Navegando por Autor "Ribon, A. O. B."
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Item Cloning and characterization of a gene encoding the endopolygalacturonase of Penicillium griseoroseum(Biotechnology Letters, 1999-05) Ribon, A. O. B.; Coelho, J. L. C.; Barros, E. G. de; Araújo, E. F.A conserved region of a polygalacturonase (PG) gene from Penicillium griseoroseum was PCR amplified and used to screen a genomic library from this fungus. The nucleotide sequence of the isolated clone (pggI) consisted of 1497 bp, including a coding region of 1251 bp. This region potentially encodes a protein of 376 amino acids, and is interrupted by two introns. Extensive homology was observed between this protein and several fungal endopolygalacturonases. DNA hybridization analyses revealed that there is a low copy number of pggI in the P. griseoroseum genome, probably one or two copies.Item Differential expression of polygalacturonase-encoding genes from Penicillium griseoroseum in different carbon sources(Journal of Industrial Microbiology and Biotechnology, 2002-06-14) Ribon, A. O. B.; Queiroz, M. V.; Coelho, J. L. C.; Araújo, E. F deA second polygalacturonase - encoding gene ( pgg2 ) of Penicillium griseoroseum was cloned and consists of an opening reading frame of 1107 bp after removal of two introns. The gene encodes a protein of 369 amino acids with a predicted molecular mass of 38.3 kDa. The deduced protein sequence exhibited high homology with other fungal endopolygalacturonases. A polymerase chain reaction ( PCR ) - based strategy was used to study the expression patterns of pgg1 and pgg2 genes under different culture conditions and our results show that both genes are regulated by the carbon source at the transcriptional level. The pgg1 transcript was detected at 76 h of fungal growth in pectin while the pgg2 transcript was also induced by sucrose. The addition of yeast extract to glucose medium abolished the repressive effect of glucose, suggesting that the transcription of these genes is controlled by different mechanisms.Item Genotypic and phenotypic characterization of Staphylococcus aureus causing persistent and nonpersistent subclinical bovine intramammary infections during lactation or the dry period(Journal of Dairy Science, 2015-01) Klein, R. C.; Veh, K. A.; Ster, C.; Keefe, G.; Lacasse, P.; Scholl, D.; Roy, J. P.; Haine, D.; Dufour, S.; Talbot, v; Ribon, A. O. B.; Malouin, F.Staphylococcus aureus is a significant pathogen frequently causing persistent intramammary infections (IMI) in dairy cows. We compared some genotypic and phenotypic characteristics of 285 strains collected from quarter milk samples from cows with persistent and nonpersistent subclinical IMI across Canada. Variable number of tandem repeats typing was used to infer the persistence of the same S. aureus strain in 3 consecutive quarter milk samples collected at intervals of 3 wk during lactation or before and after dry-off. All first isolates of the series were used as the representative strains from persistent IMI and were compared with nonpersistent strains for the presence of genes seg, sen, sec, and tst as well as by spa typing. Biofilm production in vitro and hld-RNAIII expression levels were also quantified. The gene seg was associated with a reduction in the likelihood of the bacteria to cause a persistent IMI during lactation. Strains persisting through the dry period produced significantly more biofilm in vitro than strains that do not persist after calving. Also, we showed that strains expressing more hld were more likely to be non- persistent during either lactation or through the dry period. Three spa types were predominant (t529, t267, and a novel type: t13401). In the strains studied, the spa type t529 was the most frequent, and 97.0% of the strains of this spa type carried both sen and seg. Strains from the spa type t529 were less likely to cause a persis- tent IMI in the dry period. Most (86.7%) of the strains of the novel spa type (t13401) were negative for seg, sen, or both and produced significantly more biofilm in vitro than t529 and t267. The present study expanded our current knowledge on the genotypic and phenotypic traits of S. aureus strains recovered from persistent and nonpersistent IMI in Canada.Item Moving towards the immunodiagnosis of staphylococcal intramammary infections(European Journal of Clinical Microbiology & Infectious Diseases, 2014-06-20) Fabres-Klein, M. H.; Aguilar, A. P.; Silva, M. P.; Silva, D. M.; Ribon, A. O. B.Bovine mastitis is the primary disease of dairy cattle that has a great impact on the dairy industry. It is estimated that worldwide economic losses due to mastitis range between US$82 and US$131 per cow/year. A fast and efficient diagnosis of the disease remains a major bottleneck that directly influences the speed with which treatment decisions and management are undertaken. Microbiological culture remains the gold standard in the identification of bacteria that cause mastitis, but the method has inherent limitations, such as a delay in obtaining results and cost, and requires special care during the collection and processing of the sample. For this reason, multiple groups have devoted efforts to develop alternative methods that, preferably, can be easily accomplished in the field. The specificity of the antigen–antibody reaction has enabled the emergence of major diagnostic methods used in clinical practice, such as immunoassays, which have significant advantages in terms of speed, sensitivity, specificity, and portability. Commercially, immunodiagnostics have been used in the detection of various diseases in cattle. However, in several cases, only a presumptive diagnosis can be made, which requires confirmation using culture-based methods. This review discusses the immunological-based assays developed since the 1990s for the detection of Staphylococcus aureus, which is considered the primary pathogen of contagious bovine mastitis. Although no ideal antigens ensure the accurate performance of tests and the costs need to be reduced to allow for good market competitiveness, immunoassays, particularly lateral flow immunoassay and immunoagglutination, have emerged as promising tests to be used in the field.