Development of IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro / Desenvolvimento de Anticorpos IgY em galinhas e IgG em coelhos imunizados contra proteínas de Pythium insidiosum isolado de equinos no Estado do Rio de Janeiro, Brasil

Pythiosis is caused by Pythium insidiosum and the occurrence of disease in horses was described in the North and Northwest State of Rio de Janeiro, Brazil. The disease was described in cattle, sheep, humans, and horses in different states and regions across the country. This paper describes the development of IgY and IgG polyclonal antibodies, in chicken and rabbits, respectively against proteins extracted from kunkers and hyphae of P. insidiosum from affected horses. The proteins were recognized by chicken, rabbit and horse antibodies by immunodiffusion and Western blot against majority bands of 27 and 43 KDa, and titrated by ELISA. The antibodies IgY developed by the first time against Brazilian strains of P. insidiosum may represent a valuable tool in the detection of antigens of the pathogen and contribute to further studies aimed at immunotherapy and knowledge about this disease in endemic areas in Rio de Janeiro and in Brazil.

Pitiose é causada por Pythium insidiosum e a doença foi descrita em equinos no Norte e Noroeste do Estado do Rio de Janeiro, Brasil. A doença foi descrita em bovinos, ovelhas, humanos e cavalos em diferentes estados e regiões do país. Este trabalho descreve o desenvolvimento de anticorpos policlonais, IgY e IgG, em galinha e coelho, respectivamente, contra proteínas extraídas de kunkers e hifas de P. insidiosum de cavalos doentes. As proteínas foram reconhecidas por anticorpos de galinha, coelho e cavalos contra as bandas majoritárias de 27 e 34 KDa em imunodifusão e Western blot tituladas por ELISA. Os anticorpos IgY desenvolvidos pela primeira vez contra cepas brasileiras de P. insidiosum podem representar um valioso instrumento na detecção de antígenos de patógenos e contribuem para novos estudos baseados na imunoterapia e no entendimento sobre esta doença em áreas endêmicas no Rio de Janeiro e em todo o país.

Detection of different Staphylococcus aureus strains in bovine milk from subclinical mastitis using PCR and routine techniques

Contamination of fresh milk with Staphylococcus aureus was assessed comparatively through routine phenotypic (coagulase tube test and coagulase slide test) and genotypic (PCR) screening of 128 S. aureus strains isolated from 555 milk samples. These samples were collected from 362 cows with subclinical mastitis, hosted in different dairy herds at various locations of the Northern and Northeastern rural areas of the State of Rio de Janeiro, 39.7 per cente of which were CMT-positive. All S. aureus isolates tested positive for the presence of the coagulase gene by PCR and the isolates could be grouped into four distinct classes according to the size of the PCR product. The strains also yielded variable results when assayed with coagulase test. Taken together, these data indicate the existence of extensive polymorphism at the coagulase gene locus in the genus Staphylococcus and exemplifies the extent of molecular and phenotypic heterogeneity associated with the strains circulating in rural herds.(au)