Antonietta Galli

Microbiological Quality and Shelf Life Modeling of Ready-to-Eat Cicorino

Growth of the microbial population resident in ready-to-use fresh cut cicorino, a variety of Chichorium intybus, was determined in the various preparation steps with the aim of defining the hazard critical control points. The investigation concerned cut cicorino from two producers. During the process a 1- to 1.5-fold increase of microbial counts was observed, and the retail product showed values of 10(5) to 10(6) CFU/g. The shelf life of the product was kinetically modeled in order to check the effects of storage temperature and assess the microbial indexes most relevant for hygiene and quality of the distributed product. A modified Gompertz function described the kinetics of microbial growth and allowed definition of a stability time and its dependence on temperature. Stability times were of 0.3, 3.7, and 4.7 days at storage temperatures of 20, 10, and 5 degrees C, respectively. Q10 (the fold decrease of stability time for an increase of 10 degrees C) was 3.85. The results from this study may be used to predict the effects of temperatures experienced in the distribution chain on bacterial levels in cicorino.

Bactericidal activity of chlorine dioxide against Escherichia coli in water and on hard surfaces.

The efficacy of chlorine dioxide as a disinfectant was evaluated against cells of Escherichia coli ATCC 11229 in aqueous suspension and adhering to the surfaces of stainless steel AISI 304 and PVC. The concentrations tested ranged from 0.7 to 14 mg/liter; the exposure times investigated were 30 s and 1, 2, 4, and 8 min. When the bacteria were suspended in water with 1.4 mg/liter of chlorine dioxide, a 10(5)-fold reduction of the initial viable count occurred within 30 s; when cells were attached to the steel surface, the same rate of inactivation took place only after 6 min with 7 mg/liter or 4 min with 14 mg/liter of chlorine dioxide. A 5-log reduction was not obtained when organisms were adhered to polyvinyl chloride (PVC). Scanning electron microscope micrographs of contaminated surfaces revealed that the PVC was very rough with pores much larger in diameter than the cells. Time values determining a 90% reduction of the E. coli population (90% killing time) were calculated for each concentration of disinfectant tested in suspension and on the steel surface. If the same experimental conditions were strictly adopted, linear functions of the log of bacterial inactivation could be plotted (log 90% killing time versus log concentration of disinfectant). This work showed that results obtained with suspension tests could not be used to estimate disinfection of hard surfaces.

Survival and residual activity of Lactobacillus acidophilus frozen cultures under different conditions

Eight strains of the Lactobacillus acidophilus complex were subjected, as concentrated cultures in skim milk, to different freezing treatments (rapidly in liquid nitrogen and more slowly in cold air at - 30 °C), then stored at - 80 and - 30 °C for 9 months. Survival was determined by plate counting with and without bile salts or sodium chloride at the highest tolerated concentration for each strain, to distinguish the undamaged population from the total population. Fermentative activity was measured as total lactic acid production by thawed cultures under standard conditions. Higher survival rates and greater activity were always obtained by storing cultures at -80 °C, but most strains stored at -30 °C also survived well. Analysis of variance revealed that the viability of the frozen cultures was affected more by storage temperature than by cooling rate. Selective media were unable to distinguish the active population from the total surviving population. The correlation between values for activity and survival with selective media was poor.

Phenotypic and genotypic characterization of Enterococcus spp. of different origins.

Sixty-four strains of Enterococcus spp. of different origin were identified using traditional phenotypic, biochemical and cultural tests, together with molecular tests. API 20 STREP tests identified the species at a preliminary level, only one strain remaining unidentified. Strains belonging to the genus Enterococcus were tested with genus-specific primers, while species-level identification was carried out with the 16S–23S rDNA intergenic region (ITS) and species-specific primers for E. faecium and E. faecalis. Those strains found to be negative with the species-specific primers were subjected to 16S rDNA partial sequencing.

Production of cyclopiazonic acid by molds isolated from Taleggio cheese.

Twenty-seven strains of Penicillium were isolated from the rind of Taleggio, a typical Italian cheese, so that we could test their capacity to produce cyclopiazonic acid (CPA); all strains produced CPA. The production was strongly influenced by the strain variety and growth conditions. Strains incubated at 25 degrees C for 7 days always produced CPA in mannitol broth, with concentrations ranging from 0.02 to 1 microg/ml, whereas only 33% of strains grown in yeast-extract broth produced CPA, with a maximum value of 0.1 microg/ml. In milk, maximum production (1.6 microg/ml) was observed after 14 days of incubation at 25 degrees C. In order to evaluate the presence of the toxin and its capacity for migrating into the cheeses, the rind, the cheese near the rind, and the cores from six Taleggio cheeses were analyzed. CPA was present in five cheeses, with a maximum concentration of 0.25 mg/kg in one rind, and in one cheese, the toxin migrated to the core. A positive correlation between CPA production and surface mold was found.

Evidence for enantiomorphic-enantiotopic group discrimination in diol dehydratase-catalyzed dehydration of meso-2,3-butanediol

Abstract The conversion of meso -2,3-butanediol ( 1 ) into 2-butanol ( 3 ) by Lactobacillus brevis, via diol dehydratase-catalyzed reaction to 2-butanone ( 2 ), was shown to occur with complete discrimination between the two enantiomorphic-enantiotopic 1-hydroxyethyl groups of 1 .

STEREOCHEMISTRY AND FATE OF HYDROGEN ATOMS IN THE DIOL-DEHYDRATASE-CATALYZED DEHYDRATION OF MESO-BUTANE-2,3-DIOL

The transformation of meso-butane-2,3-diol into butan-2-ol by a strain of Lactobacillus brevis occurs through a diol-dehydratase-catalyzed conversion of the diol into butan-2-one, which is then reduced to the secondary alcohol by dehydrogenases. Experiments performed with deuterated meso-butane-2,3-diols showed that the dehydration reaction brings about an inversion of configuration at the (R)-configured C-atom of meso-butane-2,3-diol as a consequence of the substitution of the OH group by a H-atom; at the same time, the H-atom already bound to the (R) C-atom is retained in the resulting methylene group. The H-atom replacing the OH group was assessed to come from the medium since the H-atom at the (S)-configured C-atom was completely lost. By contrast, in the case of the conversion of (RS)-propane-1,2-diol into propan-1-ol under the same fermentation conditions, an extensive H-transfer (ca. 80%) from the primary-alcohol function to the adjacent C-atom was observed. This fact is taken as an indication of different modes in which the two substrates are processed by the enzyme (despite the same stereochemical outcome). A speculative hypothesis is presented to interpret such a dissimilarity.