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In a recent paper, "Environmental DNA: What's behind the term? Clarifying the terminology and recommendations for its future use in biomonitoring," Pawlowski et al. argue that the term eDNA should be used to refer to the pool of DNA isolated from environmental samples, as opposed to only extra-organismal DNA from macro-organisms. We agree with this view. However, we are concerned that their proposed two-level terminology specifying sampling environment and targeted taxa is overly simplistic and might hinder rather than improve clear communication about environmental DNA and its use in biomonitoring. This terminology is based on categories that are often difficult to assign and uninformative, and it overlooks a fundamental distinction within eDNA: the type of DNA (organismal or extra-organismal) from which ecological interpretations are derived., Molecular Ecology, 30 (19), ISSN:0962-1083, ISSN:1365-294X

The promotion of responsible and sustainable trade in biological resources is widely proposed as one solution to mitigate currently high levels of global biodiversity loss. Various molecular identification methods have been proposed as appropriate tools for monitoring global supply chains of commercialized animals and plants. We demonstrate the efficacy of target capture genomic barcoding in identifying and establishing the geographic origin of samples traded as Anacyclus pyrethrum, a medicinal plant assessed as globally vulnerable in the IUCN Red List of Threatened Species. Samples collected from national and international supply chains were identified through target capture sequencing of 443 low-copy nuclear makers and compared to results derived from genome skimming of plastome, standard plastid barcoding regions and ITS. Both target capture and genome skimming provided approximately 3.4 million reads per sample, but target capture largely outperformed standard plant DNA barcodes and entire plastid genome sequences. Despite the difficulty of distinguishing among closely related species and infraspecific taxa of Anacyclus using conventional taxonomic methods, we succeeded in identifying 89 of 110 analysed samples to subspecies level without ambiguity through target capture. Of the remaining samples, we determined that eleven contained plant material from other genera and families and ten were unidentifiable regardless of the method used. Furthermore, we were able to discern the geographical origin of Anacyclus samples collected in Moroccan, Indian and Sri Lankan markets, differentiating between plant materials originally harvested from diverse populations in Algeria and Morocco. With a recent drop in the cost of analysing samples, target capture offers the potential to routinely identify commercialized plant species and determine their geographic origin. It promises to play an important role in monitoring and regulation of plant species in trade, supporting biodiversity conservation efforts, and in ensuring that plant products are unadulterated, contributing to consumer protection.

International audience; Global vegetation and land-surface models embody interdisciplinary scientific understanding of the behaviour of plants and ecosystems, and are indispensable to project the impacts of environmental change on vegetation and the interactions between vegetation and climate. However, systematic errors and persistently large differences among carbon and water cycle projections by different models highlight the limitations of current process formulations. In this review, focusing on core plant functions in the terrestrial carbon and water cycles, we show how unifying hypotheses derived from eco-evolutionary optimality (EEO) principles can provide novel, parameter-sparse representations of plant and vegetation processes. We present case studiesthat demonstrate how EEO generate parsimonious representations of core, leaf-level processes that are individually testable and supported by evidence. EEO approaches to photosynthesis and primary production, dark respiration, and stomatal behaviour are ripe for implementation in global models. EEO approaches to other important traits, including the leaf economics spectrum and applications of EEO at the community level are active research areas. Independently tested modules emerging from EEO studies could profitably be integrated into modelling frameworks that account for the multiple time scales on which plants and plant communities adjust toenvironmental change.

Water is an essential resource for human life and the environment. The widespread use of chemicals in daily life has led to significant water quality concerns. During the production phase, the use phase as well as after use, (residues of) these chemicals can enter the environment and water systems. Furthermore, the use and production of chemicals are increasing rapidly, driven by mainly population growth, urbanisation and economic growth. Increased use leads to further emissions of chemicals to water, posing significant water quality concerns. Henceforth there is an urgent need to understand the linkage between society and production and consumption of chemicals to explore possible changes in water quality.Socio-economic scenario analysis is a useful tool to investigate the long-term consequences of future change and mitigation options. While scenarios have been broadly applied to understand air pollution, this not yet the case for chemical pollution to surface waters. In this work, we propose a general framework to develop scenarios for the future emissions of chemicals to water by using the Shared Socio-economic Pathways (SSP). The framework follows the basic elements of the scenario development process by defining the current system, describe the changes in emissions with scenario drivers and elaboration to the future. The framework is then tested on a set of selected ‘example’ chemicals that represent broader chemical groups of pharmaceuticals (Ibuprofen and Diclofenac), pesticides (Terbuthylazine) and industrial chemicals (Cadmium and Di-ethyl phthalate). Chemical emissions to water over the past years were used to understand their yearly trends and patterns over the European countries. Lastly, the emission scenarios for chemicals for 2050 were developed by using SSP drivers from the IMAGE Integrated assessment model as an input to the empirical emission models. The three SSP scenarios: SSP1 ("Sustainability"), SSP2 ("Middle of the Road") and SSP3 ("Regional Rivalry") focusing on Europe were included. Additionally, the developed scenarios also describe mitigation efforts.The results of emission scenarios displayed an increase in emissions up to 2050 for the exemplary chemicals in Western Europe for all three scenarios SSP1, SSP2 and SSP3. While the emissions of chemicals linearly decreased in Eastern Europe for the same period. SSP3 showed the highest emissions in 2050 except for cadmium emissions from wastewater treatment plants. The results showed that the framework helps in understanding the possible influence of socio-economic changes on use and emissions of chemicals which can be a part of future risk assessments. While the framework can be extended similarly to other pharmaceuticals and pesticides, it requires a detailed understanding of complex emission sources for industrial chemicals.

Monitoring of airborne pollen concentrations provides an important source of information for the globally increasing number of hay fever patients. Airborne pollen are traditionally counted under the microscope, but with the latest developments in image recognition methods, automating this process has become feasible. A challenge that persists, however, is that many pollen grains cannot be distinguished beyond the genus or family level using a microscope. Here, we assess the use of a Convolutional Neural Network (CNN) to increase taxonomic accuracy for airborne pollen. As a case study we use the nettle family (Urticaceae), which contains two main genera (Urtica and Parietaria) common in European landscapes which pollen cannot be separated by trained specialists. While pollen from Urtica species have very low allergenic relevance, those from several species of Parietaria are severely allergenic. We collect pollen from both fresh as well as from herbarium specimens and use these to train the CNN model VGG16. The model shows that Urticaceae pollen can be distinguished with 98.3% accuracy. We then apply our model on Urticaceae pollen collected from aerobiological samples and show that the genera can be confidently distinguished, despite the more challenging input images that are often overlain by debris. Our method can also be applied to other pollen families in the future and will thus help to make allergenic pollen monitoring more specific.

Background Stomata analysis using microscope imagery provides important insight into plant physiology, health and the surrounding environmental conditions. Plant scientists are now able to conduct automated high-throughput analysis of stomata in microscope data, however, existing detection methods are sensitive to the appearance of stomata in the training images, thereby limiting general applicability. In addition, existing methods only generate bounding-boxes around detected stomata, which require users to implement additional image processing steps to study stomata morphology. In this paper, we develop a fully automated, robust stomata detection algorithm which can also identify individual stomata boundaries regardless of the plant species, sample collection method, imaging technique and magnification level. Results The proposed solution consists of three stages. First, the input image is pre-processed to remove any colour space biases occurring from different sample collection and imaging techniques. Then, a Mask R-CNN is applied to estimate individual stomata boundaries. The feature pyramid network embedded in the Mask R-CNN is utilised to identify stomata at different scales. Finally, a statistical filter is implemented at the Mask R-CNN output to reduce the number of false positive generated by the network. The algorithm was tested using 16 datasets from 12 sources, containing over 60,000 stomata. For the first time in this domain, the proposed solution was tested against 7 microscope datasets never seen by the algorithm to show the generalisability of the solution. Results indicated that the proposed approach can detect stomata with a precision, recall, and F-score of 95.10%, 83.34%, and 88.61%, respectively. A separate test conducted by comparing estimated stomata boundary values with manually measured data showed that the proposed method has an IoU score of 0.70; a 7% improvement over the bounding-box approach. Conclusions The proposed method shows robust performance across multiple microscope image datasets of different quality and scale. This generalised stomata detection algorithm allows plant scientists to conduct stomata analysis whilst eliminating the need to re-label and re-train for each new dataset. The open-source code shared with this project can be directly deployed in Google Colab or any other Tensorflow environment.

Ethnopharmalogical relevance In Africa, traditional medicine is important for local healthcare and plants used for these purposes are commonly traded. Identifying medicinal plants sold on markets is challenging, as leaves, barks and roots are often fragmented or powdered. Vernacular names are often homonymic, and identification of material lacking sufficient morphological characters is time-consuming, season-dependent and might lead to incorrect assessments of commercialised species diversity. Aim of the study In this study, we identified cases of vernacular heterogeneity of medicinal plants using a tiered approach of literature research, morphology and DNA barcoding. Material and methods A total of 870 single ingredient medicinal plant samples corresponding to 452 local names were purchased from herbal markets in Dar-es-Salaam and Tanga, Tanzania, and identified using conventional methods as well as DNA barcoding using rbcL, matK and nrITS. Results Using conventional methods, we could identify 70% of samples to at least family level, while 62% yielded a DNA barcode for at least one of the three markers. Combining conventional methods and DNA barcoding, 76% of the samples could be identified to species level, revealing a diversity of at least 175 species in 65 plant families. Analysis of the market samples revealed 80 cases of multilingualism and over- and under-differentiation. Afzelia quanzensis Welw., Zanthoxylum spp., Allophylus spp. and Albizia anthelmintica Brongn. were the most evident cases of multilingualism and over-differentiation, as they were traded under 8–12 vernacular names in up to five local languages. The most obvious case of under-differentiation was mwingajini (Swahili), which matched to eight scientific species in five different plant families. Conclusions Use of a tiered approach increases the identification success of medicinal plants sold in local market and corroborates findings that DNA barcoding can elucidate the identity of material that is unidentifiable based on morphology and literature as well as verify or disqualify these identifications. Results of this study can be used as a basis for quantitative market surveys of fragmented herbal medicine and to investigate conservation issues associated with this trade.

Herbal medicinal products gain increasing popularity. The growing demand for herbal medicine along with a lack of regulation render herbal products subject to intentional adulteration. The substitution of costly ingredients with unlabelled plant-based fillers of inferior quality has been widely reported. Such fraudulent practices erode consumer trust, but can also pose serious health risks. In this work, 71 herbal medicinal products were randomly purchased from Greek markets and analysed using ITS2 metabarcoding for species identification. The aim was to investigate possible adulterations and assess the efficacy of metabarcoding in plant-based product authentication. Of the 131 detected species in our analyses, 87 were not listed on the product labels. This indicates a high level of adulteration and/or contamination during processing and distribution. Two toxic species, Chelidonium majus and Nicotiana tabacum, were also detected as major ingredients of two herbal mixtures for medicinal purposes. Furthermore, the detection of wheat in eight samples raises concerns for people with gluten intolerance. This study stresses the need for stricter quality control of herbal products. In addition, to overcome the limitations of metabarcoding and augment the approach we used Bar-HRM for the first time as a verification tool. The combination of metabarcoding with species-specific Bar-HRM analysisenhances the reliability of the results.

Traditional crop varieties are an important source of genetic diversity for crop adaptation and modern breeding. Landraces of Asian (Oryza sativa) and African (Oryza glaberrima) rice have been well studied on the continents where they were domesticated. However, their history of cultivation in northern South America is poorly understood. Here we reveal the rice diversity that is maintained by Maroons, descendants of enslaved Africans who fled to the interior forests of the Guianas ca. 300 years ago. We interviewed subsistence farmers who practice shifting cultivation along the Maroni and Lawa rivers that form the natural border between French Guiana and Suriname, and used ethnobotanical and morphological methods to identify around 50 varieties, of which 15 were previously undocumented. The genetic origin of these varieties was explored using the Angiosperms-353 universal probe set. Despite the large distances between sites and relative inaccessibility of the area, phenotypic and genetic diversity did not display any geographic structure, which is consistent with knowledge of seed exchange among members of the same ethnolinguistic group. Although improved US cultivars were introduced in Maroon villages in the 1940s, these have not displaced the traditional landraces, which are cherished for their taste and nutritious qualities and for their importance in Maroon spiritual life. The unique agricultural and ritual practices of Maroons confirm their role as custodians of rice diversity, a role that is currently under threat from external pressures and encroaching globalization. We expect that the rice diversity uncovered in this study represents only a fraction of the total diversity in the Guianas and may constitute a large untapped resource that holds promise for future rice improvement. Further efforts to inventory and preserve these landraces will help to protect a precious cultural heritage and local food security.

Recent data shows increasing numbers of hay fever patients, with approximately 10-30% of the population affected worldwide (Pawankar et al. 2011). This increase is most likely caused by prolonged and intensified pollen seasons which in turn have been linked to increased CO2 concentrations (Ziska et al. 2003, D'Amato et al. 2007, Albertine et al. 2014). Apart from this, especially in cities, the so-called ‘heat island effect’ enables exotic plant species to establish themselves there. In the Netherlands alone, six new species settle in cities on a yearly basis and some of these are severely allergenic (Denters 2004). Pollen concentrations in the air are currently monitored using pollen samplers that collect pollen on sticky traps. These are checked manually under the microscope, a process that requires highly trained specialists. Moreover, microscopic pollen identification rarely allows discrimination of pollen types at species or even genus level even though the allergenicity may be very different. While there has been progress in automating the microscope using machine learning, automatic microscopes have not been able to systematically identify pollen to the species level. We designed an automated approach identify a predefined set of pollen on microscopic pollen samples. We use 2D light microscope images and a confocal fluorescence microscope for 3D images to create a reference dataset of highly similar pollen species to train automated image recognition software, and compare the results. The most accurate method will be used to apply to a pollen sample time series (1970-present) to find trends in allergenic pollen species over time. Here I present the first results of this research and the challenges to overcome.

Ayurveda is one of the oldest systems of medicine in the world, but the growing commercial interest in Ayurveda based products has increased the incentive for adulteration and substitution within this herbal market. Fraudulent practices such as the use of undeclared fillers and use of other species of inferior quality is driven both by the increased as well as insufficient supply capacity of especially wild plant species. Developing novel strategies to exhaustively assess and monitor both the quality of raw materials and final marketed herbal products is a challenge in herbal pharmacovigilance. Seventy-nine Ayurvedic herbal products sold as tablets, capsules, powders, and extracts were randomly purchased via e-commerce and pharmacies across Europe, and DNA metabarcoding was used to assess the ability of this method to authenticate these products. Our analysis reveals that only two out of 12 single ingredient products contained only one species as labeled, eight out of 27 multiple ingredient products contained none of the species listed on the label, and the remaining 19 products contained 1 to 5 of the species listed on the label along with many other species not specified on the label. The fidelity for single ingredient products was 67%, the overall ingredient fidelity for multi ingredient products was 21%, and for all products 24%. The low level of fidelity raises concerns about the reliability of the products, and detection of threatened species raises further concerns about illegal plant trade. The study highlights the necessity for quality control of the marketed herbal products and shows that DNA metabarcoding is an effective analytical approach to authenticate complex multi ingredient herbal products. However, effort needs to be done to standardize the protocols for DNA metabarcoding before this approach can be implemented as routine analytical approaches for plant identification, and approved for use in regulated procedures.

BioDATA (Biodiversity Data for Internationalisation in Higher Education) is an international project to develop and deliver biodiversity data training for undergraduate and postgraduate students from Armenia, Belarus, Tajikistan, and Ukraine. By training early career (student) biodiversity scholars, we aim at turning the current academic and education biodiversity landscape into a more open-data-friendly one. Professional practitioners (researchers, museum curators, and collection managers involved in data publishing) from each country were also invited to join the project as assistant teachers (mentors). The project is developed by the Research School in Biosystematics - ForBio and the Norwegian GBIF-node, both at the Natural History Museum of the University of Oslo in collaboration with the Secretariat of the Global Biodiversity Information Facility (GBIF) and partners from each of the target countries. The teaching material is based on the GBIF curriculum for data mobilization and all students will have the opportunity to gain the respective GBIF certification. All materials are made freely available for reuse and even in this very early phase of the project, we have already seen the first successful reuse of teaching materials among the project partners. The first BioDATA training event was organized in Minsk (Belarus) in February 2019 with the objective to train a minimum of four mentors from each target country. The mentor-trainees from this event will help us to teach the course to students in their home country together with teachers from the project team. BioDATA mentors will have the opportunity to gain GBIF certification as expert mentors which will open opportunities to contribute to future training events in the larger GBIF network. The BioDATA training events for the students will take place in Dushanbe (Tajikistan) in June 2019, in Minsk (Belarus) in November 2019, in Yerevan (Armenia) in April 2020, and in Kiev (Ukraine) in October 2020. Students from each country are invited to express their interest to participate by contacting their national project partner. We will close the project with a final symposium at the University of Oslo in March 2021. The project is funded by the Norwegian Agency for International Cooperation and Quality Enhancement in Higher Education (DIKU).

BioDATA is an international project on developing skills in biodiversity data management and data publishing. Between 2018 and 2021, undergraduate and postgraduate students from Armenia, Belarus, Tajikistan, and Ukraine, have an opportunity to take part in the intensive courses to become certified professionals in biodiversity data management. They will gain practical skills and obtain appropriate knowledge on: international data standards (Darwin Core); data cleaning software, data publishing software such as the Integrated Publishing Toolkit (IPT), and preparation of data papers. Working with databases, creating datasets, managing data for statistical analyses and publishing research papers are essential for the everyday tasks of a modern biologist. At the same time, these skills are rarely taught in higher education. Most of the contemporary professionals in biodiversity have to gain these skills independently, through colleagues, or through supervision. In addition, all the participants familiarize themselves with one of the important international research data infrastructures such as the Global Biodiversity Information Facility (GBIF). The project is coordinated by the University of Oslo (Norway) and supported by the Global Biodiversity Information Facility (GBIF). The project is funded by the Norwegian Agency for International Cooperation and Quality Enhancement in Higher Education (DIKU).

Background Phytopharmaceuticals are increasingly popular as alternative medicines, but poorly regulated in many countries. The manufacturers of these products should be subject to strict controls regarding each product's quality and constituents. Routine testing and identification of raw materials should be performed to ensure that the raw materials used in pharmaceutical products are suitable for their intended use. Hypothesis/purpose We have applied DNA Barcoding – High Resolution Melting (Bar-HRM), an emerging method for identifying of medicinal plant species based on DNA dissociation kinetics and DNA barcoding, for the authentication of medicinal plant species. Study design Commonly commercialized Thai medicinal plants that are widely used for medicinal purposes were used in this study. Publicly available sequences of four plastid markers were used for universal primer design. Species discrimination efficiency of the designed primers was evaluated as single and multi-locus analyses by using the primers sets. Methods HRM analysis was performed in triplicate on each of the 26 taxa to establish the Tm for each primer set (matK, rbcLA, rbcLB, rbcLC, rpoC1, and trnL). The shapes of the melting curves were analyzed to distinguish the different plant species. Bar-HRM species identification success rates were assessed for each single-locus as well as for multi-locus combinations to establish the optimal combination of primer sets. Results In single locus analysis the rpoC1 primer set gave the highest discrimination (58%), and in multi locus analysis this could be increased from 87% to 99% depending on the total number of regions included. Different combinations proved to be more or less effective at discrimination, depending on the genus or family examined. Conclusions Bar-HRM has proven to be a cost-effective and reliable method for the identification of species in this study of Thai medicinal plants, and results show an identification success rate of 99% among species in the test set.

Most people in the Swat valley of northwest Pakistan practice subsistence farming, supplementing their income by collecting and selling wild harvested plants for use in herbal medicine. Previous work showed that the collectors did not know the potential long-term impacts of collecting wild plants. We hypothesized that establishment of ex situ cultivation plots for these most valuable species would provide a sustainable alternative and lead to development of skills in agricultural production and marketing among participants. Swat valley farmers were helped to establish plots in four locations and taught to cultivate ten medicinal and aromatic plants (MAPs). In the first year, workshops were held on the financial benefits of growing MAPs for trade, criteria in selecting species, and strategies to improve MAP yield. In the second year, emphasis was placed on helping the farmers achieve a better price for their products by engaging them in discussions concerning criteria used in setting purchase prices. Seven of the ten cultivated MAPs yielded a better financial return than tomatoes (Solanum lycopersicum L.), the traditional cash crop in the area. Cultivating MAPs can yield a higher financial return than traditional cash crops, but a long-term assessment is necessary. Offering training in collection, preservation, and marketing can enhance the financial return and the long-term benefits of cultivation. Introduction of standardized production technology and appropriate post-harvest management has become a prime engine of growth for the economies of the subsistence farmers participating in our study, and is leading to better management and conservation practices for MAPs and the landscapes in which they grow. This is a post-peer-review, pre-copyedit version of an article published in Genetic Resources and Crop Evolution. The final authenticated version is available online at: http://dx.doi.org/10.1007/s10722-015-0346-z

Ethnopharmacological relevance Medicinal plant (MP) use during pregnancy is common in Africa and may have profound effects on both the mother and the developing foetus. A lack of overview complicates monitoring and regulating the use of MPs during pregnancy. Aim of the study This systematic review analyses prevalence of use of MPs during pregnancy, regional distribution, types and prevalence, MP properties, potential health risks, and consensus of MPs use, and suggests relevant measures to mitigate negative effects on pregnancy. Materials and methods A search was undertaken using a range of scientific databases (Medline, Embase, African Journals OnLine, Google Scholar and Biological Abstracts), non-governmental organisations, various African universities and regulatory websites for original published and unpublished studies that assess and indicate the prevalence of use of MPs during pregnancy in Africa. Additional articles were located by exploring pertinent bibliographies, and contacting experts. Results A total of 3659 MP-use studies were found, but only 303 articles received full-text assessment for eligibility and finally only 50 scientific papers were eligible for the systematic review. The prevalence of MP use by pregnant women varied widely from 2% to 100%. Twenty-eight studies (56%) specified one or more plant species used as MP during pregnancy. The major reasons for MP use were relief of nausea and vomiting during pregnancy (NVP), stimulation of labour, and facilitation of childbirth. The most commonly cited MP species were Zingiber officinale Roscoe, Allium sativum L. and Cucurbita pepo L. and these were used for relief of NVP, motion sickness and as a nutritional supplement. Route of administration was most commonly oral, and few adverse effects were reported. Conclusions The use of MPs among pregnant women in Africa is prevalent, and the most commonly used plant species are not known to have harmful foetal effects during pregnancy. However, many of the MP species are poorly studied and teratogenic effects cannot be ruled out. Collaboration between healthcare providers and traditional practitioners to inform about the safe use of MPs may promote safer pregnancies and better health for mothers and infants.

Nomenclature of plants in pharmacology can be presented by pharmaceutical names or scientific names in the form of Linnaean binomials. In this paper, positive and negative aspects of both systems are discussed in the context of the scientific nomenclatural framework and the systems' practical applicability. The Uppsala Monitoring Centre (UMC) runs the WHO Programme for International Drug Monitoring and is responsible for the WHO Adverse Drug Reaction (ADR) database that currently contains 3.6 million records. In order for the UMC to monitor pharmacovigilance through ADRs to herbal medicine products the following nomenclatural criteria are important: (i) the name should indicate only one species of plant; (ii) the source for this name must be authoritative; (iii) the name should indicate which part of the plant is used. Based on these criteria, the UMC investigated four options: (i) adopt main names used in recognised (inter-) national pharmacopoeias or authoritative publications; (ii) adopt option 1, but cite the publication for all names in abbreviated form; (iii) three-part pharmaceutical names consisting of Latinised part name plus Latinised genus name, plus Latinised specific epithet; (iv) scientific binomial names, optionally with author and plant part used. The UMC has chosen the latter option and will at its adoption utilise the scientific botanical nomenclature as defined by the International Code of Botanical Nomenclature. This decision satisfies all criteria set by the UMC and renders the necessity of creating a new system or upgrading an old inconsistent system obsolete. The UMC has also issued an extensive synonymy checklist of vernacular, pharmaceutical and scientific names for the herbals in the WHO ADR database. We strongly recommend the adoption of scientific names to denote plant ingredients in medicine.

Ethnopharmacological relevance In Tanzania, traditional medicine plays a significant role in health care and local economies based on the harvesting, trade and sale of medicinal plant products. The majority of this plant material is said to originate from wild sources, and both traditional healers and vendors are concerned about the increasing scarcity of certain species. Aim of the study A market survey of non-powdered, non-woody medicinal plants was conducted at Kariakoo Market in Dar es Salaam, the major hub for medicinal plant trade in Tanzania, to assess sustainability of traded herbal medicine. Materials and Methods For this study, fresh and dried herbs, seeds and fruits were collected and interviews were conducted to obtain information on vernacular names, preparation methods, monthly sales, uses and prices. Bundles of herbal medicine offered for sale were weighed and counted to calculate the value and volumes of daily stock at the market. Results A total of 71 medicinal plant products belonging to 62–67 different species from at least 41 different plant families were identified. We identified 45 plant products to species level, 20 products to genus level and four to family level. Plant species most encountered at the market were Suregada zanzibariensis, Myrothamnus flabellifolia and Sclerocarya birrea. The major use categories reported by the vendors were ritual purposes, digestive disorders and women's health. Annual sales are estimated to be in excess of 30 t and close to 200,000 USD, and trade in herbal medicine at Kariakoo Market provides subsistence income to many local vendors. Conclusions A large diversity of wild-harvested plant species is traded as medicinal products in Tanzania, including species listed on CITES Appendices. Identifying and monitoring temporal changes in availability per season and from year to year will reveal which species are most affected by this trade, and help relevant authorities in Tanzania to find alternative sources of income for dependent stakeholders and initiate targeted efforts to protect threatened plant species.

Pharmacovigilance of herbal medicines relies on the product label information regarding the ingredients and the adherence to good manufacturing practices along the commercialisation chain. Several studies have shown that substitution of plant species occurs in herbal medicines, and this in turn poses a challenge to herbal pharmacovigilance as adverse reactions might be due to adulterated or added ingredients. Authentication of constituents in herbal medicines using analytical chemistry methods can help detect contaminants and toxins, but are often limited or incapable of detecting the source of the contamination. Recent developments in molecular plant identification using DNA sequence data enable accurate identification of plant species from herbal medicines using defined DNA markers. Identification of multiple constituent species from compound herbal medicines using amplicon metabarcoding enables verification of labelled ingredients and detection of substituted, adulterated and added species. DNA barcoding is proving to be a powerful method to assess species composition in herbal medicines and has the potential to be used as a standard method in herbal pharmacovigilance research of adverse reactions to specific products.

BackgroundDifferences in regulatory policies between countries as well as a lack of appropriate standardized methods for authentication and quality control of herbal products lead to concerns over quality and safety.Echinaceaproducts are among the top selling herbal products in Europe and the United States with indications for broad range of ailments.PurposeThis study approached the need for a novel analytical strategy in authentication of herbal products.MethodsA combination of high performance thin layer chromatography (HPTLC) and DNA metabarcoding was employed. Fifty-threeEchinaceaherbal products marketed across Europe were tested to evaluate the accuracy of these methods in plant identification and their potential in detecting substitutes, adulterants and other unreported plant constituents.ResultsHPTLC provides a high resolution in detectingEchinaceaphytochemical target compounds, but does not offer information on the other species within the product. Alternatively, we showed that the limitation of HPTLC to detect non-targeted species can be overcome through the complementary use of DNA metabarcoding. Using DNA metabarcoding,Echinaceaspecies were detected in 34 out of the 38 retained products (89 %), but with a lack of discriminatory resolution at the species level due to the low level of molecular divergence within theEchinaceagenus. All herbal products showed considerable discrepancies between ingredients listed on the label and the ones detected using DNA metabarcoding registering an overall ingredient fidelity of 43 %.ConclusionThe results confirm that DNA metabarcoding can be used to test for the presence ofEchinaceaand simultaneously to detect other species present in even highly processed and multi-ingredient herbal products.AbbreviationsEMAEuropean Medicines AgencyBLASTbasic local alignment search toolFTIRFourier-transformed infrared spectroscopyIRinfrared spectrometryHPTLChigh performance thin layer chromatographymatKmaturase KMSmass spectrometryMOTUmolecular taxonomic unitnrITSnuclear ribosomal internal transcribed spacerPh.Eur.European PharmacopoeiarbcLribulose bisphosphate carboxylaseTLCthin-layer chromatographyUV-VISultraviolet-visible

Many herbal products have a long history of use, but there are increasing concerns over product efficacy, safety and quality in the wake of recent cases exposing discrepancies between labeling and constituents. When it comes to St. John’s wort (Hypericum perforatum L.) herbal products, there is limited oversight, frequent off-label use and insufficient monitoring of adverse drug reactions. In this study, we use amplicon metabarcoding (AMB) to authenticate 78 H. perforatum herbal products and evaluate its ability to detect substitution compared to standard methods using thin-layer chromatography (TLC) and high performance liquid chromatography coupled with mass spectrometry (HPLC-MS). Hypericum perforatum was detected in 68% of the products using AMB. Furthermore, AMB detected incongruence between constituent species and those listed on the label in all products. Neither TLC nor HPLC-MS could be used to unambiguously identify H. perforatum. They are accurate methods for authenticating presence of the target compounds, but have limited efficiency in detecting infrageneric substitution and do not yield any information on other plant ingredients in the products. Random post-marketing AMB of herbal products by regulatory agencies could raise awareness among consumers of substitution and would provide an incentive to manufacturers to increase quality control from raw ingredients to commercialized products.

Ethnopharmacological relevance What are the minimum methodological and conceptual requirements for an ethnopharmacological field study? How can the results of ethnopharmacological field studies be reported so that researchers with different backgrounds can draw on the results and develop new research questions and projects? And how should these field data be presented to get accepted in a scientific journal such as the Journal of Ethnopharmacology? The objective of this commentary is to create a reference that covers the basic standards necessary during planning, conducting and reporting of field research. Materials and methods We focus on conducting and reporting ethnopharmacological field studies on medicinal plants or materia medica and associated knowledge of a specific people or region. The article highlights the most frequent problems and pitfalls, and draws on published literature, fieldwork experience, and extensive insights from peer-review of field studies. Results Research needs to be ethical and legal, and follow local and national regulations. Primary ethnopharmacological field data need to be collected and presented in a transparent and comprehensible way. In short this includes: 1) Relevant and concise research questions, 2) Thorough literature study encompassing all available information on the study site from different disciplines, 3) Appropriate methods to answer the research questions, 4) Proper plant use documentation, unambiguously linked to voucher specimens, and 5) Qualitative and quantitative analyses of the collected data, the latter relying on use-reports as basic units. Conclusion Although not exhaustive, we provide an overview of the necessary main issues to consider for field research and data reporting including a list of minimal standards and recommendations for best practices. For methodological details and how to correctly apply specific methods, we refer to further reading of suggested textbooks and methods manuals. Weckerle, Caroline S., et al. "Recommended standards for conducting and reporting ethnopharmacological field studies." Journal of ethnopharmacology 210 (2018): 125-132. © 2018. This manuscript version is made available under the CC-BY-NC-ND 4.0 license.

In eastern Mediterranean countries orchids continue to be collected from the wild for the production of salep, a beverage made of dried orchid tubers. In this study we used nrITS1 and nrITS2 DNA metabarcoding to identify orchid and other plant species present in 55 commercial salep products purchased in Iran, Turkey, Greece and Germany. Thirty samples yielded a total of 161 plant taxa, and 13 products (43%) contained orchid species and these belonged to 10 terrestrial species with tuberous roots. Another 70% contained the substitute ingredient Cyamopsis tetraganoloba (Guar). DNA metabarcoding using the barcoding markers nrITS1 and nrITS2 shows the potential of these markers and approach for identification of species used in salep products. The analysis of interspecific genetic distances between sequences of these markers for the most common salep orchid genera shows that species level identifications can be made with a high level of confidence. Understanding the species diversity and provenance of salep orchid tubers will enable the chain of commercialization of endangered species to be traced back to the harvesters and their natural habitats, and thus allow for targeted efforts to protect or sustainably use wild populations of these orchids.

Species Substitution in Medicinal Roots and Possible Implications for Toxicity of Herbal Remedies in Morocco. Herbal medicine is an integral part of health care in Morocco and is widely used by Moroccans. However, the efficacy and safety of traditional plant-based medicine in Morocco is threatened by insufficient knowledge about practices of adulteration and substitution. These issues are of particular importance when subterranean plant parts are employed. A combination of qualitative and quantitative methods was used to identify where and why confusion (accidental substitution) and intentional substitution occurs in root-based medicines sold in Marrakech. Additionally we examined local perceptions of the toxicological risks posed by substitution. We recorded a total of 20 species (34 unique cases) of roots for which substitution or confusion was reported by either herbalists or collectors. Substitution or confusion occurred in 54.6 % of the 33 most commonly sold medicinal roots, with herbalists reporting substitution in more species than collectors. Collectors and herbalists cited poor availability of roots (in part due to overexploitation of wild resources), high demand, high prices, and lack of knowledge as factors driving substitution and confusion. Roots for which substitution was reported were significantly more difficult for herbalists to identify. Moreover, profit was higher for roots for which intentional substitution was reported. Despite the detailed knowledge held by many herbalists and a long tradition of use of herbal medicine in Morocco, doctors and pharmacists had dismissive attitudes towards traditional medicine and expressed concern about both efficacy and safety of medicinal plant use. Given the high rates of substitution and confusion documented by this study, there is an urgent need for appropriate regulation of herbal remedies in Morocco, a conclusion embraced by herbalists and Western-trained medical practitioners alike.

The traditional use of medicinal plants for the treatment of human and livestock ailments is important to indigenous communities in the northern parts of Pakistan, and considered to be a valuable local biological and sociocultural heritage. The aim of this study was to obtain a detailed inventory of medicinal plant use and preparation among Kalasha, Ismaeli and Sunni groups.Semi-structured group and individual interviews were carried out with men and women of different age groups that identified themselves as being Kalasha, Ismaeli or Sunni. Interviews were followed up by field visits to collect herbarium vouchers and record in greater detail the exact methods of harvesting, preparation and use on medicinal plants.A total of 76 species were recorded for treatment of various diseases. The Kalasha, Ismaili and Sunni ethnic groups have similar medicinal floras, but show striking differences in plant use. Our comparative survey shows that out of all species reported in this study, only 13 species have been reported previously from Chitral District.Indigenous knowledge of folk medicine is intricately linked to local culture, religion and history. Any short study can only scratch the surface of this intricate system, but provide an insight into the critical importance of medicinal plants for local livelihoods and the important role these play in health care systems. There is a great need to assess and properly manage the production potential of medicinal plants to ensure sustainable supply of these species for local use and subsistence trade.

Pelargonium (Pelargonium sidoides DC and P. reniforme Curtis) is reported to have immune modulating properties and antibacterial activity, and Pelargonium extracts have been used for the treatment of respiratory tract and gastrointestinal infections. Introduced in the early 1980s in Germany, Umckaloabo® (ISO Arzneimittel), an ethanolic extract of the roots of P. sidoides and P. reniforme, was the first Pelargonium-derived product to be commonly used in a country in the EU. According to the Umckaloabo® product information, this extract has no known adverse effects. However, there is a theoretical risk of interactions with anticoagulants such as warfarin, and antiplatelet drugs, such as aspirin (acetylsalicylic acid). To date, the Uppsala Monitoring Centre has, through the WHO international pharmacovigilance programme, received 34 case reports of allergic reactions suspected to be associated with the use of Pelargonium extract, all originating from Germany. In a number of these reports, the description and timing of the event was indicative of an acute Coombs and Gell Type I hypersensitivity reaction; two of these patients needed treatment for circulatory failure. So far, the experience of such reactions is limited to Germany. Since Pelargonium-containing herbal products have recently been approved in a number of other countries, the possibility of the occurrence of allergic reactions has become of more general interest and further information regarding these products is needed.

Background: An accessory to modern developing economies includes a shift from traditional, laborious lifestyles and cuisine to more sedentary careers, recreation and convenience-based foodstuffs. Similar changes in the developed western world have led to harmful health consequences. Minimization of this effect in current transitional cultures could be met by placing value on the maintenance of heritage-rich food. Vitally important to this is the preservation and dissemination of knowledge of these traditional foods. Here, we investigate the history and functionality of a traditional rice snack cooked in Nepenthes pitchers, one of the most iconic and recognizable plants in the rapidly growing economic environment of Southeast Asia. Methods: Social media was combined with traditional ethnobotanical surveys to conduct investigations throughout Malaysian Borneo. Interviews were conducted with 25 market customers, vendors and participants from various ethnical groups with an in-depth knowledge of glutinous rice cooked in pitcher plants. The acidity of pitcher fluid was measured during experimental cooking to analyze possible chemical avenues that might contribute to rice stickiness. Results: Participants identifying the snack were almost all (96%) from indigenous Bidayuh or Kadazandusun tribal decent. They prepare glutinous rice inside pitcher traps for tradition, vessel functionality and because they thought it added fragrance and taste to the rice. The pH and chemical activity of traps analyzed suggest there is no corresponding effect on rice consistency. Harvest of pitchers does not appear to decrease the number of plants in local populations. Conclusions: The tradition of cooking glutinous rice snacks in pitcher plants, or peruik kera in Malay, likely carries from a time when cooking vessels were more limited, and persists only faintly in tribal culture today because of value placed on maintaining cultural heritage. Social media proved a valuable tool in our research for locating research areas and in interviewing respondents, and we endorse its further use in ethnobotanical investigations. Our gathered data urges for the preservation of sustainable, tribal plant use for the prosperity of both health and culture.

Medicinal plants are used as a popular alternative to synthetic drugs, both in developed and developing countries. The economic importance of the herbal and natural supplement industry is increasing every year. As the herbal industry grows, consumer safety is one issue that cannot be overlooked. Herbal products in Thai local markets are commonly sold without packaging or labels. Plant powders are stored in large bags or boxes, and therefore buying local herbal products poses a high risk of acquiring counterfeited, substituted and/or adulterated products. Due to these issues, a reliable method to authenticate products is needed. Here DNA barcoding was used in combination with High Resolution Melting analysis (Bar-HRM) to authenticate three medicinal Acanthaceae species (Acanthus ebracteatus, Andrographis paniculata and Rhinacanthus nasutus) commonly used in Thailand. The rbcL barcode was selected for use in primers design for HRM analysis to produce standard melting profiles of the selected species. Melting data from the HRM assay using the designed rbcL primers showed that the three chosen species could be distinguished from each other. HRM curves of all fifteen test samples indicated that three of tested products did not contain the indicated species. Two closely related species (A. paniculata and R. nasutus), which have a high level of morphological similarity, were interchanged with one another in three tested products. Incorrect information on packaging and labels of the tested herbal products was the cause of the results shown here. Morphological similarity among the species of interest also hindered the collection process. The Bar-HRM method developed here proved useful in aiding in the identification and authentication of herbal species in processed samples. In the future, species authentication through Bar-HRM could be used to promote consumer trust, as well as raising the quality of herbal products.

Mapping ethnomedicinal plants and associated indigenous knowledge of folk medicines can provide a comprehensive overview of individual herbs employed in health care. Reliance on medicinal plants in remote parts of northern Pakistan is high, especially among women, but no research has investigated specifically which plants are used. This study investigated indigenous knowledge of folk medicines among tribal minorities in selected sites in upper Swat, Buner and Chitral Districts in Khyber Pakhtunkhwa Province.Interviews were conducted with gender-specific focus groups using questionnaires and standardized data sheets, followed by forest walks in each of the visited areas. General medicinal herb use, preparations, storage, marketing and collection habits for each gender group were ascertained from the questionnaires.In total 168 women and 390 men were interviewed and provided information on 127 different shared medicinal species. Species use consensus among the informants ranged from 2.3% to 83.3%, with Cynodon dactylon, Avena sativa, Celtis australis, Datura stramonium, Solanum nigrum, Skimmia laureola, Spiraea nervosa, Ziziphus jujuba, Rumex hastatus, Plantago lanceolata, Lathyrus aphaca and Ficus palmata having the highest reported consensus. The survey also revealed that a number of medicinal species were exploited by the community for both marketing and personal use, and many of these species were reported as being rare, vulnerable or even endangered.The results revealed that women in all the three districts were important custodians of medicinal plant knowledge, but elder women in general and the women from Buner district in particular had a superior understanding of folk medicine. The forest walks revealed that women׳s traditional medicinal knowledge was based on a more limited diversity of plant species. People in tribal communities have an expressed interest in learning efficient techniques for medicinal plant collection, preparation, storage and cultivation advice, and to learn more about the potential of marketing medicinal herbs and ways to reach local market centers. Education and awareness were considered to be essential for improved health care and successful marketing.

The field of molecular biology has brought many innovations to biodiversity research, with the emerging field of DNA barcoding, which is having a growing impact on plant identification. In DNA barcoding, standard molecular markers are used for species identification of material that has hitherto been hard or impossible to identify, such as sterile plant vouchers, plant parts and even processed herbal products. Widespread use of DNA barcoding for plant identification faces challenges such as the selection of suitable markers that are both variable and easy to amplify, and the exhaustive presence of these markers for species in public nucleotide repositories. In recent years, researchers have successfully shown the power and accuracy of plant barcoding by creating molecular identification systems for various purposes, such as the identification of specific medicinal plant species susceptible to adulteration, overharvested and endangered taxa, and monitoring of cross-border trade in CITES-listed species. As markers, nucleotide repositories and sequencing technology improve, these methods will become ubiquitous and will facilitate the identification of products derived from wild-harvested medicinal plants and the enforcement of the Convention on Biological Diversity Nagoya protocol on Access and Benefit Sharing.

Ethnopharmacological relevance This is an extensive review of plants used traditionally for women's healthcare in Southeast Asia and surrounding countries. Medicinal plants have a significant role in women's healthcare in many rural areas of the world. Plants with numerous efficacious observations have historically been used as a starting point in the development of new drugs, and a large percentage of modern pharmaceuticals have been derived from medicinal plants. Materials and methods A review was conducted for all plant use mentioned specifically for female healthcare, such as medicine to increase fertility, induce menstruation or abortion, ease pregnancy and parturition, reduce menstrual bleeding and postpartum hemorrhage, alleviate menstrual, parturition and postpartum pain, increase or inhibit lactation, and treat mastitis and uterine prolapse, in 200 studies focusing on medicinal plant use, either general studies or studies focusing specifically on women's healthcare. Results Nearly 2000 different plant species are reported to be used in over 5000 combinations. Most common are Achyranthes aspera, Artemisia vulgaris, Blumea balsamifera, Carica papaya, Curcuma longa, Hibiscus rosa-sinensis, Leonurus japonicus, Psidium guajava and Ricinus communis, and each of these species had been reported in more than 10 different scientific articles. Conclusions This review provides a basis for traditional plant use in women's healthcare, and these species can be used as the starting point in the discovery of new drugs.

Background Activities and diet during the postpartum period are culturally dictated in many Southeast Asian cultures, and a period of confinement is observed. Plants play an important role in recovery during the postpartum period in diet and traditional medicine. Little is known of the Kry, a small ethnic group whose language was recently described, concerning its traditions and use of plants during pregnancy, parturition, postpartum recovery and infant healthcare. This research aims to study those traditions and identify medicinal plant use. Methods Data were collected in the 3 different Kry villages in Khammouane province, Lao PDR, through group and individual interviews with women by female interviewers. Results A total of 49 different plant species are used in women's healthcare. Plant use is culturally different from the neighboring Brou and Saek ethnic groups. Menstruation, delivery and postpartum recovery take place in separate, purpose-built, huts and a complex system of spatial restrictions is observed. Conclusions Traditions surrounding childbirth are diverse and have been strictly observed, but are undergoing a shift towards those from neighboring ethnic groups, the Brou and Saek. Medicinal plant use to facilitate childbirth, alleviate menstruation problems, assist recovery after miscarriage, mitigate postpartum haemorrhage, aid postpartum recovery, and for use in infant care, is more common than previously reported (49 species instead of 14). The wealth of novel insights into plant use and preparation will help to understand culturally important practices such as traditional delivery, spatial taboos, confinement and dietary restrictions, and their potential in modern healthcare.

Background Fundamental in traditional postpartum recovery in Lao PDR is the use of hotbeds, mother roasting, steam sauna and steam baths. During these treatments medicinal plants play a crucial role, but little has been published about how the treatments are carried out precisely, which species are used, the medicinal properties of these species, and the medicinal efficacy of their chemical constituents. Methods Sixty-five interviews, in 15 rural villages, with women of 4 different ethnic groups were conducted to survey confinement rituals, and postpartum plant use and salience. Essential oils from the main species used were extracted using steam distillation and the main chemical constituents characterized using gas chromatography-mass spectrometry (GC-MS). Results A total of 10 different species were used by three or more of the ethnic groups included in this study. All species were used in steam sauna and bath, but only 3 species were used in hotbed and mother roasting. Essential oils of Amomum villosum, Amomum microcarpum and Blumea balsamifera were found to contain significant amounts of the following terpenes: β-pinene, camphor, bornyl acetate, borneol, linalool, D-limonene, fenchone, terpinen-4-ol and α-terpinene. Conclusions Many of these terpenes have documented antimicrobial and analgesic properties, and some have also synergistic interactions with other terpenes. The mode of application in hotbed and mother roasting differs from the documented mechanisms of action of these terpenes. Plants in these two practices are likely to serve mainly hygienic purposes, by segregating the mother from infection sources such as beds, mats, stools, cloth and towels. Steam sauna medicinal plant use through inhalation of essential oils vapors can possibly have medicinal efficacy, but is unlikely to alleviate the ailments commonly encountered during postpartum convalescence. Steam sauna medicinal plant use through dermal condensation of essential oils, and steam bath cleansing of the perineal area is possibly a pragmatic use of the reported medicinal plants, as terpene constituents have documented antimicrobial, analgesic and anti-inflammatory properties.