The Effects of Encapsulation on the In Vitro Anti-Clostridial Activity of Olive Mill Wastewater Polyphenolic Extracts: A Promising Strategy to Limit Microbial Growth in Food Systems.

Despite the technologies applied to food production, microbial contamination and chemical deterioration are still matters of great concern. In order to limit these phenomena, new natural approaches should be applied. In this context, the present study aimed to assess the antioxidant and anti-Clostridial effects of two different polyphenolic extracts derived from olive mill vegetation water, one liquid (LE) and one encapsulated (EE). The extracts have been preliminary characterized using Liquid Chromatography Quadrupole Time-Of Flight spectrometry. The Oxygen Radical Absorbance Capacity method was used to determine the antioxidant capacity, registering a higher value for EE compared to that for LE (3256 ± 85 and 2446 ± 13 µgTE/g, respectively). The antibacterial activity against C. perfringens, C. botulinum and C. difficile was studied by the agar well diffusion method, MIC and MBC determination and a time-kill test. The results confirm that EE and LE are able to limit microbial growth, albeit with minor effects when the phenolic compounds are encapsulated. Further studies are needed to evaluate the possible application of these extracts in food systems.

Occurrence of Hydroxytyrosol, Tyrosol and Their Metabolites in Italian Cheese.

Tyrosol (T) and hydroxytyrosol (HT) are phenyl alcohol polyphenols with well-recognized health-promoting properties. They are widely diffused in several vegetables, especially in olive products (leaves, fruits and oil). Therefore, they could be present in food produced from herbivorous animals such as in milk and cheese. In this study, an analytical method to determine T, HT and some of their phase II metabolites (sulphates and glucuronides) in cheese was developed and validated. Samples were extracted with an acidic mixture of MeOH/water 80/20 (v/v) and, after a low temperature clean-up, the extracts were evaporated and injected in a liquid-chromatography coupled with high resolution mass spectrometry (LC-Q-Orbitrap). A validation study demonstrated satisfactory method performance characteristics (selectivity, linearity, precision, recovery factors, detection and quantification limits). The developed protocol was then applied to analyze 36 Italian cheeses made from ewe, goat and cow milk. The sum of detected compounds (T, tyrosol sulfate, hydroxytyrosol-3-O-sulfate and hydroxytyrosol-4-O-sulfate) reached as high as 2300 µg kg-1 on a dry weight basis, although in about 45% of cow cheeses it did not exceed 50 µg kg-1. Ewe cheeses were significantly richer of polyphenols (sum) as well as HT sulfate metabolites than cow cheeses. In conclusion, results shows that cheese cannot be considered an important dietary source of these valuable compounds.

Estimation of acrylamide exposure in Italian schoolchildren consuming a canteen menu: health concern in three age groups.

Acrylamide (AA) is a food process contaminant with carcinogenic and genotoxic properties that is formed in thermally treated foods, especially carbohydrate-rich. Dietary exposure to AA, due to school canteen foods, was estimated in schoolchildren aged 3-13 years by combining the AA concentration in foods with the amount and frequency of food consumption. Potato products and bakery products presented the highest mean levels of AA (841 and 244 µg/kg, respectively) followed by meat (222 µg/kg) and egg products (151 µg/kg). The mean total AA intake was estimated at 2.16 µg/kg bw/d, with the highest percentages provided by potato products (34.5%), meat products (26.1%) and bread (24.5%). The calculated margins of exposure (MOEs) for the average AA exposure (84 and 212 for the benchmark dose lower confidence limits (BMDL) of 0.17 and 0.43 mg/kg bw/d) suggest that there is a health concern with respect to students eating in school canteens.

Comparison of PCR assays targeting the multi-copy targets B1 gene and 529 bp repetitive element for detection of Toxoplasma gondii in swine muscle.

The comparison of the sensitivities of two molecular assays designed to target the multi-copy sequences of the Toxoplasma gondii genomic B1 region and 529 bp-RE respectively, in detecting T. gondii in swine muscle was assessed. Diaphragm pillars were obtained from 498 slaughtered pigs managed in intensive farms in Central Italy. Genomic DNA was extracted from the tissues and T. gondii-B1 and 529 bp-RE sequences were amplified by specific PCR protocols. Toxoplasma gondii DNA was detected in 165 samples (33.13%). There was a good correlation (κ = 0.77) between the results obtained targeting the two different genetic markers, however the 529 bp RE-PCR assay overall detected a significantly higher (P < 0.05) number of T. gondii-positive samples (150 samples) than the B1-PCR protocol (134). Our results show that: i) standardized B1 and 529 bp-RE PCRs applied to muscle tissues can detect a high rate of T. gondii-infection; ii) a multi-target PCR approach is recommended for the accurate diagnosis of infection in swine and can also be used in food testing.

Seasonal variations of Italian Mediterranean Buffalo (Bubalus bubalis) Mozzarella cheese quality.

Water buffalo Mozzarella cheese is more appreciated in the summer, but milk production is higher during wintertime, as water buffalo are seasonally polyoestrous animals. The aim of the study reported in this Research Communication was to evaluate the effect of the reversing of the calving period on Mozzarella cheese composition and quality traits. Ten batches of Mozzarella cheeses produced during the summer and winter periods were analysed for pH, colour, chemical composition. Seasonal differences were observed for cheese yield (26·66% in winter and 25·61% in summer), moisture content (66·54% in winter and 61·18% in summer) colour and consumer evaluation.

Influence of Maternal and Postweaning Linseed Dietary Supplementation on Growth Rate, Lipid Profile, and Meat Quality Traits of Light Sarda Lambs.

The effects of dietary extruded linseed (EL) on growth performance, meat quality, and lipid profile of Semimembranosus and Longissimus lumborum muscles of 81 Sarda lambs were studied in a 3 × 3 design: EL content (0%, 10%, and 20%) of maternal dietary concentrate fed from 20 d to parturition to 60 d of lactation and EL content (0%, 10%, 20%) of lamb concentrate fed after weaning for 30 d. The basal diet was composed of alfalfa and meadow hay during pregnancy and alfalfa hay during lactation. At slaughter, carcass and meat quality were evaluated. Sensory quality of Semimembranosus from 0% and 20% EL lambs was assessed. Both maternal and postweaning diets affected growth performance, with higher body weights recorded with the 10% EL concentrate. Carcass and meat quality were not affected by diet. Saturated and monounsaturated FA decreased and n-3 polyunsaturated FA increased with increasing EL content in lamb diet. An increase in vaccenic and rumenic acid was associated with the EL content of the maternal diet. Both diets increased the n-6/n-3 FA ratio. No differences in acceptability were detected by consumers among groups. It is concluded that EL supplementation and early life nutrition can influence performance and FA metabolism in growing lambs.

Influence of manufacturing procedure on the compositional and sensory properties of n-3 fatty acid-enriched pecorino cheese.

Pecorino cheeses were produced using two different manufacturing procedures (either from raw milk or from milk previously subject to a thermisation at 65 °C for 10 s with the addition of starter culture) using the milk obtained from three groups of ewes fed three different concentrates: (1) control, (2) control enriched with 100 g/kg of extruded linseed, and (3) control enriched with 200 g/kg of extruded linseed. All concentrates were administered with alfalfa hay ad libitum. The extruded linseed-enriched diets increased the concentration of n-3 fatty acids, conjugated linoleic acid, and monounsaturated and polyunsaturated acids in raw milk and thermised milk cheese and decreased the concentration of saturated fatty acids. Changes in the manufacturing procedure did not influence the fatty acid composition of the cheeses but markedly influenced their sensory properties. In particular, heat-treating the milk and adding a starter reduced the differences in cheese odour, flavour and toughness induced by the diet when raw milk was used. Cheeses made with thermised milk and the addition of starter culture were more uniform and obtained a higher score in the preference test compared with the corresponding raw milk cheeses. An appropriate manufacturing process, therefore, can be a possible strategy to obtain cheeses with improved health-promoting properties and an unaltered acceptance level by consumers.

Preventing Microbial Growth in Game Meat by Applying Polyphenolic Extracts from Olive Mill Vegetation Water.

We studied the efficacy of different formulations of polyphenol extracts (mainly containing hydroxytyrosol and tyrosol) from olive mill vegetation water on the microflora on the surfaces of game meat cuts with high or low initial bacterial loads. Meat with a high microbial load (>5 Log cfu/g; mean value = 6.83 ± 0.45 standard deviation) was immersed for 10 or 60 sec into 25% and 10% solutions of microencapsulated freeze-dried and non-encapsulated polyphenolic extracts. Aerobic colony, Enterobacteriaceae, Pseudomonas spp., and lactic acid bacteria counts were determined on treated samples compared to controls after 7 days of storage (in vacuum-packed conditions at +3 °C). Significant differences were registered only for aerobic colony count for a 10% liquid extract treatment (0.64 log reduction). In contrast, the dipping or immersion of game meat with low initial microbial loads (<5 Log cfu/g; mean value = 3.58 ± 0.72 standard deviation) in 10% solutions of the polyphenol extracts effectuated significant reductions in all bacteria counts (p < 0.002) at 7 and 14 days of storage for different extracts, independently from the application methods. The use of the extracts to inhibit bacterial growth in game meat should only be considered if a good hygienic baseline is guaranteed.

Surface carcass treatment with olive mill wastewater polyphenolic extract against Salmonella Enteritidis and Listeria monocytogenes: in vitro and in situ assessment.

In recent years, there has been an increased interest in substances that could inhibit or reduce microbial growth in food products. Olive oil industry by-products, due to bioactive compounds with potential antimicrobial properties such as polyphenols, could be used in carcass treatment to enhance hygienic and quality traits. The assessment of the antimicrobial efficacy of bioactive molecules against pathogens should be determined with in vitro and in situ models since it is not possible to evaluate it directly on carcasses at the slaughterhouse. This study aimed to evaluate the effect of an olive mill wastewater polyphenolic extract against Salmonella Enteritidis and Listeria monocytogenes, simulating carcass surfaces using bovine dermis samples that were experimentally contaminated with the selected pathogens. The minimum inhibitory concentration and minimum bactericidal concentration were first determined for S. Enteritidis and L. monocytogenes. In situ, bactericidal activity assessment was performed using 20 cm2 derma samples contaminated with 5 Log CFU/20 cm2 of S. Enteritidis and L. monocytogenes in separate trials. Treatment with the polyphenolic extract was not effective for either microorganism. In order to establish the bacteriostatic activity of the polyphenolic extract, suspensions of about 2 Log CFU/20 cm2 of S. Enteritidis and L. monocytogenes were used. Polyphenolic extract treatment was not effective against Salmonella, while for Listeria it allowed microbial growth to delay (around 1 Log CFU/cm2 difference at 3, 7, and 14 days between treated and control groups). Further investigations are needed to evaluate the application of polyphenolic compounds on carcass surfaces and their effects on sensory traits.

Antilisterial activity of olive-derived polyphenols: an experimental study on meat preparations.

Pork meat and processed pork products have been linked to multiple listeriosis outbreaks worldwide during the past few years. Specifically, it has been highlighted that minced pork meat is easily perishable and may increase the growth of Listeria monocytogenes, which could be harmful to the general public's health. This study aimed to investigate the potential application of olive oil mill wastewater polyphenolic and red beet extracts as natural antimicrobial agents for L. monocytogenes growth control in burgers. The minced pork meat was mixed with the extracts and experimentally inoculated with L. monocytogenes, then molded into vacuum-packaged and cold-stored (4±1°C) burgers kept under alternating exposure to fluorescent light. The L. monocytogenes enumeration was performed on burgers at 0, 2, 5, and 10 days of shelf life. In uninoculated burgers, physicochemical (pH, water activity, color) and sensory determination (descriptive sensory analysis) were also conducted. At the end of storage, the samples treated with olive-derived extract showed the lowest value of L. monocytogenes (approximately 1.3 Log CFU/g). The physicochemical and sensory traits of burgers have benefited from the addition of both olive-derived and red beet extracts. Results suggest that olive mill wastewater polyphenolic extracts could be added to minced pork meat products to act as a natural antimicrobial agent.