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The molecular approach of DNA barcoding for the characterization and traceability of food products has come into common use in many European countries. However, it is important to address and solve technical and scientific issues such as the efficiency of the barcode sequences and DNA extraction methods to be able to analyze all the products that the food sector offers. The goal of this study is to collect the most defrauded and common food products and identify better workflows for species identification. A total of 212 specimens were collected in collaboration with 38 companies belonging to 5 different fields: seafood, botanicals, agrifood, spices, and probiotics. For all the typologies of specimens, the most suitable workflow was defined, and three species-specific primer pairs for fish were also designed. Results showed that 21.2% of the analyzed products were defrauded. A total of 88.2% of specimens were correctly identified by DNA barcoding analysis. Botanicals (28.8%) have the highest number of non-conformances, followed by spices (28.5%), agrifood (23.5%), seafood (11.4%), and probiotics (7.7%). DNA barcoding and mini-barcoding are confirmed as fast and reliable methods for ensuring quality and safety in the food field.

Nowadays, food authentication is more and more required given its relevance in terms of quality and safety. The seafood market is heavily affected by mislabelling and fraudulent substitutions/adulterations, especially for processed food products such as canned food items, due to the loss of morphological features. This study aims to develop new assays based on DNA to identify fresh mackerel (Scomber spp.) and commercial products. A new primer pair was de novo designed on the 5S rRNA gene and non-transcribed spacer (NTS), identifying a DNA mini-barcoding region suitable for species identification of processed commercial products. Moreover, to offer a fast and low-cost analysis, a new assay based on recombinase polymerase amplification (RPA) was developed for the identification of fresh ‘Sgombro’ (Scomber scombrus) and ‘Lanzardo o Occhione’ (Scomber japonicus and Scomber colias), coupled with the lateral flow visualisation for the most expensive species (Scomber scombrus) identification. This innovative portable assay has great potential for supply chain traceability in the seafood market.

Several bamboo species, suitable for temperate climates, have been introduced from southern China as ornamental plants in Europe and in Italy since the XIX century. Bamboos were appreciated also for other commercial purposes afterwards (e.g., food and structural applications) and nowadays in NW Italy (Piedmont and Aosta Valley Regions) the interest in their plantation has been growing. Consequently, many public authorities responsible for environment and biodiversity protection expressed a precautionary attitude for the commerce of bamboos. ‘BambApp’ project was implemented to scrutinize the invasive potential of naturalized bamboo species with a ‘citizen science’ approach, which implied the support of volunteers to collect scientific information through the use of “iNaturalist” app for smartphones. Citizens described each recorded bamboo clump in terms of maximum height, maximum stem diameter, and clump size. The coordinates and a description of the habitat type on the border of the four sides for each clump were required to volunteers as well. Moreover, surveyors had to upload four pictures of the clump, which were used to identify the bamboos at species level. In a ten-month period, 871 validated bamboo populations were recorded in the two regions and nine different species identified. The most widespread species was Phyllostachys aurea (67%), followed by Phyllostachis viridiglaucescens (16%), Pseudosasa japonica (6%), Phyllostachys reticulata (5%), Phyllostachys nigra (2%), Phyllostachys flexuosa (2%), Phyllostachys sulfurea (2%), and Phyllostachys edulis (1%). To quantify the invasive potential of bamboo species a Principal Component Analysis was carried out by using variables related to clump growth. Average elevation ranged between 252 m and 431 m a.s.l. and the clump size ranged between an average of about 27 m2 and about 2160 m2. Phyllostachis reticulata and P. sulfurea were the tallest species with the largest stem diameters, whereas P. aurea was the speci

Hepatic-related diseases, in particular hyperlipidemia and hypercholesterolemia, are a thorn on the side of the national health institutes around the globe. Indeed, liver lipid and cholesterol dysregulation could lead to atherosclerotic plaque formation and cardiovascular diseases. Currently, statin administration and monacolin K consumption are the main therapies proposed to counter this alarming connection, but relevant side effects are known. To overcome this issue, safe nutraceutical formulations and/or vegetal extracts, endowed with anticholesterolemic activity, could be instrumental in hypercholesterolemia prevention and treatment. In the present work, the anticholesterolemic efficacy of three vegetal extracts used in traditional medicine (artichoke, caigua, and fenugreek), their unique blend (ACFB), and the monacolin K-containing red yeast extract (RYR), was investigated with an in vitro approach based on hepatic cell line HepG2. The impact on cholesterol of the three extracts, their blend, and RYR were investigated by determining hepatocyte total and free cholesterol and bile acids biosynthesis. According to our results, the anticholesterolemic activity of the vegetal extracts was confirmed, and a novel choleretic activity of caigua extract was evidenced. ACFB showed to be safer than RYR while showing a similar effect on total and free cholesterol and bile acids synthesis compared to it. The anticholesterolemic activity of the blend was obtained with lower vegetal extract concentrations compared with the single vegetal extract, potentially indicating an additive effect between the extracts. In conclusion, the vegetal extracts and their blend, ACFB, are safe and are endowed with anticholesterolemic activity, potentially providing complementary therapies to the statin-based ones for hyperlipidemia and hypercholesterolemia-related complications.

Medicinal plants have been widely used in traditional medicine due to their therapeutic properties. Although they are mostly used as herbal infusion and tincture, employment as ingredients of food supplements is increasing. However, fraud and adulteration are widespread issues. In our study, we aimed at evaluating DNA metabarcoding as a tool to identify product composition. In order to accomplish this, we analyzed fifteen commercial products with DNA metabarcoding, using two barcode regions: psbA‐trnH and ITS2. Results showed that on average, 70% (44–100) of the declared ingredients have been identified. The ITS2 marker appears to identify more species (n = 60) than psbA‐trnH (n = 35), with an ingredients’ identification rate of 52% versus 45%, respectively. Some species are identified only by one marker rather than the other. Additionally, in order to evaluate the quantitative ability of high‐throughput sequencing (HTS) to compare the plant component to the corresponding assigned sequences, in the laboratory, we created six mock mixtures of plants starting both from biomass and gDNA. Our analysis also supports the application of DNA metabarcoding for a relative quantitative analysis. These results move towards the application of HTS analysis for studying the composition of herbal teas for medicinal plants’ traceability and quality control.

In the context of novel foods, a category for which the market demand is increasing worldwide, the consumption of edible insects and related insect-based products is expected to grow in the next years. Insects represent an important source of energy for the human diet but there is a lack of scientific knowledge about their processing to ensure safe food items to the consumer. In this study we adopted a combined DNA-based approach to verify the identity of the declared species in five categories of commercial insect-based products (mt COI DNA barcoding) and to characterize plant declared ingredients or contaminants (nu ITS2 DNA metabarcoding) with particular attention to putative elements of allergenic concern belonging, for example to the insect rearing substrate. Moreover, the same approach has been used to assess its sensitivity to cases of contamination and counterfeits to insect flour with low cost (and potentially allergenic) vegetable flours like wheat and soybean. Results show the success of insect DNA barcoding authentication even for highly processed products. Furthermore, the DNA metabarcoding analysis revealed a high efficacy as a screening method to identify both plant ingredients and vegetal traces belonging to insect farming or possible adulteration events, also acting as an early warning strategy for the occurrence of allergens of human concern. This approach could support the development of new risk assessment procedures for novel foods by regulatory authorities to ensure their quality, safety, and acceptance which will become more required in order to face the challenge of feeding the world population in the next decades.

The spread of food allergens is a topic of global importance due to its impact on public health. National and International regulations ask food producers and manufacturers to declare product compositions on the label, especially in case of processed raw materials. Wheat flour (Triticum aestivum) can be contaminated by a wide range of species belonging to the Brassicaceae in the field or during grain harvests, storage, and processing. Among them, mustards (Brassica nigra, Brassica juncea and Sinapis alba) are well known allergenic species. Often, food quality laboratories adopt an ELISA approach to detect the presence of mustard species. However, this approach shows cross-reactivity with other non-allergenic species such as Brassica napus (rapeseed). In the last few years, DNA barcoding was proposed as a valid identification method, and it is now commonly used in the authentication of food products. This study aims to set up an easy and rapid DNA-based tool to detect mustard allergenic species. DNA barcoding (matK and ITS2) and chromosome markers (A6, B, C1 genome regions) were selected, and specific primers were validated on incurred reference food matrices. The developed test was proven to be able to distinguish mustard from rapeseed and wheat, overcoming cross-reactivity with Brassica napus.

The global market of food supplements is growing, along with consumers demand for high-quality herbal products. Nevertheless, substitution fraud, and adulteration cases remain a common safety problem of global concern. In the last years, the DNA barcoding approach has been proposed as a valid identification method and it is now commonly used in the authentication of herbal and food pro-ducts. The objective of this study was to evaluate whether DNA barcoding can be applied to trace the plant species from the starting raw material to the finished commercial products. We selected a panel of 28 phytoextracts obtained through three different extraction methods (i.e., maceration, percolation and sonication) with different solvents (i.e., ethanol, deionized water and glycerol). Furthermore, we chose six plant species for which we collected and analysed all the intermediates of the industrial production. We sequenced and analyzed the sequence variability at DNA barcoding (psbA-trnH, ITS) and minibarcoding (rbcL 1-B) marker regions. Phytoextracts obtained through hydroalcoholic treatment, with the lower per-centage of ethanol (<40%), and aqueous processing, at the lowest temperature, had major rate of sequencing and identification success. This study proves that DNA barcoding is a useful tool for Medicinal and Aromatic Plants (MAPs) traceability, which would provide consumers with safe and high-quality herbal products.

In this contribution, new data concerning the distribution of vascular flora alien to Italy are presented. It includes new records, confirmations, exclusions, and status changes for Italy or for Italian administrative regions of taxa in the genera Bunias, Calocedrus, Calycanthus, Celosia, Clerodendrum, Convolvulus, Crassula, Cyclamen, Datura, Dicliptera, Eragrostis, Erigeron, Gamochaeta, Gazania, Impatiens, Kolkwitzia, Leucaena, Ludwigia, Medicago, Muscari, Nigella, Oenothera, Opuntia, Paulownia, Petroselinum, Phyllostachys, Physalis, Pseudosasa, Quercus, Reynoutria, Roldana, Saccharum, Sedum, Semiarundinaria, Senecio, Sisyrinchium, Solanum, Sporobolus, Tulipa, Vachellia, Verbena, and Youngia. Nomenclatural and distribution updates published elsewhere are provided as Suppl. material 1.

La sicurezza e la qualità degli alimenti dipendono dalle caratteristiche delle materie prime e dagli approcci chimici, fisici e biotecnologici adottati durante i processi di produzione. Poiché un gran numero di microrganismi sono coinvolti, la maggior parte dei prodotti dovrebbe essere considerata come matrici complesse in cui ogni componente microbico ha un ruolo preciso e si evolve in risposta ai cambiamenti dell'intero sistema. Inoltre, la comprensione delle dinamiche microbiche coinvolte nella produzione alimentare è utile per migliorare i processi industriali e il prodotto stesso. L'analisi del microbiota alimentare è fondamentale per migliorare i processi di biotrasformazione, come la vinificazione. Da tempo l’industria del vino ha coltivato selettivamente cultivar di vite che mostrano tratti diversi (dimensioni, colore, sapore dell'uva) e piccole variazioni nella composizione del suolo, nell'irrigazione e del clima sono da tempo associate a cambiamenti in questi tratti. La flora microbica che coesiste con la pianta può essere uno dei fattori chiave che influenzano queste peculiarità. Le tecnologie High Throughput Sequencing (HTS) sono al giorno d'oggi uno strumento emergente e ampiamente adottato per la caratterizzazione microbica, anche nelle matrici alimentari. Per chiarire il contributo del microbioma della vite durante le fasi di fermentazione, ho utilizzato un approccio basato sull'HTS per identificare le comunità di batteri e funghi associate a bacche e mosti della cultivar Cannonau, provenienti da quattro vigneti di diverse regioni della Sardegna. Il Cannonau è la più importante cultivar di vite della Sardegna (Italia), dove la maggior parte dei vigneti non subisce senza trattamenti fitochimici. Le analisi bioinformatiche hanno suggerito che i microbiomi delle bacche raccolte condividevano una composizione batterica caratterizzata da Enterobacteriales, Pseudomonadales, Bacillales e Rhodospirillales. Tuttavia, ogni area sembra arricchire i micr, Food safety and quality depend on raw material characteristics and on the chemical, physical and biotechnological approaches adopted during manufacturing and transformation processes. Since a huge number of microorganisms are involved in food production, most products should be considered as complex matrices where any microbial component has a precise role and evolves in response to changes in physical and chemical features of the whole system. So, understanding the dynamics of microbial community involved in a food supply chain is useful to reduce food spoilage outbreaks, enhance industrial processes and extend product’s shelf-life. The analysis of food microbiota is also pivotal to improve biotransformation processes, like winemaking. From time immemorial, the wine industry has been selectively growing grapevine cultivars showing different traits (e.g., grape size, color and flavor), and small variations in soil composition, irrigation and climate, have long been associated with shifts in these traits. The microbial flora coexisting with the plant may be one of the key factors influencing these traits. Despite long-time difficulties in analyzing single bacterial strains, the High Throughput Sequencing technologies (HTS) are nowadays an emerging and widely adopted tool for microbial characterization, even in food matrices. To clarify the contribution of the microbiome of grape during wine fermentation steps, I used an HTS-based approach to identify bacteria and fungi communities associated to berries and musts of Cannonau cultivar from four vineyards belonging to different regions in Sardinia. Cannonau is the most important grapevine cultivar of Sardinia (Italy), where most vineyards are cultivated without phytochemical treatments. Bioinformatics analyses suggested that microbiome colonizing berries collected at the four different localities shared a core composition characterized by Enterobacteriales, Pseudomonadales, Bacillales, and Rhodospirillales. However, any

Recent studies have highlighted the role of the grapevine microbiome in addressing a wide panel of features, ranging from the signature of field origin to wine quality. Although the influence of cultivar and vineyard environmental conditions in shaping the grape microbiome have already been ascertained, several aspects related to this topic, deserve to be further investigated. In this study, we selected three international diffused grapevine cultivars (Cabernet Sauvignon, Syrah, and Sauvignon Blanc) at three germplasm collections characterized by different climatic conditions [Northern Italy (NI), Italian Alps (AI), and Northern Spain (NS)]. The soil and grape microbiome was characterized by 16s rRNA High Throughput Sequencing (HTS), and the obtained results showed that all grape samples shared some bacterial taxa, regardless of sampling locality (e.g., Bacillus, Methylobacterium, Sphingomonas, and other genera belonging to Alphaproteobacteria, Gammaproteobacteria, and Actinobacteria). However, some Operational Taxonomic Units (OTUs) could act as geographical signatures and in some cases as cultivar fingerprint. Concerning the origin of the grape microbiome, our study confirms that vineyard soil represents a primary reservoir for grape associated bacteria with almost 60% of genera shared between the soil and grape. At each locality, grapevine cultivars shared a core of bacterial genera belonging to the vineyard soil, as well as from other local biodiversity elements such as arthropods inhabiting or foraging in the vineyard. Finally, a machine learning analysis showed that it was possible to predict the geographical origin and cultivar of grape starting from its microbiome composition with a high accuracy (9 cases out of 12 tested samples). Overall, these findings open new perspectives for the development of more comprehensive and integrated research activities to test which environmental variables have an effective role in shaping the microbiome composition and dyna

Grape berries harbor a wide range of microbes originating from the vineyard environment, many of which are recognized for their role in the must fermentation process shaping wine quality. To better clarify the contribution of the microbiome of grape fruits during wine fermentation, we used high-throughput sequencing to identify bacterial and fungi communities associated with berries and musts of Cannonau. This is the most important cultivar-wine of Sardinia (Italy) where most vineyards are cultivated without phytochemical treatments. Results suggested that microbiomes of berries collected at four different localities share a core composition characterized by Enterobacteriales, Pseudomonadales, Bacillales, and Rhodospirillales. However, any area seems to enrich berries microbiome with peculiar microbial traits. For example, berries belonging to the biodynamic vineyards of Mamoiada were rich in Bacillales typical of manure (i.e. Lysinibacillus, Bacillus, and Sporosarcina), whereas in the Santadi locality, berries showed soil bacteria such as Pasteurellales and Bacteroidales as well as Rhodospirillales and Lactobacillales which are commonly involved in wine fermentation. In the case of fungi, the most abundant taxa were Dothioraceae, Pleosporaceae, and Saccharomycodaceae, and although the proportion of these families varied among localities, they occurred ubiquitously in all vineyards. During vinification processes performed at the same wine cellar under controlled conditions and without using any yeast starter, more than 50% of bacteria groups of berries reached musts, and each locality had its own private bacteria signature, even if Saccharomyces cerevisiae represented the most abundant fungal species. This work suggests that natural berries microbiome could be influenced by pedoclimatic and anthropologic conditions (e.g., farming management), and the fruits’ microorganisms persist during the fermentation process. For these reasons, a reliable wine genotyping shoul

Food trade globalization and the growing demand for selected food varieties have led to the intensification of adulteration cases, especially in the form of species substitution and mixing with cheaper taxa. This phenomenon has huge economic impact and sometimes even public health implications. DNA barcoding represents a well-proven molecular approach to assess the authenticity of food items, although its use is hampered by analytical constraints and timeframes that are often prohibitive for the food market. To address such issues, we have introduced a new technology, named NanoTracer, that allows for rapid and naked-eye molecular traceability of any food and requires limited instrumentation and cost-effective reagents. Moreover, unlike sequencing, this method can be used to identify not only the substitution of a fine ingredient, but also its dilution with cheaper ones.

Purpose: The aim of this study was to evaluate in the vagina of 60 pre-menopausal women the detection of orally administered multispecies probiotic formulations showing anti-microbial properties in test in vitro. Methods: A randomized, double-blind, three-arm parallel pilot study was carried out on 60 pre-menopausal women. Subjects were randomly divided in three groups (F_1, F_2, F_3). Each group received a daily oral administration of probiotic mixtures (for 14 days and at the day 21, 7 days after the wash-out) containing: Lactobacillus acidophilus and Lactobacillus reuteri (F_1), or Lactobacillus plantarum, Lactobacillus rhamnosus and Bifidobacterium animalis subsp. lactis (F_2), or placebo (F_3), respectively. Vaginal swabs were collected at four experimental times, at t0 and at t7, t14 and t21 days, and analyzed by qPCR. At the same time, the anti-microbial activity of the probiotic formulations was verified by assays in vitro against microorganisms as Escherichia coli and Candida albicans. Results: L. acidophilus and L. reuteri as well as L. plantarum, L. rhamnosus and B. lactis were significantly increased on 7 days in the groups administered with F_1 and F_2, respectively, compared to group F_3. A similar significant trend was observed on 21 days, 7 days after the wash-out. F_1 and F_2 showed coherent anti-microbial properties. Conclusion: Both probiotic formulations F_1 and F_2, chosen because of their anti-microbial activity against pathogens responsible for vaginal dysbiosis and infections, led to vaginal detection and enhancement of the amount of species of formulates when orally administered. This work provides the basis for further clinical investigations of the F_1 and F_2 capacity to prevent or treat uro-genital infections.

The progressive loss of knowledge about the use of plants in traditional medicine practices is an issue of major concern with social, economic, and health implications for local communities, especially in developing countries. The Peruvian Amazon is one of the principal scenarios where this phenomenon is occurring at a growing rate. We investigated the case of Piri-Piri (Cyperus articulatus L.), a species of sedge used by native populations of Peru as an antiemetic, an anti-diarrheal, a sedative, an anti-venom, and to treat other symptoms or to conduct ritual practices. In this study, we collected 19 samples of Piri-Piri from different natives' familiar nuclei in four Amazonian villages, and we used the molecular tool DNA barcoding to verify the corrected species identification. The identification performances of four barcode loci (rbcL, matK, trnH-psbA and ITS) were evaluated. RbcL and matK were not able to univocally identify the Piri-Piri samples; however, they indicated that one sample did not match with the expected C. articulatus (local misidentification). This finding was further confirmed at both trnH-psbA and ITS regions. These two loci designated the remaining 18 samples to the correct species, C. articulatus. Based on our results, we propose DNA barcoding as a reliable tool for preserving the genetic identity and the exploitation of Piri-Piri by the local traditional medicine. This universal and fast molecular characterization method should be included in Internet-based Automated Identification Technology systems to enhance the spread of traditional medicine and prevent the erosion of ancient traditional knowledge.

Plant exposures are among the most frequently reported cases to poison control centres worldwide. This is a growing condition due to recent societal trends oriented towards the consumption of wild plants as food, cosmetics, or medicine. At least three general causes of plant poisoning can be identified: plant misidentification, introduction of new plant-based supplements and medicines with no controls about their safety, and the lack of regulation for the trading of herbal and phytochemical products. Moreover, an efficient screening for the occurrence of plants poisonous to humans is also desirable at the different stages of the food supply chain: from the raw material to the final transformed product. A rapid diagnosis of intoxication cases is necessary in order to provide the most reliable treatment. However, a precise taxonomic characterization of the ingested species is often challenging. In this review, we provide an overview of the emerging DNA-based tools and technologies to address the issue of poisonous plant identification. Specifically, classic DNA barcoding and its applications using High Resolution Melting (Bar-HRM) ensure high universality and rapid response respectively, whereas High Throughput Sequencing techniques (HTS) provide a complete characterization of plant residues in complex matrices. The pros and cons of each approach have been evaluated with the final aim of proposing a general user’s guide to molecular identification directed to different stakeholder categories interested in the diagnostics of poisonous plants.

Background and Aim. The efficacy of supplementation treatment with two multispecies probiotic formulates on subjects diagnosed with IBS-C and the assessment of their gut microbiota were investigated. Methods. A randomized, double-blind, three-arm parallel group trial was carried out on 150 IBS-C subjects divided into three groups (F-1, F-2, and F-3). Each group received a daily oral administration of probiotic mixtures (for 60 days) F-1 or F-2 or placebo F-3, respectively. Fecal microbiological analyses were performed by species-specific qPCR to assess the different amount of probiotics. Results. The percentage of responders for each symptom was higher in the probiotic groups when compared to placebo group during the treatment period (t 60) and was maintained quite similar during the follow-up period (t 90). Fecal analysis demonstrated that probiotics of the formulations increased during the times of treatment only in fecal DNA from subjects treated with F-1 and F-2 and not with F-3, and the same level was maintained during the follow-up period. Conclusions. Multispecies probiotic supplementations are effective in IBS-C subjects and induce a different assessment in the composition of intestinal microbiota. This clinical study is registered with the clinical study registration number ISRCTN15032219.

Vitis vinifera L. is largely diffused in Sardinia, where it occurs as both a spontaneous wild plant (V. vinifera subsp. silvestris) and a crop (V. vinifera subsp. vinifera). The huge number of grapevine cultivars is the result of the complex history of Sardinia. The island was part of the Spanish Empire from 1479 to 1714 when commercial and cultural exchanges between the two regions caused modifications of its agricultural products. Noticeably, in the case of viticulture, Spanish and Sardinian grapevine varieties often have similar-sounding names; for example, ‘Bobal’ and ‘Bovale’, ‘Cariñena’ and ‘Carignano’, and ‘Garnacha’, ‘Granaccia’ and ‘Granazza’. We may assume that, due to politic and economic relationships, exchanges of grape varieties could have been occurred thus improving the genetic composition of the local germplasm. At the same time, certain Sardinian cultivars, some of which derived from the domestication and cultivation of wild local grapevines, could have been introduced in Spain. The first aim of our work was to evaluate the genetic structure of grapevine germplasm of Sardinia and its relationships with Spanish cultivars. Simple Sequence Repeat (SSR) markers were analyzed to characterize each accession and to study the relationships between Sardinian and Spanish cultivars. Results suggest the occurrence of synonymies such as ‘Cannonau’ and several Spanish accessions of ‘Garnacha Tinta’. If genetic origin of cultivars is an element useful for assessing wine traceability, wine quality relies on different factors and it is largely influenced by the environmental conditions of the production area. The second goal of this study was to develop an multidisciplinary approach for the characterization of 'Cannonau' wine produced in different Sardinian localities. We conducted a high-throughput sequencing analysis of different samples to understand the relationship between wine identity and the microbiome, specifically concerning bacteria and fungi. Since mic

Frugivores are critical components of restoration programs because they are seed dispersers. Thus, knowledge about bird-plant trophic relationships is essential in the evaluation of the efficacy of restoration processes. Traditionally, the diet of frugivores is characterized by microscopically identifying plant residues in droppings, which is time-consuming, requires botanical knowledge, and cannot be used for fragments lacking detectable morphological characteristics (e.g., fragmented seeds and skins). We examined whether DNA barcoding can be used as a universal tool to rapidly characterize the diet of a frugivorous bird, Eurasian blackcap (Sylvia atricapilla). We used the DNA barcoding results to assess restoration efforts and monitor the diversity of potentially dispersed plants in a protected area in northern Italy. We collected 642 Eurasian Blackcap droppings at the restored site during the autumn migration over 3 years. Intact seeds and fragmented plant material were analyzed at 2 plastidial barcode loci (rbcL and trnH-psbA), and the resulting plant identifications were validated by comparison with a reference molecular data set of local flora. At least 17 plant species, including 7 of the 11 newly transplanted taxa, were found. Our results demonstrate the potential for DNA barcoding to be used to monitor the effectiveness of restoration plantings and to obtain information about fruit consumption and dispersal of invasive or unexpected plant species. Such an approach provides valuable information that could be used to study local plant biodiversity and to survey its evolution over time.

In this study, Hypnum cupressiforme moss bags were used to examine the atmospheric deposition of trace elements in the oil refinery region of Sardinia (Italy) compared with surrounding natural zones. The concentrations of 13 elements [arsenic (As), calcium (Ca), cadmium (Cd), chromium (Cr), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), sodium (Na), nickel (Ni), lead (Pb), vanadium (V), and zinc (Zn)] were determined using inductively coupled plasma optical emission spectrometry. A significant accumulation of pollutants was detected using active biomonitoring with moss bags compared with a control site. The most relevant contaminants for all of the tested sites were Cr, Cu, Ni, and Zn. Moreover, the accumulation of Cr and Zn in the refinery industrial areas, IA1 and IA2, was more than five times greater than that detected at the control site. Levels of Cd, Mg, and Pb were also higher at all of the monitored sites compared with the control site. Both genomic and proteomic methods were used to study the response of H. cupressiforme to air pollution. No DNA damage or mutations were detected using the amplified fragment length polymorphisms (AFLP) method. At the protein level, 15 gel spots exhibited differential expression profiles between the moss samples collected at the IA1 site and the control site. Furthermore, among the 14 spots that showed a decrease in protein expression, nine were associated with ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and proteins of the light-harvesting complexes of photosystem (PS) II, three were associated with protein synthesis, and three were stress-related proteins. Thus, some of these proteins may represent good moss biosensors which could be used as pre-alert markers of environmental pollution.

In the last decades, food science has greatly developed, turning from the consideration of food as mere source of energy to a growing awareness on its importance for health and particularly in reducing the risk of diseases. Such vision led to an increasing attention towards the origin and quality of raw materials as well as their derived food products. The continuous advance in molecular biology allowed setting up efficient and universal omics tools to unequivocally identify the origin of food items and their traceability. In this review, we considered the application of a genomics approach known as DNA barcoding in characterizing the composition of foodstuffs and its traceability along the food supply chain. Moreover, metabolomics analytical strategies based on Nuclear Magnetic Resonance (NMR) and Mass Spectroscopy (MS) were discussed as they also work well in evaluating food quality. The combination of both approaches allows us to define a sort of molecular labelling of food that is easily understandable by the operators involved in the food sector: producers, distributors, and consumers. Current technologies based on digital information systems such as web platforms and smartphone apps can facilitate the adoption of such molecular labelling.

Probiotic ingestion is recommended as a preventive approach to maintain the balance of the intestinal microbiota and to enhance the human well-being. During the whole life of each individual, the gut microbiota composition could be altered by lifestyle, diet, antibiotic therapies and other stress conditions, which may lead to acute and chronic disorders. Hence, probiotics can be administered for the prevention or treatment of some disorders, including lactose malabsorption, acute diarrhoea, irritable bowel syndrome, necrotizing enterocolitis and mild forms of inflammatory bowel disease. The probiotic-mediated effect is an important issue that needs to be addressed in relation to strain-specific probiotic properties. In this work, the probiotic properties of new Lactobacillus and Bifidobacterium strains were screened, and their effects in vitro were evaluated. They were screened for probiotic properties by determining their tolerance to low pH and to bile salts, antibiotic sensitivity, antimicrobial activity and vitamin B8, B9 and B12 production, and by considering their ability to increase the antioxidant potential and to modulate the inflammatory status of systemic-miming cell lines in vitro. Three out of the examined strains presenting the most performant probiotic properties, as Lactobacillus plantarum PBS067, Lactobacillus rhamnosus PBS070 and Bifidobacterium animalis subsp. lactis PBSO75, were evaluated for their effects also on human intestinal HT-29 cell line. The obtained results support the possibility to move to another level of study, that is, the oral administration of these probiotical strains to patients with acute and chronic gut disorders, by in vivo experiments.

The study of diversity in biological communities is an intriguing field. Huge amount of data are nowadays available (provided by the innovative DNA sequencing techniques), and management, analysis and display of results are not trivial. Here, we propose for the first time the use of phylogenetic entropy as a measure of bacterial diversity in studies of microbial community structure. We then compared our new method (i.e. the web tool phyloh) for partitioning phylogenetic diversity with the traditional approach in diversity analyses of bacteria communities. We tested phyloh to characterize microbiome in the honeybee (Apis mellifera, Insecta: Hymenoptera) and its parasitic mite varroa (Varroa destructor, Arachnida: Parasitiformes). The rationale is that the comparative analysis of honeybee and varroa microbiomes could open new perspectives concerning the role of the parasites on honeybee colonies health. Our results showed a dramatic change of the honeybee microbiome when varroa occurs, suggesting that this parasite is able to influence host microbiome. Among the different approaches used, only the entropy method, in conjunction with phylogenetic constraint as implemented in phyloh, was able to discriminate varroa microbiome from that of parasitized honeybees. In conclusion, we foresee that the use of phylogenetic entropy could become a new standard in the analyses of community structure, in particular to prove the contribution of each biological entity to the overall diversity.

Food safety and quality depend on raw material characteristics and on the chemical, physical and biotechnological processes adopted during food transformation. Since a huge number of microorganisms are involved in food production, foodstuffs should be considered as complex matrices where any microbial component has a precise role and evolves in response to physical and chemical composition of food. Moreover, knowing the dynamics of microbial community involved in a food supply chain it is useful to reduce food spoilage, enhance the industrial processes and extend product shelf-life. In a more comprehensive vision, a precise understanding of the metabolic activity of microorganisms can be used to drive biotransformation steps towards the improvement of quality and nutritional value of food. High Throughput Sequencing (HTS) technologies are nowadays an emerging and widely adopted tool for microbial characterization of food matrices. Differently from traditional culture-dependent approaches, HTS allows the analysis of genomic regions of the whole biotic panel inhabiting and constituting food ecosystems. Our intent is to provide an up-to-date review of the principal fields of application of HTS in food studies. In particular, we devoted major attention to the analysis of food microbiota and to the applied implications deriving from its characterization in the principal food categories to improve biotransformation processes.

In this study, the genetic constitution of wild grape populations distributed in the Zagros Mountains in the north-west part of Iran was investigated to assess their risk of extinction and to identify the most suitable conservation strategy. A total of 65 wild grapevine samples from five different populations were analysed using 23 SSR loci. Consistent allelic richness was detected in all the analysed populations (from 4.01 to 5.35). The observed heterozygosity and expected heterozygosity were high with mean values of 0.67 and 0.69, respectively. A modest heterozygosity deficiency was detected only in the Piranshahr and Sardasht-Shalmash populations. The neighbour-joining dendrogram clustered the accessions into three groups corresponding to the geographical distribution of the populations. Molecular data combined with field evaluation revealed that an ex situ conservation approach is the most suitable strategy for preserving the genetic richness of Iranian wild grape populations with consistent rare alleles (about 20%). We propose in situ preservation only in the case of the Sardasht-Ghasmarash population to ensure that evolutionary dynamic forces continue to influence plant adaptation and survival in response to environmental alterations.

This outlook paper addresses the problem of the traceability of minor crops. These kinds of cultivations consist in a large number of plants locally distributed with a modest production in terms of cultivated acreage and quantity of final product. Because of globalization, the diffusion of minor crops is increasing due to their benefit for human health or their use as food supplements. Such a phenomenon implies a major risk for species substitution or uncontrolled admixture of manufactured plant products with severe consequences for the health of consumers. The need for a reliable identification system is therefore essential to evaluate the quality and provenance of minor agricultural products. DNA-based techniques can help in achieving this mission. In particular, the DNA barcoding approach has gained a role of primary importance thanks to its universality and versatility. Here, we present the advantages in the use of DNA barcoding for the characterization and traceability of minor crops based on our previous or ongoing studies at the ZooPlantLab (Milan, Italy). We also discuss how DNA barcoding may potentially be transferred from the laboratory to the food supply chain, from field to table.