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The nucleotide sequence of the hmc operon from Desulfovibrio vulgaris subsp. vulgaris Hildenborough indicated the presence of eight open reading frames, encoding proteins Orfl to Orf6, Rrfl, and Rrf. Orfi is the periplasmic, high-molecular-weight cytochrome (Hmc) containing 16 c-type hemes and described before

A standardized disinfectant test for Staphylococcus aureus cells in biofilms was developed. Two disinfectants, the membrane-active compound benzalkonium chloride (BAC) and the oxidizing agent sodium hypochlorite, were used to evaluate the biofilm test. S. aureus formed biofilms on glass, stainless steel, and polystyrene in a simple system with constant nutrient flow that mimicked as closely as possible the conditions used in the current standard European disinfectant test (EN 1040). The biofilm that was formed on glass contained cell clumps and extracellular polysaccharides. The average surface coverage was 60%, and most (92%) of the biofilm cells were viable. Biofilm formation and biofilm disinfection in different experiments were reproducible. For biofilms exposed to BAC and hypochlorite the concentrations needed to achieve 4-log killing were 50 and 600 times higher, respectively, than the concentrations needed to achieve this level of killing with the European phase 1 suspension test cells. Our results show that a standardized disinfectant test for biofilm cells is a useful addition to the current standard tests.

Quantitative microbiological models predicting proliferation of microorganisms relevant for food safety and/or food stability are useful tools to limit the need for generation of biological data through challenge testing and shelf-life testing. The use of these models requires quick and reliable methods for the generation of growth data and estimation of growth parameters. Growth parameter estimation can be achieved using methods based on plate counting and methods based on measuring the optical density. This research compares the plate count method with two optical density methods, namely, the 2-fold dilution (2FD) method and the relative rate to detection (RRD) method. For model organism Bacillus cereus F4810/72, the plate count method and both optical density methods gave comparable estimates for key growth parameters. Values for the maximum specific growth rate ( max ) derived by the 2FD method and by the RRD method were of the same order of magnitude, but some marked differences between the two approaches were apparent. Whereas the 2FD method allowed the derivation of values for lag time () from the data, this was not possible with the RRD method. However, the RRD method gave many more data points per experiment and also gave more data points close to the growth boundary. This research shows that all three proposed methods can be used for parameter estimation but that the choice of method depends on the objectives of the research.

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Milk quality is assessed using bulk milk analysis and by farm audits in the Netherlands. However, the extent of the effect that dairy farm audits have on milk quality is unknown. Data from over 13,000 audits performed on 12,855 dairy farms from February 2006 to April 2008 were merged with laboratory test results of 325,150 bulk milk samples collected 6 mo before and after the audit. A linear mixed model with the method of restricted maximum likelihood was conducted to study whether the total bacterial counts (TBC) of bulk milk were lower during the periods before and after the dairy farm audit. Results showed that TBC values were 2 to 6% lower (i.e., 0.010 to 0.026 log cfu/mL) for a period from 1.5 to at least 6 mo after an audit. Additionally, several variables were significantly associated with bulk milk TBC values: seasonality, total number of attention points (given if some checklist points were not appropriate), audit type, audit result, and the categories milking equipment maintenance, and utility room-tank maintenance. The TBC values increased with a higher level of attention points. Furthermore, the farms rejected based on the audit results had the highest average TBC values and the approved farms had the lowest values. If dairy farms had an overall negative audit assessment and consequently needed a re-audit in the following year, the TBC values of bulk milk were more likely to be higher. Auditing may provide dairy farmers the opportunity to receive advice about factors that influence bulk milk TBC values, for a period of at least 6 mo following the audit.

Enterobacter sakazakii can be present, although in low levels, in dry powdered infant formulae, and it has been linked to cases of meningitis in neonates, especially those born prematurely. In order to prevent illness, product contamination at manufacture and during preparation, as well as growth after reconstitution, must be minimized by appropriate control measures. In this publication, several determinants of the growth of E. sakazakii in reconstituted infant formula are reported. The following key growth parameters were determined: lag time, specific growth rate, and maximum population density. Cells were harvested at different phases of growth and spiked into powdered infant formula. After reconstitution in sterile water, E. sakazakii was able to grow at temperatures between 8 and 47°C. The estimated optimal growth temperature was 39.4°C, whereas the optimal specific growth rate was 2.31 h ؊1 . The effect of temperature on the specific growth rate was described with two secondary growth models. The resulting minimum and maximum temperatures estimated with the secondary Rosso equation were 3.6°C and 47.6°C, respectively. The estimated lag time varied from 83.3 ؎ 18.7 h at 10°C to 1.73 ؎ 0.43 h at 37°C and could be described with the hyperbolic model and reciprocal square root relation. Cells harvested at different phases of growth did not exhibit significant differences in either specific growth rate or lag time. Strains did not have different lag times, and lag times were short given that the cells had spent several (3 to 10) days in dry powdered infant formula. The growth rates and lag times at various temperatures obtained in this study may help in calculations of the period for which reconstituted infant formula can be stored at a specific temperature without detrimental impact on health.Enterobacter sakazakii is a motile, peritrichous, gram-negative rod that occasionally causes neonatal meningitis and sepsis, with mortality rates of 40 to 80% (3). The recovery of E. sakazakii from samples of commercially available dry powdered infant formulae has been reported (4,8,9). E. sakazakii organisms in infant formula have been associated with outbreaks of meningitis, sepsis, and necrotizing enterocolitis in premature and full-term infants, particularly those with predisposing medical conditions (17). Although the levels of E. sakazakii occurring in dry powdered infant formula are generally very low, reconstituted infant formula is a good medium for growth. When present in dry formula, E. sakazakii may grow during preparation, cooling, storage, and holding of the bottles, increasing the probability of illness. Occasional contamination of dried infant formula during manufacture is a source of the microorganism's occurrence in reconstituted product. However, as E. sakazakii has been detected in various other dry environments (7), contamination may also occur during reconstitution of dried infant formula in hospitals or at home.In order to prevent illness, product contamination at manufacture and/or during preparat...