Validação de nova proposta de espermocultura quantitativa aplicada a sêmen industrializado de touros / Validation of a new proposal of quantitative sperm culture for industrialized bull semen

O Ministério da Agricultura, Pecuária e Abastecimento (MAPA) tem discutido a obrigatoriedade da realização de espermocultura em sêmen industrializado não somente para garantia da biossegurança mas também por falhas na fertilização in vitro pela contaminação dos ovócitos com bactérias ubiquitárias e oportunistas da microbiota prepucial. A técnica quantitativa de Pour Plate, preconizada pela Organização Internacional de Epizootias, é operacionalmente difícil e de custo elevado para a rotina em centrais de inseminação artificial (CIAs). Desse modo, avaliou-se a técnica de contagem de bactérias viáveis por superfície (CBVS) em UFC/mL de sêmen industrializado de touros de CIAs, comparando-a à técnica de Pour Plate, com intuito de validação. Foram empregadas palhetas de sêmen de touros do Projeto Hungria do MAPA. A partir da diluição seriada de 10-1 a 10-5 do sêmem, foram empregadas, paralelamente, as técnicas de Pour Plate e CBVS. A média ou mediana de UFC/mL obtida em cada diluição foi comparada entre as técnicas, e não foram observadas diferenças estatísticas entre as duas técnicas quanto à quantificação de bactérias em UFC/mL, sugerindo a possibilidade de substituição da técnica de Pour Plate pela de CBVS, com vantagens de praticidade e menor custo.

The Brazilian Ministry of Agriculture (MAPA) has discussed the mandatory culture of industrialized semen, both to ensure biosefety, and to prevent in vitro fertilization problems caused by oocyte contamination with ubiquitous and opportunistic bacteria from preputial microbiota. Pour plate, a quantitative technique recommended by the World Organization for Animal Health (OIE), is operationally difficult and costly for routine analysis in Artificial Insemination Centers (AICs). The objectives of this study were to evaluate and validate viable superficial bacteria counts (VSBC), in CFU/mL, compared with pour plate counts, in industrialized bull semen samples from AICs. Semen straws from Projeto Hungria - MAPA bulls were used. VSBC and pour plate were carried out in parallel in serial dilutions of the samples, from 10-1 to 10-5. CFU/mL means or medians recorded in each dilution and technique were compared, and no statistical differences were observed between the two techniques regarding the quantification of bacteria in CFU/mL, suggesting that it may be possible to replace pour plate for CBSV, a cheaper and more practical technique.

Enumeration of heterotrophic bacteria in water for dialysis: comparison of the efficiency of Reasoner'2 agar and plate count agar

Due to the fact the incubation conditions may influence the microbiological evaluation of water for dialysis, the objective of the present study was the comparison of the efficiency of R2A and PCA media in the enumeration of heterotrophic bacteria in 193 samples of water collected in dialysis clinics from 12 cities in São Paulo, between October and December 2007. Results showed counts significantly greater in R2A, suggesting that enumeration should be carried out in R2A, suggesting that enumeration should be carried out in R2A agar associated with longer incubation times, because of the greater sensitivity.