Inhibition of Fungal Strains Isolated from Cereal Grains via Vapor Phase of Essential Oils.

Fungal contamination in stored food grains is a global concern and affects food economics and human and animal health. It is clear that there is a need to develop new technologies with improved performances that are also eco-friendly in nature. Due to the bioactivity of essential oils (EOs) in the vapor phase, their low toxicity for humans, and their biodegradability and antifungal properties, EOs could be a suitable solution. In this study, we explored the potential of thyme, oregano, lemongrass, clove, and cajeput EOs in the vapor phase. For 17 days, inhibitory activity was assessed against five strains of postharvest pathogens-Aspergillus spp., Fusarium s. l. spp., and Penicilliumochrochloron-isolated from cereal grains. A modified disc volatilization method was used, which is more effective in comparison to traditional screening methods. Three concentrations were tested (250, 125, and 62.5 µL/L). The two highest concentrations resulted in complete inhibition of fungal growth; however, even 62.5 µL/L showed a significant antifungal effect. The efficiency of EOs followed this order: thyme > oregano > lemongrass > clove > cajeput. From our findings, it appears that the use of EOs vapors is a better option not only for laboratory experiments, but for subsequent practice.

Antifungal and synergistic activities of some selected essential oils on the growth of significant indoor fungi of the genus Aspergillus.

The study aimed to assess the antifungal activity of twenty-five essential oils (EOs) and the potential synergistic activity of the most effective EOs against significant indoor fungi of the genus Aspergillus [A. fumigatus (KBio-122), A. flavus (KBio-134), A. terreus (KBio-145) and A. niger (KBio-202)]. The chemical composition of all EOs was evaluated by the gas chromatography coupled with mass spectrometry (GC/MS) and gas chromatography with flame ionization detector (GC-FID) analysis. The antifungal susceptibility of EOs was evaluated by using the broth microdilution method. The most effective EOs were selected to determine the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) at a concentration range from 256 to 0.125 µg/mL. For the synergistic activities, the most effective EOs were tested using the chessboard pattern. The most sensitive strain to treatments with essential oils alone and in the combination of EOs was A. flavus (KBio-134). The chessboard assay showed that combinations of lemongrass and thyme EOs proved the most potent synergistic antifungal activity (FICI = 0.1875) against A. fumigatus (KBio-122). The synergy displayed by a combination of some EOs may be used to control fungal growth or increasing resistance to available synthetic antifungals, consequently permitting the reduction of their most active doses.

"Salicylic Acid Mutant Collection" as a Tool to Explore the Role of Salicylic Acid in Regulation of Plant Growth under a Changing Environment.

The phytohormone salicylic acid (SA) has a crucial role in plant physiology. Its role is best described in the context of plant response to pathogen attack. During infection, SA is rapidly accumulated throughout the green tissues and is important for both local and systemic defences. However, some genetic/metabolic variations can also result in SA overaccumulation in plants, even in basal conditions. To date, more than forty Arabidopsis thaliana mutants have been described as having enhanced endogenous SA levels or constitutively activated SA signalling pathways. In this study, we established a collection of mutants containing different SA levels due to diverse genetic modifications and distinct gene functions. We chose prototypic SA-overaccumulators (SA-OAs), such as bon1-1, but also "non-typical" ones such as exo70b1-1; the selection of OA is accompanied by their crosses with SA-deficient lines. Here, we extensively studied the plant development and SA level/signalling under various growth conditions in soil and in vitro, and showed a strong negative correlation between rosette size, SA content and PR1/ICS1 transcript signature. SA-OAs (namely cpr5, acd6, bon1-1, fah1/fah2 and pi4kß1ß2) had bigger rosettes under high light conditions, whereas WT plants did not. Our data provide new insights clarifying a link between SA and plant behaviour under environmental stresses. The presented SA mutant collection is thus a suitable tool to shed light on the mechanisms underlying trade-offs between growth and defence in plants.

Metabolism of Stilbenoids by Human Faecal Microbiota.

Stilbenoids are dietary phenolics with notable biological effects on humans. Epidemiological, clinical, and nutritional studies from recent years have confirmed the significant biological effects of stilbenoids, such as oxidative stress protection and the prevention of degenerative diseases, including cancer, cardiovascular diseases, and neurodegenerative diseases. Stilbenoids are intensively metabolically transformed by colon microbiota, and their corresponding metabolites might show different or stronger biological activity than their parent molecules. The aim of the present study was to determine the metabolism of six stilbenoids (resveratrol, oxyresveratrol, piceatannol, thunalbene, batatasin III, and pinostilbene), mediated by colon microbiota. Stilbenoids were fermented in an in vitro faecal fermentation system using fresh faeces from five different donors as an inoculum. The samples of metabolized stilbenoids were collected at 0, 2, 4, 8, 24, and 48 h. Significant differences in the microbial transformation among stilbene derivatives were observed by liquid chromatography mass spectrometry (LC/MS). Four stilbenoids (resveratrol, oxyresveratrol, piceatannol and thunalbene) were metabolically transformed by double bond reduction, dihydroxylation, and demethylation, while batatasin III and pinostilbene were stable under conditions simulating the colon environment. Strong inter-individual differences in speed, intensity, and pathways of metabolism were observed among the faecal samples obtained from the donors.

Polysaccharides from Basidiocarps of Cultivating Mushroom Pleurotus ostreatus: Isolation and Structural Characterization.

Oyster mushrooms are an interesting source of biologically active glucans and other polysaccharides. This work is devoted to the isolation and structural characterization of polysaccharides from basidiocarps of the cultivated oyster mushroom, Pleurotus ostreatus. Five polysaccharidic fractions were obtained by subsequent extraction with cold water, hot water and two subsequent extractions with 1 m sodium hydroxide. Branched partially methoxylated mannogalactan and slightly branched (1→6)-ß-d-glucan predominated in cold- and hot-water-soluble fractions, respectively. Alternatively, these polysaccharides were obtained by only hot water extraction and subsequent two-stage chromatographic separation. The alkali-soluble parts originating from the first alkali extraction were then fractionated by dissolution in dimethyl sulfoxide (DMSO). The polysaccharide insoluble in DMSO was identified as linear (1→3)-α-d-glucan, while branched (1→3)(1→6)-ß-d-glucans were found to be soluble in DMSO. The second alkaline extract contained the mentioned branched ß-d-glucan together with some proteins. Finally, the alkali insoluble part was a cell wall complex of chitin and ß-d-glucans.

Effect of Selected Stilbenoids on Human Fecal Microbiota.

Dietary phenolics or polyphenols are mostly metabolized by the human gut microbiota. These metabolites appear to confer the beneficial health effects attributed to phenolics. Microbial composition affects the type of metabolites produced. Reciprocally, phenolics modulate microbial composition. Understanding this relationship could be used to positively impact health by phenolic supplementation and thus create favorable colonic conditions. This study explored the effect of six stilbenoids (batatasin III, oxyresveratrol, piceatannol, pinostilbene, resveratrol, thunalbene) on the gut microbiota composition. Stilbenoids were anaerobically fermented with fecal bacteria from four donors, samples were collected at 0 and 24 h, and effects on the microbiota were assessed by 16S rRNA gene sequencing. Statistical tests identified affected microbes at three taxonomic levels. Observed microbial composition modulation by stilbenoids included a decrease in the Firmicutes to Bacteroidetes ratio, a decrease in the relative abundance of strains from the genus Clostridium, and effects on the family Lachnospiraceae. A frequently observed effect was a further decrease of the relative abundance when compared to the control. An opposite effect to the control was observed for Faecalibacterium prausnitzii, whose relative abundance increased. Observed effects were more frequently attributed to resveratrol and piceatannol, followed by thunalbene and batatasin III.

The effect of oregano essential oil on microbial load and sensory attributes of dried meat.

BACKGROUND: Microbial load can be controlled using either synthetic or natural preservatives. Particular interest has been focused on the potential application of plant essential oils as safer additives for meat. However, there is no published research on the use of essential oils during the meat drying process. This study was focused on enhancing the meat drying process by using oregano essential oil (OEO) to inhibit the growth of bacteria and thus obtain a value-added dried meat product. The sensory response from assessors is presented. RESULTS: It was found that the application of OEO in meat was effective in inhibiting Salmonella enteritidis and Escherichia coli. After 6 h of drying at 55 °C, 2 mL (0.038 mL L-1 air) and 1.5 mL (0.028 mL L-1 air) of OEO were considered as the minimal inhibitory concentrations (MICs) against S. enteritidis and E. coli respectively. Samples treated with 0.75 mL of OEO were more attractive for consumption compared with the control; at a higher concentration of OEO, the sensory quality of the food was affected. CONCLUSION: A value-added dried meat product obtained by using OEO to enhance food safety received an acceptable sensory response from consumers. © 2016 Society of Chemical Industry.

Antifungal effect of essential oil components against Aspergillus niger when loaded into silica mesoporous supports.

BACKGROUND: Essential oil components (EOCs) are known for their antifungal properties; however, their high volatility limits their application as antimicrobial agents. Strategies used for controlling the volatility of EOCs include encapsulation or loading into porous materials. This study evaluated the in vitro antifungal activity of selected EOCs (carvacrol, cinnamaldehyde, eugenol and thymol) against the fungus Aspergillus niger when loaded into MCM-41 and ß-cyclodextrin (ß-CD). RESULTS: Carvacrol and thymol in Mobil Composition of Matter No. 41 (MCM-41) displayed remarkable enhanced antifungal properties in comparison to the pure or ß-CD-encapsulated EOCs. In fact, carvacrol and thymol were able to maintain antifungal activity and inhibit fungal growth for 30 days, suggesting better applicability of these EOCs as natural preservatives. CONCLUSIONS: The sustained antifungal effect of EOCs encapsulated into silica mesoporous supports was described.

Antibacterial activities of plant-derived compounds and essential oils toward Cronobacter sakazakii and Cronobacter malonaticus.

Cronobacter sakazakii and C. malonaticus are opportunistic pathogens that cause infections in children and immunocompromised adults. In the present study, the antibacterial activity of 19 plant-derived compounds, 5 essential oils, and an extract of propolis were assessed against C. sakazakii and C. malonaticus. The effects of most of these antimicrobials have not been reported previously. Both strains were susceptible to thymol, carvacrol, thymoquinone, p-cymene, linalool, camphor, citral, eugenol, and trans-cinnamaldehyde as well as cinnamon, lemongrass, oregano, clove, and laurel essential oils; their minimum inhibitory concentrations varied between 0.1 and 2.0 mg/mL. As an alternative treatment method, vapors of the volatiles were tested as an indirect treatment. Vapors of trans-cinnamaldehyde, eugenol, oregano, and cinnamon essential oils inhibited both tested strains, while vapors of linalool were only active against C. sakazakii. To our knowledge, this study is the first time that the inhibitory activity of the vapors of these compounds and essential oils has been reported against Cronobacter spp.

Consumer acceptance and quality of game meat "droëwors" sausages with different levels of added fat.

Droëwors (dried sausage) is a unique dried meat product from South Africa, which is not smoked, fermented, nor is nitrite used in its production. The objective of the study was to compare the quality parameters and consumer acceptance of common eland meat droëwors with different quantities of added beef fat. Three treatments containing either 10, 15, or 20% (by weight) added beef fat were compared regarding chemical composition, fatty acid profiles, and sensory properties. Increasing the amount of fat in the dried product resulted in a significant decrease in the protein and ash percentages. The sausages with 10% added fat had the most favourable fatty acid profile in terms of nutritional value. Several differences were found in the sensory profiles of the products evaluated by the trained sensory panel; in particular, the evaluation scores for texture improved with increasing fat content. No differences were found between the products by the untrained consumer panel for appearance and taste. When the dataset was analysed according to gender, it was found that women evaluated the 10% fat samples more favourably. This is in line with their stated preference for low-fat meat products. For these reasons, the production of sausages with the lowest fat content can be recommended as they best meet consumer expectations.

The Effect of Carbamazepine on Performance, Carcass Value, Hematological and Biochemical Blood Parameters, and Detection of Carbamazepine and Its Metabolites in Tissues, Internal Organs, and Body Fluids in Growing Rabbits.

Antiepileptic drugs (e.g., carbamazepine; CBZ) are widely prescribed for various conditions beyond epilepsy, including neurologic and psychiatric disorders. These medications can have both favorable and unfavorable impacts on mood, anxiety, depression, and psychosis. CBZ has been found at low concentrations (in the unit of nanograms per liter) in rivers, surface water, and even drinking water. As a result, when reclaimed wastewater is used for irrigation in agricultural ecosystems, CBZ can be reintroduced into the environment. That is why we tested different doses of CBZ in rabbits' feed as the meat is consumed in every community, has no religious barriers, and the potential risk of consuming meat which has been exposed to CBZ treatment is not known. Also, the evidence of the effect of CBZ on rabbits is missing. Mainly, the CBZ doses affected the count of leukocytes and other blood traits, meaning the higher the dose, the higher the reduction. Moreover, there were only low amounts of CBZ in rabbits' meat or tissues when they were exposed to the treatment.

Effect of Conventional Preservatives and Essential Oils on the Survival and Growth of Escherichia coli in Vegetable Sauces: A Comparative Study.

Essential oils have gained attention as natural alternatives to chemical preservatives in food preservation. However, more information is needed regarding consumer acceptance of essential oils in actual food products. This study aimed to compare the effects of conventional preservatives, heat treatment, and essential oils derived from thyme, oregano, and lemongrass on the survival and growth of pathogenic Escherichia coli in vegetable sauces. The results demonstrated a gradual decrease in pathogen numbers over time, even in untreated samples. On the fifth day of storage, heat treatment, sodium chloride, and acidification using citric acid (pH 3.2) exhibited reductions of 4.4 to 5.3 log CFU/g compared to the untreated control. Among the essential oils tested, lemongrass essential oil at a concentration of 512 mg/kg demonstrated the most remarkable effectiveness, resulting in a reduction of 1.9 log CFU/g compared to the control. Fifteen days after treatment, the control samples exhibited a contamination rate of 6.2 log CFU/g, while E. coli numbers in treated samples with heat, sodium chloride, citric acid (pH 3.2), and lemongrass essential oil (512 mg/kg) were below the detection limits. Additionally, sensory evaluation was conducted to assess the acceptability of the treated samples. The findings provide valuable insights into the potential utilisation of essential oils as natural preservatives in vegetable sauces and their impact on consumer acceptance.

Mid-Infrared Spectroscopic Study of Cultivating Medicinal Fungi Ganoderma: Composition, Development, and Strain Variability of Basidiocarps.

Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy was proposed for rapid, versatile, and non-invasive screening of Ganoderma basidiocarps to assess their potential for specific applications. Fifteen species and strains of this fungus were selected for analysis, and fine sections at different parts of young and mature basidiocarps were obtained. The spectra of fungal samples showed significant differences interpreted in terms of biochemical composition using characteristic bands of proteins, polysaccharides, lipids, and triterpenoids. Obviously, for the transverse sections in trama, especially in the basal part, the most intense bands at 950-1200 cm-1 corresponded to polysaccharide vibrations, while for the superficial sections, the bands of carbonyl and aliphatic groups of triterpenoids at 1310-1470, 1550-1740, and 2850-2980 cm-1 predominated. The pilei, especially hymenium tubes, apparently contained more proteins than the bases and stipes, as evidenced by the intense bands of amide vibrations at 1648 and 1545-1550 cm-1. The specificity of the Ganoderma basidiocarp is a densely pigmented surface layer rich in triterpenoids, as proved by ATR-FTIR spectroscopy. The spectral differences corresponding to the specificity of the triterpenoid composition may indicate the prospects of individual strains and species of this genus for cultivation and further use in food, cosmetics, or medicine.

Phytocannabinoid-rich galenic preparations for topical administration: extraction and stability testing.

Although medical cannabis was legalized in Czechia in 2013 and its use in topical treatments of skin disorders is now allowed, galenic formulations prepared from medical cannabis have not been widely implemented in the Czech healthcare system. One of the main reasons is the lack of a straightforward standardized protocol for their preparation. Cannabinoids, e.g., cannabidiol (CBD) and tetrahydrocannabinol (THC), have been shown to have therapeutic effects on various skin conditions, such as atopic dermatitis, psoriasis, scleroderma, acne and skin pigmentation. Recognizing the potential of dermatological treatment with medical cannabis, the present study aimed to evaluate the extraction capacity of various pharmaceutical bases for cannabinoids and the stability of prepared galenic formulations for dermatological applications with respect to cannabinoid content. The results showed that the stability of cannabinoids in formulations depended on the bases' physical and chemical properties. The highest THC decomposition was observed in cream bases and Vaseline, with estimated percentage loss of total content of up to 5.4% and 5.6% per week, respectively. In contrast, CBD was more stable than THC. Overall, the tested bases were comparably effective in extracting cannabinoids from plant material. However, olive oil and Synderman bases exhibited the highest cannabinoid extraction efficiencies (approximately 70%) and the best storage stabilities in terms of the content of monitored compounds. The proposed preparation protocol is fast and easily implementable in pharmacies and medical facilities.

Biochemical and physiological changes in Zea mays L. after exposure to the environmental pharmaceutical pollutant carbamazepine.

The presence of pharmaceuticals in the environment is a matter of great concern. They are consistently found in the environment, raising concerns regarding human exposure through dietary intake. In this study, we observed the effect of the application of carbamazepine at 0.1, 1, 10, and 1000 µg per kg of soil contamination levels to assess stress metabolism in Zea mays L. cv. Ronaldinio at the 4th leaf, tasselling, and dent phenological stages. The transfer of carbamazepine to the aboveground and root biomass was assessed, and uptake increased dose-dependently. No direct effect on biomass production was observed, but multiple physiological and chemical changes were observed. Major effects were consistently observed at the 4th leaf phenological stage for all contamination levels, including reduced photosynthetic rate, reduced maximal and potential activity of photosystem II, decreased water potential, decreased carbohydrates (glucose and fructose) and γ-aminobutyric acid in roots, and increased maleic acid and phenylpropanoids (chlorogenic acid and its isomer, 5-O-caffeoylquinic acid) in aboveground biomass. A reduction in net photosynthesis was observed for the older phenological stages, whereas no other relevant and consistent physiological and metabolic changes related to contamination exposure were detected. Our results indicate that Z. mays can overcome the environmental stress caused by the accumulation of carbamazepine with notable metabolic changes at the early phenological stage; however, older plants adapted and only exhibited minor effects in the presence of the contaminant. The potential implications for agricultural practice could be associated with the plant's response to simultaneous stresses due to metabolite changes associated with oxidative stress.

Contaminants and their ecological risk assessment in beach sediments and water along the Maharashtra coast of India: A comprehensive approach using microplastics, heavy metal(loid)s, pharmaceuticals, personal care products and plasticisers.

Emerging contaminants and their pervasive presence in freshwater ecosystems have been widely documented, but less is known about their prevalence and the harm they cause in marine ecosystems, particularly in developing countries. This study provides data on the prevalence and risk posed by microplastics, plasticisers, pharmaceuticals and personal care products (PPCPs), and heavy metal(loid)s (HMs) along the Maharashtra coast of India. The sediment and coastal water samples were collected from 17 sampling stations, processed, and subjected to FTIR-ATR, ICP-MS, SEM-EDX, LC-MS/MS, and GC-MS for further analysis. Higher MPs abundance, combined with the pollution load index, indicates that the northern zone is a high-impact zone with pollution concerns. Plasticisers in extracted MPs and HMs adsorption on MPs surface from surrounding waters reveal their roles as a source and vector for contaminants, respectively. The mean concentration of metoprolol (53.7-306 ng L-1), tramadol (16.6-198 ng L-1), venlafaxine (24.6-234 ng L-1), and triclosan (211-433 ng L-1) in Maharashtra's coastal waters were several folds higher than in other water systems, raising major health concerns. The hazard quotient (HQ) scores revealed that >70 % of study sites pose a high to medium (1 > HQ > 0.1) ecological risk to fish, crustaceans and algae, indicating serious concern. Fish and crustaceans (35.3 % each) show a higher level of risk than algae (29.5 %). Metoprolol and venlafaxine could represent greater ecological risks than tramadol. Similarly, HQ suggests that bisphenol A has larger ecological risks than bisphenol S along the Maharashtra coast. To the best of our knowledge, this is the first in-depth investigation into emerging pollutants in Indian coastal regions. This information is crucial for better policy formulation and coastal management in India in general, and Maharashtra in particular.

Essential oils in the ranunculaceae family: chemical composition of hydrodistilled oils from Consolida regalis, Delphinium elatum, Nigella hispanica, and N. nigellastrum seeds.

In this study, we analyzed the chemical composition of volatile oils hydrodistilled from seeds of Consolida regalis, Delphinium elatum, Nigella hispanica, and N. nigellastrum using GC and GC/MS. In C. regalis, octadecenoic (77.79%) and hexadecanoic acid (8.34%) were the main constituents. Similarly, the oils from D. elatum and N. hispanica seeds consisted chiefly of octadecadienoic (42.83 and 35.58%, resp.), hexadecanoic (23.87 and 28.59%, resp.), and octadecenoic acid (21.67 and 19.76%, resp.). Contrastingly, the monoterpene hydrocarbons α-pinene (34.67%) and ß-pinene (36.42%) were the main components of N. nigellastrum essential oil. Our results confirm the presence of essential oils in the family Ranunculaceae and suggest chemotaxonomical relationships within the representatives of the genera Consolida, Delphinium, and Nigella. In addition, the presence of various bioactive constituents such as linoleic acid, (-)-ß-pinene, squalene, or carotol in seeds of D. elatum, N. hispanica, and N. nigellastrum indicates a possible industrial use of these plants.

Attraction or Repelling Effects of Commercial Plant Essential Oils on the Synanthropic Cheiracanthium mildei (Araneae: Cheiracanthiidae).

The northern yellow sac spider Cheiracanthium mildei L. Koch, is expanding its range to Central Europe, especially to synanthropic habitats. The spiders become unwanted companions because of the unreasonable fear - arachnophobia, and estetic reason - silk retreats in corners, capturing dust. The most commonly used substances against spiders are pesticides, which are, however, toxic. In our work we tested the attraction or repellence of 15 essential oils (EO) from plants representing eight families to C. mildei. Our research has shown a significant repellent effect of EO from three plants, namely Syzygium aromaticum (L.) Merr. et L. M. Perry (Myrtales: Myrtaceae), Ananas comosus (L.) Merr. (Poales: Bromeliaceae) and Musa sp. (L.) (Zingiberales: Musaceae). In contrast, some EOs appeared to have an attraction effect, particularly Carum carvi L. (Apiales: Apiaceae). Zingiber officinale Roscoe (Zingiberales: Zingiberaceae) reduced the tendency of spiders to construct the silken retreat. S. aromaticum, A. sativus, Musa sp. and Z. officinale have the potential to be used as natural repellents against spiders.

Amino Acid Supplementation as a Biostimulant in Medical Cannabis (Cannabis sativa L.) Plant Nutrition.

There is growing evidence to support the involvement of nutrients and biostimulants in plant secondary metabolism. Therefore, this study evaluated the potential of amino acid-based supplements that can influence different hydroponic nutrient cycles (systems) to enhance the cannabinoid and terpene profiles of medical cannabis plants. The results demonstrate that amino acid biostimulation significantly affected ion levels in different plant tissues (the "ionome"), increasing nitrogen and sulfur content but reducing calcium and iron content in both nutrient cycles. A significantly higher accumulation of nitrogen and sulfur was observed during the recirculation cycle, but the calcium level was lower in the whole plant. Medical cannabis plants in the drain-to-waste cycle matured 4 weeks earlier, but at the expense of a 196% lower maximum tetrahydrocannabinolic acid yield from flowers and a significantly lower concentration of monoterpene compounds than in the recirculation cycle. The amino acid treatments reduced the cannabinolic acid content in flowers by 44% compared to control in both nutritional cycles and increased the monoterpene content (limonene) up to 81% in the recirculation cycle and up to 123% in the drain-to-waste cycle; ß-myrcene content was increased up to 139% in the recirculation cycle and up to 167% in the drain-to-waste cycle. Our results suggest that amino acid biostimulant supplements may help standardize the content of secondary metabolites in medical cannabis. Further experiments are needed to identify the optimal nutrient dosage and method of administration for various cannabis chemotypes grown in different media.

Polysaccharides from Basidiocarps of the Polypore Fungus Ganoderma resinaceum: Isolation and Structure.

In this study, we focused on the isolation and structural characterization of polysaccharides from a basidiocarp of polypore fungus Ganoderma resinaceum. Polysaccharide fractions were obtained by successive extractions with cold water at room temperature (20 °C), hot water under reflux (100 °C), and a solution of 1 mol L-1 sodium hydroxide. The purity of all fractions was controlled mainly by Fourier transform infrared (FTIR) spectroscopy, and their composition and structure were characterized by organic elemental analysis; neutral sugar and methylation analyses by gas chromatography equipped with flame ionization detector (GC/FID) and mass spectrometry detector (GC/MS), respectively; and by correlation nuclear magnetic resonance (NMR) spectroscopy. The aqueous extracts contained two main polysaccharides identified as a branched O-2-ß-d-mannosyl-(1→6)-α-d-galactan and a highly branched (1→3)(1→4)(1→6)-ß-d-glucan. Mannogalactan predominated in the cold water extract, and ß-d-glucan was the main product of the hot water extract. The hot water soluble fraction was further separated by preparative anion exchange chromatography into three sub-fractions; two of them were identified as branched ß-d-glucans with a structure similar to the corresponding polysaccharide of the original fraction. The alkaline extract contained a linear (1→3)-α-d-glucan and a weakly branched (1→3)-ß-d-glucan having terminal ß-d-glucosyl residues attached to O-6 of the backbone. The insoluble part after all extractions was identified as a polysaccharide complex containing chitin and ß-d-glucans.