This diploma thesis deals with design and experimental study of transdermal transport of pharmaceutically active agents from gel matrices, which contain humic substance in its structure. A model absorption membrane was represented by the skin of pig´s earlobes. The study of the release of active substances and Lignohumte was performed due to the vertical diffusion cells. The amount of released humic substance was characterized by UV-VIS method and the amount of released active agent from gel matrix was determined by HPLC-DAD. The most interesting finding of this diploma thesis was that Lignohumate enhances transdermal transport of active agents and supports their release from gel samples to the particular environment.

F�r die Therapie und Prophylaxe von Fischkrankheiten stehen nur wenige geeignete und zugelassene Medikamente zur Verf�gung. Daher kommt der Suche nach wirksamen und f�r Tier, Verbraucher und Umwelt sicheren Alternativen in der Fischhaltung eine besondere Bedeutung zu. �ber positive Effekte beim oralen Einsatz von Huminstoffen an landwirtschaftlichen Nutztieren, Zootieren und Heimtieren liegen eine Vielzahl von Berichten vor. Deshalb sollten in der vorliegenden Studie Huminstoffe (HS) in der �u�erlichen Anwendung bei Zierfischen und in der oralen Verabreichung �ber das Futter bei Nutzfischen n�her untersucht werden. Dazu wurden zwei Versuche (B I: Dauer 4 Tage; B II: Dauer 12 Tage) an Goldfischen (Carassius auratus) unter teichwirtschaftlichen Bedingungen, zwei Aufzuchtversuche (F I: Kv bis K3, Dauer: 1.000 Tage: F II: K0 bis K1, Dauer: 150 Tage) an Karpfen (Cyprinus carpio) in der Teichwirtschaft und ein Aufzuchtversuch (F III: vorgestreckte Brut bis 30 g Lebendmasse, 166 Tage) an Regenbogenforellen (Oncorhynchus mykiss) unter Laborbedingungen durchgef�hrt. Als Versuchsparameter dienten Mortalit�t und Morbidit�t, Wachstum und Futteraufwand in den Aufzuchtversuchen, pathologisch anatomische (einschlie�lich histologischer Befunde in B I und F II), parasitologische, bakteriologische und virologische Kriterien. Die Wasserqualit�t wurde regelm��ig ermittelt. In den Versuchen B I und B II wurde eine 10%ige Na-Humat-L�sung (HS vom Typ WH 67 SNa der Fa. Pharmawerk Weinb�hla) dem H�lterungswasser so zugesetzt, dass eine Na-Humat-Konzentration von 10 mg L-1 entstand. Zweij�hrige, multifaktoriell erkrankte Goldfische (B I: N=2 x 322, B II: N=2 x 100) wurden dem Na-Humat-Bad t�glich f�r die Dauer einer Stunde (B I) bzw. von 1,5 bis 2 Stunden (B II) ausgesetzt. Die HS-Badehandlungen f�hrten in beiden Versuchen zu einer Senkung der Verluste (Mortalit�tsrate in B I: 0,6 versus 4,7 %, p<0,0005; B II: 8 versus 16 %, p<0,05). Bei Versuchsende war in den Versuchsgruppen (Na-Humat-Bad) der Gesundheitszustand, beurteilt nach �u�erlich sichtbaren Ver�nderungen an Haut, Flossen und Kiemen, deutlich verbessert (B I: p<1�10-16; B II: p<0,001). So waren (Signifikanzangaben nur f�r B II) vor allem H�morrhagien der Haut und der Flossen (p<0,05), Flossenrand- und Kiemennekrosen (p<0,05) sowie akute Kiemenentz�ndungen (p<0,02) verringert. Die Badebehandlungen bewirkten weiter eine geringere Auspr�gung integument�rer Mykosen der Haut, Flossen und Kiemen durch Pilze der Gattung Saprolegnia (p<0,05). Der Befall von Haut und Kiemen mit vorrangig protozo�ren Ektoparasiten konnte insgesamt ebenso vermindert werden (p<0,01) wie der Befall der Kiemen mit Ichthyophthirius sp. (p<0,05). Aus der beobachteten Milderung von Krankheitssymptomen umweltbedingter, bakterieller, ektoparasit�rer und mykotischer Genese nach Na-Humat-B�dern bei Goldfischen kann auf einen gewebeprotektiven, antiphlogistischen, antimykotischen und antiparasit�ren Effekt der Na-Humat-Behandlung geschlossen werden. In der Aufzucht von Karpfen (F I) und Regenbogenforellen (F III) wurden dem Futter 5 % Humocarb, formuliert� (HS vom Typ WH 67 A), im Karpfenversuch F II 5 % Cellu-Ligno-Karbon-Isolat (HS vom Typ WH 67 G, beide Produkte: Fa. Pharmawerk Weinb�hla) zugesetzt. In F I und F III wurde ein gleiches pelletiertes Alleinfutter f�r Forellen, in F II ein pelletiertes Alleinfutter f�r Karpfen eingesetzt. In Versuch F I (Besatzdichte der beiden Teiche (0,2 ha) jeweils 35.000 Kv ha-1) wiesen die Karpfen der Versuchs- gegen�ber der Kontrollgruppe h�here mittlere K�rpermassen auf. Der Unterschied zwischen den Gruppen bewegte sich mit einer Ausnahme (zu geringer Stichprobenumfang) zwischen p<0,05 (65 versus 45 g, 114. Tag) und p<0,0001 (1.146 versus 855 g, Versuchsende 1000. Tag). Der um die HS-Aufnahme bereinigte Futteraufwand des ersten Versuchsjahres war durch HS um 34 % verbessert. Die Futterquotienten in der zweiten und dritten Aufzuchtperiode waren durch die subjektive Futterzumessung (Nichtber�cksichtigung zwischenzeitlicher Verluste) verf�lscht. In Versuch F II (Besatzdichten der 2 x 3 Teiche (0,25 ha) 20.000, 30.000 bzw. 40.000 K0 ha-1) erhielten die Br�tlinge �ber die ersten zwei Wochen kein zus�tzliches Futter (nur Naturnahrung), anschlie�end wurde Weizenschrot zugef�ttert, erst ab dem 63. Tag das Versuchs- bzw. Kontrollfutter f�r 87 Tage. Bei Versuchsende war in den Gruppen der h�chsten Besatzdichte die Mortalit�t durch HS um ca. 20 % verringert (p<0,0001). Die Abfischungsmassen waren bei geringer Besatzdichte ann�hernd gleich, bei mittlerer Besatzdichte war die Kontrollgruppe, bei h�chster Besatzdichte die Versuchsgruppe favorisiert (statistische Pr�fung wegen des Fehlens von Einzeldaten nicht m�glich). Der um die HS-Aufnahme bereinigte Futteraufwand war in den HS-Gruppen bei geringer bzw. h�chster Besatzdichte um 5 bzw. 17 % verringert. In beiden Versuchen, F I und F II, wurde eine deutliche Verringerung des Gehaltes der inneren Organe Leber, Niere und Milz an fakultativ pathogenen Aeromonas- und Pseudomonas spp. nachgewiesen (p<0,05). Bei der Aufzucht von Regenbogenforellen (F III; N=2 x 25) trugen die durch den HS-Zusatz bedingten Ver�nderungen der Konsistenz von Futter und Kot zur Verminderung einer starken Belagbildung (Bewuchs aus Algen und anderen Mikroorganismen) sowie zur Vermeidung von Wassertr�bungen in den Aquarien, insbesondere bei h�heren Temperaturen, bei. Signifikante Unterschiede in den Kenndaten von Wachstum und Futteraufwand wurden nicht beobachtet. Insgesamt gesehen erscheint aber der Zusatz von 5 % HS zum Futter als zu hoch gew�hlt, da sich der Futteraufwand in den HS-Gruppen bei Ber�cksichtigung der Gesamtfuttermenge (einschlie�lich des 5%igen HS-Zusatzes) �berwiegend ung�nstiger gestaltete. Die Effekte der oralen Applikation von 5 % HS im Futter sind damit weit weniger augenf�llig als nach �u�erlicher Anwendung. Die zootechnischen Parameter waren �berwiegend mehr oder weniger unver�ndert; allein in Versuch F I war die K�rpermasse in der HS-Gruppen im Versuchsverlauf (Stichproben und Abfischung) in der Regel signifikant h�her als in der Kontrollgruppe. Von k�nftiger Bedeutung f�r die Aquarienhaltung von Fischen (und m�glicherweise in Klein- und Zierteichen) k�nnte die beobachtete Wasserkl�rung und die Verminderung einer Belagbildung aus Algen und anderen Mikroorganismen sein. Eine intestinale Wirkung der HS wird insoweit unterstellt, als in beiden Karpfenversuchen die inneren Organe einen verminderten Befall an beweglichen Aeromonaden und Pseudomonaden aufwiesen. / Prophylactic and therapeutic use of brown coal-derived humic substances of the type WH 67 in finfish and ornamental fish farming On the market, there is a limited number of suitable and approved drugs available for therapy and prophylaxis of fish diseases. The search for alternatives which are both effective and safe for animals, consumers and environment is therefore of special importance. There are numerous reports on positive effects of orally administered humic substances (HS) on agricultural livestock, zoo animals and pets. Therefore, the present study aimed on effects of HS for both external treatment of ornamental fish and oral application on culture fish via feed. Two experiments on goldfish (Carassius auratus) (exp. B I: 4 days; exp. B II: 12 days), and on common carp (Cyprinus carpio) (exp. F I: 1,000 days, fry to 3-year-old fish ; exp. F II: 150 days, fry to young-of-the-year size) were carried out under pond culture conditions. Additionally, one experiment on rainbow trout (Oncorhynchus mykiss) was carried out in the laboratory (exp. F III: 166 days, fingerlings up to 30 g body weight). The following parameters were examined: mortality, morbidity, growth, feed conversion ratio (exp. F I, F II, F III only), pathologic anatomical criteria (including histological examination of fish from B I and F II), parasitological investigation, bacteriological and virological state. Water quality was regularly examined as well. In the experiments B I and B II, a 10% sodium-humate solution (type WH 67 SNa, Pharmawerk Weinb�hla) was mixed into the water until a sodium-humate concentration of 10 mg L-1 was reached. Two-year old, multifactorially diseased goldfish (B I: N= 2 x 322; B II: N= 2 x 100) were daily exposed to this water for a period of one (B I) and 1.5-2 (B II) hours, respectively. In both experiments the treatment of fish reduced in mortality (B I: 0.6 vs. 4.7 %, p<0.0005; B II: 8 vs. 16 %, p<0.05). Moreover, at the end of the experiments the sodium-humate treated fish showed a significantly improved health state (B I: p<10-16; B II: p<0.001) as indicated by externally visible changes of the skin, fins and gills. Specifically, reductions in haemorrhage of skin and fins (B II: p <0.05), and fin and gill necroses (B II: p<0.05), and gill inflammations (B II: p<0.02) were noted. Additionally, the sodium-humate treatment reduced integumental mucoses of skin, fins and gills (p<0.05) caused by Saprolegnia sp. The infection of skin and gills with protozoan parasites was reduced (p<0.01) as well. Ichthyophthirius sp. infections of the gills dropped significantly (p<0.05) in the treated fish groups as compared to the controls. Due to the observed mitigation of disease symptoms of environmental, bacterial, parasitic and fungal origin after a sodium-humate bath treatment of goldfish, a tissue protective, antiphlogistic, antimycotic and antiparasitic effect of sodium-humate can be concluded. For rearing of carp (F I) and rainbow trout (F III), 5 % Humocarb, formuliert� (type WH 67 A, Pharmawerk Weinb�hla) was mixed among the feed, while for the experiment F II 5 % Cellu-Ligno-Carbon-Isolat (type WH 67 G, Pharmawerk Weinb�hla) was added. In F I and F III a complete feed for trout was applied whereas for F II a complete feed for carp was used. In experiment F I (stocking density of both ponds 35,000 carp fry per ha) test fish revealed a higher mean body weight compared to the controls. With one exception (sample size to small), the difference between the groups was either significant (day 114: 65 vs. 45 g, p<0.05) or highly significant (day 1000: 1,146 versus 855 g, p<0.0001). The feed conversion ratio of the first experimental year (expurgated by HS-uptake) improved by 34 % due to HS application. Feed conversion ratio of the second and third year were falsified because of subjective feeding (which does not consider occurring losses). In experiment F II (stocking density of the 2 x 3 ponds (0,25 ha) were 20,000, 30,000 and 40,000 individuals per ha, respectively) no additional feed was given to the hatched fry during the first two weeks so that all the fish lived on natural feed items. Afterwards, bruised wheat grain was given until from day 63 either test or control feed was applied for a total of 87 days. At the end of the experiment, at the highest stocking density the mortality was significantly (p<0.0001) decreased by 20% in the test group supplied with HS. At low stocking density the total weight of fish at harvest was not different between the test and control group. However, at medium stocking density the average body weight was higher in the control whereas reverse conditions were observed at the highest stocking density (due to a loss of data no statistical analysis was possible). The feed conversion ratio (expurgated by the uptake of humic substances) was decreased in the test groups of low (5 %) and high (17 %) density. In both experiments (F I and F II) significantly reduced infections of the internal organs liver, kidney and spleen with potentially pathogenic Aeromonas and Pseudomonas spp. were detected. In the rainbow trout rearing (F III; N = 2 x 25), the HS-caused changes in consistency of feed and faeces impaired the formation of heavy algal films and large microorganism populations, and contributed to less turbid water turbidity in the aquaria, particularly at high temperatures. Significant differences in growth and feed conversion ratio could not be observed. In general, the addition of 5 % HS to the feed is regarded as being too high because the total amount of feed given (inclusive 5 % HS) is considered. The feed conversion ratio was higher in most of the HS-groups. Effects of oral application of 5 % HS in feed were thus less obvious than the bath treatment. Zootechnical parameters, in general, did not change. Merely in experiment F I the body weight of the HS-groups was significantly higher than that of the controls. In aquaria and small ponds used for ornamental fish breeding the observed cleaning of water and reduced formation of algal films and microorganism populations caused by HS could be of future importance. Intestinal effects of HS are assumed as internal organs revealed a reduced infection with both motile aeromonads and pseudomonads.

The objective of this project was the study of basic interaction processes in the systems actinide - clay organics - aquifer and actinide - natural clay - clay organics - aquifer. Thus, complexation, redox, sorption and diffusion studies were performed. To evaluate the influence of nitrogen, phosphorus and sulfur containing functional groups of humic acid (HA) on the complexation of actinides in comparison to carboxylic groups, the Am(III) and U(VI) complexation by model ligands was studied by UV-Vis spectroscopy and TRLFS. The results show that Am(III) is mainly coordinated via carboxylic groups, however, probably stabilized by nitrogen groups. The U(VI) complexation is dominated by carboxylic groups, whereas nitrogen and sulfur containing groups play a minor role. Phosphorus containing groups may contribute to the U(VI) complexation by HA, however, due to their low concentration in HA they play only a subordinate role compared to carboxylic groups. Applying synthetic HA with varying sulfur contents (0 to 6.9 wt.%), the role of sulfur functionalities of HA for the U(VI) complexation and Np(V) reduction was studied. The results have shown that sulfur functionalities can be involved in U(VI) humate complexation and act as redox-active sites in HA for the Np(V) reduction. However, due to the low content of sulfur in natural HA, its influence is less pronounced. In the presence of carbonate, the U(VI) complexation by HA was studied in the alkaline pH range by means of cryo-TRLFS (-120°C) and ATR FT-IR spectroscopy. The formation of the ternary UO2(CO3)2HA(II)4− complex was detected. The complex formation constant was determined with log β0.1 M = 24.57 ± 0.17. For aqueous U(VI) citrate and oxalate species, luminescence emission properties were determined by cryo-TRLFS and used to determine stability constants. The existing data base could be validated. The U(VI) complexation by lactate, studied in the temperature range 7 to 65°C, was found to be endothermic and entropy-driven. In contrast, the complex stability constants determined for U(VI) humate complexation at 20 and 40°C are comparable, however, decrease at 60°C. For aqueous U(IV) citrate, succinate, mandelate and glycolate species stability constants were determined. These ligands, especially citrate, increase solubility and mobility of U(IV) in solution due to complexation. The U(VI) sorption onto crushed Opalinus Clay (OPA, Mont Terri, Switzerland) was studied in the absence and presence of HA or low molecular weight organic acids, in dependence on temperature and CO2 presence using OPA pore water as background electrolyte. Distribution coefficients (Kd) were determined for the sorption of U(VI) and HA onto OPA with (0.0222 ± 0.0004) m3/kg and (0.129 ± 0.006) m3/kg, respectively. The U(VI) sorption is not influenced by HA (50 mg/L), however, decreased by low molecular weight organic acids (> 1×10-5 M), especially by citrate and tartrate. With increasing temperature, the U(VI) sorption increases both in the absence and in the presence of clay organics. The U(VI) diffusion in compacted OPA is not influenced by HA at 25 and 60°C. Predictions of the U(VI) diffusion show that an increase of the temperature to 60°C does not accelerate the migration of U(VI). With regard to uranium-containing waste, it is concluded that OPA is suitable as host rock for a future nuclear waste repository since OPA has a good retardation potential for U(VI).

In den letzten 15 Jahren wurde in einigen Mittelgebirgsregionen Zentraleuropas ein regional un-terschiedlich starker Anstieg der Konzentration gelösten organischen Kohlenstoffes (DOC) in Oberflächengewässern festgestellt. Besonders betroffen von dieser Entwicklung sind für die Trink-wasserproduktion genutzte Talsperren. Durch den erhöhten DOC-Gehalt wird die Trinkwasser-aufbereitung nicht nur kostenintensiver, sondern bekommt auch zunehmend Probleme bei der Wahrung der Trinkwasserqualität. Die Quellen des DOC sind in den Einzugsgebieten befindliche Moore und hydromorphe Nassstandorte. Die Ursachen für den in den vergangenen Jahren doku-mentierten Mehraustrag sind äußerst komplex und beinhalten sowohl klimatische als auch nut-zungsbedingte Veränderungen. Ziel der vorliegenden Arbeit ist, den Charakter des DOC näher zu bestimmen, um die für die Wasseraufbereitung problematischen Fraktionen zu benennen und Ansatzpunkte für regulierende Maßnahmen aufzuzeigen. Die Charakterisierung erfolgte an einem LC-OCD-System (Liquid Chromatography – Organic Carbon Detection) am Beispiel der drei Tal-sperrensysteme Muldenberg, Carlsfeld (Westerzgebirge) und Fláje (Osterzgebirge, Tschechien). Die Talsperren weisen unterschiedliche DOC-Gehalte auf. Die Ergebnisse zeigen, dass der DOC aller Untersuchungsgebiete sowohl in den Zuflüssen, dem Talsperrenwasser bzw. Rohwasser als auch dem Reinwasser der Wasserwerke von Huminstoffen dominiert wird. DOC-Schwankungen sind folglich auf Schwankungen der Huminstoffkonzentrationen zurückzuführen. In den Zuflüs-sen treten die höchsten Huminstoffkonzentrationen während der Schneeschmelze im Frühjahr und bei Starkniederschlägen im Sommer und Herbst auf. Niedrige Konzentrationen sind an Bo-dengefrornis oder Trockenphasen im Sommer gebunden. In den Talsperren tritt im Sommer eine Schichtung der Huminstoffe auf, bei der an der Oberfläche durch photolytischen Abbau niedrige-re Konzentrationen und in Abhängigkeit von der Talsperrentiefe durch die Einschichtung der Zuflüsse am Grund höhere Konzentrationen auftreten. Die Verteilung im Winter ist allein von der Einschichtung der Zuflüsse abhängig. Die Verteilung der Building Blocks (Huminstoffhydrolysate) ist im Sommer der der Huminstoffe entgegengesetzt, da sich beim photolytischen Abbau von Hu-minstoffen Building Blocks bilden. Während der Zirkulationen sind die Konzentrationen der Hu-minstofffraktionen im gesamten Tiefenprofil gleich. In den Talsperren Muldenberg und Carlsfeld wurde im Untersuchungszeitraum unter Berücksichtigung der technischen Gegebenheiten und anderer Wassergütekriterien der optimale Rohwasserentnahmehorizont (niedrigstmögliche Hu-minstoffkonzentration) gewählt. In der Talsperre Fláje könnte die Rohwasserqualität durch Instal-lation eines schwenkbaren Entnahmerohres erheblich verbessert werden. In den Wasserwerken sind die Huminstofffraktionen mit abnehmender Molekülgröße schwieriger zu entfernen. Die relativ niedermolekularen Building Blocks sind die am schwersten entfernbaren DOC-Bestandteile. Auf Grundlage mathematischer Beziehungen zu SAK254 bzw. DOC wurde ein Programm (DOCQuaC – DOC-Quality Calculator) entwickelt, mit dessen Hilfe die Huminstofffraktionen über diese Grundparameter ermittelt werden können. Neben der Fraktionsquantifizie-rung besteht im Programm die Möglichkeit zur Berechnung der Eliminierungsleistung, so dass beispielsweise nach der Durchführung einer Maßnahme zur Verbesserung der DOC-Entfernung eine unmittelbare Aussage über den Effekt auf die Fraktionen vorliegt. / For the last 15 years a change of Dissolved Organic Carbon (DOC) has been taking place in surface waters of Middle European Highlands. The degree of this increase is regionally dependent. The reasons as well as the consequences of this development are of high complexity. The changes are expressed in an increase of middle DOC values and, additionally, an increase of annual changes. Reservoirs, used for drinking water production, are particularly affected, because the heightened DOC content increases the costs for drinking water treatment, and currently it is not possible to maintain a good drinking water quality. The main DOC source are peat bogs, situated in the catchments. The cause of the increasing DOC output from peat bogs are very complex and contain changes as well in climate as in land use. The aim of this presented dissertation was to gain more information about the character of DOC, so that those fractions can be determined which are problematically to remove inside the waterworks. On that base it should be possible to find some starting points for more regulating measures. The analytical characterisation was carried out by the help of a LC-OCD-System (Liquid Chromatography – Organic Carbon Detection), exemplary for the reservoir systems Muldenberg, Carlsfeld (Western Ore Mountains) and Fláje (Eastern Ore Mountains, Czech Republic). Although the DOC concentrations are very different in the reser-voirs, it was always dominated by humic substances, namely as well in the tributaries, as in the raw and treated water. Thus changes in the DOC concentration are due to changes in the concen-tration of humic substances. In spring and summer occur the highest concentrations of humic substances in the tributaries. Low concentrations are linked with frost in winter or dry periods in summer. In the reservoirs in summer a stratification of humic substances shapes up, with lower concentration near the water surface (caused by photolytical degradation) and accordingly higher concentration near the ground (layering of influxes). Distribution in winter only depends on the layering of the influxes. In summer the concentration of Building Blocks (humic substance hydro-lysates) is opposite to that of humic substances, because of the creation of Building Blocks at the photolytical degradation. During circulation the concentration of all humic substance fractions are equal in each reservoir layer. In the reservoirs Muldenberg and Carlsfeld, a further optimisation of the raw water withdrawal horizon is not possible. In the reservoir Fláje the raw water could be much improved by installing a movable withdrawal device. With decreasing molecular size the elimination of the humic fractions gets more difficult in the waterworks. Most difficult to eliminate are Building Blocks. On the basis of the mathematical cor-relation between humic fractions and the SAC254 and DOC a program (DOCQuaC – DOC-Quality Calculator) has been developed, which enables die calculation of humic concentrations by enter-ing these general parameters. In addition to the calculation of the concentrations it is possible to calculate the elimination capacity for the humic fractions. Thus the effects of measures on the humic fractions, which could be done for optimisation of the elimination capacity, can be imme-diately observed.

Presented investigation and quantification of natural remediation of highly contaminated surface water systems located in South Ural via humic substances is the first step for sustainable developing of nuclear industry. In the surface water systems, humic substances are shown to promote the immobilization of radionuclides and decreasing of the bioavailability for fish contamination in the investigated water bodies. As proved in this thesis, the influence of humic substances on radionuclide sorption is provided by their special properties of a reversible transform into micelles. The theoretical approach based on consideration of protons as fermion gas in water solution was assumed and applied to this phenomenon, being due to the duel nature of humic substances molecules. The influence of humic substances is quantified as a modified Henry s low of sorption. Investigation of changing of electrostatical status of micelles with increasing of humic substances concentration in water solution leads to modelling of remediation effect of humic substances in respect to influence on fish in contaminated waters. This effect was interpreted and quantified, based on properties of proteins of gill s cell membranes under certain conditions in water solution. Humic substances appreciably influence the chemical and biological interactions between radionuclides and the environment that has experienced increasing interest concerning the remedial uses of humic materials.

This diploma thesis deals with the conformation and molecular organization of humic acids in aqueous solutions. Humic substances have heterogeneous and polydisperse nature, therefore their secondary chemical structure has not yet been defined properly, neither has their conformational arrangement. The conformation of humic substances in the solutions is mainly stabilized by weak disperse forces, such as Van der Waals, -, CH- interactions and hydrogen bonds. Humic substances in the solutions tend to form aggregates that are held together mostly by the intermolecular hydrophobic interactions. Concentration series of humic acids were prepared in three different mediums of constant ionic strength: NaOH and NaCl (prepared either by neutralization NaOH by HCl or direct dilution by solution of NaCl). Several analytical methods have been used to determine conformation and molecular organization of humic acids: potentiometric determination of pH, direct conductometry, ultraviolet and visible spectroscopy, density measurement, dynamic light scattering, laser Doppler velocimetry and high resolution ultrasound spectrometry. It was confirmed that the conformation and molecular organization of humic acids in aqueous solutions depend not only on pH of medium, but they also depend on chemical nature and concentration with the same pH. Results showed that hydrodynamic diameter of particles is significantly increasing in NaCl medium prepared by neutralization NaOH by HCl, particularly at low concentrations, which corresponds to higher values of zeta potential obtained. Concentration dependencies of ultrasonic velocity and compressibility also indicate the changes in conformation and molecular organization corresponding with results from other methods used.

Landfilling is the most widely used method for the disposal of municipal solid wastes (MSW) in the United States due to its simplicity and low cost. According to the 2014 report on Advancing Sustainable Materials Management by the USEPA, only 34% of the total MSW generated in the US was recycled, while 13% was combusted for energy recovery. In 2014, 53% of the MSW generated, (i.e. 136 million tons) in the US was landfilled. The treatment of landfill leachates, generated by percolation of water through the landfill, primarily due to precipitation, has been found to be one of the major challenges associated with landfill operation and management. Currently, leachates from most landfills are discharged into wastewater treatment plants, where they get treated along with domestic sewage. Issues associated with treatment of landfill leachates due to their high nitrogen and heavy metal content have been widely studied. Recently, it has been observed that the organic carbon in landfill leachates, specifically humic and fulvic acids (together referred to as "humic substances") contain aromatic groups that can absorb large amounts of ultraviolet (UV) light, greatly reducing the UV transmissivity in wastewater plants using UV disinfection as the final treatment step. This interference with UV disinfection is observed even when landfill leachates constitute a very small fraction (of the order of 1%) of the total volumetric flow into wastewater treatment plants. Humic substances are present as dissolved organic matter (DOM) and typically show very low biodegradability. Removing these substances using chemical treatment or membrane processes is an expensive proposition. However, the concentrations of humic substances are found to be reduced in leachates from landfill cells that have aged for several years, suggesting that these substances may be degraded under the conditions of long-term landfilling. The primary objective of this research was to use a two-stage process employing thermophilic pretreatment followed by a mesophilic anaerobic membrane bioreactor (AnMBR) to mimic the conditions of long-term landfilling. The AnMBR was designed to keep biomass inside the reactor and accelerate degradation of biologically recalcitrant organic carbon such as humic substances. The treatment goal was to reduce UV absorbance in raw landfill leachates, potentially providing landfills with an innovative on-site biological treatment option prior to discharging leachates into wastewater treatment plants. The system was operated over 14 months, during which time over 50% of UV-quenching organic carbon and 45% of UV absorbance was consistently removed. To the best of our knowledge, these removal values are higher than any reported using biological treatment in the literature. Comparative studies were also performed to evaluate the performance of this system in treating young leachates versus aged leachates. Next-generation DNA sequencing and quantitative PCR (qPCR) were used to characterize the microbial community in raw landfill leachates and the bioreactors treating landfill leachate. Analysis of microbial community structure and function revealed the presence of known degraders of humic substances in raw as well as treated landfill leachates. The total number of organisms in the bioreactors were found to be higher than in raw leachate. Gene markers corresponding to pathogenic bacteria and a variety of antibiotic resistance genes (ARGs) were detected in raw landfill leachates and the also in the reactors treating leachate, which makes it necessary to compare these ARG levels with wastewater treatment in order to determine if leachates can act as sources of ARG addition into wastewater treatment plants. In addition, the high UV absorbance of leachates could hinder the removal of ARBs and ARGs by UV disinfection, allowing their release into surface water bodies and aiding their proliferation in natural and engineered systems. / Ph. D.

The diploma thesis investigates the coagulation of nature waters with humic substances and peptides/proteins produced by Microcystis aeruginosa during water treatment process with aluminum coagulants. It was confirmed that the efficiency of coagulation mechanism of humic substances and peptides/proteins strongly depends on the pH value, because pH value is limited factor of a charge properties of peptides/proteins, humic substances and hyrolysis products of coagulants. The optimal pH range for the coagulation of humic substances was 5- 6 and for cyanobacterial peptides/proteins 5-6.5. In both cases a charge neutralisation and an adsorption were dominant mechanism of coagulation. Optimal pH range for the coagulation of humic substances in the presence of cyanobacterial peptides/proteins was identical as well as optimal pH range of humic substances with absence of peptides/proteins (5-6) but the presence of peptides/proteins positively influenced particle coagulation of humic substances, because the optimal concentration of the coagulants decreased to more than half. An interaction between humic substances and peptides/proteins was demonstrated. A positive effect of coagulation was observed in the case of jar tests with absence of coagulants at a very low pH range (<4). Keywords Humic substances; AOM...

A produção de mudas via propagação vegetativa é fundamental para se obter os ganhos desejados nos programas de melhoramento genético, bem como garantir o sucesso da implantação florestal. No entanto, existem muitos fatores que impedem a sua utilização de forma abrangente na produção de mudas para algumas espécies florestais, principalmente em relação à baixa taxa de enraizamento. Para atender a essa necessidade, pesquisas com o uso de substâncias que promovam maior taxa de enraizamento de miniestacas e crescimento de mudas vem sendo desenvolvidas, como o uso de reguladores de crescimento e de bioestimulantes à base de substâncias húmicas. Dessa forma o objetivo do presente trabalho é avaliar o efeito de adubos foliares e condicionadores de solo na produção de mudas clonais de híbridos de Pinus caribaea var. hondurensis vs. elliottii var. elliottii e de Pinus tecunumanii vs. caribaea var. hondurensis. Os experimentos foram desenvolvidos em viveiro florestal na cidade de Paranapanema-SP, com delineamento de blocos ao acaso, em arranjo fatorial 4x2, com duração de seis meses. Foram utilizados dois condicionadores de solo, sendo o C1 à base de pescados marinhos e o C2 à base de ácidos húmicos e quatro adubos foliares, sendo AF1 à base de quitosana, AF2 à base de pescados marinhos, AF3 à base de ácido húmico e AF4 extrato da alga Ascoplhyllum nodosum, além da mistura dos adubos M1AF (AF2+AF3+AF4) e o M2AF (AF1+AF3+AF4). Foram instalados dois experimentos: 1) híbrido de P. caribaea var. hondurensis vs. elliottii var. elliottii, testando a combinação dos dois condicionadores de solo e os adubos AF2, AF3, AF4 e M1AF e 2) híbrido de P. tecunumanii vs. carearia var. hondurensis em que a única alteração nos tratamentos foi o AF2 por AF1 e o M1AF por M2AF. As mudas foram avaliadas nas três fases da seguinte forma: 1) casa de vegetação: taxa de enraizamento; 2) casa de sombra: altura, massa seca e verde da parte aérea e radicial e a relação entre elas; e 3) rustificação: avaliadas as mesmas variáveis da casa de sombra, além do estado nutricional das acículas. Para o experimento 1, a taxa de enraizamento com C2 + AF3 foi 46,67% maior quando comparada à testemunha, comprovando o efeito positivo das substâncias húmicas no enraizamento. Já na fase de rustificação, a interação C1 + AF3 promoveu maior crescimento das mudas. Para o experimento 2, não houve diferença nas taxas de enraizamento na casa de vegetação, devido a característica de rápido crescimento do P. tecunumanii. E, na fase de rustificação, o adubo foliar AF4 promoveu maior crescimento somente em altura. Na casa de sombra, os tratamentos testados não promoveram crescimento significativo para ambos os híbridos. Os teores nutricionais analisados nas acículas estão dentro dos níveis adequados para ambos os experimentos. Somente o Mn apresentou baixo teor, mas sem sintomas de deficiência no experimento 2. Os híbridos obtiveram respostas diferentes quando submetidos a tratamentos semelhantes e a determinação do tratamento adequado vai depender do gargalo na produção de mudas, podendo este ser o enraizamento ou o crescimento. / The production of cuttings by vegetative propagation is essential to obtain the desired accomplishment in genetic improvement programs and ensure the success of forest establishment. However there are many factors that prevent their use in a comprehensive manner considering the production of cuttings for some forest species especially in low rooting rate. In order to meet this need research on the use of substances that promote greater rooting rates of minicuttings and growth of cuttings have been developed with the use of growth regulators and biostimulants based on humic substances. Thus the purpose of this study is to evaluate the effect of foliar fertilizers and soil conditioners in the production of Pinus caribaea var. hondurensis vs. elliottii var. elliottii and Pinus tecunumanii vs. caribaea var. hondurensis hybrids cuttings. The experiment was conducted in the city of Paranapanema-SP using cutting nursery with delineation of randomized blocks in a factorial arrangement 4x2, lasting six months. It used two soil conditioners, the C1 based on marine fish and C2 based on humic acids and 4 foliar fertilizers, the AF1 chitosan base, AF2 based on marine fish, AF3 based on humic acid and AF4 by Ascoplhyllum nodosum seaweed extract, besides the mixture of fertilizers M1AF (AF2;AF3;AF4) and M2AF (AF1;AF3;AF4). Two experiments were implemented: 1) hybrid of P. caribaea var. hondurensis vs. elliottii var. elliottii, testing the combination of two soil conditioners as well as fertilizers AF2, AF3, AF4 and M1AF and 2) hybrid of P. tecunumanii vs. caribaea var. hondurensis in which the only change in treatment was the AF2 by AF1 and M1AF by M2AF. Cuttings were evaluated in three phases as follows: 1) greenhouse: rooting rate; 2) shade house: higher, aerial drywet mass and radicial part as well as the relationship between them; 3) hardening: the shade house had the same variables evaluated, in addition with the nutritional status of needles. For the experiment 1, the rooting rate with C2 + AF3 were 46.67% higher compared with the control, this stated the positive effect of humic substances on rooting. Considering the hardening phase C1 + AF3 interaction promoted greater seedling growth. For the experiment 2 there was no difference in rooting rates at greenhouse, because the characteristic of fast growth of P. tecunumanii. In hardening phase, the foliar fertilizer AF4 promoted greater growth only in height. In the shade house treatments did not promote significant growth for both hybrids. Nutritional contents analyzed the needles leaves are within appropriate levels for both experiments. Mn showed only low level, but without deficiency symptoms in the experiment 2. Different answers were obtained on hybrids when subjected to similar treatment and to determine the appropriate treatment will depend on the obstaclecuttings production, which may either rooting or growth.

O presente trabalho teve como objetivo avaliar a remoção de três concentrações diferentes de microcistina extracelular em diferentes combinações de tratamento de águas para abastecimento, em escala de bancada, que tiveram como sequência básica a clarificação associada ou não aos processos de pré-oxidação com \'K\'MN\'O IND.4\', adsorção em CAP e pós-cloração. Os resultados mostraram que para todas as águas estudadas o permanganato de potássio não interferiu nos mecanismos de coagulação/floculação e ainda mostrou-se uma alternativa segura para realização da pré-oxidação no que tange à formação de THMs. Na Fase 1, com concentração inicial de microcistina extracelular em torno de 1,4 \'mü\'g/L, a clarificação (coagulação, floculação, flotação por ar dissolvido e clarificação final) atendeu ao padrão de potabilidade que determina concentrações de microcistina menores que 1,0 \'mü\'g/L. Já na Fase 2, com concentração inicial microcistina extracelular em torno de 21,7 \'mü\'g/L, para o atendimento à legislação foi necessário associar a clarificação à pré-oxidação, dosando-se 1,0 ou 2,0 mg \'K\'MN\'O IND.4\'/L, e à pós-cloração com 3,0 mg \'CL IND.2\'/L. Na Fase 3, com concentração inicial de microcistina extracelular em torno de 64,1 \'mü\'g/L, a associação da clarificação com a adsorção com 60,0 mg/L de CAP e com a pós-cloração com 3,0 mg \'CL IND.2\'/L proporcionou residuais de microcistina extracelular inferiores à 1,0 \'mü\'g/L. Observou-se ainda, que nas Fases 1 e 3 a presença de matéria orgânica dissolvida interferiu negativamente nas sequências de tratamento ao consumir parte do permanganato de potássio destinado à oxidação da microcistina extracelular. Entretanto, na Fase 2 a demanda do pré-oxidante pelas substâncias húmicas parece ter impedido a lise de parte das células de Microcystis sp. / The present work had as objective to evaluate the removal of three different concentrations of extracellular microcystins in different combinations of water treatment for supplying, in bench scale, that had as basic sequence the clarification associated or not with the processes of pre-oxidation with \'K\'MN\'O IND.4\', adsorption on PAC and post-chlorination. The results showed that for all waters studied the potassium permanganate did not interfere in the mechanisms of coagulation/flocculation and also proved to be a safe alternative for achieving the pre-oxidation with regard to the formation of THMs. In Phase 1, with initial concentration of extracellular microcystin around 1.4 \'mü\'g/L, the clarification (coagulation, flocculation, dissolved air flotation and clarification final) met the World Health Organization drinking water guideline value of 1.0 \'mü\'g/L of microcystin. Already, in Phase 2, with initial concentration extracellular microcystin around 21.7 \'mü\'g/L, to meet the legislation was necessary to involved the clarification with the pre-oxidation, dosing 1.0 or 2.0 mg \'K\'MN\'O IND.4\'/L, and with the post-chlorination with 3.0 mg \'CL IND.2\'/L. In Phase 3, with initial concentration of extracellular microcystin around 64.1 \'mü\'g/L, the association of clarification with the adsorption with 60.0 mg/L of PAC and the post-chlorination with 3.0 mg\'CL IND.2\'/L provided residual extracellular microcystin below 1.0 \'mü\'g/L. It was also observed that in Phases 1 and 3 the presence of dissolved organic matter intervened negatively in the sequence of treatment when consuming part of the potassium permanganate destined to the oxidation of extracellular microcystin. However, in Phase 2 the demand for pre-oxidizing by the humic substances seems to have prevented the lysis of some cells of Microcystis sp.