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

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

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

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.

Lyme disease in Haryana, India.

Lyme disease is a multiorgan animal‑borne disease caused by the spirochete Borrelia burgdorferi. This case series highlights its presence in Haryana, a nonendemic zone. The first case was a 27‑year‑old housewife who presented with an annular erythematous patch with a central papule following an insect bite on the left upper arm. The second case was a 32‑year‑old farmer who gave a history of insect bite on the right arm followed by the development of an erythematous patch with a central blister. The third case, a 17‑year‑old boy presented with a history of tick bite over right thigh and a typical bull’s eye lesion with central ulceration. These cases were managed with oral doxycycline 100 mg twice daily for 14 days. The fourth case was a 7‑year‑old boy with typical erythema migrans on the right check and neck while the fifth case, a 30‑year‑old housewife, presented with an erythematous patch with a central papule on the right buttock. These patients were treated with oral amoxycillin 25 mg/kg, thrice daily for 14 days. All patients showed IgM antibodies to B. burgdorferi. Treatment led to clearance of lesions in all the patients. Lyme borreliosis was diagnosed in these patients based on the history of established exposure to tick bites, presence of classic signs and symptoms, serology and the response to treatment.

Evidence of Borrelia in wild and domestic mammals from the state of Minas Gerais, Brazil / Evidência de Borrelia em mamíferos silvestres e domésticos no Estado de Minas Gerais, Brasil

The main of the study was to evaluate the presence of Borrelia burgdorferi infection in domestic and wild vertebrates and ectoparasites in endemic areas from the state of Minas Gerais, Brazil. A total of 445 serum samples were examined by ELISA, which used the Borrelia burgdorferi strain G39/40 U.S. source and 3,821 tick samples were tested by polymerase chain reaction (PCR). B. burgdorferi antibodies were found in 30 serum samples (6.74%); three in marsupials (7.69%), three in rodents (2.80%), nine in dogs (6.25%), and 15 in horses (9.68%). Nested-PCR performed in DNA samples obtained from collected ticks demonstrated negative results. Although attempts to amplify B. burgdorferi DNA from ticks had been not successful, the presence of seroreactive vertebrates suggests the possibility the Borrelia species circulating in these regions. Further research is required to provide information on the presence of Borrelia in Brazilian territory and its association with Baggio-Yoshinari syndrome.

O principal objetivo do estudo foi avaliar a presença de infecção por Borrelia burgdorferi em vertebrados domésticos e silvestres e ectoparasitas em áreas endêmicas do estado de Minas Gerais, Brasil. Um total de 445 amostras de soro foram examinadas por ELISA, onde usou-se a cepa americana G39/40 de Borrelia burgdorferi e 3.821 amostras de carrapatos foram testados pela reação em cadeia da polimerase (PCR). Anticorpos anti -B. burgdorferi foram encontrados em 30 amostras de soro (6,74%); três marsupiais (7,69%), três em roedores (2,80%), em nove cães (6,25%) e 15 em cavalos (9,68%). Nested-PCR realizada em amostras de DNA obtidas a partir de carrapatos coletados demonstraram resultados negativos. Apesar das tentativas para amplificar o DNA de B. burgdorferi a partir de carrapatos não tenha sido bem sucedido, a presença de soroatividade em vertebrados sugere a possibilidade de espécies de Borrelia circulando nestas regiões. Mais pesquisas são necessárias para fornecer informações sobre a presença de Borrelia em território brasileiro e sua associação com a Síndrome de Baggio-Yoshinari.

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

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.

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

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 < 0,05) entre os municípios, nem quanto à raça, sexo e função dos animais. Um total de 80 (26,7 por cento) animais foram positivos para B. burgdorferi com os títulos de 1:800, 72 (90 por cento) eqüinos; 1:1.600, 6 (7,5 por cento) eqüinos; e 1:3.200, 2 (2,5 por cento) eqüinos. Os resultados observados foram similares aos descritos nos EUA, onde foram relatadas freqüências de soropositivos variando entre 7 e 75 por cento em eqüinos assintomáticos. A presença de anticorpos homólogos contra B. burgdorferi em eqüinos na mesorregião metropolitana de Belém é indicativo da ampla distribuição do agente e da possibilidade de ocorrerem casos humanos deste agente na região.

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 percent) horses were B. burgdorferi positive with titles of 1:800, 72 (90 percent) horses, 1:1.600, 6 (7.5 percent) horses, and 1:3.200, 2 (2.5 percent) horses. The results were similar to those in the USA, where related frequencies ranged from 7 to 75 percent 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.

Description of Lyme disease-like syndrome in Brazil: is it a new tick borne disease or Lyme disease variation?

An emerging clinical entity that reproduces clinical manifestations similar to those observed in Lyme disease (LD) has been recently under discussion in Brazil. Due to etiological and laboratory particularities it is named LD-like syndrome or LD imitator syndrome. The condition is considered to be a zoonosis transmitted by ticks of the genus Amblyomma, possibly caused by interaction of multiple fastidious microorganisms originating a protean clinical picture, including neurological, osteoarticular and erythema migrans-like lesions. When peripheral blood of patients with LD-like syndrome is viewed under a dark-field microscope, mobile uncultivable spirochete-like bacteria are observed. PCR carried out with specific or conservative primers to recognize Borrelia burgdorferi sensu stricto or the genus Borrelia has been negative in ticks and in biological samples. Two different procedures, respectively involving hematoxylin and eosin staining of cerebrospinal fluid and electron microscopy analysis of blood, have revealed spirochetes not belonging to the genera Borrelia, Leptospira or Treponema. Surprisingly, co-infection with microorganisms resembling Mycoplasma and Chlamydia was observed on one occasion by electron microscopy analysis. We discuss here the possible existence of a new tick-borne disease in Brazil imitating LD, except for a higher frequency of recurrence episodes observed along prolonged clinical follow-up.

Anticorpos para Borrelia burgdorferi em indivíduos que trabalham com animais silvestres / Antibodies specific to Borrelia burgdorferi in wild animal workers

The occurrence of serum antibodies to Borrelia burgdorferi in professionals in close contact with wild animals was determined. Seventy eight technicians workers coming from two São Paulo public institutions housing wild animals had their blood collected (serum samples). All samples were submitted to ELISA for IgM and IgG antibodies against Borrelia burgdorferi. The results showed five positive (6.4 percent), two suspect (2.6 percent) and 71 negatives (91 percent) samples. Based on positive results it is concluded that the infection level is higher to that detected in the general population and similar to values of endemic areas, concluding that this assessed population could be considered at risk for Lyme disease.

Búsqueda de la espiroqueta borrelia burgdorferi sensu lato mediante PCR en garrapatas ixoideas chilenas silvestres / Search for the spirochete borrelia burgdorferi senso lato by polymerase chain reaction in Chilean ticks

Background: Lyme disease is a tick-borne human disease caused by the spirochete Borrelia burgdorferi. Main vectors of Lyme disease are ticks of the Ixodes and Amblyomma genera. Cases with clinical manifestations of Lyme disease and favorable responses to antimicrobial agents have been reported in Chile, some of them with erythema migrans, the hallmark of B burgdorferi infection. Aim: To detect the presence of B burgdorferi in Chilean ticks. Material and methods : A total of 62 ticks were recollected from wild rodents and cervidae in the Southern region of Chile. Infected and non infected ticks of the species Ixodes ricimus, were used as controls. Insects were homogenised and B burgdorferi was detected using classical and nested polymerase chain reactions. Results : B burgdorferi was not detected in the studied ticks. Conclusions: Although all the elements required for the enzootic cycle of B burgdorferi are present in Chile, its direct detection in Chilean ticks using the nested polymerase chain reaction assay was negative

Perfil da borreliose de Lyme no Brasil / Outline of Lyme borreliosis in Brazil

A partir de 1989, identificamos 25 pacientes com doenca de Lyme no Brasil, dos quais 15 no estagio agudo e dez na fase latente, que permitiram reforcar sua existencia, pelos seguintes motivos: 1) existencia de um perfil clinico, caracterizado por manifestacoes cutanea, gerais, articulares e raramente cardiacas; 2) demonstracao sorologica de anticorpos anti-Borrelia burgdorferi; 3) modificacao sorologica evolutiva dos casos agudos; 4) resposta boa aos antibioticos no estagio agudo e apenas satisfatoria dos casos latentes; 5) distribuicao geografica da casuistica permitiu o mapeamento das areas de risco; 6) identificacao de carrapatos Ixodideos; 7) visualizacao de microorganismos a microscopia de campo escuro, que lembram borrelias, em culturas de humanos, animais silvestres e carrapatos; 8) comprovacao da existencia de areas endemicas para borreliose de Lyme pela sorologia em caninos e bovinos

Neuroborreliosis en Chile: caso pediátrico de probable adquisición por mascotas importadas / Neuroborreliosis in Chile: pediatric case probably due to imported pets

Lyme disease, caused by spirochete Borrelia burgdorferi, has several clinical manifestations and is transmitted to man by tick bites. In Chile and Latin America, several cases have been reported, but none with inmunoblot confirmation or isolation of the infecting organism. We report a 9 year old boy consulting with bilateral facila palsy, polyradiculoneuritis with tetraparesis and meningeal irritation. Cerebrospinal fluis analysis showed increased protein concentration without pleocytosis and negative viral or bacterial cultures. IgM antibodies against Borrelia burgdorferi, were positive by ELISA and were confirmed by immunoblot at the Reference Laboratory of the University of Connecticut. The child had a recent contact with hamsters brought from Germany. The substantiation of Lyme disease existence in Chile should prompt the search and isolation of the causal agent

Contribuicao ao conhecimento da borreliose de Lyme no Brasil / Contribution to the knowledge of Lyme borreliosis in Brazil

Entre 1989 a 1995 identificamos 25 casos de borreliose de Lyme confirmados pela sorologia no Brasil. Trata-se de zoonose causada pela espiroqueta Borrelia burgdorferi e transmitida pela picada de carrapatos Ixodideos. Pacientes contrairam a enfermidade visitando litoral ou zonas rurais, e aproximadamente 50 por cento dos casos tiveram contacto com carrapatos. O aspecto distintivo da enfermidade e a presenca do eritema migratorio. A borreliose no Brasil pode cursar de diferentes modos, podendo simular quadro gripal, doencas reumatologicas como artralgias, artrites ou fadiga cronica e queixas neurologicas como neurites cranianas ou perifericas e encefalite (agitacao, depressao, disturbios de sono e comportamento)

Multiple sclerosis and positive lyme serology

As manifestaçöes neurológicas da doença de Lyme säo extremamente variadas e podem ocorrer de forma episódica e focal, às vezes simulando o quadro clínico da esclerose múltipla (EM). A situaçäo pode se tornar ainda mais confusa quando pacientes com diagnóstico de EM apresentam anticorpos séricos, anti-Borrelia burgdorferi, em áreas näo endêmicas para a doença de Lyme. Relatamos o caso de uma paciente de 45 anos de idade com o diagnóstico de EM devido a episódio de neurite óptica há 15 anos e desde entäo surtos de distúrbios motores e sensitivos no membro superior direito e nos membros inferiores, associados a disfunçäo esfinctérica. A reaçäo de ELISA para doença de Lyme foi positiva em dois laboratórios diferentes e a positividade foi confirmada por Western blot. Tratamento com ceftriaxone näo produziu mudança significativa do quadro clínico. Embora a paciente deva ter sido infectada pela espiroqueta, desenvolvendo entäo anticorpos séricos específicos, a relaçäo entre esta infecçäo pela Borrelia burgodorferi e a sintomatologia neurológica, no presente caso, permanece näo esclarecida

Enfermedad de lyme y su espectro expandido

La enfermedad de Lyme se puede manifestar por compromiso de diferentes órganos incluyendo la piel, el sistema nervioso central y el sistema musculoesquelético. Reportamos los casos de seis niños entre los seis y los 15 años de edad, vistos en una única institución,quienes ilustran las variadas manifestaciones de la enfermedad en sus fases aguda y crónica y las dificultades en su diagnóstico diferencial. En cuatro casos, la entidad fue adquirida en el Reino Unido, y en dos en países diferentes el diagnóstico y tratamiento de los pacientes con una forma de presentación aguda y sintomatología típica es relativamente fácil, pero la presencia de manifestaciones atípicas y curso crónico de la enfermedad en niños puede presentar retos considerables al clínico

La enfermedad de lyme

La enfermedad de Lyme es una borreliosis transmitida por garrapatas del género ixodes que parece estar limitada a las zonas templadas, especialmente del hemisferio norte. En Norteamérica y Europa ha adquirido gran importancia clínica y epidemiológica desde 1975. Se caracteriza clínicamente por tres etapas sucesivas. En la temprana localización,se presenta eritema alrededor del sitio de picadura de la garrapata y puede haber síntomas constitucionales. En la temprana diseminada hay lesiones dermatológicas, síntomas constitucionales severos y manifestaciones neurológicas,articulares o cardíacas. En la etapa tardía hay alteraciones crónicas en articulaciones, piel o sistema nervioso. Usualmente los pacientes no presentan ni todas las etapas ni todos los síntomas. El diagnóstico se base más a menudo en hallazgos clínicos que en métodos de laboratorio, debido a la falta de reproducibilidad de estos últimos. En las etapas iniciales la enfermedad de Lyme responde bien a antibióticos: tetraciclinas (especialmente doxiciclina) y penicilinas. Puesto que actualmente no hay vacuna, la prevención consiste en evitar contacto con plantas que tengan ixodes, y en remover tales artrópodos de la ropa o la piel, lo más pronto posible

Alternative culture medium for isolation and growth of Borrelia burgdorferi

Borrelia burgdofer é uma spirochete que é agente causador de doenças infecciosas: Doença Lyme (LD). Muitos meios de cultura säo descritos para isolamento e crescimento desse agente. O autor descreve um meio alternativo de cultivar em laboratório o vírus do Lyme spirochetes

Estudo epidemiologico da doenca de Lyme no Brasil / Epidemiological study of Lyme's disease in Brazil

A doenca de Lyme, uma espiroquetose transmitida por carrapatos, foi inicialmente relatada nos Estados Unidos da America em 1977, por Allen C. Steere, que descreveu a doenca no estado de Conecticut. Posteriormente, seguiu-se relatos da doenca nao somente em outros estados americanos, bem como em outros paises e continentes, como Canada, Uniao Sovietica, Japao, China, Australia e no Leste Europeu. Com o intuito de se realizar a pesquisa desta doenca no Brasil criou-se uma equipe multidisciplinar para realizacao deste projeto com a participacao de microbiologistas, entomologistas e clinicos da area reumatologica, pertencentes ao quadro da Universidade de Sao Paulo. O presente trabalho tem por objetivo descrever a elaboracao e a evolucao e uma infraestrutura para esta pesquisa, incluindo montagem de laboratorios para pesquisa imunologica, sorologica e cultura para borrelias, cooperacao com diferentes instituicoes nacionais de saude, elaboracao de pesquisa de campo, bem como mostrar os resultados preliminares encontrados por esta equipe.