Reduction of PDO Pecorino Siciliano cheese making duration: Microbial dynamics and quality attributes deriving from replacing whey permeate with hot water during cooking.

This work was carried out with the aim to reduce the transformation duration of Protected Designation of Origin (PDO) Pecorino Siciliano cheese. To this purpose, the cooking in hot water (experimental production, EXP) was compared to the traditional cheese cooking under whey permeate (control production, CTR). The microbiological composition of under rind (UR) and core (Co) section of CTR and EXP cheeses was determined by a combined culture-dependent and -independent approach. Total mesophilic microorganisms and lactic acid bacteria (LAB) present in raw ewes' milk (5.0 log CFU/mL) increased during cheese making and reached values of about 8.0 log CFU/g in both sections (UR and Co) of 5-month ripened cheeses of both productions (CTR and EXP) monitored. The identification of the viable LAB populations in ripened cheeses showed that Enterococcus, Lacticaseibacillus, Lactiplantibacillus, Levilactobacillus, Limosilactobacillus and Streptococcus dominated UR and Co sections of all cheeses. MiSeq Illumina analysis demonstrated that LAB populations (lactobacilli, lactococci and streptococci) dominated the bacterial community of cheeses at 95.63-98.41 % of relative abundance. The two different cooking operations did not influence the physicochemical characteristics of PDO Pecorino Siciliano cheeses. Sensory evaluation performed by artificial senses analysis and trained panelists confirmed that the modification of PDO Pecorino Siciliano cheese production protocol did not significantly affect product characteristics and overall acceptance. Thus, data of this work confirmed that cooking under hot water allowed to reduce transformation duration and safeguard typicality of PDO Pecorino Siciliano cheese.

Effect of commission implementing regulation (EU) 2020/1319 on the bacterial composition of PDO Provola dei Nebrodi cheese.

In this study, PDO Provola dei Nebrodi cheese was deeply characterized for its bacterial community and chemical composition. Four dairy factories (A-D) were monitored from milk to ripened cheese. Wooden vat biofilms were dominated by thermophilic rod LAB (4.6-6.5 log CFU/cm2). Bulk milk showed consistent levels of total mesophilic microorganisms (TMM) (5.0-6.0 log CFU/mL) and, after curdling, a general increase was recorded. The identification of the dominant LAB in wooden vat biofilms and ripened cheeses showed that the majority of wooden vat LAB were lactococci and Streptococcus thermophilus, while cheese LAB mainly belonged to Lacticaseibacillus paracasei and Enterococcus. Illumina sequencing identified 22 taxonomic groups; streptococci, lactococci, lactobacilli and other LAB constituted the majority of the total relative abundance % of the wooden vat (69.01-97.58 %) and cheese (81.57-99.87 %) bacterial communities. Regarding chemical composition, the effect of dairy factories was significant only for protein content. Inside cheese color was lighter and yellower than surface. Differences in fatty acids regarded only myristic acid and total amount of monounsaturated fatty acids. The sensory evaluation indicated some differences among cheeses produced in the four dairies regarding color, homogeneity of structure, overall intensity, salty, spicy, and hardness. The integrated approach applied in this study showed that PDO Provola dei Nebrodi cheese characteristics are quite stable among the dairy factories analyzed and this has to be unavoidably imputed to the application of the same cheese making protocol among different dairies.

Chemometric Approaches to Analyse the Composition of a Ewe's Colostrum.

Colostrum is a major source of immunity in lambs and, in general, in all newborn ruminants. It allows the transfer of antibodies from the ewe to the lamb, and it becomes the exclusive source of nutrients for a newborn. Among the most significant Pearson correlations, the positive correlation between the Brix refractive index (Brix) and protein (0.90) should be noted. Both parameters (protein percentage and Brix) were then positively correlated with the percentage of fat (0.38 and 0.41), urea (0.81 and 0.67), calcium (0.39 and 0.29), and magnesium (0.58 and 0.59), as well as the yellowness (0.78 and 0.75). Somatic cell count (SCC) and pH, parameters which are indicators of subclinical mastitis, were positively correlated (0.49), and SCC was positively correlated with sodium (0.37) and negatively correlated with potassium (-0.28). Among the macroelements in colostrum-calcium, potassium, magnesium, and sodium-the correlations were largely positive. With respect to the colour parameters, yellowness was negatively correlated with lightness (-0.41) and redness (-0.45). The factor analysis split the total of variance into three latent factors. The first factor was named "Colostrum quality of grazing sheep" because it was positively correlated primarily with SCC, pH, Poly Unsatured Fatty Acids (PUFA), and the sum of the omega-3 Fatty Acids (FAs). The second factor, named "Good quality colostrum", was positively correlated primarily with the refractive index, protein and fat percentages, urea content, phosphorus, Mono Unsatured Fatty Acids (MUFA), and yellowness. The third factor was positively correlated primarily with calcium, potassium, magnesium, and sodium, and it was therefore termed "Mineral component of colostrum". Stepwise discriminant analysis showed that the protein percentage, calcium, and magnesium were able to explain more than 85% of the Brix refractive index, which remains a good parameter for simply evaluating the nutritional quality of sheep colostrum at the level of a farm.

Feeding Dairy Ewes with Fresh or Dehydrated Sulla (Sulla coronarium L.) Forage. 1. Effects on Feed Utilization, Milk Production, and Oxidative Status.

Feeding dairy ewes with fresh sulla forage (FSF), a legume species containing condensed tannins (CT), has been shown to increase feed intake, milk yield, and casein and enhances the oxidative status of animals. Dehydration of FSF could be an alternative to hay-making to preserve the nutritional properties. This research aimed to compare the responses of dairy ewes fed with diets based on sulla hay (SH), pelleted dehydrated sulla forage (DSF), or FSF in terms of efficiency of feed utilization, milk production, and the balance between oxidant (reactive oxygen metabolites (ROMs)) and antioxidant (biological antioxidant potential (BAP)) substances at the plasma level. Ten first-lambing (FL) and ten third-lambing (TL) ewes of the Valle del Belice breed at 60 days in milk were allocated into five homogeneous groups fed with different diets in a partial 5 × 2 Latin square design with two phases. The diets differed for the forage basis: SHL = SH ad libitum; DSF2 = 2 kg/day DSF per head plus SH ad libitum; FSF2 = 2 kg/day FSF per head plus SH ad libitum; FSF4 = 4 kg/day FSF per head plus SH ad libitum; FSFL = FSF ad libitum. A commercial concentrate was provided to FL (0.8 kg/day per head) and TL (1.2 kg/day per head) ewes. Dehydration induced slight variations in the content of protein and fiber, showed no loss of vitamin E and polyunsaturated fatty acids, and decreased the level of CT and polyphenols. The DSF2 diet resulted in a higher intake of dry matter, protein, and vitamin E compared to the other diets, whereas, compared to the FSFL diet, its intake was analogous for net energy and was lower in CT and polyphenols. The DSF2 diet was comparable to FSF4 and FSFL diets for milk yield, and to all diets for casein content and the clotting ability of milk. Ewes fed a DSF2 diet exhibited lowest values of ROMs and oxidative stress index (OSI = ROMs/BAP), indicating a better oxidative status, presumably due to the antioxidant protection exerted by the higher vitamin E intake and CT metabolites. These results confirmed the positive effects of FSF on milk production, especially due to CT intake, in improving the efficiency of dietary protein utilization, and showed how favorable effects also occur when FSF is replaced by DSF instead of SH.

Feeding Dairy Ewes with Fresh or Dehydrated Sulla (Sulla coronarium L.) Forage. 2. Effects on Cheese Enrichment in Bioactive Molecules.

Sulla is a tanniferous legume species largely used as fresh or preserved forage in the ruminants' diets. Due to its content in polyphenols with antioxidant activity, especially condensed tannins (CT), fresh sulla forage (FSF), when eaten by ruminants, is able to enrich animal products with antioxidant molecules and polyunsaturated fatty acids (PUFA) that are beneficial for consumers' health. Dehydration represents a valid alternative to haymaking to preserve these properties also in periods when FSF is unavailable. In this research, the effects of ewes' diets based on sulla hay (SH), pelleted dehydrated sulla forage (DSF) or FSF were evaluated on cheese physico-chemical traits, including fatty acid (FA) profile, vitamins A and E, polyphenols, antioxidant capacity and oxidative stability. The individual daily milk from 10 first lambing (FL) and 10 third lambing (TL) Valle del Belìce ewes at about 60 days in milk, divided homogeneously into 5 groups fed different diets in a partial 5 × 2 Latin square design with 2 phases, and the bulk milk of farming ewes fed only on natural pasture, were used in 2 sessions of micro cheese-making to manufacture cheeses, sampled at 48 h of ripening. The experimental diets were: SHL = SH ad libitum; DSF2 = 2 kg/day DSF per head plus SH ad libitum; FSF2 = 2 kg/day FSF per head plus SH ad libitum; FSF4 = 4 kg/day FSF per head plus SH ad libitum; FSFL = FSF ad libitum. Concentrate was supplied at the amount of 0.8 or 1.2 kg/day per head for FL and TL, respectively. Cheese health properties greatly improved with FSFL diet, based on the exclusive use of fresh forage, that induced the increase in the content of molecules with antioxidant activity, as vitamin A, vitamin E and polyphenols, enhancing the oxidative stability, and the level of PUFA as rumenic acid (C18:2 c9t11) and α-linolenic acid (ALA, C18:3n-3). Cheeses from the DSF2 group showed levels of vitamin A, vitamin E and PUFA higher than SHL cheeses and comparable to those of FSF4 cheeses, whereas their ALA content was not different from that of FSFL cheeses. Evaluating those molecules affected by the level of fresh forage in the diet for their ability to trace the animals' feeding regime, ALA and its ratio with linoleic acid (LA, C18:2n-6) (LA/ALA) exhibited an effective role as biomarker in discriminating cheese from animals fed fresh forage-based diets. The results showed how dehydrated sulla pellets, as an alternative to hay in periods of limited availability of fresh forage, can preserve the nutritional and health properties of dairy products with regards to their antioxidant capacity and FA profile.

A Multivariate Approach to Study the Bacterial Diversity Associated to the Wooden Shelves Used for Aging Traditional Sicilian Cheeses.

The present study was carried to correlate the microbial diversity of the biofilms developed on the wooden boards used for aging traditional Sicilian cheeses with cheese typology. To this end, the microbial diversity of the shelves in contact with the cheeses PDO Pecorino Siciliano, PDO Piacentinu Ennese, and TAP Caciocavallo Palermitano, during ripening, was evaluated by a multivariate statistical approach. The shelf biofilms of this study were previously analyzed for their microbial composition, but no correlation between biodiversity and cheese type was investigated. Canonical discriminant analysis confirmed a cheese typology effect on the microbial loads of the wooden shelves investigated. Regarding the plate count data, the centroids of different cheeses were statistically distant from one another. This analysis also showed a good graphic separation of data regarding bacterial order operational taxonomy units (OTUs). Thus, the microbiological differences imputed to the cheese typologies were not affected by the environmental conditions of the facilities. Furthermore, wooden shelf lactic acid bacteria (LAB) were investigated for their ability to inhibit the main dairy pathogens. Although inhibitors were mainly enterococci, P. pentosaceus WS287 and W. paramesenteroides WS581 showed the highest inhibition activity, indicating their possible application to control the undesired bacteria in situ.

The Quality of Five Natural, Historical Italian Cheeses Produced in Different Months: Gross Composition, Fat-Soluble Vitamins, Fatty Acids, Total Phenols, Antioxidant Capacity, and Health Index.

Five natural historic cheeses of Southern Italy were investigated-Caciocavallo Palermitano (CP), Casizolu del Montiferru (CdM), Vastedda della Valle del Belìce (VVB), Pecorino Siciliano (PS), and Caprino Nicastrese (CN)-which are produced with raw milk and with traditional techniques and tools, from autochthonous breeds reared under an extensive system. The effects of the month of production on gross composition, MUFA, PUFA, PUFA-ω6, PUFA-ω3, α-tocopherol, retinol, cholesterol, TPC, TEAC, and GHIC were evaluated. In CP, CLA, TPC, and GHIC were higher in April than in February. CdM showed higher values in terms of fat, saturated fatty acids, PUFA-ω3, α-tocopherol, TEAC, and GHIC in May than in February and September, while low values in terms of protein, moisture, and CLA were found. In VVB, MUFA, PUFA-ω6, and α-tocopherol increased in June compared with April; conversely, protein, FRAP, and TEAC were higher in April. In PS, protein, CLA, PUFA, PUFA-ω3, α-tocopherol, and GHIC increased in May compared with January; on the contrary, moisture, NaCl, and TEAC showed high values in January. CN showed higher values in terms of PUFA, PUFA-ω6, PUFA-ω3, TPC, TEAC, and GHIC in April and June compared with January. It is shown that each cheese is unique and closely linked to the production area. Cheeses produced in the spring months showed a high nutritional quality due to the greatest presence of healthy compounds originating from an extensive feeding system.

Amaranthus grain as a new ingredient in diets for dairy cows: productive, qualitative, and in vitro fermentation traits.

BACKGROUND: In recent decades, grain amaranths have attracted attention due to their valuable combination of nutritional traits, with higher protein and oil content than conventional cereals. Before they can be proposed as an unconventional ingredient in animal feed, many aspects still need to be investigated from field production to nutritive value. The present research aimed to study the agronomic traits, proximate composition, and digestibility/degradability, fatty acid profile, antioxidant activity, and total phenolic content of two grain amaranth species, Amaranthus cruentus and Amaranthus hypochondriacus (for a total of six accessions), grown in a Mediterranean environment. RESULTS: Both species showed seed yields comparable to or higher than the traditional cereal crops in the same environment. On the whole, A. cruentus resulted in a higher seed production than A. hypochondriacus. Mexico and Montana accessions, both belonging to A. cruentus, showed the highest yield (3.73 t ha-1 , on average). Few differences emerged in nutritive value between species and accessions: the Illinois accession of A. cruentus showed the best performance in terms of in vitro degradability and gas production, but not for volatile fatty acid production; the fermentation kinetics was slowest in the Illinois accession and fastest in the Montana accession of A. cruentus and the India accession of A. hypochondriacus. CONCLUSION: From a health perspective, the Nebraska accession of A. hypochondriacus represents the best accession, with the lowest saturated fatty acid content and the highest polyunsaturated fatty acid content. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

In-Depth Investigation of the Safety of Wooden Shelves Used for Traditional Cheese Ripening.

The main goal of this research was to characterize the bacterial diversity of the wooden boards used for aging traditional Sicilian cheeses and to evaluate whether pathogenic bacteria are associated with these surfaces. Eighteen cheese dairy factories producing three traditional cheese typologies (PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano) were selected within the region of Sicily. The wooden shelf surfaces were sampled by a destructive method to detach wood splinters as well as by a nondestructive brushing to collect microbial cells. Scanning electron microscopy showed the presence of almost continuous bacterial formations on the majority of the shelves analyzed. Yeasts and fungal hyphae were also visualized, indicating the complexity of the plank communities. The amplicon library of the 16S rRNA gene V3-V4 region was paired-end sequenced using the Illumina MiSeq system, allowing the identification of 14 phyla, 32 classes, 52 orders, 93 families, and 137 genera. Staphylococcus equorum was identified from all wooden surfaces, with a maximum abundance of 64.75%. Among cheese-surface-ripening bacteria, Brevibacterium and Corynebacterium were detected in almost all samples. Several halophilic (Halomonas, Tetragenococcus halophilus, Chromohalobacter, Salimicrobium, Marinococcus, Salegentibacter, Haererehalobacter, Marinobacter, and Idiomarinaceae) and moderately halophilic (Salinicoccus, Psychrobacter, and Salinisphaera) bacteria were frequently identified. Lactic acid bacteria (LAB) were present at low percentages in the genera Leuconostoc, Lactococcus, Lactobacillus, Pediococcus, and Streptococcus. The levels of viable microorganisms on the wooden shelves ranged between 2.4 and 7.8 log CFU/cm2. In some cases, LAB were counted at very high levels (8.2 log CFU/cm2). Members of the Enterobacteriaceae family were detected in a viable state for only six samples. Coagulase-positive staphylococci, Salmonella spp., and Listeria monocytogenes were not detected. Seventy-five strains belonged to the genera Leuconostoc, Lactococcus, Pediococcus, Enterococcus, Lactobacillus, and Weissella. IMPORTANCE This study provides evidence for the lack of pathogenic bacteria on the wooden shelves used to ripen internal bacterially ripened semihard and hard cheeses produced in Sicily. These three cheeses are not inoculated on their surfaces, and surface ripening is not considered to occur or, at least, does not occur at the same extent as surface-inoculated smear cheeses. Several bacterial groups identified from the wooden shelves are typically associated with smear cheeses, strongly suggesting that PDO Pecorino Siciliano, PDO Piacentinu Ennese, and Caciocavallo Palermitano cheese rind contributes to their final organoleptic profiles.

Alkaline Phosphatase Survey in Pecorino Siciliano PDO Cheese.

The determination of alkaline phosphatase (ALP) in cheeses has become an official method for controlling cheeses with a protected designation of origin (PDO), all of which use raw milk. PDO cheeses, characterized by high craftsmanship, usually have an uneven quality. However, for these cheeses, it is necessary to establish ALP values so that they can be defined as a raw milk product. In this study, a dataset with Pecorino Siciliano PDO samples was analyzed to determine ALP both at the core and under the rind. The results showed that there was no significant difference between the different zones in Pecorino cheese. A second dataset of 100 pecorino cheese samples determined that ALP was only at the core of the cheese. Moreover, there was a statistically significant difference between the ALP values of cheeses produced with raw milk and those produced with pasteurized milk. Furthermore, according to the temperatures, a wide variability of ALP values was observed in the Pecorino Siciliano PDO samples from the core of the cheeses. This was a result of several under scotta whey cooking methodologies adopted by cheesemakers, which do not permit a clear range. Therefore, further investigation is desirable.

Improvement of Raw Milk Cheese Hygiene through the Selection of Starter and Non-Starter Lactic Acid Bacteria: The Successful Case of PDO Pecorino Siciliano Cheese.

This review article focuses on the technological aspects and microbiological critical points of pressed-cooked cheeses processed from raw ewe's milk without the inoculation of starter cultures, in particular "Pecorino" cheese typology produced in Italy. After showing the composition of the biofilms adhering to the surface of the traditional dairy equipment (mainly wooden vat used to collect milk) and the microbiological characteristics of PDO Pecorino Siciliano cheese manufactured throughout Sicily, this cheese is taken as a case study to develop a strategy to improve its hygienic and safety characteristics. Basically, the natural lactic acid bacterial populations of fresh and ripened cheeses were characterized to select an autochthonous starter and non-starter cultures to stabilize the microbial community of PDO Pecorino Siciliano cheese. These bacteria were applied at a small scale level to prove their in situ efficacy, and finally introduced within the consortium for protection and promotion of this cheese to disseminate their performances to all dairy factories. The innovation in PDO Pecorino Siciliano cheese production was proven to be respectful of the traditional protocol, the final cheeses preserved their typicality, and the general cheese safety was improved. An overview of the future research prospects is also reported.

Silage of Prickly Pears (Opuntia spp.) Juice By-Products.

Cactus pear cladodes are used as forage in the most arid regions. In Italy, the human consumption of prickly pear fruits and juice is gradually increasing for their numerous health benefits. In manufacturing plants that produce prickly pear juice, several by-products (prickly pear by-products PPB) are obtained. Despite their interesting nutritional characteristics, PPB are not very usable because of their poor shelf-life which is related to their high moisture and sugar content. The aim of this study was to verify the efficacy of ensilage to preserve PPB and to compare different inclusion levels (0, 5, and 10% as fed) of wheat straw. For each treatment, four under vacuum micro-silos were prepared and, after 35 days of storage, the state of preservation was evaluated. Subsequently, the aliquots were analyzed for chemical composition and incubated with bovine rumen fluid to evaluate the fermentation kinetics. The PPB 5% of straw showed significant lower pH and ammonia nitrogen concentration, indicating a better preservation process. Moreover, PPB 5% of straw showed better nutritional parameters (higher crude protein and lower Neutral Detergent Fibre) and fermentation characteristics (higher degradability and VFA volatile fatty acids production) when compared with the other PPB silages. Ensilage with straw represents a suitable storage technique to preserve the nutritional characteristics of PPB.

Addition of selected starter/non-starter lactic acid bacterial inoculums to stabilise PDO Pecorino Siciliano cheese production.

The present study was carried out to produce Protected Denomination of Origin (PDO) Pecorino Siciliano cheese with a multi-species lactic acid bacteria (LAB) culture, composed of starter and non-starter strains in order to reduce the microbiological variability of the products derived without LAB inoculums. To this end, cheese samples produced in six factories located in five provinces (Agrigento, Catania, Enna, Palermo and Trapani) of Sicily, and previously characterised for physicochemical, microbiological and sensory aspects, have been investigated in this work for bacterial microbiome, fatty acid (FA) composition as well as volatile organic compound (VOC) profiles. Analysis of the cheese microbiomes indicated that streptococci (30.62-77.18% relative abundance) and lactobacilli (on average 25.90% relative abundance) dominated the bacterial communities of control cheeses, produced without exogenous inoculums, whereas the cheeses produced with the selected multi-strain culture saw the dominance of lactococci (in the range 6.49-14.92% relative abundance), streptococci and lactobacilli. After the addition of the selected mixed culture, Shannon index increased in all cheeses, but only the cheeses produced with the selected LAB mixed culture in the factory 2 showed Gini-Simpson diversity index (0.79) closer to the reference value (0.94) for a perfect even community. FA composition, mainly represented by saturated FA (on average 69.60% and 69.39% in control cheeses and experimental cheeses, respectively), was not affected by adding LAB culture. The presence of polyunsaturated FA ranged between 7.93 and 8.03% of FA. VOC profiles were different only for the content of butanoic acid, registered for the experimental cheeses at higher concentrations (on average 662.54 mg/kg) than control cheeses (barely 11.96 mg/kg). This study validated addition of the ad hoc starter/non-starter culture for PDO Pecorino cheese production.

Prickly Pear By-Product in the Feeding of Livestock Ruminants: Preliminary Investigation.

In Sicily, the current increasing cultivation of Opuntia ficus-indica corresponds to an availability of prickly pear by-product (PPB) that results from fruit processing for juice extraction. This investigation aims to evaluate the nutritional traits of PPB for ruminant feeding and its stability during a 21-day outdoor storage, using potassium metabisulfite (PMB) as a preservative agent, added to the PPB mass at different doses (0, 50, 100, and 150 g/kg). The fractioning of PPB showed that it included 28% of peel and pulp and 72% of seeds on a dry matter (DM) basis. On the whole, this by-product was low in crude protein (5.32% DM), high in fiber content (51.38%, 41.15% and 14.64% DM for NDFom, ADFom and ADL respectively), non-fiber carbohydrates (NFC, 29.68% DM), and soluble sugars (13.3% DM), with a moderate level of net energy for lactation (4.59 MJ/kg DM). Storage was the main factor of alteration of PPB chemical composition with the exception of ether extract. A decline of NFC and soluble sugars, due to microbial fermentation, was observed with all PMB treatments, especially during the first week of storage, probably due to evolution of both coccus (M17) and rod LAB (MRS), which increased their loads at the seventh day of storage.

Effects of adding solid and molten chocolate on the physicochemical, antioxidant, microbiological, and sensory properties of ewe's milk cheese.

A novel dairy product, namely "chocolate cheese", was produced with two typical Sicilian food products: Pecorino cheese, processed from ewe's milk, and Modica chocolate. The cheese, manufactured with 0%, 5%, 10%, and 15% (w/w) solid or molten chocolate, was evaluated after 0, 2, 4, and 6 weeks of vacuum storage for its nutritional and health properties. The addition of chocolate reduced the pH, protein, fat, and ash; the addition of 5% or 10% molten chocolate reduced hardness (N/mm2 ). The addition of either solid or molten chocolate resulted in a slight increase (P < 0.1038) in the total polyphenol content, a higher oleic acid content, and less oxidative stability. The microbiological profile showed that the total mesophilic count and the number of mesophilic coccus lactic acid bacteria (LAB) were approximately equal (about 108 CFU/g) in all cheese. The survival of the microorganisms was affected by both the chocolate added and the storage time. Chocolate cheese stored for 6 weeks had less Enterobacteriaceae than control cheese, whereas yeasts were detected at higher cell densities in the former cheese. Filamentous fungi were undetectable in some cheese. Differences were also observed in the number of mesophilic rod LAB, which increased progressively over time in all cheese, and in Enterobacteriaceae, yeasts, and filamentous fungi, which decreased during storage. Descriptive and hedonic sensory tests and principal component analysis showed that fresh cheese and cheese stored for 2 weeks, including 5% molten chocolate, were the most preferred by evaluators. Based on these results, chocolate cheese has the potential to be appreciated in the market for its nutritional, health, and sensory properties. PRACTICAL APPLICATION: Chocolate cheese, made by combining two typical Sicilian foods, Pecorino cheese and Modica chocolate, is proposed as a novel dairy product. The highest sensory acceptance was obtained with the addition of 5% molten chocolate and storage for 2 weeks. Given its improved antioxidant properties, healthier fat, and sensory properties, chocolate cheese has the potential to be appreciated in the market, especially by young consumers.

Improvement of Oxidative Status, Milk and Cheese Production, and Food Sustainability Indexes by Addition of Durum Wheat Bran to Dairy Cows' Diet.

Durum wheat bran (DWB) is a by-product mostly used in feeding ruminants, contributing to decrease in the utilization of feeds suitable as foods for human consumption, thus improving the sustainability of livestock production. However, the potential benefits of DWB, due to its content in phenolic acids, mainly consisting of ferulic acid with antioxidant properties, have not been well clarified yet. Accordingly, in this experiment, 36 lactating cows divided into three groups received, over a period of 100 days, one of three concentrates including DWB at 0% (DWB0), 10% (DWB10), or 20% (DWB20). The concentrates were formulated to be isoproteic and isoenergetic and, to balance the higher fiber content of the concentrates with DWB, the hay in the diets was slightly reduced. During the trial, the group feed intake and the individual milk production were monitored, and cheese was made with bulk milk from each group. Milk yield and microbiological characteristics of milk and cheese were similar among groups, indicating no DWB effect on cows performance and fermentation process. Milk from DWB20 group resulted slightly higher in casein and curd firmness (a2r). In cows fed DWB, the higher polyphenol intake was responsible for higher blood contents of these bioactive compounds, that seemed to have contributed in reducing the level of reactive oxygen metabolites (ROMs), which were higher in DWB0 cows. DWB20 cheeses showed a higher polyphenol content, lower number of peroxides, and higher antioxidant capacity than DWB0 cheeses. DWB20 and DWB10 diets resulted less expensive. In addition, the DWB20 group showed the best indexes heFCE (human edible feed conversion efficiency = milk/human edible feed) and NFP (net food production = milk - human edible food), expressed as crude protein or gross energy. In conclusion, the DWB fed to dairy cows at 12% of diet dry matter (DM) can lead to benefits, such as the improvement of oxidative status of cows, milk quality, shelf-life, and functional properties of cheese, and might contribute to reduce the feeding cost and limit the human-animal competition for feeding sources.

Transformation of raw ewes' milk applying "Grana" type pressed cheese technology: Development of extra-hard "Gran Ovino" cheese.

This work was carried out to pursue a double objective: to improve the hygienic safety of cheeses produced from raw ewes' milk; and to produce a new typology of raw ewes' milk through the application of "Grana" technology for which the name "Gran Ovino" was chosen. With this in mind, raw milk from an individual farm was transformed under controlled conditions at a dairy pilot plant. The production technology included the partial skimming of the evening and morning milk mixture by cream surfacing and the addition of a natural whey starter cultures (NWSC) prepared with four selected Streptococcus thermophilus strains (PON6, PON244, PON261 e PON413). Ten microbial groups were investigated by plate counts from raw milk until ripened cheeses. Lactic acid bacteria (LAB) were in the range 104-105 CFU/ml before NWSC addition. After curdling, this group increased by 3 log cycles and was counted at 106 CFU/g after curd cooking. A rapid pH drop (to 6.05) was registered after almost 3 h from NWSC addition. The levels of members of the Enterobacteriaceae family were at about 103 CFU/ml in raw milk and decreased after curd cooking to 1 log cycle. A similar behavior was shown by the other undesired microbial groups and a complete disappearance of staphylococci was registered. The microbiological counts of 9-month ripened cheeses showed the dominance of LAB and undetectable levels of the undesired bacteria. MiSeq Illumina was applied to better investigate the bacterial composition of ripened cheeses and this technique evidenced that the majority of OTUs belonged to Lactobacillus and Streptococcus genera. The final cheeses were characterized by 67.65% dry matter of which 41.85% of fats and 47.02% of proteins. The main cheese fatty acids were palmitic, oleic and myristic acids and the saturated fatty acids/unsaturated fatty acids ratio was 2.17. Forty-one volatile compounds, including acids, esters, ketones, alcohols, aldehydes, phenols and one terpene were emitted from the cheese. Sensory evaluation showed a general appreciation for the new cheese product by judges.

Effects of Diets Supplemented with Medicinal Mushroom Myceliated Grains on Some Production, Health, and Oxidation Traits of Dairy Ewes.

The beneficial properties of mushrooms' bioactive compounds indicate their potential for use as performance-enhancing natural additives for livestock animals. A study was undertaken to evaluate the effects of diets supplemented with mushroom myceliated grains (MMGs) fed to dairy ewes on intestinal parasite load, milk production, milk fatty acid (FA) composition, and cheese oxidative stability. During an 8-week experimental period, 21 lactating Valle del Belice ewes were divided into 3 groups named MMG20, MMG10, and MMG0. Ewes in each group were fed hay ad libitum and 1.3 kg/day/head of 1 of 3 concentrates with MMGs at 20% (MMG20), 10% (MMG10), or 0% (MMG0). The ewes fed MMG20 had comparable dry matter (DM) and nutrients intake, fewer intestinal parasite infections, a tendency toward higher milk yield, and higher milk casein content (4.78% in MMG20 vs. 4.32% in MMG10 and 4.27% in MMG0; P < 0.05), and they produced cheese with less intense yellow color and a lower secondary lipid oxidation, than the ewes in the MMG10 and MMG0 groups. A higher antioxidant capacity was observed (17.83 mmol Trolox equivalent/kg DM in the MMG20 group vs. 9.97 and 9.18 mmol Trolox equivalent/kg DM in the MMG10 and MMG0 groups, respectively; P < 0.001), suggesting a higher oxidative stability of cheese fat and a probable enrichment of cheese with antioxidant compounds inherent in or induced by MMGs. The inclusion of MMGs in the diet did not affect the amounts of health-promoting polyunsaturated FAs in milk, with the exception of n-3 eicosapentaenoic acid, which was found only in milk from the MMG-treated ewes. These promising results merit further investigation into the potential use of medicinal mushrooms to enhance animal health and production.

Influence of the early bacterial biofilms developed on vats made with seven wood types on PDO Vastedda della valle del Belìce cheese characteristics.

Early vat bacterial biofilms developed spontaneously through contact with whey have been characterized on seven wood types (Castanea sativa Miller, Cedrus libani, A. Rich., Prunus avium L., Fraxinus ornus L., Juglans regia L., Pinus nigra J.F. Arnold and Populus nigra L.). The present study aimed to evaluate the influence of these biofilms on the microbiological, chemical, physical and sensory characteristics of PDO Vastedda della valle del Belìce (VdB) cheese, processed traditionally from raw ewe's milk using wooden tools. To this purpose, the experimental cheeses after 15 d of refrigerated storage were examined. Lactic acid bacteria (LAB) populations dominated the microbial community of all samples. The species more frequently identified were Lactococcus lactis among starter LAB and Lactobacillus paracasei, Lactobacillus rhamnosus, Lactobacillus fermentum and Pediococcus pentosaceus among non starter LAB. Culture-independent analysis of microbiota diversity was performed by MiSeq Illumina that identified Streptococcus as major group followed by members of Enterobacteriaceae family, Lactococcus and Lactobacillus. Generally, the seven tree species did not negatively affect the physicochemical composition of VdB cheeses. Chestnut (both Sicilian and Calabrian) vats produced cheeses with significant lower hue angle (a*/b*) than other wood types. Among chemical parameters, significant variations were registered for aw, primary and secondary lipid oxidation state (significantly lower for the VdB cheeses produced with poplar wood), and volatile organic compounds (VOCs). The significant differences detected among the VOCs emitted from cheeses were not perceived by the panelists who recognized all cheeses from the different trials as similar. This study confirmed the suitability of cedar, cherry, ash, walnut, black pine and poplar as alternative woods to chestnut for the production of the wooden vats employed in cheese making for the Sicilian traditional dairy productions.

Retrospective study on the hygienic quality of fresh ricotta cheeses produced in Sicily, Italy.

The present work was carried out to investigate the microbiological profile of Sicilian ewes' ricotta cheeses during fifteen years of investigations (2002-2016). The samples were collected between those conferred to the Istituto Zooprofilattico Sperimentale della Sicilia (IZSSi) Adelmo Mirri, Palermo (Italy), by the competent authority during official control, by food business operator in HACCP systems and in research projects. Enterobacteriaceae, Escherichia coli and coagulase-positive staphylococci (CPS) were found only in some samples. Bacillus cereus was detected in c.a. 16% of samples but the level of contaminations did not reach the threshold that leads to significant toxin production. Pathogenic bacteria such as Listeria monocytogenes, Salmonella spp. and Brucella spp. were never detected. Furthermore, a total of 47 of lactic acid bacteria (LAB) strains were identified at species level by sequencing the 16S rRNA gene, resulting in the identification of 10 species belonging to four genera Enterococcus, Lactobacillus, Lactococcus and Leuconostoc, commonly employed as starter and non starter cultures in different traditional cheese. Results of this study highlighted an improvement of the hygienic conditions of the Sicilian ewes' ricotta cheeses during the last ten years of investigation. This observation was confirmed from reduction of undesired microorganisms such as Enterobacteriaceae, E.coli and CPS, used to define the process hygiene criteria. However, in order to improve the final quality of this product are needed further strategy such as the dairy makers training, with the aim to apply a good hygienic practices during the production.