Submitted by Marco Antônio de Ramos Chagas (mchagas@ufv.br) on 2017-07-20T13:35:31Z No. of bitstreams: 1 texto completo.PDF: 286988 bytes, checksum: d9f6fc5b932ecff9285c6e82130b932f (MD5) / Made available in DSpace on 2017-07-20T13:35:31Z (GMT). No. of bitstreams: 1 texto completo.PDF: 286988 bytes, checksum: d9f6fc5b932ecff9285c6e82130b932f (MD5) Previous issue date: 2001-03-06 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Este trabalho teve como objetivo analisar alguns aspectos da tolerância ao Al em dois cultivares de arroz: Maravilha (sensível: CNA-6843-1) e Fernandes (tolerante: CNA-1158), em solução nutritiva. Num primeiro experimento, plantas dos dois cultivares foram expostas ao Al em soluções nutritivas com diferentes proporções de NO 3- :NH 4+ e avaliados os efeitos do Al sobre o crescimento das plantas e sobre a liberação de H + para a solução nutritiva. Independente da proporção NO 3- :NH 4+ , tanto o comprimento quanto a produção de matéria seca de raízes e parte aérea diminuíram na presença de Al, nos dois cultivares, principalmente no sensível. O cultivar tolerante mostrou-se mais eficiente no consumo de H + quando o N disponível estava exclusivamente na forma nítrica. O sensível, entretanto, mostrou-se mais eficiente na liberação de H + para a solução nutritiva quando o N foi fornecido sob duas formas ou exclusivamente na forma amoniacal. Num segundo experimento, as plantas dos dois cultivares foram expostas a duas concentrações de Al e avaliados seus efeitos sobre os teores, as taxas de absorção líquida e os índices de eficiência de utilização de alguns elementos minerais, nas duas partes das plantas. Na presença de Al as plantas apresentaram teores mais elevados deste elemento nas duas partes da planta, principalmente nas raízes. O cultivar tolerante mostrou-se mais eficiente na retenção deste elemento nas raízes. Na presença de Al, observou-se redução nos teores de todos os elementos minerais estudados, tanto nas raízes como na parte aérea, exceto no de Ca nas raízes e no de K na parte aérea, do cultivar tolerante. De modo geral, as reduções nos teores destes elementos minerais foram mais intensas no cultivar sensível, nas duas partes das plantas. As taxas de absorção líquida, sofreram reduções crescentes com os níveis de Al na solução nutritiva, tendo o cultivar tolerante apresentado, de modo geral, taxas mais elevadas para todos os elementos minerais estudados. O efeito do Al sobre os índices de eficiência de utilização dos elementos minerais estudados foi variado. No cultivar tolerante, observou-se aumento nos índices de eficiência de utilização de P, Ca e Mg, permanecendo inalterado o de K. No cultivar sensível, os índices de eficiência de utilização de P e K diminuíram, o de Ca permaneceu inalterado e o de Mg aumentou. Na ausência de Al os cultivares não diferiram entre si quanto a estes índices, mas na presença deste elemento, seus valores foram, de modo geral, mais elevados no cultivar tolerante. / The objective of this work was to study some aspects of Al tolerance in two rice cultivars: Maravilha (sensitive: CNA-6843-1) and Fernandes (tolerant: CNA:1158), grown in nutrient solution. In a first experiment, plants of the two cultivars were exposed to Al in nutrient solutions with different NO 3- :NH 4+ proportions and the Al effects on plant growth and on H + exsudation to the nutrient solution were evaluated. Independent of the NO 3- :NH 4+ proportion, both the length and dry matter yield of roots and shoots decreased in Al presence, in both cultivars, mainly in the sensitive. The tolerant cultivar showed higher efficiency in the H + consumption in nutrient solution with the nitric form. The sensitive, however, showed higher the N exclusively in efficiency in the exudation of H + to the nutrient solution, when N was supplied in both N forms or exclusively in the ammonium form. In a second experiment, plants of both cultivars were exposed to two Al concentrations and its effects on the contents, on the net absorption rates and on the use efficiency indices of some mineral elements, were evaluated in both parts of the plants. In Al presence the plants exhibited increased contents of these element in both parts of the plant, mainly in the roots. The tolerant cultivar was more efficient in the Al retention in the roots than the sensitive one. In Al presence, the contents of all mineral elements decreased in both roots and shoots, except the content of Ca in the roots and of K in the shoots, of the tolerant cultivar. In general, the reductions in the mineral contents were stronger in the sensitive cultivar, in both parts of the plants. The net absorption rates reductions increased with Al levels in the nutrient solution. The tolerant cultivar, in general, exhibited higher absorption rates of all mineral elements studied. Aluminum effect on the use efficiency indices was quite variable. In the tolerant cultivar, the use efficiency indices of P, Ca and Mg increased and of K remained unaffected. In the sensitive cultivar, the use efficiency indices of P and K decreased, of Ca remained unaffected and of Mg increased. In Al absence there was no difference between cultivars with respect to these indices, but in the presence of this element, the tolerant cultivar showed higher values. / Dissertação importada do Alexandria

No description available.

Atmospheric deposition of nitrogen has been implicated in the destruction of seagrass beds and in the decline of water quality of Tampa Bay, Florida. The objective of this research was to determine the tendency for air masses of different origins to wet-deposit nitrate and ammonium species to the bay. Precipitation chemistry data was obtained via the NADP AIRMoN Gandy Bridge monitoring site for the period of 1 August 1996 through 31 December 2000. Rainfall events were classified by using the NOAA HYSPLIT trajectory model, precipitation chemistry data, and tropical storm history data. Average nitrate and ammonium concentrations and nitrogen fluxes were calculated based upon the chosen categories. The average annual nitrogen flux for nitrate and ammonium were 2.1 kg/ha/yr and 1.4 kg/ha/yr, respectively. For trajectory-classified data, the lowest nitrate and ammonium nitrogen fluxes were observed with air masses from the west and south, over the Gulf of Mexico. The highest ammonium nitrogen flux was seen from trajectories from the east, while local trajectories demonstrated the highest average nitrate nitrogen flux. For chemically-classified data, the highest nitrate and ammonium fluxes were associated with the local combustion classification. Rainfall from tropical weather systems deposited lower average nitrate nitrogen fluxes than non-tropical events, but ammonium nitrogen fluxes were the same between tropical and non-tropical precipitation. Even the events representing the cleanest air masses contributing precipitation to Tampa Bay had nitrate and ammonium concentrations more than two times the background concentrations associated with the northern hemisphere.

The quality control laboratory at PolyPeptide has been performing analysis based on ion chromatography (IC) of peptides. A method for separation of negative ions (acetate, chloride etc) is already in use. Lately there has been a need for developing a method to separate positive ions, like ammonium ions, in peptides. The task of this project was to develop a method for ion chromatography, suitable for determination of ammonium ions in the substance D-alanine amide, hydrochloride (D-Ala-NH2∙HCl). The D-alanine amide, hydrochloride is an amino acid derivative used as starting material for chemical synthesis of peptides. These peptides are used as active pharmaceutical substances. It is most possible to find ammonium chloride in amino acids substances which content amid group, as D-Ala-NH2∙HCl. The ammonium chloride is formed during the synthesis of these amino acid derivatives. The reason for analysing ammonium ions is that there is some risk to find undesired impurities in the product. These impurities are difficult to remove.

La composition isotopique (¹³C/¹²C) du CO₂ respiré par les feuilles et les racines dans l'obscurité et celle des substrats respiratoires putatifs, y compris les sucres solubles et les acides organiques (malate et citrate), l'activité de la PEPc ainsi que les échanges gazeux des feuilles ont été déterminés sur le haricot (Phaseolus vulgaris L.) et l’épinards (Spinacia oleracea L.) cultivés dans du sable avec différents rapports NH₄⁺: NO₃⁻ dans N fourni. Le CO₂ respiré par les feuilles était enrichi en ¹³C sous NO₃⁻ s’appauvrissant progressivement avec l’augmentation de la fraction de NH₄⁺ dans le N fourni, tandis que la composition isotopique du CO₂ issu de la respiration des racines restait inchangée quel que soit le rapport NH₄⁺: NO₃⁻. Nous avons suggéré qu'une plus grande quantité de pools enrichis en ¹³C fixés par la PEPc via la voie anaplérotique contribuait à la respiration foliaire sous NO₃⁻. Cependant, un effet similaire dans les racines attendu sous NH₄⁺ a été masqué en raison d'une refixation (par la PEPc) du CO₂ respiré (appauvrie en ¹³C). De manière inattendue, les modifications de la composition isotopique du C des métabolites individuels, leurs quantités, ainsi que l'activité de l’enzyme PEPc, présentaient des profils différents entre les deux espèces étudiées. Des expériences de double marquage (¹³C et ¹⁵N) sont nécessaires pour mieux comprendre l’impact de la plasticité métabolique du TCA sur l’écart isotopique entre le malate et le citrate et sur la composition isotopique du CO₂ respiré chez différentes espèces sous différentes formes d’azote. / C-isotope composition of leaf- and root-respired CO₂ in the dark and that of putative respiratory substrates including soluble sugars and organic acids (malate and citrate), PEPc activity, as well as leaf gas exchanges were determined on bean (Phaseolus vulgaris L.) and spinach (Spinacia oleracea L.) plants grown in sand with varying ratios of NH₄⁺: NO₃⁻ in supplied N. Leaf-respired CO₂ was ¹³C enriched under NO₃⁻ nutrition and became progressively ¹³C depleted with increasing amount of NH₄⁺ in supplied N, while C-isotope composition of root-respired CO₂ remained unchanged across N-type gradient. We suggested that a higher amount of ¹³C enriched C-pools fixed by PEPc through anaplerotic pathway contributed to respired CO₂ in leaves under NO₃⁻ nutrition. However, a similar effect in roots expected under NH4+ nutrition was masked because of a rather ¹³C depleted C source (respired CO₂) refixation by PEPc. Unexpectedly, the changes in C-isotope composition of individual metabolites and their amounts as well as PEPc activity exhibited different patterns between the two species. Double labelling experiments (¹³C and ¹⁵N) are needed for better understanding the impact of metabolic plasticity of TCA on isotopic gap between malate and citrate and on C-isotope composition of respired CO₂ in different species under varying N-type nutrition.

The constructed wetlands, which have been being created in the Czech Republic since early 90s, are designed so they do not allow successful removal of ammonia nitrogen. Almost all of these 300 registered constructed wetlands suffer from the colmatage which leads to the lowest effectiveness of treatment in nearly all parameters. The main effort of the Institute of landscape water management is to get rid of bad reputation and to clarify the reason that leads to unsatisfactory conditions of constructed wetlands. The diploma thesis focuses on the selected constructed wetland in Dražovice (850 EO), which removes ammonia nitrogen with the effect just below the limit values in the long term. The pressure of the Morava river basin and Department of environment (Building Authority in Bučovice) has led to the necessity of taking measures leading to the increase of treatment efficiency especially ammonia nitrogen. The diploma thesis includes both, actual project documentation concentrating on the adjustment of watercourse to nitrify ammonia nitrogen, and effective choice of suitable filtration material supported by testing method in situ. The practical part of the thesis deals with the implementation of the technical solution according to the project documentation. The adjustment of the constructed wetland is now nearly before completion. Last adjustments will be taking place during the spring 2017, the final inspection as well as the evaluation of the reconstruction will take place at the end of this year.

A (Na+,K+)-ATPase presente no tecido branquial dos crustáceos osmorreguladores é um componente essencial de seu sistema de regulação iônica e osmótica. Esta enzima também apresenta um papel relevante no processo de excreção ativa de NH4+ através do tecido branquial dos crustáceos. Uma fração microsomal rica em (Na+, K+)ATPase foi preparada por centrifugação diferencial a partir de um homogeneizado do tecido branquial de Callinectes ornatus. A centrifugação em gradiente de sacarose revelou a presença de um unico pico de proteina com atividade ATPase, mas a eletroforese em gel de poliacrilamida em condições desnaturantes revelou a presença de várias bandas protéicas. O uso do anticorpo monoclonal 5 contra a subunidade da (Na+, K+) ATPase, revelou a presença de uma única banda proteica de 110 kDa com atividade (Na+, K+)?ATPase. A (Na+, K+) ATPase hidrolisou o PNPP (V= 52,0 ± 2,0 U/mg e K0,5 = 1,1 ± 0,1 mM) através de interações sítio-sítio (nH= 1,6). A modulação da enzima pelos íons magnésio (V= 52,3 ± 1,3 U/mg e K0,5 = 1,1 ± 0,05 mM), potássio (V= 51,4 ± 1,5 U/mg e K0,5 = 2,3 ± 0,1 mM) e amônio (V= 56,7 ± 2,6 U/mg e K0,5 = 9,8 ± 0,4 mM) ocorreu através de interações sítio-sítio. Os íons sódio atuaram como inibidores da atividade K+-fosfatase da enzima (Ki= 1,7 ± 0,1 mM) e a ouabaína inibiu cerca de 80% a atividade PNPPase independentemente da presença de íons amônio. A (Na+, K+) ATPase hidrolisou o ATP de acordo com cinética de Michelis-Menten, com KM= 0,16 0,01 mM e V= 116,3 5,6 U/mg, enquanto a modulação da atividade da enzima pelos íons magnésio (V= 111,0 ± 5,4 U/mg e K0,5= 0,54 ± 0,03 mM), sódio (V= 110,6 ± 5,3 U/mg e K0,5= 6,3 ± 0,3 mM), potássio (V= 116,0 ± 5,5 U/mg e K0,5= 1,5 ± 0,1 mM) e amônio (V= 173,3 ± 5,4 U/mg e K0,5= 5,4 ± 0,3 mM) ocorreram através de interações sítio-sítio. Também foi observado que na presença de concentrações crescentes de ions amonio, a estimulação da atividade (Na+,K+)-ATPase pelo íons potássio acarretou um aumento de 50% na atividade específica da enzima. A ouabaína inibiu cerca de 86% a atividade (Na+,K+)-ATPase com Ki= 74,5 ?M, sugerindo a presença de 14% de fosfatases e/ou outras ATPase contaminantes. Este é o primeiro trabalho onde se observa uma estimulação sinergística da atividade K-fosfatase da (Na+,K+)-ATPase de crustáceo pelos íons potássio e amônio. Os resultados cinéticos obtidos para a (Na+,K+)-ATPase branquial de Callinectes ornatus, analisados em conjunto com os já descritos para outras espécies de crustáceos poderão abrir novas perspectivas em relação ao papel dessa enzima na adaptação fisiológica-bioquímica, bem como para a sobrevivência desses animais em diferentes ambientes. / (Na+, K+)-ATPase present on branchial tissue osmoregulatory crustaceans is an essential component of their osmotic and ionic regulation system. Apparently, this enzyme also have a relevant role in the active excretion de NH4+ through the branchial crustacean tissue. A (Na+, K+) ATPase-rich microsomal fraction was prepared by differential centrifugation from Callinectes ornatus homogenized branchial tissue. The sucrose gradient sucrose centrifugation showed the presence of a single protein peak with ATPase activity, and SDS-PAGE revealed the presence of several proteins bands. The use of the 5 monoclonal antibody, against the ? subunit, revealed the presence of a unique protein band of 110 kDa corresponding to the (Na+, K+) ATPase. (Na+, K+) ATPase hydrolyzed the PNPP (V= 52.0 2.0 U/mg and K0.5= 1.1 0.1 mM) through the site-site interactions (nH= 1.6). The modulation of (Na+, K+) ATPase by magnesium (V= 52.3 1.3 U/mg and K0.5= 1.1 ? 0.05 mM), potassium (V= 51.4 1.5 U/mg and K0.5= 2.3 0.1 mM) and ammonia ions (V= 56.7 2.7 U/mg and K0.5= 9.8 ? 0.4 mM) followed cooperative kinetics. However, sodium ions inhibited PNPPase activity of (Na+, K+)?ATPase with Ki= 1.7 0.1 mM. Ouabain also inhibited up to 80% the total activity PNPPase independent of the presence of ammonium ions. The hydrolysis of ATP by (Na+, K+) ATPase followed Michaelis-Menten kinetics with Km= 0.16 0.01 mM and V= 116.3 5.6 U/mg, while enzyme modulation by magnesium (V= 111.0 5.4 U/mg and K0.5= 0.54 0.03 mM), sodium (V= 110.6 5.3 U/mg and K0.5= 6.3 0.3 mM), potassium (V= 116.0 5.5 U/mg and K0.5= 1.5 ? 0.1 mM) and ammonium ions (V= 173.3 . Interestingly, the stimulation of (Na+, K+)-ATPase activity by potassium ions in the presence of increasing concentration of ammonium ions to K+ resulted in a 50% higher specific activity. Ouabain inhibited approximately 86% the activity (Na+, K+) ATPase with (Ki= 74.5 M), suggesting the presence of about 14% of phosphatases and/or other ATPases. This is the first work showing synergistic stimulation of crustacean (Na+, K+) ATPase by potassium and ammonium ions when PNPP is used a substrate. The results reported herein for Callinectes ornatus branchial (Na+, K+) ATPase might open new perspectives concerning the physiological adaption and the survival of these animals in different environmental.

L'ammoniac atmosphérique (NH3) est principalement émis par l'agriculture. C'est le principal composé alcalin de l'atmosphère et il joue un rôle essentiel dans la chimie de l'atmosphère. Les dépôts humides et secs d'NHx (NH3 + NH4+) ont des effets négatifs directs (toxicité à fortes doses) ou indirects acidification, eutrophisation, baisse de la résistance aux stress) sur les écosystèmes naturels ou cultivés.<br />La plante peut être un puits ou une source d'NH3 pour l'atmosphère. Le sens et l'intensité des échanges dépendant du gradient de concentration entre la cavité sous stomatique et l'air. La concentration en ammoniac dans la cavité sous stomatique appelée aussi point de compensation stomatique pour l'ammoniac est déterminée par la température de la feuille, la concentration en ammoniaque ([NH4+]ap) et le pH du liquide apoplastique. Ces deux derniers paramètres dépendent du métabolisme azoté de la feuille et des diverses entrées et sorties de composés à travers le xylème et le phloème. Jusqu'à présent, seuls des modèles empiriques ont été conçus pour modéliser le point de compensation stomatique pour l'ammoniac.<br />Pour analyser le determinisme du point de compensation pour l'ammoniac, nous avons conçu une expérimentation pour mesurer sa variabilité pour de jeunes plantes de colza en fonction de la nutrition azotée et des conditions jour et nuit. Deux méthodes de mesures ont été utilisées : l'extraction d'apoplaste et la chambre à flux. Les deux méthodes montrent une corrélation positive entre la concentration en ammonium dans la solution nutritive et le point de compensation. Par contre la concentration en nitrate a peu d'influence sur le point de compensation de même qu'une alimentation mixte ammonium et nitrate réduit le point de compensation par rapport a une alimentation a même concentration en ammonium. Le point de compensation mesuré de jour n'est pas significativement différent du point de compensation mesuré de nuit même si il a tendance à être plus fort.<br />Le modèle construit est un modèle a compartiment ; il est couplé à un modèle de continuum sol plante atmosphère donnant des flux d'eau et en particulier la conductance stomatique. Il intègre un modèle de photosynthèse et les principales sources et puits d'ammoniac dans une cellule. Le modèle montre une forte dépendance du point de compensation stomatique du flux d'eau, de la température et de la concentration en ammonium dans le xylème.

The role of sodium chloride (NaCl) particles and ammonium sulfate ((NH4)2SO4) particles on the initial atmospheric corrosion of copper and zinc was investigated under in situ and ex situ conditions using microgravimetry, FTIR spectroscopy, ion chromatography, scanning electron microscopy with x-ray microanalysis and the scanning Kelvin probe. For the first time, in situ infrared spectra were collected on a micron level during particle induced atmospheric corrosion using a recently developed experimental set-up for in situ FTIR microspectroscopy. Lateral distribution of corrosion and reaction products on copper and zinc surfaces was determined and could be connected with the mechanisms of the initial particle induced corrosion. The recently discovered secondary spreading effect from NaCl electrolyte droplets on metal surfaces was studied under in situ conditions and the effect of CO2 on the spreading process was elaborated. The ambient level of CO2 (350 ppm, 1 ppm = 10-6 volume parts) results in a relatively low secondary spreading effect, whereas the lower level of CO2 (&lt;5 ppm) causes a much faster secondary spreading effect over a large area. At low CO2 concentration alkaline conditions will prevail in the cathodic area, leading to large changes in the surface tension at the oxide/electrolyte interface in the peripherical parts of the droplet. This induces a surface tension driven convective flow of electrolyte from the NaCl droplet. The continuous growth of the secondary spreading area at low CO2 concentration is possible due to the galvanic coupling with the droplet leading to transport of sodium ions to this region and maintenance of the alkaline conditions. At 350 ppm CO2, carbonate formation in the secondary spreading area results in lowering of the pH, increasing the surface tension of the oxide/electrolyte interface and inhibiting the secondary spreading. CO2 strongly affects the NaCl-induced atmospheric corrosion rate of copper. The overall influence of CO2 and NaCl depends on at least three identified mechanisms. At low NaCl particle density, CO2 affects the secondary spreading effect from the electrolyte droplet. This leads to a larger effective cathodic area at low CO2 concentration and a higher corrosion rate. The more alkaline surface electrolyte present at low CO2 concentration also affects the formation of corrosion products and the amount of soluble copper chloride. Whereas the presence of larger amounts of soluble chloride tends to increase the corrosion rate, the formation of CuO results in a more protective surface film which decreases the corrosion rate. This effect was observed at higher NaCl particle densities, where the secondary spreading areas overlapped with adjacent NaCl particle clusters. The formation of CuO leads to lower corrosion rates compared to ambient CO2 concentration in which this phase was not formed. For zinc, the formation of a more protective corrosion product layer was not observed and the corrosion rate is generally higher for low than for ambient CO2 concentration. The presence of NaCl particles on the metal surfaces strongly affects the SO2 interaction with the metal surfaces. The oxidation of S(IV) turned out to be fast at the area of the NaCl-containing electrolyte droplet, both for copper and zinc. On copper surfaces, both sulphate (SO4 2-) and dithionate (S2O6 2-) ions formed which is consistent with a copper catalysed reaction route for sulfite oxidation including the formation of a Cu(II)–sulfito complex as an important step. For zinc, a surface mediated sulfite oxidation process leads to rapid formation of sulphate in the electrolyte droplet area. The presence of SO2 strongly inhibits the secondary spreading due to the decrease in pH induced by absorption of SO2 in the cathodic areas. The presence of gaseous oxidants, such as NO2 and O3, has previously been considered as an important prerequisite for the oxidation of sulfite on copper. The results obtained here suggest that the formation of local electrochemical cells induced by deposited NaCl particles could be another important route for S(IV)- oxidation to sulfate formation. On copper, SO2 was also found to promote the formation of less soluble copper chlorides, such as paratacamite (Cu2(OH)3Cl) and nantokite (CuCl). The electrolyte droplet was dried after 24 hours of exposure due to the formation of less soluble paratacamite (Cu2(OH)3Cl) and nantokite (CuCl) and led to a decrease in the corrosion rate. Thus, SO2 alone promotes the corrosion rate of copper, whereas in the presence of NaCl particles the corrosion rate of copper may slow down due to the formation of insoluble copper chloride compounds. The lateral distribution of corrosion products after exposure of NaCl contaminated copper and zinc surfaces to humid air with gaseous pollutants is a result of the formation of local electrochemical cells at the particles and concomitant differences in chemical composition and pH. For (NH4)2SO4 deposited copper and zinc surfaces the corrosion effects increase with the amount of pre-deposited particles and with the exposure time. On copper, the size of the particles affects the corrosion rate, smaller particles resulting in a higher corrosion rate than larger particles at equal amount of deposition. The formation of Cu2O was the dominant corrosion product after exposure longer than 10 days. (NH4)2SO4 particles result in enhanced Cu2O formation on copper due to a reaction sequence involving catalysis by NH3. The corrosion of copper by (NH4)2SO4 particles was much larger than that induced by NaCl particles. However, for zinc, the (NH4)2SO4 particles lead to smaller corrosion effects than those of NaCl particles. For both particles, significant corrosion attack was observed at relative humidity (RH) lower than the deliquescence point of the salts. / QC 20101001

A (Na+,K+)-ATPase presente no tecido branquial dos crustáceos osmorreguladores é um componente essencial de seu sistema de regulação iônica e osmótica. Esta enzima também apresenta um papel relevante no processo de excreção ativa de NH4+ através do tecido branquial dos crustáceos. Uma fração microsomal rica em (Na+, K+)ATPase foi preparada por centrifugação diferencial a partir de um homogeneizado do tecido branquial de Callinectes ornatus. A centrifugação em gradiente de sacarose revelou a presença de um unico pico de proteina com atividade ATPase, mas a eletroforese em gel de poliacrilamida em condições desnaturantes revelou a presença de várias bandas protéicas. O uso do anticorpo monoclonal 5 contra a subunidade da (Na+, K+) ATPase, revelou a presença de uma única banda proteica de 110 kDa com atividade (Na+, K+)?ATPase. A (Na+, K+) ATPase hidrolisou o PNPP (V= 52,0 ± 2,0 U/mg e K0,5 = 1,1 ± 0,1 mM) através de interações sítio-sítio (nH= 1,6). A modulação da enzima pelos íons magnésio (V= 52,3 ± 1,3 U/mg e K0,5 = 1,1 ± 0,05 mM), potássio (V= 51,4 ± 1,5 U/mg e K0,5 = 2,3 ± 0,1 mM) e amônio (V= 56,7 ± 2,6 U/mg e K0,5 = 9,8 ± 0,4 mM) ocorreu através de interações sítio-sítio. Os íons sódio atuaram como inibidores da atividade K+-fosfatase da enzima (Ki= 1,7 ± 0,1 mM) e a ouabaína inibiu cerca de 80% a atividade PNPPase independentemente da presença de íons amônio. A (Na+, K+) ATPase hidrolisou o ATP de acordo com cinética de Michelis-Menten, com KM= 0,16 0,01 mM e V= 116,3 5,6 U/mg, enquanto a modulação da atividade da enzima pelos íons magnésio (V= 111,0 ± 5,4 U/mg e K0,5= 0,54 ± 0,03 mM), sódio (V= 110,6 ± 5,3 U/mg e K0,5= 6,3 ± 0,3 mM), potássio (V= 116,0 ± 5,5 U/mg e K0,5= 1,5 ± 0,1 mM) e amônio (V= 173,3 ± 5,4 U/mg e K0,5= 5,4 ± 0,3 mM) ocorreram através de interações sítio-sítio. Também foi observado que na presença de concentrações crescentes de ions amonio, a estimulação da atividade (Na+,K+)-ATPase pelo íons potássio acarretou um aumento de 50% na atividade específica da enzima. A ouabaína inibiu cerca de 86% a atividade (Na+,K+)-ATPase com Ki= 74,5 ?M, sugerindo a presença de 14% de fosfatases e/ou outras ATPase contaminantes. Este é o primeiro trabalho onde se observa uma estimulação sinergística da atividade K-fosfatase da (Na+,K+)-ATPase de crustáceo pelos íons potássio e amônio. Os resultados cinéticos obtidos para a (Na+,K+)-ATPase branquial de Callinectes ornatus, analisados em conjunto com os já descritos para outras espécies de crustáceos poderão abrir novas perspectivas em relação ao papel dessa enzima na adaptação fisiológica-bioquímica, bem como para a sobrevivência desses animais em diferentes ambientes. / (Na+, K+)-ATPase present on branchial tissue osmoregulatory crustaceans is an essential component of their osmotic and ionic regulation system. Apparently, this enzyme also have a relevant role in the active excretion de NH4+ through the branchial crustacean tissue. A (Na+, K+) ATPase-rich microsomal fraction was prepared by differential centrifugation from Callinectes ornatus homogenized branchial tissue. The sucrose gradient sucrose centrifugation showed the presence of a single protein peak with ATPase activity, and SDS-PAGE revealed the presence of several proteins bands. The use of the 5 monoclonal antibody, against the ? subunit, revealed the presence of a unique protein band of 110 kDa corresponding to the (Na+, K+) ATPase. (Na+, K+) ATPase hydrolyzed the PNPP (V= 52.0 2.0 U/mg and K0.5= 1.1 0.1 mM) through the site-site interactions (nH= 1.6). The modulation of (Na+, K+) ATPase by magnesium (V= 52.3 1.3 U/mg and K0.5= 1.1 ? 0.05 mM), potassium (V= 51.4 1.5 U/mg and K0.5= 2.3 0.1 mM) and ammonia ions (V= 56.7 2.7 U/mg and K0.5= 9.8 ? 0.4 mM) followed cooperative kinetics. However, sodium ions inhibited PNPPase activity of (Na+, K+)?ATPase with Ki= 1.7 0.1 mM. Ouabain also inhibited up to 80% the total activity PNPPase independent of the presence of ammonium ions. The hydrolysis of ATP by (Na+, K+) ATPase followed Michaelis-Menten kinetics with Km= 0.16 0.01 mM and V= 116.3 5.6 U/mg, while enzyme modulation by magnesium (V= 111.0 5.4 U/mg and K0.5= 0.54 0.03 mM), sodium (V= 110.6 5.3 U/mg and K0.5= 6.3 0.3 mM), potassium (V= 116.0 5.5 U/mg and K0.5= 1.5 ? 0.1 mM) and ammonium ions (V= 173.3 . Interestingly, the stimulation of (Na+, K+)-ATPase activity by potassium ions in the presence of increasing concentration of ammonium ions to K+ resulted in a 50% higher specific activity. Ouabain inhibited approximately 86% the activity (Na+, K+) ATPase with (Ki= 74.5 M), suggesting the presence of about 14% of phosphatases and/or other ATPases. This is the first work showing synergistic stimulation of crustacean (Na+, K+) ATPase by potassium and ammonium ions when PNPP is used a substrate. The results reported herein for Callinectes ornatus branchial (Na+, K+) ATPase might open new perspectives concerning the physiological adaption and the survival of these animals in different environmental.