DNA barcoding to trace Medicinal and Aromatic Plants from the field to the food supplement

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 products. 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 percentage 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

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10&nbsp;years; 78.2% included were male with a median age of 37&nbsp;years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

DNA-Based Herbal Teas’ Authentication: An ITS2 and psbA-trnH Multi-Marker DNA Metabarcoding Approach

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

Natural Deep Eutectic Solvents (NADESs) Combined with Sustainable Extraction Techniques: A Review of the Green Chemistry Approach in Food Analysis

Usual extraction processes for analyzing foods, supplements, and nutraceutical products involve massive amounts of organic solvents contributing to a negative impact on the environment and human health. In recent years, a new class of green solvents called natural deep eutectic solvents (NADES) have been considered a valid alternative to conventional solvents. Compared with conventional organic solvents, NADES have attracted considerable attention since they are sustainable, biodegradable, and non-toxic but also are easy to prepare, and have low production costs. Here we summarize the major aspects of NADEs such as the classification, preparation method physicochemical properties, and toxicity. Moreover, we provide an overview of novel extraction techniques using NADES as potential extractants of bioactive compounds from foods and food by-products, and application of NADEs in food analysis. This review aims to be useful for the further development of NAES and for broadening the knowledge of these new green solvents in order to increase their use for the extraction of bioactive compounds and in food analysis

THE PROBLEM OF MISIDENTIFICATION BETWEEN A WILD EDIBLE PLANT AND A POISONOUS ONE: THE CASE OF BORAGE

At present, in many European countries people is looking for wild edible plants to experience new tastes and flavors, by following the new trend of being green and environmentally friendly (1). Borago officinalis L. (borage) is an annual herb originating in the Mediterranean region but naturalized and widely cultivated throughout most of Europe, traditionally used for culinary and medicinal purposes. Its leaves are mixed in salads and used as a vegetable in different European countries, such as Germany, Spain, Greece, and Italy (2), especially in Liguria as a stuff of traditional ravioli and pansoti (3). However, young borage leaves can be easily confused by inexpert pickers with those of other plants, including poisonous ones, such as Mandragora autumnalis (mandrake), common in Southern Italy and Sicily, or Digitalis purpurea (foxglove), in Northern Italy. Patients who turn to Italian Poison Control Centers or Hospital Emergency Rooms after accidental ingestion of these plants, show anticholinergic symptoms due to unintentional ingestion of the leaves of mandrake (most commonly) or foxglove (less frequently). In the period 1995-2007, 50 cases of intoxication by accidental ingestion of mandrake and 6 cases after ingestion of foxglove have been reported in Italy (1). In the present work we show the pharmacognostic characterization of young leaves from B. officinalis (Boraginaceae), M. autumnalis (Solanaceae), and D. purpurea (Scrophulariaceae). Micromorphological, phytochemical and molecular identification techniques were used. Fresh leaf samples were analyzed by optical and scanning electron microscopy, highlighting main anatomical and histological features, such as stomata and trichome types and distribution (Fig 1, A-C). DNA barcoding sequences (using a region of the plastidial RbcL as DNA marker), analyzed using a bioinformatics tool (MEGA v. 7.0), allowed to determine the intra- and inter-genetic variability (K2P distance) among the three taxa. No genetic variation was detected within species, while consistent genetic distances were observed among species: borage vs foxglove K2P = 7.46%, foxglove vs. mandrake K2P = 7.46% and borage vs mandrake K2P = 7.46%. GC-MS analysis of fresh leaves pentane/hexane extract (5:1, v/v) revealed a typical chemical fingerprint of each plant analyzed and a particularly interesting difference between poisonous and nonpoisonous plants. In fact, M. autumnalis and D. purpurea leaf extract contain vitamin E, while in B. officinalis this metabolite is absent. Therefore, the authors conclude that, in this case, the presence of this metabolite is discriminant among poisonous and edible plants and could be used as a phytochemical marker, while among the poisonous ones becomes useful analyze the discriminant wax alkanes to differentiate them. This study provides a multi-disciplinary approach to the problem of misidentification between wild edible plants and poisonous species. The reported protocols provide fast and reliable determination of species causing poisoning, allowing a quick management of poisoned patients

Food Tracking Perspective: DNA Metabarcoding to Identify Plant Composition in Complex and Processed Food Products

One of the main goals of the quality control evaluation is to identify contaminants in raw material, or contamination after a food is processed and before it is placed on the market. During the treatment processes, contamination, both accidental and economically motivated, can generate incongruence between declared and real composition. In our study, we evaluated if DNA metabarcoding is a suitable tool for unveiling the composition of processed food, when it contains small trace amounts. We tested this method on different types of commercial plant products by using tnrL marker and we applied amplicon-based high-throughput sequencing techniques to identify plant components in different food products. Our results showed that DNA metabarcoding can be an effective approach for food traceability in different type of processed food. Indeed, the vast majority of our samples, we identified the species composition as the labels reported. Although some critical issues still exist, mostly deriving from the starting composition (i.e., variable complexity in taxa composition) of the sample itself and the different processing level (i.e., high or low DNA degradation), our data confirmed the potential of the DNA metabarcoding approach also in quantitative analyses for food composition quality control

Check Your Shopping Cart: DNA Barcoding and Mini-Barcoding for Food Authentication

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