The profile of phenolic compounds changes during the growth of a plant and this change affects its antioxidant potential. The aim of this research has been to find the growth stage of flax with the highest antioxidant capacity, and to determine the phenolic compounds responsible for such a capacity. Flax was harvested in six growth stages: from stem extension to mature seeds. The phenolic compounds were identified using LC-TOF-MS and quantified in an extract and in the fresh matter (FM) of each growth stage. The radical scavenging activity against ABTS and DPPH, the ferric-reducing antioxidant power (FRAP), and the antioxidant activity in the beta-carotene-linoleic acid emulsion system were determined. Mono- and di-C-glycosyl flavones were found to be the most abundant phenolics of the aerial parts of flax, which also showed the highest content of isoorientin (210-538 µg/g FM). Coniferin, its derivative, and hydroxycinnamic acid derivatives were also detected. The plant was richer in flavone C-glycosides from stem extension to seed ripening (1105-1413 µg/g FM) than at the mature seed stage (557 µg/g FM). Most of the individual flavone C-glycoside contents in the extracts decreased when increasingly older plants were considered; however, the isoorientin content did not change significantly from the steam extension to the seed ripening stages. The antiradical activity against ABTS and FRAP was higher for the aerial parts of the flax harvested at the flowering, brown capsule, and seed ripening stages, mainly due to the presence of flavone C-glycosides. The oxidation of beta-carotene-linoleic acid emulsion was instead inhibited more effectively by the extracts from plants at the brown capsule and mature seed stages. Coniferin and its derivative were significantly involved in this activity. The extracts from the aerial parts of the flax harvested from flowering to seed ripening could be a valuable source of flavone C-glycosides for use as nutraceuticals and components of functional foods.

Currently, there is a renewed interest in cannabis-related products in different fields be-cause of the rich phytocomplex of this plant, together with its fiber and agricultural features. In this context, the current study aims to chemically characterize different samples of fiber-type Cannabis sativa L. grown in Italy as a potential health promoting source. An ultrasound-assisted solid-liquid extraction (UA-SLE) method was first developed and optimized to obtain a fingerprinting of the investigated phytocomplex. Analyses were carried out through an ultra high performance liquid chromatography equipped with a photodiode array detector in series with triple quadrupole system with an electrospray ionization (ESI) interface (UHPLC-UV-ESI-MS/MS) and showed that the phytocomplex mainly includes flavonoids and non-psychotomimetic cannabinoids. The method was then applied to characterize and compare 24 samples of fiber-type Cannabis sativa L. aerial parts (mainly stems and leaves), which differed for the growth stages (from mid-vegetative to early flowering), growth land plots, and methods of drying (forced-draft oven or freeze-drying). The quali-quantitative analysis showed that a freeze-drying method seems to better preserve the chemical composition of the samples, while the location of the land plot and the growth stage of the plant (which did not comprise inflorescences) had minor influences on the chemical pattern. These results were also supported by spectrophotometric in-vitro assays (scavenging of 2,2-diphenyl-1-picrylhy-drazyl (DPPH) and 2,2?-azinobis-3-ethyl-benzthiazoline-6-sulphonate (ABTS) radicals and inhibitory activity against tyrosinase and elastase enzymes) to investigate the potential biological activity of these samples and the contribution of non-psychotomimetic cannabinoids.

Traditional foods are gaining more and more market due to consumers' increasing willingness to buy products linked to national cultures: among these products, cheese plays an important role. Plaisentif is a traditional Piedmont cheese, only made during violets blooming season. The aim of this work is to evaluate the safety of this cheese, taking into account the EU Regulations. Microbiological hazards as well chemical, biogenic amines and mycotoxins, analysis were investigated. Salmonella spp. and Listeria monocytogenes were never detected in cheeses after ripening. Biogenic amines were present in very low quantities. Ochratoxin A was never detected and patulin was detected in over one cheese during the two years of sampling. This is the first attempt to characterize traditional Plaisentif cheese from a safety point of view. All the information acquired can be held as a necessary basis for reinforcing the culture of traditional products, for economic opportunities in mountainous regions and for safeguarding traditions and cultural identities.

Fresh mechanically deboned meat (MDM) is usually claimed as high-quality ingredient on dry pet food recipes and this aspect may positively influence consumer choice. It is important to determine the scientifically sustainability of this claim and to assess the microbiological safety of MDM inclusion in dry pet food. Objectives were: 1) to evaluate the effect of inclusion of MDM in dry dog food on fatty acid profile and in vivo and in vitro digestibility, proposing a new system (DaisyII Incubator) to measure the in vitro digestibility for dogs; 2) to compare palatability of dry dog food containing MDM with dry dog food in which meat by-products (MBP) are the only animal protein sources; 3) to determine, whether or not, the inclusion of that ingredient changes the microbiology and the storage quality. Results indicated that MDM product was characterized by significant higher nutritional value in terms of fatty acids profile, in vitro digestibility (HV-IVD method) and lower palatability than the MBP product. Microbiological risk assessment showed no microbiological hazards for either product. After 6-months storage, the total mesophilic bacterial count ranged between 1.77 and 2.09 log CFU/g feed, while polyamine values were higher in the MDM (0.37 g/kg) than in the MBP (0.27 g/kg). The DaisyII Incubator was found to be a valid instrument for studying in vitro digestibility also for dogs, providing data simply, quickly, with less variability and costs than in vivo trials. In conclusion, MDM inclusion in dry dog food is microbiologically safe and it can improve its nutritional quality, at the expense of a reduced palatability. The higher polyamine levels fount in MDM-enriched petfood after 6-months storage, however, may represent a possible hazard, and further studies are still warranted.

The aim of this study was to find a correlation between in vivo NIR scan patterns, oxidative status, and blood serum metabolites in rabbits fed diets protected or unprotected against oxidation. Rabbits does in groups of eight were fed for 9 weeks with diets containing linseed, rich in polyunsaturated fatty acids (LS), or linseed plus hazelnut skins, with antioxidant function (LS+HS), and palm oil, rich in saturated fatty acids (PO). The animals were examined at days 1, 31, and 63 using a smart SCIO (TM) molecular sensor, a new miniaturized web-based wireless device, applied to the internal ear flap (NIR range 740-1070 nm). The hazelnut peels integrated diet protected the rabbits from the oxidative stress induced by the addition of unprotected polyunsaturated fats. NIR tomoscopy was variously correlated with serum parameters, lysozyme (R-2=0.71), ROMs (0.47), cholesterol (0.49), triglycerides (0.40), and a multivariate Index of Oxidative Risk (0.67 (IOR)). The correlations suggested a close connection between the clustering of the diets according to the laboratory variables and the NIRS scan pattern clustering at the ending trial day, as shown by the highly significant odds ratios. Advantageous use of this simple, painless technique was evident in the planning phase, with no difference among the groups at the beginning of the study, but an effect size that evolved differently over time until the end of the study. In a practical validation of the SCIO (TM) model in 92 commercial rabbit does, the average spread of the predicted IOR was 33% in lactating does (2.54 +/- 0.05) vs. dry does (1.91 +/- 0.07).

Two investigations were carried out in the Northwest Italy (Susa and Chisone Valleys). The pastures were natural grasslands without any kind of fertilization and were located in five sites at an altitude range of 1704-2070 m a.s.l. during the first trial in the 2013 grazing season and in nine sites at an altitude range of 1620-2070 m a.s.l. during the second trial in the 2014 grazing season. Collected samples were analysed for total phenol (TP), total tannin (TT) and condensed tannin (CT) contents and results were correlated to chemical composition and in vitro digestibility reported in previous papers. A significant correlation between all the tannin fractions was found regarding the acid detergent lignin (ADL) pasture contents. A higher correlation (P<0.01) was found for TP and TT fractions (0.747 and 0.717, respectively), while CT fraction showed a lower one (0.436, P<0.05). Principal component analysis confirmed that as far as tannins fractions are concerned, TP and TT were closely associated with ADL as shown in the distribution of the variables (positive quadrant of the right part), while CT fraction was less correlated. The results highlight the importance of the tannins quantity and quality of the pasture, that can, due to the changes in the nutritional composition of pastures, affect milk quality during the grazing season.

The aim of this study was to evaluate the differences in the antioxidant activity and phenolic profile of sunflower (Helianthus annuusL.) extracts obtained from the aerial parts of plants harvested at five growth stages. In vitro assays were used to determine the antioxidant activity, i.e., ABTS(center dot+)and DPPH(center dot)scavenging activity, the ferric-reducing antioxidant power (FRAP) and the ability to inhibit beta-carotene-linoleic acid emulsion oxidation. Phenolic compounds, such as mono- and dicaffeoylquinic acid isomers and caffeic acid hexose, were identified using the LC-TOF-MS/MS technique. The predominant compound during the growth cycle of the plant was 3,5-di-O-caffeoylquinic acid, whose content was the highest at the mid-flowering stage. The total phenolic content was also the highest in sunflowers at the mid-flowering stage. The main phenolic compound contents were closely correlated with ABTS(center dot+)and DPPH(center dot)scavenging activity and FRAP. No significant correlation was found between the total phenolic content and the antioxidant activity in the emulsion system. The highest antiradical activity and FRAP were generally determined in older plants (mid-flowering and late flowering stages). In conclusion, the aerial parts of sunflowers, in particular those harvested at the mid-flowering stage, are a good plant material from which to obtain phenolic compound extracts, albeit mainly of one class (esters of caffeic acid and quinic acid), with high antioxidant activity.

The popularity of edible wild fruits has increased in industrialized countries due to their composition and positive effects. The aim of this study has been to characterize the polyphenolics and anthocyanins of black mulberry (Morus nigra L.), cornelian cherry (Cornus mas L.), elderberry (Sambucus nigra L.), hawthorn (Crataegus monogyna L.), lingonberry (Vaccinium vitis-idaea L.) and rose hip (Rosa canina L.) harvested in the north-west of Italy by means of HPLC-DAD-ESI HRMS in positive ion mode. Although there is an abundant amount of literature related to the polyphenolics of cultivated fruit, a new type of comparison has here been conducted between wild and cultivated fruits on their polyphenolic content. The HPLC-DAD-ESI HRMS method has detected 64 different polyphenolic molecules and it can be used to perform qualitative and quantitative analyses. Furthermore, the cornelian cherry and elderberry samples showed the highest polyphenolic compound levels. The quercetin glycosylated compounds showed the highest percentage of flavonols in most of the analyzed wild fruits.

This is a reprint of articles from the Special Issue published online in the open access journal Animals (ISSN 2076-2615) (available at: https://www.mdpi.com/journal/animals/special issues/ In vitro digestibility).

The phenolic compound profile and antioxidant potential of the false flax (Camelina saliva L.) plant, harvested at five morphological stages, that is. from the vegetative to the ripe seed-pod stage, have been investigated. False flax extracts were prepared using 80% (v/v) methanol. and the total phenolic content (TPC), the contents of the individual phenolics and antioxidant activity, measured as the Trolox equivalent antioxidant capacity (TEAC), ferric-reducing antioxidant power (FRAP), DPPH center dot scavenging activity and the ability to inhibit the oxidation of beta-carotene-linoleic acid emulsion, were determined. The TPC of the plant, at different growth stages, ranged from 49.2 to 59.1 mg GAE/g of extract and from 1.46 to 3.10 mg GAE/g of fresh matter (FM). Four main phenolic compounds were identified (chlorogenic acid, rutin, quercetin 3-O-glucoside, and quercetin glycoside). The chlorogenic acid content and the sum of flavonoids increased in the extracts from the vegetative to the bud stage, reaching 35.9 and 49.5 mg/g of extract, respectively, and gradually decreased in the subsequent growth stages. The plant extracts at the bud and flowering stages generally had the highest antioxidant activity in the polar systems (TEAC, FRAP and DPPH assays). The ripe seed-pod stage showed the highest antioxidant potential in these conditions when the results were expressed on FM basis. The best antioxidant activity in the lipid emulsion system was shown for the false flax extracts at the flowering and ripe seed-pod stages. Our research has indicated the possibility of using the aerial part of C. saliva as a source of ingredients with protective antioxidant activity.

The popularity of small berries has rapidly increased in Western countries given their antioxidant, anti-inflammatory, and antimicrobial activities and health-promoting properties. The aim of this study was to compare the fatty acid (FA) profile, phenolic compounds, and antioxidant capacity of extracts of 11 berries cultivated in the North West of Italy. Berry samples were extracted and evaluated for FA profile and total anthocyanin (TAC), total flavonoid contents (TFC), ferric-reducing antioxidant power (FRAP), and for their radical scavenging activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. The main polyphenols of berry extracts were characterized by HPLC-DAD-UV-ESI HRMS in positive ion mode. Results showed that the highest TAC and TFC contents were recorded in black currants, blackberries, and blueberries. Maximum and minimum DPPH radical scavenging activities, Trolox Equivalent Antioxidant Capacity, and FRAP measurements confirmed the same trend recorded for TAC and TFC values. HPLC-HRMS analyses highlight how blueberries and blackberries have the highest concentration in polyphenols. Palmitic, stearic, oleic, linoleic, alpha-linolenic, and alpha-linolenic acids significantly differ between berries, with oleic and alpha-linolenic acid representing the most abundant FAs in raspberries. Among the berries investigated, results of phytochemical characterization suggest choosing black currants and blueberries as an excellent source of natural antioxidants for food and health purposes.

The aim was to evaluate the effect of blueberry pomace (BBP) on the oxidative stability and cooking properties of pork patties (PPs) during chilled storage. PPs were prepared into four groups: control, control with the addition of 0.3% ascorbic acid, with 1% BBP, and with 2% BBP. Three PPs were prepared for each batch and analyzed raw after 1, 3, and 7 days of refrigerated storage, respectively. Another three PPs were cooked and sampled for each batch at the same sampling times. An HPLC-HRMS method and SPME GC-MS techniques were developed to identify and quantify the main polyphenols present in the BBP and the lipid oxidation compounds generated in cooked PPs, respectively. The application of the BBP to PPs resulted in desirable changes in the cooking characteristics of the products. BBP is also able to significantly reduce oxidation volatile compounds concentration with the consequent delay of lipid oxidation. Practical applications: Blueberries have a high polyphenol content and, for commercial purposes, a large quantity of this fruit is transformed into juices, and waste products that are generally discarded are produced. Addition of blueberry pomace (BBP) possessing high antioxidant activity can provide a good way of reducing the rate of oxidation changes in the in ground meat products during storage. Recent research has promoted the use of berry pomace for animal feeding purpose and for the development of novel functional food ingredients. The use of BBP as additives provides some beneficial health effects to meat consumers with a reduction of waste products. The present study demonstrates that the application of the BBP as a food additive to pork patties resulted in desirable changes in the cooking characteristics of the products due to an increase in the water retention capacity of the matrix.

This review summarises the use of the Ankom DaisyII incubator (ADII; Ankom Technology Corporation Fairport, NY, USA), as presented in studies on digestibility, and its extension to other species apart from ruminants, from its introduction until today. This technique has been modified and adapted to allow for different types of investigations to be conducted. Researchers have studied and tested different procedures, and the main sources of variation have been found to be: the inoculum source, sample size, sample preparation, and bag type. In vitro digestibility methods, applied to the ADII incubator, have been reviewed, the precision and accuracy of the method using the ADII incubator have been dealt with, and comparisons with other methods have been made. Moreover, some hypotheses on the possible evolutions of this technology in non-ruminants, including pets, have been described. To date, there are no standardised protocols for the collection, storage, and transportation of rumen fluid or faeces. There is also still a need to standardise the procedures for washing the bags after digestion. Moreover, some performance metrics of the instrument (such as the reliability of the rotation mechanism of the jars) still require improvement.

The aim was to determine the absence of gluten in pet food samples marked as 'grain-free' and 'gluten-free' diets, to assess the reliability of manufacturer labelling of such products. A total of 15 diets labelled as grain- or gluten-free and 2 commercial diets containing wheat were sampled. An analytical procedure using high-pressure liquid chromatography coupled with mass spectrometry with high power of resolution was developed and applied to determine specific markers of wheat gluten. The results are expressed as mg of wheat flour type '00' present in 1 g of feed. The quantification limit (LOQ) obtained in the flour for ion m/z 894.5043, z = 2, is 4 mg of flour per gram. In 14 out of the 15 samples from a grain- or gluten-free diet the quantifier ion signal was < LOQ, while in 1 out of the 15 samples 10 mg of flour/g feed were measured.

This review outlines the current state of knowledge on the nutritive value of black cumin (Nigella sativa L.). The popularity of this plant, which is an annual herbaceous plant belonging to the Ranunculaceae family and is native to Iran, Pakistan and Turkey, is due to its beneficial actions. Black cumin has many therapeutic effects and is considered one of the most important medicinal plants in the world because of its antioxidant, anticoccidial, anthelminthic and antimicrobial activities. The nutritive value of black cumin is result of its carbohydrate, fatty acid, protein contents as well as its several bioactive compounds. The seeds or their byproducts can be used in feeds for farm animals, with positive effects on the compositional characteristics of eggs, milk and meat.

A study was conducted over the summer of 2014 on nine Alpine pastures in the Chisone and Susa Valleys (NW Italy). The aim was to characterize the variation in the chemical composition, gross energy, in vitro true digestibility (IVTD), in vitro neutral detergent fiber digestibility (NDFD), fatty acids (FA), total phenols, total and condensed tannin contents, and terpenoid profile. The dry matter, ash, crude protein, acid detergent fiber, lignin, and gross energy contents of the pastures were found to differ. All the pastures had good IVTD (706-829 g/kg DM) and NDFD (487-694 g/kg NDF) values. The most abundant FAs in all the pastures were ?-linolenic (354-519 g/kg of the total FAs), linoleic (75-110 g/kg of the total FAs), and palmitic acid (64-89 g/kg of the total FAs) and they differed significantly among pastures. No significant differences were found in the total phenols, or in the total and condensed tannin contents among pastures. Fifty-eight terpenoids were detected and 4-cyclopentene-1, 3-dione, ?-caryophyllene, and eucalyptol were the most abundant. The terpenoids differed both qualitatively and quantitatively among pastures. The results highlight the importance of the great biodiversity of pastures, which provide a balanced distribution of fundamental nutritional elements and bioactive compounds in grasslands.

Green pruning residues (GPRs) and leaves from 16 red and white Vitis vinifera L. cultivars from Piedmont (Italy) were studied. The investigated samples were extracted by ultrasound-assisted extraction optimized by an experimental design, and quali- and quantitatively analyzed by HPLC-PDA-MS/MS. GPRs and leaves show a similar polyphenolic pattern, with quercetin 3-O-glucuronide, caftaric acid, and quercetin 3-O-glucoside as the main components, although in variable proportions. The HPLC results were related to the antioxidant activity, measured as total phenolic content and through DPPH and ABTS assays with similar results. Colorimetric in vitro assays, offline combined with HPLC-PDA analysis, determine which compounds contribute to the antioxidant activity in terms of radical scavenging abilities. Valorization of GPRs is a potential source of natural compounds that could be of interest in the health field, increasing their economic value together with a positive effect on the environment.

Vitis vinifera L. (Vitaceae family) is a popular woody perennial plant commonly used in the winemaking. Unfortunately, viticulture generates huge amounts of residues (i.e. leaves, twigs, seeds, skins), which can cause environmental issues when discarded in open areas. It is therefore extremely important to evaluate the potential reuse of these matrices, to reduce the amount of waste at landfill sites [1],[2]. Among the main grapevine by-products, no data on the green pruning residues (GPRs) produced by the annual spring pruning have been reported. Thus, the aim of this work was the analysis and valorization of GPRs from 16 red and white V. vinifera cultivars from Piedmont (Italy), by comparing the results with those obtained from the leaves of the same cultivars. An experimental design optimization was performed and the obtained hydroalcoholic extracts where then quali- and quantitatively analyzed by HPLC-PDA-MS/MS, with similar polyphenolic profiles for both GPRs and leaves. Quercetin 3-O-glucuronide, caftaric acid and quercetin 3-O-glucoside were the main components of the investigated matrices, although in variable proportions. Considering the prevalent presence of polyphenols in the analyzed extracts their antioxidant activity was evaluated with colorimetric in-vitro assays, off-line combined with HPLC-PDA analysis. In this way the contribution of each compound to the antioxidant activity, in terms of radical scavenging abilities, was determined. These findings could suggest GPRs as a potential source of natural compounds, and therefore promote their use in the food field, as food supplements, which could increase their economic value together with a positive effect on the environment.

Interest in the green pruning residues of grapevine (GPR), harvested in spring, and in grapevine leaves (GL), harvested in autumn, as a feedstuff, has been increasing due to their nutritive value. The aim of this study has been to investigate the differences between the chemical composition, gross energy (GE) and in vitro apparent digestibility (DMD) of the GPR and GL of five varieties of red grapevine (Cabernet franc, Canaiolo nero, Carignan noir, Lambrusco salamino, and Sangiovese) and of five varieties of white grapevine (Malvasia bianca di Candia, Moscato bianco, Sauvignon blanc, Verdicchio and Vernaccia di S. Gimignano). The dry matter, acid and neutral detergent fibre, GE and DMD were found to differ (P < 0.01) between cultivars for both GPR and GL, while no significant differences were observed between the cultivars, in terms of their mean acid detergent lignin content for GPR and crude protein content for GL, respectively. In conclusion, the results show that both the GL and GPR obtained from red and white grape varieties originating from Italy are suitable dietary sources for ruminant feeding, even though GL has a lower fibrous content than GPR and, consequently, a higher DMD.