Characterisation of the methylation pattern in the intragenic CpG island of the IGF2 gene in Bos taurus indicus cumulus cells during in vitro maturation

The aim of this study was to characterise the methylation pattern in a CpG island of the IGF2 gene in cumulus cells from 1-3 mm and a parts per thousand yenaEuro parts per thousand 8.0 mm follicles and to evaluate the effects of in vitro maturation on this pattern.Genomic DNA was treatment with sodium bisulphite. Nested PCR using bisulphite-treated DNA was performed, and DNA methylation patterns have been characterised.There were no differences in the methylation pattern among groups (P > 0.05). Cells of pre-IVM and post-IVM from small follicles showed methylation levels of 78.17 +/- 14.11 % and 82.93 +/- 5.86 %, respectively, and those from large follicles showed methylation levels of 81.81 +/- 10.40 % and 79.64 +/- 13.04 %, respectively. Evaluating only the effect of in vitro maturation, cells of pre-IVM and post-IVM COCs showed methylation levels of 80.17 +/- 12.01 % and 81.19 +/- 10.15 %.In conclusion, the methylation levels of the cumulus cells of all groups were higher than that expected from the imprinted pattern of somatic cells. As the cumulus cells from the pre-IVM follicles were not subjected to any in vitro manipulation, the hypermethylated pattern that was observed may be the actual physiological methylation pattern for this particular locus in these cells. Due the importance of DNA methylation in oogenesis, and to be a non-invasive method for determining oocyte quality, the identification of new epigenetic markers in cumulus cells has great potential to be used to support reproductive biotechniques in humans and other mammals.

Ocorrência da infecção por Cryptosporidium spp. em cabritos (Capra hircus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detecção do HPV por nPCR em leucoplasias bucais: estudo caso-controle

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização molecular de Cryptosporidium spp. em bezerros (Bos taurus e Bos indicus) no município de Formiga, Minas Gerais - Brasil

A criptosporidiose, é a enfermidade de veiculação hídrica, possui como agravante a dificuldade de prevenção da contaminação ambiental e ausência de medidas terapêuticas eficazes. Com acentuada importância na bovinocultura, ocasiona inflamação e atrofia das vilosidades intestinais resultando em perda da superfície de absorção. Este estudo teve como objetivo realizar a caracterização molecular da infecção por Cryptosporidium spp. em bezerros do Município de Formiga, Minas Gerais. Um total de 300 amostras de fezes de bezerros holandeses, Nelore e sem raça definida saudáveis foram avaliadas pela técnica de coloração contraste negativo de verde malaquita e por meio da reação de Nested-PCR para amplificação de fragmentos de DNA da subunidade 18S do gene do RNA ribossômico. Ocorrência de 5,33% (16/300) pelo verde malaquita e 4,66% (14/300), pela PCR foi observada, sendo que nenhuma correlação foi verificada entre a positividade e as variáveis estudadas. Por meio da caracterização molecular foram identificadas as espécies Cryptosporidium andersoni e Cryptosporidium ryanae. Como conclusão, observou-se baixa ocorrência da infecção e eliminação de oocistos por Cryptosporidium spp, ausência de sinais clínicos nos animais, houve forte concordância entre os resultados obtidos por meio das duas técnicas utilizadas e pela caracterização molecular (Nested-PCR) foram diagnosticadas as espécies C. andersoni e C. ryanae, presentes em faixas etárias não relatadas na literatura. Estas duas espécies de Cryptosporidium supracitadas são descritas pela primeira vez, parasitando bovinos no estado de Minas Gerais.

Diferentes métodos de diagnóstico molecular na diferenciação das espécies de Sarcocystis spp

The Sarcocystis genus includes obligatory two-host life cycle protozoan parasites. It is the most numerous of the six genera of the Sarcocystidae family. The infection caused by parasites of this genus is a zoonotic and cosmopolitan disease known as sarcosistosis or sarcosporidiosis. The sarcositosis though frequently asymptomatic in its definitive hosts can be fatal in its intermediate hosts. The usual diagnoses of sarcosistosis takes place through a histological demonstration of schizonts in blood vessels and organs, and the presence of cysts in muscle tissue by necropsy or biopsy, this second method still more common and based on morphological features of the sarcocyst. However, these methods can be inadequate to a precise identification of the infector species once that, besides the genus being of numerous species, these often present similar morphological features. Another factor that makes the diagnostic more difficult is the non specificity of some Sacocystis species to their hosts. Consequently, molecular diagnostic methods have been used in order to identify the infector species and the parasite specific biological cycles, demonstrating also new species and coevolutive aspects between parasite and host. Among the most employed molecular techniques the Polimerase Chain Reaction (PCR), the nested-PCR and the Restriction Fragment Length Polymorphism (RFLP) stands out

Adesão de Paracoccidioides brasiliensis e outros fungos anemófilos a teias de aranhas: implicações para a identificação de nichos microbianos no ambiente

The dimorphic pathogenic fungus, Paracoccidioides brasiliensis (Pb), is the etiological agent of paracoccidioidomycosis (PCM), the most important systemic mycosis in Latin America. While the yeast phase can be isolated from patients affected by paracoccidioidomycosis, dogs and naturally infected armadillos; several elements related to the ecology of the saprophytic phase of the pathogen, which is responsible for the production of infective propagules, are poorly understood, hampering the adoption of preventive measures. The demonstration of the high incidence of Pb infection in the 9- banded armadillo, Dasypus novemcinctus, has opened new perspectives for the identification of the pathogen’s habitat. At the opening of the armadillos’ burrows, spider webs are commonly found. The objective of this study was to evaluate the presence of Pb in spider webs samples related to the habitat of armadillos. Spider web samples were collected at Lageado Farm, Botucatu/SP and prepared for microscopic, molecular and mycological analyses. Microscopic analysis showed that different fungi were closely attached to spider web samples. Nested-PCR reaction showed positive amplification for Pb in 4 samples, with identity confirmed by amplicon sequencing. Fungal colonies also included members of Aspergillus, Blastobotrys, Penicillium, Candida, and Sporothrix genera, which are related to opportunistic disease and primary infections of great medical importance. In vitro adhesion tests of mycelia and yeast form of Pb to the spider webs were also performed, in order to analyze the possible physical attraction between fungal cells and the spider web protein network. The results showed a clear adherence of fungal particles to spider webs. In the current literature, there are no studies reporting adhesive properties of microorganisms to spider webs... (Complete abstract click electronic access below)

Ocorrência e caracterização molecular de Cryptosporidium spp. em caprinos jovens do município de Quixadá-Ceará

Pós-graduação em Medicina Veterinária - FCAV

Infecção por Cryptosporidium em bezerros e humanos de assentamentos rurais na região de Araçatuba, São Paulo

Pós-graduação em Medicina Veterinária - FCAV

Pesquisa de Toxoplasma gondii (Nicolle & Manceaux, 1909) em sêmen, testículos e epidídimos de gatos (Felis catus) primoinfectados

Pós-graduação em Medicina Veterinária - FCAV

Molecular characterisation of Cryptosporidium spp. in lambs in the south central region of the State of São Paulo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Vigilância do vpirus da dengue e circulação dos sorotipos presentes em São José do Rio Preto entre 2011 e 2014

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparação entre métodos diagnósticos da tuberculose em bovinos abatidos em matadouros-frigoríficos do Estado de São Paulo

Pós-graduação em Medicina Veterinária - FCAV

Serum DHEA-S increases in dogs naturally infected with Ehrlichia canis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Humoral immune responses against the malaria vaccine candidate antigen Plasmodium vivax AMA-1 and IL-4 gene polymorphisms in individuals living in an endemic area of the Brazilian Amazon

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular and serological detection of Babesia Bovis and Babesia Bigemina in cattle in the Rio de Janeiro, Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)