Wolffia Globosa Research Articles

Nutritional Quality, Functional Properties, and Biological Characterization of Watermeal (Wolffia globosa)

Watermeal (Wolffia globosa) has emerged as a promising candidate as an alternative protein source due to its outstanding nutritional profile. This study evaluated the nutritional quality, functional properties, and biological characteristics of W. globosa. The protein content was found to be high at 26.76 g/100 g of dry weight (DW). Functional properties, including water and oil holding capacities, protein solubility, foaming, and emulsifying abilities across different pH levels, were assessed to determine optimal conditions. Amino acid profiling revealed that essential amino acids constitute 67% of the total amino acids. Polyunsaturated fatty acids (PUFAs), particularly α-linolenic (18:3 n-3) and linoleic acids (18:2 n-6), were identified as the predominant fatty acids. Additionally, watermeal was found to contain significant amounts of α-tocopherol (699 µg/g) and γ-oryzanol (100 µg/g), which contribute to its antioxidant capacity, as confirmed by DPPH and FRAP assays. The prominent protein bands were observed at around 62–67 kDa, likely representing a structural, metabolic protein (vicilin-like protein) of 38% of the total protein content. Overall, W. globosa demonstrates significant potential as a functional food ingredient, offering a rich nutrient profile and health-promoting properties, making it a promising choice for future diets.

Characterization of Bioactive Metabolites and Antioxidant Activities in Solid and Liquid Fractions of Fresh Duckweed (Wolffia globosa) Subjected to Different Cell Wall Rupture Methods.

Fresh Wolffia globosa, the smallest flowering plant well-known for its favorable nutrient composition and rich content of bioactive compounds, was subjected to boiling, freeze-thawing, and mechanical crushing to reduce its excessive (95-96%) moisture level and consequent drying time. The resultant three wolffia matrixes were filtered through a plankton net to fractionate into the residue and the filtrate. The proximate composition, bioactive metabolites, antioxidant activity, and characterization of bioactive metabolites by LC-ESI-QTOF-MS/MS and Fourier transform infrared spectroscopy were made from oven-dried residues and filtrates. Among residues, crude protein (29.84%), crude lipid (5.77%), total carotenoids (TCC; 722.8 μg/g), and vitamin C (70.02 mg/100 g) were the highest (p < 0.05) for freeze-thawing against higher ash (7.99%), total phenolic content (TPC; 191.47 mg GAE g-1 dry weight), total flavonoid content (TFC; 91.54 mg QE g-1 dry weight), DPPH activity (47.46%), and ferric reducing antioxidant power (FRAP) activity (570.19 μmol FeSO4 equiv/mg) for the crushed counterpart and Chl-b in residues from boiling. No significant variation was evident in the total tannin content (TTC). Among filtrates, higher total phenolic content (773.29 mg GAE g-1 dry weight), TFC (392.77 mg QE g-1 dry weight), TTC (22.51 mg TAE g-1), and antioxidant activity as DPPH activity (66.46%) and FRAP (891.62 μmol FeSO4 equiv/mg) were evident for boiling, while that from crushing exhibited the highest TCC (1997.38 μg/g DM). LC-ESI-QTOF-MS/MS analysis identified 72 phenolic compounds with the maximum in residue (33) and filtrate (33) from freeze-thawing, followed by crushing (18 and 19) and boiling (14 and 13) in order, respectively. The results indicated that the predrying cell rupturing method significantly impacted quantitative, as well as qualitative compositions of residues and filtrates from fresh wolffia.

Characterization of the complete chloroplast genome of Wolffia arrhiza and comparative genomic analysis with relative Wolffia species

Lemnoideae, commonly referred to as the duckweed, are aquatic plants found worldwide. Wolffia species are known for their extreme reduction in size and complexity, lacking both roots and leaves, and they hold the distinction of being the smallest plants among angiosperms. Interestingly, it belongs to the Araceae family, despite its apparent morphological differences from land plants in the same family. Traditional morphological methods have limitations in classifying these plants, making molecular-level information essential. The chloroplast genome of Wolffia arrhiza is revealed that a total length of 169,602 bp and a total GC content of 35.78%. It follows the typical quadripartite structure, which includes a large single copy (LSC, 92,172 bp) region, a small single copy (SSC, 13,686 bp) region, and a pair of inverted repeat (IR, 31,872 bp each) regions. There are 131 genes characterized, comprising 86 Protein-Coding Genes, 37 Transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. Moreover, 48 simple sequence repeats and 32 long repeat sequences were detected. Comparative analysis between W. arrhiza and six other Lemnoideae species identified 12 hotspots of high nucleotide diversity. In addition, a phylogenetic analysis was performed using 14 species belonging to the Araceae family and one external species as an outgroup. This analysis unveiled W. arrhiza and Wolffia globosa as closely related sister species. Therefore, this research has revealed the complete chloroplast genome data of W. arrhiza, offering a more detailed understanding of its evolutionary position and phylogenetic categorization within the Lemnoideae subfamily.

Synergistic activity of Limosilactobacillus reuteri KUB-AC5 and water-based plants against Salmonella challenge in a human in vitro gut model.

A synbiotic is a combination of live microorganisms and specific substrates that are selectively utilized by host microorganisms, resulting in health benefits for the host. Previous studies have demonstrated the protective effects of L. reuteri KUB-AC5 against Salmonella infection in chicken and mouse models. The probiotic activity of L. reuteri KUB-AC5 in these hosts was influenced by nutritional supplements. Water-based plants contain significant amounts of carbohydrates, particularly dietary fiber and proteins, making them potential prebiotic substrates. In this study, four water-based plants (Ulva rigida, Caulerpa lentillifera, Wolffia globosa, and Gracillaria fisheri) were screened for their ability to support the growth of L. reuteri KUB-AC5. Under monoculture testing, U. rigida exhibited the highest capacity to support the growth of L. reuteri KUB-AC5 and the production of organic acids, including acetic acid, lactic acid, and propionic acid (p ≤ 0.05). In co-culture experiments, the synbiotic combination of U. rigida and L. reuteri KUB-AC5 demonstrated the potential to eliminate Salmonella Typhimurium DMST 48437 when inoculated at 104CFU/mL within 9h. The synbiotic activities of U. rigida and L. reuteri KUB-AC5 were further investigated using an in vitro human gut model. Compared to the probiotic treatment, the synbiotic combination of L. reuteri KUB-AC5 and U. rigida showed significantly higher levels of L. reuteri KUB-AC5 (5.1 log copies/mL) and a reduction of S. Typhimurium by 0.8 log (CFU/ml) after 24h (p ≤ 0.05). Synbiotic treatment also significantly promoted the production of short-chain fatty acids (SCFAs), including butyric acid, propionic acid, and acetic acid, compared to prebiotic and probiotic treatments alone (p ≤ 0.05). Furthermore, the synbiotic formulation modulated the in vitro simulated gut microbiome, enhancing putatively beneficial gut microbes, including lactobacilli, Faecalibacterium, and Blautia. Our findings demonstrated that L. reuteri KUB-AC5, in combination with U. rigida, exhibited synergistic activity, as indicated by increased viability, higher anti-pathogenicity toward Salmonella, and the ability to modulate the gut microbiome.

Wolffia globosa (Araceae s. l. / Lemnaceae s. str.), a new aquatic alien species in the flora of Ukraine

In Europe, Wolffia globosa was first recorded in 2002 in the RF, and not in Bulgaria, as it was previously believed. Over the past 20 years, the species has expanded its secondary range. As of 2023, it is known from seven European countries. In Ukraine, W. globosa was first found in 2023 in two localities in Zhytomyr Region (Zhytomyr Polissia). The plant communities with W. globosa in Ukraine belong to the association Lemno-Spirodeletum polyrhizae of the alliance Lemnion minoris of the class Lemnetea. In the reported localities W. globosa occurs in eutrophic stagnant waterbodies with depths of 0.2–0.5 m and weakly flowing waterbodies with depths of 1.2–1.5 m; silty bottom is covered by a silt layer 5–20 cm thick, lighting is full. It is suggested that due to the observed climate warming, further migrations of W. globosa to other regions of Ukraine are possible.

Effects of altered gravity on growth and morphology in Wolffia globosa implications for bioregenerative life support systems and space-based agriculture

Understanding the response of plants to varied gravitational conditions is vital for developing effective food production in space bioregenerative life support systems. This study examines the impact of altered gravity conditions on the growth and morphological responses of Wolffia globosa (commonly known as “water lentils” or “duckweed”), assessing its potential as a space crop. Although an experiment testing the effect of simulated microgravity on Wolffia globosa has been previously conducted, for the first time, we investigated the effect of multiple gravity levels on the growth and morphological traits of Wolffia globosa plants. The plant responses to simulated microgravity, simulated partial gravity (Moon), and hypergravity environments were evaluated using random positioning machines and the large-diameter centrifuge. As hypothesized, we observed a slight reaction to different gravitational levels in the growth and morphological traits of Wolffia globosa. The relative growth rates (RGR) of plants subjected to simulated microgravity and partial gravity were reduced when compared to those in other gravity levels. The morphological analysis revealed differences in plant dimensions and frond length-to-width ratios under diverse gravity conditions. Our findings showed that Wolffia globosa is responsive to gravitational changes, with its growth and morphological adaptations being slightly influenced by varying gravitational environments. As for other crop species, growth was reduced by the microgravity conditions; however, RGR remained substantial at 0.33 a day. In conclusion, this study underscores the potential of Wolffia globosa as a space crop and its adaptability to diverse gravitational conditions, contributing to the development of sustainable food production and bioregenerative life support systems for future space exploration missions.

The effect of polyphenols on DNA methylation-assessed biological age attenuation: the DIRECT PLUS randomized controlled trial

BackgroundEpigenetic age is an estimator of biological age based on DNA methylation; its discrepancy from chronologic age warrants further investigation. We recently reported that greater polyphenol intake benefitted ectopic fats, brain function, and gut microbiota profile, corresponding with elevated urine polyphenols. The effect of polyphenol-rich dietary interventions on biological aging is yet to be determined.MethodsWe calculated different biological aging epigenetic clocks of different generations (Horvath2013, Hannum2013, Li2018, Horvath skin and blood2018, PhenoAge2018, PCGrimAge2022), their corresponding age and intrinsic age accelerations, and DunedinPACE, all based on DNA methylation (Illumina EPIC array; pre-specified secondary outcome) for 256 participants with abdominal obesity or dyslipidemia, before and after the 18-month DIRECT PLUS randomized controlled trial. Three interventions were assigned: healthy dietary guidelines, a Mediterranean (MED) diet, and a polyphenol-rich, low-red/processed meat Green-MED diet. Both MED groups consumed 28 g walnuts/day (+ 440 mg/day polyphenols). The Green-MED group consumed green tea (3–4 cups/day) and Mankai (Wolffia globosa strain) 500-ml green shake (+ 800 mg/day polyphenols). Adherence to the Green-MED diet was assessed by questionnaire and urine polyphenols metabolomics (high-performance liquid chromatography quadrupole time of flight).ResultsBaseline chronological age (51.3 ± 10.6 years) was significantly correlated with all methylation age (mAge) clocks with correlations ranging from 0.83 to 0.95; p < 2.2e − 16 for all. While all interventions did not differ in terms of changes between mAge clocks, greater Green-Med diet adherence was associated with a lower 18-month relative change (i.e., greater mAge attenuation) in Li and Hannum mAge (beta = − 0.41, p = 0.004 and beta = − 0.38, p = 0.03, respectively; multivariate models). Greater Li mAge attenuation (multivariate models adjusted for age, sex, baseline mAge, and weight loss) was mostly affected by higher intake of Mankai (beta = − 1.8; p = 0.061) and green tea (beta = − 1.57; p = 0.0016) and corresponded with elevated urine polyphenols: hydroxytyrosol, tyrosol, and urolithin C (p < 0.05 for all) and urolithin A (p = 0.08), highly common in green plants. Overall, participants undergoing either MED-style diet had ~ 8.9 months favorable difference between the observed and expected Li mAge at the end of the intervention (p = 0.02).ConclusionsThis study showed that MED and green-MED diets with increased polyphenols intake, such as green tea and Mankai, are inversely associated with biological aging. To the best of our knowledge, this is the first clinical trial to indicate a potential link between polyphenol intake, urine polyphenols, and biological aging.Trial registrationClinicalTrials.gov, NCT03020186.

Wolffia globosa-Based Nutritious Snack Formulation with High Protein and Dietary Fiber Contents.

Wolffia globosa (W. globosa) or duckweed is a small aquatic plant with high protein, dietary fiber, and lipid contents that can be combined with food products to develop nutritious snacks as one strategy to mitigate malnutrition. Here, response surface methodology (RSM) with mixture design was used to develop snacks from W. globosa freeze-dried powder (WP). The physical properties, proximate analysis, amino acid profiles, sensory evaluation, phytochemical analysis, antioxidant properties, and genotoxicity (Ames test) of the snacks were evaluated. The optimal W. globosa snack formula was 64% glutinous rice flour, 10% tapioca flour, and 26% WP, giving a highly desirable liking score of 1.00. Addition of WP increased crude protein, essential amino acids, and dietary fiber compared with the control snack by 51%, 147%, and 83%, respectively. According to the Thai recommended daily intakes, the developed W. globosa snack had high protein and dietary fiber. Phytochemical contents and antioxidant activities of the W. globosa snack such as total phenolic contents (TPCs), total flavonoid contents (TFCs), ferric ion reducing antioxidant power (FRAP) activity, and oxygen radical absorbance capacity (ORAC) activity were significantly higher than the control snack. The novel combination of WP with snack product ingredients greatly enhanced nutritional value.

Effect of fertilization rates on metabolites of duckweed (Wolffia globosa) as potential pharmaceutical resource

Effect of fertilization rates on metabolites of duckweed (Wolffia globosa) as potential pharmaceutical resource

The effect of high-polyphenol Mediterranean diet on visceral adiposity: the DIRECT PLUS randomized controlled trial

BackgroundMediterranean (MED) diet is a rich source of polyphenols, which benefit adiposity by several mechanisms. We explored the effect of the green-MED diet, twice fortified in dietary polyphenols and lower in red/processed meat, on visceral adipose tissue (VAT).MethodsIn the 18-month Dietary Intervention Randomized Controlled Trial PoLyphenols UnproceSsed (DIRECT-PLUS) weight-loss trial, 294 participants were randomized to (A) healthy dietary guidelines (HDG), (B) MED, or (C) green-MED diets, all combined with physical activity. Both isocaloric MED groups consumed 28 g/day of walnuts (+ 440 mg/day polyphenols). The green-MED group further consumed green tea (3–4 cups/day) and Wolffia globosa (duckweed strain) plant green shake (100 g frozen cubes/day) (+ 800mg/day polyphenols) and reduced red meat intake. We used magnetic resonance imaging (MRI) to quantify the abdominal adipose tissues.ResultsParticipants (age = 51 years; 88% men; body mass index = 31.2 kg/m2; 29% VAT) had an 89.8% retention rate and 79.3% completed eligible MRIs. While both MED diets reached similar moderate weight (MED: − 2.7%, green-MED: − 3.9%) and waist circumference (MED: − 4.7%, green-MED: − 5.7%) loss, the green-MED dieters doubled the VAT loss (HDG: − 4.2%, MED: − 6.0%, green-MED: − 14.1%; p < 0.05, independent of age, sex, waist circumference, or weight loss). Higher dietary consumption of green tea, walnuts, and Wolffia globosa; lower red meat intake; higher total plasma polyphenols (mainly hippuric acid), and elevated urine urolithin A polyphenol were significantly related to greater VAT loss (p < 0.05, multivariate models).ConclusionsA green-MED diet, enriched with plant-based polyphenols and lower in red/processed meat, may be a potent intervention to promote visceral adiposity regression.Trial registrationClinicalTrials.gov, NCT03020186

Total Active Compounds and Mineral Contents in Wolffia globosa

Wolffia globosa, or watermeal, is an aquatic plant belonging to the Lemnaceae family that is consumed as food and sold in local markets of Thailand. The aim of this study was to quantify selected active compounds and minerals in W. globosa ethanolic extract and evaluate its antioxidant activity. Total phenolic, flavonoid, and anthocyanin contents were analyzed. High-performance liquid chromatography was used for the determination of beta-carotene, ferulic acid, luteolin-7-O-β-D-glucoside, and kaempferol. Mineral contents (iron, potassium, calcium, magnesium, zinc, and sodium) were determined by atomic absorption spectroscopy. Antioxidative activity was evaluated by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azobis (3-ethylbenzothiazoline-6-sulfonic acid)) radical scavenging assays. The beta-carotene, ferulic acid, luteolin-7-O-β-D-glucoside, and kaempferol contents of the extract were 2.52 ± 0.10, 1.40 ± 0.10, 2.42 ± 0.50, and 1.57 ± 0.14 mg/g extract, respectively. The highest mineral content in the W. globosa extract was magnesium. The wet extract of W. globosa showed higher amounts of all minerals than the dry extract. Freshly prepared and boiled W. globosa extracts showed radical scavenging activity at 1000 µg/milliliter with 75.77 ± 0.93% and 67.10 ± 0.20% inhibition of DPPH and 70.40 ± 7.20% and 59.78 ± 3.16% inhibition of ABTS, respectively. This plant is a promising novel source of natural phytochemical constituents and antioxidants and has potential for development as a plant-based nutraceutical product for the treatment of diseases caused by free radicals.

Antimicrobial and Functional Properties of Duckweed (Wolffia globosa) Protein and Peptide Extracts Prepared by Ultrasound-Assisted Extraction.

Wolffia globosa is an interesting alternative plant-based protein source containing up to 40% protein dry weight. Dried duckweed protein extract (PE) was obtained using ultrasound-assisted extraction (UAE) before isoelectric precipitation (pH 3.5) to yield protein concentrate (PC) and protein solution (PS). The PC was hydrolyzed using Alcalase enzyme to obtain protein concentrate hydrolysate (PCH). Among all fractions, PCH exhibited antimicrobial properties by decreasing populations of Vibrio parahaemolyticus and Candida albicans at 0.43 ± 1.31 log reduction (66.21%) and 3.70 ± 0.11 log reduction (99.98%), respectively. The PE and PS also showed high solubilities at pH 8 of 90.49% and 86.84%, respectively. The PE demonstrated the highest emulsifying capacity (EC) (71.29%) at pH 4, while the highest emulsifying stability (ES) (~98%) was obtained from the PE and PS at pH 6 and pH 2, respectively. The major molecular weights (Mw) of the PE, PC, PCH and PS were observed at 25, 45, 63 and 100 kDa, with a decrease in the Mw of the PCH (<5 kDa). The PCH contained the highest total amino acids, with aspartic acid and glutamic acid being the major components. The results revealed the antimicrobial and functional properties of duckweed protein and hydrolysate for the first time and showed their potential for further development as functional food ingredients.

Freshwater Macrophytes: A Potential Source of Minerals and Fatty Acids for Fish, Poultry, and Livestock.

The freshwater macrophytes are abundant in tropical and subtropical climates. These macrophytes may be used as feed ingredients for fish and other animals. The nutritional value of twelve freshwater-cultured macrophytes was evaluated in the present study. Significantly higher crude protein (36.94–36.65%) and lipid (8.13–7.62%) were found in Lemna minor and Spirodela polyrhiza; ash content was significantly higher in Hydrilla verticillata, Wolffia globosa, and Pistia stratiotes (20.69–21.00%) compared with others. The highest levels of sodium, magnesium, chromium, and iron levels were recorded in P. stratiotes. H. verticillata was a rich source of copper, manganese, cobalt, and zinc; the contents of calcium, magnesium, strontium, and nickel were highest in S. polyrhiza. Selenium and potassium contents were higher in Salvinia natans and W. globosa, respectively. The n-6 and n-3 polyunsaturated fatty acids (PUFAs) contents were significantly higher in W. globosa and Ipomoea aquatica, respectively compared with others. Linoleic and α-linolenic acids were dominant n-6 and n-3 PUFAs. The highest value (4.04) of n-3/n-6 was found in I. aquatica. The ratio ranged from 0.61 to 2.46 in other macrophytes. This study reveals that macrophytes are rich sources of minerals, n-6 and n-3 PUFAs.

Duckweed as a future food: Evidence from metabolite profile, nutritional and microbial analyses

Duckweed species are nutritionally meaningless plant which grow wildly in unattended areas. Therefore, understanding the metabolites content in duckweed species is essential for designing a future food products. Here, we report an untargeted Gas Chromatography-Mass Spectrometry (GC-MS)-based metabolomics approach for comprehensively discriminating between Lemna minor and Wolffia globosa of duckweeds species using principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). Ten differential metabolites levels were tentatively identified between L. minor and W. globosa. Relative to W. globosa, L. minor appeared to enrich with 5-Hydroxyl-L-tryptophan, Tocopheryl acetate, Naringenin, α-linolenic acid and glutamic acid. Furthermore, the nutritional and microbial analyses of ice cream formulated with dried L. minor were investigated. The nutritional analysis results show that relative to control, the ice cream with 2% dried L. minor had significantly increased protein, fiber and ash content. In addition, total plate count (TPC) for microbial analysis of duckweed ice cream was performed. The result suggested that the small amount of bacteria (3.82 cfu/g) was traced in formulated ice cream with 2% dried L. minor. Overall, the metabolites profile, nutritional and microbial analyses of food used L. minor plant indicate that duckweed is a good candidate for future food.

Nutrition Value and Growth Ability of Aquatic Weed Wolffia globosa as Alternative Feed Sources for Aquaculture System

One of the problems of aquaculture is the high cost of feed. Therefore, how to minimize the feed cost is of the main goals. It is necessary to find out natural sources that can be used as alternative feed. Wolffia globosa is an aquatic weed that is presumed to be used as an alternative feed source. Study the nutritional content of the Wolffia was conducted in June 2019, while the growth ability was conducted both outdoors and indoors in July 2019 at Research Center for Limnology Laboratory, LIPI. The W. globosa (percentage in dry weight) contained protein: 45.54%, fat: 5.33%, crude fiber: 9.98%, ash: 20.43%, and Nitrogen Free Extract:19.21%. It also contains 15 types of amino acids as much as 1.51 %w/w. At the outdoor system culture, the Wolffia has daily growth rate/GR (g/day) was 64.62−120.6, specific growth rate/SGR (%/day) was 14.622-15.333, and the daily productivity/P (g/m2/day) was 97.757−176.659. While at the indoor system culture the value of GR (g/day), SGR (%/day), and P (g/m2/day) were 0.675−1.50; 9.889−12.643; and 25.883−64.564 respectively. It is indicated that this Wolffia is very potential to be an alternative natural feed source for aquaculture or another purpose and needs further research.

Determining the nutrient removal capacity of duckweed Wolffia globosa under artificial conditions

Determining the nutrient removal capacity of duckweed Wolffia globosa under artificial conditions

The Dynamics of NO3- and NH4+ Uptake in Duckweed Are Coordinated with the Expression of Major Nitrogen Assimilation Genes.

Duckweed plants play important roles in aquatic ecosystems worldwide. They rapidly accumulate biomass and have potential uses in bioremediation of water polluted by fertilizer runoff or other chemicals. Here we studied the assimilation of two major sources of inorganic nitrogen, nitrate () and ammonium (), in six duckweed species: Spirodela polyrhiza, Landoltia punctata, Lemna aequinoctialis, Lemna turionifera, Lemna minor, and Wolffia globosa. All six duckweed species preferred over and started using only when was depleted. Using the available genome sequence, we analyzed the molecular structure and expression of eight key nitrogen assimilation genes in S. polyrhiza. The expression of genes encoding nitrate reductase and nitrite reductase increased about 10-fold when was supplied and decreased when was supplied. and induced the glutamine synthetase (GS) genes GS1;2 and the GS2 by 2- to 5-fold, respectively, but repressed GS1;1 and GS1;3. and upregulated the genes encoding ferredoxin- and NADH-dependent glutamate synthases (Fd-GOGAT and NADH-GOGAT). A survey of nitrogen assimilation gene promoters suggested complex regulation, with major roles for NRE-like and GAATC/GATTC cis-elements, TATA-based enhancers, repeats, and G-quadruplex structures. These results will inform efforts to improve bioremediation and nitrogen use efficiency.

Diet-induced Fasting Ghrelin Elevation Reflects the Recovery of Insulin Sensitivity and Visceral Adiposity Regression.

Lower fasting ghrelin levels (FGL) are associated with obesity and metabolic syndrome. We aimed to explore the dynamics of FGL during weight loss and its metabolic and adiposity-related manifestations beyond weight loss. This was a secondary analysis of a clinical trial that randomized participants with abdominal obesity/dyslipidemia to 1 of 3 diets: healthy dietary guidelines (HDG), Mediterranean diet (MED), or green-MED diet, all combined with physical activity (PA). Both MED diets were similarly hypocaloric and included 28 g/day walnuts. The green-MED group further consumed green tea (3-4 cups/day) and a Wolffia globosa (Mankai) plant green shake. We measured FGL and quantified body fat depots by magnetic resonance imaging at baseline and after 18 months. Among 294 participants (body mass index = 31.3 kg/m2; FGL = 504 ± 208 pg/mL; retention rate = 89.8%), lower FGL was associated with unfavorable cardiometabolic parameters such as higher visceral adipose tissue (VAT), intrahepatic fat, leptin, and blood pressure (P < 0.05 for all; multivariate models). The ∆FGL18-month differed between men (+7.3 ± 26.6%) and women (-9.2% ± 21.3%; P = 0.001). After 18 months of moderate and similar weight loss among the MED groups, FGL increased by 1.3%, 5.4%, and 10.5% in HDG, MED, and green-MED groups, respectively (P = 0.03 for green-MED vs HDG); sex-stratified analysis revealed similar changes in men only. Among men, FGL18-month elevation was associated with favorable changes in insulin resistance profile and VAT regression, after adjusting for relative weight loss (HbA1c: r = -0.216; homeostatic model of insulin resistance: r = -0.154; HDL-c: r = 0.147; VAT: r = -0.221; P < 0.05 for all). Insulin resistance and VAT remained inversely related with FGL elevation beyond that explained by weight loss (residual regression analyses; P < 0.05). Diet-induced FGL elevation may reflect insulin sensitivity recovery and VAT regression beyond weight loss, specifically among men. Green-MED diet is associated with greater FGL elevation.

Contribution to the floras of Lipetsk and Voronezh Regions (new vascular plant records from Usmansky pine forest)

The materials presented in the article supplement the information of “Flora of the middle zone of the European part of Russia” (Maevskiy, 2014) for the floras of Voronezh and Lipetsk Regions. The materials were collected in the period 2007–2020 on the protected areas of Usmansky Bor: in the Voronezhsky State Reserve, its protected zone and in the Federal Preserve “Voronezhsky”. The flora of the Lipetsk Region is supplemented by 9 species: Alchemilla argutiserrata, Aronia mitschurinii, Carex remota, C. tomentosa, Centaurea majorovii, Cotoneaster lucidus, Hieracium sylvularum, Scirpoides holoschoenus, Symphoricarpos albus. The data on 11 new taxa are given for the Voronezh Region flora: Cotoneaster lucidus, Crataegus ucrainica, Hieracium sylvularum, Nymphaea × marliacea, Phytolacca acinosa, Philadelphus latifolius, P. serotina, Spiraea chamaedryfolia, Thuja occidentalis, Veronica polita, Wolffia globosa. The article reports on new finds of Carex bohemica and Prunus cerasifera, species that were previously noted in the region, but not included in “Flora ...” (Maevskiy, 2014). Among the 20 marked species, 11 taxa are alien ornamental plants and fruit trees and shrubs noted outside of cultivation areas. Leaving the places of cultivation by alien plants and naturalization in natural communities is a modern problem of protected areas in the European part of Russia.

Measuring the effect of Mankai® (Wolffia globosa) on the gut microbiota and its metabolic output using an in vitro colon model

• Wolffia globosa an emerging functional food, targets obesity and metabolic disease. • Wolffia globosa rich in fiber and polyphenols modulates the gut microbiome. • Wolffia globosa increases microbiota phenolic acid and SCFA production in vitro. • Microbiome modulation may underpin certain health effects of Wolffia globosa ingestion. Mankai® is a cultivated strain of Wolffia globosa an aquatic plant of the family Lemnaceae commonly known as Duckweeds. Recent studies suggest that consumption of a Mankai® enriched diet may provide positive health effects by decreasing body weight and improving glucose homeostasis and plasma lipid profiles. However, the effects of Mankai® alone on the composition and metabolic output of the human gut microbiota has not been fully investigated. Here, Mankai® was digested and fermented in vitro using a batch culture model of the proximal colon. Inulin and cellulose were used as readily and poorly fermentable control fibers respectively. Mankai® significantly stimulated the production of phenolic metabolites and short chain fatty acids by the gut microbiota (p<0.05). Three major microbial metabolites, 3-4-hydroxyphenyl propionic acid, 3-3-hydroxyphenyl propanoic acid and protocatechuic acid were significantly increased after 24 h fermentation. Moreover, Mankai® treatment lowered the overall microbial diversity (p<0.05), in line with a selective microbiome modulation.