Prodotti della tradizione e contenuto di amine biogene alternative alla Low tyramine diet per la sostenibilità dei prodotti di nicchia e la salubrità del consumatore

Biogenic amines (BA) are present in a wide range of foods and mainly can be produced in high amounts by microorganisms through the activity of amino acid decarboxylases. Excessive consumption of foods with large concentrations of these compounds can induce adverse reactions such as nausea, headaches, rashes and changes in blood pressure. These problems are more severe in consumers with less efficient detoxification systems because of their genetic constitution or their medical treatments. The most common and powerful BA found in foods are histamine, tyramine and putrescine. Actually, there is no specific legislation regarding BA content in many fermented products, except for histamine; it is the only BA for which maximum levels in fish products have been set. So, it is generally assumed that these compounds should not be allowed to accumulate. Many factors such as bacterial density, synergistic effects between microorganisms, level of proteolysis (availability of substrate), pH, salt, use of starter cultures, sanitization procedures adopted and conditions and time of ripening process are found to have limiting effects on the build-up of amines. Moreover, improved knowledge of the factors involved in the synthesis and accumulation of BA should lead to reduce in their incidence in foods

A Survey of Antibiotic Resistance in Micrococcaceae Isolated from Italian Dry Fermented Sausages

The transfer of bacteria that are resistant to antimicrobial agents or resistance genes from animals to humans via the food chain is increasingly a problem. Therefore, it is important to determine the species and the numbers of bacteria involved in this phenomenon. For this purpose, 148 strains of microstaphylococci were isolated from three types of Italian dry fermented sausages. Eight of 148 strains belonged to the genera Kocuria and Micrococcus. The remaining 140 strains belonged to 11 different species of the genus Staphylococcus. The species most frequently isolated was Staphylococcus xylosus, followed by Staphylococcus saprophyticus and Staphylococcus aureus. Antibiotic resistance levels differed among the species and depended on the strain origin. Microstaphylococci were generally susceptible to beta-lactams, but 12 strains were resistant to methicillin, 8 were resistant to oxacillin, and 9 were resistant to penicillin G. No resistance was observed for aminoglicosides and cephalosporines. Many strains were resistant to sulfonamide, colistin suphate, tetracyclin, and bacitracin. Two strains of S. aureus, four strains of S. xylosus, and one strain of Staphylococcus sciuri were able to grow in the presence of 8 microg of vancomycin per g, but all strains were susceptible to teicoplanin. Twenty-two microstaphylococci were resistant to at least five of the tested antibiotics. The multiresistant strain S. aureus 899 was unaffected by eight antibiotics, including vancomycin and methicillin, indicating that a more prudent use of antibiotics in animal husbandry and better hygienic conditions during production should be encouraged because they can play a major role in reducing the incidence of such multiresistant microorganisms and the possible spread of the genetic elements of their resistance

Biogenic Amines in Raw and Processed Seafood

The presence of biogenic amines (BAs) in raw and processed seafood, associated with either time/temperature conditions or food technologies is discussed in the present paper from a safety and prevention point of view. In particular, storage temperature, handling practices, presence of microbial populations with decarboxylase activity and availability of free amino acids are considered the most important factors affecting the production of BAs in raw seafood. On the other hand, some food technological treatments such as salting, ripening, fermentation, or marination can increase the levels of BAs in processed seafood. The consumption of high amount of BAs, above all histamine, can result in food borne poisoning which is a worldwide problem. The European Regulation established as maximum limits for histamine, in fishery products from fish species associated with high histidine amounts, values ranging from 100 to 200 mg/kg, while for products which have undergone enzyme maturation treatment in brine, the aforementioned limits rise to 200 and 400 mg/kg. Preventive measures and emerging methods aiming at controlling the production of BAs are also reported for potential application in seafood industries

Biogenic Amines in Italian Pecorino Cheese

The quality of distinctive artisanal cheeses is closely associated with the territory of production and its traditions. Pedoclimatic characteristics, genetic autochthonous variations, and anthropic components create an environment so specific that it would be extremely difficult to reproduce elsewhere. Pecorino cheese is included in this sector of the market and is widely diffused in Italy (∼62.000t of production in 2010). Pecorino is a common name given to indicate Italian cheeses made exclusively from pure ewes’ milk characterized by a high content of fat matter and it is mainly produced in the middle and south of Italy by traditional procedures from raw or pasteurized milk. The microbiota plays a major role in the development of the organoleptic characteristics of the cheese but it can also be responsible for the accumulation of undesirable substances, such as biogenic amines (BA). Bacterial amino acid decarboxylase activity and BA content have to be investigated within the complex microbial community of raw milk cheese for different cheese technologies. The results emphasize the necessity of controlling the indigenous bacterial population responsible for high production of BA and the use of competitive adjunct cultures could be suggested. Several factors can contribute to the qualitative and quantitative profiles of BA’s in Pecorino cheese such as environmental hygienic conditions, pH, salt concentration, water activity, fat content, pasteurization of milk, decarboxylase microorganisms, starter cultures, temperature and time of ripening, storage, part of the cheese (core, edge), and the presence of cofactor (pyridoxal phosphate, availability of aminases and deaminases). In fact physico-chemical parameters seem to favor biogenic amine-positive microbiota; both of these environmental factors can easily be modulated, in order to control growth of undesirable microorganisms. Generally, the total content of BA’s in Pecorino cheeses can range from about 100–2400 mg/kg, with a prevalence of toxicologically important BA’s, tyramine and histamine. The presence of BA is becoming increasingly important to consumers and cheese-maker alike, due to the potential threats of toxicity to humans and consequent trade implications

In silico RFLP Analysis of 16S rRNA Genes: A Helpful Application for Distinguishing Bifidobacteria from Human and Animal Source

Bifidobacterial species are widespread in gastrointestinal tracts of mammalian and other animals; they can be found in extra body environment only after a fecal contamination or human intentional addition (as the case of probiotics). Interestingly their occurrence is strictly linked to their hosts with a clear demarcation between animal and human species. PCR-restriction fragment length polymorphism (PCR-RFLP) on the 16S rRNA gene, using Alul, and TaqI restriction enzymes, have been utilized to distinguish the animal or human source of 64 strains belonging to 13 Bifidobacterium species (Delcenserie et al. [15]). Our aim was to test this method updating an in silico restriction analysis on the available 16S rRNA gene sequences of all 55 currently described taxa of Bifidobacterium genus. Our results confirmed the reliability of this method, optimized with the use of three restriction enzymes: Alul, TaqI and MaeIII, as a fast and simple strategy to determine the origin (human or animal) of bifidobacteria. Interestingly, the bifidobacterial species recently isolated from non-human primates cluster in the group of animal source except the bifidobacterial species isolated from higher non-human primates closest to humans such as apes (chimpanzee, orangutan and gorilla) that clusters with human group. Moreover, B. minimum, B. subtile and B. mongoliense isolated only from extrabody environment of which the source is unknown clustered with animal species. The in silico RFLP-PCR confirmed its powerful ability to attribute the primary source of occurrence (human or animal) for bifidobacterial species to the human or animal habitat

A survey of Saccharomyces populations associated with wine fermentations from the Apulia region (South Italy)

The aim of this paper was to investigate the genetic and phenotypic characteristics of yeasts isolated from samples of grape musts collected from four different areas of Apulia region. The 68 yeast isolates were identified asSaccharomyces cerevisiae by PCR-RFLP of 5.8S-ITS region of the rRNA gene. Individual isolates were differentiated by RAPD-PCR and AFLP. The following oenological traits were studied: fermentation power, resistance to cycloheximide, alcohol and SO2, formation of SO2 and H2S, β-glucosidase activity, and production of biogenic amines and secondary compounds. Many phenotypes were common to several yeasts isolated from the four different areas, such as high SO2 resistance and fermentation power. In addition, someS. cerevisiae isolates showed a β-glucosidase activity and others had a high resistance to cycloheximide. All the strains formed biogenic amines. Solid Phase Microextraction was used to determine secondary compounds produced in wine by the single yeast cultures

Influence of organic viticulture on non- Saccharomyces wine yeast populations

This study evaluated the population dynamics of non-Saccharomyces biota during spontaneous fermentation of organic musts. Non-Saccharomyces yeasts were found to be present at high levels during all fermentations. A total of 543 yeast colonies were isolated, 190 from Lysine-Medium (LM) agar, 254 from Wallerstein Laboratory Nutrient (WLN) agar and 99 from YPD agar. To estimate yeast population dynamics during spontaneous fermentation a genotypic approach was applied. PCR-RFLP of the ITS1-5.8S rDNA-ITS2 region and sequence determination of the D1/D2 region of the 26S rRNA gene enabled identification of the yeast isolates at the species level. Hanseniaspora uvarum, Metschnikowia fructicola and Candida zemplinina predominated, while Issatchenkia terricola, Issatchenkia orientalis and Pichia sp. were identified with a lower frequency. Hanseniaspora uvarum, M. fructicola and C. zemplinina represented 43%, 31% and 11% of the total non-Saccharomyces population isolated, respectively. Some yeast isolates were shown to be closely related to Hanseniaspora spp. and Candida spp. on the basis of the D1/D2 sequences. Based on those results, the coexistence of different Hanseniaspora and Pichia species in grape musts was supposed, and their complete identification was achieved using additional molecular markers. Moreover, strain typing and differentiation was carried out by RAPD-PCR. High strain polymorphisms were observed in the different species. For some strains, appreciable properties were demonstrated both in vitro by the API-ZYM test and in must. In must microvinification some strains showed good fermentation performances, low production of acetic acid and a partial capability to degrade malic acid

Multistarter from Organic Viticulture for Red Wine Montepulciano d’Abruzzo Production

In the last years the use of a multistarter fermentation process has been proposed to improve the organoleptic characteristics of wines. In the present study the fermentation performances and the interactions of mixed and sequential cultures of Hanseniaspora uvarum, Candida zemplinina, and a strain of Saccharomyces cerevisiae isolated from organic musts were investigated. To evaluate the oenological performances of the tested strains microvinifications in pasteurized red grape juice from Montepulciano d’Abruzzo cultivar were compared. The course of fermentation has been controlled through classical determinations (CO2 evolution, ethanol, glycerol, pH, total titratable acidity, sugar content, free sulfur dioxide (SO2), dry extract, sugars, organic acids, and volatile compounds). Moreover, the yeast population was determined by both culture-dependent and independent approaches. In particular, the pure culture of H. uvarum and C. zemplinina did not end the fermentation. On the contrary, when S. cerevisiae was added, fermentations were faster confirming that yeast interactions influence the fermentation kinetics. Moreover, C. zemplinina showed a good interaction with S. cerevisiae by increasing the fermentation kinetic in high gravity Montepulciano must, with low ethyl acetate and acetic acid production. This study confirmed that non-Saccharomyces yeasts play a crucial role also in organic wines and their activity could be modulated through the selection of appropriate strains that correctly interact with S. cerevisiae