Modelling the piezo-protection effect exerted by lactate on the high pressure resistance of Listeria monocytogenes in cooked ham.

Food safety is often based on the application of several preservative (hurdle) factors whose combination must be smartly selected. The aim of the present study was to evaluate the effect of lactate and diacetate on the high pressure processing (HPP) inactivation of three L. monocytogenes strains (CTC1011, CTC1034 and Scott A) in sliced cooked ham. Inoculated vacuum-packed slices of cooked ham formulated without organic acids and with lactate, diacetate or the combination of both were pressurized at 400 MPa for different holding times and the inactivation kinetics were characterised by fitting primary and secondary models. The shape of the inactivation curves for L. monocytogenes depended on both product formulation and strain. Interestingly, lactate caused a dose-dependent piezo-protection in all three strains, as the HPP inactivation rate decreased in cooked ham formulated with increasing amounts of lactate and in comparison with the control product. The design, validation and implementation of HPP requires a tailor-made approach, considering product formulation and selection of strain/s.

Antibiotic resistance of lactobacilli isolated from two italian hard cheeses.

One hundred forty-one lactobacilli strains isolated from natural whey starter cultures and ripened Grana Padano and Parmigiano Reggiano cheeses were tested for their susceptibility to 13 antibiotics, in particular, penicillin G, ampicillin, amoxicillin, oxacillin, cephalotin, cefuroxime, vancomycin, gentamicin, tetracycline, erythromycin, clindamycin, co-trimoxazole, and nitrofurantoin. The strains belonged to the species Lactobacillus helveticus, L. delbrueckii subsp. lactis, L. rhamnosus, and L. casei. The strains of the first two species were isolated from whey starter cultures, and the strains of the last two species were from the ripened cheeses. Significant differences among the strains in their antibiotic resistance were found in relation to the type of cheese and, especially, the strains from Parmigiano Reggiano were more resistant to gentamicin and penicillin G. The strains isolated in the ripened cheese were generally more resistant than those isolated from natural whey starter cultures; in particular, significant differences regarding oxacillin, vancomycin, cephalotin, and co-trimoxazole were observed. Finally, no significant difference in relation to the type of cheese was found among the thermophilic lactobacilli isolated from whey cultures, while the facultatively heterofermentative lactobacilli isolated from Parmigiano Reggiano showed higher resistance toward gentamicin and penicillin G than did the same species isolated from Grana Padano.

Modeling the aminogenic potential of Enterococcus faecalis EF37 in dry fermented sausages through chemical and molecular approaches.

Amino acid decarboxylase activity of the Enterococcus faecalis strain EF37 was monitored during fermentation and ripening of a traditional dry fermented sausage from Northern Italy (Salame Veronese) by means of microbiological, chemical, and molecular approaches in relation to three technological factors: fermentation temperature, sodium chloride concentration, and amount of glucose added to the meat mixture. Besides the analytical determination of tyramine and phenylethylamine accumulation and the counts of enterococci, the presence and quantification of the tyrosine decarboxylase gene (tdc) and its mRNA transcript were also investigated by using real-time PCR. According to the mathematical models obtained, all of the three factors studied were statistically significant and microbiologically relevant for the early development of enterococci, although the fermentation temperature had a more relevant influence on the enterococcal viable cells of the ripened product. Sodium chloride concentration was the most determinant factor of the final tyramine and 2-phenylethylamine accumulation and also of the levels of tdc present in the final product. In contrast, an effect of glucose concentration on tdc expression was observed in the last period of ripening. Moreover, increasing amounts of sodium chloride and decreasing fermentation temperature resulted in a reduced tdc expression. This is the first time that bacterial tyrosine decarboxylase potential is directly examined through a molecular approach in a fermented meat. The quantification of tdc and its transcript can help to elucidate the critical steps and factors during food manufacturing at which bacterial aminogenesis is possible, thus allowing researchers to propose technological measures to control decarboxylase activities.

Effects of sub-lethal concentrations of hexanal and 2-(E)-hexenal on membrane fatty acid composition and volatile compounds of Listeria monocytogenes, Staphylococcus aureus, Salmonella enteritidis and Escherichia coli.

In recent years, the interest in the possible use of natural alternatives to food additives to prevent bacterial and fungal growth has notably increased. Plants and plant products can represent a source of natural alternatives to improve the shelf-life and the safety of food. Some of these compounds, i.e. hexanal, hexanol, 2-(E)-hexenal and 3-(Z)-hexenol, produced throughout the lipoxygenase pathway have important roles in plant defence with a protective action towards microbial proliferation in wounded areas. Otherwise, hexanal and 2-(E)-hexenal have evidenced a marked antimicrobial activity against food spoilage and pathogenic microbial species both in model and real systems. The precise mechanisms of action of all these antimicrobial compounds are not yet clear. Because the usage of these compounds as antimicrobials in foods has to be supported by the comprehension of mechanisms of action of these compounds, the overall purpose of this work was to study the modifications of the cell membrane and volatile compounds of Listeria monocytogenes, Staphylococcus aureus, Salmonella enteritidis and Escherichia coli during the growth in the presence of sub-lethal doses of these two aldehydes. The results obtained evidenced that the tested molecules induced noticeable modifications of the composition of cell membrane and the volatile compounds produced during the growth. Although specific differences in relation to the species considered were identified, 2-(E)-hexenal and hexanal induced a marked increase of some membrane associated fatty acids, both linear and branched fatty acids as well as unsaturated fatty acids, and released free fatty acids.

Rapid detection and quantification of tyrosine decarboxylase gene (tdc) and its expression in gram-positive bacteria associated with fermented foods using PCR-based methods.

In this study, PCR-based procedures were developed to detect the occurrence and quantify the expression of the tyrosine decarboxylase gene (tdc) in gram-positive bacteria associated with fermented foods. Consensus primers were used in conventional and reverse transcription PCR to analyze a collection of 87 pure cultures of lactic acid bacteria and staphylococci. All enterococci, Staphylococcus epidermidis, Lactobacillus brevis, Lactobacillus curvatus, and Lactobacillus fermentum strains and 1 of 10 Staphylococcus xylosus strains produced amplification products with the primers DEC5 and DEC3 in accordance with results of the screening plate method and with previously reported result obtained with high-performance liquid chromatography. No amplicons were obtained for tyramine-negative strains, confirming the high specificity of these new primers. A novel quantitative real-time PCR assay was successfully applied to quantify tdc and its transcript in pure cultures and in meat and meat products. This assay allowed estimation of the influence of different variables (pH, temperature, and NaCl concentration) on the tdc expression of the tyraminogenic strain Enterococcus faecalis EF37 after 72 h of growth in M17 medium. Data obtained suggest that stressful conditions could induce greater tyrosine decarboxylase activity. The culture-independent PCR procedures developed here may be used for reliable and fast detection and quantification of bacterial tyraminogenic activity without the limitations of conventional techniques.

Assessment of safety, nutritional, and spoilage characteristics of different lagoon grey mullets (Liza ramada, Liza aurata, and Liza saliens).

Different lagoon grey mullets such as Liza ramada (thinlip mullet), Liza aurata (golden grey mullet), and Liza saliens (leaping grey mullet) were analyzed for their nutritional, microbiological, and safety parameters. The microbiological values never exceeded the lower limits stipulated by the Italian Higher Institute of Health. The pathogenic species frequently associated with seafood (Salmonella, Listeria monocytogenes, Vibrio cholerae, Vibrio parahaemolyticus, and Aeromonas hydrophila) were never detected. The absence of coliforms and of Escherichia coli was noted in all fish species after 4 days of storage in ice. Heavy metals such as cadmium and mercury were always below the detection limits (0.01 mg/kg). All three fish species had low levels of total biogenic amines (80 to 100 mg/kg), and the presence of histamine was sporadic. All Liza species, particularly L. ramada and L. saliens, are a good source of omega3 long-chain polyunsaturated fatty acids.

Role of surface-inoculated Debaryomyces hansenii and Yarrowia lipolytica strains in dried fermented sausage manufacture. Part 2: Evaluation of their effects on sensory quality and biogenic amine content.

The aim was to study the effects of Debaryomyces hansenii and Yarrowia lipolytica strains, used with lactic acid starter cultures (Lactobacillus plantarum), in the manufacture of dried fermented sausages in order to understand their effects on volatile profile, biogenic amine content and sensory properties. The experimental data showed that every yeast strain produced a specific profile of volatile metabolic products. The yeasts also gave sausages with distinctive sensory properties. The degree of mincing also influenced these properties, but none of these factors had significant influence upon the accumulation of biogenic amines.

Evaluation of the antimicrobial activity of citrus essences on Saccharomyces cerevisiae.

The aim of this research was to assess the antimicrobial activity of nine different industrial essences used in a soft drink factory in relation to their composition, as well as to verify the role of vapor pressure on their bioactivity. The essences were tested against a Saccharomyces cerevisiae strain isolated from spoiled soft drinks. The tests were carried out by adding the essences directly to a liquid medium or into the headspace of closed systems inoculated with the yeast. The headspace composition was evaluated through a solid phase microextraction-gas chromatography technique. The use of a mass spectrometer allowed the identification of the peaks detected. The microbial growth was indirectly monitored by measuring the metabolic CO2 released by the yeast. The results obtained indicated that the most effective essences were characterized by the highest concentration of some terpenes, such as citral, beta-pinene, and p-cymene. Moreover, all of the essences were more bioactive when added directly to the liquid medium.

Application of hexanal, E-2-hexenal, and hexyl acetate to improve the safety of fresh-sliced apples.

The aims of this work were to evaluate the effects of different concentrations of hexanal, (E)-2-hexenal, hexyl acetate, and their mixtures on the fate of pathogenic species such as Escherichia coli, Salmonella enteritidis, and Listeria monocytogenes inoculated in model systems as well as the antimicrobial activity against the target species of the chosen molecules when added to the packaging atmosphere of inoculated fresh-sliced apples. The result obtained in this work pointed out the potential use of compounds such as hexanal, (E)-2-hexenal, and hexyl acetate for both the extension of shelf life and an improvement of hygienic safety of "minimally processed foods". In fact, hexanal, (E)-2-hexenal, and hexyl acetate had a significant inhibitory effect against pathogen microorganisms frequently isolated from raw materials (E. coli, S. enteritidis, and L. monocytogenes) when inoculated in both model and real systems. In this last condition, these compounds, at the levels used (150, 150, and 20 ppm for hexanal, hexyl acetate, and (E)-2-hexenal, respectively), displayed a bactericide effect on L. monocytogenes and they exhibited significant extensions of lag phase of E. coli and S. enteritidis inoculated at levels of 10(4)-10(5) CFU/g.

Effects of carvacrol, (E)-2-hexenal, and citral on the thermal death kinetics of Listeria monocytogenes.

Carvacrol, (E)-2-hexenal, and citral at sublethal concentrations combined with isothermal heating between 55 and 68°C were assessed for their effects on Listeria monocytogenes 56LY. Experimental survival curves were obtained and fitted to the Weibull equation to estimate parameters describing their shape and rate. These parameters were further used to assess the impact of this combination of treatments on the cell resistance distribution during inactivation. The sublethal concentrations of the aroma compounds used (i.e., 50 mg/liter citral, 65 mg/liter (E)-2-hexenal, and 30 mg/liter carvacrol) did not prevent the growth of L. monocytogenes at 37°C but did enhance inactivation. Between 55 and 63°C, the presence of the aroma compounds reduced by about two-thirds the time needed for a 5-log reduction of the microbial counts, e.g., from 145.75 h in the control treatment (at 55°C) to 40.84 h in the presence of carvacrol (at the same temperature). The mean and variance observed in the frequency distribution of resistance were reduced as the temperature increased. The results obtained at isothermal temperatures and with single aroma components provide basic information regarding components frequently found in essential oils, which can be used in combination with less extreme thermal treatments to provide energy conservation and improve food quality.

Modeling of combined effects of citral, linalool and beta-pinene used against Saccharomyces cerevisiae in citrus-based beverages subjected to a mild heat treatment.

The aim of this work was to evaluate the antimicrobial activity of three terpenes (citral, linalool and beta-pinene), in combination with a mild heat treatment (55 degrees C, 15 min). The study has been carried out on an orange based soft drink inoculated using a wild strain of Saccharomyces cerevisiae. The results, expressed as growth/no-growth data, were analyzed with the logistic regression. A model comprising only of significant individual parameters (p < or = 0.05) and describing the relationships between terpene concentrations and the probability of having stable beverages was obtained. When citral and beta-pinene were combined, the citral concentration required to achieve a 50% probability of having stable bottles (P=0.5) dropped from 100.9 microL/L in the absence of beta-pinene to 49.3 microL/L in the presence of 20 microL/L of beta-pinene. The mixture of citral and linalool was less effective, in fact, the same probability (P=0.5) was obtained combining 60 microL/L of linalool with 35.1 microL/L of citral. The addition of 20 microL/L of linalool and beta-pinene reinforced citral bioactivity and the concentration of citral needed to reach P=0.5 fell from 100.9 microL/L in the presence of citral alone to 42.0 microL/L. The presence of both linalool and beta-pinene at a concentration of 40 or 60 microL/L in the absence of citral led to a lower spoilage probability (P=0.58 and P=0.93, respectively). It can be concluded that the antimicrobial potential of the three terpenes alone can be strengthened combining appropriate concentrations of each of them. This study confirmed also the potentiating effect of a mild temperature treatment on the antimicrobial efficacy of the molecules. Neither the thermal treatment alone nor the presence of the terpenes at their maximum concentrations (without thermal treatment) were able to guarantee the microbial stability of the beverages.

Antimicrobial activity of aroma compounds against Saccharomyces cerevisiae and improvement of microbiological stability of soft drinks as assessed by logistic regression.

The combined effects of a mild heat treatment (55 degrees C) and the presence of three aroma compounds [citron essential oil, citral, and (E)-2-hexenal] on the spoilage of noncarbonated beverages inoculated with different amounts of a Saccharomyces cerevisiae strain were evaluated. The results, expressed as growth/no growth, were elaborated using a logistic regression in order to assess the probability of beverage spoilage as a function of thermal treatment length, concentration of flavoring agents, and yeast inoculum. The logit models obtained for the three substances were extremely precise. The thermal treatment alone, even if prolonged for 20 min, was not able to prevent yeast growth. However, the presence of increasing concentrations of aroma compounds improved the stability of the products. The inhibiting effect of the compounds was enhanced by a prolonged thermal treatment. In fact, it influenced the vapor pressure of the molecules, which can easily interact within microbial membranes when they are in gaseous form. (E)-2-Hexenal showed a threshold level, related to initial inoculum and thermal treatment length, over which yeast growth was rapidly inhibited. Concentrations over 100 ppm of citral and thermal treatment longer than 16 min allowed a 90% probability of stability for bottles inoculated with 10(5) CFU/bottle. Citron gave the most interesting responses: beverages with 500 ppm of essential oil needed only 3 min of treatment to prevent yeast growth. In this framework, the logistic regression proved to be an important tool to study alternative hurdle strategies for the stabilization of noncarbonated beverages.