First serological detection of Borrelia spp. in dogs in western Cuba / Primeira detecção sorológica de Borrelia spp. em cães na região oeste de Cuba

Abstract This study aimed to verify the presence of IgG antibodies against Borrelia burgdorferi sensu lato (s.l) in domestic dogs in western Cuba. Serum samples were analyzed by indirect enzyme-linked immunosorbent assay (ELISA), using crude antigens of a B. burgdorferi strain of North American origin. To verify the presence of Borrelia spp., deoxyribonucleic acid (DNA) extracted from individual blood samples was analyzed by nested-PCR, with markers targeted for amplification of portions of the flagellin B gene (flaB) present in Borrelia spirochetes. Ticks were also collected through inspection of the animals. Sera from 93 of 176 (52.84%) dogs were reactive to the indirect ELISA. Geographic prevalence varied from 54.35% (25/46) in Boyeros, 44.44% (20/45) in Cotorro, 66.67% (22/33) in Habana del Este, and 50% (26/52) in San José de las Lajas. There was no statistical difference between these tested variables. No blood samples analyzed were positive for the Borrelia flaB gene.

Resumo Este estudo teve como objetivo confirmar a presença de anticorpos IgG contra Borrelia burgdorferi sensu lato (s.l) em cães na região oeste de Cuba. As amostras de soro foram analisadas por ensaio de imunoabsorção enzimática (ELISA) indireto, usando-se antígenos brutos de uma cepa de B. burgdorferi de origem norte-americana. Para confirmar a presença de Borrelia spp., o ácido desoxirribonucleico (DNA), extraído de amostras individuais de sangue, foi analisado por PCR, utilizando-se marcadores direcionados para a amplificação de porções do gene da flagelina B (flaB) presente nas espiroquetas de Borrelia. Os carrapatos também foram coletados através da inspeção dos animais. Os soros de 93 de 176 (52,84%) cães foram reativos ao ELISA indireto. A prevalência geográfica variou de 54,35% (25/46) em Boyeros, 44,44% (20/45) em Cotorro, 66,67% (22/33) em Habana del Este e 50% (26/52) em San José de las Lajas. Não houve diferença estatística entre essas variáveis testadas. Nenhuma amostra de sangue analisada foi positiva para o gene Borrelia flaB.

Enfermedad de Lyme: a propósito de un caso clínico importado / Lyme disease: about an imported case report

Resumen Presentamos el caso clínico de una mujer norteamericana que consultó en nuestro país por un eritema migrans múltiple, diagnosticándose una enfermedad de Lyme. Este cuadro infeccioso es causado por espiroquetas del complejo Borrelia burgdorferi sensu lato (Bbsl) y es transmitido por la mordedura de garrapatas. Tradicionalmente Bbsl había sido detectada en garrapatas sólo en el hemisferio norte. Sin embargo, desde el 2013 ha habido reportes en Sudamérica. En Chile, recientemente se describió Borrelia chilensis, la cual no tiene una enfermedad asociada en humanos. Se discuten aspectos del agente infeccioso, su epidemiología, sus vectores y nuevos hallazgos en Sudamérica. Además, se plantean los criterios diagnósticos clínicos, de laboratorio y tratamiento, de acuerdo a la etapa en su historia natural.

This is a case report of an american woman who consulted in our country for multiple erythema migrans, from which a Lyme disease was diagnosed. This infectious disease is caused by spirochetes from the Borrelia burgdorferi sensu lato complex (Bbsl) and is transmitted by the bite of ticks. Traditionally Bbsl had been detected in ticks only in the Northern Hemisphere. However, since 2013 there have been reports in South America. In Chile, Borrelia chilensis was recently described, which does not have an associated disease in humans. Aspects of the infectious agent, its epidemiology, its vectors and new findings in South America are discussed. Likewise, the clinical diagnostic criteria, laboratory and appropriate treatment are proposed, according to the stage in their natural history.

Síndrome de Lyme crónico con manifestaciones neurológicas: a propósito de un caso / Chronic Lyme syndrome with neurological manifestations: A case report

RESUMEN El síndrome de Lyme crónico representa un término controversial que agrupa un conjunto de síntomas persistentes e inespecíficos, en su mayoría reumatológicos o neurológicos, dentro de los cuales se encuentra la enfermedad postLyme, cuya incidencia epidemiológica es baja, el conocimiento fisiopatológico es controversial y el abordaje diagnóstico se basa especialmente en la interpretación clínica, dirigiendo su enfoque terapéutico hacia la resolución sintomática. Se presenta un caso de una mujer de 17 años, con enfermedad de Lyme de reciente diagnóstico, de aparente evolución de 3 años, con diagnóstico previo de enfermedad desmielinizante, que consultó por deterioro neurológico. Se revisan consideraciones científicas sobre su aproximación clínica y principios terapéuticos, los cuales se contrastan con los aplicados.

SUMMARY Chronic Lyme syndrome represents a controversial term that groups together a set of persistent and nonspecific symptoms, mostly rheumatological and / or neurological, among which is the Post Lyme Disease, whose epidemiological incidence is low, pathophysiology is controversial and the diagnostic approach is based especially on clinical interpretation, directing its therapeutic approach towards symptomatic resolution. We present a case of a female patient of 17 years of age with Lyme disease of recent diagnosis of apparent evolution of 3 years, with previous diagnosis of demyelinating disease, who consulted for neurological deterioration. Scientific considerations on its clinical approach and therapeutic principles are reviewed, contrasting them with those applied.

Passage of Borrelia burgdorferi through diverse Ixodid hard ticks causes distinct diseases: Lyme borreliosis and Baggio-Yoshinari syndrome

Baggio-Yoshinari syndrome is an emerging, tick-borne, infectious disease recently discovered in Brazil. This syndrome is similar to Lyme disease, which is common in the United States of America, Europe and Asia; however, Brazilian borreliosis diverges from the disease observed in the Northern Hemisphere in its epidemiological, microbiological, laboratory and clinical characteristics. Polymerase chain reaction procedures showed that Baggio-Yoshinari syndrome is caused by the Borrelia burgdorferi sensu stricto spirochete. This bacterium has not yet been isolated or cultured in adequate culture media. In Brazil, this zoonosis is transmitted to humans through the bite of Amblyomma and Rhipicephalus genera ticks; these vectors do not belong to the usual Lyme disease transmitters, which are members of the Ixodes ricinus complex. The adaptation of Borrelia burgdorferi to Brazilian vectors and reservoirs probably originated from spirochetes with atypical morphologies (cysts or cell-wall-deficient bacteria) exhibiting genetic adjustments, such as gene suppression. These particularities could explain the protracted survival of these bacteria in hosts, beyond the induction of a weak immune response and the emergence of serious reactive symptoms. The aim of the present report is to note differences between Baggio-Yoshinari syndrome and Lyme disease, to help health professionals recognize this exotic and neglected zoonosis.

Optic neuritis and chorioretinitis as ocular manifestations of borreliosis in Brazil: three cases reported / Neurite óptica e coriorretinite como manifestações oculares da borreliose no Brasil: três casos relatados

Abstract Lyme disease is a systemic infection caused by a tick bite and transmission of the Borrelia burgdorferi spirochete. Species of tick vectors of the disease infest mainly wild or rural animals and rodents that may be asymptomatic reservoirs of the bacteria. Characteristic of the northern hemisphere, Lyme disease in Brazil takes on different characteristics, complicating diagnosis. This paper aims to describe three cases of Lyme-like disease in a city in the state of Bahia, Brazil, with ophthalmologic findings.

Resumo A doença de Lyme é uma infecção sistêmica causada pela picada do carrapato e transmissão da espiroqueta Borrelia burgdorferi. As espécies de carrapatos vetores da doença infestam, principalmente, animais silvestres, rurais e roedores que podem ser reservatórios assintomáticos da bactéria. Característica do hemisfério norte, a doença de Lyme no Brasil assume características distintas, dificultando seu diagnóstico. Esse trabalho tem por objetivo, descrever três casos da doença Lyme símile do Brasil, com achados oftalmológicos, em município do Estado da Bahia.

Brazilian borreliosis with special emphasis on humans and horses

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.

Prevalencia de Borrelia burgdorferi sensu lato en roedores sinantrópicos de dos comunidades rurales de Yucatán, México / Prevalence of Borrelia burgdorferi sensu lato in synanthropic rodents in two rural communities of Yucatán, México

Introducción. La enfermedad de Lyme es una zoonosis multisistémica causada por Borrelia burgdorferi sensu lato. Esta espiroqueta circula en un ciclo enzoótico entre un reservorio vertebrado primario y las garrapatas. Se ha encontrado que varias especies de roedores son eficientes reservorios naturales de B. burgdorferi s.l. Objetivo. Estimar la prevalencia de B. burgdorferi s.l. en roedores sinantrópicos en dos comunidades rurales de Yucatán, México. Materiales y métodos. Se capturaron 123 roedores (94 Mus musculus y 29 Rattus rattus ) para obtener muestras de tejidos de oreja y vejiga. Para la detección de B. burgdorferi s.l. en las muestras, se amplificaron los genes de la flagelina B ( fla B ) y las lipoproteínas de membrana externa, ospC y p66 , mediante reacción en cadena de la polimerasa, y se secuenciaron los amplicones obtenidos. Resultados. La frecuencia de infección por B. burgdorferi s.l. en roedores fue de 36,5 % para flaB (45/123), de 10,5 % (13/123) para p66 y de 3,2 % (4/123) para ospC . En R. rattus la frecuencia de infección fue de 17,2 % y en M. musculus fue de 42,5 %. La frecuencia de infección de B. burgdorferi s.l. en los tejidos estudiados fue de 11,3 % (14/123) en muestras de tejido de vejiga y de 17,0 % (21/123) en las de oreja. No se encontraron diferencias estadísticas (p>0,05) en la frecuencia de infección entre los dos tipos de muestras de tejido utilizadas para el diagnóstico. El gen ospC presentó 98 % de homología con la especie Borrelia garinii , una de las especies heterogéneas del complejo B. burgdorferi s.l. Conclusiones. Los roedores presentaron una alta prevalencia de infección con B. burgdorferi s.l.; las especies M. musculus y R. rattus podrían jugar un papel importante en la continuidad de la presencia de esta bacteria en comunidades rurales de Yucatán, México.

Introduction: Lyme disease is a multisystemic zoonotic disease caused by Borrelia burgdorferi sensu lato. This spirochete circulates in an enzootic cycle between the primary vertebrate reservoir and its tick vectors. Different species of rodents are known to be efficient natural reservoirs for B. burgdorferi s.l. Objective: To estimate the prevalence of B. burgdorferi s.l. in synanthropic rodents from two rural communities of Yucatán, México. Materials and methods: A total of 123 rodents (94 Mus musculus and 29 Rattus rattus ) were trapped, and ear and bladder samples were collected. Flagelin B ( flaB ) genes and outer membrane lipoproteins ospC y p66 were amplified in order to detect B. burgdorferi s.l. presence in the samples. The obtained amplicons were sequenced. Results: The overall infection rates in rodents were 36.5% for flaB (45/123), 10.5% (13/123) for p66, and 3.2% (4/123) for ospC . Rattus rattus had 17.2% of infection and M. musculus , 42.5%. From all examined tissue, 11.3% (14/123) of bladders, and 17.0% (21/123) of ears were infected with the spirochete Borrelia burgdorferi s.l. No statistical differences (p>0.05) were found between the two tissue samples used for diagnosis. The ospC gen was 98% homologous to Borrelia garinii , one species of the B. burgdorferi s.l. complex. Conclusions: We concluded that rodents have a high prevalence of B. burgdorferi s.l. infection, and both species of rodents, M. musculus and R. rattus, might be playing an important role in the maintenance of this bacterium in rural communities of Yucatán, México.

Mielitis transversa aguda en un viajero / Acute transverse myelitis in a traveler

La mielitis transversa aguda se define como un trastorno neuroinmune adquirido de la medula espinal, que ocurre como consecuencia de un evento primario o relacionado a enfermedades inflamatorias autoinmunes, infecciosas o post infecciosas. Entre los agentes etiológicos infecciosos se destaca Borrelia spp., antropozoonosis transmitida por garrapatas de la familia ixodidae. Los pacientes con enfermedad de Lyme desarrollan, entre un 10 a un 15%, manifestaciones neurológicas. El espectro clínico suele ser variado e incierto. Entre las manifestaciones clínicas de la neuroborreliosis de Lyme, la mielitis transversa aguda ha sido reportada entre el 4 al 5%. Se describe el caso de un viajero proveniente de zona endémica de enfermedad de Lyme con encefalomielitis secundaria a infección aguda por Borrelia burgdorferi que presentó resolución completa de los síntomas luego de finalizar el tratamiento antibiótico.

Acute transverse myelitis is defined as an acquired neuroimmune disorder of the spinal cord, which occurs as a consequence of a primary event, or directly related to an autoimmune inflammatory disease, an infectious or post-infectious disease. Amongst infectious etiologies, Borrelia spp., a tick-bourne anthropozoonosis of the ixodidae family, prevails. Approximately 10 to 15% of patients with Lyme disease undergo neurologic manifestations, with an assorted and uncertain array of clinical syndromes. Transverse myelitis accounts for up to 5% of Lyme neuroborreliosis. We describe the case of a traveler from endemic zone for Lyme disease, with encephalomyelitis secondary to acute infection by Borrelia burgderfori, with complete resolution of symptoms after concluding adequate antibiotic treatment.

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.

Estudo biomolecular de produtos de Chlamydophila pneumoniae, Mycoplasma pneumoniae e Borrelia burgdorferi na etiopatogenia da degeneração mixomatosa da valva mitral / A biomolecular study on Chlamydophila pneumoniae, Mycoplasma pneumoniae and Borrelia burgdorferi products in myxoid mitral valve degeneration etiopathogenesis

Introdução e Objetivo: A doença mixomatosa da valva mitral leva ao comprometimento de sua matriz devido à alteração em sua composição tecidual provocada pelo desequilíbrio na quantidade de ácidos mucopolissacarídeos ou glicosaminoglicanos. Sua etiologia ainda não está totalmente esclarecida, podendo ocorrer em formas familiares com transmissão autossômica dominante de penetrância variável, que pode ser dependente do tempo ou de prováveis fatores ambientais, situações em que a interação de agentes infecciosos necessita de maiores esclarecimentos. O objetivo deste estudo é a análise dos produtos dos patógenos da Chlamydophila pneumoniae, Mycoplasma pneumoniae e Borrelia burgdorferi em segmentos de cúspide retirados da valva mitral com degeneração mixomatosa, comparada ao grupo controle e a relação dos produtos bacterianos com aumento de marcadores inflamatórios (CD20, CD48, CD68) e de metaloproteinase (MMP9) na etiopatogenia da degeneração mixomatosa da valva mitral. Método: Estudo observacional, analítico, tipo caso-controle, que analisou 2 grupos contendo 20 pacientes cada e divididos em grupo 1, composto por fragmentos de tecido valvar mitral com diagnóstico de degeneração mixomatosa extraídos em procedimentos de troca ou plásticas valvares mitrais; e grupo 2, formado por segmentos de valvas mitrais sem valvopatia retirados no serviço de verificação de óbito. Foram realizadas colorações de hematoxilina e eosina e Movat para diagnóstico histológico da degeneração mixomatosa e técnica de imunohistoquímica para detecção de antígenos da Borrelia burgdorferi, Mycoplasma pneumoniae, mediadores inflamatórios (CD20, CD45, CD68) e marcadores de metaloproteinase (MMP9). A presença de antígenos da Chlamydophila pneumonia e foi pesquisada pela técnica de hibridização in situ. A análise quantitativa dos aspectos microscópicos foi realizada com o analisador de imagens Aperio. A pesquisa de elementos bacterianos foi feita através de microscopia eletrônica...

Background: The myxomatous mitral valve disease leads to impairment due to changes in their tissue composition caused by the imbalance in the amount of acid mucopolysaccharides or glycosaminoglycans. Its etiology is not yet fully understood and may occur in familial forms of autosomal dominant trait with variable penetrance that can be time-dependent or probable environmental factors, where the interaction of infectious agents requires further elucidation. The purpose of this study is the analysis of the pathogens products of Chlamydophila pneumoniae, Mycoplasma pneumoniae and Borrelia burgdorferi in removed cusp segments of the mitral valve with myxoid degeneration, compared to the control group and the ratio of bacterial products with increased inflammatory markers (CD20, CD48, CD68) and metalloproteinase (MMP9) in the pathogenesis of myxomatous degeneration of the mitral valve. Method: Observational, analytical, case-control study which analyzed 2 groups of 20 patients each and divided in group 1, consisting of fragments of mitral valve tissue with diagnosis of myxomatous degeneration extracted in replacement procedures or mitral valve repair; group 2, formed by segments of mitral valves without valvolpaty clinial disease removed in the coroner service. Hematoxylin and eosin and Movat stains were done for histological diagnosis of myxoid degeneration and immunohistochemical technique for the detection of Borrelia burgdorferi, Mycoplasma pneumonia antigens, inflammatory mediators (CD20, CD45, CD68) and markers of metalloproteinase (MMP9). The presence of Chlamydophila pneumonia antigens was verified through an in situ hybridization technique. The quantitative analysis of the microscopic aspects was performed with the Aperio image analyzer. The research of bacterial elements was performed by a transmission electron microscopy. Results: In group 1, 14 (70%) patients were male and 6 (30%) were female. The mean age was (51 to 79 years, sd...

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.

Spatial distribution of vector borne disease agents in dogs in Aegean region, Turkey / Distribución espacial de agentes patógenos transmitidos por vectores en perros en la región del Egeo, Turquía

Objective. Assess the spatial distribution of seroprevalence of infection with or exposure to 4 vector-borne pathogens Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi and Dirofilaria immitis, across the coastal states of the Aegean region with special reference to clinical signs and haematological variances related to disease condition. Materials and methods. A convenience sample, targeting blood from at least 10 pet dogs from Izmir, Aydin, Denizli, Mugla and Manisa cities involved was evaluated using a canine point-of-care ELISA kit. Results. Out of 307 dogs tested the overall seroprevalence was highest for E. canis (24.42%), followed by E. canis + A. phagocytophilum co-infection (10.42%), A. phagocytophilum (7.49%) and D. immitis (2.28%). Only 2 cases were seropositive to B. burgdorferi albeit 10 dogs were co-infected with more than 2 agents. For both dogs infected with E. canis and co-infected with E. canis and A. phagocytophilum, anemia, thrombocytopenia and leukocytosis, were more commonly detected, whereas thrombocytopenia and leukocytosis were significant finding in dogs infected with A. phagocytophilum or D. immitis, respectively. Variance analysis showed significant differences for mean RBC, Hb, PCV and PLT values (p<0.01) among control group and other groups. Conclusions. Seropositivity for vector-borne pathogens other than B. burgdorferi, is moderately to widely distributed in dogs residing in the Aegean region in Turkey.

Objetivo. Evaluar la distribución espacial de la seroprevalencia de la infección de 4 agentes patógenos de transmisión por vectores Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi y Dirofilaria immitis, en los estados costeros de la región del Egeo con especial referencia a los signos clínicos y las variaciones hematológicas relacionadas con la enfermedad. Materiales y métodos. Se tomaron por conveniencia muestras de sangre de al menos 10 perros en las ciudades Izmir, Aydin, Denizli, Mugla y Manisa. Para la evaluación de las muestras se utilizó un kit de ELISA para la detencción de anticuerpos de las enfermedades del estudio. Resultados. De los 307 perros muestreados, la seroprevalencia más alta fue para E. canis (24.42%), seguido por la coinfección entre E. canis + A. phagocytophilum (10.42%), A. phagocytophilum (7.49%) y D. immitis (2.28%). Sólo 2 casos fueron seropositivos para B. burgdorferi aunque 10 perros fueron coinfectados con más de 2 agentes. En ambos perros infectados con E. canis y coinfectados con E. canis y A. phagocytophilum, se detectó comúnmente anemia, trombocitopenia y leucocitosis, mientras que la trombocitopenia y leucocitosis fueron significativos en perros infectados con A. phagocytophilum o D. immitis , respectivamente. El análisis de varianza mostró diferencias significativas para los promedios de RBC, hemoglobina, hematocrito y valores PLT (p<0.01) entre el grupo control y los otros grupos. Conclusiones. La seropositividad transmitida por vectores patógenos distintos de B. burgdorferi, fue moderada y ampliamente distribuida en los perros que residen en la región del Egeo en Turquía.

Atrofoderma de Pasini y PieriniPosible variante de la esclerodermia localizada / Pasini and Pierini atrofoderma Possible variant of localized scleroderma

Resumen Reportamos el caso de una paciente con unas lesiones en piel que fueron diagnosticadasinicialmente como morfea, por lo cual fue remitida al servicio de reumatología. Las características clínicas e histológicas nos permitieron descartar morfea clásica y clasificarla mejor como una atrofoderma de Pasini y Pierini. Aunque algunos la consideran una variante localizada y benigna dela morfea para otros se trata de una entidad nosológica distinta. Esta enfermedad debe ser considerada en el diagnóstico diferencial de la esclerodermia localizada y todos aquellos desórdenes que semejan esclerodermia para lograr un diagnóstico correcto y evitar tratamientos y exámenes innecesarios.Informamos la ausencia de respuesta de sus lesiones con el uso de hidroxicloroquina. (Acta MedColomb 2014; 39: 77-80).

Abstract The case of a patient with skin lesions initially diagnosed as morphea is reported. With this diagnosis, she was referred to rheumatology. The clinical and histological characteristics allowed us to rule out classical morphea and better classify it as a Pasini and Pierini atrofoderma. While some consider it a localized and benign variant of morphea, for others it is a distinct nosological entity. This disease should be considered in the differential diagnosis of localized scleroderma and all those disorders that resemble scleroderma for a proper diagnosis and to avoid unnecessary tests and treatments. The lack of response from her lesions with the use of hydroxychloroquine is reported. (Acta Med Colomb 2014; 39: 77-80).

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.

Ensayo de infección artificial de garrapatas cubanas con Borrelia burgdorferi sensu stricto / Artificial infection assay of Cuban ticks with Borrelia burgdorferi sensu stricto

Como es conocido, Borrelia spp. constituye un grupo importante de patógenos bacterianos transmitidos por garrapatas, a excepción de Borrelia recurrentis que es transmitida por piojos del cuerpo. Estos microorganismos son los causantes de borreliosis o enfermedad de Lyme y fiebre recurrente.1El complejo B. burgdorferi sensu lato es causante de la enfermedad de Lyme, y el principal vector descrito lo constituyen las garrapatas duras del género Ixodes.2 Recientemente se ha reportado en México y Brasil la presencia de ADN de B. burgdorferi sensu stricto,3,4 una de las 19 genomaespecies del complejo, en garrapatas Amblyomma cajennense, pero se desconoce si esta especie es capaz de transmitir las borrelias a un nuevo hospedero. En Cuba, a partir de evidencias serológicas específicas de la infección por este agente y teniendo en cuenta las especies de ixódidos que prevalecen, así como sus características ecológicas y antropofílicas, las garrapatas A. cajennense y Rhipicephalus sanguineus se consideran vectores potenciales de las borrelias.5Sobre la base de los hallazgos e hipótesis anteriormente mencionados se hace necesario iniciar investigaciones para comprobar si otras especies diferentes de Ixodes spp. son capaces de transmitir las borrelias. Ello requiere contar con un método sencillo y eficaz que permita la infección artificial de ixódidos. Se han reportado disímiles estrategias con estos fines, entre las que se incluyen el empleo de cámaras de alimentación con membranas naturales o artificiales, la infusión por enema, la inoculación hemocélica y la alimentación por capilares...

Primary embryonic cells of Rhipicephalus microplus and Amblyomma cajennense ticks as a substrate for the development of Borrelia burgdorferi (strain G39/40) / Células primárias embrionárias do carrapato Rhipicephalus microplus e Amblyomma cajennense como substrato para desenvolvimento de Borrelia burgdorferi (Cepa G39/40)

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.

Borrelia burgodorferi, o agente da borreliose de Lyme, é uma espiroqueta transmitida por carrapatos aos seres humanos e animais. Seu cultivo in vitro em células de carrapato permite estudos de sua biologia e propicia metodologia para futuras pesquisas no Brasil, para o isolamento de Borrelia spp. Nós examinamos in vitro as características de células embrionárias de Rhipicephalus microplus e Amblyomma cajennense, e a viabilidade de utilização dessas células embrionárias como um substrato para cultivo de B.burgdorferi. Subculturas foram preparadas a partir de culturas primárias de células embrionárias de R. microplus e A. cajennense mantidas em meio Leibovitz's (L-15) completo, a 28 ºC e 31 ºC, respectivamente. Com a formação da monocamada, o L-15 foi substituído pelo meio Barbour-Stoener-Kelly, para o experimento de infecção com B. burgdorferi nas culturas de células. Após 72 horas de cultivo, realizou-se a contagem das espiroquetas, as quais foram avaliadas sob microscópio invertido de contraste de fase e campo escuro (400×). Verificou-se a sobrevivência, a multiplicação e a aderência de B. burgdorferi em células embrionárias de R. microplus e A. cajennense. No estudo da cultura de B. burgdorferi com células embrionárias de R. microplus, observou-se, na contagem final, média de 2,4 × 10(7) espiroquetas/mL; no cultivo livre de células, verificou-se média de 2,5 × 10(7) espiroquetas/mL. No cultivo de A. cajennense, a contagem final de espiroquetas foi, em média, 1,7 × 10(7) espiroquetas/mL, enquanto que, para as cultivadas livres de células, se verificou média de 2,2 × 10(7) espiroquetas/mL. Resultado semelhante foi observado na contagem final de espiroquetas cultivadas em células de R. microplus e A. cajennense, quando comparado com o controle livre de células. Estes resultados demonstraram que células de R. microplus e A. cajennense têm o potencial para serem utilizadas como substrato para o crescimento de B. burgdorferi no estudo da interação com as células do hospedeiro.

Prevalence of antibodies to Borrelia burgdorferi sensu stricto in humans from a Cuban village

Lyme disease has not been officially reported in Cuba. However, clinical cases have been serologically reported. Seroprevalence survey of Borrelia burgdorferi sensu stricto antibodies in humans in the country has not been conducted. OBJECTIVE: To estimate the prevalence of borrelial antibodies in inhabitants of a village with historically high level of tick infestation. METHODS: Serum specimens from 247 persons randomly selected from the population of the village were examined by IgG Western blot using B31 strain for estimating the prevalence of antibodies profile. RESULTS: A seroprevalence value interval (95 percent CI) of 0.6 percent-7.2 percent was estimated for the studied population. The prevalent borrelial protein bands on immunoblots were 41, 72, 90/93, 34, 47, 60, 58, 56, 65/66 and 31 kDa in a decreasing order of significance. CONCLUSION: These results support the previous serological findings, suggesting the presence of this borreliosis in Cuba.

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

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 (Boophilus)microplus, 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 contra B. burgdorferi, com positividade variando de 63,6% a 92,9%. O alto número de soropositivos pode ser explicado pela frequente presença do carrapato Rhipicephalus (Boophilus) microplus e pela possível existência de espiroquetas do gênero Borrelia infectando búfalo na região estudada, embora novos estudos sejam necessários para a confirmação desta relação. Estes fatos sugerem reação cruzada entre a cepa americana G39/40 de B. burgdorferi utilizada como substrato antigênico e a espécie de Borrelia spp. que possivelmente infecta bubalinos no estado do Pará.