Chemical Composition and Anti-Inflammatory and Antioxidant Activities of Extracts from Cultivated Morel Mushrooms, Species of Genus Morchella (Ascomycota).

Species of genus Morchella are high-value edible mushrooms. They are sought after by culinary experts due to their aroma, flavor, meaty texture, and health benefits. M. rufobrunnea, M. sextelata, and M. americana were chosen in this study and investigated for their medicinal quality by using in vitro anti-inflammatory and antioxidant assays. This sampling represents conditions by which morels are produced (cultivated indoors, cultivated outdoors, and collected from natural habitats, respectively) for commercial markets. Both aqueous and methanolic extracts of all three morel species showed identical chromatographic and bioassay profiles, independent of their phylogenetic position or production method. In an antioxidant assay, aqueous and methanolic extracts of these mushrooms at 100 µg/mL inhibited lipid peroxidation (LPO) by 59%-62% and 33%-36%, respectively. In an anti-inflammatory assay using cyclooxygenase enzymes (COX-1 and COX-2), aqueous and methanolic extracts at 100 µg/mL showed COX-1 enzyme inhibition by 53%-57% and 30%-32% and COX-2 enzyme inhibition by 38%-44% and 16%-17%, respectively. Chromatographic purification and spectroscopic characterization of M. rufobrunnea extracts afforded five sugars (compounds 1-5), seven organic acids (compounds 6-13), three flavonoids (compounds 14-16), triglycerides, free fatty acids, and three sterols (compounds 17-19). This is the first report of COX-1 and COX-2 enzymes and LPO inhibitory activities of pure isolates (S)-morelid (compound 6), glutamic acid (compound 9), and brassicasterol (compound 19). This study also showed inhibitions of COX-1 (by 84%, 33%, and 37%), COX-2 (by 47%, 11%, and 22%), and LPO (by 74%, 48%, and 35%), respectively, at 25 µg/mL.

Mercury speciation, bioavailability, and biomagnification in contaminated streams on the Savannah River Site (SC, USA).

Water, sediment, and biota from two streams on the Savannah River Site were sampled to study mercury (Hg) biogeochemistry. Total and methyl- Hg (MHg) concentrations were measured for all samples, speciation models were used to explore Hg speciation in the water, and Diffusive gradients in thin films (DGT) were applied to indicate the vertical profiles of labile Hg (DGT-Hg). Trophic position (δ15N) was estimated for biota and used to establish MHg biomagnification model. The speciation model indicated Hg methylation in the water occurred on settling particles and the most bioavailable Hg species to bacteria were complexes of inorganic Hg and labile organic ligands. Correspondingly, dissolved organic carbon concentrations were positively related to MHg concentrations in the water. In the sediment, the sharp increase of DGT-Hg around the sediment water interface underscores the importance of this interface, which determines the differences in the accumulation and generation of labile Hg among different waterbodies. The positive correlation between sediment MHg and sulfate concentrations suggested possible methylation reaction by dissimilatory sulfate reducing bacteria in the sediment. The food web magnification factors of MHg were 9.6 (95% CI: 4.0-23.4) and 4.4 (95% CI: 2.5-7.7) for the two streams established with trophic data of biofilm, invertebrates, and fish. Meanwhile, DGT-Hg concentrations in the water were positively correlated to biofilm Hg concentrations, which can be combined with the MHg biomagnification model to generate a modified biomagnification model that estimate MHg bioaccumulation with only labile Hg concentrations in the water. With this approach, Hg accumulation in abiotic and biotic environmental compartments was connected and the different bioaccumulation patterns of Hg in different waterbodies were explained with both geochemical and biological factors.

Evaluation of the DGT technique for predicting uptake of metal mixtures by fathead minnow (Pimephales promelas) and yellow lampmussel (Lampsilis cariosa).

Diffusive gradients in thin films (DGT) were assessed for their predictive capability of fathead minnow and yellow lampmussel bioaccumulation in copper (Cu) and lead (Pb) mixed metal exposures. Nine treatments with a matrix of 3 Cu and 3 Pb concentrations were utilized. Exposures were coupled, with organisms and DGT exposed in tanks for 6 days. The Cu measured in fish, mussel, and DGT was found not to be influenced by Pb treatment, whereas Pb accumulation was impacted by the interaction of Cu and Pb treatment. The Pb accumulation increased with increasing Cu concentration, which was attributed to the different speciation of Cu and Pb in the water where Cu binds preferentially to ligands, decreasing its bioavailability and concomitantly displacing Pb from complexing sites. The DGT values were significantly correlated with accumulated Cu and Pb in the fish, but not with Pb in the mussel. In addition, DGT was determined to better predict aquatic organism bioaccumulation of Cu than the inorganic Cu fraction calculated by the speciation model, because DGT accumulated not only inorganic metal fractions but also complexes of metal and organic matter. The present study provides insights into metal speciation in polluted environments, extends the understanding of using DGT as a tool for estimating metal bioavailability, and provides implications for the selection of geochemical modeling, biological sampling, and passive sampling techniques for monitoring trace metal contamination. Environ Toxicol Chem 2019;38:61-70. © 2018 SETAC.

Impact of natural organic matter and increased water hardness on DGT prediction of copper bioaccumulation by yellow lampmussel (Lampsilis cariosa) and fathead minnow (Pimephales promelas).

We conducted an exposure experiment with Diffusive Gradients in Thin- Films (DGT), fathead minnow (Pimephales promelas), and yellow lampmussel (Lampsilis cariosa) to estimate bioavailability and bioaccumulation of Cu. We hypothesized that Cu concentrations measured by DGT can be used to predict Cu accumulation in aquatic animals and alterations of water chemistry can affect DGT's predict ability. Three water chemistries (control soft water, hard water, and addition of natural organic matter (NOM)) and three Cu concentrations (0, 30, and 60 µg/L) were selected, so nine Cu-water chemistry combinations were used. NOM addition treatments resulted in decreased concentrations of DGT-measured Cu and free Cu ion predicted by Biotic Ligand Model (BLM). Both hard water and NOM addition treatments had reduced concentrations of Cu ion and Cu-dissolved organic matter complexes compared to other treatments. DGT-measured Cu concentrations were linearly correlated to fish accumulated Cu, but not to mussel accumulated Cu. Concentrations of bioavailable Cu predicted by BLM, the species complexed with biotic ligands of aquatic organisms and, was highly correlated to DGT-measured Cu. In general, DGT-measured Cu fit Cu accumulations in fish, and this passive sampling technique is acceptable at predicting Cu concentrations in fish in waters with low NOM concentrations.

Do constructed wetlands remove metals or increase metal bioavailability?

The H-02 wetland was constructed to treat building process water and storm runoff water from the Tritium Processing Facility on the Department of Energy's Savannah River Site (Aiken, SC). Monthly monitoring of copper (Cu) and zinc (Zn) concentrations and water quality parameters in surface waters continued from 2014 to 2016. Metal speciation was modeled at each sampling occasion. Total Cu and Zn concentrations released to the effluent stream were below the NPDES limit, and the average removal efficiency was 65.9% for Cu and 71.1% for Zn. The metal-removal processes were found out to be seasonally regulated by sulfur cycling indicated by laboratory and model results. High temperature, adequate labile organic matter, and anaerobic conditions during the warm months (February to August) favored sulfate reduction that produced sulfide minerals to significantly remove metals. However, the dominant reaction in sulfur cycling shifted to sulfide oxidation during the cool months (September to next March). High concentrations of metal-organic complexes were observed, especially colloidal complexes of metal and fulvic acid (FA), demonstrating adsorption to organic matter became the primary process for metal removal. Meanwhile, the accumulation of metal-FA complexes in the wetland system will cause negative effects to the surrounding environment as they are biologically reactive, highly bioavailable, and can be easily taken up and transferred to ecosystems by trophic exchange.

Evaluation of diffusive gradients in thin films for prediction of copper bioaccumulation by yellow lampmussel (Lampsilis cariosa) and fathead minnow (Pimephales promelas).

Using a coupled method of diffusive gradients in thin films (DGT) exposure with aquatic organism bioassays, we assessed the use of DGT as a tool for estimating copper (Cu) bioavailability in contaminated waters. The DGT-accumulated Cu fraction could possibly be used as a surrogate for other assessments of metal bioavailability. The Cu concentrations in fathead minnow (Pimephales promelas) and yellow lampmussel (Lampsilis cariosa) soft tissue were compared with DGT-accumulated Cu after 2, 4, and 6 d of exposure to a Cu concentration series in static, water-only assays. The DGT-accumulated Cu was found to include free Cu ions, labile inorganic Cu complexes, and labile dissolved organic matter Cu complexes, compared with Cu speciation output from the biotic ligand model. Regressions of Cu concentrations between DGT and fathead minnow at 4 and 6 d of exposure demonstrated linear relationships. The Cu bioaccumulated in yellow lampmussel was overpredicted by DGT at Cu concentrations greater than 10 µg L-1 , which may be caused by internal regulation of Cu. The speciation component of the biotic ligand model predicted relationships between inorganic Cu and animal-accumulated Cu that were similar to predicted relationships between DGT-indicated Cu and animal-accumulated Cu at all deployment durations. Environ Toxicol Chem 2018;37:1535-1544. © 2018 SETAC.

Exposure to mercury and Aroclor 1268 congeners in least terns (Sternula antillarum) in coastal Georgia, USA.

Concentrations of total mercury and the rare PCB mixture Aroclor 1268 in least terns (Sternula antillarum), a colonially-nesting, piscivorous seabird, in the Turtle River estuary and other coastal sites in Georgia, USA, were investigated. The Turtle River estuary is the location of the Linden Chemical Plant (LCP) Superfund site, a site prioritized by USA law for immediate remediation, where industries released effluent containing these contaminants until 1994. Aroclor 1268 is a highly-chlorinated PCB mixture that was used and released exclusively at the LCP site and nowhere else in the south eastern USA. High concentrations of Aroclor 1268 congeners and mercury have been documented in biota local to LCP, but no studies report concentrations in high trophic level, piscivorous birds such as least terns. We collected feathers and feces from chicks, and eggs from adults, at nesting colonies along the Georgia coast to analyze contaminant loads (in dry weight ppb). Mean Aroclor 1268 mixture concentrations in eggs (≤16,329 ppb) were highest at colonies in and just outside LCP, and decreased with increasing distance (up to 110 km) from LCP, but the Aroclor 1268 signature congener mixture was present at all sites. Mercury concentrations in eggs (≤3370 ppb), feathers (≤5950 ppb), and feces (≤417 ppb), were present at all sites, but did not vary significantly among sites. This report confirms the extensive dispersal of Aroclor 1268 congeners (approximately 110 km north and 70 km south of its point source) via bioaccumulation and trophic transfer.

Testing for associations between hematozoa infection and mercury in wading bird nestlings.

Several wading bird species in the southeastern US have a history of infection by hematozoa/avian malaria as well as mercury accumulation through their diet, and thus may be exposed to two, generally sublethal, yet chronic, stressors. We analyzed nestling wading birds (n = 171) of varying size and trophic position from the southeastern US, and a smaller sample (n = 23) of older, free-ranging birds, to look for potential interrelationships between infection by hematozoa and mercury (Hg) uptake. Only one nestling was PCR positive for hematozoa (Plasmodium/Haemoproteus) whereas nine (39%) of the older wading birds were positive. Sequencing indicated that both nestling and adult positives were infected with Plasmodium. Given the low infection rate of the nestlings, there was no association between Hg and malaria. The older birds exhibited a possible malaria/Hg association, but it may be confounded by their greater potential exposure period and large-scale movements.

Precipitation influences on uptake of a global pollutant by a coastal avian species.

Climatic variation, including precipitation amounts and timing, has been linked to abundance and breeding success of many avian species. Less studied, but also of significance, is the consequence of climatic variability on the exposure and uptake of nutrients and contaminants by wildlife. The authors examined mercury (Hg) concentrations in nestling wood stork feathers in a coastal setting over a 16-yr period to understand the influence of rainfall amounts on Hg transfer by parental provisioning relative to habitat use, assuming differential bioavailability of Hg within freshwater and saltwater habitat types. Coastal Hg uptake by stork nestlings was linked to freshwater habitat use, as indicated by stable carbon isotope (δ(13)C) analyses. Cumulative rainfall amounts exceeding 220 cm in the 23 mo preceding the breeding seasons resulted in greater use of freshwater wetlands as foraging habitat and greater Hg accumulation by nestling storks.

Spatial and taxonomic variation in trace element bioaccumulation in two herbivores from a coal combustion waste contaminated stream.

Dissimilarities in habitat use, feeding habits, life histories, and physiology can result in syntopic aquatic taxa of similar trophic position bioaccumulating trace elements in vastly different patterns. We compared bioaccumulation in a clam, Corbicula fluminea and mayfly nymph Maccaffertium modestum from a coal combustion waste contaminated stream. Collection sites differed in distance to contaminant sources, incision, floodplain activity, and sources of flood event water and organic matter. Contaminants variably accumulated in both sediment and biofilm. Bioaccumulation differed between species and sites with C. fluminea accumulating higher concentrations of Hg, Cs, Sr, Se, As, Be, and Cu, but M. modestum higher Pb and V. Stable isotope analyses suggested both spatial and taxonomic differences in resource use with greater variability and overlap between species in the more physically disturbed site. The complex but essential interactions between organismal biology, divergence in resource use, and bioaccumulation as related to stream habitat requires further studies essential to understand impacts of metal pollution on stream systems.

Variation in trace-element accumulation in predatory fishes from a stream contaminated by coal combustion waste.

Extensive and critical evaluation can be required to assess contaminant bioaccumulation in large predatory fishes. Species differences in habitat use, resource use, and trophic level, often influenced by body form, can result in diverging contaminant bioaccumulation patterns. Moreover, the broad size ranges inherent with large-bodied fish provide opportunity for trophic and habitat shifts within species that can further influence contaminant exposure. We compared contaminant bioaccumulation in four fish species, as well as two herbivorous invertebrates, from a coal combustion waste contaminated stream. Muscle, liver, and gonad tissue were analyzed from fish stratified across the broadest size ranges available. Effects of trophic position (δ (15)N), carbon sources (δ (13)C), and body size varied among and within species. Mercury and cesium concentrations were lowest in the invertebrates and increased with trophic level both among and within fish species. Other elements, such as vanadium, cadmium, barium, nickel, and lead, had greater levels in herbivorous invertebrates than in fish muscle. Sequestration by the fish livers averted accumulation in muscle. Consequently, fish liver tissue appeared to be a more sensitive indicator of bioavailability, but exceptions existed. Despite liver sequestration, within fishes, muscle concentrations of many elements still tended to increase by trophic level. Notable variation within some species was observed. These results illustrate the utility of stable isotope data in exploring differences of bioaccumulation within taxa. Our analyses suggest a need for further evaluation of the underlying sources of this variability to better understand contaminant bioaccumulation in large predatory fishes.

Effects of chronic copper exposure on development and survival in the southern leopard frog (Lithobates [Rana] sphenocephalus).

Exposure to environmental contaminants contributes to the global decline of amphibian populations. The impacts of organic contaminants on amphibians are well documented. However, substantially less is known concerning the potential effects of metals on amphibian populations. Copper (Cu) is an essential element, but it can be toxic at concentrations only slightly higher than the normal physiological range. The present study examines the effects of chronic Cu exposure on embryos and larvae of southern leopard frogs, Lithobates (Rana) sphenocephalus. Groups of eggs from multiple clutches were collected from two wetlands and exposed to a range of Cu concentrations (0-150 µg/L) until they reached the free-swimming stage, and then individual larvae were reared to metamorphosis. Higher Cu concentrations significantly reduced embryo survival to the free-swimming stage but did not further reduce survival to metamorphosis. Larval period was affected by Cu treatment, but the clutch from which larvae originated (i.e., parentage) explained a higher proportion of the variation. Embryo survival to hatching varied significantly among clutches, ranging from 42.9 to 79.2%. Measurable levels of Cu were found in larvae with body burdens up to 595 µg Cu/g dry mass in the 100 µg/L treatment, and larval Cu body burdens were higher than in metamorphs. The present study also demonstrated that higher initial egg density ameliorated embryo mortality at higher Cu levels and should be accounted for in future studies.

Comparative geochemical and microbiological characterization of two thermal pools in the Uzon Caldera, Kamchatka, Russia.

Arkashin Schurf (Arkashin) and Zavarzin Spring (Zavarzin), two active thermal pools in the Uzon Caldera, Kamchatka, Russia, were studied for geochemical and microbiological characterization. Arkashin, the smaller of the two pools, had broader temperature and pH ranges, and the sediments had higher concentrations of total As (4,250 mg/kg) relative to Zavarzin (48.9 mg/kg). Glycerol dialkyl glycerol tetraether profiles represented distinct archaeal communities in each pool and agreed well with previous studies of these pools. Although no archaeal 16S rRNA sequences were recovered from Arkashin, sequences recovered from Zavarzin were mostly representatives of the Crenarchaeota and "Korarchaeota," and 13% of the sequences were unclassifiable. The bacterial community in Arkashin was dominated by uncultured "Bacteroidetes," Hydrogenobaculum of the Aquificales and Variovorax of the Betaproteobacteria, and 19% of the sequences remained unclassified. These results were consistent with other studies of As-rich features. The most abundant members of the Zavarzin bacterial community included the Chloroflexi, as well as members of the classes Deltaproteobacteria and Clostridia. In addition, 24% of the sequences were unclassified and at least 5% of those represent new groups among the established Bacterial phyla. Ecological structure in each pool was inferred from taxonomic classifications and bulk stable isotope δ values of C, N, and S. Hydrogenobaculum was responsible for primary production in Arkashin. However, in Zavarzin, the carbon source appeared to be allochthonous to the identified bacterial community members. Additionally, sequences related to organisms expected to participate in N and S cycles were identified from both pools.

Phylogeny and historical biogeography of true morels (Morchella) reveals an early Cretaceous origin and high continental endemism and provincialism in the Holarctic.

True morels (Morchella, Ascomycota) are arguably the most highly-prized of the estimated 1.5 million fungi that inhabit our planet. Field guides treat these epicurean macrofungi as belonging to a few species with cosmopolitan distributions, but this hypothesis has not been tested. Prompted by the results of a growing number of molecular studies, which have shown many microbes exhibit strong biogeographic structure and cryptic speciation, we constructed a 4-gene dataset for 177 members of the Morchellaceae to elucidate their origin, evolutionary diversification and historical biogeography. Diversification time estimates place the origin of the Morchellaceae in the middle Triassic 243.63 (95% highest posterior density [HPD] interval: 169.35-319.89) million years ago (Mya) and the divergence of Morchella from its closest relatives in the early Cretaceous 129.61 (95% HPD interval: 90.26-173.16) Mya, both within western North America. Phylogenetic analyses identified three lineages within Morchella: a basal monotypic lineage represented by Morchella rufobrunnea, and two sister clades comprising the black morels (Elata Clade, 26 species) and yellow morels (Esculenta Clade, 16 species). Morchella possesses a Laurasian distribution with 37/41 species restricted to the Holarctic. All 33 Holarctic species represented by multiple collections exhibited continental endemism. Moreover, 16/18 North American and 13/15 Eurasian species appeared to exhibit provincialism. Although morel fruit bodies produce thousands of explosively discharged spores that are well suited to aerial dispersal, our results suggest that they are poorly adapted at invading novel niches. Morels also appear to have retained the ancestral fruit body plan, which has remained remarkably static since the Cretaceous.

Gracilibacter thermotolerans gen. nov., sp. nov., an anaerobic, thermotolerant bacterium from a constructed wetland receiving acid sulfate water.

An obligatorily anaerobic, thermotolerant, asporogenic bacterium, strain JW/YJL-S1(T), was isolated from a sediment sample of a constructed wetland system receiving acid sulfate water (pH 1.6-3.0). Cells of strain JW/YJL-S1(T) were straight to curved rods 0.2-0.4 mum in diameter and 2.0-7.0 mum in length, and stained Gram-negative. Growth of strain JW/YJL-S1(T) was observed at 25-54 degrees C (no growth at or below 20 or at or above 58 degrees C), with an optimum temperature range for growth of 42.5-46.5 degrees C. The pH(25 degrees C) range for growth was 6.0-8.25 (no growth at or below pH 5.7 or at or above pH 8.5), with optimum growth at pH 6.8-7.75. The salinity range for growth was 0-1.5 % (w/v) NaCl, with an optimum at 0-0.5 %. During growth on glucose the isolate produced acetate, lactate and ethanol as main fermentation end products. The fatty acid composition was dominated by branched-chain compounds: i15 : 0, a15 : 0, i16 : 0 and i17 : 0. The G+C content of the genomic DNA was 42.8 mol% (HPLC). Strain JW/YJL-S1(T) showed polymorphism of the 16S rRNA gene. Its closest relative was the thermophilic Clostridium thermosuccinogenes DSM 5807(T) (a member of Clostridium cluster III) (a blastn search revealed Clostridium pascui DSM 10365(T) to have 92.7 % gene sequence similarity, the highest value). The inferred phylogenetic trees placed strain JW/YJL-S1(T) between Clostridium clusters I/II and III. Based on the morphological and phylogenetic data presented, JW/YJL-S1(T) (=DSM 17427(T)=ATCC BAA-1219(T)) is proposed as the type strain of a novel species in a new genus, Gracilibacter thermotolerans gen. nov., sp. nov.

Thermosediminibacter oceani gen. nov., sp. nov. and Thermosediminibacter litoriperuensis sp. nov., new anaerobic thermophilic bacteria isolated from Peru Margin.

A new group of anaerobic thermophilic bacteria was isolated from enrichment cultures obtained from deep sea sediments of Peru Margin collected during Leg 201 of the Ocean Drilling Program. A total of ten isolates were obtained from cores of 1-2 m below seafloor (mbsf) incubated at 60 degrees C: three isolates came from the sediment 426 m below sea level with a surface temperature of 9 degrees C (Site 1227), one from 252 m below sea level with a temperature of 12 degrees C (Site 1228), and six isolates under sulfate-reducing condition from the lower slope of the Peru Trench (Site 1230). Strain JW/IW-1228P from the Site 1228 and strain JW/YJL-1230-7/2 from the Site 1230 were chosen as representatives of the two identified clades. Based on the 16S rDNA sequence analysis, these isolates represent a novel group with Thermovenabulum and Caldanaerobacter as their closest relatives. The temperature range for growth was 52-76 degrees C with an optimum at around 68 degrees C for JW/IW-1228P and 43-76 degrees C with an optimum at around 64 degrees C for JW/YJL-1230-7/2. The pH(25C) range for growth was from 6.3 to 9.3 with an optimum at 7.5 for JW/IW-1228P and from 5 to 9.5 with an optimum at 7.9-8.4 for JW/YJL-1230-7/2. The salinity range for growth was from 0% to 6% (w/v) for JW/IW-1228P and from 0% to 4.5% (w/v) for JW/YJL-1230-7/2. The G+C [corrected] mol% of the genomic DNA was 46.3 +/- 0.7% (n = 4) for Thermosediminibacter oceani [corrected] JW/IW-1228PT [corrected] and 45.2 +/- 0.7 (n = 6) for Thermosediminibacter litoriperuensis [corrected] JW/YJL-1230-7/2T [corrected] DNA-DNA hybridization yielded 52% similarity between the two strains. According to 16S rRNA gene sequence analysis, the isolates are located within the family, Thermoanaerobacteriaceae. Based on their morphological and physiological properties and phylogenetic analysis, it is proposed that strain JW/IW-1228P(T) is placed into a novel taxa, Thermosediminibacter oceani, gen. nov., sp. nov. (DSM 16646(T)=ATCC BAA-1034(T)), and JW/YJL-1230-7/2(T) into Thermosediminibacter litoriperuensis sp. nov. (DSM 16647(T) =ATCC BAA-1035(T)).

Lipid composition of suspended particulate matter (SPM) in a southeastern blackwater stream.

Suspended particulate matter (SPM) was collected seasonally for 1 yr at third- and fifth-order sites in a blackwater stream on the coastal plain in South Carolina. Fatty acids with carbon chain lengths from C(12)-C(32) were the most abundant component among the lipid classes examined with total concentrations varying from 8.5 to 60.2 microgL(-1). Unsaturated fatty acids predominated while significant concentrations of the even-chained saturated components C(24)-C(30) derived from cuticular plant waxes were also found. Concentrations of aliphatic alcohols, with carbon chain lengths between C(16) and C(30), ranged from 0.52 to 2.73 microgL(-1) and was dominated by the higher molecular weight compounds (C(22)-C(30)) derived primarily from cuticular plant waxes. Total hydrocarbon concentrations ranged from 0.35 to 5.66 microgL(-1) and showed no discernible trends with time or consistent difference between sites. The hydrocarbon assemblage observed indicates that these components are entirely of biogenic origin with no detectable anthropogenic contribution. The ratios of unsaturated to saturated fatty acids and cuticular to noncuticular fatty acids and alcohols at both stream locations indicate that the organic detritus associated with SPM collected during November and January is of more recent origin and less processed than that collected in the spring and summer months. Lipid concentrations at the third-order site generally had higher and more variable concentrations of the lipid classes compared with the fifth-order site. The ratio of saturated to unsaturated fatty acids was higher at the third-order site indicating that the organic fraction of SPM at this site was of more recent origin and less decomposed. The ratios of cuticular to noncuticular fatty acids and alcohols support this conclusion. These results indicate an export of particulate lipids of higher carbon resource quality from upstream to lower stream reaches.

Cyclooxygenase inhibitory and antioxidant compounds from the mycelia of the edible mushroom Grifola frondosa.

The bioassay-guided isolation and purification of the hexane extract of the cultured mycelia of Grifola frondosa led to the characterization of a fatty acid fraction and three compounds, ergosterol (1), ergostra-4,6,8(14),22-tetraen-3-one (2), and 1-oleoyl-2-linoleoyl-3-palmitoylglycerol (3). The composition of fatty acid fraction was confirmed as palmitic, oleic, and linoleic acids by GC-MS and by comparison with the retention values of authentic samples. The structures of compounds 1-3 were established by spectroscopic methods. The fatty acid fraction and compounds 1-3 showed cyclooxygenase (COX) enzyme inhibitory and antioxidant activities. The inhibition of COX-1 enzyme by the fatty acid fraction and compounds 1-3 at 250 microg/mL were 98, 37, 55, and 67%, respectively. Similarly, COX-2 enzyme activity was reduced by fatty acid fraction and compounds 1-3 at 250 microg/mL by 99, 37, 70, and 4%, respectively. The inhibitions of liposome peroxidation by the fatty acid fraction and compounds 1 and 2 at 100 microg/mL were 79, 48, and 42%, respectively. This is the first report of compounds 2 and 3 from the cultured mycelia of G. frondosa. The COX inhibitory activities of compounds 1-3 are reported here for the first time.