Halle, Universiẗat, Diss., 2000.

Halle, Wittenberg, Universiẗat, Diss., 2002.

Marie Malá, Isolation of alkaloids from Fontinalis antipyretica, thesis, Charles University in Prague, Faculty Of Pharmacy in Hradec Králové, 2012, 55 pages. The thesis Isolation of alkaloids from Fontinalis antipyretica, deals with quantification and isolation of substances of alkaloid character from above mentioned moss. The plant material collected in different localities in Czech Republic was extracted with methanol. The extract was subesquently used for isolation of alkaloids by HPLC method. The obtained amount of β-Carboline alkaloid was 2,4mg. The results of HPLC analysis (retention time, spectrum) of this sample were compared to harmalol standard and found the same. Finally the H NMR analysis proved, that the substance obtained by this experiment is not harmalol, but someother β-Carboline alkaloid.

Lucie Ptáčníková, The volume of alkaloids and flavonoids in Fontinalis antipyretica, Thesis, Charles University in Prague, Faculty of Pharmacy in Hradec Králové, thesis tutor: PharmDr. Jan Martin, PhD., Hradec Králové, 2012, 55 pages. Thesis called "Content of alkaloids and flavonoids in Fontinalis anitpyretica" establishes alkaloids and flavonoids and identifies one substance of these alkaloids in the above-mentioned water moss. The main purpose of this thesis was to obtain samples of Fontinalis antipyretica from selected locations in the Czech Republic (Ledce, Rychnovek, Kunvald). Next purpose was to determine alkaloids and flavonoids of collected material, isolated one substance of alkaloid character and verify identity of selected alkaloid. The content of alkaloids and flavonoids was determined by using the high-performance liquid chromatography (HPLC). The results show that β - carboline alkaloid harmin was determined by consensus UV spectrum and retention time with standard. Consequently flavonoids called myricetin and hyperosid was determined. Identity of β - carboline alkaloid harmin was verified by using NMR analysis. Keywords: alkaloids - harmalol - harmin - Fontinalis antipyretica - flavonoids

The brook trout <i>Salvelinus fontinalis</i> is the only salmonid native to the southern Appalachian Mountains, and is distributed across eastern North America from Canada to Georgia. This species was once abundant in coldwater lakes and streams throughout its range, but environmental disturbances and the introduction of non-native species have drastically reduced the number and sizes of wild populations. Genetic evidence suggests a division at the subspecies level between southern- and northern-derived brook trout populations, with the break between the two forms occurring roughly at the New River watershed. Before the subspecies structure was recognized, brook trout of northern origin were widely stocked throughout the Southeast. The primary objective of this study was to determine the phylogenetic origin of all wild brook trout populations in southwestern Virginia using allozyme markers. Seventy-eight streams believed to contain brook trout in the New, James, Holston, and Yadkin river drainages were sampled by backpack electrofishing. Muscle tissue samples were collected from 916 individuals from 56 populations using a non-lethal biopsy technique. The samples were analyzed by cellulose acetate gel electrophoresis and histochemical staining techniques. Variation at four polymorphic loci, including the diagnostic creatine kinase (<i>CK-A2*</i>) locus, was quantified in terms of genetic diversity and population genetic differentiation. Allele frequencies indicated that 19 populations were of putative southern origin, 5 of northern origin, and 32 of mixed genetic origin. The secondary objective was to determine the geographic distribution of southern- and northern-form brook trout populations throughout the native range using data compiled from all known genetic studies. A map of these data showed that the break between the southern and northern form is sharp, occurring at the New/Roanoke-James watershed divide. Populations from the New River drainage expressed the southern allele at a frequency of 85%, suggesting that their historic native character is southern, and that the presence of northern alleles is due to stocking or stream-capture events. The persistence of the southern form, despite the heavy stocking of northern-derived individuals, may be evidence of an adaptive advantage for the southern form of the species. Existence of adaptive genetic differentiation supports the case for conservation of the southern form of the species in future management of brook trout. / Master of Science

Habitat conditions, use and preference, size-at-age, density and condition were assessed for headwater brook charr (Salvelinus fontinalis) populations in streams in Southern Ontario where water extraction was occurring. Research was divided into two separate, but related studies, conducted during the summer and fall of 2002 and 2004. The first study sought to document the habitat conditions that influence size-at-age and density of age-0 brook charr and to test the hypotheses that: [1] differences in fork-lengths among sites were principally influenced by differences in stream thermal regimes as measured by accumulated thermal units (ATUs), with differences in stream physical characteristics (e.g., water quality, morphology, riparian vegetation) also having significant effects; [2] among-stream differences in flow regimes would significantly influence among-stream differences in age-0 densities, with increases in flow being positively correlated with density; and, [3] length and density would be negatively correlated. Results indicated that differences in temperature, stream site and flow influenced differences in age-0 length and density of brook charr. The effect of temperature, as measured by accumulated thermal units, dominated other influences, providing strong support for the null hypothesis [1] that size differences among sites were primarily driven by differences in the prevailing thermal regime. Some evidence was found to suggest that differences in stream physical characteristics influenced fish size and density, with stream flow affecting both to a moderate degree. Thus while study results provide support for the hypothesis [2] concerning flow-related impacts on age-0 brook charr in headwater streams, changes in flow cannot be considered of prime importance over the range of stream discharge differences (14.3-28.5 L???s -1) observed here. Finally, there was no support for hypothesis [3] concerning the negative correlation between age-0 fork-length and density. The second study investigated the effects of municipal groundwater pumping on stream habitat and the resident brook charr population of Monora Creek, Orangeville, Ontario. Specifically, the study aimed to test the hypotheses that: [1] well-related water extraction was associated with habitat degradation at sites proximate to the extraction point; and, [2] degraded habitat was associated with reduced juvenile and adult brook charr use and preference, reduced occupancy density and reduced condition. Study results indicated significant, although subtle, changes in available habitat as a result of pumping activity, with habitats in areas impacted by groundwater pumping showing significant decreases in hydraulic gradient, stream depth and the availability of under-cut bank and run and pool habitat. Combined, the changes in habitat influenced use, with age-0 densities declining in stream reaches impacted by pumping relative to reaches of the stream not impacted by pumping. When adjusting for the relative availability of groundwater extraction on affected and non-affected habitat, standardized habitat preference indices indicated a significant preference for non-affected stream reaches among age-0 brook charr. Adults evidenced only slight changes in distribution among the available habitats that were seasonally dependent, with reduced densities observed in affected habitats and increased densities in non-affected habitats noted only in the fall. Changes in brook charr condition (weight-length relationship) and mortality were generally not in evidence during the course of this study. Combined, these studies suggest development actions moderating either the thermal or flow regime of streams will have negative effects on population status. Water extraction risks altering habitat in unfavourable ways by interrupting important linkages between flow and temperature that favour growth and density, but that prevent critical physiological thresholds for temperature from being surpassed. Furthermore, the impacts on brook charr populations subjected to moderating pumping appear to relate more to patterns of within stream distribution and habitat use than to any direct measures of fitness such as survival or growth. The latter result suggests monitoring abundance and/or condition alone will not suffice to provide early warning of population stress responses to water extraction activities.

This diploma thesis is focused on determination of bioavailable cadmium in natural waters. Thesis is devided to two parts-laboratory study in model solution and experiments in real surface water system. During the laboratory experiments the accumulation and release constants of cadmium by aquatic moss Fontinalis antipyretica were determined. In Morava river experiment this aquatic moss was used to determine bioavailable fraction of cadmium and the moss bag technique were compared with another methods for phytotoxic metals determination.

Ce doctorat s’intègre dans le projet ANR BIOMET (JC05_59809) et a eu pour vocation d’améliorer les connaissances actuelles sur l’influence de la matière organique dissoute sur la spéciation et la biodisponibilité des métaux particulièrement dans le cas de système sous forte pression urbaine tel que la Seine. La compréhension de l’influence de la MOD sur la spéciation et la biodisponibilité des métaux suppose en outre que l’on connaisse précisément sa composition chimique ou plus exactement ses groupements fonctionnels. C’est pourquoi une partie de ces travaux est consacrée à l’extraction et la caractérisation de la matière organique dissoutes d’origine naturelle et urbaine. La caractérisation des fractions de MOD a été réalisée de manière originale par une approche multidimensionnelle à l’aide d’un véritable éventail de techniques analytiques (analyses élémentaires; isotopiques; fonctionnelles et moléculaires) permettant ainsi de mieux aborder l’influence de la MOD sur la spéciation et la biodisponibilité du cuivre et du plomb. Parallèlement à l’étude de caractérisation de la MOD, ce travail s’attache à mieux cerner le rôle de la MOD d’origine urbaine en particulier de la MOD hydrophile sur la spéciation et la biodisponibilité du cuivre et du plomb. Dans cet objectif, les techniques : potentiométrique à l’aide d’électrode ionique spécifique et la récente technique électrochimique AGNES ainsi que des biotests de toxicité aiguë (Daphnia magna) et de bioaccumulation (Fontinalis antipyretica) ont permis : - d’étudier la complexation du cuivre et du plomb par les fractions de MOD obtenues afin notamment de fournir des paramètres de complexations des MOD hydrophiles peu connues jusqu’à présent et ainsi mieux prédire le transport des métaux en milieu urbain. - d’évaluer le rôle protecteur de la MOD d’origine urbaine sur la biodisponibilité du cuivre. Les résultats ont montré que la MOD issue des effluents de station d’épuration et rejetée dans le milieu récepteur présente des caractéristiques originales à savoir un fort pourcentage de MOD hydrophile, c’est à dire une faible hydrophobicité et un degré d’aromaticité peu marqué soulignant le caractère peu réfractaire de cette MOD. En revanche une plus grande diversité fonctionnelle de la MOD d’origine urbaine par rapport à la MOD naturelle a été mise en évidence notamment par un taux de structures protéiques très important. Les résultats obtenus ici pour la première fois vis-à-vis des MOD hydrophiles d’origine urbaine ont montré une réactivité particulière vis-à-vis du cuivre. En effet les fractions HPI et plus généralement les fractions issues des effluents de station d’épuration ont montré une teneur en sites complexants plus élevée que ce qu’il avait été déjà observé pour des matières organiques naturelles. Les valeurs des constantes ne semblent pas varier d’une fraction à l’autre et assez peu suivant l’origine de la MOD. Le rôle particulier de l’azote dans la complexation du cuivre notamment sous la forme de groupements amines a été mis en évidence. Les approches utilisées pour évaluer la biodisponibilité du cuivre ont révélé pour l’ensemble des fractions de MOD un effet protecteur vis-à-vis du cuivre sur les organismes vivants. Néanmoins cet effet protecteur ne se trouve pas à la hauteur de celui pouvant être prédit par le modèle de l’ion libre principalement dû à une biodisponibilité de certains complexes organiques. Ceci peut être expliqué par l’important taux de structures protéiques dans ces MOD, les protéines étant connues pour jouer un rôle important dans les mécanismes de transports des métaux au sein des organismes. Par ailleurs, l’outil DGT (Diffusive gradient in thin films) a montré son efficacité pour évaluer la fraction biodisponible du cuivre / This thesis was carried out on the framework of the ANR BIOMET research project (JC05_59809). The main objectives were to improve the current knowledge on the influence of dissolved organic matter (DOM) on metal speciation and bioavailability in urban aquatic system. The understanding of the influence of DOM on trace metal speciation and bioavailability required that the chemical composition of DOM, more exactly its binding sites should be well understood. Therefore, one part of this research focused on the isolation and characterization of DOM. The characterization of DOM, with a multidimensional approach was realized with a lot of various techniques (such as elemental, isotopic, functional and molecular) that were of prime importance to better understand the influence of DOM on trace metal speciation and bioavailability. To accomplish these objectives, the techniques such as potentiometry with ionic selective electrode and the recent electroanalytical technique AGNES and bioassays such as a bioaccumulation test (Fontinalis antipyretica) and an acute toxicity test (Daphnia magna) were performed in order to: - study the copper and lead binding by isolated DOM fractions in order to give some binding parameters of hydrophilic DOM and to better predict the fate of the trace metal. - evaluate the protective role of urban DOM on copper bioavailability. Results showed some particular characteristics of DOM from wastewater effluent such as a high proportion of hydrophilic DOM, i.e. a low hydrophobicity and a low degree of aromaticity underlying the low refractory character of urban DOM. Nevertheless a higher content of various functional groups was determined in urban DOM than in natural DOM. A very high content of proteinaceous structures was particularly identified in urban DOM. Binding experiments revealed for the first time on urban hydrophilic DOM, a higher content of binding sites than in natural DOM and these sites are strongly correlated to amino groups. The values of binding constant seem to not vary according to the nature of the fraction or the origin. The different approaches used to evaluate copper bioavailability depicted a protective role of DOM to the organisms Daphnia magna and Fontinalis antipyretica. Nevertheless this protective effect is hardly explained by the free ion concentration due to a bioavailability of some organic complexes. This feature could be explain with the high content of proteinaceous structures in these DOM fractions, since proteins are well know to play an important role in the transport mechanisms of trace metals into the organisms. Moreover, the DGT device (diffusive gradient in thin films) shows a good efficiency to assess the bioavailable copper

Ce doctorat s'intègre dans le projet ANR BIOMET (JC05_59809) et a eu pour vocation d'améliorer les connaissances actuelles sur l'influence de la matière organique dissoute sur la spéciation et la biodisponibilité des métaux particulièrement dans le cas de système sous forte pression urbaine tel que la Seine. La compréhension de l'influence de la MOD sur la spéciation et la biodisponibilité des métaux suppose en outre que l'on connaisse précisément sa composition chimique ou plus exactement ses groupements fonctionnels. C'est pourquoi une partie de ces travaux est consacrée à l'extraction et la caractérisation de la matière organique dissoutes d'origine naturelle et urbaine. La caractérisation des fractions de MOD a été réalisée de manière originale par une approche multidimensionnelle à l'aide d'un véritable éventail de techniques analytiques (analyses élémentaires; isotopiques; fonctionnelles et moléculaires) permettant ainsi de mieux aborder l'influence de la MOD sur la spéciation et la biodisponibilité du cuivre et du plomb. Parallèlement à l'étude de caractérisation de la MOD, ce travail s'attache à mieux cerner le rôle de la MOD d'origine urbaine en particulier de la MOD hydrophile sur la spéciation et la biodisponibilité du cuivre et du plomb. Dans cet objectif, les techniques : potentiométrique à l'aide d'électrode ionique spécifique et la récente technique électrochimique AGNES ainsi que des biotests de toxicité aiguë (Daphnia magna) et de bioaccumulation (Fontinalis antipyretica) ont permis : - d'étudier la complexation du cuivre et du plomb par les fractions de MOD obtenues afin notamment de fournir des paramètres de complexations des MOD hydrophiles peu connues jusqu'à présent et ainsi mieux prédire le transport des métaux en milieu urbain. - d'évaluer le rôle protecteur de la MOD d'origine urbaine sur la biodisponibilité du cuivre. Les résultats ont montré que la MOD issue des effluents de station d'épuration et rejetée dans le milieu récepteur présente des caractéristiques originales à savoir un fort pourcentage de MOD hydrophile, c'est à dire une faible hydrophobicité et un degré d'aromaticité peu marqué soulignant le caractère peu réfractaire de cette MOD. En revanche une plus grande diversité fonctionnelle de la MOD d'origine urbaine par rapport à la MOD naturelle a été mise en évidence notamment par un taux de structures protéiques très important. Les résultats obtenus ici pour la première fois vis-à-vis des MOD hydrophiles d'origine urbaine ont montré une réactivité particulière vis-à-vis du cuivre. En effet les fractions HPI et plus généralement les fractions issues des effluents de station d'épuration ont montré une teneur en sites complexants plus élevée que ce qu'il avait été déjà observé pour des matières organiques naturelles. Les valeurs des constantes ne semblent pas varier d'une fraction à l'autre et assez peu suivant l'origine de la MOD. Le rôle particulier de l'azote dans la complexation du cuivre notamment sous la forme de groupements amines a été mis en évidence. Les approches utilisées pour évaluer la biodisponibilité du cuivre ont révélé pour l'ensemble des fractions de MOD un effet protecteur vis-à-vis du cuivre sur les organismes vivants. Néanmoins cet effet protecteur ne se trouve pas à la hauteur de celui pouvant être prédit par le modèle de l'ion libre principalement dû à une biodisponibilité de certains complexes organiques. Ceci peut être expliqué par l'important taux de structures protéiques dans ces MOD, les protéines étant connues pour jouer un rôle important dans les mécanismes de transports des métaux au sein des organismes. Par ailleurs, l'outil DGT (Diffusive gradient in thin films) a montré son efficacité pour évaluer la fraction biodisponible du cuivre

Wood ash application (WAA) has been recommended mainly for two reasons; i) to avoid depletion of minerals in the soil due to whole tree harvest in the forestry and ii) to mitigate harmful effects of acidification of soil and surface waters. In conclusion, the effects on terrestrial ecosystems and, especially, tree growth, can be attributed to the properties of the ash, the dose applied and the specific site at which the ash is applied. The research conducted on the effects of WAA on limnological ecosystems is very limited, and the major purpose of the present thesis was to gain knowledge of the effects of wood ash to different freshwater organisms, and the more comprehensive, limnological effects of WAA in the first stream in Bispgården, Sweden. Effects of wood ash solutions on the unicellular alga Euglena gracilis Klebs, the amphipod Gammarus pulex L., and the moss Fontinalis antipyretica Hedw. were investigated under laboratory conditions. Common in all three species was the decline in performance (growth/velocity/respiration/oxygen evolution) when the concentration of wood ash exceeded 5 g/l and no adjustment of pH was done (alkaline solution). In contrast, different movement parameters (motility, upwards swimming and velocity) in E. gracilis (neutral conditions), and increased growth of F. antipyretica with increased concentrations of wood ash indicated that nutrients in the ash was bioavailable for these organisms. There was no evidence of toxic effects on the organisms from metals or other compounds as a result from exposure to wood ash solutions in the present studies. The field study was conducted in a forest area close to Bispgården, about 100 km NW from Sundsvall, Sweden. The catchment area (50 ha) of the stream Fanbergsbäcken was treated with wood ash in September of 2004 (3,000 kg/ha;selfhardened crush-ash). In general, both biological (diatoms) and chemical (pH, alkalinity, and aluminum (Al) measurements) indicators have shown no significant effect on acidification parameters from the addition of wood ash. There was, however, evidence of an increased pH during spring flood, accompanied with a decrease in the frequency of low pH values (&lt;5.6) during spring flood. In addition to this, alkalinity was significantly higher in the period 2005-2006, compared to that of 2003. High concentrations of toxic forms of Al repeatedly occured in the stream Fanbergsbäcken, and the WAA did not affect the frequencies of high concentrations of toxic Al forms (&lt;50 μg/l). Both the moss F. antipyretica and the leaves from Alnus incana displayed increased potassium (K) concentrations, although other nutrients did not increase from WAA. In conclusion, no evidence of WAA being effective in restoring or improving the ecological status of freshwater environments has been established, either in the literature or in the present field study. On the other hand, there were no indications of harmful effects due to WAA, either. However, we still do not know the effects of wood ash on several organisms (predominantly invertebrates) inhabiting small ponds and other, temporary or permanent, freshwater ecosystems. In the context of WAA, these environments and organisms have not attended any attention in the research published to date, and future studies are strongly encouraged.