Submitted by Leticia Schettini (leticia@ufrrj.br) on 2016-09-21T15:12:59Z No. of bitstreams: 1 2011 - Camila Pinheiro Nobre.pdf: 2714138 bytes, checksum: bc436d879ddfded703d4d5fea9f9943c (MD5) / Made available in DSpace on 2016-09-21T15:12:59Z (GMT). No. of bitstreams: 1 2011 - Camila Pinheiro Nobre.pdf: 2714138 bytes, checksum: bc436d879ddfded703d4d5fea9f9943c (MD5) Previous issue date: 2011-02-17 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior-CAPES / The aim of this study was to observe the germination, production of glomalin and monitor development of species of mycorrhizal fungi (AMF) of the germplasm bank of Embrapa in root organ culture (ROC) of basil and bryophytes in vitro, analyzing their interaction with the hosts and the influence of the culture medium enriched with humic acids on fungal growth and bryophyte Lunularia cruciata. For this, some AMF species were selected and had their glomerospores extracted and subjected to surface disinfection process, placed in water-agar medium and temperature-controlled chamber to germinate. A germination test was conducted for 15 days, and the results were analyzed by ANOVA and Tukey test applied to 5% probability. Species with germinated glomerospores (Gigaspora margarita, Glomus manihots, Scutellospora heterogama and Glomus proliferum) were placed in ROC of purple basil where they had their growth observed until 100 days after inoculation. Also as part of the characterization of AMF species it was quantified the level of glomalin in the samples of multiplication and the results were subjected to analysis of variance and Scott-Knott test at 5% probability. In the second chapter it was investigated the effect of mycorrhizal association in ROC of purple basil, and in the third chapter the influence of different concentrations of humic acid and association with growth of Lunularia cruciata (area and length). The results were submitted to ANOVA and Tukey test at 5% probability. Scutellospora heterogama was the species with higher germination rates of glomerosporos, followed by Gigaspora margarita. The species of Glomus sporulated after formation of symbiosis. The amount of glomalin produced by different AMF was distinct, especially in total glomalin fraction. Different AMF species did not show difference in efficiency to promote development of Ocimum basilicum transformed roots. The growth of basil transformed roots in the MSR was extended from 15 days after inoculation with mycorrhizal fungi. The usage of humic acids in the culture medium in concentrations of 20 and 80 mg CL-1 enhanced growth of bryophyte L. cruciata, and its association with mycorrhizal fungi, as well as promoted the highest number of spores of Gl. proliferum. The association L. cruciata and AMF was characterized as mutualistic, since both had advantages in growth and sporulation. Gigaspora margarita and Glomus proliferum increased growth of Lunularia cruciata. / O objetivo do trabalho foi observar a germina??o e produ??o de glomalina e acompanhar desenvolvimento de esp?cies de fungos micorr?zicos arbusculares (FMA) do banco de germoplasma da Embrapa em ra?zes geneticamente modificadas de manjeric?o e em bri?fitas in vitro. Ainda, avaliar sua intera??o com os hospedeiros e a influ?ncia de meio de cultura enriquecido com ?cidos h?micos no crescimento do fungo e da bri?fita Lunularia cruciata. Para isso algumas esp?cies de FMAs foram selecionadas e tiveram seus glomerosporos extra?dos e submetidos ao processo de desinfesta??o superficial, colocados em meio Agar?gua e c?mara com temperatura controlada para germinar. Realizou-se teste de germina??o por 15 dias e os resultados foram submetidos a an?lise de vari?ncia e aplicado teste de Tukey ? 5% de probabilidade. Esp?cies com glomerosporos germinados (Gigaspora margarita, Glomus manihots, Scutellospora heterogama e Glomus proliferum) foram colocadas em ra?zes modificadas de manjeric?o roxo onde tiveram seu crescimento observado at? 100 dias ap?s a inocula??o. Ainda como parte da caracteriza??o de esp?cies de FMAs foi realizado a quantifica??o dos teores de glomalina nas amostras de multiplica??o sendo os resultados submetidos a an?lise de vari?ncia e aplicado teste de Scott-Knott ? 5% de probabilidade. No segundo cap?tulo foi verificado o efeito da associa??o FMAs em ra?zes modificadas de manjeric?o roxo e no terceiro cap?tulo a influ?ncia da associa??o ?cido h?mico em diferentes concentra??es, bri?fita Lunularia cruciata (?rea e comprimento) e FMAs. Os resultados foram submetidos ? an?lise de vari?ncia e teste de Tukey a 5% de probabilidade. Scutellospora heterogama foi a esp?cie com maiores taxas de germina??o de glomerosporos, seguida da Gigaspora margarita. As esp?cies de Glomus esporularam logo ap?s a forma??o da simbiose. A quantidade de glomalina produzida pelos diferentes FMAs foi distinta, em especial na fra??o glomalina total. As diferentes esp?cies de FMAs n?o apresentaram distin??o na efici?ncia de promover o desenvolvimento das ra?zes transformadas de Ocimum basilicum. O crescimento de ra?zes transformadas de manjeric?o em meio MSR foi ampliado a partir dos 15 dias ap?s a inocula??o de fungos micorr?zicos. O uso de ?cidos h?micos no meio de cultura em concentra??es de 20 e 80 mg C.L-1 incrementou o crescimento da bri?fita Lunularia cruciata e sua associa??o com fungos micorr?zicos arbusculares, assim como promoveram a maior esporula??o de Gl. proliferum. A associa??o Lunularia cruciata e FMAs foi caracterizada como mutualista j? que ambos apresentaram benef?cios em crescimento e esporula??o. Gigaspora margarita e Glomus proliferum promoveram maior crescimento de Lunularia cruciata.

Immobilisierung bzw. Mobilisierung und Transport von Schadstoffen in der Umwelt, besonders in den Kompartimenten Boden und Wasser, sind von fundamentaler Bedeutung für unser (Über)Leben auf der Erde. Einer der Hauptreaktionspartner für organische und anorganische Schadstoffe (Xenobiotika) in der Umwelt sind Huminstoffe (HS). HS sind Abbauprodukte pflanzlichen und tierischen Gewebes, die durch eine Kombination von chemischen und biologischen Ab- und Umbauprozessen entstehen. Bedingt durch ihre Genese stellen HS außerordentlich heterogene Stoffsysteme dar, die eine Palette von verschiedenartigen Wechselwirkungen mit Schadstoffen zeigen. Die Untersuchung der fundamentalen Wechselwirkungsmechanismen stellt ebenso wie deren quantitative Beschreibung höchste Anforderungen an die Untersuchungsmethoden. Zur qualitativen und quantitativen Charakterisierung der Wechselwirkungen zwischen HS und Xenobiotika werden demnach analytische Methoden benötigt, die bei der Untersuchung von extrem heterogenen Systemen aussagekräftige Daten zu liefern vermögen. Besonders spektroskopische Verfahren, wie z.B. lumineszenz-basierte Verfahren, besitzen neben der hervorragenden Selektivität und Sensitivität, auch eine Multidimensionalität (bei der Lumineszenz sind es die Beobachtungsgrößen Intensität IF, Anregungswellenlänge lex, Emissionswellenlänge lem und Fluoreszenzabklingzeit tF), die es gestattet, auch heterogene Systeme wie HS direkt zu untersuchen. Zur Charakterisierung können sowohl die intrinsischen Fluoreszenzeigenschaften der HS als auch die von speziell eingeführten Lumineszenzsonden verwendet werden. In beiden Fällen werden die zu Grunde liegenden fundamentalen Konzepte der Wechselwirkungen von HS mit Xenobiotika untersucht und charakterisiert.<br> Für die intrinsische Fluoreszenz der HS konnte gezeigt werden, dass neben molekularen Strukturen besonders die Verknüpfung der Fluorophore im Gesamt-HS-Molekül von Bedeutung ist. Konformative Freiheit und die Nachbarschaft zu als Energieakzeptor fungierenden HS-eigenen Gruppen sind wichtige Komponenten für die Charakteristik der HS-Fluoreszenz. Die Löschung der intrinsischen Fluoreszenz durch Metallkomplexierung ist demnach auch das Resultat der veränderten konformativen Freiheit der HS durch die gebundenen Metallionen. Es zeigte sich, dass abhängig vom Metallion sowohl Löschung als auch Verstärkung der intrinsischen HS-Fluoreszenz beobachtet werden kann.<br> Als extrinsische Lumineszenzsonden mit wohl-charakterisierten photophysikalischen Eigenschaften wurden polyzyklische aromatische Kohlenwasserstoffe und Lanthanoid-Ionen eingesetzt. Durch Untersuchungen bei sehr niedrigen Temperaturen (10 K) konnte erstmals die Mikroumgebung von an HS gebundenen hydrophoben Xenobiotika untersucht werden. Im Vergleich mit Raumtemperaturexperimenten konnte gezeigt werden, dass hydrophobe Xenobiotika an HS-gebunden in einer Mikroumgebung, die in ihrer Polarität analog zu kurzkettigen Alkoholen ist, vorliegen. Für den Fall der Metallkomplexierung wurden Energietransferprozesse zwischen HS und Lanthanoidionen bzw. zwischen verschiedenen, gebundenen Lanthanoidionen untersucht. Basierend auf diesen Messungen können Aussagen über die beteiligten elektronischen Zustände der HS einerseits und Entfernungen von Metallbindungsstellen in HS selbst angeben werden. Es ist dabei zu beachten, dass die Experimente in Lösung bei realen Konzentrationen durchgeführt wurden. Aus Messung der Energietransferraten können direkte Aussagen über Konformationsänderungen bzw. Aggregationsprozesse von HS abgeleitet werden. / Transport and fate of xenobiotics in the environment, especially in water and soil, are of utmost importance for life on earth. A major reaction partner for xenobiotics in the environment are humic substances (HS). HS are degradation products of plant and animal tissue, which are formed in a combination of subsequent chemical and/or biochemical processes. Because of the complex history of their origin HS are extremely heterogeneous mixtures of different compounds. Consequently, they posses a great variety of interaction capabilities with various xenobiotics. The investigations of the fundamental interaction mechanisms between HS and xenobiotics make high demands on the analytical techniques used. Especially spectroscopic techniques are promising for the investigation of interaction mechanisms in complex systems. Luminescence spectroscopy has the great advantage of outstanding sensitivity and of multidimensionality, which in principle allows the investigation of HS under environmental relevant conditions. For the characterisation of interaction processes of HS with xenobiotics the intrinsic fluorescence of HS as well as the luminescence of extrinsic probes can be used.<br> The intrinsic HS fluorescence is determined by the molecular structure as well as the connection of the basic fluorophores. Conformational freedom as well as the presence of energy accepting groups in the neighbourhood of the fluorophores are highly important for the overall intrinsic HS fluorescence. The presence of metal ions can either quench or enhance the intrinsic HS fluorescence, which depends on the metal ion as well as on the origin of the HS investigated. While in most cases Al3+ ions enhance the intrinsic HS fluorescence, Ln3+ ions induce a fluorescence quenching.<br> Polycyclic aromatic hydrocarbons were used as extrinsic fluorescence probes in order to characterize the interaction of HS and hydrophobic organic xenobiotics. In investigations at ultra-low temperatures (10 K) it could be shown that pyrene is bound in a HS microenvironment with an polarity which resembles that of small alcohols (e.g., butanol).<br> In case of metal complexation, the lanthanide ions Eu3+ and Tb3+ were used as luminescence probes. Due to the outstanding luminescence properties of those ions, information about metal binding sites in HS were obtained. Based on the measurements of intramolecular and intermolecular energy transfer processes average distances of metal binding sites were deduced.

Many chemicals present in the aquatic environment can interfere with physiological functions in fish. Exposure to chemicals can be revealed by the use of biomarkers. Induction of 7-ethoxyresorufin O-deethylase (EROD) activity is a commonly used biomarker for exposure to CYP1A inducers such as dioxins and polyaromatic hyrdrocarbons. Vitellogenin is a frequently used biomarker for estrogenic compounds in various fish species whereas a biomarker for androgens, spiggin, is only found in sticklebacks. The main objectives of this thesis were to evaluate gill EROD activity as a biomarker and the three-spined stickleback as a model species in ecotoxicological studies. EROD activities were measured in gill, liver and kidney in rainbow trout (Oncorhynchus mykiss) caged in urban areas in Sweden. EROD induction was most pronounced in the gill. Also in fish caged at reference sites, with an expected low level of known CYP1A inducers, a marked gill EROD induction was found. One suggested inducer in rural waters is humic substances (HS). To evaluate the EROD-inducing capacity of HS, three-spined sticklebacks (Gasterosteus aculeatus) were exposed to HS of natural or synthetic origin. Both kinds of HS caused significant EROD induction. Gill EROD activities were also induced in sticklebacks exposed to ethynylestradiol (EE2) and β-naphthoflavone (βNF), alone and in combinations. Production of vitellogenin was induced in sticklebacks exposed to ≥50 ng EE2/l and a significant decrease in spiggin production was observed in individuals exposed to 170 ng EE2/l. Results from this thesis further strengthen the contention that gill EROD activity is a very sensitive biomarker for CYP1A inducers and that the stickleback is a suitable biomonitoring species, especially for exposure to CYP1A inducers. The finding that not only classical CYP1A inducers but also HS and high EE2 concentrations stimulate gill EROD activity is of significance for the interpretation of biomonitoring data.

Objective of this project was the study of interaction processes between humic substances, U(VI), Np(V) and kaolinite KGa-1b. It contributed to the attainment of a better process understanding, the improvement of the knowledge on the interaction of humic substances and metal ions and the enhancement of the thermodynamic database. With a synthetic humic acid (HA), N-containing functional groups of HA were characterized by 15N-NMR spectroscopy. Based on these results, model studies of the influence of amino groups on the complexation behavior of HA were performed. Spectroscopic studies with amino acids show that the amino group do not contribute to the U(VI) complexation at pH 4. The impact of kaolinite on the formation of HA and humic substance-kaolinite-sorbates was studied in model syntheses. The results exhibit that the presence of kaolinite during the syntheses mainly influences the yields on HA and their elemental compositions. Synthetic humic substance-kaolinite-sorbates were isolated. Under exclusion of CO2, the U(VI) complexation by HA was investigated at pH 7 by means of the conventional time-resolved laser-induced fluorescence spectroscopy (TRLFS) and TRLFS with ultrafast pulses. Complexation parameters for the ternary complex UO2(OH)HA(I) were determined. Studies of the Np(V) reduction in presence of HA with different functionalities under anaerobic conditions have shown that Np(V) is reduced to Np(IV) by HA. The redox capacity depends on the HA functionality. Applying a modified HA it was verified that phenolic/acidic OH groups play a dominating role in the Np(V) reduction. The influence of HA on the U(VI) and Np(V) sorption onto kaolinite was investigated in batch experiments. In dependence on the experimental conditions, HA effects the sorption and consequently the mobility of U(VI) and Np(V). From studies of the U(VI) sorption onto synthetic humic substance-kaolinite-sorbates it was concluded that the structure and functionality of sorbed/associated humic substances considerably influence the sorption behavior of U(VI). The structure of U(VI)-kaolinite-surface complexes in presence of HA was studied by means of X-ray absorption spectroscopy and TRLFS and compared to those of U(VI)-kaolinite-complexes. Investigations of the migration of HA and U(VI) in the laboratory system kaolinite-water were carried out in diffusion experiments. The migration of HA in compacted clay is governed by diffusion and influenced by its colloidal properties. Humic substances exert an immobilizing effect on the U(VI) transport in compacted kaolinite.

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.

O líquido denominado “lixiviado”, formado pela disposição de resíduos em aterros sanitários e lixões, apresenta composição complexa, dada principalmente pela presença de compostos orgânicos extremamente recalcitrantes. O tratamento consorciado deste efluente com esgotos domésticos em ETE’s municipais carece de estudos, tendo em vista que são líquidos com características diferentes. Neste trabalho foi avaliado o desempenho de um processo biológico de tratamento de efluentes, composto por 3 sistemas de banhados construídos (B1, B2 e B3) operados em paralelo, tratando a combinação de esgoto doméstico (90%) + lixiviado de aterro sanitário (10%), no que diz respeito a remoção de matéria orgânica de lenta degradação. Cada sistema foi composto por 4 leitos cultivados com as macrófitas emergentes Luziola peruviana e Typha latifolia, recebendo afluente proveniente de diferentes níveis de pré-tratamento, nomeadamente: B1 - processo biológico anaeróbio (UASB) seguido de aeróbio (CBR); B2 - processo biológico aneróbio (UASB) e B3 - sem prévio tratamento. Para tal, além das análises da matéria orgânica pelos parâmetros não específicos, foi também empregada análise de absorbância e fluorescência para analisar a presença de substâncias húmicas e o uso da respirometria para avaliação da DQO lentamente biodegradável. As eficiências médias de remoção dos sistemas B1, B2 e B3 foi de 97,4, 96,1 e 96,6 % para DBO5, 66,9, 70,9 e 80,6% para DQOT, 61,9, 69 e 62,8 % para DQOFil e de 60,3, 72,8 e 72 % para COT, respectivamente. A avaliação destes parâmetros permitiu inferir que o nível de pré-tratamento não influenciou na eficiência de remoção pelos sistemas de banhados. Os espectros de absorbância foram marcados por maior absorbância na região λ < 400 nm, com diminuição progressiva até 700 nm. A aplicação de coeficientes de absortividade indicaram a predominância de matéria orgânica dissolvida proveniente de esgoto doméstico, com pouca influência de SH’s. Nos espectros de emissão de fluorescência, o aparecimento de bandas na região de 450 nm indicou a presença de substâncias húmicas no afluente e efluente do sistema de banhados construídos, sem diferença significativa entre a intensidade de fluorescência emitida em 450 nm nas amostras analisadas, indicando que a substância em questão passou pelo tratamento sem ser degradada. Os resultados observados pela análise de fluorescência sincronizada do B1 apresentaram contribuição positiva do pré-tratamento (menor concentração de DQOFil) para o desempenho do banhado, devido a menor presença de ácidos húmicos no efluente. / The liquid known as "leachate", produced by the disposal of waste in landfills and dumps, has a complex composition, characterized mainly by the presence of recalcitrant organic compounds. Currently, there is a lack of knowledge regarding the co-treatment of leachate with municipal wastewater in WWTP's, considering that these are effluents with different characteristics. This study evaluated the performance of a biological treatment process of wastewater consisting of 3 constructed wetlands systems (B1, B2 and B3), operated in parallel, treating a combination of domestic sewage (90%) + landfill leachate (10 %). Each system was comprised of four beds cultured with emerging macrophytes Luziola peruviana and Typha latifolia receiving affluent from different levels of pre-treatment, namely: B1 - anaerobic biological process (UASB) followed by aerobic biological process (CBR); B2 - anaerobic biological process (UASB). B3 received the untreated effluents. The content of organic matter and humic substances was analyzed by non-specific parameters, absorbance and fluorescence methods. Respirometry test were conducted to evaluate the content of slowly biodegradable COD. The average removal efficiencies of systems B1, B2 and B3 was of 97.4, 96.1 and 96.6% for BOD5, 66.9, 70.9 and 80.6% for CODT, 61,9, 69 and 62.8% for CODFil and 60.3, 72.8 and 72% of TOC, respectively. The evaluation of these parameters allowed to infer that the level of pre-treatment did not influenced the removal efficiency by wetland systems. Absorbance spectra were marked by greater absorbance in the region λ = <400 nm with progressive decrease up to 700 nm. Absorptivity coefficients indicated the predominance of domestic sewage, with little influence of SH's. In the fluorescence emission spectra, the appearance of bands in the 450 nm region indicated the presence of humic substances in the influent and effluent of the constructed wetland systems without a significant. This indicates that the substance in question went through the treatment without being degraded. The results obtained by synchronous fluorescence analysis showed a positive contribution of the B1 pretreatment (lowest concentration DQOFil) to the observed performance due to less presence of humic acids in the effluent. The authors did not have success during evaluation of the slowly biodegradable COD using the method proposed before.

Made available in DSpace on 2014-06-11T19:33:38Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-06-27Bitstream added on 2014-06-13T20:05:52Z : No. of bitstreams: 1 marcelo_av_dr_jabo.pdf: 1194015 bytes, checksum: 68c9171ad52d863ef75fe8bb9b03c925 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Em sistema de semeadura direta, os resíduos das culturas de entressafra são utilizados para protegerem a superfície do solo dos agentes erosivos e promoverem a ciclagem de nutrientes. O objetivo desse trabalho foi avaliar o efeito de sequências de culturas na quantidade, qualidade, decomposição e liberação de nutrientes dos resíduos vegetais de culturas de entressafra, bem como a cobertura do solo proporcionada pelos resíduos, na fertilidade do solo, nas frações particulada e associada aos minerais de C, nas substâncias húmicas do solo e na produtividade de milho e soja cultivados no verão, em região de clima tropical. O experimento foi conduzido em Jaboticabal, SP (48°15’22’’ W e 21°18’58’’ S), em um Latossolo Vermelho eutrófico. O delineamento experimental foi em faixas, com três repetições. Os tratamentos foram constituídos pela combinação de três sequências de culturas de verão (rotação soja-milho, monocultura de milho e monocultura de soja) com sete culturas de entressafra (milho, sorgo, girassol, crotalária, guandu, nabo forrageiro e milheto). O experimento foi iniciado em 2002 e o presente estudo se refere aos anos agrícolas 2008/2009 e 2009/2010. Avaliaram-se as quantidades de matéria seca e acúmulo de N, P, K, Ca, Mg e S pelas culturas de entressafra, bem como a dinâmica de decomposição e liberação dos nutrientes, por meio da utilização de sacolas de decomposição, com os seguintes períodos de avaliação: 15, 30, 60, 120 e 180 dias. A cobertura do solo foi avaliada após o manejo e ao final de cada ano agrícola. Amostras de solo foram coletadas nas camadas 0-10, 10-20 e 20-30 cm de profundidade em duas ocasiões, antes da semeadura das culturas de entressafra, em março, e de verão, em outubro, nos anos de 2008 e 2009. As amostras de solo foram submetidas às análises químicas para determinação dos teores... / In no-tillage system, the offseason crops residues are expected to maintain the soil covered in order to control soil erosion and to promote the nutrients cycling. The objective of this study was to evaluate the offseason crops dry matter quality and production, soil cover, residues decomposition and nutrient release and their effects on soil fertility, C content of humic substances, particulate and mineral associated organic matter fractions and summer maize and soybean yield. A field experiment has been carried out on a Rhodic Eutrudox at Jaboticabal, SP, Brazil (48°15’22’’ W and 21°18’58’’ S). A randomized split-block design with three replications was used. The treatments were the combination of three summer crops sequences (soybean-corn rotation, monoculture of corn and monoculture of soybean) with seven offseason crops (maize, grain sorghum, sunflower, sunn hemp, pigeon pea, oilseed radish and pearl millet). The cultivations started in 2002 and this study is related to 2008/2009 and 2009/2010 growing seasons. The offseason crops dry matter production and N, P, K, Ca, Mg and S accumulations were evaluated at management moment. The period of time to measure the residue decomposition and the nutrient release was 15, 30, 60, 120 and 180 days, using litter bags. The soil cover was evaluated after the offseason crops management and at the end of each growing season. The soil was sampled at 0-10, 10-20 and 20-30 cm soil depths in two occasions, before the offseason crops sowing, at March, and before the summer crops sowing, at October, in 2008 and 2009. Total organic C, pH, P, K, Ca, Mg and H+Al were determined in each soil samples. The soil samples obtained on October at 0-10 cm were submitted for physical and chemical organic matter fractionation, for particulate, mineral-associated, humic acid, fulvic acid and humin C content determination. The summer monoculture... (Complete abstract click electronic access below)

Made available in DSpace on 2014-06-11T19:35:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-05-19Bitstream added on 2014-06-13T18:46:19Z : No. of bitstreams: 1 bellin_ic_dr_araiq.pdf: 1668200 bytes, checksum: bca88ddf4d8550b63d36e4e2d483f6cb (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Neste trabalho caracterizaram-se amostras de solo e substâncias húmicas (SH) extraídas de diferentes solos da Bacia do Médio Rio Negro e também determinaram-se a capacidade de complexação de íons Hg(II) por SH e constantes de troca entre espécies metálicas originalmente complexadas às SH por íons Hg(II). As amostras de solos foram caracterizadas através do teor de matéria orgânica, análises elementar e granulométrica. Os resultados mostraram que os teores de matéria orgânica para as amostras de solo diminuíram com a profundidade. Com base nos resultados de análise granulométrica feita nas amostras de solos, caracterizou-se o aumento dos teores de argila em função da profundidade com a conseqüente diminuição dos teores de areia e silte. As substâncias húmicas extraídas de amostras de solos coletados na Bacia do Médio Rio Negro-AM foram caracterizados por análise elementar, ressonância paramagnética eletrônica e ressonância magnética nuclear. Os resultados mostraram que, em função da profundidade, os valores das razões atômicas C/N foram praticamente constantes ao contrário das razões C/O e C/H, as quais diminuíram. As concentrações de radicais livres do tipo semi-quinona das substâncias húmicas variaram de 0,08 a 5,38 x 1018 spins g-1 de carbono, indicando diferença entre os níveis de humificação das SH extraídas de solos amazônicos. As SH extraídas de solos de regiões alagáveis apresentaram maiores concentrações de spins em relação às SH extraídas de regiões não alagáveis, indicando influência da umidade no processo de humificação. / In this research were characterized humic substances (HS) and soil samples extracted from Rio Negro-AM Medium Basin and capacity of complexation (CC) of Hg(II) by HS and constants of exchange between metallic species originally complexed in the HS by Hg(II) had also been determined. The soil samples were characterized through organic material composition, elemental and granulometry analysis. The results showed decreasing of the organic material composition with the depth. According to the results taken, was characterized an increasing in the composition of clay with the depth, but in the other hand, a reduction in silt and sand composition. The humic substances extracted of soil samples from Rio Negro-AM Medium Basin were characterized by elemental analysis, electronic paramagnetic resonance and nuclear magnetic resonance. The results had shown that, in function of the depth, the atomic ratio values C/N were constants, while the atomic ratio C/O and C/H decresed. The concentrations of free radicals of the humic substances were 0.08 to 5.38 x 1018 spins g-1 of carbon, indicating difference in the humification levels of HS extracted from Amazonian soils. The HS extracted in flooded regions showed bigger spins concentrations comparing to ones extracted in unflooded regions, showing that the humidity influences in the process of humification. The NMR of 13C dates showed the sequence of percentage of kind of carbon: aliphatic > ethers/hydroxyls > carboxyls/esthers/amides > aromatics > phenols @ carbonyls of aldehydes and ketones for HS samples extracted of the differences profiles of the Amazonian soils studied.

Made available in DSpace on 2015-03-26T13:53:11Z (GMT). No. of bitstreams: 1 texto completo.pdf: 2564128 bytes, checksum: 2cbe4c261ce82ad997826064088cf96e (MD5) Previous issue date: 2009-07-16 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The use of leguminous as green manure is a widespread practice among family farms of Zona da Mata of Minas Gerais, contributing to the maintenance of the organic systems. With the addition of organic materials in the soil, the humic substances (HS) formation is a important process that increase the cycling time and contribute for soil C and N stocks maintenance. Beyond of the stocks, the constant addition of vegetal waste provides larger biological activity, increase of loads capable to minimize the loss of nutrients of the soil, it increases the aggregation and it reduces the erosion. The leguminous present different characteristics amongst themselves as the biomass production accumulate of nutrients and chemical characteristics. Those ones influence in the formation of the HS in the soil. Those characteristics can also be influenced by the climate of the place and, consequently, influencing in the formation and stabilization of the HS of the soil. The objective of this stud was quantity the C and N contents and the stocks of soil and on HS and evaluate the impact of the leguminous use as green manure in the quality of the HS under cultivation of organic coffee in two different edaphic conditions. Four leguminous species were cultivated between the coffee lines: Arachis pintoi, Calopogonium mucunoides, Stylozanthes guyanensis, Stizolobium aterrimum and spontaneous species, with prevalence of the Brachiaria plantaginea, during 4 years in family farmers of Araponga-MG (northwest exposure) and Pedra Dourada-MG (south exposure) in the area of the Zona da Mata of Minas Gerais. After 4 years of manure, was collected the soil in two depths (0-5 and 5-20 cm) and analyzed the content of Total Organic Carbon (TOC) and Total Nitrogen (TN) in the soil and on HS. In the farms were collected soil samples in volumetric rings to calculate the soil density. For the qualitative evaluation of the HS, they were extracted and purified for obtaining of the fulvic acids (FA) and humic acids (HA). FA and HA were accomplished elementary analysis (CHNO), UV-visible spectroscopy, Infra Red spectroscopy and termogravimetric of the purified material. The manure with leguminous and spontaneous species used as green manure can maintain the content of TOC, TN and the content of C and N on SH in the soil, maintaining the close values to the native forest. In the handling after 4 years of cultivation, just the humic fraction more labile (fulvic acids fraction - FAF) had effect in the treatments, what wasn t observed in the content of the most recalcitrant fractions (humic acids fraction HAF and humin fraction - HF). The content of C and N in HF were more the other fractions. The edaphic conditions promoted effect in the formation and in the stock of C and total N and of HS, and in the farm under conditions of smaller sunstroke (Pedra Dourada), the soil presented larger stocks of C and N. The effect of the position in the landscape (face exposure) it was superior to the effect of the clay content. The analyses to evaluate the quality, was verified that HA possesses larger weight molecular, larger hidrofobicity and larger condensation, for the presence of larger proportion of aromatic compositions and with larger content of C, checking larger structural stability in relation to FA. The farm under larger light incidence and temperature and smaller humidity, due to face of exposure northwest solar presented in FA, larger relationship O/C, relationship E4/E6, relationship 3300/1700 and value of inferior ITG, favoring the formation of FA with larger mass molar in relation to the found in the farm under smaller light incidence and temperature and larger humidity. In HA, the treatments obtained little effect due to the time of handling of the leguminous that they were for only 4 years. Among the leguminous, in FA, the treatment with Calopogonium mucunoides presented low content of C and high content of O in his composition, corroborating for the mass loss the temperature lowers, characterizing a material of easy mineralization. Already the treatment with Stylozanthes guyanensis provided FA with smaller molecular weight. / O uso de leguminosas como adubo verde é uma prática difundida entre os agricultores familiares da Zona da Mata Mineira, contribuindo para a manutenção dos sistemas orgânicos. Com a adição de matériais orgânicos ao solo, a formação das substancias húmicas é um importante processo, que aumenta o tempo de ciclagem e contribui para a manutenção dos estoques de C e N no solo. Além dos estoques, o aporte constante de resíduos vegetais proporciona maior atividade biológica, aumento de cargas capazes de minimizar a perda de nutrientes do solo, aumenta a agregação e diminui a erosão. As leguminosas apresentam características diferentes entre si como a produção de biomassa, acumulo de nutrientes e características químicas, características essas que influenciam na formação das substancias húmicas no solo. Essas características também podem ser influenciadas pelo clima do local e, consequentemente, influenciando na formação e estabilização das substancia húmicas do solo. Objetivou-se quantificar os teores e estoques de C e N no solo e nas substâncias húmicas e avaliar o impacto do uso de leguminosas como adubo verde na qualidade das substâncias húmicas em solos sob cultivo de café orgânico em duas condições edafoclimáticas distintas. Foram cultivadas nas entrelinhas do café quatro espécies de leguminosas: amendoim forrageiro (Arachis pintoi), calopogônio (Calopogonium mucunoides), estilosantes (Stylozanthes guyanensis), mucuna preta (Stizolobium aterrimum) e espécies espontâneas, com predomínio do capim marmelada (Brachiaria plantaginea), durante 4 anos em propriedades de agricultores familiares de Araponga - MG (face de exposição noroeste) e Pedra Dourada - MG (face de exposição sul) de agricultores familiares na região da Zona da Mata de Minas Gerais. Após 4 anos de manejo, foi realizada a coleta de solo em duas profundidades (0-5 e 5-20 cm) e analisado os teores de C e N total no solo e nas substancias húmicas. Por meio de amostras coletadas em anéis volumétricos para a obtenção da densidade do solo. Para a avaliação qualitativa das substâncias húmicas, foram extraídas e purificadas para obtenção dos ácidos fúlvicos (AF) e húmicos (AH). Foram realizadas análise elementar (CHNO), UV-Visível, Infra Vermelho e Termogravimetria do material purificado. O manejo com leguminosas e espécies espontâneas utilizadas como adubo verde pode manter os teores de COT, NT e os teores de C e N das SHs no solo, mantendo os valores próximos à mata nativa. No manejo após 4 anos de cultivo, apenas a fração húmica mais lábil (Fração ácidos Fúlvicos - FAF) teve efeito nos tratamentos, o que não foi observado nos teores das frações mais recalcitrantes (Fração Ácidos Húmicos - FAH e Fração Humina - FH). Os teores de C e N na FH foram superiores as outras frações. O efeito do uso da adubação verde foi verificado principalmente na superfície, com o aumento dos teores de C e N no solo. As condições edafoclimáticas promoveram efeito na formação e no estoque de C e N totais e das SHs, sendo que na propriedade sob condições de menor insolação (Pedra dourada), o solo apresentou maiores estoques de C e N. Foi verificado que os AH possuem maior peso molecular, maior hidrofobicidade e maior condensação, pela presença de maior proporção de compostos aromáticos e com maior teor de C, conferindo maior estabilidade estrutural em relação aos AF. A propriedade rural sob maior incidência de luz e temperatura e menor umidade, devido a face de exposição solar noroeste apresentou no AF, maior relação O/C, relação E4/E6, relação 3300/1700 e valor de ITG inferior, favorecendo a formação de AF com maior massa molar em relação ao encontrado na propriedade sob menor incidência de luz e temperatura e maior umidade. Nos AH, os tratamentos obtiveram pouco efeito devido ao tempo de manejo das leguminosas que foram por apenas 4 anos. Dentre as leguminosas, nos AF, o tratamento com calopogônio apresentou baixo teor de C e elevado teor de O em sua composição, corroborando para a perda de massa a temperatura baixa, caracterizando um material de fácil mineralização. Já o tratamento com estilosantes proporcionou AF com menor peso molecular.

Made available in DSpace on 2015-03-26T13:53:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-07-26 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The lower NH3 volatilization from urea combined with humicified organic material has been attributed to the formation and adsorption of NH4+ due the high potential acidity and CEC. In this context, an experiment was conducted to evaluate NH3 volatilization from grainy urea or coated with humic acids (HA) and humic substances (HS) synthesized from vegetable charcoal (VC). The synthesis of HA and HS was due to the oxidation from charcoal of eucalyptus with HNO3 4.4 mol L-1 boiled under reflux for 4 hours. The HS were obtained directly by filtration. For the HA and HS achievement, It was solubilized in NaOH 0.5 mol L-1 and the fraction of HA was the one that precipitated until acidification to pH 2.0. It was produced grainy urea and urea coated with 5, 10, 15 and 20 % HA or HS. For the granulation, mixtures of urea (p.a.) and HA or HS were fused in 137 ° C and dripped using a pipette on a metal plate. With the fast cooling of the drops, granules in the form of tablets (4 to 5 mm in diameter) were formed. The commercial grainy urea (1-2 mm) was coated with HA or humic substances, using vegetable oil to promote adhesion. Volatilization of NH3 was evaluated in the laboratory in a closed system with continuous air flow, where 100 mg of N-urea was applied to a 100 cm3 surface of a red-yellow Acrisol (<2 mm). The synthesis income was 327 g kg-1 of HA and 784 g kg-1 of humic substance. The acidity of the carboxylic and phenolic groups was estimated through adjusting multiparameter non-linear potentiometric titration curve and, together, corresponded to the CEC of the HA (4750 mmol c kg-1) and HS (4360 mmol c kg-1), which were 9.5 and 8.7 times higher than the VC. The carboxylic groups were predominant either in the VC as in HA and HS, but the HA had a higher content of carboxylic groups and lower content of phenolic groups than the HS. The presence of carboxylic groups was confirmed by the spectra in the infrared range, showing absorption of stretching and deformation, characteristic of C = O bond with 1703.74 and 1706.88 cm-1 waves to the HA and HS, respectively. The urea fusion did not change the N content, but they ranged on average from 36 to 42 % with the addition of 20 to 5 % HA or HS. The highest volatilization rate occurred between 37 and 41 h after application and this time was little influenced by the proportions of HA and HS. Without the addition of HA and HS, the urea in a tablet shape and the commercial grainy urea provided a total volatilization of 517.4 and 468.8 mg g-1 of N-NH3. The largest reductions in evaporation occur with grainy urea (44.5 %) and coated (28.7 %) with 15 and 20 % HA, respectively. In the grainy urea the proportions of HA and HS provided equivalent reduction in evaporation. / A menor volatilização de NH3 a partir da ureia combinada com material orgânico humificado tem sido atribuída à formação e à adsorção do NH4 + em razão da elevada acidez potencial e CTC. Neste contexto, foi conduzido um experimento para avaliar a volatilização de NH3 a partir da ureia granulada ou revestida com ácidos húmicos (AH) e substâncias húmicas (SH) sintetizadas de carvão vegetal (CV). A síntese dos AH e SH se deu pela oxidação do CV de eucalipto com HNO3 4,4 mol L-1 em ebulição sob refluxo por 4h. As SH foram obtidas diretamente por filtragem. Para obter os AH as SH foram solubilizadas em NaOH 0,5 mol L-1 e a fração AH foi aquela precipitada com a acidificação até pH 2,0. Foi produzida ureia granulada e ureia revestida com 5, 10, 15 e 20 % de AH ou SH. Para a granulação as misturas de ureia (p.a.) com AH ou SH foram fundidas a 137 °C e gotejadas com uma pipeta em uma placa metálica. Com o resfriamento rápido das gotas formaram-se grânulos na forma de pastilhas (4 a 5 mm de diâmetro). A ureia granulada comercial (1 a 2 mm) foi revestida com AH ou SH utilizando-se óleo vegetal para favorecer a aderência. A volatilização de NH3 foi avaliada em laboratório em um sistema fechado com fluxo contínuo de ar, onde 100 mg de N-ureia foi aplicada nas superfície de 100 cm3 de um Argissolo Vermelho- Amarelo (< 2 mm). O rendimento na síntese foi de 327 g kg-1 de AH e 784 g kg-1 de SH. A acidez dos grupos carboxílicos e fenólicos foi estimada por meio do ajuste multiparamétrico não linear da curva de titulação potenciométrica e, em conjunto, corresponderam à CTC dos AH (4.750 mmolc kg-1) e SH (4.360 mmolc kg-1), que foram 9,5 e 8,7 vezes maiores do que a do CV. Os grupos carboxílicos predominaram tanto no CV como nos AH e SH, porém os AH apresentaram maior teor de grupos carboxílicos e menor teor de grupos fenólicos do que as SH. A ocorrência dos grupos carboxílicos foi confirmada pelos espectros na faixa do infravermelho que evidenciaram absorção de estiramento e deformação característica da ligação C=O com 1.703,74 e 1.706,88 cm-1 ondas para os AH e SH, respectivamente. A fusão da ureia não alterou o teor de N, mas estes variaram, em média, de 36 a 42 % com a adição de 20 a 5 % de AH ou SH. A maior taxa de volatilização ocorreu entre 37 e 41 h após a aplicação e este tempo foi pouco influenciado pelas proporções de AH e SH. Sem a adição de AH ou SH a ureia na forma de pastilha e a ureia granulada comercial proporcionaram volatilização total de 517,4 e 468,8 mg g-1 de N-NH3. As maiores reduções na volatilização ocorreram com a ureia granulada (44,5 %) e revestida (28,7 %) com 15 e 20 % de AH, respectivamente. Na ureia granulada as proporções de AH e de SH proporcionaram redução equivalente na volatilização.