Screening and Identification of Coastal Chilean Thraustochytrids for Arachidonic Acid Production: Biotechnological Potential of Ulkenia visurgensis Lng2-Strain.

Thraustochytrids are unicellular heterotrophic marine protists that have been described as producing a high content of polyunsaturated fatty acids (PUFAs). Among them, arachidonic acid (ARA) stands out as a precursor of several mediators of pivotal importance for the immune system. However, the biotechnological potential of thraustochytrids for ARA production has not been developed. The objective of this study is to isolate and identify native strains from different Chilean coastal environments and evaluate in vitro the effect of culture parameters such as C/N ratio (19 and 33) and temperature (15 °C and 23 °C) on biomass production and arachidonic acid content. A total of nine strains were identified and classified into four genera of the Thraustochitridae family. The Lng2 strain with 99% identity belongs to the species Ulkenia visurgenis and was the most prominent one for ARA production. Temperature had an effect on the PUFA profile but not on the ARA content nor on the biomass yield. Additionally, the C/N ratio has been identified as a key parameter. The ARA productivity increased by 92% (from 0.6 to 8.3 ARA mg/g-DW) and its total biomass by 62.7% (from 1.9 to 5.1 g/L) at a high C/N ratio (33) as compared to the control.

Nitrogen fixation and other biogeochemically important features of Atacama Desert giant horsetail plant microbiomes inferred from metagenomic contig analysis.

BACKGROUND AND AIMS: Canyon stream beds in the hyperarid Atacama Desert surprisingly harbour magnificent groves of endemic giant horsetail wetland plants, Equisetum xylochaetum. Our previous metagenomic study of eukaryotes closely associated with this plant indicated that the microbiome included prokaryotes that might likewise influence host success and environment. We explored this possibility by using the metagenomic sequence to characterize prokaryote taxa and functional genes present in the microbiome of E. xylochaetum sampled from remote sites differing in the degree of anthropogenic disturbance. We focused on biogeochemical functions known to be important in wetland ecosystems. METHODS: To ensure that analyses were conducted on microbes most closely associated with plants, we extracted DNA from well-washed plant organs whose microbial biofilms were revealed with scanning electron microscopy. To assess the benefits of longer sequences for taxonomic and gene classifications, results of analyses performed using contigs were compared with those obtained with unassembled reads. We employed methods widely used to estimate genomic coverage of single taxa for genomic analysis to infer relative abundances of taxa and functional genes. KEY RESULTS: Key functional bacterial genera (e.g. Hydrogenophaga, Sulfuritalea and Rhodoferax) inferred from taxonomic and functional gene analysis of contigs - but not unassembled reads - to occur on surfaces of (or within) plants at relatively high abundance (>50× genomic coverage) indicated roles in nitrogen, sulfur and other mineral cycling processes. Comparison between sites revealed impacts on biogeochemical functions, e.g. reduced levels of the nifH gene marker under disturbance. Vanadium nitrogenases were more important than molybdenum nitrogenases, indicated by both functional genes and taxa such as Rhodomicrobium and Phaeospirillum inferred from contigs but not unassembled reads. CONCLUSIONS: Our contig-based metagenomic analyses revealed that microbes performing key wetland biogeochemical functions occur as tightly adherent biofilms on the plant body, not just in water or sediments, and that disturbance reduces such functions, providing arguments for conservation efforts.

Plastid Genome of Equisetum xylochaetum from the Atacama Desert, Chile and the Relationships of Equisetum Based on Frequently Used Plastid Genes and Network Analysis.

The modern pteridophyte genus Equisetum is the only survivor of Sphenopsida, an ancient clade known from the Devonian. This genus, of nearly worldwide distribution, comprises approximately 15 extant species. However, genomic information is limited. In this study, we assembled the complete chloroplast genome of the giant species Equisetum xylochaetum from a metagenomic sequence and compared the plastid genome structure and protein-coding regions with information available for two other Equisetum species using network analysis. Equisetum chloroplast genomes showed conserved traits of quadripartite structure, gene content, and gene order. Phylogenetic analysis based on plastome protein-coding regions corroborated previous reports that Equisetum is monophyletic, and that E. xylochaetum is more closely related to E. hyemale than to E. arvense. Single-gene phylogenetic estimation and haplotype analysis showed that E. xylochaetum belonged to the subgenus Hippochaete. Single-gene haplotype analysis revealed that E. arvense, E. hyemale, E. myriochaetum, and E. variegatum resolved more than one haplotype per species, suggesting the presence of a high diversity or a high mutation rate of the corresponding nucleotide sequence. Sequences from E. bogotense appeared as a distinct group of haplotypes representing the subgenus Paramochaete that diverged from Hippochaete and Equisetum. In addition, the taxa that were frequently located at the joint region of the map were E. scirpoides and E. pratense, suggesting the presence of some plastome characters among the Equiseum subgenera.

Removal of nutrients from Organic Liquid Agricultural Waste using filamentous algae / Remoção de nutrientes de Resíduos Agrícolas Líquidos Orgânicos utilizando algas filamentosas

Abstract A feasibility analysis of tertiary treatment for Organic Liquid Agricultural Waste is presented using filamentous algae belonging to the genus Cladophora sp. as an alternative to chemical tertiary treatment. The main advantages of tertiary treatments that use biological systems are the low cost investment and the minimal dependence on environmental variables. In this work we demonstrate that filamentous algae reduces the nutrient load of nitrate (circa 75%) and phosphate (circa 86%) from the organic waste effluents coming from dairy farms after nine days of culture, with the added advantage being that after the treatment period, algae removal can be achieved by simple procedures. Currently, the organic wastewater is discarded into fields and local streams. However, the algae can acquire value as a by-product since it has various uses as compost, cellulose, and biogas. A disadvantage of this system is that clean water must be used to achieve enough water transparency to allow algae growth. Even so, the nutrient reduction system of the organic effluents proposed is friendly to the ecosystem, compared to tertiary treatments that use chemicals to precipitate and collect nutrients such as nitrates and phosphates.

Resumo Uma análise de viabilidade do tratamento terciário para Resíduos Agrícola Líquidos Orgânicos é apresentada usando algas filamentosas pertencentes ao gênero Cladophora sp. como alternativa ao tratamento químico terciário. Os tratamentos terciários que utilizam sistemas biológicos têm baixo custo de investimento e a dependência de variáveis ​​ambientais é mínima. Neste trabalho, é demonstrado que essas algas filamentosas reduzem a carga nutricional de nitrato (circa 75%) e fosfato (circa 86%) dos efluentes de resíduos orgânicos provenientes de fazendas de leite em nove dias de cultura e tem a vantagem de que as algas podem ser facilmente coletadas posteriormente. Atualmente, as águas residuais orgânicas são descartadas nos campos e córregos locais. Posteriormente, as algas podem ser consideradas como matéria prima, uma vez que possuem várias utilidades como composto, celulose e biogás. Uma desvantagem desse sistema é que água limpa deve ser usada para obter transparência de água suficiente para permitir o crescimento de algas. Mesmo assim, o sistema de redução de nutrientes dos efluentes orgânicos propostos e amigável ao ecossistema, comparado aos tratamentos terciários que utilizam produtos químicos para precipitar e coletar nutrientes como nitratos e fosfatos.

Removal of nutrients from Organic Liquid Agricultural Waste using filamentous algae.

A feasibility analysis of tertiary treatment for Organic Liquid Agricultural Waste is presented using filamentous algae belonging to the genus Cladophora sp. as an alternative to chemical tertiary treatment. The main advantages of tertiary treatments that use biological systems are the low cost investment and the minimal dependence on environmental variables. In this work we demonstrate that filamentous algae reduces the nutrient load of nitrate (circa 75%) and phosphate (circa 86%) from the organic waste effluents coming from dairy farms after nine days of culture, with the added advantage being that after the treatment period, algae removal can be achieved by simple procedures. Currently, the organic wastewater is discarded into fields and local streams. However, the algae can acquire value as a by-product since it has various uses as compost, cellulose, and biogas. A disadvantage of this system is that clean water must be used to achieve enough water transparency to allow algae growth. Even so, the nutrient reduction system of the organic effluents proposed is friendly to the ecosystem, compared to tertiary treatments that use chemicals to precipitate and collect nutrients such as nitrates and phosphates.

Shotgun metagenomics and microscopy indicate diverse cyanophytes, other bacteria, and microeukaryotes in the epimicrobiota of a northern Chilean wetland Nostoc (Cyanobacteria).

Prokaryotic Nostoc, one of the world's most conspicuous and widespread algal genera (similar to eukaryotic algae, plants, and animals) is known to support a microbiome that influences host ecological roles. Past taxonomic characterizations of surface microbiota (epimicrobiota) of free-living Nostoc sampled from freshwater systems employed 16S rRNA genes, typically amplicons. We compared taxa identified from 16S, 18S, 23S, and 28S rRNA gene sequences filtered from shotgun metagenomic sequence and used microscopy to illuminate epimicrobiota diversity for Nostoc sampled from a wetland in the northern Chilean Altiplano. Phylogenetic analysis and rRNA gene sequence abundance estimates indicated that the host was related to Nostoc punctiforme PCC 73102. Epimicrobiota were inferred to include 18 epicyanobacterial genera or uncultured taxa, six epieukaryotic algal genera, and 66 anoxygenic bacterial genera, all having average genomic coverage ≥90X. The epicyanobacteria Geitlerinemia, Oscillatoria, Phormidium, and an uncultured taxon were detected only by 16S rRNA gene; Gloeobacter and Pseudanabaena were detected using 16S and 23S; and Phormididesmis, Neosynechococcus, Symphothece, Aphanizomenon, Nodularia, Spirulina, Nodosilinea, Synechococcus, Cyanobium, and Anabaena (the latter corroborated by microscopy), plus two uncultured cyanobacterial taxa (JSC12, O77) were detected only by 23S rRNA gene sequences. Three chlamydomonad and two heterotrophic stramenopiles genera were inferred from 18S; the streptophyte green alga Chaetosphaeridium globosum was detected by microscopy and 28S rRNA genes, but not 18S rRNA genes. Overall, >60% of epimicrobial taxa were detected by markers other than 16S rRNA genes. Some algal taxa observed microscopically were not detected from sequence data. Results indicate that multiple taxonomic markers derived from metagenomic sequence data and microscopy increase epimicrobiota detection.

Removal of nutrients from Organic Liquid Agricultural Waste using filamentous algae

Abstract A feasibility analysis of tertiary treatment for Organic Liquid Agricultural Waste is presented using filamentous algae belonging to the genus Cladophora sp. as an alternative to chemical tertiary treatment. The main advantages of tertiary treatments that use biological systems are the low cost investment and the minimal dependence on environmental variables. In this work we demonstrate that filamentous algae reduces the nutrient load of nitrate (circa 75%) and phosphate (circa 86%) from the organic waste effluents coming from dairy farms after nine days of culture, with the added advantage being that after the treatment period, algae removal can be achieved by simple procedures. Currently, the organic wastewater is discarded into fields and local streams. However, the algae can acquire value as a by-product since it has various uses as compost, cellulose, and biogas. A disadvantage of this system is that clean water must be used to achieve enough water transparency to allow algae growth. Even so, the nutrient reduction system of the organic effluents proposed is friendly to the ecosystem, compared to tertiary treatments that use chemicals to precipitate and collect nutrients such as nitrates and phosphates.

Resumo Uma análise de viabilidade do tratamento terciário para Resíduos Agrícola Líquidos Orgânicos é apresentada usando algas filamentosas pertencentes ao gênero Cladophora sp. como alternativa ao tratamento químico terciário. Os tratamentos terciários que utilizam sistemas biológicos têm baixo custo de investimento e a dependência de variáveis ambientais é mínima. Neste trabalho, é demonstrado que essas algas filamentosas reduzem a carga nutricional de nitrato (circa 75%) e fosfato (circa 86%) dos efluentes de resíduos orgânicos provenientes de fazendas de leite em nove dias de cultura e tem a vantagem de que as algas podem ser facilmente coletadas posteriormente. Atualmente, as águas residuais orgânicas são descartadas nos campos e córregos locais. Posteriormente, as algas podem ser consideradas como matéria prima, uma vez que possuem várias utilidades como composto, celulose e biogás. Uma desvantagem desse sistema é que água limpa deve ser usada para obter transparência de água suficiente para permitir o crescimento de algas. Mesmo assim, o sistema de redução de nutrientes dos efluentes orgânicos propostos e amigável ao ecossistema, comparado aos tratamentos terciários que utilizam produtos químicos para precipitar e coletar nutrientes como nitratos e fosfatos.

Impact of Cultivation Conditions, Ethylene Treatment, and Postharvest Storage on Selected Quality and Bioactivity Parameters of Kiwifruit "Hayward" Evaluated by Analytical and Chemometric Methods.

Organic, semiorganic, and conventional "Hayward" kiwifruits, treated with ethylene for 24 h and stored during 10 days, were assessed by UV spectrometry, fluorometry, and chemometrical analysis for changes in selected characteristics of quality (firmness, dry matter and soluble solid contents, pH, and acidity) and bioactivity (concentration of polyphenols via Folin-Ciocalteu and p-hydroxybenzoic acid assays). All of the monitored qualitative parameters and characteristics related to bioactivity were affected either by cultivation practices or by ethylene treatment and storage. Results obtained, supported by statistical evaluation (Friedman two-way ANOVA) and chemometric analysis, clearly proved that the most significant impact on the majority of the evaluated parameters of quality and bioactivity of "Hayward" kiwifruit had the ethylene treatment followed by the cultivation practices and the postharvest storage. Total concentration of polyphenols expressed via p-hydroxybenzoic acid assay exhibited the most significant sensitivity to all three evaluated parameters, reaching a 16.5% increase for fresh organic compared to a conventional control sample. As a result of postharvest storage coupled with ethylene treatment, the difference increased to 26.3%. Three-dimensional fluorescence showed differences in the position of the main peaks and their fluorescence intensity for conventional, semiorganic, and organic kiwifruits in comparison with ethylene nontreated samples.

Comprehensive two-dimensional gas chromatography and three-dimensional fluorometry for detection of volatile and bioactive substances in some berries.

The volatile fractions of Cape gooseberry and blueberry were determined by headspace solid-phase microextraction coupled with comprehensive two-dimensional gas chromatography with time-of-flight mass spectrometry (HS-SPME/GC×GC-TOFMS). The highest amount of alcohol (51.8%), ester (32.8%) and carboxylic acid (6.9%) was in blueberry in comparison with gooseberry and oppositely ketones (14.7%), aldehydes (9.9%) and terpenes (8%) were found in gooseberry. The bioactive compounds and antioxidant capacities were higher in blueberries than in gooseberries. Three dimensional fluorescence emission spectrometry (3D-FL) was applied to determine and to compare experimentally found binding parameters of berries extracts with human serum albumin (HSA). The fluorescence quenching of HSA by polyphenols from berries was a result of the formation of a polyphenol-HSA complex. The binding abilities of berries were highly correlated with the bioactivity of polyphenols and volatile substances. The cluster analysis (CA) and linear discriminant analysis (LDA) was applied to differentiate the berries samples according to their type.

Lacustrine Nostoc (Nostocales) and associated microbiome generate a new type of modern clotted microbialite.

Microbialites are mineral formations formed by microbial communities that are often dominated by cyanobacteria. Carbonate microbialites, known from Proterozoic times through the present, are recognized for sequestering globally significant amounts of inorganic carbon. Recent ecological work has focused on microbial communities dominated by cyanobacteria that produce microbial mats and laminate microbialites (stromatolites). However, the taxonomic composition and functions of microbial communities that generate distinctive clotted microbialites (thrombolites) are less well understood. Here, microscopy and deep shotgun sequencing were used to characterize the microbiome (microbial taxa and their genomes) associated with a single cyanobacterial host linked by 16S sequences to Nostoc commune Vaucher ex Bornet & Flahault, which dominates abundant littoral clotted microbialites in shallow, subpolar, freshwater Laguna Larga in southern Chile. Microscopy and energy-dispersive X-ray spectroscopy suggested the hypothesis that adherent hollow carbonate spheres typical of the clotted microbialite begin development on the rigid curved outer surfaces of the Nostoc balls. A surface biofilm included >50 nonoxygenic bacterial genera (taxa other than Nostoc) that indicate diverse ecological functions. The Laguna Larga Nostoc microbiome included the sulfate reducers Desulfomicrobium and Sulfospirillum and genes encoding all known proteins specific to sulfate reduction, a process known to facilitate carbonate deposition by increasing pH. Sequences indicating presence of nostocalean and other types of nifH, nostocalean sulfide:ferredoxin oxidoreductase (indicating anoxygenic photosynthesis), and biosynthetic pathways for the secondary products scytonemin, mycosporine, and microviridin toxin were identified. These results allow comparisons with microbiota and microbiomes of other algae and illuminate biogeochemical roles of ancient microbialites.

Characterization of Rapana thomasiana as an indicator of environmental quality of the Black Sea coast of Bulgaria.

The aim of this investigation was to determine the contents of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), minerals, trace elements and bioactivity in the gastropod Rapana thomasiana, which can be used as an environmental bioindicator organism. The chemical differences between Rapana thomasiana from polluted (RapaPol) and non-polluted (RapaNPol) sites of the Black Sea coast in Bulgarian were investigated. Chromatography and high-resolution inductively coupled plasma mass spectrometry (HR-ICP-MS) were used for evaluation of PAHs, PCBs, minerals and trace elements. Methanol extracts from RapaPol and RapaNPol (to a lesser degree) contained relatively high amounts of free phenolics (2.50 +/- 0.3 and 1.57 +/- 0.18 mg GAE/g DW, respectively) and exhibited the following respective levels of antioxidant activities determined by two radical-scavenging assays (microMTE/g DW): 1.8 +/- 0.2 and 0.98 +/- 0.08 by 1-diphenyl-2-picrylhydrazyl method (DPPH); 1.74 +/- 0.17 and 1.04 +/- 0.12 by cupric reducing antioxidant capacity (CUPRAC). The total amounts of elements, PAHs and PCBs were higher in RapaPol than in RapaNPol. The obtained indices of Rapana thomasiana can serve as a bioindicator of the environmental ecological quality.

Evaluation of inhibition of cancer cell proliferation in vitro with different berries and correlation with their antioxidant levels by advanced analytical methods.

Dimethylsulfoxide extracts of Chilean berries [Myrteola nummularia, 'Murtilla-like'] vs. well known 'Murtilla', Chilean and Polish blueberries, and Chilean raspberries were investigated for their antioxidant, quenching and antiproliferative activities. The significantly highest levels of polyphenols, flavonoids, flavanols and their antioxidant activities were estimated in 'Murtilla' (MT) berries (P<0.05), than in other investigated samples. DPPH kinetic measurements were calculated to compare, distinguish and discriminate the antiradical activity among berry extracts by multivariate analysis. The lowest IC(50) values, 751 and 858 µg/ml, were obtained for MT extract on colon cancer cell lines HT-29 and SW48. HT-29 cells treated with MT extract showed a decrease in G1 phase cells from 77% to 56% (P<0.05). At the highest concentration of 2000 µg/ml MT extract caused 90-100% cell growth inhibition. Percentage of death cells treated with MT extract was 80.1% and 72.5% for SW48 and HT-29 cells, respectively. The inhibition of cancer cell proliferation highly correlated with the levels of polyphenols, flavonoids and their antioxidant activities. The interaction between drugs and serum albumin plays an important role in the distribution and metabolism of drugs, therefore the complexation reaction between flavonoids, and berries extracts, and bovine serum albumin (BSA) was investigated by 3-D fluorescence and FTIR spectroscopy. The results indicated that flavonoids and polyphenol extracts have strong ability to quench the intrinsic fluorescence of BSA by forming complexes. A shift in the maximum of amides FTIR-bands appeared. In conclusion, these findings suggest that the intake of a new kind of berry, as a source of natural antioxidants, may reduce colon cancer risk.

Aeroterrestrial Coleochaete (Streptophyta, Coleochaetales) models early plant adaptation to land.

PREMISE OF THE STUDY: The streptophyte water-to-land transition was a pivotal, but poorly understood event in Earth history. While some early-diverging modern streptophyte algae are aeroterrestrial (living in subaerial habitats), aeroterrestrial survival had not been tested for Coleochaete, widely regarded as obligately aquatic and one of the extant green algal genera most closely related to embryophytes. This relationship motivated a comparison of aeroterrestrial Coleochaete to lower Paleozoic microfossils whose relationships have been uncertain. METHODS: We tested the ability of two species of the experimentally tractable, complex streptophyte algal genus Coleochaete Bréb. to (1) grow and reproduce when cultivated under conditions that mimic humid subaerial habitats, (2) survive desiccation for some period of time, and (3) produce degradation-resistant remains comparable to enigmatic Cambrian microfossils. KEY RESULTS: When grown on mineral agar media or on quartz sand, both species displayed bodies structurally distinct from those expressed in aquatic habitats. Aeroterrestrial Coleochaete occurred as hairless, multistratose, hemispherical bodies having unistratose lobes or irregular clusters of cells with thick, layered, and chemically resistant walls that resemble certain enigmatic lower Paleozoic microfossils. Whether grown under humid conditions or air-dried for a week, then exposed to liquid water, aeroterrestrial Coleochaete produced typical asexual zoospores and germlings. Cells that had been air-dried for periods up to several months maintained their integrity and green pigmentation. CONCLUSIONS: Features of modern aeroterrestrial Coleochaete suggest that ancient complex streptophyte algae could grow and reproduce in moist subaerial habitats, persist through periods of desiccation, and leave behind distinctive microfossil remains.

Determination of PAHs, PCBs, minerals, trace elements, and fatty acids in Rapana thomasiana as an indication of pollution.

The aim of this investigation was to introduce several analytical methods for determination of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), minerals, trace elements, and fatty acids in Rapana thomasiana as a marine pollution indicator organism. The chemical differences of the gastropod Ra. thomasiana from polluted and nonpolluted sites of the Black Sea on the Bulgarian coast were investigated. Chromatography and high-resolution inductively coupled plasma/MS analyses were used for evaluation of PAHs, PCBs, fatty acids, minerals, and trace elements. These methods can be applied to other marine products.

Evolutionary and ecophysiological significance of sugar utilization by the peat moss Sphagnum compactum (Sphagnaceae) and the common charophycean associates Cylindrocystis brebissonii and Mougeotia sp. (Zygnemataceae).

PREMISE OF THE STUDY: The goal of this study was to illuminate the evolutionary history and ecological importance of plant mixotrophy-the uptake and utilization of exogenous organic compounds. • METHODS: We quantitatively assessed the effect of sugar amendments on laboratory growth of Sphagnum compactum as a representative emergent peat moss and two species of ecologically associated zygnematalean algae, Cylindrocystis brebissonii and Mougeotia sp. • KEY RESULTS: Together with observations published elsewhere, our results suggest that under carbon or light limitation, the uptake of exogenous sugars by cells of charophycean algae and peat mosses may help these organisms maintain positive carbon balance. Utilization of 1% glucose by aquatic-grown algae helped to relieve dissolved inorganic carbon limitation, enhancing photoautotrophic growth by factors of 9.0 and 1.7, respectively. After an 8-wk growth period, amendments of 1% and 2% glucose enhanced air-grown moss biomass by 28 and 39 times, respectively, that of controls lacking sugar amendments. After 9 wk, 1% fructose enhanced biomass by 21 times, and 2% sucrose enhanced biomass by 31 times. • CONCLUSION: Our results indicate that plant mixotrophy is an early-evolved trait. The results also indicate that quantitative differences in sugar utilization by bryophytes and charophycean algae correlate with relative investments in protective cell-wall polyphenolics measured in previous studies, suggesting that sugar utilization may subsidize the cost of producing phenolic wall compounds in bryophytes.

Screening of the antioxidant and nutritional properties, phenolic contents and proteins of five durian cultivars.

The antioxidant activities of different durian cultivars at the same stage of ripening (Mon Thong, Chani, Kan Yao, Pung Manee and Kradum) were compared in order to choose the best as a supplement in the human diet. Total polyphenols (mg gallic acid equivalent/100 g fresh weight (FW)) and flavonoids (mg catechin equivalent (CE)/100 g FW) in Mon Thong (361.4+/-23.2 and 93.9+/-7.4) were significantly higher (P<0.05) than in Kradum (271.5+/-11.2 and 69.2+/-5.3) and Kan Yao (283.2+/-16.5 and 72.1+/-6.8). The free polyphenols and flavonoids showed lower results than the hydrolyzed ones. Anthocyanins (microg cyanidin-3-glucoside equivalent/100 g FW) and flavanols (microg CE/100 g FW) were significantly higher in Mon Thong (427.3+/-23.8 and 171.4+/-16.3) than in Kradum (320.2+/-12.1 and 128.6+/-9.7) and Kan Yao (335.3+/-14.1 and 134.4+/-11.7). Ultraviolet spectroscopy and high-performance liquid chromatography/diode array detection analyses showed that caffeic acid and quercetin were the dominant bioactive substances in Mon Thong cultivar. The antioxidant activity (microM trolox equivalent/100 g FW) of Mon Thong cultivar (260.8+/-20.2, 1,075.6+/-81.4 and 2,352.7+/-124.2) determined by ferric-reducing/antioxidant power (FRAP), cupric reducing antioxidant capacity (CUPRAC) and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) with Trolox equivalent antioxidant capacity (TEAC) assays was significantly higher (P<0.05) than in Kradum (197.4+/-8.9, 806.5+/-31.2 and 1,773.2+/-102.5) and in Kan Yao (204.7+/-9.7, 845.5+/-48.6 and 1,843.6+/-107.5). The correlation coefficients between polyphenols, flavonoids, flavanols and FRAP, CUPRAC and TEAC capacities were between 0.89 and 0.98. In extracted and separated by electrophoresis durian proteins, some differences were found in the sodium dodecyl sulfate-protein bands in the region of 16 and 68 kDa for Kradum, 45 kDa for Mon Thong and three bands for Kan Yao. Antioxidants and proteins can be used for characterization of the quality of durian cultivars. In conclusion, the bioactivity of durian cultivars Mon Thong, Chani and Pung Manee was high and the total polyphenols were the main contributors to the overall antioxidant capacity. The results of our investigation in vitro are comparable with other fruits that widely used in human diets. Therefore, Durian can be used as a supplement for nutritional and healthy purposes, especially Durian Mon Thong, Chani and Pung Manee.

The bioactivity of processed garlic (Allium sativum L.) as shown in vitro and in vivo studies on rats.

Garlic (Allium sativum L.) is widely used as an obligatory part in many cooked dishes loosing during this process a certain part of its bioactivity. Antioxidant capacity measured by the ferric-reducing/antioxidant power (FRAP) method and by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical assay was the highest in raw and in a short time processed garlic samples by cooking. 70 Wistar rats were randomly divided into 10 diet groups, each of seven. They were named Control, NPG, PG1, PG2, PG3, Chol, Chol/NPG, Chol/PG1, Chol/PG2 and Chol/PG3. The rats of the Control group were fed basal diet (BD), which included wheat starch, casein, soybean oil, cellulose, mineral and vitamin mixtures. To the BD of the nine other groups were added 25mg of lyophilized non processed garlic equivalent of 500 mg non processed garlic/kg body weight (NPG), the same quantity of processed garlic for 20, 40 and 60 min for PG1, PG2 and PG3, respectively, 1% of cholesterol (Chol), 1% of cholesterol and 25mg/kg body weight of lyophilized non processed garlic (Chol/NPG), 1% of cholesterol and the same quantity of processed garlic for 20, 40 and 60 min for Chol/PG1, Chol/PG2 and Chol/PG3, respectively. The dose of 500 mg (25mg of lyophilized garlic/kg body weight) was chosen as the most effective (Banerjee, S.K., Maulik, M., Mancahanda, S.C., Dinda, A.K., Gupta, S.K., Maulik, S.K., 2002. Dose-dependent induction of endogenous antioxidants in rat heart by chronic administration of garlic. Life Sciences 70, pp. 1509-1518). Plasma lipid profile and the total antioxidant capacity in rats significantly differed in diet groups with addition of garlic samples cooked for a long time. In summary, garlic cooked for a short time preserves a high bioactivity of non processed garlic. The diet supplemented with these samples and cholesterol improved lipid indices, decreased fibrinogen and increased antioxidant activity in plasma of rats. Therefore, for preservation of garlic bioactivity optimal regime has to be used.

Relationship between seawater pollution and qualitative changes in the extracted proteins from mussels Mytilus galloprovincialis.

The aim of this study was to find a reliable biomarker of seawater pollution. For this purpose the contents of Zn and Cu, proteins and antioxidant activity in mussels Mytilus galloprovincialis collected from polluted and non-polluted sites of the Bulgarian Black Sea coast were compared. To determine the above-mentioned indices atomic spectroscopy, Fourier Transform Infrared (FT-IR) spectroscopy, fluorescence, differential scanning calorimetry (DSC), and two antioxidant tests were used. It was found that the amounts of Zn and Cu were significantly higher in the mussel proteins from the polluted than from the non-polluted sites (P<0.05). FT-IR spectroscopy and fluorescence revealed specific qualitative changes in secondary and tertiary structures in mussel proteins in the samples from polluted sites. The thermodynamic properties of proteins and the changes upon denaturation were correlated with the secondary structure of proteins and disappearance of alpha-helix. Purified protein scavenging activity against 2, 2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) radical cation (ABTS(*+)) was significantly higher in mussel samples from polluted than from non-polluted sites. Therefore, the changes in Zn and Cu concentration, in protein's secondary and tertiary structures and antioxidant activity in mussels M. galloprovincialis from polluted sites can be a reliable biomarker of the level of the seawater pollution.

Changes in mussel Mytilus galloprovincialis protein profile as a reaction of water pollution.

In the present report the changes in mussels Mytilus galloprovincialis protein profile, as a reaction of water pollution, is presented. The antioxidant capacity was determined by the efficacy to scavenge the derived radical in mussel samples using the Trolox Equivalent Antioxidant Capacity (TEAC) assay. The highest percentage of inhibition was estimated in protein mussels from the dirty area (DM) in comparison with the clean samples (CM). The amounts of both zinc and copper determined by atomic spectroscopy were significantly higher in the protein mussels from the dirty than from the clean areas (P<0.05). Fourier Transform Infrared (FT-IR) spectroscopy and fluorescence revealed specific qualitative changes in secondary and tertiary structures of mussel proteins from the dirty area by the shift in the amides I and II positions and fluorescence intensity. Estimated temperature and enthalpy of denaturation in the protein mussels well correlated with fluorescence and spectroscopic measurements and showed the changes influenced by water pollution.

Production and purification of statins from Pleurotus ostreatus (Basidiomycetes) strains.

Pleurotus ostreatus strains were cultured in liquid medium and on wheat straw. The yields of lovastatin were compared.