cuales Asclepiadoideae es la de mayor diversidad en géneros, especies y endemismos. Los géneros con más especies nativas (y endémicas son Asclepias y Matelea. Los estados más ricos en diversidad de taxones son los ubicados en ambas vertientes; destacan Chiapas, Guerrero, Oaxaca y Veracruz. Por su número de especies, sobresalen el bosque tropical caducifolio y los matorrales xerófilos. Del total de especies, 176 son endémicas de México. Se presenta la lista de especies registradas, señalándose su distribución por estados.Apocynaceae s.l. is among the 15 most diverse families in Mexico, with a total of 385 species, distributed in 50 genera and 3 subfamilies of which Asclepiadoideae is the most diverse in numbers of genera, species and endemics. The genera with the most native species and endemics are Asclepias and Matelea. The richest states in terms of taxon diversity are those at the Pacific and Atlantic slopes, Chiapas, Oaxaca, Guerrero and Veracruz being the most prominent. Tropical deciduous forest and arid tropical scrub are the most important vegetation types by their total number of species, 176 of which are endemic to Mexico. A list of species recorded in México is provided, indicating their state distribution.

  10. 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 (

  11. Microdiversity of Echinococcus granulosus sensu stricto in Australia.

    Science.gov (United States)

    Alvarez Rojas, C A; Ebi, D; Gauci, C G; Scheerlinck, J P; Wassermann, M; Jenkins, D J; Lightowlers, M W; Romig, T

    2016-07-01

    Echinococcus granulosus (sensu lato) is now recognized as an assemblage of cryptic species, which differ considerably in morphology, development, host specificity (including infectivity/pathogenicity for humans) and other aspects. One of these species, E. granulosus sensu stricto (s.s.), is now clearly identified as the principal agent causing cystic echinococcosis in humans. Previous studies of a small section of the cox1 and nadh1 genes identified two variants of E. granulosus s.s. to be present in Australia; however, no further work has been carried out to characterize the microdiversity of the parasite in its territory. We have analysed the sequence of the full length of the cox1 gene (1609 bp) from 37 isolates of E. granulosus from different hosts and geographic regions of Australia. The analysis shows that seven haplotypes of E. granulosus s.s. not previously described were found, together with five haplotypes known to be present in other parts of the world, including the haplotype EG01 which is widespread and present in all endemic regions. These data extend knowledge related to the geographical spread and host range of E. granulosus s.s. in a country such as Australia in which the parasite established around 200 years ago.

  12. A new approach to identify species in Xiphinema americanum sensu lato.

    Science.gov (United States)

    Prior, T; Hockland, S; Decraemer, W

    2010-01-01

    Four putative species belonging to the X. americanum group are known to transmit American nepoviruses and these nematodes and viruses are listed in European quarantine legislation. Identification of species in this group is therefore of particular importance for phytosanitary purposes, but is problematic because of the similar morphology of the putative species. As part of the Synthesys project BE-TAF 1769, eight collaborative institutions contributed material to Fera for study. Video technology allowed the best records of type material and in conjunction with photographic images created a 'virtual' collection of images that relies less on the deteriorating quality of museum specimens. Revised definitions of lip region and tail shape are considered to be the most stable characters for differentiation. Position of the amphidial aperture, development of the odontostyle collar and some established morphometric characters are considered of limited use although they retain value for latter stages of identification.

  13. Morphometric Evidence for Three Juvenile Stages in Some Species of Xiphinema americanum sensu lato.

    Science.gov (United States)

    Halbrendt, J M; Brown, D J

    1992-06-01

    One to two hundred nematodes from each of seven Xiphinema americanum-group populations were measured to determine the range of stylet and body lengths for juveniles and adults. First-stage juveniles were identified by the position of the replacement odontostyle (i.e., the tip of the replacement odontostyle overlapped the base of the odontophore). Nematodes were identified as second stage if the functional odontostyle was the same length as the replacement odontostyle of the first stage. Subsequent stages were similarly identified by establishing the range of corresponding replacement and functional odontostyle lengths. In all populations examined, this procedure created natural divisions that clearly grouped nematodes by stylet and body length. Presumably these groups identified all juvenile and adult stages. Populations of X. americanum, X. rivesi, and X. californicum from the United States had three juvenile stages, but a population of X. pachtaicum from Bulgaria had four juvenile stages.

  14. Downy mildew on coleus (Plectranthus scutellarioides) caused by Peronospora belbahrii sensu lato in Tennessee

    Science.gov (United States)

    Coleus (Plectranthus scutellarioides [syn. = Solenostemon scutellarioides]) is a popular ornamental plant in the mint family (Lamiaceae), prized for its colorful and showy foliage. In August 2015, disease symptoms typical of downy mildew were observed at two sites in Nashville, Tennessee: (i) at the...

  15. Use of DNA barcode sensu lato to identify traditional Tibetan medicinal plant Gentianopsis paludosa (Gentianaceae)

    Institute of Scientific and Technical Information of China (English)

    Chun-Ying XUE; De-Zhu LI

    2011-01-01

    Gentianopsis paludosa (Hook. f.) Ma (Gentianaceae) is an important species in Tibetan folk medicine commonly used to clear away the "heat evils" and toxic materials. A survey of market samples revealed that nine adulterant species, Gentianopsis barbata, G. contorta, G. grandis, Halenia elliptica, Lomatogonium macranthum, L.rotatum, Swertia angustifolia, S. bifolia and S. erythrosticta, are often marketed as G. paludosa. Methods to distinguish G. paludosa from its adulterants are limited by current morphological and chemical methods. DNA sequence analysis of the nuclear ribosomal DNA internal transcribed spacer region (ITS) was used in the differentiation of G.paludosa from the adulterant species. The data showed that the internal transcribed spacer regions differ significantly between G. paludosa and all nine adulterant species, so that they could be easily distinguished at the DNA level.

  16. Flora of the Xingo region, Alagoas-Sergipe: Portulacaceae sensu lato

    Directory of Open Access Journals (Sweden)

    Hermes de Oliveira Machado-Filho

    2012-11-01

    Full Text Available Portulacaceae Juss. Includes herbaceous to subshrub plants, presenting fleshy leaves, alternate, sub-opposite to opposite, bisexual flowers, actinomorphic, with or without bracts, and fruit usually in capsule with longitudinal or transverse dehiscence. This paper deals with the floristic-taxonomic study of Portulacaceae in the Xingo region, in the states of Alagoas and Sergipe (Northeastern Brazil. We recorded 2 genera and 6 species: Portulaca L., with 4 species (P. elatior Mart. ex Rohrb., P. halimoides L., P. mucronata Link, and P. oleracea L. and Talinum Adans., with 2 species (T. paniculatum (Jacq. Gaertn. and T. triangulare (Jacq. Willd.. We present descriptions, illustrations, and keys for separation of genera and species, besides distribution and habitat data on the species found in the area.

  17. Molecular phylogenies reveal host-specific divergence of Ophiocordyceps unilateralis sensu lato following its host ants.

    Science.gov (United States)

    Kobmoo, N; Mongkolsamrit, S; Tasanathai, K; Thanakitpipattana, D; Luangsa-Ard, J J

    2012-06-01

    Ophiocordyceps unilateralis (Hypocreales, Ascomycetes) is an entomopathogenic fungus specific to formicine ants (Formicinae, Hymenoptera). Previous works have shown that the carpenter ant Camponotus leonardi acts as the principal host with occasional infections of ants from the genus Polyrhachis (sister genus of Camponotus). Observations were made on the permanent plots of Mo Singto, Khao Yai National Park of Thailand according to which O. unilateralis was found to occur predominantly on three host species: C. leonardi, C. saundersi and P. furcata. Molecular phylogenies of the elongation factor 1-α and β-Tubulin genes indicate a separation of O. unilateralis samples into three clades, reflecting specificity to each of the three different ant species. Samples collected from P. furcata and from C. leonardi were found to form sister groups with samples from C. saundersi forming an outgroup to the latter. Additional samples collected from unidentified ant species of Camponotus and Polyrhachis were positioned as outgroups to those samples on identified species. These results demonstrate that O. unilateralis is clearly not a single phylogenetic species and comprises at least three species that are specific to different host ant species. These cryptic species may arise through recent events of speciation driven by their specificity to host ant species.

  18. 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

  19. Molecular epidemiology and antifungal susceptibility of Candida parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis in Taiwan.

    Science.gov (United States)

    Chen, Yee-Chun; Lin, Yu-Hsin; Chen, Kuo-Wei; Lii, Joanne; Teng, Hwa-Jen; Li, Shu-Ying

    2010-11-01

    Candida parapsilosis was recently reclassified into 3 closely related species, C. parapsilosis sensu stricto, Candida orthopsilosis, and Candida metapsilosis. Variation in susceptibility characteristics and prevalence of the 3 genomic species could have therapeutic and epidemiologic implications. The aim of this study is to characterize the genetic and antifungal susceptibility profiles of 97 C. parapsilosis isolates from 71 patients. Among the 71 nonduplicate isolates, 85.9% (61/71) were identified as C. parapsilosis sensu stricto, 5.6% (4/71) as C. metapsilosis, and 8.5% (6/71) as C. orthopsilosis species based on sequences of the internal transcribed spacer (ITS) region. The delineation of these 3 species is concordant with that achieved by pulsed-field gel electrophoresis of BssHII restriction fragments at 75% similarity. Antifungal susceptibility tests showed that most isolates were susceptible to flucytosine, azoles, amphotericin B, and echinocandins, whereas 3 C. metapsilosis isolates from 1 patient showed resistance and susceptible-dose dependence to fluconazole. The C. metapsilosis isolates exhibited significantly higher MIC values to both fluconazole and voriconazole than those of C. parapsilosis sensu stricto and C. orthopsilosis. On the other hand, the C. metapsilosis isolates showed significantly lower MIC values on 24 h to caspofungin than those of C. parapsilosis sensu stricto and C. orthopsilosis. For micafungin, the isolates of C. parapsilosis sensu stricto had significantly higher MIC values on 24 h than those of C. orthopsilosis and C. metapsilosis. Compared to Candida albicans, mutations from proline to alanine were identified on the hot spot 1 of Fks1 in all these C. parapsilosis sensu lato isolates regardless of their MIC levels. Some of the C. orthopsilosis and C. metapsilosis isolates expressed the isoleucine to valine substitution on the hot spot 2 region. However, the amino acid variations in these isolates did not correlate to their MIC

  20. Echinococcus granulosus sensu stricto (s.s.) from the critically endangered antelope Addax nasomaculatus in Tunisia.

    Science.gov (United States)

    Boufana, Belgees; Saïd, Yousra; Dhibi, Mokhtar; Craig, Philip S; Lahmar, Samia

    2015-12-01

    Echinococcus granulosus sensu lato (s.l.) is a zoonotic disease highly endemic in Tunisia. Canids including stray and semi-stray dogs, jackals and foxes are known as definitive hosts and a wide range of ungulates have been shown to harbour the metacestode hydatid stage and may serve as intermediate hosts. Fertile hydatid cysts of Echinococcus equinus and E. granulosus sensu stricto (s.s.) were recently molecularly identified for the first time from Tunisian donkeys. E. granulosus (s.s.) was also identified from wild boars in Tunisia. Here we report the confirmation of hydatid cysts caused by E. granulosus (s.s.) in the critically endangered antelope, Addax nasomaculatus in Tunisia. DNA-based molecular analysis revealed that A.nasomaculatus was infected with E. granulosus (s.s.) which had a 100% identity with the main globally distributed E. granulosus (s.s.) (EgTu01) haplotype. Cysts of Taenia hydatigena (n=33) were also observed on the liver and in the body cavity. Due to their endangered status and their relatively small numbers, it is unlikely that hydatid infection of A. nasomaculatus will form a major contribution to the epidemiology and transmission of E. granulosus in Tunisia, but infection may result in pathology, morbidity and early mortality, and may still play a role in the perpetuation of the parasite in wildlife cycles.

  1. 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.

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

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    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.

  3. Estrutura da comunidade de Chlorococcales sensu lato (Chlorophyceae em diferentes hábitats aquáticos e hidroperíodos Structure of the Chlorococcales sensu lato (Chlorophyceae community in different aquatic habitats and hydroperiods

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    Lezilda Carvalho Torgan

    2011-03-01

    Full Text Available Este estudo visa avaliar a riqueza, a densidade, a diversidade específica e a distribuição da comunidade de Chlorococcales s.l. em zonas pelágicas e litorâneas de diferentes hábitats aquáticos (lagoas isoladas, lagoas interligadas, banhados, açudes e arroios em dois hidroperíodos (águas altas e baixas em região subtropical no sul do Brasil. As amostragens de fitoplâncton foram realizadas em duas áreas, situadas entre as coordenadas 30(040´ - 30(010´ S e 50(030´ - 51(031´W no ano de 2003, com a obtenção simultânea de dados de temperatura, condutividade, pH, transparência e profundidade da água. Os resultados revelaram que as médias de riqueza e de diversidade específica foram maiores na zona litorânea e significativamente maiores (p24°C. Dentre os táxons observados, Desmodesmus heteracanthus (Guerr. Hentschke et Torgan, D. lunatus (W. et G. S. West Hegew. e Monoraphidium griffithii (Berk. Kom-Legn., mostraram-se indicadoras de hábitats e D. armatus (Chod. Hegew., D. denticulatus (Lagerh. An, Friedl et Hegew. e D. lunatus foram indicadoras de hidroperíodo. Este estudo demonstra a importância do hábitat e do hidroperíodo, na estrutura da comunidade de Chlorococcales s.l.This study aims to evaluate richness, density specific diversity and distribution of the Chlorococcales s.l. community in pelagic and littoral zones of different aquatic habitats (isolated lagoons, interlinked lagoons, swamps, reservoirs and streams during two hydroperiods (high and low water in a subtropical region in Southern Brazil. Phytoplankton sampling was carried out in two areas, situated within the coordinates 30(040´ - 30(010´ S and 50(030´ - 51(031´ W, in 2003 with simultaneous information on temperature, conductivity, pH, transparency and depth of water. The results showed that richness and specific diversity averages were higher in the littoral zone and significantly higher (p24°C. Among the observed taxa Desmodesmus heteracanthus (Guerr. Hentschke & Torgan, D. lunatus (W. et G. S. West Hegew. and Monoraphidium griffithii (Berk. Kom-Legn. were habitat indicators and D. armatus (Chod. Hegew., D. denticulatus (Lagerh. An, Friedl et Hegew. and D. lunatus were hydroperiod indicators. This study shows the importance of habitats and hydroperiods for Chlorococcales s.l. community structure.

  4. Monoclonal antibody to native P39 protein from Borrelia burgdorferi.

    OpenAIRE

    Sullivan, T J; Hechemy, K E; Harris, H L; Rudofsky, U H; Samsonoff, W A; Peterson, A J; Evans, B. D.; Balaban, S L

    1994-01-01

    We have produced, by using a sonicate of Borrelia burgdorferi, a monoclonal antibody (MAb), NYSP39H, that is specific for the P39 protein band. This MAb reacted with 13 isolates of B. burgdorferi but not with eight different spirochetes (four borrelias, two leptospiras, and two treponemas). Surface labeling of B. burgdorferi with biotin and subsequent treatment with Nonidet P-40 showed that P39 was not biotinylated but was extracted with Nonidet P-40, indicating that it is present within the ...

  5. Genetic population structure of the alpine species Rhododendron pseudochrysanthum sensu lato (Ericaceae inferred from chloroplast and nuclear DNA

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    Wang Wei-Kuang

    2011-04-01

    Full Text Available Abstract Background A complex of incipient species with different degrees of morphological or ecological differentiation provides an ideal model for studying species divergence. We examined the phylogeography and the evolutionary history of the Rhododendron pseudochrysanthum s. l. Results Systematic inconsistency was detected between gene genealogies of the cpDNA and nrDNA. Rooted at R. hyperythrum and R. formosana, both trees lacked reciprocal monophyly for all members of the complex. For R. pseudochrysanthum s.l., the spatial distribution of the cpDNA had a noteworthy pattern showing high genetic differentiation (FST = 0.56-0.72 between populations in the Yushan Mountain Range and populations of the other mountain ranges. Conclusion Both incomplete lineage sorting and interspecific hybridization/introgression may have contributed to the lack of monophyly among R. hyperythrum, R. formosana and R. pseudochrysanthum s.l. Independent colonizations, plus low capabilities of seed dispersal in current environments, may have resulted in the genetic differentiation between populations of different mountain ranges. At the population level, the populations of Central, and Sheishan Mountains may have undergone postglacial demographic expansion, while populations of the Yushan Mountain Range are likely to have remained stable ever since the colonization. In contrast, the single population of the Alishan Mountain Range with a fixed cpDNA haplotype may have experienced bottleneck/founder's events.

  6. PCR-based methods for the detection of L1014 kdr mutation in Anopheles culicifacies sensu lato

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    Dash Aditya P

    2009-07-01

    Full Text Available Abstract Background Anopheles culicifacies s.l., a major malaria vector in India, has developed widespread resistance to DDT and is becoming resistant to pyrethroids–the only insecticide class recommended for the impregnation of bed nets. Knock-down resistance due to a point mutation in the voltage gated sodium channel at L1014 residue (kdr is a common mechanism of resistance to DDT and pyrethroids. The selection of this resistance may pose a serious threat to the success of the pyrethroid-impregnated bed net programme. This study reports the presence of kdr mutation (L1014F in a field population of An. culicifacies s.l. and three new PCR-based methods for kdr genotyping. Methods The IIS4-IIS5 linker to IIS6 segments of the para type voltage gated sodium channel gene of DDT and pyrethroid resistant An. culicifacies s.l. population from the Surat district of India was sequenced. This revealed the presence of an A-to-T substitution at position 1014 leading to a leucine-phenylalanine mutation (L1014F in a few individuals. Three molecular methods viz. Allele Specific PCR (AS-PCR, an Amplification Refractory Mutation System (ARMS and Primer Introduced Restriction Analysis-PCR (PIRA-PCR were developed and tested for kdr genotyping. The specificity of the three assays was validated following DNA sequencing of the samples genotyped. Results The genotyping of this An. culicifacies s.l. population by the three PCR based assays provided consistent result and were in agreement with DNA sequencing result. A low frequency of the kdr allele mostly in heterozygous condition was observed in the resistant population. Frequencies of the different genotypes were in Hardy-Weinberg equilibrium. Conclusion The Leu-Phe mutation, which generates the kdr phenotype in many insects, was detected in a pyrethroid and DDT resistant An. culicifacies s.l. population. Three PCR-based methods were developed for kdr genotyping. All the three assays were specific. The ARMS method was refractory to non-specific amplification in non-stringent amplification conditions. The PIRA-PCR assay is able to detect both the codons for the phenylalanine mutation at kdr locus, i.e., TTT and TTC, in a single assay, although the latter codon was not found in the population genotyped.

  7. Escolha de cursos de pós-graduação lato sensu e os seus fatores determinantes

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    Amire Tauil

    2015-09-01

    Full Text Available The professionalization of the educational branch makes HEIs (Higher Education Institutions seek information on attractiveness of their courses, that is, which factors are decisive for the choice of this institution. Thus, this study aims to identify the determining factors in the choice of students for post-graduation courses in a higher education institution. To meet the objective of a descriptive research with a quantitative approach was carried out in a higher education institution of Vitória,Espirito Santo, through the application of a questionnaire to 210 students in their first day of class in the graduate school. Data were analyzed using descriptive and multivariate statistics. The main results, which contribute positively to the attractiveness of the HEIs, are related to factors such as: the structure of the higher education institution, for example, campus security and the place of meal; update of the course, teachers with experience in the labor market and updating course subjects; and personal reasons, such as the location of the institution. These factors can be an important indicator for HEIs offering postgraduate course.

  8. [Relationship between characteristics of sexual behavior and male sperm competitive ability in taxa of superspecies complex Mus musculus sensu lato].

    Science.gov (United States)

    Ambaryan, A B; Maltzev, A N; Kotenkova, E V

    2015-01-01

    Some physiological parameters that determine quality of male sperm (its concentration, spermatozoa morphology) and testicle size vary in integrity, i.e. the bigger are testicles the higher is sperm quality. Therefore, the estimate of testicles relative mass is often used as a characteristic of sperm competitive ability when comparing phylogenetically close mammal species. In house mice belonging to the superspecies complex Mus musculus s.l., testicles relative mass is greater in exoanthropic species than in synanthropic ones. It is shown in our study that this pattern is apparent also at the intraspecies level since testicles mass index, sperm concentration, and percentage of morphologically normal spermatozoa in subspecies Mus musculus wagneri, which is facultatively synanthropic, are higher compared with synanthropic subspecies M m. musculus. An analysis of sexual behavior of the three forms (namely, exoanthropic species M. spicilegus and two subspecies mentioned above) indicates that in M. spicilegus both sexual behavior efficiency and ejaculation rate during coupling were higher as compared with other two subspecies. Based on the analysis of life pattern, reproduction systems, and group spatial-ethological structure, the hypotheses are formulated that explain the maintenance of selection directed to increase of sperm competitive ability in exoanthropic house mice species.

  9. Borniopsis mortoni sp. n. (Heterodonta, Galeommatoidea, Galeommatidae sensu lato), a new bivalve commensal with a synaptid sea cucumber from Japan

    Science.gov (United States)

    Goto, Ryutaro; Ishikawa, Hiroshi

    2016-01-01

    Abstract The Galeommatoidea is a bivalve superfamily that exhibits high species diversity in shallow waters. Many members of this superfamily are associated commensally with burrowing marine invertebrates in benthic sediments. The genus Borniopsis is known only from eastern Asia and exhibits high host diversity (e.g., mantis shrimps, crabs, holothurians, sipunculans and echiurans). A new species, Borniopsis mortoni sp. n., is described from mud flats at the mouth of the Souzu River, southwestern Shikoku Island, Japan. This species has elongate-ovate shells covered by a tan to dark brown periostracum, and lives attached by both its foot and byssal threads to the body surface of the synaptid sea cucumber Patinapta ooplax. Several individuals of Borniopsis mortoni are often found on the same host, but sometimes more than 10 individuals can occur together. Borniopsis mortoni is one of the smallest species in this genus. Probably, its small body size is an adaptation to the mode of life in a narrow host burrow. Until now, only two other Borniopsis species were known to have commensal associations with synaptids. Thus, this is the third example of a synaptid-associated species from this genus. In addition, we briefly review the galeommatoideans commensal with apodid sea cucumbers. PMID:27667939

  10. Multilocus microsatellite typing (MLMT of strains from Turkey and Cyprus reveals a novel monophyletic L. donovani sensu lato group.

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    Evi Gouzelou

    Full Text Available BACKGROUND: New foci of human CL caused by strains of the Leishmania donovani (L. donovani complex have been recently described in Cyprus and the Çukurova region in Turkey (L. infantum situated 150 km north of Cyprus. Cypriot strains were typed by Multilocus Enzyme Electrophoresis (MLEE using the Montpellier (MON system as L. donovani zymodeme MON-37. However, multilocus microsatellite typing (MLMT has shown that this zymodeme is paraphyletic; composed of distantly related genetic subgroups of different geographical origin. Consequently the origin of the Cypriot strains remained enigmatic. METHODOLOGY/PRINCIPAL FINDINGS: The Cypriot strains were compared with a set of Turkish isolates obtained from a CL patient and sand fly vectors in south-east Turkey (Çukurova region; CUK strains and from a VL patient in the south-west (Kuşadasi; EP59 strain. These Turkish strains were initially analyzed using the K26-PCR assay that discriminates MON-1 strains by their amplicon size. In line with previous DNA-based data, the strains were inferred to the L. donovani complex and characterized as non MON-1. For these strains MLEE typing revealed two novel zymodemes; L. donovani MON-309 (CUK strains and MON-308 (EP59. A population genetic analysis of the Turkish isolates was performed using 14 hyper-variable microsatellite loci. The genotypic profiles of 68 previously analyzed L. donovani complex strains from major endemic regions were included for comparison. Population structures were inferred by combination of bayesian model-based and distance-based approaches. MLMT placed the Turkish and Cypriot strains in a subclade of a newly discovered, genetically distinct L. infantum monophyletic group, suggesting that the Cypriot strains may originate from Turkey. CONCLUSION: The discovery of a genetically distinct L. infantum monophyletic group in the south-eastern Mediterranean stresses the importance of species genetic characterization towards better understanding, monitoring and controlling the spread of leishmaniasis in this region.

  11. The fauna of the family Bombycidae sensu lato (Insecta, Lepidoptera, Bombycoidea) from Mainland China, Taiwan and Hainan Islands.

    Science.gov (United States)

    Wang, Xing; Wang, Min; Zolotuhin, Vadim V; Hirowatari, Toshiya; Wu, Shipher; Huang, Guo-Hua

    2015-07-22

    Seventy-seven species of family Bombycidae s. lat., belonging to 25 genera in three subfamilies, that have been recorded from China are listed and described, with illustrations of the adults, preimaginal stages (if available), and their genitalia. Keys to subfamilies and genera are provided. Two new genera and four new species are described, two subgenera are raised to generic status, seven new combinations are made, and one genus and six species are newly recorded from China. The new taxa are as follows: Rotunda Wang, X. & Zolotuhin, gen. nov., Comparmustilia Wang, X. & Zolotuhin, gen. nov., Triuncina daii Wang, X. & Zolotuhin, sp. nov., Triuncina xiongi Wang, X. & Zolotuhin, sp. nov., Gnathocinara boi Wang, X. & Zolotuhin, sp. nov. and Promustilia yajiangensis Wang, X. & Zolotuhin, sp. nov. The taxa newly recorded for China are: Sesquiluna Forbes, 1955; Trilocha friedeli Dierl, 1978; Bivincula kalikotei Dierl, 1978; Sesquiluna forbesi Zolotuhin & Witt, 2009; Mustilizans lepusa Zolotuhin, 2007; Smerkata brechlini (Zolotuhin, 2007) and Mustilia castanea Moore, 1879. The seven new combinations are: Rotunda rotundapex (Miyata & Kishida, 1990), comb. nov., Triuncina nitida (Chu & Wang, L.Y., 1993), comb. nov., Gunda sesostris (Vuillot, 1893), comb. nov., Smerkata fusca (Kishida, 1993), comb. nov., Comparmustilia sphingiformis (Moore, 1879), comb. nov., Comparmustilia semiravida (Yang, 1995), comb. nov., Comparmustilia gerontica (West, 1932), comb. nov.. The two subgenera raised to generic level are: Promustilia Zolotuhin, 2007, stat. nov. and Smerkata Zolotuhin, 2007, stat. nov.. The distributions of the species in China were determined and distributional maps provided. All type specimens of the new species described here are deposited in the College of Plant Protection, Hunan Agricultural University, China (HUNAU); Department of Entomology, South China Agricultural University, China (SCAU); Kyushu University Museum, Kyushu University, Japan (KUM), and Entomological Museum Thomas J. Witt, Munich, Germany (MWM).

  12. Brazilian Atlantic Forest lato sensu: the most ancient Brazilian forest, and a biodiversity hotspot, is highly threatened by climate change

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    AF. Colombo

    Full Text Available After 500 years of exploitation and destruction, the Brazilian Atlantic Forest has been reduced to less the 8% of its original cover, and climate change may pose a new threat to the remnants of this biodiversity hotspot. In this study we used modelling techniques to determine present and future geographical distribution of 38 species of trees that are typical of the Brazilian Atlantic Forest (Mata Atlântica, considering two global warming scenarios. The optimistic scenario, based in a 0.5% increase in the concentration of CO2 in the atmosphere, predicts an increase of up to 2 °C in the Earth's average temperature; in the pessimistic scenario, based on a 1% increase in the concentration of CO2 in the atmosphere, temperature increase may reach 4 °C. Using these parameters, the occurrence points of the studied species registered in literature, the Genetic Algorithm for Rule-set Predictions/GARP and Maximum entropy modeling of species geographic distributions/MaxEnt we developed models of present and future possible occurrence of each species, considering Earth's mean temperature by 2050 with the optimistic and the pessimistic scenarios of CO2 emission. The results obtained show an alarming reduction in the area of possible occurrence of the species studied, as well as a shift towards southern areas of Brazil. Using GARP, on average, in the optimistic scenario this reduction is of 25% while in the pessimistic scenario it reaches 50%, and the species that will suffer the worst reduction in their possible area of occurrence are: Euterpe edulis, Mollinedia schottiana, Virola bicuhyba, Inga sessilis and Vochysia magnifica. Using MaxEnt, on average, in the optimistic scenario the reduction will be of 20% while in the pessimistic scenario it reaches 30%, and the species that will suffer the worst reduction are: Hyeronima alchorneoides, Schefflera angustissima, Andira fraxinifolia and the species of Myrtaceae studied.

  13. Comparative analysis of spermatids of Rhipicephalus sanguineus sensu lato (Ixodidae) and Ornithodoros rostratus ticks (Argasidae): morphophysiology aimed at systematics.

    Science.gov (United States)

    Sampieri, Bruno Rodrigues; Calligaris, Izabela Bragião; Matos, Renata da Silva; Páez, Fredy Arvey Rivera; Bueno, Odair Corrêa; Camargo-Mathias, Maria Izabel

    2016-02-01

    The phylogenetic relationships among tick species (Acari: Ixodida) have been revisited by several researchers over the last decades. Two subfamilies, Rhipicephalinae (Ixodidae) and Ornithodorinae (Argasidae), deserve special attention. The male reproductive system morphology, as well as the ultrastructure of the germ cells, may provide important information for phylogeny and systematics of metazoan groups, with spermatozoa exhibiting characters that can be used for this purpose. With that information in mind, this study aimed at evaluating, through a comparative analysis, the morphology of the male reproductive systems and germ cells of ticks species Rhipicephalus sanguineus and Ornithodoros rostratus. In order to do that, histology and scanning electron microscopy techniques were used. The results have shown that despite the similarities in the general morphology of the male reproductive system among studied Ixodida so far, there are morphological differences among the species studied herein, mainly the U-shaped testis (ancestral character) in O. rostratus and the pair testes (derived character) in R. sanguineus, and the general morphology of germ cells (spermatids V). Besides that, the morphological changes observed during the spermiogenesis appear to be different between the species studied here, probably characterizing the two families considered. The data generated in this study showed the importance of comparative internal morphology studies, mainly in regard to spermatology, despite the morphological data obtained herein not being enough to product a cladogram (sperm cladistics), it was already possible to observe clear differences among families Argasidae and Ixodidae in regard to the organization of their male reproductive systems and concerning the external morphology of spermatids. Data yet to be obtained through transmission electron microscopy techniques will allow the application of spermiocladistics and spermiotaxonomy as tools for tick systematics.

  14. Annea gen. nov.(Detarieae, Caesalpinioideae, Leguminosae): a home for two species long misplaced in Hymenostegia sensu lato. Phytotaxa

    NARCIS (Netherlands)

    Mackinder, B.A.; Wieringa, J.J.

    2013-01-01

    A new genus Annea is described to accommodate two tropical African legume species previously misplaced in Hymenostegia (Leguminosae, Caesalpinioideae, Detarieae). Annea gen. nov. is widespread in tropical Africa but has an unusual disjunct generic distribution, occurring in both upper and lower Guin

  15. Borniopsis mortoni sp. n. (Heterodonta, Galeommatoidea, Galeommatidae sensu lato, a new bivalve commensal with a synaptid sea cucumber from Japan

    Directory of Open Access Journals (Sweden)

    Ryutaro Goto

    2016-09-01

    Full Text Available The Galeommatoidea is a bivalve superfamily that exhibits high species diversity in shallow waters. Many members of this superfamily are associated commensally with burrowing marine invertebrates in benthic sediments. The genus Borniopsis is known only from eastern Asia and exhibits high host diversity (e.g., mantis shrimps, crabs, holothurians, sipunculans and echiurans. A new species, Borniopsis mortoni sp. n., is described from mud flats at the mouth of the Souzu River, southwestern Shikoku Island, Japan. This species has elongate-ovate shells covered by a tan to dark brown periostracum, and lives attached by both its foot and byssal threads to the body surface of the synaptid sea cucumber Patinapta ooplax. Several individuals of B. mortoni are often found on the same host, but sometimes more than 10 individuals can occur together. Borniopsis mortoni is one of the smallest species in this genus. Probably, its small body size is an adaptation to the mode of life in a narrow host burrow. Until now, only two other Borniopsis species were known to have commensal associations with synaptids. Thus, this is the third example of a synaptid-associated species from this genus. In addition, we briefly review the galeommatoideans commensal with apodid sea cucumbers.

  16. A Molecular Phylogeny of Anopheles Annulipes (Diptera: Culicidae) sensu lato: The Most Species-Rich Anopheline Complex

    Science.gov (United States)

    2006-09-27

    Springsc 130°58E 12°42S K-14[3],N-17 N-17(91) DQ420404 DQ420596 DQ420468 DQ420532 42 Jim Jim Ckd 133°05E 13°05S K-14[3],N-17, K-14(63) DQ420396...of An. annulipes s.l. may be the cause of much of the diYculty that taxonomists have had in explaining the extensive morphological variation within

  17. Effectiveness of Beauveria bassiana sensu lato strains for biological control against Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in China.

    Science.gov (United States)

    Sun, Ming; Ren, Qiaoyun; Guan, Guiquan; Li, Yufeng; Han, Xueqing; Ma, Chao; Yin, Hong; Luo, Jianxun

    2013-10-01

    Owing to the need to combat the spread of acaricide-resistant ticks, the development of long-term biological control has become a hot topic for tick control. In this study, we investigated the pathogenicity of three Beauveria bassiana isolates on the engorged female Rhipicephalus (Boophilus) microplus ticks using different conidial concentrations. The results showed that B. bassiana B.bAT17 was highly pathogenic against engorged R. (B.) microplus females, resulting in lethal time (LT50 and LT90) of 7.14 and 9.33 days at a concentration of 10(9)conidia/ml. R. (B.) microplus females treated with B. bassiana B.bAT17 significantly reduced the amount of ovipositioning; and most ticks died before they could begin to oviposit. Proteases and chitinases were analyzed in order to establish a screening method for identification of high virulent strains. This study has confirmed the significant pathogenic effect of entomopathogenic fungi against engorged R. (B.) microplus females in China, and further studies on the efficiency of the fungus against ticks in the field are required.

  18. Growth inhibiting activity of lipophilic extracts from Dipsacus sylvestris Huds. roots against Borrelia burgdorferi s. s. in vitro.

    Science.gov (United States)

    Liebold, T; Straubinger, R K; Rauwald, H W

    2011-08-01

    Fresh first year roots from Dipsacus sylvestris HUDS. were extracted with 70% ethanol, ethyl acetate as well as dichloromethane. Extracts were solubilized in water (lipophilic extracts with addition of polysorbate 80) and tested for their activity against Borrelia burgdorferi sensu stricto in vitro during an eight-day period using amoxicillin as standard. The hydroethanolic extract showed no growth inhibition whereas significant growth inhibiting activity could be shown in the two less polar fractions for the first time. Strongest inhibition was found in the ethyl acetate extract. The effect of polysorbate 80 on bacterial growth was examined and found to be negligible. As the nature of bioactive constituents has not been clarified yet, a micellar electrokinetic capillary chromatography fingerprint analysis for a methanolic extract was applied including loganin, chlorogenic acid, cantleyoside and caffeic acid as marker substances.

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

    Directory of Open Access Journals (Sweden)

    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

  20. Molecular characterization of Echinococcus granulosus sensu stricto and Echinococcus canadensis in humans and livestock from Algeria.

    Science.gov (United States)

    Zait, Houria; Kouidri, Mokhtaria; Grenouillet, Florence Elisabeth; Umhang, Gérald; Millon, Laurence; Hamrioui, Boussad; Grenouillet, Frédéric

    2016-06-01

    In Algeria, previous studies investigated genotypes of Echinococcus granulosus sensu lato in animals and identified E. granulosus sensu stricto (s.s.) genotypes G1 and G3 whereas Echinococcus canadensis genotype G6 was only reported from dromedary cysts. Molecular data on human cystic echinococcosis (CE) were limited. We implemented a large genotyping study of hydatid cysts from humans and livestock animals to specify CE's molecular epidemiology and the genetic diversity in Algeria. Fifty-four human CE cysts from patients predominantly admitted in surgical units from Mustapha Hospital, Algiers, and 16 cysts from livestock animals gathered in two geographically distinct slaughterhouses, Tiaret and Tamanrasset, were collected. Molecular characterization was performed using sequencing of two mitochondrial genes, cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit I (NDI). In humans, G1 of E. granulosus s.s. was the main genotype (90.7 %); four samples (7.4 %) were characterized as E. granulosus s.s. G3 and one cyst as E. canadensis G6 (1.8 %). This molecular confirmation of E. canadensis G6 human infection in Algeria was observed in a Tuareg female living in a desertic area in Tamanrasset. All cysts from sheep, cattle, and goat were identified as E. granulosus s.s. G1 and the two cysts originating from dromedary as E. canadensis G6. Twenty concatenated haplotypes (COI + NDI) were characterized. Among E. granulosus s.s., one haplotype (HL1) was highly predominant in both humans and animals cysts (71.6 %). This study revealed main occurrence of E. granulosus s.s. in humans and livestock animals, with description of a predominant shared haplotype corresponding to the main worldwide observed haplotype E.granulosus s.s. G1. E. canadensis G6 was limited to South Algeria, in dromedary as well as in human.

  1. Autophagy suppresses host adaptive immune responses toward Borrelia burgdorferi

    NARCIS (Netherlands)

    Buffen, Kathrin; Oosting, Marije; Li, Yang; Kanneganti, Thirumala-Devi; Netea, Mihai G.; Joosten, Leo A. B.

    2016-01-01

    Inhibition of autophagy increases the severity of murine Lyme arthritis and human adaptive immune responses against B. burgdorferi. We have previously demonstrated that inhibition of autophagy increased the Borrelia burgdorferi induced innate cytokine production in vitro, but little is known regardi

  2. Reprint of "Echinococcus granulosus sensu stricto (s.s.) from the critically endangered antelope Addax nasomaculatus in Tunisia".

    Science.gov (United States)

    Boufana, Belgees; Saïd, Yousra; Dhibi, Mokhtar; Craig, Philip S; Lahmar, Samia

    2017-01-01

    Echinococcus granulosus sensu lato (s.l.) is a zoonotic disease highly endemic in Tunisia. Canids including stray and semi-stray dogs, jackals and foxes are known as definitive hosts and a wide range of ungulates have been shown to harbour the metacestode hydatid stage and may serve as intermediate hosts. Fertile hydatid cysts of Echinococcus equinus and E. granulosus sensu stricto (s.s.) were recently molecularly identified for the first time from Tunisian donkeys. E. granulosus (s.s.) was also identified from wild boars in Tunisia. Here we report the confirmation of hydatid cysts caused by E. granulosus (s.s.) in the critically endangered antelope, Addax nasomaculatus in Tunisia. DNA-based molecular analysis revealed that A. nasomaculatus was infected with E. granulosus (s.s.) which had a 100% identity with the main globally distributed E. granulosus (s.s.) (EgTu01) haplotype. Cysts of Taenia hydatigena (n=33) were also observed on the liver and in the body cavity. Due to their endangered status and their relatively small numbers, it is unlikely that hydatid infection of A. nasomaculatus will form a major contribution to the epidemiology and transmission of E. granulosus in Tunisia, but infection may result in pathology, morbidity and early mortality, and may still play a role in the perpetuation of the parasite in wildlife cycles.

  3. Lyme borreliosis vaccination: the facts, the challenge and the future

    NARCIS (Netherlands)

    T.J. Schuijt; J.W. Hovius; T. van der Poll; A.P. van Dam; E. Fikrig

    2011-01-01

    Lyme disease, or Lyme borreliosis, the most prevalent arthropod-borne disease in the Western world, is caused by spirochetes belonging to the Borrelia burgdorferi sensu lato group and is predominantly transmitted through Ixodes ticks. There is currently no vaccine available to prevent Lyme borrelios

  4. Tired of Lyme borreliosis: Lyme borreliosis in the Netherlands

    NARCIS (Netherlands)

    J. Coumou; T. van der Poll; P. Speelman; J.W.R. Hovius

    2011-01-01

    Lyme borreliosis has become the most common vector-borne illness in North Eastern USA and Europe. It is a zoonotic disease, with well-defined symptoms, caused by B. burgdorferi sensu lato, and transmitted by ticks. Lyme borreliosis is endemic in the Netherlands with a yearly incidence of approximate

  5. Sensitivity of a point of care tick-test for the development of Lyme borreliosis

    NARCIS (Netherlands)

    Sprong, H.; Leeuwen, van A.D.; Fonville, M.; Harms, M.; Vliet, van A.J.H.; Pelt, van W.; Ferreira, J.A.; Wijngaard, van den C.C.

    2013-01-01

    Background: A commercially available self-test for the detection of Borrelia burgdorferi sensu lato in ticks was evaluated for its ability to predict erythema migrans formation. Findings: The self-test was performed on 127 Ixodes ricinus from 122 humans that reported tick bites at enrolment and occu

  6. Dissecting Lyme borreliosis; Clinical aspects, pathogenesis and prevention

    NARCIS (Netherlands)

    J. Coumou

    2016-01-01

    Lyme borreliosis (LB) is the most prevalent vector-borne disease in Western Europe and Northeastern parts of the USA. The causative agents of LB are spirochetes belonging to the Borrelia burgdorferi sensu lato group, which are transmitted by Ixodes ticks. Since the late 1970’s, researchers in the Un

  7. 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).

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2011-02-01

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

  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

    Directory of Open Access Journals (Sweden)

    Tsachev Ilia

    2007-01-01

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

  11. Proteome Analysis of Borrelia burgdorferi Response to Environmental Change

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-11-02

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

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

    Science.gov (United States)

    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

  13. The Text of Aristotle's De Sensu and De Memoria

    DEFF Research Database (Denmark)

    Bloch, David Kristian

    2008-01-01

    An examination of alle the manuscripts of the period 10th-14th centuries, which contain the Aristotelian treatises De Sensu and De Memoria. The article establishes a stemma codicum that includes alle these manuscripts.......An examination of alle the manuscripts of the period 10th-14th centuries, which contain the Aristotelian treatises De Sensu and De Memoria. The article establishes a stemma codicum that includes alle these manuscripts....

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

    Directory of Open Access Journals (Sweden)

    Eva Sapi

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

  15. Evidence supporting the presence of Borrelia burgdorferi in Missouri.

    Science.gov (United States)

    Feir, D; Santanello, C R; Li, B W; Xie, C S; Masters, E; Marconi, R; Weil, G

    1994-10-01

    Although Lyme disease is commonly seen in the southcentral United States, the epidemiology of the disease is poorly defined there. The purpose of this study was to document the presence of Borrelia burgdorferi in ticks collected in southeastern Missouri and around the city of St. Louis. Spirochetes were detected and identified as B. burgdorferi by immunofluorescent antibody (IFA) tests using the monoclonal antibody H5332 in 1.9% of Amblyomma americanum and 2.0% of Dermacentor variabilis ticks collected. The identity of IFA-positive organisms was verified by polymerase chain reactions (PCRs) with two different sets of B. burgdorferi-specific primers followed by Southern blotting. The DNA sequences of amplified 371-basepair PCR products from two positive Missouri ticks showed 97-98% identity with that obtained by the same method for the B31 strain of B. burgdorferi. These results confirm that B. burgdorferi is present in questing D. variabilis and A. americanum ticks in areas of Missouri where Lyme disease occurs. Additional studies are needed to determine the role of these ticks in the epidemiology of Lyme disease in Missouri and neighboring states.

  16. Echinococcus ortleppi (G5) and Echinococcus granulosus sensu stricto (G1) loads in cattle from Southern Brazil.

    Science.gov (United States)

    Balbinotti, Helier; Santos, Guilherme B; Badaraco, Jeferson; Arend, Ana C; Graichen, Daniel Ângelo S; Haag, Karen L; Zaha, Arnaldo

    2012-09-10

    Echinococcus granulosus sensu stricto (G1) and Echinococcus ortleppi (G5) are haplotypes of the parasite formerly known as Echinococcus granulosus sensu lato, which in its larval stage causes cystic hydatid disease, endemic in Southern Brazil. Epidemiological and molecular knowledge about the haplotypes occurring in a region is essential to control the spread of the disease. The aim of this work was to analyze the haplotype frequency and fertility of hydatid cysts in cattle from the state of Rio Grande do Sul. Cysts were collected and classified according to their fertility status. DNA was extracted from protoscoleces and germinal layers and then used as template for the amplification of the cytochrome c oxidase subunit 1 gene by PCR. Amplicons were purified and sequenced, and the sequences were analyzed for haplotype identification. A total of 638 fertile cysts collected in the last ten years were genotyped. On average, G1 (56.6%) was more frequent than G5 (43.4%). In lungs, the G5 haplotype exhibited a higher parasite load (52.8%), whereas in the liver, G1 was more frequent (90.4%). The analysis revealed an increase in the frequency of G5 haplotype cysts during the period of sampling, and an increase in the abundance of fertile cysts has also been observed in the last several years. Most infertile cysts were genotyped as G1. The possible factors involved in the increase in the proportion of E. ortleppi (G5) and the consequences of this increase are discussed. This study suggests that the proportion of E. ortleppi (G5) loads in cattle may be increasing overtime.

  17. Canine echinococcosis: genetic diversity of Echinococcus granulosus sensu stricto (s.s.) from definitive hosts.

    Science.gov (United States)

    Boufana, B; Lett, W; Lahmar, S; Griffiths, A; Jenkins, D J; Buishi, I; Engliez, S A; Alrefadi, M A; Eljaki, A A; Elmestiri, F M; Reyes, M M; Pointing, S; Al-Hindi, A; Torgerson, P R; Okamoto, M; Craig, P S

    2015-11-01

    Canids, particularly dogs, constitute the major source of cystic echinococcosis (CE) infection to humans, with the majority of cases being caused by Echinococcus granulosus (G1 genotype). Canine echinococcosis is an asymptomatic disease caused by adult tapeworms of E. granulosus sensu lato (s.l.). Information on the population structure and genetic variation of adult E. granulosus is limited. Using sequenced data of the mitochondrial cytochrome c oxidase subunit 1 (cox1) we examined the genetic diversity and population structure of adult tapeworms of E. granulosus (G1 genotype) from canid definitive hosts originating from various geographical regions and compared it to that reported for the larval metacestode stage from sheep and human hosts. Echinococcus granulosus (s.s) was identified from adult tapeworm isolates from Kenya, Libya, Tunisia, Australia, China, Kazakhstan, United Kingdom and Peru, including the first known molecular confirmation from Gaza and the Falkland Islands. Haplotype analysis showed a star-shaped network with a centrally positioned common haplotype previously described for the metacestode stage from sheep and humans, and the neutrality indices indicated population expansion. Low Fst values suggested that populations of adult E. granulosus were not genetically differentiated. Haplotype and nucleotide diversities for E. granulosus isolates from sheep and human origin were twice as high as those reported from canid hosts. This may be related to self-fertilization of E. granulosus and/or to the longevity of the parasite in the respective intermediate and definitive hosts. Improved nuclear single loci are required to investigate the discrepancies in genetic variation seen in this study.

  18. Minimal-Change Disease Secondary to Borrelia burgdorferi Infection

    Directory of Open Access Journals (Sweden)

    Ewa Kwiatkowska

    2012-01-01

    Full Text Available 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. (1997. In available literature, there were reports of human's glomerulonephritis associated with Borrelia burgdorferi infection. These cases refer to membranous and mesangial proliferative glomerulonephritis (Kirmizis and Chatzidimitriou (2010, Zachäus (2008, and Kirmizis et al. (2004. In this paper, we present the case of minimal-change disease (MCD as a result of Borrelia burgdorferi infection.

  19. Minimal-Change Disease Secondary to Borrelia burgdorferi Infection.

    Science.gov (United States)

    Kwiatkowska, Ewa; Gołembiewska, Edyta; Ciechanowski, Kazimierz; Kędzierska, Karolina

    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. (1997)). In available literature, there were reports of human's glomerulonephritis associated with Borrelia burgdorferi infection. These cases refer to membranous and mesangial proliferative glomerulonephritis (Kirmizis and Chatzidimitriou (2010), Zachäus (2008), and Kirmizis et al. (2004)). In this paper, we present the case of minimal-change disease (MCD) as a result of Borrelia burgdorferi infection.

  20. [Nucleotide sequence analysis of a species specific probe by an inserted fragment from recombinant plasmid pCX7 of L. interrogans sensu stricto serovar lai].

    Science.gov (United States)

    Dai, B; Xiao, J; Yan, Z; Shen, C; Li, S; Fang, Z

    1998-12-01

    The etiological agents of leptospirosis are the pathogenic leptospires (L. interrogans sensu lato) which can be divided into 223 serovars organized into 23 serogroups. The serovar remains the basic taxon, but serotyping may now be accomplished and recognized by acceptable methods. Complementary molecular approaches are being used extensively to assess genetic relatedness amongst leptospires with restriction endonuclese analysis (REA), pulse field gel electrophoresis (PFGE) and DNA-DNA hybridization as well established tools. However, the method is cumbersome and unsuitable for routine application. To develop a sensitive and specific method for identification of pathogenic leptospires, a genomic library of L. interrogans sensu stricto serovar lai was constructed with the plasmid vector pUC9. A recombinant plasmid, designated pCX7 which has homologous fragment of pathogenic leptospires was screened from the bank. pCX7 could recognize pathogenic leptospiral DNA fragment 1.7 kb of strain 017 without cross hybridization to nonpathogenic leptospiral DNA. Inserted fragment of pCX7 DNA sequencing was performed by Dr. Yan Zhengxin (Max-Plank-Institut fur Biology, Tubingen, Germany). Insert fragment was cloned into pBluescript and sequenced by using ABI(Applied Bio. Systems, Model 373A). Nucleotide sequences were analyzed by Dr. Xiao Jianguo (Texas University Medical School and School of Public Health, Center for Infectious Diseases) using a suit of computer program (NIH). One open reading frame of 306 nucleotids were identified. There were identifiable initiation codons, terminators, pribnow box and sextama box within the sequenced regions. These results further confirmed that the little homology between L. interrogans sensu strito and L. borgpeterseni serovar javanica, L. inadai serovar ranarun and serovar manhao (L. genomospecies 2), L. biflexa serovar patoc, L. illini. pCX7 DNA probe could provide a base for identification and classification of leptospires.

  1. The phylogeny and taxonomic status of the Chlorocystini (sensu stricto) (Homoptera, Tibicinidae)

    NARCIS (Netherlands)

    Boer, de A.J.

    1995-01-01

    The “Baeturia and related genera complex”, as defined earlier (De Boer, 1990) by shared aedeagal characters, is identified as the tribe Chlorocystini (sensu stricto). The Prasiini (sensu stricto) are identified as the sister group of the Chlorocystini (sensu stricto), while the genus Muda is recogni

  2. 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.

  3. Lipid exchange between Borrelia burgdorferi and host cells.

    Directory of Open Access Journals (Sweden)

    Jameson T Crowley

    2013-01-01

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

  4. Minimal-Change Disease Secondary to Borrelia burgdorferi Infection

    OpenAIRE

    Ewa Kwiatkowska; Edyta Gołembiewska; Kazimierz Ciechanowski; Karolina Kędzierska

    2012-01-01

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

  5. Screening of cytoplasmic DNA diversity between and within Lupinus mutabilis Sweet and Lupinus albus sensu lato by restriction fragment length polymorphism (RFLP).

    Science.gov (United States)

    Olczak, T; Rurek, M; Jańska, H; Augustyniak, H; Sawicka-Sienkiewicz, E J

    2001-01-01

    Seven populations and five mutant lines of the Andean lupin and four species from the section Albus were screened for their mitochondrial and chloroplast polymorphisms. For this purpose the RFLP method with EcoRI as a restriction enzyme was used. Lupinus luteus, Lupinus albus and Phaseolus vulgaris organellar clones as well as amplified fragments were used as probes. We found that mitochondrial probes were more suitable than chloroplast probes for identification of inter- and intra-specific variations within the examined material. Most mitochondrial probes differentiate the two species investigated. A high level of mitochondrial polymorphism was observed among the populations of L. mutabilis in contrast to monomorphism among the species in the section Albus. A limited polymorphism was detected between the mutant lines of L. mutabilis. We conclude from this study that the mitochondrial RFLP analysis is a valuable tool for identification of variability among Andean lupin populations.

  6. Analysis of the population genetics of Opisthorchis viverrini sensu lato in the Nam Ngum River wetland, Lao PDR, by multilocus enzyme electrophoresis.

    Science.gov (United States)

    Kiatsopit, Nadda; Sithithaworn, Paiboon; Saijuntha, Weerachai; Pitaksakulrat, Opal; Petney, Trevor N; Webster, Joanne P; Andrews, Ross H

    2014-08-01

    A previous population genetics study of Opisthorchis viverrini from a locality in an endemic area in Thailand found little genetic variation over time and second intermediate fish host species. Since a similar comparative analysis is not available for Lao PDR, we conducted a study of O. viverrini from different endemic foci in Vientiane Province, Lao PDR, based on spatial, temporal and fish host species. A total of 620 adult O. viverrini originating from the Nam Ngum River wetland were analysed at five previously defined polymorphic enzyme loci. Of these worms, 252 were from six different localities (spatial samples), 162 worms from different years (temporal samples) and 206 worms from four different cyprinid fish species. Significant heterozygote deficiency was found in most O. viverrini populations with levels of genetic differentiation ranging between F ST 0.0000 and 0.0197 suggesting that gene flow occurred at a variable rate. The role of temporal factors and fish host species had little influence on the level of genetic differentiation. As for O. viverrini from Thailand, these findings indicate that self-fertilization and/or a clonal distribution of O. viverrini occurs in Lao PDR. Unlike the results for O. viverrini from Thailand, spatial population substructuring may be the underlying population processes for O. viverrini in Lao PDR. These findings indicate that geographical variation may contribute to the transmission dynamics of the parasite with implications for parasite control. However, other host factors, such as snail intermediate hosts and mammal reservoir hosts, as well as human beings, may also play significant roles.

  7. Co-occurrence and distribution of East (L1014S) and West (L1014F) African knock-down resistance in Anopheles gambiae sensu lato population of Tanzania

    Science.gov (United States)

    Kabula, Bilali; Kisinza, William; Tungu, Patrick; Ndege, Chacha; Batengana, Benard; Kollo, Douglas; Malima, Robert; Kafuko, Jessica; Mohamed, Mahdi; Magesa, Stephen

    2014-01-01

    Objective Insecticide resistance molecular markers can provide sensitive indicators of resistance development in Anopheles vector populations. Assaying these makers is of paramount importance in the resistance monitoring programme. We investigated the presence and distribution of knock-down resistance (kdr) mutations in Anopheles gambiae s.l. in Tanzania. Methods Indoor-resting Anopheles mosquitoes were collected from 10 sites and tested for insecticide resistance using the standard WHO protocol. Polymerase chain reaction-based molecular diagnostics were used to genotype mosquitoes and detect kdr mutations. Results The An. gambiae tested were resistance to lambdacyhalothrin in Muheza, Arumeru and Muleba. Out of 350 An. gambiae s.l. genotyped, 35% were An. gambiae s.s. and 65% An. arabiensis. L1014S and L1014F mutations were detected in both An. gambiae s.s. and An. arabiensis. L1014S point mutation was found at the allelic frequency of 4–33%, while L1014F was at the allelic frequency 6–41%. The L1014S mutation was much associated with An. gambiae s.s. (χ2 = 23.41; P mutaciones de resistencia knockdown (kdr) en Anopheles gambiae s.l. en Tanzania. Métodos Se recolectaron mosquitos Anopheles intradomiciliarios de 10 lugares diferentes y se evaluaron en busca de resistencia a insecticidas utilizando el protocolo estándar de la OMS. Mediante un diagnóstico molecular basado en la PCR se genotiparon los mosquitos y se detectaron los genotipos kdr. Resultados Los An. gambiae evaluados eran resistentes a lambdacialotrina en Muheza, Arumeru y Muleba. De 350 An. gambiae s.l. genotipados, 35% eran An. gambiae s.s. y 65% eran An. arabiensis. Se detectaron mutaciones L1014S y L1014F tanto en An. gambiae s.s. como en An. arabiensis. La mutación puntual L1014S se encontró con una frecuencia alélica de 4-33%, mientras que L1014F tenía una frecuencia alélica de 6-14%. La mutación L1014S estaba ampliamente asociada a An. gambiae s.s. (Chi-Cuadrado = 23.41; P mutaciones de L1014S y L1014F junto con informes de resistencia a los insecticidas sugiere que la resistencia a piretroides se está convirtiendo en un fenómeno común entre las poblaciones del vector de la malaria en Tanzania. La presencia de la mutación L1014F en estas poblaciones del Este de África indican la diseminación del gen a lo largo del continente africano. Determinar las implicaciones potenciales a nivel operativo de estos hallazgos sobre el control de la malaria requiere de más estudios. PMID:24386946

  8. Genetic diversity of Fusarium graminearum sensu lato isolates from wheat associated with Fusarium Head Blight in diverse geographic locations of Argentina.

    Science.gov (United States)

    Consolo, Verónica F; Ortega, Leonel M; Salerno, Graciela; Astoreca, Andrea L; Alconada, Teresa M

    2015-01-01

    Fusarium Head Blight is an important wheat disease in the Argentine Pampas region, being Fusarium graminearum the predominant pathogen. DNA polymorphism of the isolates was analyzed by IGS-RFLP and ISSR. IGS-RFLP and ISSR profiling were carried out using six endonucleases and eight primers, respectively. IGS-RFLP yielded 41 bands, 30 of which were polymorphic while ISSR produced 87 bands with 47 polymorphic bands. Both markers showed genetic variability among the analyzed isolates; however, IGS-RFLP was more efficient than ISSR, showing a higher polymorphic average (59.91%) than the latter (44.11%). The averages of polymorphic information content (PIC) were 0.211 and 0.129, respectively. Twenty haplotypes were identified by IGS-RFLP and 15 haplotypes by ISSR. Genotype clustering within dendrograms was different for both types of markers. The genetic groups obtained by IGS-RFLP showed a partial association to geographic origin. This is the first report on genetic variability of F. graminearum isolates from wheat in Argentina using IGS-RFLP and ISSR markers.

  9. Molecular evolution and population genetics of a Gram-negative binding protein gene in the malaria vector Anopheles gambiae (sensu lato)

    OpenAIRE

    Salgueiro, Patrícia; Lopes, Ana Sofia; Mendes, Cristina; Charlwood, Jacques Derek; Arez, Ana Paula; Pinto, João; Silveira, Henrique

    2016-01-01

    Background Clarifying the role of the innate immune system of the malaria vector Anopheles gambiae is a potential way to block the development of the Plasmodium parasites. Pathogen recognition is the first step of innate immune response, where pattern recognition proteins like GNBPs play a central role. Results We analysed 70 sequences of the protein coding gene GNBPB2 from two species, Anopheles gambiae (s.s.) and An. coluzzii, collected in six African countries. We detected 135 segregating ...

  10. The practical importance of permanent and semipermanent habitats for controlling aquatic stages of Anopheles gambiae sensu lato mosquitoes: operational observations from a rural town in western Kenya

    NARCIS (Netherlands)

    Fillinger, U.; Sonye, G.; Killeen, G.F.; Knols, B.G.J.; Becker, N.

    2004-01-01

    Control of aquatic-stage Anopheles is one of the oldest and most historically successful interventions to prevent malaria, but it has seen little application in Africa. Consequently, the ecology of immature afrotropical Anopheles has received insufficient attention. We therefore examined the populat

  11. Manipulating Steady Heat Conduction by Sensu-shaped Thermal Metamaterials

    CERN Document Server

    Han, Tiancheng; Liu, Dan; Gao, Dongliang; Li, Baowen; Thong, John T L; Qiu, Cheng-Wei

    2014-01-01

    The ability to design the control of heat flow has innumerable benefits in the design of electronic systems such as thermoelectric energy harvesters, solid-state lighting, and thermal imagers, where the thermal design plays a key role in performance and device reliability. However, to realize one advanced control function of thermal flux, one needs to design one sophisticated, multilayered and inhomogeneous thermal structure with different composition/shape at different regions of one device. In this work, we employ one identical sensu-unit with facile natural composition to experimentally realize a new class of thermal metamaterials for controlling thermal conduction (e.g., thermal concentrator, focusing/resolving, uniform heating), only resorting to positioning and locating the same unit element of sensu-shape structure. The thermal metamaterial unit and the proper arrangement of multiple identical units are capable of transferring, redistributing and managing thermal energy in a versatile fashion. It is al...

  12. Manipulating Steady Heat Conduction by Sensu-shaped Thermal Metamaterials

    Science.gov (United States)

    Han, Tiancheng; Bai, Xue; Liu, Dan; Gao, Dongliang; Li, Baowen; Thong, John T. L.; Qiu, Cheng-Wei

    2015-01-01

    The ability to design the control of heat flow has innumerable benefits in the design of electronic systems such as thermoelectric energy harvesters, solid-state lighting, and thermal imagers, where the thermal design plays a key role in performance and device reliability. In this work, we employ one identical sensu-unit with facile natural composition to experimentally realize a new class of thermal metamaterials for controlling thermal conduction (e.g., thermal concentrator, focusing/resolving, uniform heating), only resorting to positioning and locating the same unit element of sensu-shape structure. The thermal metamaterial unit and the proper arrangement of multiple identical units are capable of transferring, redistributing and managing thermal energy in a versatile fashion. It is also shown that our sensu-shape unit elements can be used in manipulating dc currents without any change in the layout for the thermal counterpart. These could markedly enhance the capabilities in thermal sensing, thermal imaging, thermal-energy storage, thermal packaging, thermal therapy, and more domains beyond. PMID:25974383

  13. Manipulating Steady Heat Conduction by Sensu-shaped Thermal Metamaterials.

    Science.gov (United States)

    Han, Tiancheng; Bai, Xue; Liu, Dan; Gao, Dongliang; Li, Baowen; Thong, John T L; Qiu, Cheng-Wei

    2015-05-14

    The ability to design the control of heat flow has innumerable benefits in the design of electronic systems such as thermoelectric energy harvesters, solid-state lighting, and thermal imagers, where the thermal design plays a key role in performance and device reliability. In this work, we employ one identical sensu-unit with facile natural composition to experimentally realize a new class of thermal metamaterials for controlling thermal conduction (e.g., thermal concentrator, focusing/resolving, uniform heating), only resorting to positioning and locating the same unit element of sensu-shape structure. The thermal metamaterial unit and the proper arrangement of multiple identical units are capable of transferring, redistributing and managing thermal energy in a versatile fashion. It is also shown that our sensu-shape unit elements can be used in manipulating dc currents without any change in the layout for the thermal counterpart. These could markedly enhance the capabilities in thermal sensing, thermal imaging, thermal-energy storage, thermal packaging, thermal therapy, and more domains beyond.

  14. 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.

  15. Socio-Economic Characteristics in Notified Erythema Migrans Patients

    Directory of Open Access Journals (Sweden)

    Sočan Maja

    2015-12-01

    Full Text Available Izhodišče. Lymska borelioza je posledica okužbe z bakterijami kompleksa Borrelia burgdorferi sensu lato. Erythema migrans (EM je najpogostejša klinična oblika lymske borelioze. Namen raziskave je bil pridobiti epidemiološke podatke in preučiti socialno-ekonomske značilnosti prijavljenih bolnikov z EM v Sloveniji.

  16. The search for true sources of TBEV and related pathogens : the danger of professed beliefs on pathogens, host and vector interplays

    OpenAIRE

    Gurung, Deepa

    2016-01-01

    Tick-borne disease in general is among the most important vector-borne diseases that are emerging as a threat to humans and is currently identified as a major health problem in many countries. Tick-borne encephalitis and lyme borreliosis are the main diseases transmitted by Ixodes ricinus ticks in Europe. Besides TBEV (Tick-borne encephalitis virus) and Borrelia burgdorferi sensu lato, I. ricinus is known to transmit other pathogenic microorganisms like- louping ill virus, Anaplasma phagocyto...

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

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    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.

  18. Lyme disease spirochaete Borrelia burgdorferi does not require thiamin.

    OpenAIRE

    Zhang, K; Bian, J; Deng, Y; Smith, A.; Nunez, RE; Li, MB; U. Pal; Yu, A-M; Qiu, W.; Ealick, SE; Li, C.

    2016-01-01

    Thiamin pyrophosphate (ThDP), the active form of thiamin (vitamin B1), is believed to be an essential cofactor for all living organisms1,2. Here, we report the unprecedented result that thiamin is dispensable for the growth of the Lyme disease pathogen Borrelia burgdorferi (Bb)3. Bb lacks genes for thiamin biosynthesis and transport as well as known ThDP-dependent enzymes4, and we were unable to detect thiamin or its derivatives in Bb cells. We showed that eliminating thiamin in vitro and in ...

  19. Lyme disease spirochaete Borrelia burgdorferi does not require thiamin

    OpenAIRE

    Zhang, Kai; Bian, Jiang; Deng, Yijie; Smith, Alexis; Nunez, Roy E.; Li, Michael B.; Pal, Utpal; Yu, Ai-Ming; Qiu, Weigang; Ealick, Steven E.; Li, Chunhao

    2016-01-01

    Thiamin pyrophosphate (ThDP), the active form of thiamin (vitamin B1), is believed to be an essential cofactor for all living organisms1,2. Here, we report the unprecedented result that thiamin is dispensable for the growth of the Lyme disease pathogen Borrelia burgdorferi (Bb)3. Bb lacks genes for thiamin biosynthesis and transport as well as known ThDP-dependent enzymes4, and we were unable to detect thiamin or its derivatives in Bb cells. We showed that eliminating thiamin in vitro and in ...

  20. Amplification of the flgE gene provides evidence for the existence of a Brazilian borreliosis

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    Elenice Mantovani

    2012-06-01

    Full Text Available INTRODUCTION: The symptoms of Brazilian borreliosis resemble the clinical manifestations of Lyme disease (LD. However, there are differences between the two in terms of epidemiological and laboratory findings. Primers usually employed to diagnose LD have failed to detect Borrelia strains in Brazil. OBJECTIVE: We aimed to identify the Brazilian Borrelia using a conserved gene that synthesizes the flagellar hook (flgE of Borrelia burgdorferi sensu lato. METHOD: Three patients presenting with erythema migrans and positive epidemiological histories were recruited for the study. Blood samples were collected, and the DNA was extracted by commercial kits. RESULTS: The gene flgE was amplified from DNA of all selected patients. Upon sequencing, these positive samples revealed 99% homology to B. burgdorferi flgE. CONCLUSION: These results support the existence of borreliosis in Brazil. However, it is unclear whether this borreliosis is caused by a genetically modified B. burgdorferi sensu stricto or by a new species of Borrelia spp.

  1. Season is an unreliable predictor of Lyme neuroborreliosis

    DEFF Research Database (Denmark)

    Petersen, Bo Bødker; Møller, Jens Kjølseth; Vilholm, Ole Jakob

    2015-01-01

    INTRODUCTION: Lyme neuroborreliosis (LNB) is a tick-borne infection of the nervous system caused by the spirochete Borrelia burgdorferi sensu lato. The primary symptoms are usually painful radiculitis, facial palsy and lymphocytic meningitis. The aim of this study was to provide data...... of leucocytosis in the cerebrospinal fluid and intrathecal production of immunoglobulin M and/or G anti-B. burgdorferi antibodies. RESULTS: Onset of neurological symptoms in LNB occurred year round in the Region of Southern Denmark. Only half of the patients had a history of a tick bite or erythema migrans (EM...

  2. Infezione da Borrelia burgdorferi: descrizione di un caso clinico atipico

    Directory of Open Access Journals (Sweden)

    Gino Ciarrocchi

    2005-06-01

    Full Text Available La borreliosi o Malattia di Lyme (ML è una malattia infettiva multisistemica causata da una spirocheta, Borrelia burgdorferi, trasmessa dal morso di una zecca del genere Ixodes.Viene descritto il caso clinico di una ragazza di venti anni. In un soggiorno estivo sul Monte Amiata, in Toscana, la giovane notò un arrossamento della pelle al fianco sinistro del corpo. Non osservò presenza della zecca nella zona di presumibile inoculo, da cui si sviluppò un eritema migrante (EM ad andamento centrifugo, fino a raggiungere un diametro di circa 10 centimetri. Nei giorni successivi e durante il follow up non sono mai comparsi sintomi collaterali generali riferibili ad astenia, febbricola, artralgie, malessere, né a carico di organi bersaglio. Dopo circa due settimane, persistendo l’EM, sono stati eseguiti esami sierologici specifici per Borrelia burgdorferi. Al riscontro positivo dei test ELISA e western-blot IgG e IgM, il clnico ha prescritto terapia antibiotica a base di Amoxicillina 2g/die per 20 giorni.Dopo 8-10 giorni di terapia è avvenuta la scomparsa completa dell’eritema. Viene mostrato l’andamento dei test sierologici al momento della diagnosi e durante il follow up, per un periodo di circa cinque mesi.

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

    DEFF Research Database (Denmark)

    Lebech, Anne-Mette

    2002-01-01

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

  4. 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.

  5. 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.

  6. A catalog of the types of Chrysomelidae sensu lato (Insecta, Coleoptera, Polyphaga deposited in the Museo Argentino de Ciencias Naturales, Buenos Aires Catálogo de los tipos de Chrysomelidae sensu lato (Insecta, Coleoptera, Polyphaga depositados en el Museo Argentino de Ciencias Naturales, Buenos Aires

    Directory of Open Access Journals (Sweden)

    Axel O. Bachmann

    2010-06-01

    Full Text Available The type specimens (all current categories of Chrysomelidae s.l. deposited in this Museum are listed; 125 names are recorded, 85 of them (68 percent are represented here by name-bearing types ('primary' types, five of them dubious. The family is taken in its broadest sense, including the Bruchinae, Hispinae (along with the former Cassidinae, and other groups sometimes considered as separate families. The specific and subspecific names were alphabetically filed, followed by the generic ones as they were spelled in the original publication, or the generic and specific names in the case of subspecies and varieties. Later combinations and/or current binomina are mentioned insofar as these are known to the authors. Two lists are added: 1. of specimens labelled as types of unavailable names, chiefly those not found in the literature, and supposedly not published, and 2. of specimens labelled as types, but not originally included as such, and published or not after the original description.Se catalogan los ejemplares típicos, de todas las categorías aceptadas, de Chrysomelidae s.l. conservados en este museo; se registran 125 nombres, 85 de ellos (68 por ciento representados aquí por tipos portadores de nombres (tipos 'primarios', cinco de ellos dudosos. La familia se toma en su concepto más amplio, incluyendo a las Bruchinae, las Hispinae (con las anteriores Cassidinae y otros grupos a veces considerados como familias separadas. Los nombres específicos y subespecíficos fueron ordenados alfabéticamente; a estos siguen los de los géneros, así como se publicaron originalmente, o de los géneros y especies en el caso de las subespecies y variedades. Se mencionan las combinaciones ulteriores y/o los binomios en uso, hasta donde son conocidos por los autores. Se agregan dos listas: 1. de los ejemplares rotulados como tipos, de nombres no disponibles, principalmente aquellos no hallados en la bibliografía, y supuestamente no publicados, y 2. de los ejemplares rotulados como tipos, pero no incluidos originalmente como tales, publicados o no después de la descripción original.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. 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.

  9. 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.

  10. 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

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

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    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

  12. 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.

  13. A Manganese-rich Environment Supports Superoxide Dismutase Activity in a Lyme Disease Pathogen, Borrelia burgdorferi*

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    Aguirre, J. Dafhne; Clark, Hillary M.; McIlvin, Matthew; Vazquez, Christine; Palmere, Shaina L.; Grab, Dennis J.; Seshu, J.; Hart, P. John; Saito, Mak; Culotta, Valeria C.

    2013-01-01

    The Lyme disease pathogen Borrelia burgdorferi represents a novel organism in which to study metalloprotein biology in that this spirochete has uniquely evolved with no requirement for iron. Not only is iron low, but we show here that B. burgdorferi has the capacity to accumulate remarkably high levels of manganese. This high manganese is necessary to activate the SodA superoxide dismutase (SOD) essential for virulence. Using a metalloproteomic approach, we demonstrate that a bulk of B. burgdorferi SodA directly associates with manganese, and a smaller pool of inactive enzyme accumulates as apoprotein. Other metalloproteins may have similarly adapted to using manganese as co-factor, including the BB0366 aminopeptidase. Whereas B. burgdorferi SodA has evolved in a manganese-rich, iron-poor environment, the opposite is true for Mn-SODs of organisms such as Escherichia coli and bakers' yeast. These Mn-SODs still capture manganese in an iron-rich cell, and we tested whether the same is true for Borrelia SodA. When expressed in the iron-rich mitochondria of Saccharomyces cerevisiae, B. burgdorferi SodA was inactive. Activity was only possible when cells accumulated extremely high levels of manganese that exceeded cellular iron. Moreover, there was no evidence for iron inactivation of the SOD. B. burgdorferi SodA shows strong overall homology with other members of the Mn-SOD family, but computer-assisted modeling revealed some unusual features of the hydrogen bonding network near the enzyme's active site. The unique properties of B. burgdorferi SodA may represent adaptation to expression in the manganese-rich and iron-poor environment of the spirochete. PMID:23376276

  14. A manganese-rich environment supports superoxide dismutase activity in a Lyme disease pathogen, Borrelia burgdorferi.

    Science.gov (United States)

    Aguirre, J Dafhne; Clark, Hillary M; McIlvin, Matthew; Vazquez, Christine; Palmere, Shaina L; Grab, Dennis J; Seshu, J; Hart, P John; Saito, Mak; Culotta, Valeria C

    2013-03-22

    The Lyme disease pathogen Borrelia burgdorferi represents a novel organism in which to study metalloprotein biology in that this spirochete has uniquely evolved with no requirement for iron. Not only is iron low, but we show here that B. burgdorferi has the capacity to accumulate remarkably high levels of manganese. This high manganese is necessary to activate the SodA superoxide dismutase (SOD) essential for virulence. Using a metalloproteomic approach, we demonstrate that a bulk of B. burgdorferi SodA directly associates with manganese, and a smaller pool of inactive enzyme accumulates as apoprotein. Other metalloproteins may have similarly adapted to using manganese as co-factor, including the BB0366 aminopeptidase. Whereas B. burgdorferi SodA has evolved in a manganese-rich, iron-poor environment, the opposite is true for Mn-SODs of organisms such as Escherichia coli and bakers' yeast. These Mn-SODs still capture manganese in an iron-rich cell, and we tested whether the same is true for Borrelia SodA. When expressed in the iron-rich mitochondria of Saccharomyces cerevisiae, B. burgdorferi SodA was inactive. Activity was only possible when cells accumulated extremely high levels of manganese that exceeded cellular iron. Moreover, there was no evidence for iron inactivation of the SOD. B. burgdorferi SodA shows strong overall homology with other members of the Mn-SOD family, but computer-assisted modeling revealed some unusual features of the hydrogen bonding network near the enzyme's active site. The unique properties of B. burgdorferi SodA may represent adaptation to expression in the manganese-rich and iron-poor environment of the spirochete.

  15. The occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs in China.

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    Xia, Z; Yu, D; Mao, J; Zhang, Z; Yu, J

    2012-06-01

    A survey of the occurrence of Dirofilaria immitis, Borrelia burgdorferi, Ehrlichia canis and Anaplasma phagocytophium in dogs was undertaken in the People's Republic of China between October 2008 and October 2009. A total of 600 blood samples were taken from dogs in four cities in China: 300 in Beijing, 150 in Shenzhen, 30 in Shanghai and 120 in Zhengzhou. All samples were tested for the heartworm antigen and antibodies of canine B. burgdorferi, E. canis and A. phagocytophium by using the canine SNAP® 4Dx® test kit. The occurrence of D. immitis, B. burgdorferi, E. canis and A. phagocytophium was 1.17% (7/600), 0.17% (1/600), 2.17% (13/600) and 0.5% (3/600), respectively. In Shenzhen city 2% (3/150), 8.67% (13/150) and 2% (3/150) of samples were positive for D. immitis, E. canis and A. phagocytophium, respectively. The occurrence of heartworm antigen was 0.33% (1/300) in Beijing, 2.00% (3/150) in Shenzhen, 3.33% (1/30) in Shanghai and 1.67% (2/120) in Zhengzhou. We found E. canis and A. phagocytophium only at one site, Shenzhen, while the only occurrence of B. burgdorferi was at Beijing. In conclusion, the dog population in China is at potential risk for D. immitis, B. burgdorferi, E. canis and A. phagocytophium infection, the risk being especially high in southern China.

  16. 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.

  17. Persistence of Borrelia burgdorferi in rhesus macaques following antibiotic treatment of disseminated infection.

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    Monica E Embers

    Full Text Available The persistence of symptoms in Lyme disease patients following antibiotic therapy, and their causes, continue to be a matter of intense controversy. The studies presented here explore antibiotic efficacy using nonhuman primates. Rhesus macaques were infected with B. burgdorferi and a portion received aggressive antibiotic therapy 4-6 months later. Multiple methods were utilized for detection of residual organisms, including the feeding of lab-reared ticks on monkeys (xenodiagnosis, culture, immunofluorescence and PCR. Antibody responses to the B. burgdorferi-specific C6 diagnostic peptide were measured longitudinally and declined in all treated animals. B. burgdorferi antigen, DNA and RNA were detected in the tissues of treated animals. Finally, small numbers of intact spirochetes were recovered by xenodiagnosis from treated monkeys. These results demonstrate that B. burgdorferi can withstand antibiotic treatment, administered post-dissemination, in a primate host. Though B. burgdorferi is not known to possess resistance mechanisms and is susceptible to the standard antibiotics (doxycycline, ceftriaxone in vitro, it appears to become tolerant post-dissemination in the primate host. This finding raises important questions about the pathogenicity of antibiotic-tolerant persisters and whether or not they can contribute to symptoms post-treatment.

  18. 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.

  19. [Pedagogical training in stricto sensu graduate programs in public health].

    Science.gov (United States)

    Corrêa, Guilherme Torres; Ribeiro, Victoria Maria Brant

    2013-06-01

    The scope of this research is to discuss the relevance and need for pedagogical training of university lecturers in the Public Health field. The contention is that college teaching is a practice that requires specific training, since it is characterized by complex elements that transcend the mastery of given content. Considering stricto sensu graduate studies as an important stage in the training of future university lecturers, an attempt was made to identify and analyze the subjects and practices of pedagogical training in academic masters and doctorate programs in Public Health. To achieve the research aim, this work was based on Pierre Bourdieu's field theory and on Tomaz Tadeu da Silva's curriculum theory. Results indicate that the programs do not consider the aspect of teacher training as a major issue. With regard to the Public Health field approximately 61% of masters and 38% of doctorate programs have pedagogical training subjects/practices. Furthermore, there is a tendency for technical-instrumental training, which is in line with the history of the Public Health field. The conclusion is that there is a need to develop a culture that values college and graduate Public Health teaching, considering the complexity of pedagogical practice in all its dimensions.

  20. Lyme disease spirochaete Borrelia burgdorferi does not require thiamin.

    Science.gov (United States)

    Zhang, Kai; Bian, Jiang; Deng, Yijie; Smith, Alexis; Nunez, Roy E; Li, Michael B; Pal, Utpal; Yu, Ai-Ming; Qiu, Weigang; Ealick, Steven E; Li, Chunhao

    2016-11-21

    Thiamin pyrophosphate (ThDP), the active form of thiamin (vitamin B1), is believed to be an essential cofactor for all living organisms(1,2). Here, we report the unprecedented result that thiamin is dispensable for the growth of the Lyme disease pathogen Borrelia burgdorferi (Bb)(3). Bb lacks genes for thiamin biosynthesis and transport as well as known ThDP-dependent enzymes(4), and we were unable to detect thiamin or its derivatives in Bb cells. We showed that eliminating thiamin in vitro and in vivo using BcmE, an enzyme that degrades thiamin, has no impact on Bb growth and survival during its enzootic infectious cycle. Finally, high-performance liquid chromatography analysis reveals that the level of thiamin and its derivatives in Ixodes scapularis ticks, the enzootic vector of Bb, is extremely low. These results suggest that by dispensing with use of thiamin, Borrelia, and perhaps other tick-transmitted bacterial pathogens, are uniquely adapted to survive in tick vectors before transmitting to mammalian hosts. To our knowledge, such a mechanism has not been reported previously in any living organisms.

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

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    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.

  2. 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

  3. Borrelia burgdorferi Keeps Moving and Carries on: A Review of Borrelial Dissemination and Invasion

    Science.gov (United States)

    Hyde, Jenny A.

    2017-01-01

    Borrelia burgdorferi is the etiological agent of Lyme disease, a multisystemic, multistage, inflammatory infection resulting in patients experiencing cardiac, neurological, and arthritic complications when not treated with antibiotics shortly after exposure. The spirochetal bacterium transmits through the Ixodes vector colonizing the dermis of a mammalian host prior to hematogenous dissemination and invasion of distal tissues all the while combating the immune response as it traverses through its pathogenic lifecycle. The innate immune response controls the borrelial burden in the dermis, but is unable to clear the infection and thereby prevent progression of disease. Dissemination in the mammalian host requires temporal regulation of virulence determinants to allow for vascular interactions, invasion, and colonization of distal tissues. Virulence determinants and/or adhesins are highly heterogenetic among environmental B. burgdorferi strains with particular genotypes being associated with the ability to disseminate to specific tissues and the severity of disease, but fail to generate cross-protective immunity between borrelial strains. The unique motility of B. burgdorferi rendered by the endoflagella serves a vital function for dissemination and protection from immune recognition. Progress has been made toward understanding the chemotactic regulation coordinating the activity of the two polar localized flagellar motors and their role in borrelial virulence, but this regulation is not yet fully understood. Distinct states of motility allow for dynamic interactions between several B. burgdorferi adhesins and host targets that play roles in transendothelial migration. Transmigration across endothelial and blood–brain barriers allows for the invasion of tissues and elicits localized immune responses. The invasive nature of B. burgdorferi is lacking in proactive mechanisms to modulate disease, such as secretion systems and toxins, but recent work has shown

  4. Prevalence of Borrelia burgdorferi-infected ticks from wildlife hosts, a response to Norris et al.

    Science.gov (United States)

    Esteve-Gassent, Maria D; Grover, Abha; Feria-Arroyo, Teresa P; Castro-Arellano, Ivan; Medina, Raul F; Gordillo-Pérez, Guadalupe; de León, Adalberto A Pérez

    2015-02-27

    In a recent Letter to the Editor, Norris et al. questioned the validity of some of our data reported by Feria-Arroyo et al. The main issue investigated by us was the potential impact of climate change on the probable distribution of the tick vector Ixodes scapularis in the Texas-Mexico transboundary region. As an ancillary issue, an analysis of sequence data for the intergenic spacer of Borrelia burgdorferi was conducted. In the present letter, we provide further evidence supporting our original results, and advocate that extensive study of the population genetics of B. burgdorferi is needed in the Texas-Mexico transboundary region.

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

    Directory of Open Access Journals (Sweden)

    Angela de Oliveira

    2004-06-01

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

  6. A novel zoonotic genotype related to Echinococcus granulosus sensu stricto from southern Ethiopia.

    Science.gov (United States)

    Wassermann, Marion; Woldeyes, Daniel; Gerbi, Banchwosen Mechal; Ebi, Dennis; Zeyhle, Eberhard; Mackenstedt, Ute; Petros, Beyene; Tilahun, Getachew; Kern, Peter; Romig, Thomas

    2016-09-01

    Complete mitochondrial and two nuclear gene sequences of a novel genotype (GOmo) related to Echinococcus granulosus sensu stricto are described from a metacestode isolate retrieved from a human patient in southwestern Ethiopia. Phylogenetically, the genotype is positioned within the E. granulosus sensu stricto/Echinococcus felidis cluster, but cannot easily be allocated to either species. Based on different mitochondrial DNA markers, it is closest to the haplotype cluster that currently defines the species E. granulosus sensu stricto (which includes variants showing the widely cited G1, G2 and G3 sequences), but is clearly not part of this cluster. Pairwise distances between GOmo and E. granulosus sensu stricto are in the range of those between the most distant members of the Echinococcus canadensis complex (G6-10) that were recently proposed as separate species. At this stage, we prefer to list GOmo informally as a genotype rather than giving it any taxonomic rank because our knowledge rests on a single isolate from a dead-end host (human), and its lifecycle is unknown. According to data on molecularly characterised Echinococcus isolates from this region, GOmo has never been found in the usual livestock species that carry cystic echinococcosis and the possibility of a wildlife source of this newly recognised zoonotic agent cannot be excluded. The discovery of GOmo adds complexity to the already diverse array of cystic echinococcosis agents in sub-Saharan Africa and challenges hypotheses on the biogeographical origin of the E. granulosus sensu stricto clade.

  7. Coexistence of emerging bacterial pathogens in Ixodes ricinus ticks in Serbia*

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    Tomanović S.

    2010-09-01

    Full Text Available The list of tick-borne pathogens is long, varied and includes viruses, bacteria, protozoa and nematodes. As all of these agents can exist in ticks, their co-infections have been previously reported. We studied co-infections of emerging bacterial pathogens (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Francisella tularensis in Ixodes ricinus ticks in Serbia. Using PCR technique, we detected species-specific sequences, rrf-rrl rDNA intergenic spacer for B. burgdorferi s.l., p44/msp2 paralogs for A. phagocytophilum, and the 17 kDa lipoprotein gene, TUL4, for F. tularensis, respectively, in total DNA extracted from the ticks. Common infections with more than one pathogen were detected in 42 (28.8 % of 146 infected I. ricinus ticks. Co-infections with two pathogens were present in 39 (26.7 % of infected ticks. Simultaneous presence of A. phagocytophilum and different genospecies of B. burgdorferi s.l. complex was recorded in 16 ticks, co-infection with different B. burgdorferi s. l. genospecies was found in 15 ticks and eight ticks harbored mixed infections with F. tularensis and B. burgdorferi s.l. genospecies. Less common were triple pathogen species infections, detected in three ticks, one infected with A. phagocytophilum / B. burgdorferi s.s. / B. lusitaniae and two infected with F. tularensis / B. burgdorferi s.s. / B. lusitaniae. No mixed infections of A. phagocytophilum and F. tularensis were detected.

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

    NARCIS (Netherlands)

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

    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

  9. Human Coinfection with Borrelia burgdorferi and Babesia microti in the United States

    Directory of Open Access Journals (Sweden)

    Kristen L. Knapp

    2015-01-01

    Full Text Available Borrelia burgdorferi, the causative agent of Lyme disease, and Babesia microti, a causative agent of babesiosis, are increasingly implicated in the growing tick-borne disease burden in the northeastern United States. These pathogens are transmitted via the bite of an infected tick vector, Ixodes scapularis, which is capable of harboring and inoculating a host with multiple pathogens simultaneously. Clinical presentation of the diseases is heterogeneous and ranges from mild flu-like symptoms to near-fatal cardiac arrhythmias. While the reason for the variability is not known, the possibility exists that concomitant infection with both B. burgdorferi and B. microti may synergistically increase disease severity. In an effort to clarify the current state of understanding regarding coinfection with B. burgdorferi and B. microti, in this review, we discuss the geographical distribution and pathogenesis of Lyme disease and babesiosis in the United States, the immunological response of humans to B. burgdorferi or B. microti infection, the existing knowledge regarding coinfection disease pathology, and critical factors that have led to ambiguity in the literature regarding coinfection, in order to eliminate confusion in future experimental design and investigation.

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

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    Tanja Petnicki-Ocwieja

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

  11. [Serological diagnosis of sporotrichosis using an antigen of Sporothrix schenckii sensu stricto mycelium].

    Science.gov (United States)

    Alvarado, Primavera; Ostos, Ana; Franquiz, Nohelys; Roschman-González, Antonio; Zambrano, Edgar A; Mendoza, Mireya

    2015-06-01

    We developed and analyzed an Enzyme-Linked Immunosorbent Assay (ELISA) in order to detect antibodies in sera from sporotrichosis patients. We used a crude antigen of Sporothrix schenckii sensu stricto, obtained from the mycelial phase of the fungi. Positive sera were analyzed by other serological techniques such as double immunodiffusion (IGG) and counterimmunoelectrophoresis (CIE). The assay was validated by using sera from patients with other pathologies such as: histoplasmosis, paracoccidioidomycosis, tuberculosis, leishmaniasis, lupus and healthy individuals as negative controls. For the Sporothrix schenckii sensu stricto antigen, we found a 100% of specificity by every technique and sensitivity higher than 98% with IDD, CIE and ELISA. Our results show a high sensitivity and specificity for the Sporothrix schenckii sensu stricto antigen, so it can be used for IDD, CIE and ELISA. The results suggest that this antigen could be used in conjunction with other conventional tests for differential diagnosis and may be useful for monitoring the disease progression and response to treatment.

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

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    John J. Lazarus

    2012-01-01

    Full Text Available 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-specific IgM and IgG than groups receiving large inocula of heat-killed bacteria. Additionally, sera from mice injected with varied doses of killed B. burgdorferi recognized unique borrelial antigens compared to mice infected with live B. burgdorferi. Intradermal injection of killed B. burgdorferi resulted in rapid DNA clearance from skin, whereas DNA was consistently detected in skin inoculated with viable B. burgdorferi. These data indicate that both ELISA-based serological analyses and PCR-based methods of assessing B. burgdorferi infection clearly distinguish between an established infection with live bacteria and exposure to large numbers of bacteria that are promptly cleared by the innate responses.

  13. 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.

  14. 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.

  15. Borrelia burgdorferi promotes the establishment of Babesia microti in the northeastern United States.

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    Jessica M Dunn

    Full Text Available Babesia microti and Borrelia burgdorferi, the respective causative agents of human babesiosis and Lyme disease, are maintained in their enzootic cycles by the blacklegged tick (Ixodes scapularis and use the white-footed mouse (Peromyscus leucopus as primary reservoir host. The geographic range of both pathogens has expanded in the United States, but the spread of babesiosis has lagged behind that of Lyme disease. Several studies have estimated the basic reproduction number (R0 for B. microti to be below the threshold for persistence (<1, a finding that is inconsistent with the persistence and geographic expansion of this pathogen. We tested the hypothesis that host coinfection with B. burgdorferi increases the likelihood of B. microti transmission and establishment in new areas. We fed I. scapularis larva on P. leucopus mice that had been infected in the laboratory with B. microti and/or B. burgdorferi. We observed that coinfection in mice increases the frequency of B. microti infected ticks. To identify the ecological variables that would increase the probability of B. microti establishment in the field, we integrated our laboratory data with field data on tick burden and feeding activity in an R0 model. Our model predicts that high prevalence of B. burgdorferi infected mice lowers the ecological threshold for B. microti establishment, especially at sites where larval burden on P. leucopus is lower and where larvae feed simultaneously or soon after nymphs infect mice, when most of the transmission enhancement due to coinfection occurs. Our studies suggest that B. burgdorferi contributes to the emergence and expansion of B. microti and provides a model to predict the ecological factors that are sufficient for emergence of B. microti in the wild.

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

    Science.gov (United States)

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

    2008-12-16

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

  17. Identification of Borrelia species after creation of an in-house MALDI-TOF MS database.

    Directory of Open Access Journals (Sweden)

    Adriana Calderaro

    Full Text Available Lyme borreliosis (LB is a multisystemic disease caused by Borrelia burgdorferi sensu lato (sl complex transmitted to humans by Ixodes ticks. B. burgdorferi sl complex, currently comprising at least 19 genospecies, includes the main pathogenic species responsible for human disease in Europe: B. burgdorferi sensu stricto (ss, B. afzelii, and B. garinii. In this study, for the first time, MALDI-TOF MS was applied to Borrelia spp., supplementing the existing database, limited to the species B. burgdorferi ss, B . spielmanii and B. garinii, with the species B. afzelii, in order to enable the identification of all the species potentially implicated in LB in Europe. Moreover, we supplemented the database also with B. hermsii, which is the primary cause of tick-borne relapsing fever in western North America, B. japonica, circulating in Asia, and another reference strain of B. burgdorferi ss (B31 strain. The dendrogram obtained by analyzing the protein profiles of the different Borrelia species reflected Borrelia taxonomy, showing that all the species included in the Borrelia sl complex clustered in a unique branch, while Borrelia hermsii clustered separately. In conclusion, in this study MALDI-TOF MS proved a useful tool suitable for identification of Borrelia spp. both for diagnostic purpose and epidemiological surveillance.

  18. Strictu sensu post-graduation in anesthesiology: experience of ten years at Universidade Estadual Paulista

    OpenAIRE

    José Reinaldo Cerqueira Braz; Pedro Thadeu Galvão Vianna; Yara Marcondes Machado Castiglia; Luiz Antonio Vane; Flávio Massone; Lino Lemonica; Gladys Bastos de Castro

    2005-01-01

    JUSTIFICATIVA E OBJETIVOS: O número de Programas de Pós-Graduação em Anestesiologia stricto sensu existente no país ainda é muito pequeno. Com a finalidade de incentivar a pós-graduação em Anestesiologia no Brasil, é apresentada a experiência acumulada em dez anos de atividades do programa da Universidade Estadual Paulista (UNESP). CONTEÚDO: O Programa de Pós-Graduação em Anestesiologia stricto sensu da UNESP foi credenciado pela CAPES, desde a sua criação em 1994, nos Cursos de Mestrado e Do...

  19. 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.

  20. 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.

  1. An optimized SYBR Green I/PI assay for rapid viability assessment and antibiotic susceptibility testing for Borrelia burgdorferi.

    Science.gov (United States)

    Feng, Jie; Wang, Ting; Zhang, Shuo; Shi, Wanliang; Zhang, Ying

    2014-01-01

    Lyme disease caused by Borrelia burgdorferi is the most common tick-borne disease in the US and Europe. Unlike most bacteria, measurements of growth and viability of B. burgdorferi are challenging. The current B. burgdorferi viability assays based on microscopic counting and PCR are cumbersome and tedious and cannot be used in a high throughput format. Here, we evaluated several commonly used viability assays including MTT and XTT assays, fluorescein diacetate assay, Sytox Green/Hoechst 33342 assay, the commercially available LIVE/DEAD BacLight assay, and SYBR Green I/PI assay by microscopic counting and by automated 96-well plate reader for rapid viability assessment of B. burgdorferi. We found that the optimized SYBR Green I/PI assay based on green to red fluorescence ratio is superior to all the other assays for measuring the viability of B. burgdorferi in terms of sensitivity, accuracy, reliability, and speed in automated 96-well plate format and in comparison with microscopic counting. The BSK-H medium which produced a high background for the LIVE/DEAD BacLight assay did not affect the SYBR Green I/PI assay, and the viability of B. burgdorferi culture could be directly measured using a microtiter plate reader. The SYBR Green I/PI assay was found to reliably assess the viability of planktonic as well as biofilm B. burgdorferi and could be used as a rapid antibiotic susceptibility test. Thus, the SYBR Green I/PI assay provides a more sensitive, rapid and convenient method for evaluating viability and antibiotic susceptibility of B. burgdorferi and can be used for high-throughput drug screens.

  2. Borrelia burgdorferi, a Pathogen That Lacks Iron, Encodes Manganese-dependent Superoxide Dismutase Essential for Resistance to Streptonigrin*

    Science.gov (United States)

    Troxell, Bryan; Xu, Haijun; Yang, X. Frank

    2012-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease, exists in nature through a complex life cycle involving ticks of the Ixodes genus and mammalian hosts. During its life cycle, B. burgdorferi experiences fluctuations in oxygen tension and may encounter reactive oxygen species (ROS). The key metalloenzyme to degrade ROS in B. burgdorferi is SodA. Although previous work suggests that B. burgdorferi SodA is an iron-dependent superoxide dismutase (SOD), later work demonstrates that B. burgdorferi is unable to transport iron and contains an extremely low intracellular concentration of iron. Consequently, the metal cofactor for SodA has been postulated to be manganese. However, experimental evidence to support this hypothesis remains lacking. In this study, we provide biochemical and genetic data showing that SodA is a manganese-dependent enzyme. First, B. burgdorferi contained SOD activity that is resistant to H2O2 and NaCN, characteristics associated with Mn-SODs. Second, the addition of manganese to the Chelex-treated BSK-II enhanced SodA expression. Third, disruption of the manganese transporter gene bmtA, which significantly lowers the intracellular manganese, greatly reduced SOD activity and SodA expression, suggesting that manganese regulates the level of SodA. In addition, we show that B. burgdorferi is resistant to streptonigrin, a metal-dependent redox cycling compound that produces ROS, and that SodA plays a protective role against the streptonigrin. Taken together, our data demonstrate the Lyme disease spirochete encodes a manganese-dependent SOD that contributes to B. burgdorferi defense against intracellular superoxide. PMID:22500025

  3. Borrelia burgdorferi, a pathogen that lacks iron, encodes manganese-dependent superoxide dismutase essential for resistance to streptonigrin.

    Science.gov (United States)

    Troxell, Bryan; Xu, Haijun; Yang, X Frank

    2012-06-01

    Borrelia burgdorferi, the causative agent of Lyme disease, exists in nature through a complex life cycle involving ticks of the Ixodes genus and mammalian hosts. During its life cycle, B. burgdorferi experiences fluctuations in oxygen tension and may encounter reactive oxygen species (ROS). The key metalloenzyme to degrade ROS in B. burgdorferi is SodA. Although previous work suggests that B. burgdorferi SodA is an iron-dependent superoxide dismutase (SOD), later work demonstrates that B. burgdorferi is unable to transport iron and contains an extremely low intracellular concentration of iron. Consequently, the metal cofactor for SodA has been postulated to be manganese. However, experimental evidence to support this hypothesis remains lacking. In this study, we provide biochemical and genetic data showing that SodA is a manganese-dependent enzyme. First, B. burgdorferi contained SOD activity that is resistant to H(2)O(2) and NaCN, characteristics associated with Mn-SODs. Second, the addition of manganese to the Chelex-treated BSK-II enhanced SodA expression. Third, disruption of the manganese transporter gene bmtA, which significantly lowers the intracellular manganese, greatly reduced SOD activity and SodA expression, suggesting that manganese regulates the level of SodA. In addition, we show that B. burgdorferi is resistant to streptonigrin, a metal-dependent redox cycling compound that produces ROS, and that SodA plays a protective role against the streptonigrin. Taken together, our data demonstrate the Lyme disease spirochete encodes a manganese-dependent SOD that contributes to B. burgdorferi defense against intracellular superoxide.

  4. 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

    By crossed immunoelectrophoresis and Western blotting (immunoblotting), it was shown that Borrelia burgdorferi expresses the 60-kilodalton Common Antigen (CA) that is cross-reactive with an equivalent antigen in a wide range of remotely related bacteria. B. burgdorferi CA is strongly immunogenic....... A B. burgdorferi genomic library was constructed by using a plasmid cloning system. Escherichia coli recombinants were screened for expression of immunodominant B. burgdorferi antigens. One of the recombinant clones expressed the 60-kilodalton CA of B. burgdorferi. The DNA region encoding B....... 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...

  5. Studies on Barbula consanguinea (Thw. & Mitt.) Jaeg. sensu Eddy : a pan-tropical species

    OpenAIRE

    Sollman, Philip

    2000-01-01

    Barbula consanguinea (Thw. & Mitt.) Jaeg. sensu Eddy is considered as a pan-tropical species. This taxon is reported new for several central African countries, the Arabian peninsula, and Middle America. This variable species is shortly discussed and compared with (most) related taxa. Hymenostylium crispulum Broth. & Par. and Barbula obscura Sull. (= Barbula wrightii Sauerb.) are considered as synonyms of Barbula consanguinea.

  6. Effectiveness of synthetic versus natural human volatiles as attractants for Anopheles gambiae (Diptera: Culicidae) sensu stricto

    NARCIS (Netherlands)

    Smallegange, R.C.; Knols, B.G.J.; Takken, W.

    2010-01-01

    Females of the African malaria vector, Anopheles gambiae Giles sensu stricto, use human volatiles to find their blood-host. Previous work has shown that ammonia, lactic acid, and aliphatic carboxylic acids significantly affect host orientation and attraction of this species, In the current study, th

  7. 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

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

    DEFF Research Database (Denmark)

    Lebech, Anne-Mette

    2002-01-01

    Lyme borreliosis caused by the spirochete Borrelia burgdorferi is now the most common vectorborne disease in North America, Europe and Asia. It is a multisystemic infection which may cause skin, neurological, cardiac or rheumatologic disorders. The aims of the present thesis were: (i) to develop...... for PCR amplification and subsequent identification of B. burgdorferi specific sequences were established and used. For all assays the analytical sensitivity was a few genome copies using purified DNA as template. The efficacy of PCR was initially evaluated using tissue samples from experimentally...... was detectable in 17-21% of CSF samples from patients with neuroborreliosis. In patients with very early neuroborreliosis (disease duration. PCR can be used as a diagnostic...

  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. 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

    Jutland and Funen, while 11% were positive for Borrelia burgdorferi. The Borrelia genotype B. afzelii was most prevalent, followed by B. valaisiana, B. burgdorferi s.s. and B. garinii.A. phagocytophilum was found in 14.5% of nymphs and 40.5% of adult ticks, while Borrelia was found in 13% of nymphs and 8......Borreliosis is an endemic infection in Denmark. Recent serosurveys have indicated that human anaplasmosis may be equally common. The aim of this study was to look for Anaplasma phagocytophilum and related pathogens in Ixodes ricinus ticks and estimate their prevalence, compared to Borrelia, using......% 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...

  11. 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.

  12. 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.

  13. 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

  14. Human Coinfection with Borrelia burgdorferi and Babesia microti in the United States

    OpenAIRE

    2015-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease, and Babesia microti, a causative agent of babesiosis, are increasingly implicated in the growing tick-borne disease burden in the northeastern United States. These pathogens are transmitted via the bite of an infected tick vector, Ixodes scapularis, which is capable of harboring and inoculating a host with multiple pathogens simultaneously. Clinical presentation of the diseases is heterogeneous and ranges from mild flu-like symptoms t...

  15. Altered Murine Tissue Colonization by Borrelia burgdorferi following Targeted Deletion of Linear Plasmid 17-Carried Genes

    OpenAIRE

    Casselli, Timothy; Tourand, Yvonne; Bankhead, Troy

    2012-01-01

    The causative agent of Lyme disease, Borrelia burgdorferi, possesses a segmented genome comprised of a single linear chromosome and upwards of 23 linear and circular plasmids. Much of what is known about plasmid-borne genes comes from studying laboratory clones that have spontaneously lost one or more plasmids during in vitro passage. Some plasmids, including the linear plasmid lp17, are never or rarely reported to be lost during routine culture; therefore, little is known about the requireme...

  16. 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.

  17. 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.

  18. 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.

  19. Ultrastructure of Spironucleus salmonis n. comb. (formerly Octomitus salmonis sensu Moore 1922, Davis 1926, and Hexamita salmonis sensu Ferguson 1979), with a guide to spironucleus species.

    Science.gov (United States)

    Poynton, Sarah L; Fard, M Reza Saghari; Jenkins, John; Ferguson, Hugh W

    2004-07-05

    Diplomonad flagellates can be associated with significant morbidity and mortality in fishes, particularly in farmed salmonids. Diagnosis using only light microscopy is limited, and species cannot be confirmed. We therefore undertook a comprehensive transmission electron microscopy study of 20 trophozoites from the intestine of farmed juvenile rainbow trout Oncorhynchus mykiss from Northern Ireland (reported as Hexamita salmonis by Ferguson in 1979). Re-assignment to the genus Spironucleus was determined based on the anteriorly tapering and intertwined elongate nuclei, anterior-medial kinetosomes, and the presence of a flagellar pocket. At the species level we observed a tri-radiate pattern of microtubules in the opening of the asymetrical striated lamina, comprising 3 to 5 microtubules following the edge of the flagellar pocket, 3 radiating away from the opening of the striated lamina, and 4 to 5 curving over the striated lamina. We observed electron-dense plaques adjacent to the kinetosomes, electron-dense bodies, numerous free ribosomes, aggregations of glycogen, bowl-shaped membranous structures, rough endoplasmic reticulum, and a novel distinctive pyriform sac of densely packed free ribosomes at the posterior of the cell (8-shaped in transverse section, and delineated by furrowed endoplasmic reticulum). We now propose to rename H. salmonis sensu Ferguson (1979) Spironucleus salmonis. Our review of the morphology of Octomitus salmonis illustrated by Moore (1922a,b) and Davis (1926), has shown that this organism also belongs to the genus Spironucleus. We synonymise H. salmonis sensu Ferguson (1979) with O. salmonis sensu Moore (1922) and Davis (1926), and rename them S. salmonis. An expanded diagnostic guide, including new cytoplasmic elements, is presented for the 4 species of Spironucleus from fishes (S. barkhanus, S. salmonis, S. torosa and S. vortens). We recommend that type descriptions of diplomonads be based on organisms taken directly from the host rather

  20. High-throughput screening of tick-borne pathogens in Europe

    DEFF Research Database (Denmark)

    Michelet, Lorraine; Delannoy, Sabine; Devillers, Elodie

    2014-01-01

    , Bartonella, Candidatus Neoehrlichia, Coxiella, Francisella, Babesia, and Theileria genus) across 94 samples. We successfully determined the prevalence of expected (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Rickettsia helvetica, Candidatus Neoehrlichia mikurensis, Babesia divergens, Babesia...... was conducted on 7050 Ixodes ricinus nymphs collected from France, Denmark, and the Netherlands using a powerful new high-throughput approach. This advanced methodology permitted the simultaneous detection of 25 bacterial, and 12 parasitic species (including; Borrelia, Anaplasma, Ehrlichia, Rickettsia...... venatorum), unexpected (Borrelia miyamotoi) and rare (Bartonella henselae) pathogens in the three European countries. Moreover we detected Borrelia spielmanii, Borrelia miyamotoi, Babesia divergens, and Babesia venatorum for the first time in Danish ticks. This surveillance method represents a major...

  1. Dispersal syndromes related to edge distance in cerrado sensu stricto fragments of central-western Brazil

    Directory of Open Access Journals (Sweden)

    André Vitor Fleuri Jardim

    2009-10-01

    Full Text Available The main selective forces affecting the fruiting strategies are related to the environment in which plants occur. As a savanna, microclimatic conditions should not vary in relation to distance from edge in cerrado sensu stricto fragments. Thus, we postulated that the importance of different dispersal syndromes would not vary towards the fragment core. Our aim was to test in four cerrado sensu stricto fragments in central Brazil whether the absolute density of anemo-, auto-, and zoochorous individuals varied in relation to edge distance. According to results, the absolute density of anemo-and autochorous individuals did not vary, whereas those of zoochorous individuals increased with edge distance, pointing out that there were other factors rather than abiotic conditions shifting zoochorous species to the interior of cerrado sensu stricto fragments.As principais forças seletivas que afetam as estratégias de frutificação estão relacionadas ao ambiente em que as plantas ocorrem. Como fragmentos de cerrado sensu stricto são savânicos, as condições microclimáticas não devem variar em relação à distância da borda. Assim, postulamos que a importância das diferentes síndromes de dispersão não varia da borda ao interior de um fragmento de vegetação. O objetivo deste trabalho foi testar, em quatro fragmentos de cerrado sensu stricto (centro-oeste do Brasil, se a densidade absoluta de indivíduos anemo, auto e zoocóricos variava em função da distância da borda. Segundo nossos resultados, a densidade absoluta de indivíduos anemo e autocóricos não variou significativamente borda em direção ao interior dos fragmentos, enquanto que a dos indivíduos zoocóricos aumentou, indicando que existem outros fatores, que não as condições abióticas, deslocando as espécies zoocóricas para o interior dos fragmentos de cerrado sensu stricto.

  2. Influences of Host Community Characteristics on Borrelia burgdorferi Infection Prevalence in Blacklegged Ticks

    Science.gov (United States)

    Chiu, Grace S.; Smouse, Peter E.; Fonseca, Dina M.; Brisson, Dustin; Morin, Peter J.; Ostfeld, Richard S.

    2017-01-01

    Lyme disease is a major vector-borne bacterial disease in the USA. The disease is caused by Borrelia burgdorferi, and transmitted among hosts and humans, primarily by blacklegged ticks (Ixodes scapularis). The ~25 B. burgdorferi genotypes, based on genotypic variation of their outer surface protein C (ospC), can be phenotypically separated as strains that primarily cause human diseases—human invasive strains (HIS)—or those that rarely do. Additionally, the genotypes are non-randomly associated with host species. The goal of this study was to examine the extent to which phenotypic outcomes of B. burgdorferi could be explained by the host communities fed upon by blacklegged ticks. In 2006 and 2009, we determined the host community composition based on abundance estimates of the vertebrate hosts, and collected host-seeking nymphal ticks in 2007 and 2010 to determine the ospC genotypes within infected ticks. We regressed instances of B. burgdorferi phenotypes on site-specific characteristics of host communities by constructing Bayesian hierarchical models that properly handled missing data. The models provided quantitative support for the relevance of host composition on Lyme disease risk pertaining to B. burgdorferi prevalence (i.e. overall nymphal infection prevalence, or NIPAll) and HIS prevalence among the infected ticks (NIPHIS). In each year, NIPAll and NIPHIS was found to be associated with host relative abundances and diversity. For mice and chipmunks, the association with NIPAll was positive, but tended to be negative with NIPHIS in both years. However, the direction of association between shrew relative abundance with NIPAll or NIPHIS differed across the two years. And, diversity (H') had a negative association with NIPAll, but positive association with NIPHIS in both years. Our analyses highlight that the relationships between the relative abundances of three primary hosts and the community diversity with NIPAll, and NIPHIS, are variable in time and

  3. Bread, beer and wine: yeast domestication in the Saccharomyces sensu stricto complex.

    Science.gov (United States)

    Sicard, Delphine; Legras, Jean-Luc

    2011-03-01

    Yeasts of the Saccharomyces sensu stricto species complex are able to convert sugar into ethanol and CO(2) via fermentation. They have been used for thousands years by mankind for fermenting food and beverages. In the Neolithic times, fermentations were probably initiated by naturally occurring yeasts, and it is unknown when humans started to consciously add selected yeast to make beer, wine or bread. Interestingly, such human activities gave rise to the creation of new species in the Saccharomyces sensu stricto complex by interspecies hybridization or polyploidization. Within the S. cerevisiae species, they have led to the differentiation of genetically distinct groups according to the food process origin. Although the evolutionary history of wine yeast populations has been well described, the histories of other domesticated yeasts need further investigation.

  4. 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.

  5. Identification of new compounds with high activity against stationary phase Borrelia burgdorferi from the NCI compound collection.

    Science.gov (United States)

    Feng, Jie; Shi, Wanliang; Zhang, Shuo; Zhang, Ying

    2015-06-03

    Lyme disease is the leading tick-borne disease in the USA. Whereas the majority of Lyme disease patients with early disease can be cured with standard treatment, some patients suffer from chronic fatigue and joint and muscular pain despite treatment, a syndrome called posttreatment Lyme disease syndrome. Although the cause is unclear, ineffective killing of Borrelia burgdorferi persisters by current Lyme disease antibiotics is one possible explanation. We took advantage of our recently developed high-throughput viability assay and screened the National Cancer Institute compound library collection consisting of 2526 compounds against stationary phase B. burgdorferi. We identified the top 30 new active hits, including the top six anthracycline antibiotics daunomycin 3-oxime, dimethyldaunomycin, daunomycin, NSC299187, NSC363998 and nogalamycin, along with other compounds, including prodigiosin, mitomycin, nanaomycin and dactinomycin, as having excellent activity against B. burgdorferi stationary phase culture. The anthracycline or anthraquinone compounds, which are known to have both anti-cancer and antibacterial activities, also had high activity against growing B. burgdorferi with low minimum inhibitory concentration. Future studies on the structure-activity relationship and mechanisms of action of anthracyclines/anthraquinones are warranted. In addition, drug combination studies with the anthracycline class of compounds and the current Lyme antibiotics to eradicate B. burgdorferi persisters in vitro and in animal models are needed to determine if they improve the treatment of Lyme disease.

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

    Science.gov (United States)

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

    2015-01-01

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

  7. 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

  8. Differentiation of females in Sergentomyia sensu stricto (Diptera: Psychodidae) using scanning electron microscopy of pharyngeal armatures.

    Science.gov (United States)

    Benabdennbi, I; Bombard, S; Braverman, Y; Pesson, B

    1996-03-01

    Scanning electron microscopy of external ornamentation and internal armature of the pharynx was used to identify females of Sergentomyia sensu stricto. Five species from the eastern Mediterranean basin were compared; S. minuta clearly was separated from species of the fallax-group. Within the fallax-group, S. fallax was distinguished readily by its heart-shaped pharynx and the difference in armature between the dorsal and lateral plates.

  9. Effectiveness of disinfectants used in hemodialysis against both Candida orthopsilosis and C. parapsilosis sensu stricto biofilms.

    Science.gov (United States)

    Pires, Regina Helena; da Silva, Julhiany de Fátima; Gomes Martins, Carlos Henrique; Fusco Almeida, Ana Marisa; Pienna Soares, Christiane; Soares Mendes-Giannini, Maria José

    2013-05-01

    Biofilms have been observed in the fluid pathways of hemodialysis machines. The impacts of four biocides used for the disinfection of hemodialysis systems were tested against Candida parapsilosis sensu stricto and Candida orthopsilosis biofilms generated by isolates obtained from a hydraulic circuit that were collected in a hemodialysis unit. Acetic acid was shown to be the most effective agent against Candida biofilms. Strategies for effective disinfection procedures used for hemodialysis systems should also seek to kill and inhibit biofilms.

  10. Discrimination between E. granulosus sensu stricto, E. multilocularis and E. shiquicus Using a Multiplex PCR Assay

    OpenAIRE

    Cong-Nuan Liu; Zhong-Zi Lou; Li Li; Hong-Bin Yan; David Blair; Meng-Tong Lei; Jin-Zhong Cai; Yan-Lei Fan; Jian-Qiu Li; Bao-Quan Fu; Yu-Rong Yang; McManus, Donald P; Wan-Zhong Jia

    2015-01-01

    Background Infections of Echinococcus granulosus sensu stricto (s.s), E. multilocularis and E. shiquicus are commonly found co-endemic on the Qinghai-Tibet plateau, China, and an efficient tool is needed to facilitate the detection of infected hosts and for species identification. Methodology/Principal Findings A single-tube multiplex PCR assay was established to differentiate the Echinococcus species responsible for infections in intermediate and definitive hosts. Primers specific for E. gra...

  11. A Escola Fiocruz de Governo na profissionalização de servidores públicos federais: uma análise dos cursos lato sensu para a Anvisa.

    Directory of Open Access Journals (Sweden)

    Cecília Andrade de Melo e SILVA

    2015-09-01

    Full Text Available O trabalho teve como objetivo a análise quanti-qualitativa dos Trabalhos de Conclusão-TCC do “Curso de Especialização em Vigilância Sanitária dos Servidores da Anvisa” entre 2006 e 2009, cujo principal objeto era formar servidores recém-ingressos da Agencia Nacional de Vigilância Sanitária – Anvisa no campo da saúde coletiva. Foram examinados 366 TCC, a partir de seus resumos. Foram divididos os trabalhos por Macroeixos e categorizados entre artigos, projetos de pesquisa e monografias. Os resultados demonstraram que os temas que mais predominaram como objeto de estudo foram “vigilância” e “medicamentos”.

  12. Isolation and drug susceptibility of Candida parapsilosis sensu lato and other species of C. parapsilosis complex from patients with blood stream infections and proposal of a novel LAMP identification method for the species.

    Science.gov (United States)

    Trabasso, Plinio; Matsuzawa, Tetsuhiro; Fagnani, Renata; Muraosa, Yasunori; Tominaga, Kenichiro; Resende, Mariangela Ribeiro; Kamei, Katsuhiko; Mikami, Yuzuru; Schreiber, Angelica Zaninelli; Moretti, Maria Luiza

    2015-02-01

    Candida parapsilosis complex (CPC) is the third Candida species isolated in blood cultures of patients from our Hospital, following C. albicans and C. tropicalis. From 2006 to 2010, the median annual distribution of CPC was 8 cases/year. Records of 36 patients were reviewed. CPC were 31 (86.1%) C. parapsilosis; 4 (11.1%) C. orthopsilosis; and 1 (2.8%) C. metapsilosis. Clinical characteristics were central venous catheter, 34 (94.4%); parental nutrition, 25 (70%); surgery, 27 (57.9%); prior bacteremia, 20 (51.3%); malignancy, 18 (50%). General mortality was 47.2%. Death was higher in immunosuppressed patients (17 vs. 11; p = 0.003). Three out four (75%) patients with C. orthopsilosis and 14 out 31 (45.2%) with C. parapsilosis died (p = 0.558). Thirty-nine individual isolates were tested for susceptibility to seven antifungal drugs, with MICs values showing susceptibility to all of them. Two isolates, one C. orthopsilosis and one C. parapsilosis, had fluconazole MIC = 4 μg/mL. Differentiation among CPC has implication in caring for patients with invasive candidiasis since there are differences in virulence, pathogenicity and drug susceptibility. A method targeting the topoisomerase II gene based on loop-mediated isothermal amplification (LAMP) was developed. LAMP emerges as a promising tool for the identification of fungal species due to the high sensitivity and specificity. LAMP can be performed at the point-of-care, being no necessary the use of expensive equipment. In our study, the method was successful comparing to the DNA sequencing and proved to be a reliable and fast assay to distinguish the three species of CPC.

  13. Halocoryza Alluaud 1919, sea-side beetles of the Indian, Atlantic (sensu lato, and Pacific Oceans: a generic synopsis and description of a remarkable new species from Baja California Sur, México (Coleoptera, Carabidae, Scaritini, Clivinina

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    Terry Erwin

    2011-09-01

    Full Text Available Information on the three previously described species of Halocoryza Alluaud is updated and a new species for the genus from Isla Carmen, Sea of Cortés, Baja California Sur, México is described. Halocoryza whiteheadiana sp. n. was found at UV light on a beach of that island. This species does not fit the profile of the other three species, i.e., living on coralline beach sands, or in the Mangrove intertidal zone. Two alternative possibilities as to why this is so are suggested and a study plan for testing these possibilities is proposed.

  14. A new genus of athecate interstitial dinoflagellates, Togula gen. nov., previously encompassed within Amphidinium sensu lato: Inferred from light and electron microscopy and phylogenetic analyses of partial large subunit ribosomal DNA sequences

    DEFF Research Database (Denmark)

    Jørgensen, Mårten Flø; Murray, Shauna; Daugbjerg, Niels

    2004-01-01

    and scanning electron microscopy did not reveal significant differences between the vegetative motile cells; however, cells about to undergo mitosis developed longitudinal grooves on the hypocone in one of the clades but not in the other. Both clades differed substantially from A. britannicum in partial large...

  15. A novel dodine-free selective medium based on the use of cetyl trimethyl ammonium bromide (CTAB) to isolate Beauveria bassiana, Metarhizium anisopliae sensu lato and Paecilomyces lilacinus from soil.

    Science.gov (United States)

    Posadas, Julieta B; Comerio, Ricardo M; Mini, Jorge I; Nussenbaum, Ana L; Lecuona, Roberto E

    2012-01-01

    This study evaluated the quaternary ammonium compound cetyl trimethyl ammonium bromide (CTAB) as an alternative to the chemically related dodecylguanidine (dodine) for the selective isolation of entomopathogenic fungi. Oatmeal agar (OA) with chloramphenicol was used as basal medium, and three concentrations of CTAB (0.5, 0.6, 0.7 g/L) were evaluated and compared against OA + 0.46 g/L dodine. Selective isolation and growth studies were performed with the entomopathogens Beauveria bassiana, Metarhizium anisopliae s.l. and Paecilomyces lilacinus and five common non-entomopathogenic non-target species. The three entomopathogenic fungi sporulated earlier on OA + 0.6 g/L CTAB than on OA + 0.46 g/L dodine, while none of the non-target fungi sporulated on OA + 0.6 g/L CTAB. All entomopathogenic fungal isolates grew on OA + 0.6 g/L CTAB, despite some intra-species variation, whereas non-target fungi showed no growth or sporulation. OA + 0.6 g/L CTAB resulted in an efficient medium to isolate B. bassiana, M. anisopliae s. l. and P. lilacinus from soil samples. Results of our study suggest that OA + 0.6 g/L CTAB is a suitable, simple and inexpensive to prepare medium to replace OA + 0.46 g/L dodine for the selective isolation of these fungi.

  16. [Determination of antibodies against Borrelia burgdorferi in patients with morphea, lichen sclerosus et atrophicus and erythema chronicum migrans].

    Science.gov (United States)

    Pinazo Canales, I; Betlloch Mas, I; Mestre Bauza, F; Salva Armengod, F; Parras Vázquez, F; Alomar Cardell, J

    1990-04-01

    Several cutaneous entities described in Europe as Chronic Migrans Erythema (CME), Mild Cutis Lymphadenosis (MCL) and Chronic Atrophyc Acrodermatitis (CAA) constitute clinical manifestations of a Borrellia Burgdorferi. The presence of clinical and hystologic lesions similar to those of liquen esclerosus and atrophyc (LEA) and localized esclerodermia (morphea) in patients with CAA has driven to several authors to demonstrate the aethiologic participation of B. Burgdorferi in patients carrying those cutaneous lesions with contradictory results. A serologic study with indirect immunofluorescence (IIF) and FIAX test was performed in 16 patients (9 with morphea, 6 with LEA and 1 with CME) in order to evaluate the role of this microorganism in our environment. Five reactive sera were obtained of which only one presented IgG antibodies titrated at 1/256. The IgM antibodies by IIF and IgG by FIAX test turned out to be negative. A specific relationship between B. Burgdorferi and the studied entities could not be established.

  17. The incidence of Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti coinfections among foresters and farmers in eastern Poland

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    Anna Pañczuk

    2016-01-01

    Full Text Available Background & objectives: Lyme borreliosis is the most common tick-borne disease in Europe and the USA. However, a great variety of pathogens are transmitted by ticks, which results in mixed infections, with Lyme borreliosis. The aim of the present study was to show the incidence of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti coinfections among the population of foresters and farmers, as these people, due to their profession, are particularly exposed to tick contact. Methods: The study was carried out in eastern Poland (the northern part of the Lublin Province in 2013. The study was performed in a group of 93 individuals occupationally exposed to tick bites (foresters and farmers, whose blood serum showed the presence of IgG anti-B. burgdorferi antibodies. Blood serum in this group were evaluated for IgG anti-A. phagocytophilum and IgG anti-B. microti antibodies by means of IFA IgG indirect immunofluorescence tests. Information related to age, sex, number of tick bite episodes, presence of various symptoms related to the tick bites, and antibiotic therapy applied as treatment for diagnosed Lyme borreliosis were obtained from the subjects through a structured questionnaire. The results were analyzed in Statistica v. 7.1 statistical analysis software. Results: The presence of IgG antibodies against the analyzed pathogens revealed B. burgdorferi and A. phagocytophilum coinfection in 26 (28% of the examined subjects and B. burgdorferi and B. microti coinfection in one person (1.1%. No coinfection with all the three pathogens was observed in any individual. The co-occurrence of headache plus bone, joint and muscle pain was noted significantly more often among individuals diagnosed with B. burgdorferi and A. phagocytophilum coinfection. Interpretation & conclusion: Foresters and farmers are exposed to B. burgdorferi and A. phagocytophilum coinfection in the study area. Therefore, it is probable that these pathogens may severely

  18. Evaluation of the Importance of VlsE Antigenic Variation for the Enzootic Cycle of Borrelia burgdorferi.

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    Artem S Rogovskyy

    Full Text Available 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 locus for B. burgdorferi persistence. However, studies involving vls mutant clones have thus far only utilized in vitro-grown or host-adapted spirochetes and laboratory strains of mice. Additionally, the effects of vls mutation on tick acquisition and transmission has not yet been tested. Thus, the importance of VlsE antigenic variation for persistent infection of the natural reservoir host, and for the B. burgdorferi enzootic life cycle in general, has not been examined to date. In the current work, Ixodes scapularis and Peromyscus maniculatus were infected with different vls mutant clones to study the importance of the vls locus for the enzootic cycle of the Lyme disease pathogen. The findings highlight the significance of the vls system for long-term infection of the natural reservoir host, and show that VlsE antigenic variability is advantageous for efficient tick acquisition of B. burgdorferi from the mammalian reservoir. The data also indicate that the adaptation state of infecting spirochetes influences B. burgdorferi avoidance from host antibodies, which may be in part due to its respective VlsE expression levels. Overall, the current findings provide the most direct evidence on the importance of VlsE for the enzootic cycle of Lyme disease spirochetes, and underscore the significance of VlsE antigenic variation for maintaining B. burgdorferi in nature.

  19. Evaluation of the Importance of VlsE Antigenic Variation for the Enzootic Cycle of Borrelia burgdorferi.

    Science.gov (United States)

    Rogovskyy, Artem S; Casselli, Timothy; Tourand, Yvonne; Jones, Cami R; Owen, Jeb P; Mason, Kathleen L; Scoles, Glen A; Bankhead, Troy

    2015-01-01

    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 locus for B. burgdorferi persistence. However, studies involving vls mutant clones have thus far only utilized in vitro-grown or host-adapted spirochetes and laboratory strains of mice. Additionally, the effects of vls mutation on tick acquisition and transmission has not yet been tested. Thus, the importance of VlsE antigenic variation for persistent infection of the natural reservoir host, and for the B. burgdorferi enzootic life cycle in general, has not been examined to date. In the current work, Ixodes scapularis and Peromyscus maniculatus were infected with different vls mutant clones to study the importance of the vls locus for the enzootic cycle of the Lyme disease pathogen. The findings highlight the significance of the vls system for long-term infection of the natural reservoir host, and show that VlsE antigenic variability is advantageous for efficient tick acquisition of B. burgdorferi from the mammalian reservoir. The data also indicate that the adaptation state of infecting spirochetes influences B. burgdorferi avoidance from host antibodies, which may be in part due to its respective VlsE expression levels. Overall, the current findings provide the most direct evidence on the importance of VlsE for the enzootic cycle of Lyme disease spirochetes, and underscore the significance of VlsE antigenic variation for maintaining B. burgdorferi in nature.

  20. Identification of tick-borne pathogens in ticks feeding on humans in Turkey.

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    Ömer Orkun

    2014-08-01

    Full Text Available The importance of tick-borne diseases is increasing all over the world, including Turkey. The tick-borne disease outbreaks reported in recent years and the abundance of tick species and the existence of suitable habitats increase the importance of studies related to the epidemiology of ticks and tick-borne pathogens in Turkey. The aim of this study was to investigate the presence of and to determine the infection rates of some tick-borne pathogens, including Babesia spp., Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in the ticks removed from humans in different parts of Ankara.A total of 169 ticks belonging to the genus Haemaphysalis, Hyalomma, Ixodes and Rhipicephalus were collected by removing from humans in different parts of Ankara. Ticks were molecularly screened for Babesia spp., Borrelia burgdorferi sensu lato and spotted fever group rickettsiae by PCR and sequencing analysis. We detected 4 Babesia spp.; B. crassa, B. major, B. occultans and B. rossi, one Borrelia spp.; B. burgdorferi sensu stricto and 3 spotted fever group rickettsiae; R. aeschlimannii, R. slovaca and R. hoogstraalii in the tick specimens analyzed. This is the report showing the presence of B. rossi in a region that is out of Africa and in the host species Ha. parva. In addition, B. crassa, for which limited information is available on its distribution and vector species, and B. occultans, for which no conclusive information is available on its presence in Turkey, were identified in Ha. parva and H. marginatum, respectively. Two human pathogenic rickettsia species (R. aeschlimannii and R. slovaca were detected with a high prevalence in ticks. Additionally, B. burgdorferi sensu stricto was detected in unusual tick species (H. marginatum, H. excavatum, Hyalomma spp. (nymph and Ha. parva.This study investigates both the distribution of several tick-borne pathogens affecting humans and animals, and the presence of new tick-borne pathogens in Turkey

  1. 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

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

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    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

  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. Elastase is the only human neutrophil granule protein that alone is responsible for in vitro killing of Borrelia burgdorferi.

    Science.gov (United States)

    Garcia, R; Gusmani, L; Murgia, R; Guarnaccia, C; Cinco, M; Rottini, G

    1998-04-01

    Phagocytosis of Borrelia burgdorferi by human polymorphonuclear leukocytes triggers oxygen-dependent and -independent mechanisms of potentially cidal outcome. Nevertheless, no factor or process has yet been singled out as being borreliacidal. We have studied the B. burgdorferi-killing ability of the myeloperoxidase-H2O2-chloride system and that of primary and secondary granule components in an in vitro assay. We found that neither secondary granule acid extracts nor the chlorinating system could kill these microorganisms, while primary granule extracts were effective. The Borrelia-killing factor was purified to homogeneity and demonstrated to be elastase. Its cidal activity was found to be independent of its proteolytic activity.

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

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    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

  6. [Detection of antibodies of Borrelia burgdorferi among inhabitants of north-eastern Poland].

    Science.gov (United States)

    Pancewicz, S A; Januszkiewicz, A; Hermanowska-Szpakowicz, T

    1996-01-01

    The aim of this work was to estimate the detection of antibodies against Borrelia burgdorferi among inhabitants in North Eastern Poland 1765 persons from Białystok, Lomza and Suwałki voivodships were examined. Among them 1101 persons were from high-risk of exposition to ticks group (forest workers, people living close to forests). 418 (23.68%) persons from group of 1765 had antibodies against B.b. There was no difference of incidence of antibodies against B.b. in high-risk group and the others. The results show that North Eastern Poland is the endemic region of occurrence of B.b.

  7. Prevalence of Borrelia burgdorferi-infected ticks from wildlife hosts, a response to Norris et al.

    OpenAIRE

    Esteve-Gassent, Maria D.; Grover, Abha; Teresa P. Feria-Arroyo; Castro-Arellano, Ivan; Medina, Raul F; Gordillo-Pérez, Guadalupe; de León, Adalberto A Pérez

    2015-01-01

    In a recent Letter to the Editor, Norris et al. questioned the validity of some of our data reported by Feria-Arroyo et al. The main issue investigated by us was the potential impact of climate change on the probable distribution of the tick vector Ixodes scapularis in the Texas-Mexico transboundary region. As an ancillary issue, an analysis of sequence data for the intergenic spacer of Borrelia burgdorferi was conducted. In the present letter, we provide further evidence supporting our origi...

  8. Diversidade alfa e beta no cerrado sensu strictu da Chapada Pratinha, Brasil Alfa and Beta diversity in the cerrado sensu stricto

    Directory of Open Access Journals (Sweden)

    Maria Cristina Felfili

    2001-08-01

    Full Text Available O bioma cerrado possui uma das mais ricas floras dentre as savanas mundiais com mais de 6000 espécies, abrange uma vasta extensão territorial, contém as três maiores bacias hidrográficas sul americanas, e se destaca pela elevada biodiversidade. O objetivo deste trabalho, que faz parte do Projeto Biogeografia do Bioma Cerrado, foi analisar as diversidades alfa e beta em comunidades de cerrado sensu stricto. Foram amostrados o Parque Nacional de Brasília, a Estação Ecológica de Águas Emendadas, a Area de Proteção Ambiental (APA Gama-Cabeça de Veado, Silvânia-GO, Paracatu-MG e Patrocínio-MG. A amostragem foi aleatória com 10 parcelas de 20x50m em cada área de estudo. Foram incluídas plantas lenhosas, exceto lianas, que tivessem no mínimo 5cm de diâmetro. Foi calculada a diversidade alfa pelo teste de Shannon & Wienner e o de Simpson. A diversidade beta foi calculada pelo índice de Whittaker que mede a mudança ou taxa de substituição na composição de espécies de um local para outro. Foi efetuada a curva espécie-área para as 60 parcelas amostradas e efetuada a classificação por TWINSPAN. O cerrado sensu stricto da Chapada Pratinha pode ser diferenciado em duas zonas fitogeográficas: Distrito Federal-Silvânia e Paracatu-Patrocínio. Estas coincidem com um zoneamento por sistemas de terra que classifica a primeira zona como terras altas em contraposição à segunda.The cerrado flora is one of the richest among the world's savannas with more than 6000 species. The cerrado covers a large territory, contains the three most important hydrographycal basins in South America and a high diversity. The objective of this work, that is part of the project Biogeography of the Cerrado Biome, was to analyse alfa and beta diversity in the cerrado sensu stricto. The sampled places were: Brasília National Park -- DF, Águas Emendadas Ecological Station -- DF, Environmentally protected Area of Gama-Cabeça de Veado -- DF, Silv

  9. 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.

  10. 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

  11. Seasonal distribution of Borreliae in Ixodes ricinus ticks in the Belgrade region

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    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.

  12. [Lyme disease].

    Science.gov (United States)

    Portillo, Aránzazu; Santibáñez, Sonia; Oteo, José A

    2014-02-01

    Lyme disease (LD) is a worldwide-distributed multisystemic process caused by Borrelia burgdorferi sensu lato (s.l.) and transmitted by hard ticks. In fact, it is the most common tick-borne infectious disease in the northern hemisphere. In Spain it is transmitted by Ixodes ricinus ticks and Borrelia garinii is the genoespecies of B. burgdorferi s.l. mostly involved in our area. LD is known as "the last great imitator" due to the broad clinical spectrum that may cause. Except in the case of erythema migrans (pathognomonic feature of the disease), the remaining clinical manifestations should be confirmed using microbiological tests. This review is intended to provide readers a current vision of the etiology, epidemiology, clinical manifestations, laboratory diagnosis and treatment of Lyme disease in our environment. Controversial aspects arising from the use of non-validated microbiological tests that are being used without scientific rigor are highlighted.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. Altered murine tissue colonization by Borrelia burgdorferi following targeted deletion of linear plasmid 17-carried genes.

    Science.gov (United States)

    Casselli, Timothy; Tourand, Yvonne; Bankhead, Troy

    2012-05-01

    The causative agent of Lyme disease, Borrelia burgdorferi, possesses a segmented genome comprised of a single linear chromosome and upwards of 23 linear and circular plasmids. Much of what is known about plasmid-borne genes comes from studying laboratory clones that have spontaneously lost one or more plasmids during in vitro passage. Some plasmids, including the linear plasmid lp17, are never or rarely reported to be lost during routine culture; therefore, little is known about the requirement of these conserved plasmids for infectivity. In this study, the effects of deleting regions of lp17 were examined both in vitro and in vivo. A mutant strain lacking the genes bbd16 to bbd25 showed no deficiency in the ability to establish infection or disseminate to the bloodstream of mice; however, colonization of peripheral tissues was delayed. Despite the ability to colonize ear, heart, and joint tissues, this mutant exhibited a defect in bladder tissue colonization for up to 56 days postinfection. This phenotype was not observed in immunodeficient mice, suggesting that bladder colonization by the mutant strain was inhibited by an adaptive immune-based mechanism. Moreover, the mutant displayed increased expression of outer surface protein C in vitro, which was correlated with the absence of the gene bbd18. To our knowledge, this is the first report involving genetic manipulation of lp17 in an infectious clone of B. burgdorferi and reveals for the first time the effects of lp17 gene deletion during murine infection by the Lyme disease spirochete.

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

    Science.gov (United States)

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

    2016-05-01

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

  19. 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...

  20. 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.

  1. Interleukin-10 (IL-10) inhibits Borrelia burgdorferi-induced IL-17 production and attenuates IL-17-mediated Lyme arthritis.

    Science.gov (United States)

    Hansen, Emily S; Medić, Velinka; Kuo, Joseph; Warner, Thomas F; Schell, Ronald F; Nardelli, Dean T

    2013-12-01

    Previous studies have shown that cells and cytokines associated with interleukin-17 (IL-17)-driven inflammation are involved in the arthritic response to Borrelia burgdorferi infection. Here, we report that IL-17 is a contributing factor in the development of Lyme arthritis and show that its production and histopathological effects are regulated by interleukin-10 (IL-10). Spleen cells obtained from B. burgdorferi-infected, "arthritis-resistant" wild-type C57BL/6 mice produced low levels of IL-17 following stimulation with the spirochete. In contrast, spleen cells obtained from infected, IL-10-deficient C57BL/6 mice produced a significant amount of IL-17 following stimulation with B. burgdorferi. These mice developed significant arthritis, including erosion of the bones in the ankle joints. We further show that treatment with antibody to IL-17 partially inhibited the significant hind paw swelling and histopathological changes observed in B. burgdorferi-infected, IL-10-deficient mice. Taken together, these findings provide additional evidence of a role for IL-17 in Lyme arthritis and reveal an additional regulatory target of IL-10 following borrelial infection.

  2. 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.

  3. Ability of an oral formulation of afoxolaner to protect dogs from Borrelia burgdorferi infection transmitted by wild Ixodes scapularis ticks.

    Science.gov (United States)

    Baker, C F; McCall, J W; McCall, S D; Drag, M D; Mitchell, E B; Chester, S T; Larsen, D

    2016-12-01

    A randomized, blinded, negative controlled study was conducted to determine whether treatment with afoxolaner (NexGard(®), Merial, Inc.) would prevent the transmission of Borrelia burgdorferi to dogs by wild caught Ixodes scapularis ticks. Twenty healthy dogs were randomly assigned to two groups of ten dogs each. Ten dogs were treated orally on Day 0 at a dose near the minimum recommended dose of afoxolaner of 2.5mg/kg (actual doses 2.5-3.1mg/kg) and ten control dogs were not treated. On Day 28, each dog was infested with approximately 50 adult unfed wild caught I. scapularis that had a 67% B. burgdorferi infection rate (determined by polymerase chain reaction). On Day 33, live ticks were counted and removed. No ticks were found on treated dogs while control dogs had an average of 21.4 ticks. To detect infection, the B. burgdorferi-specific C6 antibody SNAP(®) 4Dx(®) test (IDEXX) was performed on serum collected before infestation (all dogs seronegative on Days -6 and 27) and on Days 48, 63, 77 and 92. The ten treated dogs remained seronegative through the end of the study (Day 92), while nine out of the ten control dogs were infected, as demonstrated by their seroconversion to being positive for the presence of the B. burgdorferi-specific C6 antibody starting on Day 48. In this study, all dogs treated with NexGard(®) 28days prior to challenge with wild caught I. scapularis ticks were protected from B. burgdorferi infection, while nine out of the ten untreated control dogs were infected.

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

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    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.

  5. 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.

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

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    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.

  7. Borreliose de Lyme Lyme borreliosis

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    Mônica Santos

    2010-12-01

    Full Text Available As borrelioses constituem um grupo de doenças infecciosas causadas por espiroquetas do gênero Borrelia. A borreliose de Lyme, também denominada doença de Lyme, é uma doença infecciosa, não contagiosa, causada por espiroquetas pertencentes ao complexo Borrelia burgdorferi Sensu Lato e transmitida, mais frequentemente, por picada de carrapatos do gênero Ixodes. A doença apresenta quadro clínico variado, podendo desencadear manifestações cutâneas, articulares, neurológicas e cardíacas.Borreliosis is an infectious disease caused by spirochetes of the genus Borrelia. Lyme borreliosis, also known as Lyme disease, is a non-contagious infectious disease caused by spirochetes belonging to the complex Borrelia burgdorferi sensu lato and more often transmitted by the bite of infected ticks of the genus Ixodes.The disease is characterized by a varied clinical profile, which can trigger cutaneous, articular, neurological and cardiac manifestations.

  8. Analysis of Sporothrix schenckii sensu stricto and Sporothrix brasiliensis virulence in Galleria mellonella.

    Science.gov (United States)

    Clavijo-Giraldo, Diana M; Matínez-Alvarez, José A; Lopes-Bezerra, Leila M; Ponce-Noyola, Patricia; Franco, Bernardo; Almeida, Ricardo S; Mora-Montes, Héctor M

    2016-03-01

    The study of the host-pathogen interaction is essential to understand the mechanisms underlying adhesion, colonization and tissue damage by pathogens. This is usually achieved by performing in vivo studies using small mammals, such as rats, mice and guinea pigs. Nowadays, the mouse models of systemic or subcutaneous infection are the gold standard assays to analyze the virulence of members of the Sporothrix schenckii complex. There are, however, invertebrates that have been recently used as alternative hosts to assess the virulence of both bacteria and fungi, and among them, larvae of Galleria mellonella are popular because they are easy to breed, and require non-specialized facilities to maintain the colony. Here, we assessed the use of G. mellonella larvae to test the virulence of S. schenckii sensu stricto and Sporothrix brasiliensis strains, and found that infection with yeast-like cells, but not with conidia or germlings, reproduces the virulence data generated in the mouse model of infection. Furthermore, with this insect model we could classify the virulence of some strains as low, intermediate or high, in line with the observations in the mammalian model. Therefore, G. mellonella is suitable, and a new alternative, to test virulence of both S. schenckii sensu stricto and S. brasiliensis.

  9. STRICTO SENSU POST-GRADUATION PROGRAMME NURSING: THE OPINION OF UNDERGRADUATES

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    Estela Maria Leite Meirelles Monteiro

    2012-02-01

    Full Text Available Normal 0 21 false false false PT-BR X-NONE X-NONE Objetivo: Conhecer o entendimento dos graduandos de Enfermagem de uma Universidade Pública de Pernambuco sobre a contribuição de um Programa Associado de Pós-Graduação Stricto Sensu em Enfermagem. Métodos: Estudo descritivo-exploratório qualitativo. Foram entrevistados trinta estudantes de enfermagem nos meses de agosto a novembro de 2009. Os dados foram apresentados utilizando-se a técnica do Discurso do Sujeito Coletivo. Resultado: Foram evidenciadas três Idéias Centrais: Conhecimento sobre o Programa Associado de Pós-Graduação Stricto Sensu em Enfermagem da UPE; contribuição para a prática profissional e o interesse em ingressar. Conclusão: O entendimento da proposta de fortalecimento das pesquisas e de uma articulação destas com as tecnologias e práticas profissionais vem requerer um redirecionamento na filosofia das IES, tendo a incumbência de despertar o mais precocemente, entre seus graduandos o interesse e a habilidade por pesquisar, sedimentando sua formação na busca contínua de conhecimentos, na renovação dos saberes, na inquietação por investigar objetos de estudo instrumentalizadores do agir profissional em saúde.

  10. 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.

  11. Fitossociologia de cerrado sensu stricto no município de Abaeté-MG Phytosociology of the cerrado sensu stricto in Abaeté, MG, Brazil

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    Amilcar Walter Saporetti Jr

    2003-06-01

    Full Text Available O cerrado tem sido objeto de discussão de grupos temáticos que estudam a conservação de biodiversidade no Estado de Minas Gerais. São inúmeras as áreas de conservação com vegetação de cerrado das quais não se têm informações a respeito de sua composição florística e estrutura. O objetivo deste trabalho foi avaliar florística e fitossociologicamente uma área de cerrado sensu stricto, no município de Abaeté-MG. A área de estudo é um fragmento com 2 ha de cerrado sensu stricto, preservado como área de reserva da CAF-Santa Bárbara, situada nas coordenadas 19º05'S e 44º58'W, a uma altitude de 480 m, em leve depressão próxima de uma vereda. O clima pertence ao tipo Cwa pelo sistema de Köppen, com precipitação média anual de 1.400 mm. O solo é do tipo Latossolo Vermelho distrófico. Foram instaladas 15 parcelas de 200 m² (10 x 20 m, distribuídas sistematicamente ao longo de trilhas, distanciadas 10 m entre si. Foram amostrados todos os indivíduos lenhosos vivos com circunferência do caule ao nível do solo (CAS igual ou maior que 10 cm. O índice de Shannon foi de 3,590 e a equabilidade foi de 0,804, considerados comuns para cerrados bem conservados. Foram amostrados 1.339 indivíduos, sendo a composição florística constituída por 85 espécies, distribuídas em 44 famílias. As famílias com maior número de espécies foram Leguminosae Caesalpinioideae com sete espécies, Annonaceae com cinco, Myrtaceae, Malpighiaceae, Erythroxylaceae, Anacardiaceae, Rubiaceae e Bignoniaceae com quatro, seguidas de Vochysiaceae e Leguminosae Papilionoideae com três. As espécies que apresentaram o maior valor de importância (VI foram Xylopia aromatica (Lam. Mart. (22,21, Myrcia lingua Berg (18,18 Caryocar brasiliense Cambess. (17,91, Eugenia dysenterica DC. (17,58, Byrsonima intermedia A. Juss. (13,69 e Brosimum gaudichaudii Trécul (11,86.The cerrado has been a topic of discussion of thematic groups studying biodiversity

  12. Prevalence Rates of Borrelia burgdorferi (Spirochaetales: Spirochaetaceae), Anaplasma phagocytophilum (Rickettsiales: Anaplasmataceae), and Babesia microti (Piroplasmida: Babesiidae) in Host-Seeking Ixodes scapularis (Acari: Ixodidae) from Pennsylvania.

    Science.gov (United States)

    Hutchinson, M L; Strohecker, M D; Simmons, T W; Kyle, A D; Helwig, M W

    2015-07-01

    The etiological agents responsible for Lyme disease (Borrelia burgdorferi), human granulocytic anaplasmosis (Anaplasma phagocytophilum), and babesiosis (Babesia microti) are primarily transmitted by the blacklegged tick, Ixodes scapularis Say. Despite Pennsylvania having in recent years reported the highest number of Lyme disease cases in the United States, relatively little is known regarding the geographic distribution of the vector and its pathogens in the state. Previous attempts at climate-based predictive modeling of I. scapularis occurrence have not coincided with the high human incidence rates in parts of the state. To elucidate the distribution and pathogen infection rates of I. scapularis, we collected and tested 1,855 adult ticks statewide from 2012 to 2014. The presence of I. scapularis and B. burgdorferi was confirmed from all 67 Pennsylvania counties. Analyses were performed on 1,363 ticks collected in the fall of 2013 to avoid temporal bias across years. Infection rates were highest for B. burgdorferi (47.4%), followed by Ba. microti (3.5%) and A. phagocytophilum (3.3%). Coinfections included B. burgdorferi+Ba. microti (2.0%), B. burgdorferi+A. phagocytophilum (1.5%) and one tick positive for A. phagocytophilum+Ba. microti. Infection rates for B. burgdorferi were lower in the western region of the state. Our findings substantiate that Lyme disease risk is high throughout Pennsylvania.

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

    Directory of Open Access Journals (Sweden)

    JOPPERT Adriana Marques

    2001-01-01

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

  14. 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

  15. [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

  16. Co-feeding transmission facilitates strain coexistence in Borrelia burgdorferi, the Lyme disease agent.

    Science.gov (United States)

    States, S L; Huang, C I; Davis, S; Tufts, D M; Diuk-Wasser, M A

    2016-12-26

    Coexistence of multiple tick-borne pathogens or strains is common in natural hosts and can be facilitated by resource partitioning of the host species, within-host localization, or by different transmission pathways. Most vector-borne pathogens are transmitted horizontally via systemic host infection, but transmission may occur in the absence of systemic infection between two vectors feeding in close proximity, enabling pathogens to minimize competition and escape the host immune response. In a laboratory study, we demonstrated that co-feeding transmission can occur for a rapidly-cleared strain of Borrelia burgdorferi, the Lyme disease agent, between two stages of the tick vector Ixodes scapularis while feeding on their dominant host, Peromyscus leucopus. In contrast, infections rapidly became systemic for the persistently infecting strain. In a field study, we assessed opportunities for co-feeding transmission by measuring co-occurrence of two tick stages on ears of small mammals over two years at multiple sites. Finally, in a modeling study, we assessed the importance of co-feeding on R0, the basic reproductive number. The model indicated that co-feeding increases the fitness of rapidly-cleared strains in regions with synchronous immature tick feeding. Our results are consistent with increased diversity of B. burgdorferi in areas of higher synchrony in immature feeding - such as the midwestern United States. A higher relative proportion of rapidly-cleared strains, which are less human pathogenic, would also explain lower Lyme disease incidence in this region. Finally, if co-feeding transmission also occurs on refractory hosts, it may facilitate the emergence and persistence of new pathogens with a more limited host range.

  17. Seroprevalence of Borrelia burgdorferi in occupationally exposed persons in the Belgrade area, Serbia

    Directory of Open Access Journals (Sweden)

    Dragutin Jovanovic

    2015-09-01

    Full Text Available Lyme disease (LD is a natural focal zoonotic disease caused by Borrelia burgdorferi, which is mainly transmitted through infected Ixodes ricinus tick bites. The presence and abundance of ticks in various habitats, the infectivity rate, as well as prolonged human exposure to ticks are factors that may affect the infection risk as well as the incidence of LD. In recent years, 20% to 25% of ticks infected with different borrelial species, as well as about 5,300 citizens with LD, have been registered in the Belgrade area. Many of the patients reported tick bites in city’s grassy areas. The aim of this study was to assess the seroprevalence of B. burgdorferi in high-risk groups (forestry workers and soldiers in the Belgrade area, and to compare the results with healthy blood donors. A two-step algorithm consisting of ELISA and Western blot tests was used in the study. Immunoreactivity profiles were also compared between the groups. The results obtained showed the seroprevalence to be 11.76% in the group of forestry workers, 17.14% in the group of soldiers infected by tick bites and 8.57% in the population of healthy blood donors. The highest IgM reactivity was detected against the OspC protein, while IgG antibodies showed high reactivity against VlsE, p19, p41, OspC, OspA and p17. Further investigations in this field are necessary in humans and animals in order to improve protective and preventive measures against LD.

  18. Seroprevalence of Borrelia burgdorferi in occupationally exposed persons in the Belgrade area, Serbia

    Science.gov (United States)

    Jovanovic, Dragutin; Atanasievska, Sonja; Protic-Djokic, Vesna; Rakic, Uros; Lukac-Radoncic, Elvira; Ristanovic, Elizabeta

    2015-01-01

    Lyme disease (LD) is a natural focal zoonotic disease caused by Borrelia burgdorferi, which is mainly transmitted through infected Ixodes ricinus tick bites. The presence and abundance of ticks in various habitats, the infectivity rate, as well as prolonged human exposure to ticks are factors that may affect the infection risk as well as the incidence of LD. In recent years, 20% to 25% of ticks infected with different borrelial species, as well as about 5,300 citizens with LD, have been registered in the Belgrade area. Many of the patients reported tick bites in city’s grassy areas. The aim of this study was to assess the seroprevalence of B. burgdorferi in high-risk groups (forestry workers and soldiers) in the Belgrade area, and to compare the results with healthy blood donors. A two-step algorithm consisting of ELISA and Western blot tests was used in the study. Immunoreactivity profiles were also compared between the groups. The results obtained showed the seroprevalence to be 11.76% in the group of forestry workers, 17.14% in the group of soldiers infected by tick bites and 8.57% in the population of healthy blood donors. The highest IgM reactivity was detected against the OspC protein, while IgG antibodies showed high reactivity against VlsE, p19, p41, OspC, OspA and p17. Further investigations in this field are necessary in humans and animals in order to improve protective and preventive measures against LD. PMID:26413064

  19. 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

  20. Individual and environmental factors associated with the seroprevalence of Borrelia burgdorferi in Belgian farmers and veterinarians

    Science.gov (United States)

    De Keukeleire, Mathilde; Robert, Annie; Kabamba, Benoît; Dion, Elise; Luyasu, Victor; Vanwambeke, Sophie O.

    2016-01-01

    Background Lyme disease (LD) is a common tick-borne disease in Europe. Diverse factors at various scales determine the spatial distribution of Borrelia burgdorferi infection risk and a better understanding of those factors in a spatially explicit framework is needed for disease management and prevention. While the ecology of ticks and the landscape favoring their abundance have been extensively studied, the environmental conditions favoring an intense contact with susceptible humans, including groups at risk, are sparse. The aim of this study is to assess which individual and environmental factors can favor B. burgdorferi infection in a Belgian group professionally at risk. Methods Serological results of 127 veterinarians and farmers enrolled in this study were analyzed, taking into account their municipality of residence. Using binary logistic regression and considering interaction terms, the joint effects of landscape composition and configuration, and forest and wildlife management were examined. Results Seven of the 127 workers were seropositive for LD, leading to a seroprevalence of 5.51%. Seropositivity was higher in older persons. The proportion of forest and semi-natural habitats and wetland had a positive impact on LD seroprevalence while arable land–grassland ecotones had a negative one. Our results confirmed the need to consider complex interactions between landscape variables in order to model risk. Conclusions Our data show that LD has to be considered as a risk for farmers and veterinarians. Rather than focusing either on ecological aspects of tick and pathogen distribution or on purely epidemiological aspects such as individual risk factors, our model highlights the role of human–environment interactions in LD risk assessment. PMID:27852421

  1. 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

  2. [The production of knowledge regarding gestational hypertension in the stricto sensu graduate nursing studies in Brazil].

    Science.gov (United States)

    Martins, Marialda; Monticelli, Marisa; Brüggemann, Odaléa Maria; Costa, Roberta

    2012-08-01

    This documental study analyzed the Brazilian production of stricto sensu nursing graduate programs related to gestational hypertension. The data source used was the Brazilian Nursing Association Theses and Dissertations Database. The survey located 14 studies produced between 1979-2008, produced mainly in the Southeast region between 1996 and2008. The analytical process revealed a concern with the subjectivity of pregnant women and with aspects regarding nursing care. Furthermore, it showed that most studies used a qualitative methodology supported by nursing theories. The experience lived by pregnant women with hypertension is marked by negative feelings and socioeconomic problems, and is also affected by how the family is organized. The culture of pregnant women with hypertension is disregarded and they receive care in a context in which the disease is the priority. In conclusion, despite some scientific advancements, this topic has not raised the interest it deserves among nurses attending graduate study programs.

  3. Sporothrix schenckii sensu stricto isolated from soil in an armadillo's burrow.

    Science.gov (United States)

    Rodrigues, Anderson Messias; Bagagli, Eduardo; de Camargo, Zoilo Pires; Bosco, Sandra de Moraes Gimenes

    2014-04-01

    Sporotrichosis is a polymorphic disease of man and animals caused by traumatic implantation of propagules into the skin and subcutaneous tissue. Pathogenic species includes S. brasiliensis, S. schenckii, S. globosa and S. luriei. The disease is remarkable for its occurrence as sapronoses and/or zoonosis outbreaks in tropical and subtropical areas; although, the ecology of the clinical clade is still puzzling. Here, we describe an anamorphic Sporothrix strain isolated from soil in an armadillo's burrow, which was located in a hyper endemic area of Paracoccidioidomycosis in Brazil. This isolate was identified as S. schenckii sensu stricto (Clade IIa) based on morphological and physiological characteristics and phylogenetic analyses of calmodulin sequences. We then discuss the role of the nine-banded armadillo Dasypus novemcinctus as a natural carrier of Sporothrix propagules to better understand Sporothrix sources in nature and reveal essential aspects about the pathogen's eco-epidemiology.

  4. Pollen Morphology of Eight Genera of the Subtribe Mutisiinae Less. sensu Bremer (Compositae) from Asia

    Institute of Scientific and Technical Information of China (English)

    Na-Na LIN; Hong WANG; De-Zhu LI; Stephen BLACKMORE

    2005-01-01

    The pollen morphology of 28 species and one variety representing eight genera of the subtribe Mutisiinae Less. sensu Bremer (Compositae) was investigated by scanning electron microscopy (SEM) and light microscopy. The pollen grains are usually radically symmetrical, isopolar, and tricolporate. The shape varies from spheroidal to subprolate in equatorial view and is three-lobed circular in polar view. Based on exine sculpturing observed under SEM, the pollen grains can be divided into three types: (i) type Ⅰ, with macrogranulate sculpture; (ii) type Ⅱ, with a finely granulate sculpture; and (iii) type Ⅲ, with spiny sculpture.The palynological data showed that the Macroclinidium Maxim. was remarkably distinguished from those of the other seven genera. The variation of pollen characteristics within the Mutisiinae in Asia is little at species level, but it proves some useful information for studying the relationships among genera and it has an important significance in further understanding the evolutionary history of Mutisiinae.

  5. A resiliência humana no ambiente acadêmico de cursos stricto sensu

    OpenAIRE

    2015-01-01

    Este estudo surgiu do interesse de compreender de que maneira se dá a questão da resiliência no ambiente acadêmico de pós-graduação stricto sensu, no qual os indivíduos são confrontados com demandas de alta dedicação e esforço que exigem a conformação a um novo modelo de realização de atividades. Como estratégia de pesquisa utilizou-se o estudo comparativo de casos, mediante método de coleta qualitativa empírica, com corte temporal transversal. Selecionaram-se duas antigas e conceituadas inst...

  6. Echinococcus canadensis (G7) and Echinococcus granulosus sensu stricto (G1) in swine of southern Brazil.

    Science.gov (United States)

    Monteiro, D U; Botton, S A; Tonin, A A; Azevedo, M I; Graichen, D A S; Noal, C B; de la Rue, M L

    2014-05-28

    The cystic echinococcosis (CE) is an important zoonotic disease caused by the parasite Echinococcus spp. In Brazil, this parasite is present in Rio Grande do Sul (RS) state, border with Argentina and Uruguay, causing several damages to human and animal health. This study aimed to identify Echinococcus spp. in hydatid cysts of swine and evaluate the similarity of the genotypes through the phylogenetic analysis. A total of 3,101,992 swine were slaughtered in the central/northern region of RS/Brazil, during 2008-2012. Five isolates were characterized as hydatid cyst by molecular analysis, based on the mitochondrial gene cytochrome c oxidase subunit I (cox-I). The genotypes E. granulosus sensu stricto (G1) (n=2) and E. canadensis (G7) (n=3) were identified in the hydatid cysts. The swine represents a potential intermediate host for different genotypes of Echinococcus spp., besides it can contribute to the perpetuation of the parasite's life cycle in rural areas.

  7. ULTRASTRUCTURE OF GYMNODINIUM AUREOLUM (DINOPHYCEAE): TOWARD A FURTHER REDEFINITION OF GYMNODINIUM SENSU STRICTO

    DEFF Research Database (Denmark)

    Hansen, Gert

    2001-01-01

    of Gymnodinium, G. fuscum (Ehrenberg) Stein and certain other Gymnodinium species. The nuclear pores were restricted to these chambers. In the flagellar apparatus a nuclear fibrous connective linked the longitudinal microtubular root and the nucleus. This structure had so far been observed only in Gymnodinium...... spp. and in the heterotrophic species Actiniscus pentasterias (Ehrenberg) Ehrenberg, Nematodinium armatum (Dogiel) Kofoid et Swezy and Polykrikos kofoidii Chatton. Another unusual feature of G. aureolum was the presence of a striated fiber in the longitudinal flagellum, a feature previously only found...... in Ceratium furca (Ehrenberg) Claparède et Lachmann and C. tripos (O.F. Müller) Nitzsch. Gymnodinium aureolum also possessed a prominent ventral protrusion associated with the peduncle and containing electron opaque material. It is concluded that G. aureolum belongs to the Gymnodinium sensu stricto group...

  8. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  10. 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

  11. 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.

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

    OpenAIRE

    2016-01-01

    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...

  13. Borrelia burgdorferi EbfC defines a newly-identified, widespread family of bacterial DNA-binding proteins.

    Science.gov (United States)

    Riley, Sean P; Bykowski, Tomasz; Cooley, Anne E; Burns, Logan H; Babb, Kelly; Brissette, Catherine A; Bowman, Amy; Rotondi, Matthew; Miller, M Clarke; DeMoll, Edward; Lim, Kap; Fried, Michael G; Stevenson, Brian

    2009-04-01

    The Lyme disease spirochete, Borrelia burgdorferi, encodes a novel type of DNA-binding protein named EbfC. Orthologs of EbfC are encoded by a wide range of bacterial species, so characterization of the borrelial protein has implications that span the eubacterial kingdom. The present work defines the DNA sequence required for high-affinity binding by EbfC to be the 4 bp broken palindrome GTnAC, where 'n' can be any nucleotide. Two high-affinity EbfC-binding sites are located immediately 5' of B. burgdorferi erp transcriptional promoters, and binding of EbfC was found to alter the conformation of erp promoter DNA. Consensus EbfC-binding sites are abundantly distributed throughout the B. burgdorferi genome, occurring approximately once every 1 kb. These and other features of EbfC suggest that this small protein and its orthologs may represent a distinctive type of bacterial nucleoid-associated protein. EbfC was shown to bind DNA as a homodimer, and site-directed mutagenesis studies indicated that EbfC and its orthologs appear to bind DNA via a novel alpha-helical 'tweezer'-like structure.

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

    Directory of Open Access Journals (Sweden)

    Amy J Ullmann

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

  15. Detection of Babesia microti and Borrelia burgdorferi in host-seeking Ixodes scapularis (Acari: Ixodidae) in Monmouth County, New Jersey.

    Science.gov (United States)

    Schulze, Terry L; Jordan, Robert A; Healy, Sean P; Roegner, Vivien E

    2013-03-01

    The etiological agents that cause human babesiosis (Babesia microti) and Lyme disease (Borrelia burgdorferi) share a common tick vector (Ixodes scapularis Say) and rodent reservoir (Peromyscus leucopus), but because the geographical distribution of babesiosis is more restricted than Lyme disease, it was not considered a nationally notifiable disease until 2011. Although recent studies have shown dramatic increases in the number of cases of babesiosis and expansion of its range, little is known about infection and coinfection prevalence of these pathogens in the primary tick vector. Of the 478 I. scapularis nymphs collected within six Monmouth County, NJ, municipalities between 2004 and 2006, 4.0 and 10.0% were infected with B. microti and B. burgdorferi, respectively, while 2.9% were coinfected. Analysis of the 610 I. scapularis adults collected during the same period yielded an infection prevalence of 8.2% for B. microti and 45.2% for B. burgdorferi, while 6.2% were coinfected. The potential public health importance of these findings is discussed.

  16. Function of the Borrelia burgdorferi FtsH Homolog Is Essential for Viability both In Vitro and In Vivo and Independent of HflK/C

    Directory of Open Access Journals (Sweden)

    Chen-Yi Chu

    2016-04-01

    Full Text Available In many bacteria, the FtsH protease and its modulators, HflK and HflC, form a large protein complex that contributes to both membrane protein quality control and regulation of the cellular response to environmental stress. Both activities are crucial to the Lyme disease pathogen Borrelia burgdorferi, which depends on membrane functions, such as motility, protein transport, and cell signaling, to respond to rapid changes in its environment. Using an inducible system, we demonstrate that FtsH production is essential for both mouse and tick infectivity and for in vitro growth of B. burgdorferi. FtsH depletion in B. burgdorferi cells resulted in membrane deformation and cell death. Overproduction of the protease did not have any detectable adverse effects on B. burgdorferi growth in vitro, suggesting that excess FtsH does not proteolytically overwhelm its substrates. In contrast, we did not observe any phenotype for cells lacking the protease modulators HflK and HflC (ΔHflK/C, although we examined morphology, growth rate, growth under stress conditions, and the complete mouse-tick infectious cycle. Our results demonstrate that FtsH provides an essential function in the life cycle of the obligate pathogen B. burgdorferi but that HflK and HflC do not detectably affect FtsH function.

  17. In Vitro Activities of Amphotericin B, Terbinafine, and Azole Drugs against Clinical and Environmental Isolates of Aspergillus terreus Sensu Stricto

    Science.gov (United States)

    Fernández, Mariana S.; Rojas, Florencia D.; Cattana, María E.; Sosa, María de los Ángeles; Iovannitti, Cristina A.; Giusiano, Gustavo E.

    2015-01-01

    The antifungal susceptibilities of 40 clinical and environmental isolates of A. terreus sensu stricto to amphotericin B, terbinafine, itraconazole, and voriconazole were determined in accordance with CLSI document M38-A2. All isolates had itraconazole and voriconazole MICs lower than epidemiologic cutoff values, and 5% of the isolates had amphotericin B MICs higher than epidemiologic cutoff values. Terbinafine showed the lowest MICs. No significant differences were found when MICs of clinical and environmental isolates were compared. PMID:25824228

  18. A review of ceratocystis sensu stricto with special reference to the species complexes c. coerulescens and c. fimbriata.

    OpenAIRE

    Marín Montoya, Mauricio; Wingfield, Michael J.

    2011-01-01

    El género Ceratocystis sensu stricto incluye un alto número de hongos fitopatógenos de plantas angiospermas y gimnospermas en diversas regiones del mundo. Entre las principales enfermedades causadas por estos hongos se destacan los marchitamientos vasculares, manchado de maderas, chancros y pudriciones radiculares, de tallos y frutos. Muchas especies producen metabolitos volátiles con olores que atraen una amplia variedad de insectos a los tejidos vegetales infectados, sin embargo, el grado d...

  19. Genetic diversity and population genetic structure analysis of Echinococcus granulosus sensu stricto complex based on mitochondrial DNA signature.

    Directory of Open Access Journals (Sweden)

    Monika Sharma

    Full Text Available The genetic diversity and population genetics of the Echinococcus granulosus sensu stricto complex were investigated based on sequencing of mitochondrial DNA (mtDNA. Total 81 isolates of hydatid cyst collected from ungulate animals from different geographical areas of North India were identified by sequencing of cytochrome c oxidase subunit1 (coxi gene. Three genotypes belonging to E. granulosus sensu stricto complex were identified (G1, G2 and G3 genotypes. Further the nucleotide sequences (retrieved from GenBank for the coxi gene from seven populations of E. granulosus sensu stricto complex covering 6 continents, were compared with sequences of isolates analysed in this study. Molecular diversity indices represent overall high mitochondrial DNA diversity for these populations, but low nucleotide diversity between haplotypes. The neutrality tests were used to analyze signatures of historical demographic events. The Tajima's D test and Fu's FS test showed negative value, indicating deviations from neutrality and both suggested recent population expansion for the populations. Pairwise fixation index was significant for pairwise comparison of different populations (except between South America and East Asia, Middle East and Europe, South America and Europe, Africa and Australia, indicating genetic differentiation among populations. Based on the findings of the present study and those from earlier studies, we hypothesize that demographic expansion occurred in E. granulosus after the introduction of founder haplotype particular by anthropogenic movements.

  20. Erythema chronicum migrans and salmon fishing in Alaska: an enigma resolved by microbiology laboratory

    Directory of Open Access Journals (Sweden)

    Silvio Daffara

    2010-03-01

    Full Text Available A 50-year-old man, pentathlete and fond of salmon fishing, after returning from Alaska was referred to our Division for an erythematous and indolent lesion on the right thigh gradually enlarging from the right knee to the right rear and the buttock.The patient was clinically diagnosed with figurate erythema. Laboratory analysis demonstrated a moderate leukocytosis and hypergammaglobulinemia, accompanied by serological markers of past contact with noted EBV and CMV.A “punch” biopsy of the edge of the lesion showed a superficial and deep perivascular and interstitial infiltrate of lymphocytes, eosinophiles and a few plasma cells, consistent with a borrelial infection. Lyme serology (IFI suggested a questionable borderline positivity; subsequent investigations by ELISA and Western Blot were both positive, leading to the diagnosis of erythema chronicum migrans in course of borreliosis.The diagnosis was further confirmed by positive PCR for Borrelia burgdorferi sensu lato. The erythema resolved after a 2-week doxycycline treatment (2x100 mg die. The amplification product (16S rDNA gene obtained from skin biopsy was sequenced using standard ABI technology, and confirmed the identification of a member of B. burgdorferi sensu lato (sl complex. Since this sequence was not useful to identify the genospecies, further studies were conducted employing a nested PCR targeted on the internal transcribed spacer (ITS of B. burgdorferi, following a protocol previously described (1. The ITS sequence showed 100% identity with B. afzelii, a species not reported from North America, thus prompting us to conclude that the patient was not infected by B. afzelii during the fishing trip in Alaska.

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

    Science.gov (United States)

    Ginsberg, Howard S.; Buckley, P.A.; Balmforth, Maxon G.; Zhioua, Elyes; Mitra, Shaibal; Buckley, Francine G.

    2005-01-01

    Reservoir competence for 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 mist netting on Fire Island, NY, were held in a field laboratory 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 direct fluorescent-antibody staining after molting to the nymphal stage. American robins were competent reservoirs, infecting 16.1% of larvae applied to wild-caught birds, compared with 0% of control ticks placed on uninfected laboratory mice. Robins that were previously infected in the laboratory 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 with birds infected in the laboratory suggest that infected birds display temporal variability in infectiousness to larval ticks. Engorged larvae drop from birds abundantly during daylight, so the abundance of these bird species in the peridomestic environment suggests that they might contribute infected ticks to lawns and gardens.

  2. Changes in Borrelia burgdorferi ELISA antibody over time in both antibiotic treated and untreated horses.

    Science.gov (United States)

    Divers, Thomas J; Grice, Amy L; Mohammed, Hussni O; Glaser, Amy L; Wagner, Bettina

    2012-12-01

    Changes in ELISA serology are frequently used to determine antibiotic treatment success for Lyme disease in horses. This concept was based upon a previous report showing a marked decline in ELISA values in experimentally infected and antibiotic-treated ponies. Changes in Lyme serology following antibiotic treatment in naturally infected horses have not been reported. The objective of this study was to compare Borrelia ELISA antibody concentrations in naturally exposed horses both before and following antibiotic treatment for Lyme disease. A retrospective study was performed comparing oxytetracycline- or doxycyclinetreated (n = 68) and untreated (n = 183) horses from a single equine practice and their change in Borrelia ELISA values over a similar time period. Antibiotictreated horses had a decline in ELISA values in comparison to control horses (P ≤ 0.05) and untreated horses were twice as likely to have their ELISA values increase (OR = 0.5; 95% C.I. = 0.3-0.9) compared to treated horses. The magnitude of the decline in ELISA units following treatments was small compared to that previously reported in experimentally infected and treated ponies. Field-exposed horses with high Borrelia burgdorferi ELISA values who are treated with either oxytetracycline or doxycycline can be expected to have only a small decline in ELISA values following treatment. Persistently high ELISA titres following appropriate treatments for Lyme disease may not, without appropriate clinical signs, be a reason for more prolonged treatment.

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

    Directory of Open Access Journals (Sweden)

    Dan Drecktrah

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

  4. Structure and immunomodulatory property relationship in NapA of Borrelia burgdorferi.

    Science.gov (United States)

    Codolo, Gaia; Papinutto, Elena; Polenghi, Alessandra; D'Elios, Mario Milco; Zanotti, Giuseppe; de Bernard, Marina

    2010-12-01

    NapA from Borrelia burgdorferi is a member of the Dps-like protein family with specific immunomodulatory properties; in particular, NapA is able to induce the expression of IL-23 in neutrophils and monocytes, as well as the expression of IL-6, IL-1β, and transforming growth factor beta (TGF-β) in monocytes, via Toll-like receptor (TLR) 2. Such an activity on innate immune cells triggers a synovial fluid Th17 response. Here we report the crystal structure of NapA, determined at 2.6Å resolution, which shows that the quaternary structure of the protein is that of a dodecamer with 23 symmetry, typical of the proteins of the family. We also demonstrate that the N- and C-terminal tails, which are flexible and not visible in the crystal, are not relevant for its pro-Th17 activity. Based on the crystal structure and on the comparison with the structure of the orthologous protein from Helicobacter pylori, HP-NAP, we hypothesize that the charge distributions on the two proteins' surfaces are responsible for the interaction with TLR2 and for the different behaviors in modulating the immune response.

  5. Cerebrospinal fluid T-regulatory cells recognize Borrelia burgdorferi NAPA in chronic Lyme borreliosis.

    Science.gov (United States)

    Amedei, A; Codolo, G; Ozolins, D; Ballerini, C; Biagioli, T; Jaunalksne, I; Zilevica, A; D Elios, S; De Bernard, M; D' Elios, M M

    2013-01-01

    The NapA protein of B. burgdorferi is essential for the persistence of spirochetes in ticks. One of the most intriguing aspects of NapA is its potential to interfere with the host immune system. Here, we investigated the role of the acquired immune responses induced by NapA in the cerebrospinal fluids (CSF) of patients with chronic Lyme borreliosis. We evaluated the cytokine profile induced in microglia cells and CSF T cells following NapA stimulation. We report here that NapA induced a regulatory T (Treg) response in the CSF of patients with chronic Lyme borreliosis and it is able to expand this suppressive response by promoting the production of TGF-beta and IL-10 by microglia cells. Collectively, these data strongly support a central role of NapA in promoting both Treg response and immune suppression in the CSF of patients with chronic Lyme borreliosis and suggest that NapA and the Treg pathway may represent novel therapeutic targets for the prevention and treatment of the disease.

  6. Helicobacter heilmannii sensu stricto-related gastric ulcers: a case report.

    Science.gov (United States)

    Matsumoto, Takehisa; Kawakubo, Masatomo; Akamatsu, Taiji; Koide, Naohiko; Ogiwara, Naoko; Kubota, Seiko; Sugano, Mitsutoshi; Kawakami, Yoshiyuki; Katsuyama, Tsutomu; Ota, Hiroyoshi

    2014-03-28

    A spiral bacterium (SH9), morphologically different from Helicobacter pylori (H. pylori), was found in a 62-year-old woman's gastric mucosa. Gastroscopic examination revealed multiple gastric ulcers near the pyloric ring; mapping gastric biopsy showed mild mononuclear infiltration with large lymphoid follicles in the antrum, without corpus atrophy. Urea breath test and H. pylori culture were negative, but Giemsa staining of biopsies revealed tightly coiled bacteria that immunostained with anti-H. pylori antibody. Sequencing of SH9 16S rRNA and the partial urease A and B subunit genes showed that the former sequence had highest similarity (99%; 1302/1315 bp) to Helicobacter heilmannii (H. heilmannii) sensu stricto (H. heilmannii s.s.) BC1 obtained from a bobcat, while the latter sequence confirmed highest similarity (98.3%; 1467/1493 bp) to H. heilmannii s.s. HU2 obtained from a human. The patient was diagnosed with multiple gastric ulcers associated with H. heilmannii s.s. infection. After triple therapy (amoxicillin, clarithromycin, and lansoprazole) with regimen for eradicating H. pylori, gastroscopy showed ulcer improvement and no H. heilmannii s.s. upon biopsy.

  7. Echinococcus granulosus Sensu Stricto in Dogs and Jackals from Caspian Sea Region, Northern Iran

    Directory of Open Access Journals (Sweden)

    Shirzad GHOLAMI

    2016-10-01

    Full Text Available Background: The aim of the present study was genotyping of Echinococcus granulosus isolates from dogs and jackals in Mazandaran Province, northern Iran, and using partial sequence of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1.Methods: E. granulosus isolates (n = 15 were collected from 42 stray dogs and 16 jackals found in south of the Caspian Sea in northern Iran. After morphological study, the isolates were genetically characterized using consensus sequences (366bp of the cox1 gene. Phylogenetic analysis of cox1 nucleotide sequence data was performed using a Bayesian Inference approach.Results: Four different sequences were observed among the isolates. Two genotypes [G1 (66.7% and G3 (33.3%] were identified among the isolates. The G1 sequences indicated three sequence profiles. One profile (Maz1 had 100% homology with reference sequence (AN: KP339045. Two other profiles, designated Maz2 and Maz3, had 99% homology with the G1 genotype (ANs: KP339046 and KP339047. A G3 sequence designated Maz4 showed 100% homology with a G3 reference sequence (AN: KP339048.Conclusion: The occurrence of the G1 genotype of E. granulosus sensu stricto as a frequent genotype in dogs is emphasized. This study established the first molecular characterization of E. granulosus in the province.

  8. Echinococcus equinus and Echinococcus granulosus sensu stricto from the United Kingdom: genetic diversity and haplotypic variation.

    Science.gov (United States)

    Boufana, Belgees; Lett, Wai San; Lahmar, Samia; Buishi, Imad; Bodell, Anthony J; Varcasia, Antonio; Casulli, Adriano; Beeching, Nicholas J; Campbell, Fiona; Terlizzo, Monica; McManus, Donald P; Craig, Philip S

    2015-02-01

    Cystic echinococcosis is endemic in Europe including the United Kingdom. However, information on the molecular epidemiology of Echinococcus spp. from the United Kingdom is limited. Echinococcus isolates from intermediate and definitive animal hosts as well as from human cystic echinococcosis cases were analysed to determine species and genotypes within these hosts. Echinococcus equinus was identified from horse hydatid isolates, cysts retrieved from captive UK mammals and copro-DNA of foxhounds and farm dogs. Echinococcus granulosus sensu stricto (s.s.) was identified from hydatid cysts of sheep and cattle as well as in DNA extracted from farm dog and foxhound faecal samples, and from four human cystic echinococcosis isolates, including the first known molecular confirmation of E. granulosus s.s. infection in a Welsh sheep farmer. Low genetic variability for E. equinus from various hosts and from different geographical locations was detected using the mitochondrial cytochrome c oxidase subunit 1 gene (cox1), indicating the presence of a dominant haplotype (EQUK01). In contrast, greater haplotypic variation was observed for E. granulosus s.s. cox1 sequences. The haplotype network showed a star-shaped network with a centrally placed main haplotype (EgUK01) that had been reported from other world regions.

  9. 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...... antigen and the C6 antigen, respectively. Graphical methods to display the antibody response and to choose thresholds for a rise in relative antibody reactivity are shown and discussed. In conclusion, 5.4% of people with tick bites showed a rise in Borrelia-specific antibodies above the 2.5% threshold...

  10. 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.

  11. 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.

  12. Absence of sodA Increases the Levels of Oxidation of Key Metabolic Determinants of Borrelia burgdorferi.

    Science.gov (United States)

    Esteve-Gassent, Maria D; Smith, Trever C; Small, Christina M; Thomas, Derek P; Seshu, J

    2015-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease, alters its gene expression in response to environmental signals unique to its tick vector or vertebrate hosts. B. burgdorferi carries one superoxide dismutase gene (sodA) capable of controlling intracellular superoxide levels. Previously, sodA was shown to be essential for infection of B. burgdorferi in the C3H/HeN model of Lyme disease. We employed two-dimensional electrophoresis (2-DE) and immunoblot analysis with antibodies specific to carbonylated proteins to identify targets that were differentially oxidized in the soluble fractions of the sodA mutant compared to its isogenic parental control strain following treatment with an endogenous superoxide generator, methyl viologen (MV, paraquat). HPLC-ESI-MS/MS analysis of oxidized proteins revealed that several proteins of the glycolytic pathway (BB0057, BB0020, BB0348) exhibited increased carbonylation in the sodA mutant treated with MV. Levels of ATP and NAD/NADH were reduced in the sodA mutant compared with the parental strain following treatment with MV and could be attributed to increased levels of oxidation of proteins of the glycolytic pathway. In addition, a chaperone, HtpG (BB0560), and outer surface protein A (OspA, BBA15) were also observed to be oxidized in the sodA mutant. Immunoblot analysis revealed reduced levels of Outer surface protein C (OspC), Decorin binding protein A (DbpA), fibronectin binding protein (BBK32), RpoS and BosR in the sodA mutant compared to the control strains. Viable sodA mutant spirochetes could not be recovered from both gp91/phox-⁄- and iNOS deficient mice while borrelial DNA was detected in multiple tissues samples from infected mice at significantly lower levels compared to the parental strain. Taken together, these observations indicate that the increased oxidation of select borrelial determinants and reduced levels of critical pathogenesis-associated lipoproteins contribute to the in vivo deficit of the sod

  13. Physiological behaviour of gliotoxigenic Aspergillus fumigatus sensu stricto isolated from maize silage under simulated environmental conditions.

    Science.gov (United States)

    Alonso, V; Vergara, L Díaz; Aminahuel, C; Pereyra, C; Pena, G; Torres, A; Dalcero, A; Cavaglieri, L

    2015-01-01

    Environmental conditions play a key role in fungal development. During the silage production process, humidity, oxygen availability and pH vary among lactic-fermentation phases and among different silage sections. The aim of this work was to study the physiological behaviour of gliotoxicogenic Aspergillus fumigatus strains isolated from maize silage under simulated natural physicochemical conditions - different water activities (a(W)), temperatures (Tº), pH and oxygen pressure - on the growth parameters (growth rate and lag phase) and gliotoxin production. The silage was made with the harvested whole maize plant that was chopped and used for trench-type silo fabrication. Water activity and pH of the silage samples were determined. Total fungal counts were performed on Dichloran Rose Bengal Chloramphenicol agar and Dichloran 18% Glycerol agar. The morphological identification of A. fumigatus was performed with different culture media and at different growth temperature to observe microscopic and macroscopic characteristics. Gliotoxin production by A. fumigatus was determined by HPLC. All strains isolated were morphologically identified as A. fumigatus. Two A. fumigatus strains isolated from the silage samples were selected for the ecophysiological study (A. fumigatus sensu stricto RC031 and RC032). The results of this investigation showed that the fungus grows in the simulated natural physicochemical conditions of corn silage and produces gliotoxin. The study of the physiological behaviour of gliotoxigenic A. fumigatus under simulated environmental conditions allowed its behaviour to be predicted in silage and this will in future enable appropriate control strategies to be developed to prevent the spread of this fungus and toxin production that leads to impairment and reduced quality of silage.

  14. First insights into the genetic diversity of Echinococcus granulosus sensu stricto (s.s.) in Serbia.

    Science.gov (United States)

    Debeljak, Zoran; Boufana, Belgees; Interisano, Maria; Vidanovic, Dejan; Kulisic, Zoran; Casulli, Adriano

    2016-06-15

    Cystic echinococcosis (CE) caused by Echinococcus granulosus sensu stricto (s.s.) is a cosmopolitan zoonotic infection which is endemic in Serbia where it is subject to mandatory reporting. However, information on the incidence of the disease in humans and prevalence of hydatid infection in livestock remains limited. We used sequenced data of the cytochrome c oxidase subunit 1 (cox 1) mitochondrial gene to examine the genetic diversity and population structure of E. granulosus (s.s.) from intermediate hosts from Serbia. We also compared our generated nucleotide sequences with those reported for neighbouring European countries. Echinococcus canadensis was molecularly confirmed from pig and human hydatid isolates. E. granulosus (G1) was confirmed from sheep and cattle hydatid isolates as well as the first molecular confirmation in Serbia of E. granulosus G2 in sheep and E. granulosus G3 in sheep and cattle hydatid isolates. The Serbian E. granulosus (s.s.) parsimony network displayed 2 main haplotypes (SB02 and SB05) which together with the neutrality indices were suggestive of bottleneck and/or balancing selection. Haplotype analysis showed the presence of the common E. granulosus haplotype described from other worldwide regions. Investigation of the pairwise fixation (Fst) index suggested that Serbian populations of E. granulosus (s.s.) from sheep and cattle hosts showed moderate genetic differentiation. Six of the Serbian haplotypes (SB02-SB07) were shared with haplotypes from Bulgaria, Hungary and/or Romania. Further studies using a larger number of hydatid isolates from various locations across Serbia will provide more information on the genetic structure of E. granulosus (s.s.) within this region.

  15. Comparative transcriptomics reveal host-specific nucleotide variation in entomophthoralean fungi

    DEFF Research Database (Denmark)

    de Fine Licht, Henrik Hjarvard; Jensen, Annette Bruun; Eilenberg, Jørgen

    2017-01-01

    -off influencing the capacity to infect alternate hosts. Here, we investigate host adaptation in two morphologically indistinguishable and closely related obligate specialist insect-pathogenic fungi from the phylum Entomophthoromycota, Entomophthora muscae sensu stricto and E. muscae sensu lato, pathogens...

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

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    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.

  17. 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.

  18. Comparison of Five Diagnostic Modalities for Direct Detection of Borrelia burgdorferi in Patients with Early Lyme Disease

    Science.gov (United States)

    Liveris, Dionysios; Schwartz, Ira; McKenna, Donna; Nowakowski, John; Nadelman, Robert; DeMarco, Joseph; Iyer, Radha; Bittker, Susan; Cooper, Denise; Holmgren, Diane; Wormser, Gary P.

    2012-01-01

    Lyme disease, the most commonly reported tick-borne infection in North America, is caused by infection with the spirochete Borrelia burgdorferi. Although an accurate clinical diagnosis can often be made based on the presence of erythema migrans, in research studies microbiologic or molecular microbiologic confirmation of the diagnosis may be required. In this study, we evaluated the sensitivity of five direct diagnostic methods (culture and nested PCR of a 2 mm skin biopsy specimen, nested PCR and quantitative PCR (qPCR) performed on the same 1 mL aliquot of plasma and a novel qPCR-blood culture method) in 66 untreated adult patients with erythema migrans. One or more these tests were positive in 93.9% of the patients. Culture was more sensitive than PCR for both skin and blood, but the difference was only statistically significant for blood samples (p< 0.005). Blood culture was significantly more likely to be positive in patients with multiple erythema migrans skin lesions compared to those with a single lesion (p=0.001). Positive test results among the 48 patients for whom all five assays were performed invariably included either a positive blood or skin culture. The results of this study demonstrate that direct detection methods such as PCR and culture are highly sensitive in untreated adult patients with erythema migrans. This enabled microbiologic or molecular microbiologic confirmation of the diagnosis of B. burgdorferi infection in all but four (6.1%) of the 66 patients evaluated. PMID:22571973

  19. 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.

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

    Science.gov (United States)

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

    2014-01-01

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

  1. Pós-graduação Stricto Sensu em Enfermagem no Brasil: avanços e perspectivas

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    Carmen Gracinda Silvan Scochi

    2013-09-01

    Full Text Available A Enfermagem é campo de conhecimento específico e prática social que se consolida e se fortalece como ciência. No Brasil, tem se desenvolvido em função do aumento e qualificação dos programas de pós-graduação stricto sensu. Este artigo tem como objetivo apresentar um resgate histórico da Pós-Graduação Stricto Sensu de Enfermagem no Brasil e refletir sobre sua evolução, avanços, desafios e perspectivas futuras. Explora-se a criação dos cursos de pós-graduação stricto sensu, destacando os movimentos para se construir a cultura da pós-graduação acadêmica e profissional na Enfermagem. Apresenta-se a trajetória histórica da sua consolidação, expansão, conquista da excelência e visibilidade internacional ao longo de quatro décadas e apontam-se os desafios e as perspectivas futuras. Constata-se que a pós-graduação na área vem contribuindo para o avanço e consolidação do conhecimento científico, tecnológico e de inovação em Enfermagem e saúde, tendo como filosofia, o respeito à diversidade e o livre debate de ideias, a elevação da qualidade de vida e saúde e a efetivação do exercício da cidadania.

  2. 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.

  3. 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.

  4. 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.

  5. A high-throughput genetic screen identifies previously uncharacterized Borrelia burgdorferi genes important for resistance against reactive oxygen and nitrogen species

    Science.gov (United States)

    Ramsey, Meghan E.; Hyde, Jenny A.; Medina-Perez, Diana N.; Gao, Lihui; Lundt, Maureen E.; Li, Xin; Norris, Steven J.

    2017-01-01

    Borrelia burgdorferi, the causative agent of Lyme disease in humans, is exposed to reactive oxygen and nitrogen species (ROS and RNS) in both the tick vector and vertebrate reservoir hosts. B. burgdorferi contains a limited repertoire of canonical oxidative stress response genes, suggesting that novel gene functions may be important for protection of B. burgdorferi against ROS or RNS exposure. Here, we use transposon insertion sequencing (Tn-seq) to conduct an unbiased search for genes involved in resistance to nitric oxide, hydrogen peroxide, and tertiary-butyl hydroperoxide in vitro. The screens identified 66 genes whose disruption resulted in increased susceptibility to at least one of the stressors. These genes include previously characterized mediators of ROS and RNS resistance (including components of the nucleotide excision repair pathway and a subunit of a riboflavin transporter), as well as novel putative resistance candidates. DNA repair mutants were among the most sensitive to RNS in the Tn-seq screen, and survival assays with individual Tn mutants confirmed that the putative ribonuclease BB0839 is involved in resistance to nitric oxide. In contrast, mutants lacking predicted inner membrane proteins or transporters were among the most sensitive to ROS, and the contribution of three such membrane proteins (BB0017, BB0164, and BB0202) to ROS sensitivity was confirmed using individual Tn mutants and complemented strains. Further analysis showed that levels of intracellular manganese are significantly reduced in the Tn::bb0164 mutant, identifying a novel role for BB0164 in B. burgdorferi manganese homeostasis. Infection of C57BL/6 and gp91phox-/- mice with a mini-library of 39 Tn mutants showed that many of the genes identified in the in vitro screens are required for infectivity in mice. Collectively, our data provide insight into how B. burgdorferi responds to ROS and RNS and suggests that this response is relevant to the in vivo success of the organism

  6. 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.

  7. Sphaerospora sensu stricto: taxonomy, diversity and evolution of a unique lineage of myxosporeans (Myxozoa).

    Science.gov (United States)

    Bartošová, Pavla; Fiala, Ivan; Jirků, Miloslav; Cinková, Martina; Caffara, Monica; Fioravanti, Maria Letizia; Atkinson, Stephen Douglas; Bartholomew, Jerri Lee; Holzer, Astrid Sibylle

    2013-07-01

    Myxosporeans (Myxozoa) are eukaryotic parasites, primarily of fish, whose classification is in a state of flux as taxonomists attempt to synthesize the traditional morphology-based system with emerging DNA sequence-based phylogenies. The genus Sphaerospora Thélohan, 1892, which includes pathogenic species that cause significant impacts on fisheries and aquaculture, is one of the most polyphyletic taxa and exemplifies the current challenges facing myxozoan taxonomists. The type species, S. elegans, clusters within the Sphaerospora sensu stricto clade, members of which share similar tissue tropism and long insertions in their variable rRNA gene regions. However, other morphologically similar sphaerosporids lie in different branches of myxozoan phylogenetic trees. Herein, we significantly extend taxonomic sampling of sphaerosporids with SSU+LSU rDNA and EF-2 sequence data for 12 taxa including three representatives of the morphologically similar genus Polysporoplasma Sitjà-Bobadilla et Álvarez-Pellitero, 1995. These taxa were sampled from different vertebrate host groups, biogeographic realms and environments. Our phylogenetic analyses and statistical tests of single and concatenated datasets revealed Sphaerospora s. s. as a strongly supported monophyletic lineage, that clustered sister to the whole myxosporean clade (freshwater+marine lineages). Generally, Sphaerospora s. s. rDNA sequences (up to 3.7 kb) are the longest of all myxozoans and indeed metazoans. The sphaerosporid clade has two lineages, which have specific morphological, biological and sequence traits. Lineage A taxa (marine Sphaerospora spp.) have a single binucleate sporoplasm and shorter AT-rich rDNA inserts. Lineage B taxa (freshwater/brackish Sphaerospora spp.+marine/brackish Polysporoplasma spp.) have 2-12 uninucleate sporoplasms and longer GC-rich rDNA inserts. Lineage B has four subclades that correlate with host group and habitat; all Polysporoplasma species, including the type species

  8. Discrimination between E. granulosus sensu stricto, E. multilocularis and E. shiquicus Using a Multiplex PCR Assay.

    Directory of Open Access Journals (Sweden)

    Cong-Nuan Liu

    2015-09-01

    Full Text Available Infections of Echinococcus granulosus sensu stricto (s.s, E. multilocularis and E. shiquicus are commonly found co-endemic on the Qinghai-Tibet plateau, China, and an efficient tool is needed to facilitate the detection of infected hosts and for species identification.A single-tube multiplex PCR assay was established to differentiate the Echinococcus species responsible for infections in intermediate and definitive hosts. Primers specific for E. granulosus, E. multilocularis and E. shiquicus were designed based on sequences of the mitochondrial NADH dehydrogenase subunit 1 (nad1, NADH dehydrogenase subunit 5 (nad5 and cytochrome c oxidase subunit 1 (cox1 genes, respectively. This multiplex PCR accurately detected Echinococcus DNA without generating nonspecific reaction products. PCR products were of the expected sizes of 219 (nad1, 584 (nad5 and 471 (cox1 bp. Furthermore, the multiplex PCR enabled diagnosis of multiple infections using DNA of protoscoleces and copro-DNA extracted from fecal samples of canine hosts. Specificity of the multiplex PCR was 100% when evaluated using DNA isolated from other cestodes. Sensitivity thresholds were determined for DNA from protoscoleces and from worm eggs, and were calculated as 20 pg of DNA for E. granulosus and E. shiquicus, 10 pg of DNA for E. multilocularis, 2 eggs for E. granulosus, and 1 egg for E. multilocularis. Positive results with copro-DNA could be obtained at day 17 and day 26 after experimental infection of dogs with larval E. multilocularis and E. granulosus, respectively.The multiplex PCR developed in this study is an efficient tool for discriminating E. granulosus, E. multilocularis and E. shiquicus from each other and from other taeniid cestodes. It can be used for the detection of canids infected with E. granulosus s.s. and E. multilocularis using feces collected from these definitive hosts. It can also be used for the identification of the Echinococcus metacestode larva in intermediate

  9. CRITERIA OF POSITIVITY FOR Ig ANTIBODIES IN THE METHOD OF IMMUNE BLOTTING OF LYME DISEASE

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    V G Barskova

    2001-01-01

    Full Text Available There are currently no accepted criteria for positive Western blots in Russian patients with Lyme borreliosis. The purpose of the current study was to develop criteria for a positive IgG westem-blot to aid particularly in the diagnosis of patients with joint manifestation of the disorder. Patients: 97 with Lyme disease, 145 - control subjects. IgG antibody responses were determined to 3 species ofB.burgdorferi sensu lato by Western blotting, using blots prepared by manufacturer. The best discriminatory ability of test criteria was chained by requiring any 3 of 11 IgG bands, a definition that could be used with B. burgdorferi sensu stricto, B.garinii and B.afzelii strains. With these 3 antigen preparation, positive IgG blots were found in 0 to 18% of patients with localized erythema migrans of < 4 weeks duration, 23 to 39% of those with disseminated infection < 20 weeks duration, and in 39 to 46% of those with late arthritis/arthralgia of >6 months duration the specificity was 93 to 99%. Thus, IgG Western blotting may bring greater specificity to serologic testing in Lyme borreliosis, but the sensitivity is limited.

  10. Sensitivity of Lyme Borreliosis Spirochetes to Serum Complement of Regular Zoo Animals: Potential Reservoir Competence of Some Exotic Vertebrates.

    Science.gov (United States)

    Ticha, Lucie; Golovchenko, Maryna; Oliver, James H; Grubhoffer, Libor; Rudenko, Nataliia

    2016-01-01

    Reaction of vertebrate serum complement with different Borrelia burgdorferi sensu lato species is used as a basis in determining reservoir hosts among domesticated and wild animals. Borrelia burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii were tested for their sensitivity to sera of exotic vertebrate species housed in five zoos located in the Czech Republic. We confirmed that different Borrelia species have different sensitivity to host serum. We found that tolerance to Borrelia infection possessed by hosts might differ among individuals of the same genera or species and is not affected by host age or sex. Of all zoo animals included in our study, carnivores demonstrated the highest apparent reservoir competency for Lyme borreliosis spirochetes. We showed that selected exotic ungulate species are tolerant to Borrelia infection. For the first time we showed the high tolerance of Siamese crocodile to Borrelia as compared to the other studied reptile species. While exotic vertebrates present a limited risk to the European human population as reservoirs for the causative agents of Lyme borreliosis, cases of incidental spillover infection could lead to successful replication of the pathogens in a new host, changing the status of selected exotic species and their role in pathogen emergence or maintenance. The question if being tolerant to pathogen means to be a competent reservoir host still needs an answer, simply because the majority of exotic animals might never be exposed to spirochetes in their natural environment.

  11. 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

  12. A survey for infection with Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi, and Babesia canis in feral and client-owned dogs in the Turks and Caicos Islands, British West Indies

    OpenAIRE

    Hoff, Brent; McEwen, Beverly; Peregrine, Andrew S.

    2008-01-01

    The frequency of infection with Dirofilaria immitis and Babesia canis and seropositivity to Ehrlichia canis and Borrelia burgdorferi in feral and client-owned dogs was determined. Feral dogs were 14.8 and 11.2 times more likely to be seropositive to D. immitis and E. canis, respectively, than were client-owned dogs. None of the dogs tested positive for B. burgdorferi or B. canis.

  13. Recent study on canine vector-borne zoonoses in southern Slovakia - serologic survey.

    Science.gov (United States)

    Čabanová, Viktória; Pantchev, Nikola; Hurníková, Zuzana; Miterpáková, Martina

    2015-12-01

    Over the last decade a significant spread of Canine Vector Borne Diseases has been recorded in Central Europe. The aim of the study described here, was to collect current data on the occurrence and distribution of three major canine vector-borne pathogens in the veterinary clinical practice by a newly-developed commercial ELISA test for the detection of Dirofilaria immitis antigen as well as specific circulating antibodies to Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato. Circulating D. immitis antigen was detected in five of 180 investigated sera samples. Two of D. immitis seropositive dogs revealed also microfilariae of D. repens in the blood and three of them were negative for the presence of microfilariae in the Knott's test. From the practical point of view, the finding of D. immitis occult infections might influence existing knowledge about distribution of this species among dogs in Central European countries. In 11.7% of the tested dogs the presence of specific antibodies against A. phagocytophilum was confirmed. Antibodies against B. burgdorferi s.l. were detected in 2.8% of tested sera samples. Coinfection with A. phagocytophilum and B. burgdorferi s.l. was observed in two dogs from Košice district in south-eastern Slovakia. Our data point toward the presence of Canine Vector Borne Diseases in the studied area. Therefore, veterinarians should include these diseases in their differential diagnosis and higher awareness should be focused also on prophylactic measures to prevent the pathogens transmission by arthropod vectors.

  14. 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.

  15. Primary embryonic cells of Rhipicephalus microplus and Amblyomma cajennense ticks as a substrate for the development of Borrelia burgdorferi (strain G39/40

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    J. Rezende

    Full Text Available Borrelia burgdorferi, the agent of Lyme borreliosis, is a spirochetes transmitted by ticks to humans and animals. Its cultivation in vitro in tick cells allows studies of its biology and provides methodology for future research in Brazil, and for the isolation of Borrelia spp. We examined in vitro the characteristics of embryonic cells of Rhipicephalus microplus and Amblyomma cajennense in cell culture and investigated the suitability of embryonic cells as a substrate for cultivation of B. burgdorferi. Subcultures were prepared from primary cultures of embrionary cells of R. microplus and A. cajennense maintained in Leibovitz's (L-15 complete medium at 28 ºC and 31 ºC, respectively. When a monolayer had formed, the L-15 was replaced with Barbour-Stoener-Kelly medium for experiments to infect cell cultures with B. burgdorferi. After 72 hours of cultivation, the spirochetes were counted using an inverted phase contrast microscope and dark-field illumination (400×. Survival, multiplication and the adherence of B. burgdorferi for embryonic cells of R. microplus and A. cajennense were observed. B. burgdorferi cultured with embryonic cells of R. microplus grew on average to a density (final count of 2.4 × 10(7 spirochetes/mL, whereas in cell-free culture, an average of 2.5 × 10(7 spirochetes/mL were counted. When cultivated with A. cajennense cells, the final count of spirochetes was on average 1.7 × 10(7 spirochetes/mL, while spirochetes cultured under cell-free conditions replicated on average of 2.2 × 10(7 spirochetes/mL. Similar results were observed in the final count of Spirochetes cultivated in cells of R. microplus and A. cajennense, when compared with cell-free control. These results demonstrated that cells of R. microplus and A. cajennense have the potential to be used as growth substrate for B. burgdorferi in the study of its interaction with host cells.

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

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    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

  17. Role of sand lizards in the ecology of Lyme and other tick-borne diseases in the Netherlands

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    Spitzen - van der Sluijs Annemarieke

    2010-05-01

    Full Text Available Abstract Background Lizards are considered zooprophylactic for almost all Borrelia burgdorferi species, and act as dilution hosts in parts of North America. Whether European lizards significantly reduce the ability of B. burgdorferi to maintain itself in enzootic cycles, and consequently decrease the infection rate of Ixodes ricinus ticks for B. burgdorferi and other tick-borne pathogens in Western Europe is not clear. Results Ticks were collected from sand lizards, their habitat (heath and from the adjacent forest. DNA of tick-borne pathogens was detected by PCR followed by reverse line blotting. Tick densities were measured at all four locations by blanket dragging. Nymphs and adult ticks collected from lizards had a significantly lower (1.4% prevalence of B. burgdorferi sensu lato, compared to questing ticks in heath (24% or forest (19%. The prevalence of Rickettsia helvetica was significantly higher in ticks from lizards (19% than those from woodland (10% whereas neither was significantly different from the prevalence in ticks from heather (15%. The prevalence of Anaplasma and Ehrlichia spp in heather (12% and forest (14% were comparable, but significantly lower in ticks from sand lizards (5.4%. The prevalence of Babesia spp in ticks varied between 0 and 5.3%. Tick load of lizards ranged from 1 - 16. Tick densities were ~ 5-fold lower in the heather areas than in woodlands at all four sites. Conclusions Despite their apparent low reservoir competence, the presence of sand lizards had insignificant impact on the B. burgdorferi s.l. infection rate of questing ticks. In contrast, sand lizards might act as reservoir hosts for R. helvetica. Remarkably, the public health risk from tick-borne diseases is approximately five times lower in heather than in woodland, due to the low tick densities in heather.

  18. 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.

  19. Molecular Identification of Borrelia miyamotoi in Ixodes ricinus from Portugal.

    Science.gov (United States)

    Nunes, Mónica; Parreira, Ricardo; Lopes, Nádia; Maia, Carla; Carreira, Teresa; Sousa, Carmelita; Faria, Sofia; Campino, Lenea; Vieira, M Luísa

    2015-08-01

    Borrelia miyamotoi, a relapsing fever spirochete, has been found recently in Ixodes ricinus ticks; however, little is known about its spatial distribution and potential local impact on human health. A total of 640 ticks (447 nymphs and 193 adults) collected throughout Portugal were analyzed using two nested PCR protocols, one targeting the flagellin gene and the other the internal transcribed space region between the 5S and the 23S rRNA. As a result, B. miyamotoi was detected, for the first time, in one guesting I. ricinus nymph collected in the Lisboa district. In addition, a prevalence of 11% (71/640) for B. burgdorferi sensu lato was obtained. Even though no human relapsing fever cases due to infection by B. miyamotoi have been reported yet in Portugal, surveillance must be improved to provide better insight into the prevalence and distribution of this spirochete in ticks.

  20. Serological survey of Borrelia infection of dogs in Sapporo, Japan, where Borrelia garinii infection was previously detected.

    Science.gov (United States)

    Uesaka, Karin; Maezawa, Masaki; Inokuma, Hisashi

    2016-03-01

    A serological survey of Borrelia infection of dogs was performed in Sapporo, Japan, where Borrelia garinii infection in dogs was detected in 2011. A total of 314 serum samples were collected from dogs that visited three animal hospitals in Sapporo from 2012 to 2014. The two-step evaluation method, involving screening ELISA followed by Western blot analysis, was used to detect antibodies against Borrelia species. A total of 34 samples were positive by ELISA. Among those 34 samples, 32 were positive for Borrelia spp. by Western blot. These findings suggest that the 32 dogs (10.2%) generated antibodies against Borrelia burgdorferi sensu lato, such as B. garinii or B. afzelii. Antibody positivity was 7.6% and 13.3% for dogs living in urban and rural areas, respectively. Dogs with a history of tick infestation showed a positive rate of 16.7%, which was higher, although not significantly, than the 6.7% among dogs without a history.

  1. REVISIÓN DE Ceratocystis sensu stricto CON ESPECIAL REFERENCIA A LOS COMPLEJOS DE LAS ESPECIES C. coerulescens Y C. fimbriata A REVIEW OF Ceratocystis sensu stricto WITH SPECIAL REFERENCE TO THE SPECIES COMPLEXES C. coerulescens AND C. fimbriata

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    Mauricio Marín Montoya

    2006-06-01

    Full Text Available El género Ceratocystis sensu stricto incluye un alto número de hongos fitopatógenos de plantas angiospermas y gimnospermas en diversas regiones del mundo. Entre las principales enfermedades causadas por estos hongos se destacan los marchitamientos vasculares, manchado de maderas, chancros y pudriciones radiculares, de tallos y frutos. Muchas especies producen metabolitos volátiles con olores que atraen una amplia variedad de insectos a los tejidos vegetales infectados, sin embargo, el grado de asociación entre los insectos y las diferentes especies de Ceratocystis es altamente variable, presentandose asociaciones mutualistas tales como aquellas entre C. polonica, C. laricicola y C. rufipenni e insectos escolítidos, así como también relaciones no específicas como en los casos de C. paradoxa, C. fagacearum y C. fimbriata con nitidúlidos, moscas y diversos coleópteros. El alto grado de similitud morfológica encontrada entre estos hongos hace difícil la definición de los límites entre las especies de Ceratocystis. Durante la pasada década, la utilización de estudios filogenéticos basados en comparaciones de DNA ha contribuido sustancialmente a clarificar la taxonomía de este grupo de hongos, así por ejemplo, dichos estudios han permitido concluir que algunas especies de Ceratocystis realmente representan complejos de especies crípticas. Esta revisión tiene como objetivo principal proveer una introducción a los estudios experimentales desarrollados para las especies de Ceratocystis sensu stricto y brevemente discutir aspectos relacionados con los denominados hongos ophiostomatoides.The genus Ceratocystis sensu stricto Ellis & Halst. includes many economically important plant pathogens of both angiosperms and gymnosperms, worldwide. Diseases caused by these fungi include vascular wilts, sap stains on logs and lumber, stem cankers and rots of roots, stems and fruits. Most Ceratocystis species are well adapted to dispersal by

  2. Behavioural responses of Anopheles gambiae sensu stricto to components of human breath, sweat and urine depend on mixture composition and concentration

    NARCIS (Netherlands)

    Qiu, Y.T.; Smallegange, R.C.; Loon, van J.J.A.; Takken, W.

    2011-01-01

    Host-seeking behaviour of the anthropophilic malaria vector Anopheles gambiae sensu stricto (Diptera: Culicidae) is mediated predominantly by olfactory cues. Several hundreds of odour components have been identified from human emanations, but only a few have been proven to act as attractants or syne

  3. The effect of aliphatic carboxylic acids on olfaction-based host-seeking of the malaria mosquito Anopheles gambiae sensu stricto

    NARCIS (Netherlands)

    Smallegange, R.C.; Qiu, Y.T.; Bukovinszkine-Kiss, G.; Loon, van J.J.A.; Takken, W.

    2009-01-01

    The role of aliphatic carboxylic acids in host-seeking response of the malaria mosquito Anopheles gambiae sensu stricto was examined both in a dual-choice olfactometer and with indoor traps. A basic attractive blend of ammonia + lactic acid served as internal standard odor. Single carboxylic acids w

  4. Identification of candidate volatiles that affect the behavioural response of the malaria mosquito Anopheles gambiae sensu stricto to an active kairomone blend: laboratory and semi-field assays

    NARCIS (Netherlands)

    Smallegange, R.C.; Bukovinszkine Kiss, G.; Otieno, B.; Mbadi, P.A.; Takken, W.; Mukabana, W.R.; Loon, van J.J.A.

    2012-01-01

    Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is the most important vector of human malaria in sub-Saharan Africa, affecting the lives of millions of people. Existing tools such as insecticide-treated nets and indoor-residual sprays are not only effective, but also have limitations as a

  5. Synergism between ammonia, lactic acid and carboxylic acids as kairomones in the host-seeking behaviour of the malaria mosquito Anopheles gambiae sensu stricto (Diptera: Culicidae)

    NARCIS (Netherlands)

    Smallegange, R.C.; Qiu, Y.T.; Loon, van J.J.A.; Takken, W.

    2005-01-01

    Host odours play a major role in the orientation and host location of blood-feeding mosquitoes. Anopheles gambiae Giles sensu stricto, which is the most important malaria vector in Africa, is a highly anthropophilic mosquito species, and the host-seeking behaviour of the females of this mosquito is

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

    Directory of Open Access Journals (Sweden)

    Renata S.A. Carlos

    2007-09-01

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

  7. [New records of ectoparasites of the eastern water bat Myotis petax Hollister, 1912 (Vespertilionidae, Chiroptera) and the revision of the previous material collected from Myotis daubentonii s. lato in Eastern Palaearctic].

    Science.gov (United States)

    Orlova, M V; Orlov, O L; Zhigalin, A V

    2014-01-01

    New findings of ectoparasites from the eastern water bat Myotis petax Hollister, 1912, belonging to the Siberian-Russian Far Eastern complex and earlier attributed as a subspecies of the Daubenton's bat Myotis daubentonii s. lato, is given. Fourteen species of bloodsucking arthropods feed on M. petax; these species include 4, 3, and 7 species of gamasid mites, fleas, and bloodsucking flies, respectively.

  8. 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.

  9. 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.

  10. Primary embryonic cells of Rhipicephalus microplus and Amblyomma cajennense ticks as a substrate for the development of Borrelia burgdorferi (strain G39/40)

    OpenAIRE

    Rezende,J.; CP. Rangel; NC. Cunha; AH. Fonseca

    2012-01-01

    Borrelia burgdorferi, the agent of Lyme borreliosis, is a spirochetes transmitted by ticks to humans and animals. Its cultivation in vitro in tick cells allows studies of its biology and provides methodology for future research in Brazil, and for the isolation of Borrelia spp. We examined in vitro the characteristics of embryonic cells of Rhipicephalus microplus and Amblyomma cajennense in cell culture and investigated the suitability of embryonic cells as a substrate for cultivation of B. bu...

  11. 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.

  12. Population-based passive tick surveillance and detection of expanding foci of blacklegged ticks Ixodes scapularis and the Lyme disease agent Borrelia burgdorferi in Ontario, Canada.

    Science.gov (United States)

    Nelder, Mark P; Russell, Curtis; Lindsay, L Robbin; Dhar, Badal; Patel, Samir N; Johnson, Steven; Moore, Stephen; Kristjanson, Erik; Li, Ye; Ralevski, Filip

    2014-01-01

    We identified ticks submitted by the public from 2008 through 2012 in Ontario, Canada, and tested blacklegged ticks Ixodes scapularis for Borrelia burgdorferi and Anaplasma phagocytophilum. Among the 18 species of ticks identified, I. scapularis, Dermacentor variabilis, Ixodes cookei and Amblyomma americanum represented 98.1% of the 14,369 ticks submitted. Rates of blacklegged tick submission per 100,000 population were highest in Ontario's Eastern region; D. variabilis in Central West and Eastern regions; I. cookei in Eastern and South West regions; and A. americanum had a scattered distribution. Rates of blacklegged tick submission per 100,000 population were highest from children (0-9 years old) and older adults (55-74 years old). In two health units in the Eastern region (i.e., Leeds, Grenville & Lanark District and Kingston-Frontenac and Lennox & Addington), the rate of submission for engorged and B. burgdorferi-positive blacklegged ticks was 47× higher than the rest of Ontario. Rate of spread for blacklegged ticks was relatively faster and across a larger geographic area along the northern shore of Lake Ontario/St. Lawrence River, compared with slower spread from isolated populations along the northern shore of Lake Erie. The infection prevalence of B. burgdorferi in blacklegged ticks increased in Ontario over the study period from 8.4% in 2008 to 19.1% in 2012. The prevalence of B. burgdorferi-positive blacklegged ticks increased yearly during the surveillance period and, while increases were not uniform across all regions, increases were greatest in the Central West region, followed by Eastern and South West regions. The overall infection prevalence of A. phagocytophilum in blacklegged ticks was 0.3%. This study provides essential information on ticks of medical importance in Ontario, and identifies demographic and geographic areas for focused public education on the prevention of tick bites and tick-borne diseases.

  13. 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.

  14. 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.

  15. 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).

  16. Contribution of the Infection-Associated Complement Regulator-Acquiring Surface Protein 4 (ErpC to Complement Resistance of Borrelia burgdorferi

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    Claudia Hammerschmidt

    2012-01-01

    Full Text Available Borrelia burgdorferi evades complement-mediated killing by interacting with complement regulators through distinct complement regulator-acquiring surface proteins (CRASPs. Here, we extend our analyses to the contribution of CRASP-4 in mediating complement resistance of B. burgdorferi and its interaction with human complement regulators. CRASP-4 (also known as ErpC was immobilized onto magnetic beads and used to capture proteins from human serum. Following Western blotting, factor H (CFH, CFH-related protein 1 (CFHR1, CFHR2, and CFHR5 were identified as ligands of CRASP-4. To analyze the impact of native CRASP-4 on mediating survival of serum-sensitive cells in human serum, a B. garinii strain was generated that ectopically expresses CRASP-4. CRASP-4-producing bacteria bound CFHR1, CFHR2, and CFHR5 but not CFH. In addition, transformed spirochetes deposited significant amounts of lethal complement components on their surface and were susceptible to human serum, thus indicating that CRASP-4 plays a subordinate role in complement resistance of B. burgdorferi.

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

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    Ashley R Dresser

    2009-12-01

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

  18. Borreliose de Lyme simile: uma doença emergente e relevante para a dermatologia no Brasil Lyme borreliosis simile: an emergent and relevant disease to dermatology in Brazil

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    Adivaldo Henrique da Fonseca

    2005-04-01

    Full Text Available Neste trabalho de revisão são apresentadas doenças relacionadas com espiroquetas do gênero Borrelia, agentes etiológicos de diferentes enfermidades comuns ao homem e a animais. Enfatizou-se a Borrelia burgdorferi lato sensu, que inclui diferentes espécies causadoras de doenças e com envolvimento sistêmico, com interesse em várias especialidades médicas, como dermatologia, reumatologia, cardiologia e neurologia. Considerando que existem diferenças quanto ao agente etiológico, além dos aspectos clínicos e laboratoriais, quando comparada com a borreliose de Lyme causada pelas Borrelia burgdorferi, B. garinii e B. afzelli, a infecção no Brasil deve ser referida como borreliose de Lyme simile. O eritema migratório recidivante é a principal manifestação clínica da borreliose existente tanto no Brasil como nos demais países. Essa lesão clássica está relacionada com a picada do carrapato vetor e inicia-se como uma mácula ou pápula cutânea avermelhada, de caráter expansivo, eventualmente surgem lesões semelhantes múltiplas a distância. A manifestação clínica da enfermidade, em especial o envolvimento cutâneo, é o parâmetro diagnóstico mais relevante, e os exames complementares sorológicos confirmam a suspeita clínica.This review article presents diseases related to spirochetes of the genus Borrelia, which are the etiological agents of many human and animal diseases. Focus was given to the Borrelia burgdorferi sensu lato complex, including nine different species that cause diseases often with multisystemic involvement and raising interest to many medical specialties, such as Dermatology, Rheumatology, Cardiology and Neurology. Due to differences concerning the etiologic agent, clinical and laboratorial presentations, when comparing with Borrelia burgdorferi, B. garinii and B. afzelli, the infection must be referred as Lyme disease-like illness in Brazil. The recurrent erythema migrans is the main clinical

  19. 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.

  20. Pós-graduação stricto sensu em anestesiologia: experiência de dez anos na UNESP

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    José Reinaldo Cerqueira Braz

    2005-08-01

    Full Text Available JUSTIFICATIVA E OBJETIVOS: O número de Programas de Pós-Graduação em Anestesiologia stricto sensu existente no país ainda é muito pequeno. Com a finalidade de incentivar a pós-graduação em Anestesiologia no Brasil, é apresentada a experiência acumulada em dez anos de atividades do programa da Universidade Estadual Paulista (UNESP. CONTEÚDO: O Programa de Pós-Graduação em Anestesiologia stricto sensu da UNESP foi credenciado pela CAPES, desde a sua criação em 1994, nos Cursos de Mestrado e Doutorado. O Programa é desenvolvido em três Áreas de Concentração: Risco e Proteção de Órgãos e Sistemas em Anestesia e Cirurgia; Qualidade e Segurança em Anestesiologia; e Modelos Clínicos e Experimentais em Terapia Antálgica, com as suas respectivas linhas de pesquisa, em número de 14. O número de alunos regulares do Programa é compatível com o número de orientadores (12, com proporção média de três alunos por orientador. Desde o seu início até setembro de 2004, ocorreram 45 Dissertações de Mestrado e 24 de Doutorado, perfazendo 69 defesas, a maioria com bolsas e financiamentos de Órgãos de Fomento à Pesquisa. Após a conclusão do doutorado, 65% dos alunos têm atividade de docência e pesquisa em instituições públicas e privadas do ensino universitário do país. A maioria das publicações do programa tem sido realizada em revistas nacionais com Qualis A, com menor número de publicações em revistas internacionais Qualis A ou B. O programa recebeu da CAPES o conceito 4,0 numa escala de 1 a 7, em suas três últimas avaliações. CONCLUSÕES: O programa tem se desenvolvido muito bem nos 10 anos de sua existência, alcançando os principais objetivos, como a formação de professores e pesquisadores na área de Anestesiologia para as instituições universitárias do país.

  1. Efeito do fogo na produção de frutos de Qualea parviflora Mart. (Vochysiaceae em cerrado sensu stricto

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    Alexandre Cesar Palermo

    2012-08-01

    Full Text Available Entre os muitos efeitos causados por queimadas nas espécies arbóreas nativas do Cerrado, a alteração da produção de frutos tem destaque, pois implica diretamente na manutenção de populações locais. O objetivo deste trabalho foi investigar o efeito de uma queimada prescrita na produção de frutos de Qualea parviflora Mart. em área de Cerrado sensu stricto em Brasília, DF. Foram utilizadas duas áreas de Cerrado sensu stricto, uma submetida a regime de queima bienal em agosto (queimada e outra protegida contra a queima há 14 anos (controle. Em cada área, 10 indivíduos reprodutivos foram selecionados e a sua produção de botões florais, flores e frutos, acompanhada durante 22 meses. Dois meses após uma queimada prescrita em 26 de agosto de 2008, a produção de estruturas reprodutivas foi maior na área queimada do que na área-controle. Contudo, a proporção de botões que geram flores e frutos ("fruit set" foi a mesma nas duas áreas. Na estação reprodutiva seguinte, um ano após a queima não houve produção de inflorescências na área queimada. No entanto, na área-controle a produção de frutos foi maior do que no período anterior. A ausência de produção de flores nos indivíduos da área queimada um ano após o fogo sugere que Q. parviflora pode necessitar de mais de um ano para retornar ao seu ciclo normal de reprodução, indicando que os efeitos de médio e de longo prazo devem ser considerados ao se usar o fogo como instrumento de manejo para redução do combustível em áreas de Cerrado.

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

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    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.

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

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    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.

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

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    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.

  5. 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

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    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

  6. 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

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    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

  7. Monophyly and interrelationships of Snook and Barramundi (Centropomidae sensu Greenwood) and five new markers for fish phylogenetics.

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    Li, Chenhong; Ricardo, Betancur-R; Smith, Wm Leo; Ortí, Guillermo

    2011-09-01

    Centropomidae as defined by Greenwood (1976) is composed of three genera: Centropomus, Lates, and Psammoperca. But composition and monophyly of this family have been challenged in subsequent morphological studies. In some classifications, Ambassis, Siniperca and Glaucosoma were added to the Centropomidae. In other studies, Lates+Psammoperca were excluded, restricting the family to Centropomus. Recent analyses of DNA sequences did not solve the controversy, mainly due to limited taxonomic or character sampling. The present study is based on DNA sequence data from thirteen genes (one mitochondrial and twelve nuclear markers) for 57 taxa, representative of all relevant species. Five of the nuclear markers are new for fish phylogenetic studies. The monophyly of Centropomidae sensu Greenwood was supported by both maximum likelihood and Bayesian analyses of a concatenated data set (12,888 bp aligned). No support was found for previous morphological hypotheses suggesting that ambassids are closely allied to the Centropomidae. Putative affinities between centropomids and Glaucosoma, Niphon, or Siniperca were strongly rejected by topology tests. In agreement with previous molecular hypotheses, our results place Centropomidae within a group of fishes that includes carangoids (e.g., jacks, remoras, dolphinfish, roosterfish, and cobia), flatfishes, barracudas, archerfishes, billfishes, moonfish, and threadfins. The phylogeny for the extant Centropomidae is ((Lates, Psammoperca), Centropomus).

  8. Behavioral Finance: an investigation based on the prospect theory and the investor profile of portuguese stricto sensu graduate students

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    Fabricio Luis Colognese Haubert

    2014-04-01

    Full Text Available Behavioral Finance is a new area of study that seeks to analyze the psychological aspects of individuals in making financial decisions. Studies in this area are intended to verify assumptions made by more traditional theories, in particular, the rationality of economic agents, and they aim to improve the economic and financial model through the incorporation of evidence on investor irrationality. In this context, the objective of this article is to understand the behavior of portuguese post-graduate students to its operations in investments based on behavioral finance. In order to learn about the behavioral profile of investors and prospective investors, a survey was conducted to collect quantitative data using a questionnaire comprising 15 questions for 130 stricto sensu graduate students of Lisbon – Portugal. It was possible to identify the predominant conservative and moderate profiles. It was observed that students showed risk aversion in the field of gains and risk propensity in the field of losses. So it is possible to say that the reflection effect occurred in this study.

  9. 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.

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    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

  10. Freqüência de anticorpos homólogos anti-Borrelia burgdorferi em eqüinos na mesorregião metropolitana de Belém, Estado do Pará Occurrence of homologous anti-Borrelia burgdorferi antibodies in equines in the metropolitan mesorregion of Belém, State of Para, Brazil

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    Katiany R. Galo

    2009-03-01

    Full Text Available Espiroquetas transmitidas por carrapatos são microrganismos de ampla distribuição geográfica e acometem animais silvestres, domésticos e seres humanos. Procedeu-se a análise sorológica de 300 soros de eqüinos onde 58 animais eram do município Ananideua, 61 eram de Belém, 131 de Castanhal e 50 eram do município de Santa Izabel do Pará para Borrelia burgdorferi através do teste ELISA indireto. Não foram observadas diferenças significativas (P Spirochaetes transmitted by ticks are microorganisms of worldwide distribution, which infect wild, domestic animals and human beings. A total of 300 equine sera from four municipalities: Ananideua (58, Belém (61, Castanhal (131, and Santa Izabel do Pará (50, were evaluated for Borrelia burgdorferi by an Elisa test. There were no significant differences (P<0.05 among municipalities, breed, sex or husbandry. A total of 80 (26.7% horses were B. burgdorferi positive with titles of 1:800, 72 (90% horses, 1:1.600, 6 (7.5% horses, and 1:3.200, 2 (2.5% horses. The results were similar to those in the USA, where related frequencies ranged from 7 to 75% in asymptomatic seropositive horses. The presence of anti-B.burgdorferi homologous antibodies in horses from four municipalities in the metropolitan mesorregion of Belém suggests the possibility of occurrence of human cases in the region.

  11. Cirurgias êntero-colorretais: abordagem cirúrgica de 129 pacientes do SUS no Programa de Pós-Graduação Sensu Lato em coloproctologia Abdominal entero-colorectal surgery: surgical approach of 129 patients from a public health service from a Post Graduate Program (Residency in coloproctology

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    Rodrigo Guimarães Oliveira

    2010-09-01

    Full Text Available Dentro do Programa de pós-graduação em Coloproctologia, durante o ano 2009, os dois pós-graduandos de segundo ano realizaram, como cirurgiões principais, 129 cirurgias de grande porte, sempre assistidos, efetivamente, por um ou dois preceptores. Todas as cirurgias foram realizadas em pacientes do SUS, na Santa Casa de Belo Horizonte, com absoluta presença dos membros do Grupo de Coloproctologia da Santa Casa e Faculdade de Ciências Médicas de Minas Gerais (GCP-SCBH-FCMMG. Foi feita uma análise retrospectiva dos 129 prontuários, permitindo várias observações importantes. A média etária dos pacientes foi 56,9 anos, com extremos de 25 e 87 anos, sendo as sexta e sétima décadas a mais representativa, respectivamente com 25,6% e 24,8%, totalizando 50,4% dos 55 pacientes (p0,05. A entidade nosológica mais comum foi o câncer colorretal (74 casos; 57,4%, seguindo as ileostomias (16 casos; 12,4% e as complicações cirúrgicas (11 casos; 8,5%. As cirurgias mais realizadas foram as retossigmoidectomia com anastomose colorretal (35 casos; 27,1%, as hemicolectomias direitas com anastomose ileo-transverso (20 casos; 15,5% e o restabelecimento de trânsito intestinal de ileostomia (16 casos;12,4%. Das 129 cirurgias 53 (41,1% não envolveram anastomoses e 76 (58,9% envolveram ressecções intestinais e anastomoses. Houve oito co-morbidades (6,2%, sendo a caquexia (três casos a mais comum. Houve 17 complicações (13,2%, 11 envolvendo as 76 ressecções com anastomose (14,5% e seis as ressecções sem anastomoses (11,3%. As complicações mais comuns entre as 11 provenientes de ressecções e anastomoses foram as deiscências (sete; 9,2%. As anastomoses mecânicas (55 complicaram mais (16,3% que as manuais (21 (9,5%. Houve 14 óbitos (10,8%, sendo seis (4,6% devidos à sepse, quatro (3,1% devido a TEP e quatro (3,1% devido a falência múltipla de órgãos. Dos 14 óbitos, quatro (3,1% foram decorrentes de complicações cirúrgicas e dez (7,7% foram decorrentes de co-morbidades.In the framework of postgraduate Coloproctology for the year 2009, the two graduate students conducted the second year as principal surgeons, 129 major surgeries, always assisted effectively by one or two tutors. All surgeries were performed on public patients in Santa Casa de Belo Horizonte, with absolute presence of members of the Coloproctology Unit of Santa Casa School of Medical Sciences of Minas Gerais (GCP-CBHS-FCMMG. A retrospective analysis of 129 medical records was carried out, allowing several important observations. The average age of patients was 56.9 years, with extremes of 25 and 87 years, while the sixth and seventh decades the most representative, with respectively 25.6% and 24.8% to 50.4% of 55 patients (p 0.05. The most common nosological entity was colorectal cancer (74 cases, 57.4%, following the ileostomies (16 cases, 12.4% and surgical complications of previous surgeries (11 cases, 8.5%. The most commonly performed procedures were abdominal rectosigmoidectomy with colorectal anastomosis (35 cases, 27.1%, the right hemicolectomy with ileo-transverse anastomosis (20 cases, 15.5% and the resumption of intestinal transit of ileostomy (16 cases, 12, 4%. Of 129 surgeries 53 (41.1% did not involve anastomosis and 76 (58.9% involved intestinal resection and anastomosis. There were eight co-morbidities (6.2% and cachexia (three cases the most common. There were 17 complications (13.2%, 11 involving the 76 resections with anastomosis (14.5% and six resections without anastomosis (11.3%. The most common complications among the 11 patients from resection and anastomosis were dehiscence (seven, 9.2%. The mechanical anastomosis (55 developed more complications (16.3% than handmade anastomosis (21 (9.5%. There were 14 deaths (10.8%, six (4.6% due to sepsis, four (3.1% due to pulmonary thromboembolic disease and four (3.1% due to multiple organ failure. Of the 14 deaths, four (3.1% were due to surgical complications and ten (7.7% were due to co-morbidities.

  12. Cirurgia no câncer colorretal: abordagem cirúrgica de 74 pacientes do SUS portadores de câncer colorretal em programa de pós-graduação lato sensu em coloproctologia Surgery in colorectal cancer: surgical approach of 74 patients from the Brazilian National Health System with colorectal cancer in a postgraduate program (residency in coloproctology

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    Rodrigo Guimarães Oliveira

    2011-03-01

    Full Text Available A análise retrospectiva de 74 prontuários de pacientes do Sistema Único de Saúde (SUS, operados de câncer colorretal pelo Residente R2 supervisionado e auxiliado por preceptores, permitiu as seguintes conclusões: a média etária dos pacientes foi 57,2, sendo as sexta e sétima décadas responsáveis por 51,4% dos pacientes. O câncer retal foi preponderante nas mulheres (54,1%. As localizações mais comuns dos tumores foram no sigmoide (31,1%, reto alto (24,3% e ceco (17,6%. As cirurgias mais realizadas foram a retossigmoidectomia com anastomose colorretal (36,6%, e hemicolectomia direita com anastomose íleo-transverso (21,7%. As características anatômicas dos tumores, baseadas na classificação TNM, mais comuns foram: T3 (62,1%, N0 (59,5% e M0 (77,0% (pIn the framework of postgraduate Coloproctology for 2009, two graduate students conducted the second year as principal surgeons, 129 major surgeries, always assisted effectively by one or two tutors. All surgeries were performed on public patients in Santa Casa de Belo Horizonte, with absolute presence of members of the Grupo de Coloproctologia da Santa Casa de Belo Horizonte e Faculdade de Ciências Médicas de Minas Gerais (GCP-CBHS-FCMMG. A retrospective analysis of 74 medical records of patients from the Brazilian National Health System, resected of colorectal cancer by Resident R2, supervised and assisted by mentors, could get into the following conclusions: the average age of patients was 57.2, and the sixth and seventh decades accounted for 51.4% of the patients. The rectal cancer was predominant in women (54.1%. The most common sites of tumors were in the sigmoid (31.1%, rectum (24.3%, and cecum (17.6%. The most commonly performed procedures were retossigmoidectomy with colorectal anastomosis (36.6% and right hemicolectomy with ileo-transverse anastomosis (21.7%. The anatomical characteristics of the tumors based on TNM classification findings were: T3 (62.1%, N0 (59.5%, and M0 (77.0% (p<0.05. The average number of lymph nodes found in surgical specimens was 10.4. Sixty-three anastomoses (85.1% were carried out, being 38 (60.3% mechanical and 25, manual (39.7%. There were 14 comorbidities (18.9%, the cachexia having more prominence (eight cases. The rate of surgical complications was 12.2% (nine cases, and the surgeries that have caused more complications were total colectomy with ileo-rectal anastomosis (40.0%, and double stapled abdominal retossigmoidectomy (20%. The most common complications were anastomotic fistula (five cases. Complications (nine were more caused by comorbidities (seven than by the surgical procedure (two. The surgeries that required less time were: laparotomy with ileostomy (average of 75 minutes and with colostomy (average of 95 minutes, and the longest times were occupied by total proctocolectomy with definite ileostomy (240 minutes and left hemicolectomy with transverse-rectal anastomosis (240 minutes, with an average length of 160 minutes. The smaller specimens were those resulting from Hartmann's procedure (29 cm and retossigmoidectomy (32 cm; and the most extensive specimens were of total colectomy with ileorectal anastomosis (120 cm and total proctocolectomy with terminal ileostomy (150 cm, with the mean at 34.5. There were 12 deaths (16.2%, two of which directly related to surgery and the other related to clinical complications and comorbities

  13. Biodiversité, bioactivité et biosynthèse des composés terpéniques volatils des lavandes ailées, Lavandula stoechas sensu lato, un complexe d'espèces méditerranéennes d'intérêt pharmacologique

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    Benabdelkader, Tarek

    2012-01-01

    In this work, we have assessed the composition and the biological activities of essential oils (EO) extracted from L. stoechas collected from 11 locations in northern Algeria. The oils were analyzed by GC/FID and GC/MS. A total of 121 compounds were identified that accounted for 69.88-91.2 % of the total oil contents. Major components were fenchone (11.27-37.48 %), camphor (1.94-21.8 %), 1,8-cineol (0.16-8.71 %) and viridiflorol (2.89-7.38 %). In vitro biological activities demonstrated that ...

  14. 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

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    Lin, Tao; 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 wer...

  15. 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.

  16. 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

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    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

  17. 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.

    2016-01-01

    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

  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. Flora arbórea da bacia do rio Tibagi (Paraná, Brasil: Celastrales sensu Cronquist Tree flora of the Tibagi river basin (Paraná, Brazil: Celastrales sensu Cronquist

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    Ricardo Augusto Gorne Viani

    2007-06-01

    Full Text Available Este trabalho estudou, por meio de coleções de herbários, os representantes arbóreos da ordem Celastrales sensu Cronquist, encontrados na bacia do rio Tibagi, estado do Paraná, Brasil. Esta bacia hidrográfica, subdividida em três zonas de norte para sul, baixo Tibagi (BT, médio Tibagi (MT e alto Tibagi (AT, apresenta diferentes condições ambientais e tipos de vegetação ao longo de sua extensão. A ordem Celastrales está representada na bacia estudada por 15 espécies arbóreas, pertencentes às famílias Aquifoliaceae, Celastraceae e Icacinaceae. Icacinaceae conta com apenas duas espécies, Citronella gongonha e C. paniculata, sendo a primeira distinta pelo ovário glabro e folhas geralmente com espinhos. Aquifoliaceae contém seis espécies: Ilex brasiliensis, I. brevicuspis, I. chamaedryfolia, I. dumosa, I. paraguariensis e I. theezans, que ocorrem preferencialmente no AT e MT e se distinguem pela dimensão, revestimento, ápice e margem da folha e pela morfologia das sépalas. Celastraceae está representada por sete espécies pertencentes a dois gêneros, Plenckia populnea, espécie de cerrado, encontrada apenas no MT e seis espécies de Maytenus (M. aquifolia, M. dasyclada, M. evonymoides, M. ilicifolia, M. robusta e M. salicifolia, com distinção baseada principalmente no tipo de margem e dimensão das folhas, forma do ramo e número de flores por inflorescência.A study of the tree species of the order Celastrales sensu Cronquist from the Tibagi river basin, Paraná state, Brazil, is presented, based on herbarium material. This basin is subdivided into three zones, from north to south: lower Tibagi (BT, mid Tibagi (MT and upper Tibagi (AT, each with different environmental conditions and vegetation types. The order Celastrales is represented in the basin by 15 tree species belonging to three families: Aquifoliaceae, Celastraceae and Icacinaceae. Icacinaceae has only two species, Citronella gongonha and C. paniculata. The

  20. Investigating the potential role of non-vls genes on linear plasmid 28–1 in virulence and persistence by Borrelia burgdorferi

    OpenAIRE

    Magunda, Petronella R. Hove; Bankhead, Troy

    2016-01-01

    Background The lp28-1 plasmid is required for persistent infection by the Lyme disease spirochete, Borrelia burgdorferi. Mutational studies on this plasmid have shown that the vls locus is important for antigenic variation of the VlsE lipoprotein that leads to immune evasion and persistence. However, it is still unknown whether the vls system is the only genetic locus on this plasmid necessary for long-term infection, and thus the potential role of non-vls genes on lp28-1 in virulence and per...

  1. 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.

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

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    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.

  3. Estrutura de um cerrado strico sensu na Gleba Cerrado Pé-de-Gigante, Santa Rita do Passa Quatro, SP

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    Fidelis Alessandra Tomaselli

    2003-01-01

    Full Text Available O Cerrado ocupa aproximadamente 23% do território brasileiro e 70% do bioma correspondem a cerrado stricto sensu (s.s., sendo relevantes os estudos que buscam o entendimento da estrutura da vegetação nessas áreas. Com esse objetivo, foram estudadas cinco parcelas (10×25m em um hectare de cerrado s.s., amostrando-se todos os indivíduos com perímetro no nível do solo igual ou acima de 3cm (exceto lianas e indivíduos mortos. Alguns parâmetros fitossociológicos foram analisados, assim como a distribuição de classes de diâmetro dos indivíduos amostrados e a estrutura vertical. Foram amostradas 1.747 indivíduos, distribuídos em 75 espécies, pertencentes a 31 famílias. A densidade total absoluta encontrada foi de 13.976 ind.ha-1 e a área basal total, de 4,902m². Leguminosae foi a família com o maior número de espécies (16. As espécies que apresentaram os maiores valores de Índice do Valor de Importância (IVI foram Anadenanthera falcata (Benth. Altschul, Myrcia guianensis (Aubl. DC., Xylopia aromatica (Lm. Mart., Ouratea spectabilis (Mart. Engl. e Pouteria ramiflora (Mart. Radlk. O Índice de Shannon encontrado foi de 3,623. A distribuição de classes de diâmetro apresentou curva na forma de "J" invertido, estando a maioria dos indivíduos na primeira classe. A área estudada não apresentou estratos bem definidos, estando a maioria dos indivíduos entre 1 e 3m de altura.

  4. The mode of action of spatial repellents and their impact on vectorial capacity of Anopheles gambiae sensu stricto.

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    Sheila B Ogoma

    Full Text Available Malaria vector control relies on toxicity of insecticides used in long lasting insecticide treated nets and indoor residual spraying. This is despite evidence that sub-lethal insecticides reduce human-vector contact and malaria transmission. The impact of sub-lethal insecticides on host seeking and blood feeding of mosquitoes was measured. Taxis boxes distinguished between repellency and attraction inhibition of mosquitoes by measuring response of mosquitoes towards or away from Transfluthrin coils and humans. Protective effective distance of coils and long-term effects on blood feeding were measured in the semi-field tunnel and in a Peet Grady chamber. Laboratory reared pyrethroid susceptible Anopheles gambiae sensu stricto mosquitoes were used. In the taxis boxes, a higher proportion of mosquitoes (67%-82% were activated and flew towards the human in the presence of Transfluthrin coils. Coils did not hinder attraction of mosquitoes to the human. In the semi-field Tunnel, coils placed 0.3 m from the human reduced feeding by 86% (95% CI [0.66; 0.95] when used as a "bubble" compared to 65% (95% CI [0.51; 0.76] when used as a "point source". Mosquitoes exposed to coils inside a Peet Grady chamber were delayed from feeding normally for 12 hours but there was no effect on free flying and caged mosquitoes exposed in the semi-field tunnel. These findings indicate that airborne pyrethroids minimize human-vector contact through reduced and delayed blood feeding. This information is useful for the development of target product profiles of spatial repellent products that can be used to complement mainstream malaria vector control tools.

  5. Molecular evidence for bicontinental hybridogenous genomic constitution in Lepidium sensu stricto (Brassicaceae) species from Australia and New Zealand.

    Science.gov (United States)

    Mummenhoff, Klaus; Linder, Peter; Friesen, Nikolai; Bowman, John L; Lee, Ji-Young; Franzke, Andreas

    2004-02-01

    Lepidium sensu stricto (s.s.) (Brassicaceae) (ca. 150 species) is distributed worldwide with endemic species on every continent. It is represented in Australia and New Zealand by 19 and seven native species, respectively. In the present study we used a nuclear ribosomal internal transcribed spacer (ITS) phylogeny in comparison with a cpDNA phylogeny to unravel the origin of Australian/New Zealand species. Although phylogenetic relationships within Lepidium s.s. were not fully resolved, the cpDNA data were in agreement with a Californian origin of Lepidium species from Australia/New Zealand. Strongly conflicting signals between the cp- and nuclear DNA phylogenetic analysis clearly indicated hybridogenous genomic constitution of Australian Lepidium s.s. species: All 18 studied Australian/New Zealand Lepidium s.s. species examined shared a Californian cpDNA type. While eleven Australian/New Zealand species appeared to harbor a Californian ITS type, a group of seven species shared a South African ITS type. This pattern is most likely explained by two trans-oceanic dispersals of Lepidium from California and Africa to Australia/New Zealand and subsequent hybridization followed by homogenization of the ribosomal DNA either to the Californian or South African ITS type in the two different lineages. Calibration of our molecular trees indicates a Pliocene/Pleistocene origin of Lepidium in Australia/New Zealand. Low levels of cpDNA and ITS sequence divergence and unresolved topologies within Australian/New Zealand species suggest a rapid and recent radiation of Lepidium after the hybridization event. This coincides with dramatic climatic changes in that geological epoch shaping the composition of the vegetation.

  6. Sporothrix schenckii (sensu strict S. globosa) mating type 1-2 (MAT1-2) gene.

    Science.gov (United States)

    Kano, Rui; Anzawa, Kazushi; Mochizuki, Takashi; Nishimoto, Katsutaro; Hiruma, Masataro; Kamata, Hiroshi; Hasegawa, Atsuhiko

    2013-09-01

    Sporotix schenckii is a pathogenic fungus that causes human and animal sporotrichosis, and based on morphology of the sessile conidia and molecular analysis, it was recently recognized as a species complex comprising at least the following six sibling species: S. albicans, S. brasiliensis, S. globosa, S. luriei, S. mexicana and S. schenckii. However, apart from S. schenckii sensu strict, only S. brasiliensis, S. globosa and S. luriei are associated with human and animal infection. S. globosa has been most commonly isolated in Asia, Europe and the USA; therefore, molecular epidemiological study for S. globosa is important in relation to human sporotrichosis in Japan. To the best of our knowledge, this is the first study to determine the mating type 1-2 (MAT1-2) gene of Sporothrix schenckii with the aim of understanding the taxonomy of the genus Sporothrix. The MAT1-2 gene (1618 bp) encodes a protein sequence of 198 amino acids. Reverse transcription polymerase chain reaction analysis also detected MAT1-2 gene mRNA expression in all of the S. schenckii strains examined, indicating that this gene is expressed in S. schenckii cells. Phylogenetic analysis of the MAT1-2 gene fragments of Ophiostoma himal-ulmi, O. novo-ulmi, O. ulmi and S. schenckii indicated that these isolates could be classified into four clusters. MAT1-1 gene-specific polymerase chain reaction was positive in 15 isolates, but negative in four human isolates and one feline isolate.

  7. The Mode of Action of Spatial Repellents and Their Impact on Vectorial Capacity of Anopheles gambiae sensu stricto

    Science.gov (United States)

    Ogoma, Sheila B.; Ngonyani, Hassan; Simfukwe, Emmanuel T.; Mseka, Antony; Moore, Jason; Maia, Marta F.; Moore, Sarah J.; Lorenz, Lena M.

    2014-01-01

    Malaria vector control relies on toxicity of insecticides used in long lasting insecticide treated nets and indoor residual spraying. This is despite evidence that sub–lethal insecticides reduce human–vector contact and malaria transmission. The impact of sub–lethal insecticides on host seeking and blood feeding of mosquitoes was measured. Taxis boxes distinguished between repellency and attraction inhibition of mosquitoes by measuring response of mosquitoes towards or away from Transfluthrin coils and humans. Protective effective distance of coils and long-term effects on blood feeding were measured in the semi–field tunnel and in a Peet Grady chamber. Laboratory reared pyrethroid susceptible Anopheles gambiae sensu stricto mosquitoes were used. In the taxis boxes, a higher proportion of mosquitoes (67%–82%) were activated and flew towards the human in the presence of Transfluthrin coils. Coils did not hinder attraction of mosquitoes to the human. In the semi–field Tunnel, coils placed 0.3 m from the human reduced feeding by 86% (95% CI [0.66; 0.95]) when used as a “bubble” compared to 65% (95% CI [0.51; 0.76]) when used as a “point source”. Mosquitoes exposed to coils inside a Peet Grady chamber were delayed from feeding normally for 12 hours but there was no effect on free flying and caged mosquitoes exposed in the semi–field tunnel. These findings indicate that airborne pyrethroids minimize human–vector contact through reduced and delayed blood feeding. This information is useful for the development of target product profiles of spatial repellent products that can be used to complement mainstream malaria vector control tools. PMID:25485850

  8. 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