Pesquisa de anticorpos anti-Borrelia burgdorferi em cães da região de Cotia, São Paulo, Brasil

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.Com a finalidade de estudar a ocorrência da doença de Lyme em cães no Brasil, o teste imunoenzimático (ELISA) indireto, utilizado para o diagnóstico da doença no homem, foi padronizado para a espécie canina. Utilizou-se como antígeno a cepa americana de Borrelia burgdorferi G 39/40. Os soros de cães procedentes da região de Cotia, área de risco para ocorrência da doença de Lyme, foram testados e os resultados positivos foram confirmados através da técnica de "Western blotting". Para investigação de possíveis reações cruzadas, os soros foram também testados para diferentes variantes sorológicas de Leptospira interrogans e L. biflexa pela técnica de soroaglutinação microscópica. Dos 237 soros testados pela técnica de ELISA, 23 (9,7%) foram positivos, sendo que 20 (86,9%) destes soros foram também positivos pela técnica de "Western blotting", sugerindo que a região de Cotia seja área de risco para doença de Lyme. Embora 4 (1,7%) dos soros testados apresentassem positividade para doença de Lyme e para leptospirose concomitantemente, não foi observada associação entre estes resultados (X² = 0,725 e p = 0,394), sugerindo ausência de reatividade sorológica cruzada entre as duas enfermidades

Detection of anti-Borrelia and anti-Babesia antibodies in the serum of children with clinical manifestations and compatible epidemiology with Lyme-Like disease in the State of Mato Grosso do Sul

A ocorrência de manifestações clínicas e laboratoriais semelhantes às encontradas na doença de Lyme e da coinfecção com a babesiose já foi demonstrada em trabalhos anteriores em adultos, porém não existem estudos desta natureza em crianças. OBJETIVO: Caracterizar o perfil clínico-epidemiológico da síndrome de Lyme-Símile em crianças do Estado de Mato Grosso do Sul e avaliar a prevalência dos anticorpos anti-Borrelia burgdorferi e anti-Babesia bovis no soro de pacientes que preencheram os critérios estabelecidos. MÉTODOS: 100 pacientes entre 9 meses e 16 anos de idade foram submetidos à pesquisa dos anticorpos pela técnica de ELISA e a soroprevalência foi comparada com um grupo-controle. RESULTADOS: Positividade para anticorpos anti-B. burgdorferi ocorreu em 27% dos pacientes com suspeita clínica, sendo 17% IgM e 12% IgG. As manifestações articulares ocorreram em 21 pacientes, as manifestações cutâneas em três pacientes e as manifestações neurológicas em três pacientes. A prevalência de anticorpos contra B. burgdorferi nos pacientes do grupo-controle foi de 15% (p < 0,05). Os anticorpos anti-B. bovis estiveram presentes em 24% dos pacientes suspeitos e em 3% do grupo-controle (p < 0,05). Concomitância dos anticorpos ocorreu em 10 pacientes com suspeita clínica e em nenhum controle. CONCLUSÃO: Os resultados deste estudo evidenciam a etiologia de caráter infeccioso e reacional das manifestações clínicas encontradas nos pacientes, podendo caracterizar uma síndrome, que possivelmente ocorra na presença de múltiplos microrganismos e está relacionada com carrapatos. Foi encontrada freqüência superior de anticorpos contra Borrelia e Babesia nos pacientes em relação aos controles. Embora pequena, essa diferença foi estatisticamente significante. Este achado não atesta que o quadro clínico dos pacientes seja causado por espécies de Borrelia ou Babesia, mas levantam a possibilidade de participação de algum agente infeccioso que possa estar relacionado a esses microrganismos. Os estudos devem prosseguir para melhorar a delineação desta síndrome em nosso meio, assim como isolar o agente etiológico.The occurence of clinical and laboratory manifestations similar to the Lyme disease and the coinfection with babesiosis was already demonstraded in previous researches with adult patients. However, there are no studies in children. OBJECTIVE: To identify the clinical-epidemiological profile of the Lyme Simile Syndrome in children of the State of Mato Grosso do Sul and to evaluate the prevalence of antibodies against Borrelia burgdorferi and Babesia bovis in the serum of patients that had fulfilled the established criteria. METHODS: One hundred patients (age range: 9 months and 16 years-old) were screened for the presence of those antibodies, using ELISA, comparing to a control group. RESULTS: Antibodies against B. burgdoferi were found in 27% of the patients with a clinical picture. The articular manifestations occurred in 21 patients, cutaneous in 3 and neurological in 3. The prevalency in the control group was 15% (p < 0.05). The antibodies for Babesia bovis were present in 24% of suspect patients and in 3% of the control group (p < 0.05). Concomitance of the two antibodies occurred in 10 patients with clinical suspect and no one of control group. CONCLUSION: The results found in this study suggest the infective and reactive character of the clinical manifestations of the patients, which can characterize a Syndrome, which possibly occurs in the presence of multiple microorganisms that can be associated with tick-borne. It was found a higher frequency of antibodies against borrelia and babesia in patients in relation to the controls. Although small, this difference was statistically significant. This finding does not certify that the clinical features of patients is caused by species of borrelia or babesia, but raised the possibility of involvement of some infectious agent that could be related to these micro-organisms. The studies should continue in order to improve the delineation of this syndrome and to try the isolation of the etiologic agent in our State.Fundação de Apoio à Pesquisa (FAEPA

Report on the unusual presence of latent microorganisms in animals: a risk to research and health of employees?

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.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.Universidade de São Paulo Faculdade de MedicinaUSP Faculdade de Medicina Veterinária e Zootecnia Departamento de Medicina Veterinária e Saúde AnimalUniversidade Federal de São Paulo (UNIFESP)FMUSP Hospital das ClínicasFMUSPFMUSP Hospital das Clínicas Laboratório de Investigação Médica em ReumatologiaUNIFESPSciEL

Histomorphometric analysis of cutaneous remodeling in the early stage of the scleroderma model

INTRODUCTION/OBJECTIVES: Systemic sclerosis, or scleroderma, is a rheumatic disease characterized by autoimmunity, vasculopathy, and fibrosis of the skin and several internal organs. In the present study, our aim was to assess the skin alterations in animals with scleroderma during the first stages of disease induction. METHODS: To induce scleroderma, female New Zealand rabbits (n = 12) were subcutaneously immunized with 1 mg/ml of collagen V (Col V) in complete Freund's adjuvant, twice with a thirty-day interval. Fifteen days later, the animals received an intramuscular booster with type V collagen in incomplete Freund's adjuvant, twice with a fifteen-day interval. The control group was inoculated with 1 ml of 10 mM acetic acid solution diluted with an equal amount of Freund's adjuvant. Serial dorsal skin biopsies were performed at 7, 15, and 30 days and stained with H&E, Masson's trichrome and Picrosírius for morphological and morphometric analyses. RESULTS: Immunized rabbits presented a significant increase in collagen in skin collected seven days after the first immunization (p=0.05). CONCLUSION: The results from this experimental model may be very important to a better understanding of the pathogenic mechanisms involved in the beginning of human SSc. Therapeutic protocols to avoid early remodeling of the skin may lead to promising treatments for SSc in the future

Changes in histoanatomical distribution of types I, III and V collagen promote adaptative remodeling in posterior tibial tendon rupture

INTRODUCTION: Posterior tibial tendon dysfunction is a common cause of adult flat foot deformity, and its etiology is unknown. PURPOSE: In this study, we characterized the morphologic pattern and distribution of types I, III and V collagen in posterior tibial tendon dysfunction. METHOD: Tendon samples from patients with and without posterior tibial tendon dysfunction were stained by immunofluorescence using antibodies against types I, III and V collagen. RESULTS: Control samples showed that type V deposited near the vessels only, while surgically obtained specimens displayed type V collagen surrounding other types of collagen fibers in thicker adventitial layers. Type III collagen levels were also increased in pathological specimens. On the other hand, amounts of collagen type I, which represents 95% of the total collagen amount in normal tendon, were decreased in pathological specimens. CONCLUSION: Fibrillogenesis in posterior tibial tendon dysfunction is altered due to higher expression of types III and V collagen and a decreased amount of collagen type I, which renders the originating fibrils structurally less resistant to mechanical forces

Changes in Histoanatomical Distribution Of Types I, III And V Collagen Promote Adaptative Remodeling in Posterior Tibial Tendon Rupture

INTRODUCTION: Posterior tibial tendon dysfunction is a common cause of adult flat foot deformity, and its etiology is unknown. PURPOSE: In this study, we characterized the morphologic pattern and distribution of types I, III and V collagen in posterior tibial tendon dysfunction. METHOD: Tendon samples from patients with and without posterior tibial tendon dysfunction were stained by immunofluorescence using antibodies against types I, III and V collagen. RESULTS: Control samples showed that type V deposited near the vessels only, while surgically obtained specimens displayed type V collagen surrounding other types of collagen fibers in thicker adventitial layers. Type III collagen levels were also increased in pathological specimens. On the other hand, amounts of collagen type I, which represents 95% of the total collagen amount in normal tendon, were decreased in pathological specimens. CONCLUSION: Fibrillogenesis in posterior tibial tendon dysfunction is altered due to higher expression of types III and V collagen and a decreased amount of collagen type I, which renders the originating fibrils structurally less resistant to mechanical forces

Identificação sorológica de Rickettsia spp do grupo da febre maculosa em capivaras na região de Campinas, SP, Brasil

Diseases transmitted by ticks have been an important health problem all over the world. Brazilian Spotted Fever (BSF) stands for a serious epidemiological concern due to the high mortality rates pointed out. Capybaras are commonly incriminated as possible reservoirs in the BSF transmission cycle. In the last decades the numbers of these animals raised sharply and they have invaded human areas. They intensify the contact between ticks and humans beings. This study aim is to contribute to the possible role performed for this rodent in the BSF epidemiology in some areas located in Campinas region, São Paulo. Cabybaras infected by rickettsiae of BSF group were studied through the analysis of the frequencies of BSF-group rickettisae antibodies titer = 64 by indirect immunofluorescence test (IFA), and data from human cases epidemiological surveillance. The serum frequency positiveness varied greatly according to areas where animals were captured. However it was found serum positiviness only in the areas where human cases of BSF were reported. These findings suggest the capybara may be seen as sentinel animal. Due to presence of serological cross reactivity between microorganisms belonging to SF group, the results must be interpreted carefully and additional methods to distinguish pathogenic rickettsiae are required in our country38616941699Doenças transmitidas por carrapatos vêm sendo um importante problema de saúde pública no mundo. A Febre Maculosa Brasileira (FMB) representa um sério risco epidemiológico devido às altas taxas de letalidade apresentadas. As capivaras são freqüentemente incriminadas como possíveis reservatórios no ciclo de transmissão da FMB. Nas últimas décadas o número desses animais cresceu intensamente e eles invadiram os espaços humanos. As capivaras intensificam o contato entre carrapatos e seres humanos na medida em que se apresentam muito infestadas por estes parasitos. O objetivo deste estudo é contribuir para o conhecimento do possível papel desempenhado por este roedor na epidemiologia da FMB em algumas áreas da região de Campinas, SP. Foi estudada a infecção das capivaras por rickettsias do grupo da FMB, por meio da análise das freqüências de anticorpos contra este grupo, nestes animais, e dados da vigilância epidemiológica de casos humanos. A freqüência desses anticorpos variou amplamente entre as localidades, entretanto, só foram encontrados soros com anticorpos com titulagem =64 naquelas onde havia notificação de casos humanos. Estes achados sugerem que a capivara poderá ser um animal sentinela. No entanto, devido à ocorrência de reação cruzada entre os microorganismos do grupo de FM estes resultados devem ser interpretados com cautela e são necessários métodos capazes de distinguir rickettsias patogênica

Identificação sorológica de Rickettsia spp do grupo da febre maculosa em capivaras na região de Campinas, SP, Brasil

Diseases transmitted by ticks have been an important health problem all over the world. Brazilian Spotted Fever (BSF) stands for a serious epidemiological concern due to the high mortality rates pointed out. Capybaras are commonly incriminated as possible reservoirs in the BSF transmission cycle. In the last decades the numbers of these animals raised sharply and they have invaded human areas. They intensify the contact between ticks and humans beings. This study aim is to contribute to the possible role performed for this rodent in the BSF epidemiology in some areas located in Campinas region, São Paulo. Cabybaras infected by rickettsiae of BSF group were studied through the analysis of the frequencies of BSF-group rickettisae antibodies titer = 64 by indirect immunofluorescence test (IFA), and data from human cases epidemiological surveillance. The serum frequency positiveness varied greatly according to areas where animals were captured. However it was found serum positiviness only in the areas where human cases of BSF were reported. These findings suggest the capybara may be seen as sentinel animal. Due to presence of serological cross reactivity between microorganisms belonging to SF group, the results must be interpreted carefully and additional methods to distinguish pathogenic rickettsiae are required in our country.Doenças transmitidas por carrapatos vêm sendo um importante problema de saúde pública no mundo. A Febre Maculosa Brasileira (FMB) representa um sério risco epidemiológico devido às altas taxas de letalidade apresentadas. As capivaras são freqüentemente incriminadas como possíveis reservatórios no ciclo de transmissão da FMB. Nas últimas décadas o número desses animais cresceu intensamente e eles invadiram os espaços humanos. As capivaras intensificam o contato entre carrapatos e seres humanos na medida em que se apresentam muito infestadas por estes parasitos. O objetivo deste estudo é contribuir para o conhecimento do possível papel desempenhado por este roedor na epidemiologia da FMB em algumas áreas da região de Campinas, SP. Foi estudada a infecção das capivaras por rickettsias do grupo da FMB, por meio da análise das freqüências de anticorpos contra este grupo, nestes animais, e dados da vigilância epidemiológica de casos humanos. A freqüência desses anticorpos variou amplamente entre as localidades, entretanto, só foram encontrados soros com anticorpos com titulagem =64 naquelas onde havia notificação de casos humanos. Estes achados sugerem que a capivara poderá ser um animal sentinela. No entanto, devido à ocorrência de reação cruzada entre os microorganismos do grupo de FM estes resultados devem ser interpretados com cautela e são necessários métodos capazes de distinguir rickettsias patogênicas