The Occurrence of Beer Spoilage Lactic Acid Bacteria in Craft Beer Production

Beer is one of the world's most ancient and widely consumed fermented alcoholic beverages produced with water, malted cereal grains (generally barley and wheat), hops, and yeast. Beer is considered an unfavorable substrate of growth for many microorganisms, however, there are a limited number of bacteria and yeasts, which are capable of growth and may spoil beer especially if it is not pasteurized or sterile‐filtered as craft beer. The aim of this research study was to track beer spoilage lactic acid bacteria (LAB) inside a brewery and during the craft beer production process. To that end, indoor air and work surface samples, collected in the brewery under study, together with commercial active dry yeasts, exhausted yeasts, yeast pellet (obtained after mature beer centrifugation), and spoiled beers were analyzed through culture‐dependent methods and PCR‐DGGE in order to identify the contaminant LAB species and the source of contamination. Lactobacillus brevis was detected in a spoiled beer and in a commercial active dry yeast. Other LAB species and bacteria ascribed to Staphylococcus sp., Enterobaceriaceae, and Acetobacter sp. were found in the brewery. In conclusion, the PCR‐DGGE technique coupled with the culture‐dependent method was found to be a useful tool for identifying the beer spoilage bacteria and the source of contamination. The analyses carried out on raw materials, by‐products, final products, and the brewery were useful for implementing a sanitization plan to be adopted in the production plant.     

Digestive protection of probiotic Lacticaseibacillus rhamnosus in Ricotta cheese by monoglyceride structured emulsions

This research aimed at studying the potential use of monoglyceride (MG) structured emulsions (MSEs) as delivery and protective systems for probiotic bacteria in Ricotta cheese. To this purpose, a low-fat commercial Ricotta cheese was added with MSEs formulated with milk, as water phase, and sunflower oil (MSE-SO) or anhydrous milk fat (MSE-AMF), as lipid phase. A commercial whole milk Ricotta cheese (W-RC) was considered as reference. A probiotic Lacticaseibacillus rhamnosus strain was inoculated as free cells in W-RC or embedded into the MSEs and added to the low-fat Ricotta at the same reference fat content. After physico-chemical characterisation, L. rhamnosus viability and sample destructuring behaviour upon in vitro digestion were evaluated. At the end of in vitro digestion, both W-RC and sample containing MSE-SO were unable to protect cells. By contrast, sample with AMF ensured a sufficient probiotic viability, even after 14 days of storage at 4 °C. This result was attributed to system composition and structure. During the gastric phase, the presence of caseins and MG-AMF mixed structures induced the formation of clots, entrapping and protecting cells against the acidic pH of the stomach, as confirmed by confocal micrographs and particle size. During the intestinal phase, cell viability was guaranteed by the formation of mixed micelles promoted by MG. It was demonstrated that microbial cells located near MG structures where they found protection.     

Application of the novel Droplet digital PCR technology for identification of meat species

The authenticity and traceability of meat products are issues of primary importance to ensure food safety. Unfortunately, food adulteration (e.g. the addition of inexpensive cuts to minced meat products) and mislabelling (e.g. the inclusion of meat from species other than those declared) happens frequently worldwide. The aim of this study was to apply a droplet digital PCR assay for the detection and quantification (copies μL⁻¹) of the beef, pork, horse, sheep, chicken and turkey in meat products. The analysis conducted on commercial meat showed the presence of traces of DNA from other animal species than those declared. We show that the method is highly sensitive, specific and accurate (accuracy = 100%). This method could be adopted by competent food safety authorities to verify compliance with the labelling of meat products and to ensure quality and safety throughout the meat supply chain, from primary production to consumption.     

Characterisation of Staphylococcus aureus strains from milk and goat cheese and evaluation of their inhibition by gallic acid,nisin and velame of the Brazilian caatinga

Twenty isolates from milk and goat cheese were confirmed as Staphylococcus aureus. These isolates were characterised for phenotypic properties related to cell adhesion and for the presence of enterotoxin production, intercellular adhesion and β‐lactam resistance genes. Staphylococcus aureus L47 showed cell adhesion ability and positivity for the sec, sed, icaD, mecA and blaZ genes. Three antimicrobial compounds were tested singly or in pairs for growth control of strain L47: gallic acid (GA), nisin and essential oil (EO) of Croton heliotropiifolius (velame). At 24 h, EO and EO + nisin showed higher inhibitory activity against S. aureus L47 in goat milk.     

External Shape Differences between Sympatric Populations of Commercial Clams <Emphasis Type="Italic">Tapes decussatus</Emphasis> and <Emphasis Type="Italic">T. philippinarum</Emphasis>

The actual Italian production of clams is chiefly sustained by the native Tapes decussatus and the fortuitously imported Tapes philippinarum. Both species are commercialized as “Vongola verace”, but the commercial value of T. philippinarum is lower. The discrimination of species by sight is usually difficult and it cannot be done by observation based on shell morphology but only when animals open their valves hence displaying the two siphons. In this study, we propose a new, noninvasive method to discriminate individuals of both species based on the analysis of the external shape of their shells. Accordingly, in sympatric populations at two sites of the Po river outlet, we have chosen individuals (63 for T. decussatus and 57 for T. philippinarum) of comparable commercial size for which a certain genetic discrimination was previously done. Pictures of the left side valve were taken for all specimens. Their profiles were analyzed with the elliptic Fourier analysis. The mean outline for each species was graphically extracted. The coefficients of the harmonic equations were analyzed by multivariate classification (partial least squares discriminant analysis [PLSDA]). Results showed a high percentage of correct classification of individuals of both species (96.6%). Contour analysis reflected the overall shell shape and thus identified morphological aspects that were difficult to recognize and quantify in sight. The high percentage of correct classifications obtained by combining the analysis of elliptic Fourier harmonics with PLSDA demonstrated the feasibility of this method to discriminate species with a high level of resemblance.     

Isolation and molecular characterization of antibiotic-resistant lactic acid bacteria from poultry and swine meat products

The transfer via the food chain from animals to humans of microbes that are resistant to antimicrobial agents is of increasing concern. To determine the contributions of nonpathogenic microflora to the occurrence and spread of antibiotic resistance (AR) genes in the food chain, 123 lactic acid bacteria were isolated from 29 samples of raw and processed pork and chicken meat products that had previously tested positive for one or more AR genes that encode clinically relevant ARs: tet(M), tet(O), tet(K), erm(A), erm(B), erm(C), aac (6')-Ie aph (2")-Ia, mecA, and blaZ. All of the isolates were initially tested for their AR gene profiles by PCR. The 59 isolates carrying a tet, erm, or blaZ gene were taken through molecular identification, analyzed by determination of the MIC, and subjected to genetic fingerprinting. Lactococcus garvieae was the predominant species (28 isolates), followed by Lactobacillus plantarum (11 isolates) and L. salivarius (6 isolates), whereas Lactococcus lactis subsp. lactis, Lactobacillus johnsonii, L. reuteri, L. crispatus, and L. brevis were identified at lower frequencies. The tet(M) and erm(B) genes were the most frequently detected. Assessment of multiple resistances in 18 tet positive (tet+) isolates revealed that tet(M) plus erm(B) and tet(K) plus erm(B) were the most frequent AR gene patterns. Partial sequencing of the tet(M) open reading frame of three selected strains showed high sequence similarities (> 99%) with tet(M) genes previously found in human pathogens (Listeria monocytogenes and Neisseria meningitidis). Southern hybridization with plasmid profiles revealed these strains contained tet(M)-carrying plasmids.     

Role of surface-inoculated Debaryomyces hansenii and Yarrowia lipolytica strains in dried fermented sausage manufacture. Part 1: Evaluation of their effects on microbial evolution, lipolytic and proteolytic patterns

The aim of this work was to study the effects of Debaryomyces hansenii and Yarrowia lipolytica strains, used with lactic acid starter cultures (Lactobacillus plantarum), on the manufacture of dried fermented sausages to understand their role on sausage microbial evolution, lipolytic and proteolytic patterns. The inoculation of the yeast strains did not markedly affect the sausage’s microbial flora. The sausages with the yeast strains showed more marked and earlier water activity (a_w) reductions. Moreover, the surface inoculation of the yeast strains resulted, at the end of ripening, in more pronounced proteolysis and lipolysis. The lipolytic patterns of the products were affected not only by the yeast strain but also by the level of mincing of the meat mixture used.     

Proton binding onto soil by nonelectrostatic models: isolation and identification of mineral contributions

In this paper a methodological approach is proposed to validate mechanistic modeling for proton binding onto active sites of mineral and soil samples by reducing the uncertainty and arbitrariness of model schematization. This approach is based on the quantitative formulation (X-ray calibration method) of a simulating mineral mixture (SMM) accounting for the main mineral phases in the soil (quartz, goethite, hematite, muscovite, clinochlore). Mineral and organic contributions were separated by comparing titration curves of river sediment and SMM. Specific mineral contributions to the acid properties of SMM were separated by comparing titration models of SMM and single minerals. Different nonelectrostatic models were used for titrations of SMM and single minerals: two-site/three-KH models (one amphoteric plus one monoprotic site) for clay minerals and SMM; one-site/two-KH models (one amphoteric site) for goethite and hematite; and a one-site/one-KH model (one monoprotic site) for quartz. Crossed-comparisons of titration models allow for identifying and quantifying the specific contributions of the distinct edge hydroxyl groups of iron oxides, clay minerals, and quartz in the different pH ranges. In particularthe amphoteric sites of aluminosilicates mainly contribute in the acid-neutral pH range, the amphoteric sites of iron oxides take part in the neutral-basic range, and finally the monoprotic edge hydroxyl groups of quartz react in the upper basic region of pH. The good simulation of the acid-base properties of SMM (according to single mineral titration models and quantitative composition by X-ray) confirms both model schematization and SMM formulation. Speciation diagrams of the active sites of the different mineral components (aluminosilicates, iron oxides, and quartz) were obtained by implementing the database of a dedicated software with the apparent equilibrium constants regressed by titration modeling of single minerals.     

Effect of dry‐heat treatments on the nutritional value of maize germ

Maize germ is a by‐product of the maize milling process that is characterised by a high nutritional value. Currently, heat treatments are employed to prevent full‐fat maize germ from spoilage. The aim of this research was to study the effect of five dry‐heat treatments on the nutritional value of full‐fat maize germ. The results confirmed that after each dry‐heat treatment, the lipase activity decreases but the use of high temperatures could be detrimental for phytosterol and thiamine concentrations. The main negative effects have been observed after treatments at 140 °C for 30 min and 160 °C for 10 min. No significant difference has been observed for protein, ash or fatty acid contents. The treatment at 140 °C for 20 min resulted an optimal combination between temperature and heating time to inactivate lipase without altering deeply the nutritional value and the colour of maize germ.