Tracing the identity of Parmigiano Reggiano "Prodotto di Montagna - Progetto Territorio" cheese using NMR spectroscopy and multivariate data analysis.

BACKGROUND: Nuclear magnetic resonance (NMR) spectroscopy is one of the well-established tools for food metabolomic analysis, as it proved to be very effective in authenticity and quality control of dairy products, as well as to follow product evolution during processing and storage. The analytical assessment of the EU mountain denomination label, specifically for Parmigiano Reggiano "Prodotto di Montagna - Progetto Territorio" (Mountain-CQ) cheese, has received limited attention. Although it was established in 2012 the EU mountain denomination label has not been much studied from an analytical point of view. Nonetheless, tracing a specific profile for the mountain products is essential to support the value chain of this specialty. RESULTS: The aim of the study was to produce an identity profile for Parmigiano Reggiano "Prodotto di Montagna - Progetto Territorio" (Mountain-CQ) cheese, and to differentiate it from Parmigiano Reggiano PDO samples (conventional-PDO) using 1H NMR spectroscopy coupled with multivariate data analysis. Three different approaches were applied and compared. First, the spectra-as-such were analysed after proper preprocessing. For the other two approaches, Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) was used for signals resolution and features extraction, either individually on manually-defined spectral intervals or by reapplying MCR-ALS on the whole spectra with selectivity constraints using the reconstructed "pure profiles" as initial estimates and targets. All approaches provided comparable information regarding the samples' distribution, as in all three cases the separation between the two product categories conventional-PDO and Mountain-CQ could be highlighted. Moreover, a novel MATLAB toolbox for features extraction via MCR-ALS was developed and used in synergy with the Chenomx library, allowing for a putative identification of the selected features. SIGNIFICANCE: A first identity profile for Parmigiano Reggiano "Prodotto di Montagna - Progetto Territorio" obtained by interpreting the metabolites signals in NMR spectroscopy was obtained. Our workflow and toolbox for generating the features dataset allows a more straightforward interpretation of the results, to overcome the limitations due to dimensionality and to peaks overlapping, but also to include the signals assignment and matching since the early stages of the data processing and analysis.

Behaviour of Listeria monocytogenes in packaged water buffalo mozzarella cheese.

AIMS: The aim of the study was to evaluate the behaviour of Listeria monocytogenes in the conditioning liquid of packaged water buffalo mozzarella cheese (WBMC). METHODS AND RESULTS: The conditioning liquid was contaminated with L. monocytogenes, and the contaminated samples were stored at four different storage temperatures: 5 and 10 °C for 22 days; 20 °C for 9 days; 20 °C for 3 days and then at 5 °C for 6 days. The results showed that L. monocytogenes concentration decreased when contaminated samples were stored at 5 °C. When WBMC was stored at 20 °C and at 10 °C, L. monocytogenes started to grow after a lag phase of 3 and 10 days, respectively. When samples were stored at variable temperature conditions, L. monocytogenes numbers showed a lag phase of 5 days. CONCLUSIONS: Use of a conditioning liquid characterized by acidity and a correct storage temperature is able to counteract pathogen replication during shelf life. A high concentration of lactic acid bacteria was associated with effective control of L. monocytogenes but the role of lactic acid bacteria in WBMC conditioning liquid requires further investigation. SIGNIFICANCE AND IMPACT OF THE STUDY: According to European regulations, food producers should be able to justify decision-making on the shelf life assigned to their products, taking into account reasonable storage conditions and use by consumers. The results of the trial yielded information for producers of WBMC and similar cheeses for decision-making on product shelf life.

Isolation of glucosidase and phospholipase positive Bacillus circulans on ALOA medium.

AIM: To report the growth of glucosidase and phospholipase positive bacteria on agar Listeria according to Ottaviani and Agosti (ALOA) different from Listeria monocytogenes, Listeria ivanovii and Bacillus cereus. METHODS AND RESULTS: Raw water-buffalo milk was analysed according to EN ISO 11290. Streaking of Fraser broth on ALOA resulted in green colonies with an opaque halo after 48 h at 30°C. Colonies were transferred onto Tryptone soya yeast extract agar and showed cultural characteristics atypical for L. monocytogenes. Results of confirmation tests according to EN ISO 11290 method: negative haemolysis test, weak positive camp test in correspondence with Staphylococcus aureus, no fermentation of rhamnose, fermentation of xylose. Gram staining showed tapered, curved, Gram-positive rods with subterminal to terminal ellipsoidal spores, 0.5-0.7 µm diameter 4-5 µm. API 50CH CHB kit (99.9% percentage of identification) and the sequence of the 833 bp PCR product (portion of 16S rRNA, generic primers 1492-r; p27-f) showed 97% identity with Bacillus circulans ATCC 4513 (GenBank AY724690). CONCLUSIONS: Some B. circulans strains can grow on ALOA, according to ISO 11290, confirmation tests readily differentiate B. circulans from L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The different morphology of the colonies must be kept in mind to select true L. monocytogenes for confirmation test and to avoid overestimation of L. monocytogenes count.

Intestinal metabolism of weaned piglets fed a typical United States or European diet with or without supplementation of tributyrin and lactitol.

The aim of the study was to investigate the effects of supplementation of a microencapsulated blend of tributyrin and lactitol (TL) to a standard European (EU) diet without antibiotic growth promoters on intestinal metabolism and mucosa development of weaned piglets and to compare it with a standard US diet containing animal proteins, zinc oxide, copper sulfate, and carbadox. Ninety piglets weaned at 21 d were divided into 3 dietary groups consisting of 5 replicates each: 1) US diet supplemented with 55 mg/kg of carbadox, and 2.5% each of plasma proteins and spray-dried blood cells in the first phase, 3,055 mg/kg of Zn in the first and second phases, and 180 mg/kg of Cu in the third phase; 2) EU diet based on vegetable proteins and no antibiotics; and 3) the same EU diet supplemented with 3,000 mg/kg of microencapsulated TL. The study was divided into 3 phases: 0 to 7, 8 to 21, and 22 to 35 d. On d 7, 21, and 35, animals were weighed, and feed consumption and efficiency were determined. On d 14 and 35, one pig per pen was killed, and the intestinal contents and mucosa from the proximal, middle, distal jejunum and the ileum were sampled. Intestinal wall sections were fixed for histological analysis, and intestinal content was used for VFA, ammonia, and polyamine analysis. Throughout the study (d 0 to 35), the US diet had greater ADG and ADFI than the EU diet (P < 0.05). The EU diet supplemented with TL tended to have 11% greater ADG (P = 0.17). Feeding the EU diet caused a reduction in proximal and middle jejunum villi length by 10% (P < 0.05) and an increase in crypt size in proximal jejunum (P < 0.05) compared with the US diet, probably due to an increased rate of cell loss and crypt cell production. The TL supplementation resulted in longer villi along the jejunum and less deep crypts in the proximal jejunum (+15.9 and -8.9%, respectively; P < 0.05) than the unsupplemented EU diet. The TL diet increased the concentrations of cadaverine and putrescine in the small intestine (P < 0.05) and seemed to increase cadaverine, histamine, putrescine, and spermine in the large intestine by 1.5- to 10-fold compared with the US or EU diet. In conclusion, although the US diet had a greater effect on growth performance and mucosal trophic status than the EU diets, the supplementation with slowly released TL seemed to be an effective tool to partially overcome the adverse effects of vegetable protein diets.

Lipid microencapsulation allows slow release of organic acids and natural identical flavors along the swine intestine.

The purpose of the present work was to investigate the in vivo concentrations of sorbic acid and vanillin as markers of the fate of organic acids (OA) and natural identical flavors (NIF) from a microencapsulated mixture and from the same mixture non-microencapsulated, and the possible consequences on the intestinal microbial fermentation. Fifteen weaned pigs were selected from 3 dietary groups and were slaughtered at 29.5 +/- 0.27 kg of BW. Diets were (1) control; (2) control supplemented with a blend of OA and NIF microencapsulated with hydrogenated vegetable lipids (protected blend, PB); and (3) control supplemented with the same blend of OA and NIF mixed with the same protective matrix in powdered form but without the active ingredient coating (non-protected blend, NPB). Stomach, cranial jejunum, caudal jejunum, ileum, cecum, and colon were sampled to determine the concentrations of sorbic acid and vanillin contained in the blend and used as tracers. Sorbic acid and vanillin were not detectable in pigs fed the control, and their concentrations were not different in the stomach of PB and NPB treatments. Pigs fed PB showed a gradual decrease of the tracer concentrations along the intestinal tract, whereas pigs fed NPB showed a decline of tracer concentration in the cranial jejunum and onwards, compared with the stomach concentrations. Sorbic acid and vanillin concentrations along the intestinal tract were greater (P = 0.02) in pigs fed PB compared with pigs fed NPB. Pigs fed PB had lower (P = 0.03) coliforms in the caudal jejunum and the cecum than pigs fed the control or NPB. Pigs fed the control or PB had a greater (P = 0.03) lactic acid bacteria plate count in the cecum than pigs fed NPB, which showed a reduction (P = 0.02) of lactic acid concentrations and greater (P = 0.02) pH values in the caudal jejunum. The protective lipid matrix used for microencapsulation of the OA and NIF blend allowed slow-release of both active ingredients and prevented the immediate disappearance of such compounds upon exiting the stomach.