Abstract Objective: To evaluate the frequency of anti-collagen type V in humans with early systemic sclerosis (SSc) compared to defined SSc patients and healthy controls, since collagen type V was shown to be overexpressed in early SSc patients’ skin and there is no data concerning the presence of this antibody in early stages of human SSc. Experimental studies showed that animal models immunized with collagen type V developed a disease similar to human systemic sclerosis (SSc), with antibodies production, mainly in early stages post-immunization. Methods: Eighty-one female SSc patients were included and divided into two groups: early-SSc (18 patients-EULAR Preliminary Criteria) and defined-SSc (63 patients-ACR Criteria 1980). The control group consisted of 19 healthy women age-matched to Early-SSc group. Anti-collagen type V was performed by ELISA. Data was analyzed by appropriate tests. Results: The prevalence of anti-collagen type V in early-SSc, defined-SSc and control groups was respectively 33, 17 and 5% (p = 0.07). SSc patients with anti-collagen type V had shorter disease duration compared to those without this antibody (8.8 ± 5.1 vs. 14.7 ± 8.9, p = 0.006). Likewise, early-SSc patients with anti-collagen V also had a shorter disease duration than patients negative for this antibody (4.6 ± 2.2 vs. 9.7 ± 5.2, p = 0.04). No association with clinical subsets or scleroderma antibodies specificities was observed (p > 0.05). Conclusion: The production of anti-collagen type V in SSc seems to be an early event independent of other antibodies specificities. Further studies are necessary to determine if the underlying mechanism for this chronology involves a primary immune response to abnormal expression of collagen type V.

Abstract Borreliosis caused by Borrelia burgdorferi sensu lato is a cosmopolitan zoonosis studied worldwide; it is called Lyme disease in many countries of the Northern Hemisphere and Lyme-like or Baggio-Yoshinari Syndrome in Brazil. However, despite the increasing number of suspect cases, this disease is still neglected in Brazil by the medical and veterinary communities. Brazilian Lyme-like borreliosis likely involves capybaras as reservoirs and Amblyomma and Rhipicephalus ticks as vectors. Thus, domestic animals can serve as key carriers in pathogen dissemination. This zoonosis has been little studied in horses in Brazil. The first survey was performed in the state of Rio de Janeiro, and this Brazilian Borreliosis exhibits many differences from the disease widely described in the Northern Hemisphere. The etiological agent shows different morphological and genetic characteristics, the disease has a higher recurrence rate after treatment with antibiotics, and the pathogen stimulates intense symptoms such as a broader immune response in humans. Additionally, the Brazilian zoonosis is not transmitted by the Ixodes ricinus complex. With respect to clinical manifestations, Baggio-Yoshinari Syndrome has been reported to cause neurological, cardiac, ophthalmic, muscle, and joint alterations in humans. These symptoms can possibly occur in horses. Here, we present a current panel of studies involving the disease in humans and equines, particularly in Brazil.

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.

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.

INTRODUÇÃO: Os sintomas da borreliose brasileira se assemelham às manifestações clínicas da Doença de Lyme (DL), porém, existem diferenças epidemiológicas e laboratoriais entre essas enfermidades. Primers normalmente utilizados para diagnosticar a DL não conseguiram detectar cepas de borrelia no Brasil. OBJETIVO: O objetivo desse trabalho foi identificar a borrelia brasileira usando um gene conservado que sintetiza o gancho flagelar (flgE) da Borrelia burgdorferi sensu lato. MÉTODO: Três pacientes com eritema migratório e epidemiologia positiva foram recrutados para o estudo. Amostras de sangue foram coletadas, e o DNA foi extraído por kits comerciais. RESULTADOS: O gene flgE foi amplificado a partir do DNA de todos os pacientes selecionados. Após o sequenciamento, essas amostras positivas revelaram homologia de 99% para B. burgdorferi. CONCLUSÃO: Estes resultados reforçam a existência de borreliose no Brasil. No entanto, não está claro se esta borreliose é causada por uma variante geneticamente modificada da B. burgdorferi sensu stricto ou por uma nova espécie de Borrelia spp.

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.

O objetivo do estudo foi avaliar o perfil epidemiológico, clínico e laboratorial dos pacientes com Síndrome Baggio-Yoshinari (SBY), internados no Instituto de Infectologia Emilio Ribas, São Paulo, Brasil, no período de julho de 1990 a julho de 2006. SBY é uma nova doença transmitida por carrapatos, causada pela Borrelia burgdorferi sensu lato, que se assemelha a Doença de Lyme (DL), exceto pelas particularidades epidemiológicas, clínicas e laboratoriais. A partir dos registros de 60 pacientes com sorologia positiva para B. burgdorferi pelos métodos de ELISA e Western-blotting, 19 casos foram diagnosticados como SBY, de acordo com critérios adotados pelo LIM-17 HCFMUSP, laboratório de referência para a pesquisa de SBY no Brasil. Os outros 41 pacientes restantes foram classificados como tendo infecções diversas ou processos auto-imunes. O início dos sintomas no grupo SBY variou de um dia a seis anos. Quatro dos 19 pacientes foram incluídos na fase aguda da doença e 15 na fase latente. Os sintomas gerais inespecíficos foram identificados em quase todos os casos, com altas freqüências de febre (78,9%) e linfoadenomegalia (36,8%). Seis pacientes tiveram lesões de pele (31,5%); seis artralgia ou artrite (31,5%) e oito sintomas neurológicos (42%). Curiosamente, dois pacientes apresentaram anticorpos para B. burgdorferi exclusivamente no líquido cefalorraquidiano. Uma vez que a SBY é uma zoonose emergente brasileira e seu diagnóstico é complexo, todo o conhecimento novo sobre SBY deve ser difundido para os médicos brasileiros, com o objetivo de ensiná-los a diagnosticar esta surpreendente doença transmitida por carrapatos, evitando-se a sua progressão para sequelas crônicas irreversíveis.

The aim of this study was to evaluate the epidemiological, clinical and laboratorial profile of patients with Baggio-Yoshinari Syndrome (BYS), who underwent internment at the Instituto de Infectologia Emilio Ribas in São Paulo, Brazil, during the period from July 1990 to July 2006. BYS is a new Brazilian tick-borne disease caused by Borrelia burgdorferi sensu lato microorganisms that resembles features of Lyme disease (LD), except for its epidemiological, clinical and laboratorial particularities. From 60 patients' records with positive serology to B. burgdorferi done by ELISA and Western-blotting methods, 19 cases were diagnosed as having BYS, according to criteria adopted at LIM-17 HCFMUSP, the Brazilian Reference Laboratory for the research of BYS. The other 41 remaining patients displayed miscellaneous infections or auto-immune processes. The beginning of symptoms in BYS group varied from one day to six years, from the onset of the disease. Four of 19 patients were included in acute disease stage, and 15 in latent. General unspecific symptoms were identified in almost all cases, with high frequencies of fever (78.9%) and lymphadenomegaly (36.8%). Six patients had skin lesions (31.5%); six arthralgia or arthritis (31.5%) and eight neurological symptoms (42%). Interestingly, two patients showed antibodies directed to B. burgdorferi exclusively in cerebrospinal fluid. Since BYS is a new emergent Brazilian zoonosis and its diagnosis is sometimes complex, all the new knowledge about BYS must be scattered to Brazilian Medical specialists, aiming to teach them how to diagnose this amazing tick-borne disease and to avoid its progression to chronic irreversible sequels

The aim of the present paper was to evaluate cyst formation and growth parameters of Borrelia garinii in a range of media differing in formulation and cost. A qualitative assessment of morphology and motility of B. garinii was conducted. All media were prepared aseptically and used in test tubes or Petri dishes. For each medium, the initial spirochete concentration was standardized to 10³ spirochets/mL. The following culture media were suitable to grow B. garinii: Barbour-Stoenner-Kelly, brain heart infusion and PMR. Growth was minimal at six weeks post-inoculation and maximum spirochete density was observed between 9-12 weeks. Often, the cultures developed cysts of different sizes, isolated or in groups, with a spiraled portion of variable sizes, mainly in unfavorable culture media. Brazilian Lyme disease-like illness, also known as Baggio-Yoshinari syndrome (BYS), is a new and interesting emerging tick-borne disease, caused by Borrelia burgdorferi sensu lato spirochetes, only during its cystic forms. It has been assumed that the peculiar clinical and laboratory features of BYS are consequential to the absence of a human sucker Ixodes ricinus complex tick at risk areas in Brazil, supporting the concept that the borrelia phenotypic expression pattern is modified as it is transmitted through the host.

A Doença de Lyme (DL) é uma zoonose frequente no hemisfério Norte e considerada uma enfermidade infecciosa causada por espiroquetas do complexo Borrelia burgdorferi sensu lato e transmitida pela picada de carrapatos do grupo Ixodes ricinus. Os primeiros casos semelhantes à DL no Brasil foram descobertos, em 1992, em irmãos que após serem picados por carrapatos desenvolveram eritema migratório, sintomas gripais e artrite. Criteriosa análise da casuística brasileira, mostrou que os aspectos epidemiológicos, clínicos e laboratoriais no país, divergiam bastante dos exibidos pelos pacientes com DL nos Estados Unidos da América e Eurásia. Não foram encontrados carrapatos do complexo Ixodes ricinus hematófago ao homem nas áreas de risco; a enfermidade no país é recorrente; a Borrelia burgdorferi jamais foi isolada no Brasil e os ensaios sorológicos específicos exibem positividade baixa e oscilante. Além disso, o exame do sangue periférico dos pacientes quando analisados à microscopia eletrônica exibe estruturas sugestivas de Mycoplasma spp, Chlamydia spp e bacteroides. Na verdade, estas estruturas podem representar as formas latentes das espiroquetas (forma L ou bactérias desprovidas de parede), adaptadas a sobreviver em condições inóspitas em hospedeiros vertebrados e invertebrados. Assim, a zoonose presente no país recebeu a denominação de Síndrome Baggio-Yoshinari e definida como: "Enfermidade infecciosa nova e emergente brasileira, transmitida por carrapatos não pertencentes ao complexo Ixodes ricinus, causada por espiroquetas na sua morfologia atípica e latente, que origina manifestações clínicas semelhantes às observadas na DL, exceto pela ocorrência de recidivas clínicas e desordens autoimunes".

Lyme disease (LD) is a frequent zoonosis found in the Northern Hemisphere and is considered an infectious disease caused by spirochetes belonging sensu lato to the Borrelia burgdorferi complex transmitted by ticks of the Ixodes ricinus group. In 1992, first cases similar to LD were described in Brazil, when brothers, after a tick bite episode developed symptoms , as erythema migrans, general flu-like symptoms and arthritis. Careful analysis of Brazilian LD-like illness casuistry showed that epidemiological, clinical and laboratorial features in the country were very different from those exhibited by North American and Eurasian LD patients. Human blood-suckers Ixodes ricinus complex ticks were absent at risk areas; the disease is recurrent in the country; Borrelia burgdorferi was never isolated in Brazil and specific serologic tests have shown little positivity with inconsistent results. Furthermore, peripheral blood analysis of patients on electron microscopy exhibited structures resembling Mycoplasma spp, Chlamydia spp and spirochete-like microorganisms. In fact, they were assumed to be latent forms of spirochetes (L form or cell wall deficient bacteria) adapted to survive at inhospitable conditions in vertebrate and invertebrate hosts. For these reasons, the Brazilian zoonosis was named Baggio-Yoshinari Syndrome (BYS) and defined as: "Exotic and emerging Brazilian infectious disease, transmitted by ticks not belonging to the Ixodes ricinus complex, caused by latent spirochetes with atypical morphology, which originates LD-like symptoms, except for occurrence of relapsing episodes and auto-immune disorders".

Relata-se inusitado achado de micro-organismos, semelhantes às espiroquetas, móveis e de diferentes morfologias e tamanhos, identificados pela microscopia de campo escuro, no sangue de animais do Centro de Bioterismo da Faculdade de Medicina da USP. As bactérias não crescem em meios habituais de cultivo, são pouco coradas pelo Giemsa e derivados da prata, e as provas sorológicas e moleculares foram negativas para Borrelia e Leptospira. A microscopia eletrônica revelou presença de micro-organismos com morfologia de Mycoplasma e, devido à sua mobilidade, sugeriu-se a presença de Mollicutes do gênero Spiroplasma. Visualizaram-se micro-organismos com mesmas características em 15 dos 26 funcionários (57,6%) do Centro de Bioterismo da FMUSP; contudo, exames clínicos e laboratoriais indicaram que os indivíduos estavam saudáveis. Estudos adicionais desenvolvidos na Disciplina de Reumatologia da FMUSP mostraram que, aproximadamente, 94% dos pacientes com Síndrome Baggio-Yoshinari (SBY) e 20% dos indivíduos normais também exibiam as mesmas estruturas identificadas no Centro de Bioterismo. A microscopia eletrônica realizada com amostras de pacientes com SBY mostrou bactérias que apresentam semelhanças com Mycoplasma, Chlamydia e Bacteroides. Visto que as sorologias e os testes moleculares foram negativos para estes contaminantes e com base em publicações da literatura médica, sugeriu-se que estes agentes infecciosos latentes fossem bactérias na sua apresentação L, que são definidas como bactérias despojadas de parede celular, assumindo, assim, morfologia de Mycoplasma, sendo, na maioria dos casos, inofensivas aos hospedeiros. Concluímos que os "espiroquetídeos" visualizados em animais e funcionários do Centro de Bioterismo seriam bactérias na forma L, não patogênicas, provenientes de contaminantes do meio ambiente, infecções regulares habituais ou oriundos de micro-organismos endógenos da flora saprofítica normal. Ao contrário, os "espiroquetídeos" identificados na SBY, ao preservarem a capacidade de invadir células in vitro, são potencialmente patogênicos e relacionados com a etiologia da SBY. Interpretamos a SBY como zoonose nova e particular ao território brasileiro, causada por espiroquetas adaptadas às formas latentes, possivelmente provenientes de mutações bacterianas em resposta às condições ecológicas e geográficas próprias do país.

We report the unusual finding of mobile spirochetal microorganisms with different morphologies and sizes, on dark-field microscopy of the blood of animals from the Vivarium of the Medical School of USP. The bacteria did not grow in common culture media, shows faint staining to Giemsa and silver-derived stains, and serologies and molecular tests were negative for Borrelia and Leptospira. Electron microscopy revealed the presence of microorganisms with Mycoplasma-like morphology and, due to its mobility, it was suggested that they represented Mollicutes of the genus Spiroplasma. Microorganisms with the same morphology were also observed in 15 out of 26 employees (57.6%) of the Vivarium of FMUSP; however, clinical and laboratorial exams indicated that those individuals were healthy. Additional studies undertaken at the Rheumatology Department of FMUSP demonstrated the presence of the same structures identified at the Vivarium in approximately 94% of the patients with Baggio-Yoshinary syndrome (BYS) and 20% of healthy individuals. Electron microscopy of the blood of BYS patients showed bacteria that shared similarities with Mycoplasma, Chlamydia, and Bacteroides. Since serologies and molecular tests were negative for those contaminants, and based on publications in the medical literature, it was suggested that those latent infectious agents were L-form bacteria, defined as cell wall deficient bacteria, assuming, therefore, Mycoplasma morphology and they are, for the most part, harmless to the host. We concluded that spirochetal microorganisms visualized in animals and employees of the Vivarium were non-pathogenic L-form bacteria from contaminants in the environment, regular infections, or endogenous microorganism from the normal saprophytic flora. On the other hand, spirochetal organisms identified in BYS, by preserving the capacity to invade cells in vitro, are potentially pathogenic and related to the etiology of BYS. We consider BYS as a novel Brazilian zoonosis caused by spirochetes adapted to their latent form, possibly due to bacterial mutations in response to ecologic and geographic conditions unique to Brazil.

INTRODUÇÃO: A doença de Lyme (DL) é uma doença de picada de carrapato, causado pela espiroqueta Borrelia burgdorferi sensu lato, transmitida por carrapatos do complexo Ixodes ricinus, que promove múltiplas manifestações clínicas sistêmicas. No Brasil, uma síndrome diferente é descrita e mimetiza sintomas de DL, mas também se manifesta com alta frequência de episódios recorrentes e manifestações alérgicas e imunológicas. É transmitida pelo carrapato Amblyomma cajennense e o agente etiológico é uma espiroqueta não cultivável de forma atípica. Devido a essas particularidades, esta zoonose emergente tem sido denominada síndrome brasileira semelhante à doença de Lyme ou síndrome de Baggio-Yoshinari (SBY). OBJETIVO: Descrever o espectro da manifestação neurológica da SBY. PACIENTES: Foram analisados 30 pacientes com SBY e sintomas neurológicos. RESULTADOS: A média de idade dos pacientes foi de 34,2 ± 13,3 anos (6 a 63 anos); 20 eram mulheres e 10 homens. Um alto número de episódios recorrentes (73,6%) e distúrbios psiquiátricos e psicossociais graves (20%) foram características típicas. Eritema migrans similar ao visto em hemisfério norte foi identificado em 43,3% dos pacientes no início da doença. A recorrência das lesões cutâneas diminuiu com a progressão da doença. Sintomas articulares (artrite) aconteceram em aproximadamente metade dos pacientes com SBY no início e durante o episódio de recidiva. CONCLUSÕES: A SBY é considerada uma nova doença transmitida por carrapato no Brasil que difere da clássica DL observada no hemisfério norte. A SBY reproduz sintomas neurológicos observados na DL, exceto pela presença adicional de recorrência de episódios e uma tendência de causar manifestações neurológicas crônicas e articulares.

INTRODUCTION: Lyme disease (LD) is a tick-borne disease, caused by Borrelia burgdorferi sensu lato spirochetes, transmitted by Ixodes ricinus complex ticks, which leads to multiple systemic clinical manifestations. In Brazil, a different syndrome is described that mimics LD symptoms, but that also manifests high frequencies of recurrent episodes and immune-allergic manifestations. It is transmitted by the Amblyomma cajennense tick and the etiological agent is an uncultivable spirochete with atypical morphology. Due to its particularities, this emerging zoonosis has been called Brazilian LD-like syndrome or Baggio-Yoshinari Syndrome (BYS). OBJECTIVE: To describe the neurological spectrum of BYS. PATIENTS: Thirty patients with neurological symptoms of BYS were analyzed. RESULTS: Mean age of patients was 34.2 ± 13.3 years old (6 to 63 years); 20 were females and 10 males. A high number of recurrent episodes (73.6%) and severe psychiatric or psycho-social disturbances (20%) were distinguishing features. Erythema migrans similar to those seen in the Northern hemisphere was identified in 43.3% of patients at disease onset. The recurrence of skin lesions diminished as the disease progressed. Articular symptoms (arthritis) happened in nearly half of patients at BYS onset and during relapsing episodes. CONCLUSIONS: The BYS is considered a new tick borne disease in Brazil that differs from classical LD observed in the Northern hemisphere. BYS replicates most of the neurological symptoms observed in LD, except for the additional presence of relapsing episodes and the tendency to cause chronic neurological and articular manifestations.