SDF had a clean‐cut morphology whereas IDF had an indented microstructure with a wrinkled surface. After MW treatment, deep grooves and holes were observed. These variations in the IDF structure were more extensive. DF additions influenced water absorption, mixing tolerance index, dough development time, dough stability, resistance to extension, extensibility, energy of the dough and hardness, cohesiveness, springiness, chewiness, weight loss, specific volume, crust color difference of the bread in comparison with the properties of control samples significantly (p < .05). IDF had especially pronounced effects on the dough and bread characteristics. SDF enrichment provided more comparable results with the control bread than IDF. The originality of this work is to characterize isolated (100% purity) SDFs and IDFs, then discuss their effects on semi (dough) and final (bread) product quality.This study investigated the morphological and hydration properties of untreated and microwave (MW)‐treated isolate forms of soluble (SDF) and insoluble dietary fibers (IDF) obtained from grapes. Then, the rheological, textural, and other physical effects of the fibers (5% flour basis) were evaluated on bread quality. For this purpose, grape pomace was valorized as the juice extraction waste. MW significantly improved hydration properties of SDF and IDF by modifying their microstructures ( One hundred percent pure insoluble and soluble dietary fibers were obtained from grape waste by microwave treatment. The fiber properties were examined. These fibers were used separately for bread enrichment and the quality characteristics of the bread were determined. The best result was found in bread with microwave‐treated soluble dietary fibers. White flour is the main ingredient; thus, it affects the dough and bread's properties substantially. However, high consumption of white flour can lead to the development of insulin resistance, type II diabetes, and other disorders triggered by diabetes due to white flour's high glycemic index. Dietary fiber (DF) is a good alternative that reduces or eliminates the negative effects of white flour on health.In recent years, the role of DF in the prevention of diseases has been investigated. Ma et al.  reportedAnnual worldwide grape production was 78 million tons in 2020  have rapidly growing applications in food processing. MWs can alter the particle size, porosity, morphology, surface characteristics, hydration properties, molecular weight, thermal stability, crystallinity, monosaccharide composition, emulsion properties, and extraction yields of DFs  and then dried in a freeze dryer  under 0.1 mPa at −55°C for 48 h to a moisture content of 5.0 ± 0.4%. For preparation of the untreated samples, 100 g of the dried sample was ground for 30 s in a grinder . The ground samples were sieved through a 200‐mesh sieve (74 μm) (Endecotts). This sample was labeled as untreated GP. For preparation of the MW‐treated samples, 50 g of untreated GP was dispersed with deionized water (250 mL) in a beaker and treated in a microwave oven  at the parameters (800 watt/120 s) determined by preliminary experiments. For drying step, thoroughly mixed MW‐treated GP was poured into petri dishes and placed in a −80°C freezer with cooling to 20°C within 30 min. Samples frozen for 48 h were immediately placed in a freeze dryer and dried at −55°C for 72 h to a moisture content of 4.7%–0.5%. By following these steps, untreated and MW‐treated GPs were produced in an amount (300 g each) sufficient for analyses and enriched breads. Afterward, the SDFs and IDFs of the dried samples were isolated. SDF samples consisted of 99.67 ± 0.19% SDF and IDF comprised 99.55 ± 0.08% of IDF samples.2.22.2.1The SDF and IDF of the untreated and the MW‐treated GP were isolated and TDF, SDF, and IDF contents of the samples were determined by following the AOAC 991.43 procedure  all‐purpose wheat flour , 65% tap water, 3% compressed yeast (Saf), and 1.5% salt (Saxa). For the preparation of DF‐enriched bread samples, 5% DF fractions  were added to the dough prepared with 95% wheat flour. Since only the effect of SDF and IDF on dough and bread properties was investigated in the study, the amount of water was kept constant in all formulations.While preparing the dough samples, all ingredients were added and mixed in a Kitchen Aid mixer (5KSM125) for 10 min. The doughs were fermented in a proofer (Wachtel GmbH & Co.) at 35°C and 85% humidity for 70 min. Then, 250 g of dough was weighed into 20 cm × 10 cm × 7 cm pans and placed for 60 min in the proofer one more time. Dough fermentation parameters were used in proofing. In each run, three pieces of the dough were baked at 220°C for 25 min in a convection oven (Wachtel GmbH & Co.). After baking, all bread loaves were cooled for 60 min at room temperature. Then, the breads were subjected to measurements.2.2.3The moisture contents of the untreated GP and MW‐treated GP were determined using the thermal drying method in a drying oven  at 105°C for 12 h by following Wang et al.  procedurThe microstructures of the untreated and MW‐treated SDF and IDF samples were examined using a scanning electron microscope . Each sample was coated with gold with a sputter coater (Structure Probe) for 4 min before being scanned and photographed at 450× magnification. An accelerating potential of 5 kV was used during micrography.g for 20 min. The supernatant was poured into a tared beaker and put in an oven at 120°C to evaporate the moisture. The SI, WHC, SP, and SVF results of the SDF and IDF samples were calculated according to the Robertson et al. (The solubility index (SI), water‐holding capacity (WHC), swelling power (SP), and sediment volume fraction (SVF) of the SDF and IDF samples were measured at 25°C by using the centrifugal technique with some modifications  were reported from the extensograph test.The mixing profiles of the control and enriched doughs were determined using a farinograph  according to AACC Method 54–21 AACC, . Water a3/g), weight loss (%), and crust color difference (∆E*) of the breads were measured and the weight loss was calculated according to Equation Wei3) of breads were measured by seed (rape) displacement and the specific volumes were calculated using Equation SpeLₒ = 94.41, aₒ = −0.98, and bₒ = −0.35). The color difference (∆E*) for the crust was calculated using Equation (L is the lightness (brightness), a is redness: green to red, and b stands for yellowness: blue to yellow.Color values of the bread samples were detected in a Color Flex colorimeter (Hunter). The instrument was calibrated with standard tiles (black and white), and the color values of the white tile were taken as a reference  were obtained from Equation , cohesiveness, springiness, and chewiness (g) were measured from the midsection of the bread samples by using a Texture Analyzer  with a 36‐mm‐diameter cylindrical probe, 50% compression, and a test speed of 2.0 mm sEquation :(4)Che2.2.4n = 3). The significance level was p < 0.05.The statistical analysis was performed using SPSS 23 (IBM). All measurements were performed in triplicate. The control, U‐SDF, U‐IDF, MW‐SDF, and MW‐IDF groups were compared using an analysis of variance (ANOVA) by means of Tukey's HSD test. The results are presented as mean ± SD  MW oven, because MW‐treated samples were treated in a home‐type (2450 MHz) MW oven. In addition, AOAC 991.43 procedure was followed in DF analysis; thus, low‐molecular‐weight SDF content could not be measured in SDF samples.33.1The microstructures of the untreated and MW‐treated SDF and IDF are illustrated in Figure 3.2Hydration properties might affect the rheological, textural, and multiple other physical properties of the products. The SI, WHC, SP, and SVF values of the DF samples are summarized in Table SVF represents the insoluble fraction rate in the compound. For this reason, SVF values were found higher in IDFs, as expected. MW application reduced SVF scores in both DF groups. This reduction might be dependent on decreasing particle size due to the molecular collision generated by MW . These results could be explained as effects of the hydroxyl groups in the hydrocolloids, as these functional groups allow more water interaction through hydrogen bonding and might increase water absorption. The highest absorption was observed in the IDF‐enriched samples. Daou and Zhang . Rheological properties are presented as farinograph and extensograph measurements. The farinograph measurements present the parameters used to determine the kneading properties of the dough and provide information about the breading properties of the material used. The farinograph parameters of the control and the enriched doughs are summarized in Table nd Zhang  reached nd Zhang . A similnd Zhang  study onnd Zhang  in relatnd Zhang  found thRegarding the MW‐treated DF, the water absorption capacity rates of the dough with the MW‐treated DF increased. This finding is consistent with the WHC of the MW‐treated DFs. Absorption rates could be affected by the surface area expansion of the MW‐treated DFs due to particle size reduction. The developed holes and grooves in the microstructures of the MW‐treated DFs Figure  also supDF addition influences dough structure via different mechanisms: (1) DF dilutes and thereby weakens the gluten network; 2) DF molecules hinder the formation of bonds (steric effect) in the gluten network; (3) DF competes with gluten and starch molecules for water  of the dough fortified with DFs are presented in Table The enrichment of DF weakened the dough, which caused a decrease in its resistance to extension. The resistance to the extension values presented in Table Extensibility indicates dough handling characteristics. By adding water to the bread formula, glutenin and gliadin proteins in flour begin to hydrate, interact, and develop cross‐linked bonds that can hold the carbon dioxide produced by the yeast. This forms the extensibility of the dough. The influence of DF incorporation on dough extensibility was evident; according to the data in Table Significant energy reduction was observed after replacing flour Table . The cha3.43.4.1Certain quality characteristics of the bread samples are summarized in Table 3.4.22 captured  by entering between gluten molecules. This can weaken the dough and decrease the specific volume by reducing the amount of CO3.4.3The crust color difference values of the bread samples are presented in Table 3.5p < .05), as shown in Table Significant changes in textural properties were observed for all DF‐enriched breads p < .0, as showCohesiveness describes how well a product retains its form between the first and second chew. Springiness is the extent of recovery after the removal of force. Increased hardness is related to decreased cohesiveness and springiness. The results of this study are also consistent with this finding.As presented in Table Chewiness is like a summary of textural properties, as it expresses a combination of hardness, cohesiveness, and springiness; this property reflects the energy required to masticate food to a state ready for swallowing. SDF and IDF substitution markedly decreased bread chewiness. In fact, chewiness values are highly dependent on springiness; therefore, they were reduced (186.08–310.19 g) by decreasing springiness. The changes in springiness directly caused the changes in crumb elasticity. Similarly, the change in the chewiness of the breads, according to the changing trend of springiness, coincides with the Onyango et al.  study fo4In this study, MW treatment was utilized to modify the IDF and SDF of grapes, and the structural and functional properties of IDF and SDF and their effects on bread quality were investigated. The results of the SDF‐enriched samples were found to be closer to those of the control breads' values than those of the IDF‐enriched ones. MW treatment developed holes and grooves in the microstructures of DF and improved hydration properties compared to untreated DF. This improvement was also consistent with an increase in the SI, WHC, and SP of both DF fractions. IDF‐enriched breads had the highest farinograph measurements and the lowest scores in extensograph properties compared to the SDF‐enriched and control doughs. The weight loss, specific volume, crust color difference, cohesiveness, springiness, and chewiness of the IDF‐enriched breads were found to be the lowest. The addition of SDF provided farinographic, extensographic, textural, and other physical properties closer to the control than IDF fortification. MW treatment improved SDF's structure by helping it obtain water absorption rates, resistance to extension, energy, hardness, cohesiveness, and springiness scores closer to the control. It also modified IDF's properties to attain better resistance to extension, energy, hardness, and cohesiveness values in the IDF‐enriched breads compared to the control results. The importance of this study is as follows: (1) a 2‐min MW application changed the functional and technological properties of DFs; (2) in the enrichment of bread, it is recommended to use SDFs as a flour substitute at a certain rate (5% flour basis) rather than IDF; (3) MW treatment is more effective on SDFs than IDFs, and it is more effective in achieving results closer to control breads; (4) this study can serve as an example for food waste reduction, and these findings can provide a framework for further studies. For further research, the best bread formulations can be developed for SDF and IDF.Duygu Baskaya‐Sezer: Conceptualization (lead); data curation (lead); formal analysis (lead); investigation (lead); methodology (lead); resources (lead); software (lead); supervision (lead); validation (lead); visualization (lead); writing – original draft (lead); writing – review and editing (lead).This research did not receive any specific grant from funding agencies in the public, commercial, or not‐for‐profit sectors.The author has no relationship or conflict of interest with any person or company.The study does not need ethical approval. The material was not reproduced using other sources in the study.


Biological invasions threaten the functioning of ecosystems, biodiversity, and human well-being by degrading ecosystem services and eliciting massive economic costs. The European Union has historically been a hub for cultural development and global trade, and thus, has extensive opportunities for the introduction and spread of alien species. While reported costs of biological invasions to some member states have been recently assessed, ongoing knowledge gaps in taxonomic and spatio-temporal data suggest that these costs were considerably underestimated.InvaCost (v4.1)—the most comprehensive database on the costs of biological invasions—to assess the magnitude of this underestimation within the European Union via projections of current and future invasion costs. We used macroeconomic scaling and temporal modelling approaches to project available cost information over gaps in taxa, space, and time, thereby producing a more complete estimate for the European Union economy. We identified that only 259 out of 13,331 (~ 1%) known invasive alien species have reported costs in the European Union. Using a conservative subset of highly reliable, observed, country-level cost entries from 49 species , combined with the establishment data of alien species within European Union member states, we projected unreported cost data for all member states.We used the latest available cost data in Our corrected estimate of observed costs was potentially 501% higher (US$28.0 billion) than currently recorded. Using future projections of current estimates, we also identified a substantial increase in costs and costly species (US$148.2 billion) by 2040. We urge that cost reporting be improved to clarify the economic impacts of greatest concern, concomitant with coordinated international action to prevent and mitigate the impacts of invasive alien species in the European Union and globally.The online version contains supplementary material available at 10.1186/s12302-023-00750-3. Invasive alien species—those introduced into regions where they are not native and in which they cause negative impacts to nature and/or society—are among the main drivers of global biodiversity decline , 2, and Most assessments of the impacts of invasive alien species have relied on scoring systems, such as those proposed by Panetta  and subsInvaCost project and the structured and publicly available database it has produced [i) costs for many known, established invasive alien species that have not been assessed; (ii) cost information that is often inaccessible or otherwise not available publicly, or (iii) many cost estimates that do not fulfil quality criteria .Most recently, the produced –20, haveproduced , 22. Stuproduced ), regionproduced ), continproduced ), taxonoproduced ), or patproduced , 28) focIn Europe, the monetary costs incurred by invasive alien species from 1960 to 2020 were recently estimated to be US$140.2 billion , althougThe combined economic turnover of European Union member states makes it the third largest economy worldwide (data.worldbank.org), and Europe as a whole has been a historic centre of trade, human migration, and tourism. As such, both continental Europe and the European Union (with its current 27 member states) have been particularly vulnerable to biological invasions, despite recent transboundary legislation to tackle invasive alien species –33. The An absence of information on economic impacts risks downplaying the threats posed by invasive alien species (including costs and other impacts), because it underestimates the economic threat biological invasions represent and hinders national policies, governance, research, and educational efforts along with broad-scale multinational initiatives to combat the problem –38. ImprBecause cost information for many invasive alien species is missing, national and transnational estimates would benefit from a more complete accounting. Given that invasion history is one of the best predictors of eventual impacts , using ei) the projected amount of costs of invasive alien species will be considerably higher than those reported in the InvaCost (v4.1) database due to (ii) mismatches between known invaders and those with recorded costs [iii) expected increasing future trajectories of costs of invasions [Our analyses are therefore important methodological and applied advances towards improving spatial and temporal estimates of costs. We expected that  has standardised individual cost records to a common currency and year to account for variation and inflation: 2017 US$ . InvaCoanguages , and oveUS$ , Germany, Greece, Hungary, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, and Sweden. This filtering step identified 5,442 cost entries from the original global dataset .To ensure that we used cost data from only the 27 member states, we selected costs for every individual country currently in the European Union (excluding candidate countries), using the 'Official_country' column from the expandYearlyCosts function in the invacost package version 0.3–4 [InvaCost database [To obtain comparable costs of invasive alien species, we considered all costs lasting for a period of < 1 year as 'annual costs', and re-calculated (and thus ‘expanded’) costs covering multiple years on an annual basis by spreading the total cost across the time covered, using the on 0.3–4  in R veron 0.3–4 . Derivindatabase .InvaCost database to obtain a 'highly reliable, observed' dataset of only 'observed' costs using the 'Method_reliability', the ‘Method_reliability_refined’ and 'Implementation' columns reflecting (i) the perceived reliability of the type of publication and/or cost-estimation approach , and (ii) whether the cost was realised or empirically incurred ('observed') or whether it was based on predictions or costs expected over time or space . The resulting dataset of 'highly reliable' and 'observed' costs contained 5,592 entries. Further analyses focused on two descriptors: (i) type of cost: 'damage' refers to damage or losses incurred by invasion, and 'management' includes control-related expenditures ; (ii) impacted sector: the activity, societal, or market sector incurring the cost  quantified the ratio between recorded invasive alien species in InvaCost and established alien species nationally within European Union member states, and (ii) assessed the number of species of Union concern  2022/1203 of 12 July 2022 Amending Implementing Regulation (EU) 2016/1141 to Update the List of Invasive Alien Species of Union Concern. Available online: https://eur-lex.europa.eu/legal-content/EN/TXT/HTML/?uri=CELEX:32022R1203&from=EN) on invasive alien species recorded in InvaCost. For this, we assembled the most comprehensive dataset of established alien species in the European Union by combining the SInAS database of alien species occurrences [SInAS_AlienSpeciesDB_2.4.1 [To identify taxonomic gaps in the reporting of potentially costly invasive alien species, we , with the respective total number of invasive alien species recorded in InvaCost at the level of individual European Union member states. We also removed species not authenticated and thus, included only species-specific entries from the InvaCost database, thereby excluding any mixed or unspecified data.We applied a paired InvaCost database. We focused our analyses on country-level costs because they constitute most (92%) costs and are directly comparable. For full transparency, we also reported interpolated costs at the site level  where a given species’ impact had been estimated in at least one European Union member state, and we only used highly reliable observed, country-level costs reported in the We also tested for differences resulting from macroeconomic factors and to account for potential socio-economic differences, we analysed the following: GDP, population size, country area, and importance of the impacted sector (in %). We expected costs to increase with GDP and population size because these are typical macroeconomic predictors with broad data availability, and they were relevant socio-economic predictors for the costs of invasive alien species in previous studies , 24, 55.A/GDPB), and accounted for non-proportional effects using a fitted coefficient InvaCost-listed cost, s is species, c is country, h is type of cost , and j is impacted sector. M macroeconomic variables analysed. ci refers to each country i where species s has been observed, where we are trying to interpolate, whereas ck refers to the country where species k has been both observed and measured in InvaCost. To avoid unreasonable extrapolations beyond the data, we truncated scalar ratios by the maximum (or minimum) observed in the fitted model. The model allows for effects ranging from proportional scaling of costs with macroeconomics , using the deviation between predicted costs (Eq. 1) and costs observed in InvaCost for species–sector combinations where there were entries across multiple countries. We considered alternative models using all possible combinations of our four macroeconomic variables and calculated the Akaike's information criterion (AIC) for each model  and then without (n = 4872 entries) ‘extreme’ cost entries.To project the temporal dynamics of the monetary impacts at the European Union-level, we used the highly reliable and observed data subset. We then identified ‘extreme’ cost entries that can distort statistical analyses and violate model assumptions. These were any cost value exceeding the third quartile + 1.5 times the interquartile range for each year . Using tInvaCost (v4.1) changed over time, we applied a first-order autoregressive AR (1) process for the residuals [i) the values reported from 2018 to 2020 due to lag times in reporting of costs after they are incurred [ii) all years prior to 1980. We then modelled the annual total of this subset over time (using the 'Impact_year' of each cost entry as the year of occurrence) using both linear and quadratic robust regressions on the highly reliable, observed data via the lmrob function of the R robustbase package [n = 1,000. Ultimately, we projected the trend in total costs until 2040, by using the predict function in the forecast R package [To identify periods in which the variance of costs reported within esiduals . This enesiduals . This presiduals . Followiincurred , 62, and package , with ma package . Because package .InvaCost v4.1 amounted to US$129.9 billion , corresponding to €138.6 billion . Of these, US$7.3 billion  was highly reliable and observed, with US$752.9 million  dedicated to management costs. Both total and highly reliable, observed costs varied substantially across European Union member states  although this varied substantially by decade, falling from an initial US$19.17 million (€20.17 million) annually in the 1970s to US$0.5 million in the 1980s, before increasing to US$53.3 million (€56.1 million) in the 1990s and ultimately reaching US$461.1 million (€485.0 million) annually in the 2010s  established alien species on average recorded per European Union member state, ranging between 217 alien species established in Luxembourg and 3691 in France  totalled US$4.7 billion . On this conservative basis of highly reliable, country-level costs, the different projections , from which country-level costs , followed by Malta , Czech Republic , and Denmark . In terms of raw costs, the highest increase was in Czech Republic , followed by Germany , France , the Netherlands and Poland both with US$2.0 billion (€2.1 billion). Added costs for all other countries were < US$2 billion (€2.1 billion). We provide a detailed breakdown of projected country-level costs in Fig. InvaCost database). Finally, scenarios considering the missing costs of the two main types of costs resulted in projections increasing management costs by US$8,786.76 million (3933%), and by US$14,400.38 million (325%) for damage costs , we identified substantial increases with both linear and quadratic robust regressions. This increase held even without extreme values, albeit reducing the US$7.3 billion (€7.7 billion) in highly reliable, observed costs to US$420.6 million (€442.4 million), with US$116.6 million  associated with management expenditure. Linear and quadratic robust regressions estimated similar slopes, but including extreme values estimated a larger increase in projected trends. Projected costs for 2040 ranged from US$21.1 billion (€22.2 billion) to US$30.6 billion (€32.2 billion) excluding extreme values; and from US$42.2 billion (€44.4 billion) to US$148.2 billion (€155.9 billion) including extreme values. This was also reflected in projected increases in the total number of invasive alien species  that has already been incurred, we have shown that after accounting for taxonomic and geographic gaps in cost reporting, the magnitude of invasion costs was at least 1.5 to 6 times higher, even while using a conservative subset of the available data. We expect these costs to rise further towards 2040 in accordance with the temporal increase in numbers of alien species . In addiEuropean Alien Species Database [Economic costs are available for just 1% of alien species recorded as established across the European Union. For the 88 invasive alien species currently identified as being of Union concern, a much higher proportion of 54.5% had reported costs . This absence of data is concerning given that ~ 14,000 alien taxa are listed in the Database  for EuroDatabase . This din = 3080 entries) with costs inferred only at the level of specific sites. We found that growth due to the scaling of missing costs was particularly burdensome to the public and social-welfare sector, followed by sectors encompassed by the circular economy initiatives and the European Green Deal . This information is often unavailable in underlying studies, but would constitute a major step forward in understanding management decisions for future work . HoweverInvaCost, a living database that has grown considerably in only a few years . Simplifying the process of adding information to such databases would facilitate use and uptake. This should include increasing outreach to industry groups, health insurance sectors, and others where a lack of communication among those who are affected by damage or loss, those who assess costs, and those who synthesise costs results in currently recorded but inaccessible data. Such efforts would provide a more complete and accurate picture of the total costs and the full economic impacts of biological invasions. While technical updates to database accessibility are complex and ongoing, our scaled projection approach provides a more complete estimate of costs and a major advance in the interpolation of invasion costs from limited data. Unlike previous attempts to investigate the temporal dynamics of the damage costs of invasive alien species [Although not all alien species do or will cause monetisable damage or elicit management expenditure , th species , 75, theOryctolagus cuniculus in Germany in the 1970s; [InvaCost. The projected increase in the number of invasive alien species reflects a robust predicted rise identified in invasion rates and an increasing awareness of their impacts [Early cost reporting for invasive alien species was particularly sporadic , with c impacts .The well-characterised effect of reporting lags in the costs of invasive alien species , 20, 80 Ambrosia artemisiifolia and can potentially be explained by the large proportion of agricultural land invaded . Out of all the European Union member states, Romania and Hungary have notably higher reported costs resulting from invasive species at the country level. These high costs are driven by estimated damages caused by the common ragweed ctively; ). This iBecause we estimated potential costs caused by invasive species for European Union member states where monetary impacts were unknown, it is important to emphasise the inherent uncertainty involved. Consequently, projected costs, whether over space or time, should be considered with caution. Nevertheless, we also note that these estimations were based only on a conservative subset of the richest country-level data, while considering that for invasive alien species from which models were fit, the costs were still fragmented and underestimated, thereby highlighting the importance of the costs involved. An additional source of uncertainty lies in lack of knowledge about the efficiency of management costs. Information on whether management expenditure has turned out to be efficient and to what degree, in relation to their objectives, can be essential for building a more comprehensive understanding of the true costs associated with biological invasions.InvaCost database. Examples could include, for instance, an increase in biological production in semi-arid ecosystems, the restoration of industrial or contaminated areas where native species cannot survive, with benefits from the acceleration of succession and soil formation or reductions in nutrient leakage and water basin eutrophication [Buddleja davidii might provide certain local economic benefits [Our analyses do not consider any positive effects of invasive alien species, which are generally not captured in the hication , 85, or hication . Alternabenefits , 88, altbenefits , and nori) improved biosecurity protocols; (ii) well-coordinated, large-scale management; (iii) targeted research to improve pre- and post-invasion mitigation; and (iv) horizon scanning to include a consideration of costs [The number of invasive alien species and resultant economic costs will likely increase in the future, even if projections remain uncertain , 90, dueof costs . Well-coof costs , 93, 94.Cost assessments play an important role in improving management efficiency and effectiveness by promoting early, focused, and evidence-based management interventions towards alien species, ultimately leading to long-term cost savings, relieving the burden on taxpayers, and enabling governments to allocate their financial resources more effectively. This is further substantiated because prior work has emphasised the ability of early investments  to lower impacts of biological invasions in the long term , 93. HowAdditional file 1: Description of the sectors considered in the InvaCost database. Additional file 2: Site-level costs recorded in InvaCost for the European Union member states (a), by type of cost (b) and impacted sector (c) with and without high-leverage points and the respective projected additional cost and percent increase. Additional file 3: Detailed breakdown of the considered models included in the model averaging. Table 1: Country-level analyses. Values of the fitted parameters for each predictor combination and model performance estimated using Akaike’s information criterion (AIC). We calculated Akaike’s weights for model averaging following Burnham & Anderson [Table 2: Country-level analyses without high-leverage points. Values of the fitted parameters for each predictor combination and model performance estimated using Akaike’s information criterion (AIC). Model outputs for each combination of predictors. We calculated Akaike’s weights for model averaging following Burnham & Anderson [Table 3: Site-level analyses (A) with all data and (B) without high-leverage points. Values of the fitted parameters for each predictor combination and model performance estimated using Akaike’s information criterion (AIC). Model outputs for each combination of predictors. We calculated Akaike’s weights for model averaging following Burnham & Anderson [Additional file 4: Invasion costs  for European Union member states recorded in InvaCost v4.1, in 2017 US$ billion. Additional file 5: Comparison of the number of established alien species in the European Union and alien species in InvaCost v4.1. Additional file 6. Established alien species of Union concern recorded among the established alien species in the European Union with the first European Union member state they were recorded in and the year of first record [Additional file 7: Breakdown of interpolated costs excluding high-leverage points. Additional file 8: Country-level costs recorded in InvaCost with and without high-leverage points and the respective projected additional cost and percent increase. Additional file 9: Country-level costs recorded in InvaCost by type of cost (a) and impacted sector (b) with and without extreme values and the respective projected additional cost and percent increase. Additional file 10: Temporal projection of total annual management costs (a), recorded species per year with management costs (b), and number of reported references on management expenditure in InvaCost v4.1 per year (c) using linear  and quadratic  robust regressions and their respective confidence intervals. Solid dots and lines represent trends without the inclusion of extreme values. Open circles and dashed lines represent the respective trends when including all data . Grey dots present data for the period 1960–1979 that are not considered in the projection due to an absence of repeated measures of individual cost entries.Anderson . Table 2Anderson . Table 3Anderson . Additiot record . Additio


Acanthamoeba polyphaga mimivirus, so called because of its “mimicking microbe”, was discovered in 2003 and was the founding member of the first family of giant viruses isolated from amoeba. These giant viruses, present in various environments, have opened up a previously unexplored field of virology. Since 2003, many other giant viruses have been isolated, founding new families and taxonomical groups. These include a new giant virus which was isolated in 2015, the result of the first co-culture on Vermamoeba vermiformis. This new giant virus was named “Faustovirus”. Its closest known relative at that time was African Swine Fever Virus. Pacmanvirus and Kaumoebavirus were subsequently discovered, exhibiting phylogenetic clustering with the two previous viruses and forming a new group with a putative common ancestor. In this study, we aimed to summarise the main features of the members of this group of giant viruses, including Abalone Asfarvirus, African Swine Fever Virus, Faustovirus, Pacmanvirus, and Kaumoebavirus. Nucleocytoviricota [Phycodnaviridae, Asfarviridae, Poxviridae, Iridoviridae, and Ascoviridae. Their broad host range includes algae for Phycodnaviruses [Heterocapsa circularisquama [Vermamoeba vermiformis as a novel cell support for the co-culture of giant viruses made it possible to isolate Faustovirus, a new virus closely related to ASFV [Kaumoebavirus, a phylogenetically close relative of Faustovirus, was isolated from wastewater collected in Jeddah, Saudi Arabia [Pacmanvirus strain A23 was isolated from sewage collected in Oran, Algeria by co-culture with Acanthamoeba castellanii. This virus, which has very different morphological features, phylogenetically clusters with the pre-cited extended clade of Asfarviridae [The serendipitous discovery of giant viruses of amoebas broadened the field of virology in a sudden and spectacular way , due to viricota . This phviricota . Historiaviruses , mainly aviruses , insectsaviruses  and Entoaviruses . Africanaviruses , and cauaviruses . In 2009risquama , reveali to ASFV . In 2016i Arabia . In 2017rviridae .Kaumoebavirus [In this study, we aimed to describe the main features of the members of the extended clade of Asfarviruses, including the close relatives isolated by co-culture of protist amoebae and discovered more recently such as Faustoviruses , Kaumoebebavirus , Pacmanvebavirus , Abaloneebavirus , and MetAsfarviridae (African Swine Fever and relatives) in which the Asfivirus is the only genus recognised and ASFV is the only member [This virus has long been classified as an Iridovirus ,16. Howey member . ASFV way member . In Easty member ,20,21,22y member ,24 whereOrnithodoros that also constitute a viral reservoir [ASFV is the causative agent of African swine fever, a haemorrhagic syndrome with a mortality rate close to 100% in its acute forms in pigs, warthogs, and wild boars of all age groups  in Europeservoir ,29,30. Ieservoir . In its eservoir ,33. ASFVeservoir ,34,35 aneservoir ,37,38,39eservoir  against eservoir ,42, whiceservoir ,44. ASFVeservoir ,46.The icosahedral virions are about 250 nm in diameter ,48 and eThe ASFV genome consists of a linear double-stranded DNA molecule with a length of approximately 170–194 kb predicting to encode 150–167 Open Reading Frames (ORFs), depending on the isolate. The GC content of the ASFV genome is around 39% ,51,52,58ASFV multiplies in the monocytes and macrophages in the blood and bone marrow of pigs. Viral replication has also been observed in endothelial cells, hepatocytes, renal tubular epithelial cells ,60,61,62Phacochoerus africanus) and soft ticks belonging to the genus Ornithodoros. Juvenile warthogs infected by soft ticks are viraemic, but do not develop any disease and constitute the source of infection for naive ticks taking a blood meal. In Europe, transmission of the virus is usually directly from a sick pig to a healthy pig or indirectly via a mechanical vector such as soiled premises, vehicles, instruments, or clothing [Asfarviruses represent the only known DNA virus transmitted by arthropods . In Africlothing ,37, but clothing . Severalclothing ,71. Tranclothing ,73. Theyclothing .The virus then passes into the retropharyngeal lymph nodes, whence it disseminates into the blood. The incubation period of the disease may vary between 4 and 19 days . ClinicaThe main manifestations of the infection are thrombocytopaenia, leukopaenia, lymphopaenia,  and haemAcanthamoeba was used for giant virus isolation. In 2015, the implementation of a high throughput culture strategy by diversifying the cell supports made it possible to isolate a new giant virus: Faustovirus E12 [Vermamoeba vermiformis, which is reported to be the most common free-living protist in human environments [Historically, only the genus ronments ,77,78. Tronments  and 16 oronments ,81,82,83ronments .Faustoviruses virions harbour icosahedral symmetry and have a diameter of about 250 nm, as determined by cryo-electron microscopy ,85. FausAsfarviridae members (31–45%) [By the start of 2023, 18 complete genomes of Faustoviruses had been published. Genome lengths range from 455,803 bp (for Faustovirus E3) to 491,024 bp (for Faustovirus E9) ,88. The (31–45%) ,89. Thei(31–45%) . The Fau(31–45%) ,86,90 seGenome-wide alignments of Faustoviruses revealed a high level of co-linearity, but an increased indel accumulation and local rearrangements have been noted more frequently in the Major Capsid Protein gene region and the extremities of the genomes, revealing a faster rate of evolution. In addition, most Faustoviruses have terminal inverted repeats (TIRs) at both ends of their genome , so the Phylogenetic analyses based on the core genes, especially the DNA polymerase B, show that Faustovirus is a distant relative of Asfarviridae . A more V. vermiformis lasts 18–20 h, which is slightly shorter than that of Mimivirus (14 h) [The entire Faustovirus replication cycle in s (14 h) . At the s (14 h) .Vermamoeba vermiformis is, to date, the only host of Faustovirus [stovirus . This histovirus . FaustovV. vermiformis. The strain Kaumoebavirus KV-Sc [Kaumoebaviruses are the second giant viruses isolated by co-culture on us KV-Sc  was the us KV-Sc .Kaumoebavirus virions are icosahedral particles of ∼250–260 nm in diameter, devoid of fibrils.The Kaumoebavirus genome consists of a 350 kbp to 362.6 kbp double-stranded DNA linear molecule, which is shorter than those of Faustoviruses, with a GC content ranging from 43.1% to 43.7% ,95. TheyAsfarviridae members and Faustoviruses, A/T rich motifs are abundant, widely distributed upstream to start codons, especially upstream to the core genes of Asfarviruses, Faustoviruses, and Kaumoebavirus, and play an important role as promoter sequences [Asfarviridae. Interestingly, the region predicted to encode the MCP shows intron differences between the Sc and LCC10 strains. However, and as is the case in Faustovirus, this region contains ORFs encoding a GIY-YIG endonuclease. This specific region is in the middle of the genome and seems to have a higher rate of evolution.The genomes of two strains both encode for 465 genes and are collinear for the main part of their length, albeit with a total of 180 ORFs detected as lost or gained in one or other genome. A total of 59% of Kaumoebavirus KV-Sc proteins are homologous with other viral proteins, including 43% with Faustovirus and 23% with Asfarviruses. It is noteworthy that 67–76% of their gene content are ORFans. Although Faustoviruses are the closest relatives, genome co-linearity is not conserved with Kaumoebavirus KV-Sc ,95. As fequences . NotablyVermamoeba vermiformis through phagocytosis. At three hours post-infection, virions are often visualised as clusters of two to four particles in vacuoles. The “eclipse” phase occurs at four hours post-infection, followed by the formation of viral factories at six hours post-infection. New Kaumoebavirus particles fill viral factories until 16 h post-infection and are released at 20 h post-infection through cell lysis. The nuclear morphology of the host cells is not modified during the replication cycle of Kaumoebavirus, in contrast to what happens with Faustovirus [The replication cycle of KV-Sc lasts between 16 and 20 hours. Kaumoebavirus particles enter into stovirus .A. castellanii [Acanthamoeba castellanii [Pacmanvirus lupus, isolated from a Siberian Canis lupus intestine collected in permafrost  [Pacmanvirus A23 was isolated from a water sample collected from a well on a roadside in Algeria (North Africa) , by co-ctellanii ,98. The tellanii . A secontellanii . A third0 years) .Pacmanvirus virions are icosahedral in shape with an overall external structure close to that of Faustovirus, of about 250 nm in diameter. The major capsid protein of Pacmanvirus is double jelly-roll fold, as confirmed by three-dimensional cryo-EM reconstruction, and it has an inner membrane (IMV) but no distinct inner capsid shell . The capThe complete genome of Pacmanvirus A23 is linear shaped and reaches 395,405 bp in length, which is shorter than Faustoviruses and longer than ASFV and Kaumoebaviruses. It encodes for 465 genes with a GC content of 33.6%, with the particularity of not having large, repeated regions, as is observed for Kaumoebaviruses and ASFV, but not for Faustoviruses. Of these 465 genes, about half (244 ORFs) encode for proteins with no hits in the nr database. Only one-third encode for proteins with a functional annotation. Moreover, a single tRNA (Ile-tRNA) also detected in six Asfarvirus MAGs, is present in the genomes of Pacmanvirus A23 and S19 . The PacAcanthamoeba castellanii for strain A23. The cycle starts with phagocytosis of the virions. Pacmanvirus seems to be able to escape the phagosomal step, as observed by a capsid opening that does not occur immediately. A few minutes after phagocytosis, once in the host’s cytoplasm, some virions migrate close to the mitochondria, but without membrane fusion. Although empty capsids have been detected in the cytoplasm, DNA release from the capsid has not been observed to date [The replication cycle of Pacmanvirus A23 has been exhaustively studied by transmission electron microscopy . Pacmanv to date . Early v to date .Haliotis spp. [Haliotis diversicolor, and the virus has a preferential tropism for the gill [In 2020, a novel Asfarvirus-like virus, provisionally called Abalone asfar-like virus (AbALV), was identified as one causative agent of abalone amyotrophia, responsible for mass mortality in this gastropod mollusc . Other vtis spp. . The inithe gill . This fiPhytophtora parasitica) [Elysia marginata [bona fide reflection of an Elysia spp. infection. Nevertheless, it could be very interesting to find expanded Asfarviridae sequences in gastropods, as its the case in abalone species for AbALV. In 2021, a study by Karki et al. [The expansion of next generation sequencing and metagenomic sequencing has resulted in numerous sequences becoming available in public databases. This has two major consequences: numerous mis-annotations using automatic annotation ,104 partasitica) . This viarginata , an aquai et al.  identifiAs the latest pan-metagenomic analyses have shown, the number of shared proteins in this group is reduced to about ten among more than 7000 with an extreme diversity of genome size and genomic content . PhylogeAsfarviridae family. The recent isolation of these new giant viruses suggest a great diversity, which remains far from being completely elucidated. The intense economic interest in ASFV disease and the study of a vaccine candidate for pigs associated with the isolation and detection of these viruses in amoebae, gastropods and different marine metagenomes, illustrates the great advances made in recent years in Asfarviridae. Structural studies describe a complex capsid organisation for this group, expanding our knowledge of Nucleocytoviricota emphasising the separation between two sub-groups ASFV and abalone virus on the one hand and viruses from amoeba on the other.Historically, ASFV was the only member of this


Plants can be infected with multiple viruses. High-throughput sequencing tools have enabled numerous discoveries of multi-strain infections, when more than one viral strain or divergent genomic variant infects a single plant. Here, we investigated small interfering RNAs (siRNAs) in a single strawberry plant co-infected with several strains of strawberry mottle virus (SMoV), strawberry crinkle virus (SCV) and strawberry virus 1 (StrV-1). A range of plants infected with subsets of the initial viral species and strains that were obtained by aphid-mediated transmission were also evaluated. Using high-throughput sequencing, we characterized the small RNA fractions associated with different genotypes of these three viruses and determined small RNA hotspot regions in viral genomes. A comparison of virus-specific siRNA (vsiRNA) abundance with relative viral concentrations did not reveal any consistent agreement. Strawberry mottle virus strains exhibiting considerable variations in concentrations were found to be associated with comparable quantities of vsiRNAs. Additionally, by estimating the specificity of siRNAs to different viral strains, we observed that a substantial pool of vsiRNAs could target all SMoV strains, while strain-specific vsiRNAs predominantly targeted rhabdoviruses, SCV and StrV-1. This highlights the intricate nature and potential interference of the antiviral response within a single infected plant when multiple viruses are present. RNA silencing is a conserved mechanism invoked not only to counteract invading viral and viroid species, but also to regulate gene expression . Double-Analyzing the vsiRNA pattern of virus-infected plants has shown that specific genomic regions give rise to greater numbers of vsiRNAs, termed RNA silencing ‘hotspots’ .In practice, the characteristics of virus-specific siRNAs can be used for developing induced virus resistance ,8,9, witClosteroviridae) and potato virus Y (Potyviridae) [The use of high-throughput sequencing has revealed that mixed viral infections are common in nature ,13,14,15viridae) ,25,26,27viridae) , but notviridae) ,29.The mechanisms underlying cross-protection are not fully understood. Remarkably, while some studies suggest that RNA silencing is not involved in the process ,30, otheSecoviridae), strawberry crinkle virus (SCV—Rhabdoviridae) and strawberry virus 1 (StrV-1—Rhabdoviridae) [Over thirty different viruses are able to infect strawberry ,36,37,38viridae) ,43,44. Aviridae) ,46,47; hviridae) .In the current study, we focused on the vsiRNA analysis of strawberry plants infected with several viruses in different combinations, characterized the specificity of vsiRNAs against individual viral strains, and evaluated whether the relative viral concentrations correspond to the levels of vsiRNA.Fragaria ananassa cv Cacanska rana CRM plant (CRM is an acronym for Cacanska rana mother) was infected with a mixture of three viral species: SMoV, SCV and StrV-1. In total, eleven distinct viral RNAs were detected: three for each genomic component of SMoV, two strains of SCV and three strains of StrV-1 [The original f StrV-1 ,43. The Stolons from the original plant were used for vegetative propagation. Three randomly selected daughter plants were used for small RNA sequencing library preparation. From each plant, three to four libraries were prepared from different leaves, resulting in eleven datasets (1B—four datasets (1B_1—1B_4), 2A—three datasets (2A_2—2A_4), and 3K—four datasets (3K_1—3K_4)). The samples were verified in parallel with RT-qPCR  and RNA1C ; t (7.1) = −0.52, p = 0.62. Correspondingly, no significant difference was observed for RNA2A  and RNA2C ; t (7.7) = 0.17, p = 0.89. Thus, future studies of SMoV strain variability should further examine these findings. Taking into account the considerable difference in the titers of RNA1B and RNA1C, it would be problematic to detect RNA1B without prior knowledge of its existence. Nevertheless, as the actual variability of SMoV strains was uncovered only a few years ago, reliable tools for the identification of the full spectrum of SMoV strains may still be lacking. The existing SMoV detection system relies on the highly conserved stretch within the 3′UTR of both RNA1 and RNA2, which is a superb region from a high conservation perspective, but a poor choice for strain discrimination.Therefore, we hypothesized that the RNA1ed reads B betweenGATGTTATCATCCGT-3′ aligned against the StrV-1 genomic strand at its 5′ end (positions 2–28). An analysis of potential secondary structures of the sequence showed only a weak structure with a four-base-long loop (underlined). The genesis of vsiRNA involves the production of mainly 21 nt and 22 nt long products, and the presence of 24 nt vsiRNAs has been reported only for some DNA and RNA viruses [Considering the observed 27 nt hotspot in the StrV-1_B strain, we analyzed the small RNA population in plants infected with either StrV-1_A or StrV-1_B, or a combination thereof . On aver viruses ,57. Duri viruses ; hence,  viruses , which wFurthermore, comparing the number of vsiRNAs per virus in the different samples showed that there were 14,000 mapped reads on average in a single StrV-1 infection, while the corresponding average when other viruses were present was much lower, at 3500 . SMoV enOur data show that plants infected with more than one viral strain produces strain-specific vsiRNAs against each viral RNA. For SCV, the vsiRNA hotspot was found at the 3′ terminus of the genomic RNA, whereas no universal hotspot was found for StrV-1. Furthermore, although both the SCV and StrV-1 vsiRNAs were quite specific, a large fraction of SMoV vsiRNAs matched more than one strain due to its long conserved 3′UTR regions. It seems that in the development of a plant protection method based on exogenous siRNAs against SMoV, 3′ UTRs might be a good target. However, there was no consistent agreement between the relative levels of viral RNA and the corresponding levels of vsiRNAs. Despite the very low abundance of SMoV RNA1B in Fv-SMoV_BC_AC-1 and -2 plants, the levels of RNA1B-specific vsiRNAs were not significantly lower than those of vsiRNAs targeting the other strain or other viruses. When loaded into the RISC, vsiRNAs in multiple infections can target related viral strains, which is why it is very difficult to find a direct correlation between the concentration of different viral strains and the number of vsiRNAs. The presence of viral silencing suppressors capable of interfering with antiviral silencing in different steps of RNAi can further alter this complex pattern, which should be investigated in future studies.F. ananassa, cultivar Cacanska rana, served as a plant source for stolon-mediated propagation  and mirPremier microRNA Isolation Kit , respectively, following manufacturers’ recommendations. The quantification and quality control of the RNA extracts were performed using a Nanodrop 1000 UV-Vis spectrophotometer and Qubit HS RNA and IQ assays .Total RNA  was reverse-transcribed to cDNA using the Maxima H Minus First Strand cDNA Synthesis Kit with dsDNase in 20 µL reactions following the manufacturers’ recommendations. The cDNA was then diluted to a ratio of 1:10 with Milli-Q-grade water and subjected to qPCR assays.The RT-qPCR assays were conducted using a CFX96 real-time PCR detection system . The 10 µL reaction was prepared from 5 µL of tenfold-diluted cDNA, 0.25 µL forward and reverse primers , 2.75 μL of nuclease-free water and 2 µL of 5x HOT FIREPol EvaGreen qPCR Mix Plus . The reaction conditions were 95 °C for 12 min, followed by 40 cycles of 95 °C for 10 s, 60 °C for 20 s and 72 °C for 20 s. The dissociation curve analysis was performed by ramping from 65 °C to 95 °C (with increments of 0.5 °C for 5 s) to verify the specificity of primer amplification and the presence of potential primer dimers based on a single peak. No template, positive, and if necessary, no reverse transcriptase controls were included to check for potential cross-contamination and the presence of genomic DNA. The amplification efficiency E,  was asseref − Cq virus), where efficiency is the experimentally calculated efficiency of the corresponding primers  and Cqvirus is the Cq value of the viral strain.Relative viral concentrations were calculated using the formula efficiency ^  and R software 4.3.0 .The plants and respective sequencing libraries are listed in Specifically, the minimum identity was set to 100%, and the reads matching more than one reference sequence were mapped either to all or none of them, depending on the required result. Assuming the presence of more than one strain in the data, we set mapping conditions that were rather stringent, without allowed mismatches. The mapping settings allowed us to establish rules for a read matching more than one position (reference): (a) discard the read, (b) map it to all matching positions or (c) map it to only one position through a random algorithm. In the current study, we used the first two options. When such reads were mapped to all references, they were referred to as ‘mapped reads’; when such reads were discarded, thus retaining only reference-unique reads, they were referred to as ‘strain-specific’ .The read counts were analyzed using R software 4.3.0  and Expl


In this study, the effects of enzymatic modification using cellulase/xylanase on the composition and structural and functional properties of ginseng insoluble dietary fiber (G-IDF) were evaluated. Fourier transform infrared spectroscopy and scanning electron microcopy showed that enzymatic extraction treatment caused obvious structural alterations in ginseng-modified (G-MIDF) samples, which exhibited more porous and completely wrinkled surfaces. Comparing the peak morphology of G-MIDF with untreated IDF using X-ray diffractometry, the G-MIDF sample exhibited split peaks at a 2θ angle of 23.71°, along with the emergence of sharp peaks at 28.02°, 31.78°, and 35.07°. Thermo-gravimetric analysis showed that G-MIDF exhibited a specified range of pyrolysis temperature and is suitable for food applications involving processing at temperatures below 300 °C. Overall, it was evident from rheograms that both G-IDF and G-MIDF exhibited a resemblance with respect to viscosity changes as a function of the shear rate. Enzymatic treatment led to significant ( Panax ginseng C.A. Meyer) is a perennial plant that belongs to the Araliaceae family and is native to the mountainous regions of eastern Asia, including China, Korea, and Siberia [Ginseng  are complex carbohydrate polymers composed of at least ten monomeric units, resistant to hydrolysis by the body’s own enzymes in the small intestine. Insoluble fiber (IDF) is part of the composition of the DFs, and IDF has been reported to comprise several functional groups including phenolics, carboxylic acids, ketones and aldehydes . IDF hasGinseng IDF is a non-starch polysaccharide that is made up of cellulose, hemicellulose, and lignin. Ginseng IDF has a number of health benefits. Ginseng IDF can help to regulate bowel movements and promote the growth of beneficial bacteria in the gut. This can help to improve digestion and reduce the risk of constipation, diarrhea, and other digestive problems . GinsengThe inclusion of insoluble dietary fibers (IDF) in food products can lead to quality problems in terms of flavor, color, and texture. To address these issues, various methods for modifying fibers like enzymatic, fermentation, as well as chemical processes have been reported in the published literature. Chemical extraction methods, including acidic, alkaline, chelating, and oxidant extractions are commonly utilized to obtain dietary fibers (DFs), with the method selection being dependent on the desired polysaccharide . AdditioFrom a local supermarket in Jilin, China, dried ginseng roots were purchased. The ginseng roots were subject to cultivation for a period of 5 years, and procurement was carried out in the season of November 2022. The preparation of ginseng residue was carried out following the method of Jiang et al. . The polIDF preparation was performed using ginseng residue, as described by Kurek et al. , with slw/v = 1:20) for pH value adjustment to 4.9. Then, 0.25% cellulase/xylanase solution : xylanase (U) = 1:1) was taken and subjected to rigorous mixing with PBS at a temperature of 60 °C in an oscillating water bath for hydrolyzation for 1.5 h. For the inactivation of the enzymes, the mixture solution was subjected to boiling for a period of 15 min. The solution was subjected to centrifugation for a time period of 10 min at the centrifugation rate of 4000× g. The, the enzymatically modified IDF (MIDF) was utilized to extract the residue and then subjected to washing and drying. The modification of ginseng IDF was carried out based on a modified method described by Ma et al. . GinsengThe morphological attributes and microstructural properties of IDF and its MIDF were evaluated by means of SEM . The samples were subjected to slicing into rectangular shapes by keeping them on the horizontal plane. The sample was mounted on a specimen holder, followed by gold sputter-coating. The microstructural attributes were analyzed using SEM . Capturing of the SEM imagery was performed at magnification levels of 1000× and 2000×.−1 at 4 cm−1 resolution at <10 s scan speed. The FT-IR attenuated total reflection (ATR) mode was employed to obtain FT-IR spectra. An FT-IR spectrophotometer  was employed for analyzing the organic functional groups present in ginseng IDF as well as MIDF samples. The FT-IR spectra were recorded at the wavelength range of 400–4000 cmThe XRD profiles of IDF and MIDF samples were determined by means of a diffractometer . The equipment specifications for analysis were as follows: diffraction angle (2θ) falling in the range of 4–40°, as well as copper Kα radiation .A thermogravimetric analyzer  was utilized for the evaluation of ginseng IDF samples (5 mg). The temperature was in the range of 30–600 °C, whereas the heating rate was set at a rate of 20 °C/min under nitrogen atmosphere.w/v on basis of dry weight by employing a magnetic stirrer at ambient temperature for 10 min. The heating temperature of the solution was elevated to 80 °C, aided by constant stirring and temperature maintenance for a time period of 15 min. After reaching the heating temperature, the solution mixture was subjected to cooling with constant stirring to an ambient temperature. Finally, the solutions were stored at a temperature of 4 °C for 24 h.The dissolution of IDF samples in sodium phosphate buffer  was carried out for the preparation of 1% solutions mixture in terms of −1 at an operational temperature of 25 °C. With a corresponding rising trend of shear rate ranging from 0.10 to 1000 s−1, the measurement of sample viscosities was performed. All samples were assessed thrice. IDF and MIDF were evaluated in terms of apparent viscosity (η), consistency coefficient (K), and flow behavior index (n). The experimental data were subjected to fitting in accordance with the power law model equation given below:−1) was denoted by η, consistency coefficient (Pa·s n) by K and the flow behavior index by n.Rheological measurements were carried out for SDF solutions by employing a rheometer . Steel cone geometry with a diameter of 60 mm and gap of 56 µm was utilized to perform the analysis. Continuous shear test was applied with a shear rate ranging from 0.10 to 1000 s1 denotes to sample total dry weight and W2 is the sample total weight after water has been filtered out. The water holding, (WHC), oil holding (OHC), and water swelling capacities (WSC) were evaluated in accordance with the method reported by Meng et al. . In brieg for a time interval of 10 min. Following centrifugation the centrifuge tube was inverted in order to discard excessive amount of soybean oil. The OHC was then quantified as per the equation given below: 1 and the sample’s total weight calculated after discarding excess oil is denoted by W2. A sample accurately weighed to a specified amount of 0.5 g was added to the centrifuge tube (10 mL) and subjected to mixing with distilled water (5 mL). Then, the reaction mixture was added for incubation for 24 h at a temperature of 37 °C. Then, WSC was assessed as per the equation given below: 1, V2 is indicative of hydrated sample volume (mL), and DF weight prior to hydration (g) is represented by W. OHC assessment of IDF and MIDF was performed. In brief, a sample accurately weighed to 0.5 g was subjected to mixing with soybean oil in a centrifuge tube followed by incubation for 1 h at a temperature of 37 °C. Then, the sample was subjected to a process of centrifugation at 5000× The method reported by Luo et al.  was emplMeasurement of the BAC was performed in accordance with the method of Luo et al. . In brieFor calculation of the CAC, the reported method by Luo et al.  was emplg. Determination of the final glucose concentration of the collected supernatant was performed by means of a glucose (oxidaseperoxidase) assay kit . Then, the GAC was quantified and expressed in terms of mg/g. In brief, an IDF sample accurately weighed to 0.5 g was subjected to mixing with 50 mL glucose solutions . After being subjected to rigorous stirring in a continuous manner for a time interval of 2 h at a temperature of 37 °C, samples were centrifuged for a period of 5 min at a centrifugation speed of 2862× p < 0.05). All experiments were performed in triplicate. One-way analysis of variance (ANOVA) was employed to assess the differences between the treatments using SPSS version 18.0 . Mean differences were determined by Tukey’s test at a confidence level of 95% (SEM images of ginseng insoluble dietary fiber (G-IDF) and enzymatically modified ginseng insoluble dietary fiber (G-MIDF) are demonstrated in −1 in both the G-IDF and G-MIDF represented –OH stretching, mainly owing to the presence of cellulose, hemicellulose, and lignin. However, the FTIR peak at the IR region of 3400 cm−1 exhibited a diminution in the G-IDF spectrum in comparison with that of a similar peak in the case of the G-MIDF at 3400 cm−1 IR region. The broadness in the IR region’s intensity for the G-MIDF could be ascribed to the breakage of intermolecular linkages. The FTIR spectra peaks showed resemblance in terms of peak shapes and IR regions, which implied that enzymatic modification did not exert any significant effect on the functional groups’ structural configurations in the case of the G-MIDF as compared to that of the G-IDF. This implied that typical functional groups of insoluble cellulose and soluble pectin exhibited stability in the G-MIDF after exposure to enzymatic hydrolysis. The strong absorption peak with a high degree of broadness at the IR region of 3419 cm−1 was mainly attributed to the presence of native cellulose and was ascribed to the functional group stretching of O–H, characterized as the molecular bonding of uronic acid. Ginseng residue has been reported to comprise rich heteropolysaccharides  and to be acidic in nature, as well as being rich in amino acids, protein, and mineral elements [−1, which was indicative of the C–H stretching of methylene (–CH2) and methyl (–CH3) functional groups, representing the typical structural configuration of cellulosic polysaccharides compounds. A minor IR peak was observed at the IR spectral region of 1741 cm−1, which was mainly due to the presence of the ester carbonyl group stretching (C=O) or acetyl (−COCH3) functional group stretching of hemicellulose. The presence of an IR peak was also recorded at the IR region of 1626 cm−1, which was ascribed to the probable presence of ionized and esterified groups of galacturonic acid. A weak peak was also observed at the IR spectral region of 1516 cm−1, along with the adjacent presence of a minor peak at 1427 cm−1; these were probably indicative of the aromatic or aliphatic C–H group IR vibrational patterns of lignin. Weak IR vibrations were recorded at 1319 cm−1 and 1242 cm−1, which were mainly attributed to the typical cellulose structure and the C–O and O–H group vibration of hemicellulose. Small peaks at IR regions of 900–1200 cm−1 as well as 1056 cm−1 were ascribed to the stretching vibration of the acyl-oxygen (CO–OR) of hemicellulose and the carbonyl (C–O) stretching vibration of the guaiacyl unit of lignin. It was evident from the above results that G-MIDF exhibited typical functional group configurations for cellulose polysaccharides, including pectin, hemicellulose, cellulose, and lignin, which was consistent with the composition of ginseng DF reported in the published literature [The FT-IR spectra for G-IDF and G-MIDF are shown in elements . Howeverterature , who eluX-ray diffractometry was employed to determine the crystal structure of the G-IDF and G-MIDF, and the results are depicted in Thermal properties in terms of weight loss for the G-IDF and G-MIDF were determined, and the results are depicted in Plant cell walls comprise lignin networks composed of ether and ester linkages with hemicellulose, whereas cellulose is usually found in an embedded form and plays a role in the formation of fiber–polymer complexes. Lignin’s amorphous structure exhibits high pyrolysis resistance to thermal processes, resulting in the consequently reduced thermal accessibility of cellulose. Generally, non-starch polysaccharides exhibit thermal stability in the following order: lignin > cellulose > hemicellulose. Regarding the pyrolysis mechanism of cellulose, the most commonly reported pathway is reported in the literature as follows: Activated cellulose is produced from cellulose with a lower degree of polymerization through complex reactions, such as dehydration, condensation, and polymerization . Then, tThe rheological behaviors of the G-MIDF and G-IDF were evaluated, and the results are tabulated in 2 m Pa·s , respectp > 0.05) effect on the apparent viscosity of the G-MIDF in comparison with the G-IDF. The consistency coefficient (K) value of the G-MIDF was significantly (p < 0.05) lower (59.51) than that of the G-IDF (67.59). This could be attributed to the destruction of ginseng DF’s molecular structure, breakage of partial branching patterns, as well as the decreasing trend in internal molecular linkages after exposure to enzymatic hydrolysis. Moreover, slight differences in the flow behavior index and apparent viscosity were observed between the G-MIDF and G-IDF. Similar findings have been reported by Jiang et al. [Panax notoginseng). Dilatant fluids are also reported in the literature as shear-thickening fluids, which usually show an increasing tendency of apparent viscosity with the corresponding rise in the shear rate . The G-Ig et al.  for dietThe hydration and functional properties of the G-IDF and G-MIDF were evaluated, and the results are tabulated in p < 0.05) WHC (6.36 g/g) in case of the G-MIDF as compared to that of the WHC of the G-IDF (5.31 g/g) (Panax notoginseng) [Sagittaria sagittifolia L.) [Enzymatic hydrolysis resulted in a significantly higher (.31 g/g) . It was ginseng) , arrowheolia L.) , and ricolia L.) . The risThe ORC values of the G-MIDF and G-IDF are given in p < 0.05) rise by 28.85% from 6.17 g/g (G-IDF) to 7.91 g/g (G-MIDF). The increasing tendency of the WSC might be ascribed to the association of the WSC with the corresponding cellulose degradation. Enzymatic hydrolysis caused the disintegration of long cellulose chains to form small cellulose polymer chains, which causes the incorporation of enlarged void spaces. Exposure to enzymatic treatment has been reported to not only cause disruption in the covalent linkages between cellulose and hemicellulose but also to lead to the breakage of ether linkages linking hemicellulose to lignin. The results of the current research were in line with findings reported by Jiang et al. [The WSC values of the G-MIDF and G-IDF are given in g et al.  for ginsp < 0.05) increase by 4.23% from 119.92 µg/g (G-IDF) to 124.28 µg/g (G-MIDF). Likewise, at pH 7.0, the NAC increased from 117.17 µg/g (G-IDF) to 122.29 µg/g (G-MIDF). Hence, enzymatic hydrolysis improved the NAC at both pH levels. Similarly, the increasing tendencies in NAC have also been reported by Luo et al. [2–)has been reported in the processing of cured meat products for rendering development of unique colors and characteristic flavors, as well as pathogen inhibition [N-nitroso compounds after coming into contact with amides and secondary amines (under acidic conditions), which exhibit carcinogenicity. The NAC is regarded as one of DF’s most essential properties, which may play a contributing role in lowering toxicity to humans. Therefore, in order to overcome this adverse effect pertaining to cured meat products, phyto-chemicals with high NACs could be employed, and G-MIDF with improved NAC might be incorporated into reformulated meat products [The NAC values of the G-MIDF and G-IDF are given in o et al.  in the co et al. . Moreovehibition . Similarproducts . The BAC values of the G-MIDF and G-IDF are given in p < 0.05) rise by 24.15% from 8.86 mg/g (G-IDF) to 10.85 mg/g (G-MIDF). After exposure to the enzymatic hydrolysis, the G-MIDF exhibited significant improvement in the CAC. The possible reason for the enhanced CAC in case of the G-MIDF might be ascribed to the loosening of the IDF’s structural framework due to enzymatic hydrolysis, which led to an increased degree of exposure of hydrophobic groups in comparison with the G-IDF. Furthermore, it may be implied that the higher CAC values of the G-MIDF were because of rising tendencies in various properties of the G-MIDF, including specific surface area and viscosity, after exposure to enzymatic hydrolysis, which led to the formation of more complex structures and resulted in the increased binding of cholesterol [The CAC is regarded as an important parameter for the evaluation of IDF’s ability to absorb lipophilic components both under conditions of the intestinal environment and in stimulated stomachs. The CAC values of the G-MIDF and G-IDF are provided in lesterol ,35. Theslesterol , peach flesterol , and whelesterol . p < 0.05) absorption ability in terms of binding cholesterol after exposure to enzymatic hydrolysis at all concentrations. The probable mechanism for enhanced GAC might be ascribed to glucose absorption by IDF, which may possibly occur because of the physical barrier as well as the chemisorption and ensnarement of glucose molecules. Most probably, the enhanced availability of cavities and surface areas due to the structural changes caused by enzymatic hydrolysis may help in achieving a greater entrapment of glucose molecules [The GAC values of the G-MIDF and G-IDF are given in olecules . Furtherolecules  in the cThis study investigated the effects of enzymatic modification using cellulase/xylanase on the composition and structural and functional properties of ginseng insoluble dietary fiber (IDF). Fourier transform infrared spectroscopy, X-ray diffractometry, and scanning electron microcopy showed significant structural alterations in the ginseng-modified (G-MIDF) samples, which exhibited more porous and completely wrinkled surfaces. Thermo-gravimetric analysis showed that the G-MIDF exhibited a specified range of pyrolysis temperatures, making it suitable for food applications involving processing at temperatures below 300 °C. The G-IDF had relatively lower apparent viscosity in comparison with the enzymatically modified DF (G-MIDF). Overall, it was evident from the rheograms that both the G-IDF and G-MIDF exhibited a resemblance with respect to viscosity changes as a function of shear rate. Enzymatic treatment led to significant improvement in hydration and functional properties, such as water holding, oil retention, water swelling, nitrite ion binding, bile acid binding, cholesterol absorption, and glucose absorption capacities.


Glycyrrhiza species, possesses a wide range of therapeutic applications, including antiviral properties. Glycyrrhizic acid (GL) and glycyrrhetinic acid (GA) are the most important active ingredients in licorice. Glycyrrhetinic acid 3-O-mono-β-d-glucuronide (GAMG) is the active metabolite of GL. GL and its metabolites have a wide range of antiviral activities against viruses, such as, the hepatitis virus, herpes virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and so on. Although their antiviral activity has been widely reported, the specific mechanism of action involving multiple links such as the virus itself, cells, and immunity are not clearly established. In this review, we will give an update on the role of GL and its metabolites as antiviral agents, and detail relevant evidence on the potential use and mechanisms of actions. Analyzing antivirals, their signaling, and the impacts of tissue and autoimmune protection may provide promising new therapeutic strategies.Licorice, a natural medicine derived from the roots and rhizomes of  O-mono-β-d-glucuronide (GAMG). GAMG, a distal glucuronic acid hydrolysate of GL, has a higher bioavailability and stronger physiological functions than GL, including antitumor, antiviral and anti-inflammatory activities [Glycyrrhizic acid (GL), a tetracyclic triterpenoid saponin, has a broad spectrum of biological activities , especiativities ,7. In paViruses usually include plant viruses, animal viruses and bacterial viruses. It is an organism with a special structure, usually containing only DNA or RNA, and parasitic in living cells, multiplying by self-replication. However, many viruses have a strong ability to invade, destroy, and even endanger the health of humans, animals, and plants. For example, the new SARS-CoV-2 virus that broke out in 2019 has been an ever-present threat to public health worldwide, and has already resulted in millions of dead. The constant mutation of the virus has brought great difficulties to epidemic prevention and people’s treatment. In addition, there are many common viruses in daily life, such as hepatitis viruses, influenza viruses, and herpesviruses.Viral proliferation is a complex process, and the antiviral effect of natural products may influence the entry , gene replication, and exit  . The antIn this review, two investigators conducted an independent literature search using the PubMed, Embase, Cochrane Library, and Web of Science databases from the databases’ inception to 31 July 2022 (Jiawei Zuo and Tao Meng). The following key terms were used: “glycyrrhizic acid”, “glycyrrhetinic acid”, “Glycyrrhizin monosaccharides”, and “antiviral”. The cited references for the relevant systematic reviews and conference proceedings were manually cross-checked to identify additional literature. Articles pertinent to antiviral activity of glycyrrhizic acid and its metabolites were selected for this review. Searches were not restricted by language, country, or publication date. Articles were initially screened based on the title and abstract reading, and then full texts of potentially relevant publications were obtained and reviewed by two authors independently (Jiawei Zuo and Tao Meng) to determine the publications’ eligibility. Any discrepancies were resolved by discussion, and a third assessor arbitrated any disagreements.The purpose of this article is to comprehensively review the safety and efficacy of GL and its metabolites for antiviral effects. Complex mechanisms of action are elaborated to highlight major therapeutic targets that may help reduce the damage caused by various viruses. This article provides a narrative and critical review of the clinical efficacy and pharmacological effects of GL and its metabolites in various viral infections, keeping clinicians and researchers informed of the latest results and main roles. In addition, it provides an overview of some complexes containing GL and its metabolites, and the application of novel delivery regimes that contribute to a better understanding of their use and optimization.In 2019, the Coronaviruse disease (COVID-19) broke out, caused large-scale human infection worldwide, and brought serious health hazards and economic losses. So far, the virus has continued to mutate and spread, and it is still seriously endangering human health and safety. At present, the SARS-CoV-2 infection cannot be effectively inhibited after vaccination, and there is no effective drug for the treatment of the viral infection in clinical practice. It was previously reported that ACE2 was proven to be an important and special receptor for the SARS coronavirus . Recent SARS-CoV-2 is a β-coronavirus which contains three transmembrane proteins, spike protein (S), membrane protein (M), and envelope protein (E). SARS-CoV-2 also contains a large nucleoprotein (N) enveloped positive RNA genome . GA has Virtual screening also revealed that GL is effective against the target proteins of SARS-CoV-2 , serves The various pharmacological activities of GL and its derivatives were summarized, and the structurally modified GL-related derivatives were further developed to make them less cytotoxic and more targeted . MoreoveThe RBD in the SARS-CoV-2 nucleocapsid protein may be a potential drug target that plays a variety of key functions in the life cycle of the virus, especially viral replication. GL showed a higher binding affinity with target proteins, which may help block the important site within the RNA binding domain of SARS-CoV-2 viral nucleocapsid protein (N) and reduce the risk of infection in the host . Viral RIn addition, virtual screening showed that GL had the best affinity towards key proteases of SARS-CoV-2, which played a pivotal role in mediating viral replication and transcription ,39. It hFrom a pharmacological point of view, GL showed therapeutic potential for COVID-19 through binding to ACE2, down-regulating proinflammatory cytokines, inhibiting intracellular reactive oxygen species (ROS) accumulation, suppressing high respiratory tract output, and inducing endogenous interferons . The antIn more recent years, there have been an increasing number of studies on the activity of GAMG. During the process of the LPS-induced RAW264.7 cell inflammatory response, GAMG showed a higher anti-inflammatory activity than GL, which may be due to the stronger inhibitory effect of GAMG on IL-6, iNOS, and COX-2. In-depth research on the anti-inflammatory mechanism of GAMG found that it could block nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPKs) signaling pathways ,47. HighThe long-term medical GL and its preparations made GL a good candidate against SARS-CoV-2 . The comThe antiviral activity of GA has also been extensively studied. Due to high safety as a natural sweetener, GA is widely used as a clinical therapeutic . At presThe liver is an organ with mainly metabolic functions in the body and plays a role in deoxidation, storage of glycogen, and synthesis of secreted proteins. Liver disease accounts for approximately 2 million deaths per year worldwide, due to complications from cirrhosis, and viral hepatitis and hepatocellular carcinoma (HCC). GL and its derivatives may have a therapeutic value in the hepatitis disease caused by viral infections. Some glycyrrhizin treatment mechanisms and references for various hepatitis viruses are shown in The hepatitis B virus (HBV) infection is one of the major public health problems worldwide, about 30% of the world’s population shows serological evidence of past or present infection . It is gIn 2017, researchers were surprised to find that GA had a synergistic effect with entecavir for the therapy of HBV infection . GA inflThe hepatitis C virus (HCV) infection is an important cause of chronic hepatitis C, cirrhosis and HCC worldwide . GL inhiFurthermore, GL intravenous injection obviously decreased the incidence of HCC in HCV-infected patients . GL alonThe hepatitis E virus (HEV) is an enterically transmitted virus . GloballHerpesviruses are large, diverse and have a double-stranded DNA (dsDNA) molecule wrapped in a rigid protein shell. They contain a wide variety, such as herpes simplex virus 1 (HSV-1) and HSV-2, EBV, varicella zoster virus (VZV), human cytomegalovirus (HCMV), human herpesvirus-6 (HHV-6), Human herpesvirus-7 (HHV-7), and Kaposi’s sarcoma-associated herpesvirus (KSHV) . In geneHSV-1 can cause severe diseases among people with low immunity. Under normal circumstances, T cell activity that is related to cell-mediated immunity plays an important defense against herpesvirus infections. Researches indicated that the mortality rate of special pathology mice exposed to HSV decreased from 80% to 5% after GL treatment, which significantly improved survival in HCV-infected mice . These rGL and its derivatives were capable of hindering the growth and cytopathic effects of HSV ,91. In aVZV is a globally infectious human virus that causes chickenpox and is usually spread through broken skin or respiratory droplets . Under cEBV is a well-known pathogen of infectious mononucleosis, and many diseases are often related to EBV in etiology. Many types of cancer are accompanied by dysregulated sumoylation processes ,103,104.KSHV is the cause of Kaposi’s sarcoma, multicentric Castleman disease, and primary exudative lymphoma. Patients treated with GL had lower KSHV DNA levels than those without therapy . GL, whiThe activation of any TLR would result in the production of interferon and the upregulation of cytokines and chemokines . HoweverGL might affect the maturation of mRNA encoding LANA and selectively inhibit the growth of KSHV-infected lymphocytes . The stuClinical practice has shown that GL treatment of patients with hemophilia or with acquired immunodeficiency syndrome could increase the level of the p24 antigen of the human immunodeficiency virus-1 (HIV-1), suggesting that GL may inhibit HIV-1 replication in vivo . GL not There are some clinical data showing that in cultures of 12 patient peripheral blood mononuclear (PBM) cell samples infected with a slow-replicating nonsyncytium-inducing variant of HIV (NSI-HIV), GL inhibited HIV replication by more than 90% . DiffereAfter the virus particles attach and adsorb to the specific receptor on the host cell, the enveloped virus must pass its own lipid bilayer envelope coating and the plasma cell membrane lipid bilayer fusion layer through the fusion pore to enter the host. The fluidity of the HIV-1 envelope was always lower than that of the plasma membrane because the cholesterol/protein ratio of the viral envelope was higher than that of the plasma membrane ,123. DecThe influenza virus and respiratory syncytial virus infection have become the main killers for viral respiratory diseases, and Chinese herbal medicines can inhibit virus reproduction, enhance human immunity and so on, which have good antiviral activity. The earlier studies showed that the treatment of mice infected with 10–50% of the lethal dose of the influenza A2 virus with GL exhibited promising therapeutic efficacy . GL can GL had little effect on H5N1 replication and apoptosis of infected cells to a certain extent but diminished the migration of monocytes to the supernatant of A549 cells infected with H5N1 . GL, at The triterpenoid saponin isolated from licorice contained a saccharide moiety and glucuronic acid or galacturonic acid and showed good inhibitory activity against the H1N1 influenza virus in MDCK cells . MoreoveMost reports indicate that antiviral activity is one of the broad pharmacological activities of GL. GL is only active at high non-cytotoxic concentrations, but different concentrations of GL can effectively inhibit the replication of flaviviruses . LikewisGL neither directly inactivated porcine reproductive and respiratory syndrome virus (PRRSV) particles nor inhibited the release of PRRSV. Among them, both PRRSV proliferation and the expression of PRRSV-encoded proteins were reduced. In terms of the mechanism, GL mainly inhibited the infiltration phase and had little influence on the PRRSV adsorption or release steps . GL alsoGL can effectively inhibit the cytopathic effect (CPE) induced by dengue virus 2 and reduce the infectivity of the virus at the same time . The intCarbon dots (CDs) are increasingly used in the field of antiviral research. This was a new attempt to combine CDs with GL to develop not only high biocompatibility but also excellent antiviral activity. Using PRRSV as a model, GL-based carbon dots (GL-CDs) activity detection was carried out . DetaileGL can treat the SARS-CoV-2 infection through interacting with ACE2 to prevent the viral replication and spread, interacting with the spike protein to block the viral binding and fusion, inhibiting the envelope protein to disrupt the viral function and structure, and targeting the RNA binding domain of nucleocapsid protein to prevent the viral replication. GL can also exert the therapeutic potential for COVID-19 through down-regulating pro-inflammatory cytokines, inhibiting ROS accumulation, decreasing thrombin, suppressing high respiratory tract output and inducing endogenous interferon. Physiology-based pharmacokinetic studies indicated that GAMG can be absorbed quickly and effectively, and exert better pharmacological activities than GL. GAMG exhibited benign drug-like properties due to higher blood concentration and higher tissue distribution in the lungs and kidney . GAMG caFurther, molecular docking showed that GAMG binds to the non-structural protein 3CLpro of SARS-CoV-2, but GL and GA do not, indicating that GAMG may have a special and good activity for inhibiting SARS-CoV-2 A. MoreovThe understanding of the interactions between GAMG, GL, GA and infectious factors can improve the remedy of COVID-19 patients and reduce the risk of antiviral resistance . All in GL and GL-containing drugs have been widely used in clinical practice. GA and GAMG are the active metabolites of GL. GL and GA exhibit antiviral activity by affecting the proliferation and pathogenic effects of various viruses. Here, we summarized the action mechanism of GL, GA and GAMG against various viruses, including SARS-CoVs, hepatitis virus, and herpesvirus. GL and GA not only apply direct chemotherapy to inhibit various pathogenic viruses but also can interact with cell membranes or exert joint immunity with the host. GL can suppress gene expression of potential oncogenic viruses to treat severe and incurable chronic viral diseases, such as Kaposi’s sarcoma. GL can treat SARS-CoV-2 during and after virus adsorption through inhibiting virus adsorption and penetration. GL exerted the therapeutic potential for SARS-CoV-2 by combination with ACE2, down-regulating pro-inflammatory cytokines, decreasing the accumulation of ROS, inhibiting thrombin and airway exudates, and inducing endogenous interferon. As promising antiviral drug candidates, GL, GA and GAMG can inhibit virus replication and eliminate latent virus to combat the increasing variety and destructive power of viruses. Interestingly, pharmacokinetics and molecular docking showed that GAMG with benign drug-like properties can more efficiently target ACE2 and remedy SARS-CoV-2 infection than GL. Thus, GAMG and GL are expected to become broad-spectrum antiviral agents and be widely used in clinical treatments.