Extended staphylococcal enterotoxin D expression in ham products.

Staphylococcal enterotoxin D (SED) is one of the most frequently recovered enterotoxins in staphylococcal food poisoning (SFP) outbreaks. The expression and production of SED were investigated in three ham products, i.e. boiled ham, smoked ham and dry-cured Serrano ham incubated at room temperature for seven days. Staphylococcus aureus was also, as a reference, grown in cultivation broth during optimal growth conditions for seven days. In boiled and smoked ham, continuous sed expression was observed throughout the incubation period with a second increase in sed expression found after five days of incubation. In smoked ham, nine times less SED per colony-forming unit of S. aureus was detected than in boiled ham. In boiled ham, the SED levels unpredictably decreased after three days of incubation. In the Serrano ham, SED was detected after five days of incubation although S. aureus growth was poor and sed expression was too low to determine. After five days of incubation, all three products contained enough SED to cause SFP. These results show that the specific production levels of SED vary in the different ham products, and that toxin production was in part uncoupled from bacterial growth.

Acetic acid increases the phage-encoded enterotoxin A expression in Staphylococcus aureus.

BACKGROUND: The effects of acetic acid, a common food preservative, on the bacteriophage-encoded enterotoxin A (SEA) expression and production in Staphylococcus aureus was investigated in pH-controlled batch cultures carried out at pH 7.0, 6.5, 6.0, 5.5, 5.0, and 4.5. Also, genomic analysis of S. aureus strains carrying sea was performed to map differences within the gene and in the temperate phage carrying sea. RESULTS: The sea expression profile was similar from pH 7.0 to 5.5, with the relative expression peaking in the transition between exponential and stationary growth phase and falling during stationary phase. The levels of sea mRNA were below the detection limit at pH 5.0 and 4.5, confirmed by very low SEA levels at these pH values. The level of relative sea expression at pH 6.0 and 5.5 were nine and four times higher, respectively, in the transitional phase than in the exponential growth phase, compared to pH 7.0 and pH 6.5, where only a slight increase in relative expression in the transitional phase was observed. Furthermore, the increase in sea expression levels at pH 6.0 and 5.5 were observed to be linked to increased intracellular sea gene copy numbers and extracellular sea-containing phage copy numbers. The extracellular SEA levels increased over time, with highest levels produced at pH 6.0 in the four growth phases investigated. Using mitomycin C, it was verified that SEA was at least partially produced as a consequence of prophage induction of the sea-phage in the three S. aureus strains tested. Finally, genetic analysis of six S. aureus strains carrying the sea gene showed specific sea phage-groups and two versions of the sea gene that may explain the different sea expression and production levels observed in this study. CONCLUSIONS: Our findings suggest that the increased sea expression in S. aureus caused by acetic acid induced the sea-encoding prophage, linking SEA production to the lifecycle of the phage.

Prolonged expression and production of Staphylococcus aureus enterotoxin A in processed pork meat.

The bacteriophage-encoded staphylococcal enterotoxin A (SEA) is the toxin most frequently reported to be involved in staphylococcal food poisoning. In this study, sea expression and SEA formation were studied in four processed pork products: boiled ham, hot-smoked ham, Serrano ham (dry-cured Spanish ham) and black pepper salami. The products were selected because of their differences in intrinsic factors. As a reference, Staphylococcus aureus was cultivated under favorable planktonic growth conditions. Expression was mainly linked to bacterial growth for both meat products and broth cultures. In liquid broth, however, the relative level of sea mRNA peaked in the late exponential phase and then rapidly declined, while in the meat products allowing immediate growth, i.e. boiled and smoked ham, active sea expression occurred throughout the incubation period of seven days. Lower levels of sea mRNA and SEA were found in smoked ham compared to boiled ham, although viable counts of S. aureus on the two products were similar. Furthermore, the SEA concentration in the boiled ham reached a maximum after three days of incubation and then unpredictably decreased. In the Serrano ham, no increase in cell number was observed until day seven, and sea expression and extracellular SEA could only be detected on days five and seven. Finally, the black pepper salami with low pH and competing microbiota proved to be a difficult environment for the survival of S. aureus. The molecular mechanism behind the behaviour of S. aureus SEA expression is discussed in connection to the life-cycle of the SEA-encoding bacteriophage and the microbial communities in these pork products.

Metrópole e expansão urbana: a persistência de processos "insustentável"

O padrão de urbanização brasileiro imprimiu às metrópoles pelo menos duas fortes características associadas ao modo predominante de "fazer cidade": apresenta componentes de "insustentabilidade" vinculados aos processos de expansão e transformação urbana e proporcionam baixa qualidade de vida a parcelas significativas da população. Esse padrão cria um espaço dual: de um lado, a cidade formal, que concentra os investimentos públicos e, de outro, seu contraponto absoluto, a cidade informal, que cresce exponencialmente na ilegalidade urbana, sem atributos de urbanidade, exacerbando as diferenças socioambientais. A transformação urbana desses espaços implica processos amplos que extrapolam as práticas correntes de regularização de parcelamentos ou urbanização de favelas (AU)