This study aimed to isolate and characterize a microbial culture able to degrade sulfonamides. Sulfamethoxazole (SMX)-degrading microorganisms were enriched from activated sludge and wastewater. The resultant mixed culture was composed of four bacterial strains, out of which only Achromobacter denitrificans PR1 could degrade SMX. This sulfonamide was used as sole source of carbon, nitrogen and energy with stoichiometric accumulation of 3-amino-5-methylisoxazole. Strain PR1 was able to remove SMX at a rate of 73.6 ± 9.6 µmolSMX/gcell dry weight h. This rate more than doubled when a supplement of amino acids or the other members of the mixed culture were added. Besides SMX, strain PR1 was able to degrade other sulfonamides with anti-microbial activity. Other environmental Achromobacter spp. could not degrade SMX, suggesting that this property is not broadly distributed in members of this genus. Further studies are needed to shed additional light on the genetics and enzymology of this process.

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Aqueous extracts obtained from cell suspension cultures of Centaurea calcitrapa were used as proteolytic additive in the manufacture of a commercial bovine cheese, coagulated with animal rennet and typically ripened for 28 d. The cheese was assessed in comparison to standard cheese for two levels of addition of said extract, viz. 0.61 and 1.22 mg of total protein mL\. The qualitative and quantitative evolutions of the nitrogen fractions were monitored in the experimental cheeses throughout the whole ripening period. In general, the chemical compositions of the cheeses were di!erent depending on the amount of extract used, but no signi"cant di!erences could be detected in the ripening index. With regard to electrophoretic pro"les, the two types of cheese could be distinguished until up to ca. 7 d of ripening, but di!erences did essentially vanish by 28 d.

This work was aimed at enumerating the viable microorganisms in ripened Serra da Estrela cheeses, manufactured from both refrigerated and non-refrigerated milk, in various dairies located throughout the demarcated region. Scanning electron microscopy was used to analyze the microstructure, and thus aid in understanding possible differences in their microbiological profile. The cheeses were allowed to ripen under controlled conditions, and sampled at 60, 90, 120, 150 and 180 d following manufacture. Viable numbers of lactic acid bacteria, staphylococci, Enterobacteriaceae and yeasts were obtained following standard plate counting on a number of selective media. Lactococcus was the most abundant genus (above 10 8 cfu g À1 of cheese) up to 120 d of ripening. No significant microstructural differences were observed in cheeses manufactured in different dairies over the ripening process. However, microstructural differences were apparent between cheeses manufactured with refrigerated versus non-refrigerated milk. r