Wild Mushrooms Research Articles

Exploring time-killing and biofilm inhibition potential of bioactive proteins extracted from two varieties of Pleurotus ostreatus

IntroductionDental caries, caused by oral microbial pathogens, are a global health concern, further exacerbated by the presence of methicillin-resistant Staphylococcus aureus (MRSA). Bioactive proteins and peptides (BAPs) exhibit potent antimicrobial properties, targeting multiple cellular mechanisms within pathogens, reducing the likelihood of resistance development. Given the antimicrobial potential of BAPs, this study aimed to compare the efficacy of BAPs extracted from cultivated (Pleurotus ostreatus, PoC) and wild (Pleurotus ostreatus, PoW) mushrooms against pathogens responsible for dental caries.MethodsBAPs were extracted from both PoC and PoW using a TCA-acetone method. Antimicrobial activities were tested against seven bacteria and one fungus using agar well diffusion and MIC determination. Antibiofilm activity was assessed via modified CV assay, while DPPH and erythrocyte lysis tests evaluated free radical scavenging.ResultsPoC showed superior antimicrobial efficacy, with lower MIC and MBC values, and disrupted biofilm integrity at increasing concentrations. PoW exhibited better antioxidant activity with higher DPPH scavenging, though its antimicrobial efficacy was slightly lower than PoC.DiscussionBoth PoC and PoW BAPs inhibited dental pathogens, with PoC showing stronger inhibition against MRSA and nystatin-resistant Candida albicans. This suggests BAPs may target additional cellular mechanisms beyond membranes, PBPs, and ergosterols. Despite PoW’s stronger antioxidant properties, both BAPs had comparable antibiofilm activity. These findings suggest complementary actions of BAPs from PoC and PoW both, in treating dental caries, offering broad-spectrum antimicrobial and antioxidant benefits.

Exploring the Relationship Between Time Series of Sentinel-1 Interferometric Coherence Data and Wild Edible Mushroom Yields in Mediterranean Forests

Edible wild mushrooms constitute a valuable marketable non-wood forest product with high relevance worldwide. There is growing interest in developing tools for estimation of mushroom yields and to evaluate the effects that global change may have on them. Remote sensing is a powerful technology for characterization of forest structure and condition, both essential factors in triggering mushroom production, together with meteo-climatic factors. In this work, we explore options to apply synthetic aperture radar (SAR) data from C-band Sentinel-1 to characterize, at the plot level, wild mushroom productive forests in the Mediterranean region, which provide saprotroph and ectomycorrhizal mushrooms. Seventeen permanent plots with mushroom yield data collected weekly during the productive season are characterized with dense time series of Sentinel-1 backscatter intensity (VV and VH polarizations) and 6-day interval interferometric VV coherence during the 2018–2021 period. Weekly-regularized series of SAR data are decomposed with a LOESS approach into trend, seasonality, and remainder. Trends are explored with the Theil-Sen test, and periodicity is characterized by the Discrete Fast Fourier transform. Seasonal patterns of SAR time-series are described and related to mycorrhizal and saprotroph guilds separately. Our results indicate that time series of interferometric coherence show cyclic patterns which might be related with annual mushroom yields and may constitute an indicator of triggering factors in mushroom production, whereas backscatter intensity is strongly correlated with precipitation, making noisy signals without a clear interpretable pattern. Exploring the potential of remotely sensed data for prediction and quantification of mushroom yields contributes to improve our understanding of fungal biological cycles and opens new ways to develop tools that improve its sustainable, efficient, and effective management.

Characterization of Important Odorants in Dried Lobster Mushrooms.

Lobster mushrooms, a type of wild edible mushroom that primarily grows in North America, have a distinctive aroma resembling seafood and are highly desired by fine dining establishments and food enthusiasts. Lobster mushrooms have a short collection season; therefore, they are usually dried for a year-round sale. Owing to their rising popularity, lobster mushrooms have become one of the most expensive mushroom species in the dried mushroom market. Herein, 35 odorants were identified from dried lobster mushrooms using solvent assisted flavor evaporation (SAFE) and aroma extract dilution analysis (AEDA). Fourteen odorants were quantitated using stable isotope dilution assays (SIDA), and their odor activity values (OAVs) were calculated. Eleven of the quantitated odorants exhibited OAVs of ≥1. Some notable odorants with relatively high OAVs included 2-acetyl-1-pyrroline (OAV 730), γ-nonalactone (OAV 410), 1-octen-3-one (OAV 180), and sotolon (OAV 170). The organohalogen molecule 2,6-dichlorophenol was determined to be key to the seafood aroma character of the mushrooms. An aroma simulation model formulated based on the quantitation data closely matched the aroma profile of the dried mushrooms. This study lays the groundwork for future studies aimed at understanding the effect of processing on lobster mushroom aroma chemistry and the biotechnological production of natural seafood flavors.

Purification and characterisation of laccase from the medicinal wild mushroom Coriolus brevis

Purification and characterisation of laccase from the medicinal wild mushroom Coriolus brevis

Sensory Characteristics and Volatile Organic Compound Profile of Wild Edible Mushrooms from Patagonia, Argentina.

The Andean-Patagonian forests of South America offer a great variety of wild edible mushrooms, many with ancestral use and others linked to new mycogastronomic offers. However, their sensory properties and detailed characterizations have not yet been deeply explored and described, nor have their alterations due to cold storage. The aims of this work were to perform a sensory characterization through a trained panel evaluation, perform target volatile compounds analysis and evaluate post-harvest preservation methods effects on nine species of wild edible mushrooms with different trophic habits (Cortinarius magellanicus, Panus dusenii, Fistulina antarctica, F. endoxantha, Gloeosoma vitellinum, Grifola gargal, Lepista nuda, Ramaria patagonica, and Cyttaria hariotii). The sensory description of dehydrated specimens through quantitative descriptive analysis showed that panelists were a significant source of variation; F. antarctica and R. patagonica registered distinct sweet flavor/spice odor and wood/sweet flavor, respectively, and different textures. Refrigeration produced a rapid loss of sensory characteristics, whereas freezer conservation satisfactorily maintained the characteristics in F. anctartica, R. patagonica, G. vitellinum, and C. hariotti for at least four months. A total of 60 target volatile organic compounds were detected, corresponding to grass, mushroom, alkane, and pungent odors in F. anctartica, R. patagonica, and G. vitellinum. The detailed sensory characterization and post-harvest conservation options of these novel products constitute crucial information to promote their sustainable use and local development through innovative activities linked to tourism, such as mushroom gastronomy and mycotourism.

Rapid and non-invasive estimation of total phenol content and species identification in dried wild edible bolete using FT-NIR spectroscopy

The market for dried wild edible mushrooms is characterized by quality discrepancies and species disorganization, which are a matter of concern. The feasibility of using fourier transform near-infrared (FT-NIR) spectroscopy with chemometrics to predict the total phenol content and identify species was investigated in dried bolete. For the determination of total phenolic content, FT-NIR spectral acquisition, and two-dimensional correlation spectroscopy (2DCOS) image acquisition, five common species of dried edible bolete (n = 144) were utilized. The results showed that partial least squares regression (PLSR) combined with Fourier transform near-infrared spectroscopy could predict the total phenol content of dried boletes, with the best prediction coefficient of determination (R2p) = 0.79. The partial least squares discriminant analysis (PLS-DA) model accurately identified Phlebopus portentosus, with Matthews correlation coefficient (MCC), Precision (PRC), Sensitivity (SEN), and Specificity (SPE) all being 1. The support vector machines (SVM) model is performed optimally to identify processing edible bolete (Lanmaoa asiatica) with an accuracy of 100 % in the test set. 2DCOS images combined with the residual convolutional neural networks (ResNet) model demonstrated the feasibility of FT-NIR full spectral bands (10,000–4,000 cm−1) and characteristic spectral bands (6,500–4,000 cm−1) for species identification of boletes. The method applies to the case of consistent or inconsistent sample sizes between groups, with an accuracy of 1.00 for both the training and test sets. The study serves as a rapid, non-invasive, and convenient method for real-time evaluation of the quality of dried edible bolete in the market.

Harvest and trade of wild edible russula griseocarnosa in North Vietnam

This study examines the harvest and trade of wild edible Russula griseocarnosa in north Vietnam. Russula griseocarnosa is an edible forest mushroom, but there is no reliable information on harvesting practices and trade in this species in Vietnam. This study aims to provide ethnomycological and marketing knowledge for this mushroom. Semi-structured interviews were conducted with mushroom collectors and traders over 2 years; harvest yields were obtained from local officials; and policy settings were sought from forest managers. About 85% of mushroom collectors were women of the Dao, Lo Ursula, Muong, Nung, Mong, San Chi, San Diu, and Tay ethnicities. The collecting season was bimodal, with the wet season beginning (May) and ending (September). The traders were Hoa, Kinh, Nung and Tay, and 57% were women. Fresh mushrooms sold for 9.1-10.5 US$/kg on dry days and 7.0-7.9 US$/kg on wet days. The best grade of dried product fetched 70.4-83.3 US$/kg. A number of interim policies have been established to encourage local people to protect forests and to help create a sustainable edible wild mushroom market. Our study provides the first detailed account of the role of edible wild mushrooms in northeast Vietnam, and this will inform the co-development of rural livelihood and sustainable forest management plans.

Diversity and Ethnomycological Knowledge of Wild Mushrooms Consumed by Indigenous People Living Near the Botambi Classified Forest in the Central African Republic

Diversity and Ethnomycological Knowledge of Wild Mushrooms Consumed by Indigenous People Living Near the Botambi Classified Forest in the Central African Republic

Real-time Deep Learning Based Mobile Application for Detecting Edible Fungi: Mushapp

Mushroom consumption and wild mushroom gathering are increasing in our country and in the world. Mushroom poisoning has an important place in food poisoning cases. Mushroom poisoning accounts for approximately 7% of poisoning cases in adults. Mushroom collection and consumption is common in many regions of our country. In this study, a deep learning based mobile application was developed to reduce the incidence of mushroom poisoning by taking a photo of a mushroom and determining the type and toxicity of the mushroom from the photo. This mobile application is called MushAPP. In the first phase of the study, 5150 mushroom images of 20 mushroom species were used to create the dataset. The dataset was then pre-processed and converted into a format that can be used by the deep learning algorithm. The mobile application side of the project was developed in Android Studio IDE environment. An artificial intelligence model was integrated into the designed mobile application. In the application, the type and toxicity status of the mushroom viewed from the mobile device camera are determined and presented to the user. The research findings were analyzed and it was determined that the accuracy rate of the application in detecting the mushroom species was 99.8%.

Nutritional, Phytochemical and Therapeutic Attributes of Edible Wild Mushrooms as Influenced by Substrates in Humid Tropical Environment

Nutritional, Phytochemical and Therapeutic Attributes of Edible Wild Mushrooms as Influenced by Substrates in Humid Tropical Environment

Detection and quantification of Amatoxin in wild mushrooms from North-East India using HPLC-PDA method for food safety purposes

Misidentification and ingestion of poisonous mushrooms pose significant threats to food safety, particularly in Mizoram, India, where over ten fatalities due to mushroom poisoning have been reported in the past decade (2013-2023). This study aimed to address this critical issue by identifying and quantifying the cause of death due to consumption of wild mushroom from Champhai district, Mizoram, India and to test the reliability of HPLC-PDA for detection and quantification of amatoxins. HPLC-PDA confirmed the presence of α-amanitin in Amanita virosa and Amanita bisporigera in the samples. α-amanitin is a water-soluble, heat-stable, and highly toxic cyclic octapeptide present in the genus Amanita, which includes Amanita phalloides, Amanita verna, and Amanita virosa. Amanitin cytotoxicity arises from the inhibition of RNA polymerases, namely RNA polymerase II, which obstructs mRNA production in kidney and liver cells. Validation of the method demonstrated good precision and accuracy, with LOD and LOQ values of 88 ng g-1 and 210 ng g-1, respectively. The method was successfully applied to quantify α-amanitin in ten wild mushroom samples, revealing its presence only in Amanita virosa (1.17 mg g-1) and Amanita bisporigera (1.91 mg g-1) species. These findings underscore the importance of accurate α-amanitin detection methods in ensuring food safety and public health, particularly in regions prone to mushroom poisoning incidents. It is noteworthy that this study marks the initial exploration for detection and quantification of α-amanitin from poisonous mushrooms found in the wild regions of Champhai district in Mizoram, representing the first report of such in the area.

LC-MS/MS Profiling of the Poisonous Wild Mushroom, Entoloma mastoideum, (Entolomataceae, Basidiomycota) in Sabah (Northern Borneo), Malaysia

In Malaysia, mushroom poisoning poses a significant public health concern, particularly as local communities frequently consume wild mushrooms without fully recognizing the potential dangers they present. Ingestion of wild mushrooms, which resemble edible species but contain deadly chemicals, is the main cause of poisoning risk. The genus Entoloma (Fr.) P. Kumm is dispersed worldwide, with over 1,500 species recorded. Entoloma mastoideum, a poisonous mushroom, was recorded for the first time at Kota Marudu district, Sabah, Malaysia, where ten cases of mushroom poisoning outbreaks were recorded during the rainy season of October 2019. The morphological characters of the specimens were documented and their microscopic features were analyzed using compound microscope and scanning electron microscope. A phylogenetic tree was constructed using Maximum likelihood and Bayesian analysis. Chemical profiling of the poisonous mushroom specimen was done via liquid chromatograph mass spectrometry (LC-MS/MS) to identify toxic metabolites. The phylogenetic analysis showed that the Bornean E. mastoideum is closely related to the Chinese isolates (100% BS/1.0 PP). LC-MS/MS profiling detected a total of 162 metabolites that were classified into ten general groups, where several toxic compounds were detected amongst the aromatics, essential amino acids, and fatty acid derivatives. The toxic compounds identified in the mushroom extract, including amino acid derivatives such as 3,4,5,6-tetrahydroxyynorleucine, valpromide, and betaine, have been reported to cause neurotoxicity, cardiotoxicity, liver damage, and gastrointestinal harm. The presence of these toxic compounds underscores the need for caution when consuming wild mushrooms. Further research on poisonous Entoloma species is vital for developing accurate identification methods and understanding their toxic potential. This knowledge is essential for enhancing public awareness, preventing mushroom poisoning incidents and safeguarding public health.

Prediction of the postharvest quality of Boletus wild mushrooms stored with mesoporous silica nanoparticles antibacterial film using Long Short-Term Memory model combined with the Northern Goshawk Optimization (NGO-LSTM)

This study aimed to address the challenge of extending the shelf life of Boletus wild mushrooms, which are prone to environmental and microbial contamination. An antibacterial film composed of polylactic acid (PLA) and mesoporous silica nanoparticles loaded with citral (CMP film) was developed for this purpose. Fifteen quality indices were assessed, and the data were integrated using AHP and TOPSIS to evaluate the film's efficacy. The CMP film effectively maintained the quality of mushroom over time. Additionally, a Nonlinear Global Optimization-Long Short-Term Memory (NGO-LSTM) model was employed to predict storage quality, using seven highly correlated quality indicators. The model achieved a high predictive accuracy, with the R2 exceeding 0.999. This study presents a novel packaging solution and a predictive model that together enhance the storage and quality control of Boletus wild mushrooms.

The Situation of Counterfeited and Mislabeled Commercialized Edible Mushrooms in China and the Development of Possible Controls

Edible mushroom products, encompassing both cultivated and wild varieties, are highly favored by consumers due to their rich nutritional profiles, including significant levels of proteins and amino acids. These mushrooms have extensive applications across the food, pharmaceutical, and cosmetic industries, making the edible mushroom industry a vital component of global poverty alleviation efforts. Taking China as an example, the country produces over 45 million tons of edible mushrooms annually, accounting for 94.01% of the world’s total production, thereby establishing itself as the leading global producer of edible mushrooms. However, alongside the rapid expansion of this industry, concerns have emerged regarding counterfeit products and incidents of poisoning resulting from the consumption of toxic wild mushrooms. As follows, to advance the development and integrity of the mushroom production and processing industry: (1) This study presents the situation of counterfeit edible mushrooms and elucidates the factors contributing to the production of fraudulent products from both subjective and non-subjective perspectives. (2) We provide a detailed introduction to 22 varieties of freshly cultivated edible mushrooms and commonly encountered wild edible mushrooms in the Chinese consumer market, proposing the application of DNA barcoding, environmental DNA analysis, and other technologies for the future authentication of counterfeit mushroom products. (3) Concurrently, we present an overview of mushroom poisoning incidents in China from 2010 to 2023, emphasizing the challenges in mitigating the risks associated with wild mushroom consumption and preventing food poisoning, thereby necessitating heightened consumer caution. (4) Finally, we offer four recommendations aimed at ensuring the healthy, stable, and sustainable growth of the edible mushroom industry.

Characterization of volatile profiles and markers prediction of eleven popular edible boletes using SDE-GC–MS and FT-NIR combined with chemometric analysis

Wild edible boletes mushrooms are regarded as a delicacy in many countries and regions due to their rich nutritional contents and strong aromatic compounds. This study aimed to identify 445 samples of 11 boletes species collected from Yunnan and Sichuan provinces through molecular analysis. Using simultaneous distillation–extraction (SDE) combined with gas chromatography-mass spectrometry (GC–MS), 97 volatile compounds were identified. Chemometric methods were then applied to analyze the heterogeneity of these volatile compounds among the different species. The results showed that, 22 and 21 volatile compounds were selected using variable importance in projection (VIP > 1) and relative odor activity values (ROAV > 0.1), respectively. Partial least squares discrimnatint analysis (PLS-DA) was then employed to develop pattern recognition models for 11 species, which demonstrated strong identification performance. Furthermore, correlation heat maps, volcano plots, and Fisher linear discriminant analysis identified five volatile organic compounds, including methyl (9E)-9-octadecenoate, 2, 6-dimethylpyrazine, 1-decen-3-one, furfural, and methional as markers for distinguishing 11 boletes species. Ultimately, the rapid content prediction models of partial least squares regression (PLSR) were established by combining Fourier Transform Near-Infrared Spectroscopy (FT-NIR) with the concentrations of these five marker compounds. These findings provide a methodological strategy for the effective species identification of wild edible mushrooms and the rapid prediction of their characteristic aroma compounds.

Diversity of wild mushrooms and ethnomycological studies in Mizoram

Diversity of wild mushrooms and ethnomycological studies in Mizoram

Организационные аспекты заготовки и переработки дикоросов в Пермском крае

The scientific article contains a review of scientific literature on the issues of economic development of collection and processing of wild plants, it presents the opinions of scientists on the great potential of the market of wild berries, mushrooms and medicinal herbs, that this type of activity can be one of the sources of income for the local population and a promising activity, especially for the population of small towns and rural areas. It is noted that the business of harvesting wild plants includes the following stages: collection of wild plants, procurement and processing (including primary and deep). Attention is drawn to the fact that the level of pre-sale preparation has a significant impact on the price, helps to ensure year-round sale of harvested berries, mushrooms and medicinal herbs. Wild plants in the form of berries and mushrooms can be sold fresh and undergo deep processing (freezing, drying, canning). Wild plants in the form of medicinal herbs are more in need of drying, which will significantly increase the shelf life. The business of harvesting wild plants differs in the level of initial investment: minimal investment is possible when selling wild plants fresh, for deep processing, special equipment for drying, freezing and preservation is needed. It is noted that the main consumers of frozen mushrooms and berries are private confectioners and pastry shops, dried berries - producers of cereals, canned mushrooms and berries - educational institutions and children's camps, catering establishments, stores. The profitability of the business justifies the presence of competitors in the wild plant market.

Research on the Wild Mushroom Recognition Method Based on Transformer and the Multi-Scale Feature Fusion Compact Bilinear Neural Network

Wild mushrooms are popular for their taste and nutritional value; however, non-experts often struggle to distinguish between toxic and non-toxic species when foraging in the wild, potentially leading to poisoning incidents. To address this issue, this study proposes a compact bilinear neural network method based on Transformer and multi-scale feature fusion. The method utilizes a dual-stream structure that integrates multiple feature extractors, enhancing the comprehensiveness of image information capture. Additionally, bottleneck attention and efficient multi-scale attention modules are embedded to effectively capture multi-scale features while maintaining low computational costs. By employing a compact bilinear pooling module, the model achieves high-order feature interactions, reducing the number of parameters without compromising performance. Experimental results demonstrate that the proposed method achieves an accuracy of 98.03%, outperforming existing comparative methods. This proves the superior recognition performance of the model, making it more reliable in distinguishing wild mushrooms while capturing key information from multiple dimensions, enabling it to better handle complex scenarios. Furthermore, the development of public-facing identification tools based on this method could help reduce the risk of poisoning incidents. Building on these findings, the study suggests strengthening the research and development of digital agricultural technologies, promoting the application of intelligent recognition technologies in agriculture, and providing technical support for agricultural production and resource management through digital platforms. This would provide a theoretical foundation for the innovation of digital agriculture and promote its sustainable development.

Assessment of health risks posed by heavy metal contamination of wild mushrooms and berries

BACKGROUND: One of the primary objectives in ensuring food security for the population is to reduce the mitigate the risk of chemical contamination in food. Wild mushrooms and berries have the potential to accumulate heavy metals from the environment, posing a threat to human health if consumed. AIM: To assess the risk of developing general toxic effects associated with the consumption of wild mushrooms and berries on health of the adults living in the Arkhangelsk region. MATERIAL AND METHODS: The amount and frequency of mushroom and berry consumption by the adult population of the Arkhangelsk region was assessed by a survey (n=445). Intake of heavy metals contaminating mushrooms and berries was calculated using four scenarios. Hazard Quotients (HQ) were used to characterize the risk of developing overall toxic effects due to exposure to mercury, arsenic, lead, and cadmium. Hazard Indexes (HI) for substances with unidirectional effects were employed to assess the risk of non-cancerous effects on critical organs and systems. Quantitative data were presented as medians (Me) with 95% confidence intervals (95% CI), and the 90th percentile (P90). RESULTS: In total, 82% and 70% of respondents were engaged in collection of mushrooms and wild berries, respectively. On average, they reported consuming 180 grams of fresh or frozen berries, 133 grams of berries with juice, and 50 grams of mushrooms in soup or boiled/fried form per week. The Hazard Quotients (HQ) for all heavy metals did not exceed 1.0. The Hazard Index (HI) calculated for the average consumption of mushrooms and berries, as well as the average heavy metal contamination of wild plants was below 1.0. However, at high levels of mushroom (P90 — 417 g/week) and berry (P90 — 900 g/week) consumption, along with high levels of heavy metal contamination, elevated risk of developing general toxic effects on the endocrine- (HI=2.27), cardiovascular- (HI=2.0), digestive- (HI=2.0), nervous- and immune systems (HI =1.81 for both) and kidneys (HI=1.25) were detected. Forest mushrooms and wild berries can be consumed without restriction at an average level of their contamination. However, in cases of heavy metal contamination of mushrooms and berries at the P90 level or above, it is not recommended to exceed daily consumption of 400 grams of forest mushrooms and 650 grams of wild berries. CONCLUSION: Consuming high amounts of wild mushrooms and berries at the upper limit of exposure to heavy metals, may lead to an elevated risk of developing toxic effects on endocrine, nervous, immune, cardiovascular, and digestive systems.

Rapid Risk Assessment of the Detection of Listeria Monocytogenes in Enoki Mushrooms

This rapid assessment has been requested to inform a decision by FSA Policy whether a temporary overarching risk management response to the commodity (enoki mushrooms) is necessary. Cultivated enoki mushrooms are deprived of light and grown in a CO2-rich environment. Commercially grown enoki mushrooms are grown in hard wood sawdust that is impregnated with the spores. The sawdust is usually good for four or five crops of mushrooms. The mushrooms are encouraged to grow long and thin by placing glass tubes over them. This produces a pale white mushroom with long, slender stems, up to five inches in length, and small caps. Whilst specific practices may vary, generally the initial growing environment is humid with a temperature between 22°C – 25°C, followed by lowering the temperature to 7°C – 12°C whilst maintaining humidity. It is this change in temperature that causes the mycelium to form fruiting bodies that grow into mature mushrooms which are harvested after about 90 days (Pereira et al., 2023). The species Flammulina filiformis, known and sold in different regions as “enoki”/“enokitake” Japan, “jingu” (China), “futu” (India), “paengi beoseot” (Korea), “nám kim cham” (Vietnam) also grows wild throughout a broad natural range on the rotting wood of a range of common trees, although wild-growing mushrooms appear quite different. The majority of enoki mushrooms on sale appear to be commercially cultivated and fresh rather than wild mushrooms. It is unknown if wild enoki mushrooms present a similar L. monocytogenes risk to cultivated mushrooms. Listeria spp., including L. monocytogenes, can be present in the soil (Linke et al., 2014), and are common in natural and urban environments (Orsi and Wiedmann, 2016). Contamination and proliferation is possible during cultivation but also during harvesting and packing (uncertainty).