ABSTRACT
Abstract INTRODUCTION: We investigated the occurrence of relapsing fever (RF) causing Borrelia genus spirochetes in synanthropic bats from the municipality of Maringá, Paraná, South of Brazil. METHODS: Tissue samples from the wings of bats were collected monthly from April 2013 to February 2014 and extracted DNA was used to evaluate the presence of RF causing Borrelia spp. RESULTS: All bat tissues tested negative for RF causing Borrelia spp. CONCLUSIONS: Borrelia spp. do not occur in chiropterans from Maringá.
Subject(s)
Animals , Borrelia/isolation & purification , Chiroptera/microbiology , Time Factors , Brazil , DNA, Bacterial , Forests , Polymerase Chain Reaction , Sequence Analysis, DNA , Electrophoresis, Agar GelABSTRACT
Abstract This study aimed to perform a morphological, molecular and phylogenetic characterization of Borrelia theileri obtained from infected Rhipicephalus microplus in Brazil. Fifty engorged R. microplus females from cattle in the municipality of Seropédica, Rio de Janeiro, were analyzed for spirochetes by hemolymph smear. Macerated eggs and positive ticks, as well as blood from the bovine infested by these ticks, were analyzed the glpQ, flaB and hpt genes by PCR. The PCR products were purified and sequenced for analysis and construction of a phylogenetic tree. Only 2% (1/50) of the ticks generated a positive result by both smear and PCR. The spiral forms (n = 50) had (media ± SD) a mean length of 19.17 ± 4.12 µm, diameter of 0.2935 ± 0.0469 and number of turns 8.44 ± 2.59. Sequence alignments of the three evaluated genes exhibited 98% similarity to B. theileri isolates, occurring in a clade highly related to B. theileri strain KAT. Egg maceration samples were positive for the three evaluated genes, whereas bovine blood was negative by PCR. This is the most detailed characterization of B. theileri in the Americas to-date, presenting morphological, molecular and phylogenetic data, including the transovarial transmission of the spirochete in the host tick.
Resumo O estudo teve como objetivo realizar a caracterização morfológica, molecular e filogenética de Borrelia theileri obtida de Rhipicephalus microplus naturalmente infectado em bovino no estado do Rio de Janeiro, Brasil. Um total de 50 fêmeas de R. microplus ingurgitadas foram analisadas para espiroquetas por meio de esfregaço de hemolinfa. Ovos macerados e carrapatos, assim como sangue de bovinos infectados por esses carrapatos, foram analisados os genes glpQ, flaB e hpt por PCR. Os produtos de PCR foram purificados e sequenciados para análise e construção de uma árvore filogenética. Apenas 2% (1/50) dos carrapatos geraram um resultado positivo tanto pelo esfregaço como pela PCR. As formas espirais (n = 50) apresentaram (média ± DP) comprimento médio de 19,17 ± 4,12, diâmetro de 0,2935 ± 0,0469 e número de voltas de 8,44 ± 2,59. Os alinhamentos das sequências dos três genes avaliados exibiram 98% de similaridade aos isolados de B. theileri, ocorrendo em um clado altamente relacionado à linhagem de B. theileri KAT. As amostras de maceração de ovos foram positivas para os três genes avaliados, enquanto o sangue bovino foi negativo pela PCR. Esta é a mais completa caracterização de B. theileri nas Américas, apresentando dados morfológicos, moleculares e filogenéticos, incluindo a transmissão transovarial da espiroqueta no carrapato hospedeiro.
Subject(s)
Animals , Female , Borrelia/genetics , Rhipicephalus/microbiology , Phylogeny , Borrelia/isolation & purification , Cattle/parasitology , Polymerase Chain ReactionABSTRACT
Relapsing fever is an acute febrile illness caused by spirochetes of the genus Borrelia. The high fevers of presenting patients spontaneously abate and then recur. Here we report a 50-year-old woman having relapsing fever associated with thrombocytopenia. Giemsa staining of peripheral blood smear revealed spiral organisms morphologically resembling Borrelia. A rare case of relapsing fever which was successfully treated with doxycycline is discussed.
Subject(s)
Animals , Anti-Bacterial Agents/therapeutic use , Arachnid Vectors/microbiology , Borrelia/isolation & purification , Doxycycline/therapeutic use , Female , Humans , India , Insect Vectors/microbiology , Phthiraptera/microbiology , Middle Aged , Ornithodoros/microbiology , Relapsing Fever/diagnosisABSTRACT
O presente trabalho teve como objetivo reportar a ocorrência de Borrelia spp. em culturas de células embrionárias de Boophilus microplus infectados naturalmente. Sete dias após o início de uma nova cultura primária de células embrionárias do carrapato B. microplus, incubadas a 31ºC, notou-se que as células começaram a degenerar. Ao exame em microscópio de contraste de fase detectou-se a presença de microrganismos alongado e com grande mobilidade. Lâminas de microscópio confeccionadas com amostras do sobrenadante da cultura, hemolinfa e massa de ovos, coradas pelo May Grünwald-Giemsa, permitiram a visualização de espiroquetas. O exame morfológico do microrganismo e sua visualização em B. microplus sugere ser Borrelia spp.
The aim of the present work was to report the occurrence of Borrelia spp. in embryonic cell cultures from naturally infected Boophilus microplus. Seven days after the beginning of a primary culture of embryonic cells of B. microplus at 31ºC was noted that the cells start suffering degeneration. Under examination at phase contrast microscope, the presence of prolongated microorganisms with great mobility was detected. Microscopic slides of the culture supernatant, hemolymph and egg mass, were stained by May Grünwald-Giemsa, allowing the visualization of the spirochetes. The morphologic examination of the microorganism and its visualization in. B. microplus, suggest to be Borrelia spp.
Subject(s)
Animals , Borrelia/isolation & purification , Ixodidae/cytology , Ixodidae/embryology , Cells, Cultured/microbiologyABSTRACT
Forty-four marsupials, 77 rodents and 161 ticks were captured in an Atlantic Forest Reserve in Cotia county, State of Sao Paulo, where human cases of Lyme disease (LD) simile were reported. Twenty-one borrelia-like spirochete isolates were recovered from the mammals' blood and rodent livers or spleens, and triturated ticks inoculated into BSK II medium. Our results suggest that the reservoirs and ticks collected may harbor borrelia-like spirochetes, some of which have an antigenic similarity with the unknown causative agent of LD simile in Brazil, and/or with North American Borrelia burgdorferi s.s.
Subject(s)
Humans , Animals , Borrelia/isolation & purification , Marsupialia/microbiology , Rodentia/microbiology , Ticks/microbiology , Trees , Arachnid Vectors/microbiology , Brazil , Disease Reservoirs , Lyme Disease/transmissionABSTRACT
Lyme disease is caused by the spirochete, Borrelia burgdorferi, a bacteria which infects many vertebrates including humans. Borrelia have been isolated from many parts of the world, and there is interest to identify commun genetic markers to improve molecular methods of diagnosis, and to aid in understanding varied manifestations of the disease. A total of 48 Borrelia burgdorferi strains, including: 38 isolated from ticks (Ixodes dammini, I. persulcatus, I. ricinus and I. pacificus), 3 from animals (dog, bird and hamster), and 7 from human clinical cases (skin, CSF, plasma and blood) from different geographic areas, were studied by DNA/DNA hybridization and rRNA gene restriction patterns by using a biotinylated pKK3535 probe (Altewegg M., Mayer L.W., 1989). The migration patterns of rRNA generestriction fragments after clevage by Hind III separate these strains into 5 ribotypes of Borrelia burgdorferi: Type I (38 American, 2 European strains); Type II (13 American strains); Type III (3 Asian and 1 European strains); Type IV (1 European and 2 Asian strains) and Type V (1 Asian strain). The use of ribotyping has provided and additional tool to investigate the diferences or commun patterns which cause various Lyme disease syndromes.
Subject(s)
Borrelia/classification , Borrelia/genetics , Borrelia/isolation & purification , Lyme Disease/diagnosis , Lyme Disease/epidemiology , Lyme Disease/etiology , RNA, Ribosomal/genetics , Syndrome , Molecular EpidemiologyABSTRACT
Se describe un nuevo medio solido para la rapida deteccion de Borrelia persica y Borrelia microti. Corrientemente el cultivo y aislamiento de Borrelia demora alrededor de 21 dias. El examen serologico mas frecuentemente realizado demora menos tiempo pero esta asociado con resultados falsos positivos relativamente altos. Sin embargo, nuestro nuevo medio solido reduce el tiempo de cultivo a 72 horas, lo que permite un rapido diagnostico de la enfermedad causada por Borrelia persica y Borrelia microti y el inicio temprano del tratamiento en estos pacientes