Challenges for simultaneous nitrification, denitrification, and phosphorus removal in microbial aggregates: mass transfer limitation and nitrous oxide production

The microbial community composition and activity was investigated in aggregates from a lab-scale bioreactor, in which nitrification, denitrification and phosphorus removal occurred simultaneously. The biomass was highly enriched for polyphosphate accumulating organisms facilitating complete removal of phosphorus from the bulk liquid; however, some inorganic nitrogen still remained at the end of the reactor cycle. This was ascribed to incomplete coupling of nitrification and denitrification causing NO3- accumulation. After 2 h of aeration, denitrification was dependent on the activity of nitrifying bacteria facilitating the formation of anoxic zones in the aggregates; hence, denitrification could not occur without simultaneous nitrification towards the end of the reactor cycle. Nitrous oxide was identified as a product of denitrification, when based on stored PHA as carbon source. This observation is of critical importance to the outlook of applying PHA-driven denitrification in activated sludge processes. (c) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.

Candidatus "Anammoxoglobus propionicus" a new propionate oxidizing species of anaerobic ammonium oxidizing bacteria

The bacteria that mediate the anaerobic oxidation of ammonium (anammox) are detected worldwide in natural and man-made ecosystems, and contribute up to 50% to the loss of inorganic nitrogen in the oceans. Two different anammox species rarely live in a single habitat, suggesting that each species has a defined but yet unknown niche. Here we describe a new anaerobic ammonium oxidizing bacterium with a defined niche: the co-oxidation of propionate and ammonium. The new anammox species was enriched in a laboratory scale bioreactor in the presence of ammonium and propionate. Interestingly, this particular anammox species could out-compete other anammox bacteria and heterotrophic denitrifiers for the oxidation of propionate in the presence of ammonium, nitrite and nitrate. We provisionally named the new species Candidatus "Anammoxoglobus propionicus".

Coral disease dynamics and environmental drivers in the northern Florida Keys and Lee Stocking Island, Bahamas

Coral reefs are experiencing declines worldwide and recently coral diseases have been identified as significant contributors to coral mortality. However, little is known regarding the factors that drive coral disease distributions and dynamics. Current knowledge of the organisms that cause coral diseases is also limited, with pathogens having been identified for only 5 of the 21 described coral diseases. The study presented here describes coral disease dynamics in terms of occurrence, prevalence, spatial distribution, and host species susceptibility from 2002--2004 on reefs of the Northern Florida Keys (NFK) and Lee Stocking Island (LSI) in the Bahamas' Exuma chain. In addition, this research investigated the influence of temperature, sediment, and nutrient availability on coral disease prevalence and severity. Finally, microbial communities associated with a polymicrobial disease, black band, were examined to address spatial and temporal variability. ^ Four scleractinian diseases were observed in repeated surveys conducted during June-August of each year: black band disease (BBD), white plague type 2 (WP), dark spots syndrome (DSS), and yellow band disease-(YBD). Coral disease prevalence was generally low in both the NFK and LSI as compared to epizootic levels reported previously in the NFK and other regions of the Caribbean. Disease prevalence and species susceptibility varied spatially and temporally. Massive framework species, including Siderastrea siderea, Colpophyllia natans, and Montastraea annularis, along with relatively smaller colonies of Meandrina meandrites and Dichocoenia stokesi, were most susceptible to disease. Temperature, sedimentation, and dissolved inorganic nitrogen were positively correlated with BBD infections. Furthermore, experimental nutrient enrichment exacerbated coral tissue loss to BBD both in situ and in vivo. Profiling of BBD microbial communities using length heterogeneity PCR revealed variation over space and time, with significantly distinct bacterial assemblages in the NFK, LSI, and US Virgin Islands. ^ This study contributes to knowledge of the relationship between coral diseases and the environment, and facilitates predictions regarding potential changes in coral reef communities under differing environmental conditions. Additionally, this research provides further understanding of coral disease dynamics at both the host and microbial pathogen levels.^

(Supplement 1) Selected chemical composition of Icelandic river waters and bedrock age

In this study we map the spatial distribution of selected dissolved constituents in Icelandic river waters using GIS methods to study and interpret the connection between river chemistry, bedrock, hydrology, vegetation and aquatic ecology. Five parameters were selected: alkalinity, SiO2, Mo, F and the dissolved inorganic nitrogen and dissolved inorganic phosphorus mole ratio (DIN/DIP). The highest concentrations were found in rivers draining young rocks within the volcanic rift zone and especially those draining active central volcanoes. However, several catchments on the margins of the rift zone also had high values for these parameters, due to geothermal influence or wetlands within their catchment area. The DIN/DIP mole ratio was higher than 16 in rivers draining old rocks, but lowest in rivers within the volcanic rift zone. Thus primary production in the rivers is limited by fixed dissolved nitrogen within the rift zone, but dissolved phosphorus in the old Tertiary catchments. Nitrogen fixation within the rift zone can be enhanced by high dissolved molybdenum concentrations in the vicinity of volcanoes. The river catchments in this study were subdivided into several hydrological categories. Importantly, the variation in the hydrology of the catchments cannot alone explain the variation in dissolved constituents. The presence or absence of central volcanoes, young reactive rocks, geothermal systems and wetlands is important for the chemistry of the river waters. We used too many categories within several of the river catchments to be able to determine a statistically significant connection between the chem¬istry of the river waters and the hydrological categories. More data are needed from rivers draining one single hydrological category. The spatial dissolved constituent distribution clearly revealed the difference between the two extremes, the young rocks of the volcanic rift zone and the old Tertiary terrain.

Seawater carbonate chemistry, nutrient uptake and biological processes of coral Stylophora pistillata during experiments, 2011

The effects of ocean acidification and elevated seawater temperature on coral calcification and photosynthesis have been extensively investigated over the last two decades, whereas they are still unknown on nutrient uptake, despite their importance for coral energetics. We therefore studied the separate and combined impacts of increases in temperature and pCO2 on phosphate, ammonium, and nitrate uptake rates by the scleractinian coral S. pistillata. Three experiments were performed, during 10 days i) at three pHT conditions (8.1, 7.8, and 7.5) and normal temperature (26°C), ii) at three temperature conditions (26°, 29°C, and 33°C) and normal pHT(8.1), and iii) at three pHT conditions (8.1, 7.8, and 7.5) and elevated temperature (33°C). After 10 days of incubation, corals had not bleached, as protein, chlorophyll, and zooxanthellae contents were the same in all treatments. However, photosynthetic rates significantly decreased at 33°C, and were further reduced for the pHT 7.5. The photosynthetic efficiency of PSII was only decreased by elevated temperature. Nutrient uptake rates were not affected by a change in pH alone. Conversely, elevated temperature (33°C) alone induced an increase in phosphate uptake but a severe decrease in nitrate and ammonium uptake rates, even leading to a release of nitrogen into seawater. Combination of high temperature (33°C) and low pHT(7.5) resulted in a significant decrease in phosphate and nitrate uptake rates compared to control corals (26°C, pHT = 8.1). These results indicate that both inorganic nitrogen and phosphorus metabolism may be negatively affected by the cumulative effects of ocean warming and acidification.

Distribution of surface sediment properties along the Portuguese western margin

Organic matter contained in surface sediments from four regions on the western Portuguese shelf, which are influenced by coastal upwelling and fluvial input, was analysed with respect to elemental organic carbon (Corg) and nitrogen (Ntotal) content and isotopic carbon and nitrogen ratios (d13Corg, d15N). Corg/Ntotal weight ratios and d13Corg values are interpreted in terms of terrigenous or marine organic matter sources, supported by CaCO3 content. Organic matter in the shelf sediments is mainly of marine origin, with increasing terrigenous components only close to rivers and estuaries. In the northern shelf region the data indicates significant terrigenous supply by the Douro River. North of the Nazaré Canyon organic matter composition implies a mainly marine origin, with a higher terrestrial influence close to the canyon head. Organic matter composition in the central shelf region, which is dominated by the Tagus Estuary and the Tagus prodelta, reveals a change from a continental-type signature within the estuary to a more marine-type signature further to the west and south of the estuary mouth. In the southern region near Cape Sines the geochemical properties clearly reflect the marine origin of sedimentary organic matter. Sedimentary d15N values are interpreted to reflect various degrees of assimilation of seasonally upwelled nitrate, in relation to the upwelling centres. In the estuarine environment, inputs of agriculturally influenced dissolved inorganic nitrogen are reflected in the sediments. No evidence for N2-fixation or denitrification is found. On the central shelf north of the Nazaré canyon, sedimentary d15N values are close to marine d15NO3- and thus indicate a complete NO3- assimilation and N-limitation of marine production. Light d15N values in distal sediments off the Douro River mouth and in samples south of C. Sines reflect high NO3- supply and a close proximity to the seasonal upwelling centres. Particularly in sediments form the Sines region, light d15N values in southern samples reflect stronger upwelling further south.

Cape Lookout cold-water coral area, coral ages and environmental parameters for the last glacial cycle

Climatic and oceanographic changes, as occurring at a glacial-interglacial scale, may alter the environmental conditions needed for the development of prolific cold-water coral reefs and mounds. Studies constraining the temporal distribution of cold-water corals in the NE Atlantic suggested the cyclic changes of the Atlantic Meridional Overturning Circulation as the main driver for the development and dispersal of cold-water coral ecosystems. However, conclusions were hindered by lack of data from the NW Atlantic. Aiming to overcome this lack of data, the temporal occurrence of cold-water corals in the Cape Lookout area along the southeastern US margin was explored by U-series dating. Furthermore, the local influence of the regional water masses, namely the Gulf Stream, on cold-water coral proliferation and occurrence since the Last Glacial Maximum was examined. Results suggest that the occurrence of cold-water corals in the Cape Lookout area is restricted to interglacial periods, with corals being present during the last ~7 kyr and also during the Eemian (~125 ka). The reconstructed local environmental conditions suggest an offshore displacement of the Gulf Stream and increased influence from the Mid-Atlantic Bight shelf waters during the last glacial period. During the deglacial sea level rise, the Gulf Stream moved coastward providing present-day-like conditions to the surface waters. Nevertheless, present-day conditions at the ocean sea floor were not established before 7.5 cal ka BP once the ultimate demise of the Laurentide ice-sheet caused the final sea level rise and the displacement of the Gulf Stream to its present location. Occasional presence of the Gulf Stream over the site during the Mid- to Late Holocene coincides with enhanced bottom current strength and a slightly higher bottom water temperature, which are environmental conditions that are favorable for cold-water coral growth.

Dinâmica de nitrogênio em cultivo heterotrófico a partir de cianobactéria sob o escopo de biorrefinarias agroindustriais

O trabalho teve por objetivo avaliar a dinâmica de nitrogênio, em cultivo heterotrófico, a partir da cianobactéria Aphanothece microscopica Nägeli, sob o escopo de uma biorrefinaria. Neste sentido, foi avaliada a contribuição dos compostos nitrogenados não proteicos, na dinâmica de distribuição do nitrogênio, na biomassa gerada pelo micro-organismo em estudo, quando cultivado em sistema autotrófico e heterotrófico. Para o cultivo em condições autotróficas, foi utilizado o meio padrão BG-11, enquanto que, para o cultivo em condições heterotróficas, foi empregado o efluente da indústria de laticínios. Inicialmente, foi avaliada a contribuição dos pigmentos na fração nitrogenada não proteica tendo como base dois experimentos. No primeiro experimento foi selecionada a melhor condição para a produção de pigmentos, expressos pela clorofila-a em sistema heterotrófico, tendo como base os parâmetros C/N (20, 40 e 60), N/P (5, 10 e 15) e concentração de inóculo (100, 200 e 300 mg.L-1), mediante um planejamento fatorial 23 . Os experimentos foram conduzidos em biorreator heterotrófico a 20°C, pH 7,6 e aeração contínua de 1VVM. A melhor condição de produção de pigmento foi indicada como sendo a 200 mg.L-1 de concentração celular, razões C/N 20 e N/P 10. Com base nestes resultados, um segundo experimento foi delineado, visando avaliar a contribuição de pigmentos na fração de nitrogênio não proteico, bem como avaliar a produção de clorofila-a e ficobiliproteínas (ficocianina, aloficocianina e ficoeritrina), sob influência da luz e do meio de cultivo. Foi possível destacar teores superiores de ficobiliproteínas na biomassa gerada no cultivo heterotrófico. No entanto, com notada diferença (p≤0,05) nos teores de clorofila-a, quando são comparadas as concentrações na biomassa de meios autotróficos (10,7 mg.g-1) e heterotróficos (1,0 mg.g-1). Fato este compensado pelo menor tempo de cultivo registrado para atingir o final do experimento, quando o micro-organismo é cultivado em condições heterotróficas. Fica demonstrado assim, ainda, a importante contribuição dos pigmentos na fração de nitrogênio não proteico. Na sequência, um terceiro e quarto experimentos foram delineados, visando avaliar a influência do nitrogênio inorgânico intracelular na fração não proteica e na produção de proteína, assim como a caracterização da fração proteica quanto ao seu perfil aminoacídico. O estudo da dinâmica do nitrogênio intracelular demonstrou que o N-NH4 + foi a forma nitrogenada predominante, perfazendo importante fração de N-NP, sendo, portanto, os teores de N-NP significativamente dependente dos teores de pigmentos e nitrogênio intracelular. Os aminogramas das biomassas geradas pelos cultivos autotróficos e heterotróficos indicaram como aminoácidos majoritários o ácido glutâmico e aspártico, seguidos por valina, leucina e isoleucina, e como minoritários, lisina, glicina e metionina. O perfil aminoacídico caracterizou-se por apresentar aminoácidos essenciais como isoleucina, metionina + cisteína, fenilalanina + tirosina, valina e treonina em concentrações superiores ao preconizado pela FAO/WHO. A caracterização da fração proteica quanto ao perfil aminoacídico qualificou esta biomassa como fonte potencial de proteína. Os resultados obtidos neste trabalho demonstram a influência e dinâmica de distribuição dos compostos nitrogenados em Aphanothece microscopica Nägeli. Fica demonstrado, ainda, que a implementação do conceito de biorrefino, no tipo de agroindústria estudado, poderá representar importantes possibilidades de aproveitamento sustentável do efluente gerado.

Isolamento, seleção e cultivo de leveduras com potencia biotecnológico para a produção de endo-xilaneses

Xilanases são enzimas que catalisam a hidrólise das xilanas e têm sido em grande parte, obtidas a partir de bolores e bactérias. No entanto poucos estudos têm sido relatados sobre a produção destas enzimas por leveduras. O presente trabalho teve como objetivo isolar leveduras de diferentes fontes vegetais visando à produção de xilanases, além de maximizar sua produção, estudar o uso de diferentes fontes de nitrogênio e cultivar as leveduras em meios contendo coprodutos agroindustriais. As amostras de alimentos e resíduos foram enriquecidas em caldo extrato de malte e levedura e isoladas em Ágar Nutriente Wallerstein, as leveduras isoladas foram, a seguir, avaliadas quanto à capacidade de degradar xilana presente no meio e produzir halos de hidrólise, os quais foram visualizados através do uso do corante vermelho congo. Os micro-organismos selecionados como potenciais produtores de xilanase foram crescidos em meio complexo líquido e as atividades enzimáticas de endoxilanase, β-xilosidase, carboximetilcelulase, celulase total, pH e concentração de biomassa foram avaliadas ao longo de 96 h de cultivo. Dentre as leveduras isoladas, sete foram selecionadas, e a 18Y foi a que apresentou a maior atividade de endo- xilanase (2,7 U.mL-1 ), sendo esta isolada de chicória e identificada como Cryptococcus laurentii. Esta estirpe apresentou capacidade de produzir xilanase com baixos níveis de celulase, sendo assim selecionada neste trabalho. A maximização de endo-xilanase foi avaliada fazendo uso de planejamento experimental onde primeiramente foi realizado um planejamento fracionário 2 6-2 para verificar os efeitos do pH inicial e as concentrações de xilana, peptona, (NH4)2SO4, extrato de levedura e KH2PO4 sobre a atividade enzimática. Após selecionar as variáveis xilana, peptona, pH e extrato de levedura foi realizado um delineamento composto central rotacional (24 ) onde todos os cultivos foram mantidos a 30°C, 150 rpm durante 96 h sendo retiradas alíquotas para determinação das atividades, pH e biomassa. A produção máxima foi de 6,9 U.mL-1 usando 10,0 g.L-1 de extrato de levedura, 10,0 g.L-1 de peptona, 10,0 g.L-1 de xilana, 1,0 g.L-1 de (NH4)2SO4 em pH 6,5 o que permitiu um incremento de mais de 250% sobre a atividade. Posteriormente foram realizados ensaios avaliando diferentes fontes e concentrações de nitrogênio orgânico e inorgânico. A presença de NH4NO3 e (NH₄)₂SO₄ usados na concentração de 3% proporcionaram as maiores atividades de endo-xilanase (6,2 e 6,0 U.mL-1 respectivamente). O sulfato de amônio foi selecionado e fixado em 1 g.L-1 e logo após um planejamento completo 22 foi realizado onde as variáveis xilana e extrato de levedura foram estudadas e as demais fixadas. As condições ótimas estabelecidas para a produção da enzima foram: concentração de xilana de 18,6 g.L-1 , concentração de extrato de levedura de 10 g.L-1 atingindo 14 U.mL-1 . Após a maximização enzimática estudou-se o crescimento de Pichia pastoris NRRL Y-1603 e Cryptococcus laurentti em cinco substratos agroindustriais visando a possibilidade estes substratos substituírem a xilana em cultivos para a produção de endo-xilanase. Os ensaios foram realizados utilizando os subtratos pré-tratados com NaOH 4% e não tratados. Para inserção dos mesmos aos meios de cultivo, estes foram moídos e adicionados na concentração de 2%. O pré-tratamento para todos as fontes de hemicelulose foi eficiente e promoveu aumento nas atividades produzidas. Cryptococcus laurentti apresentou maior atividade enzimática (8,7 U.mL-1 ) em farelo de arroz desengordurado e pré- tratado enquanto que a levedura Pichia pastoris NRRL Y-1603 apresentou sua melhor condição para produção de endo-xilanase quando cultivada em meio contendo casca de aveia e o farelo de arroz pré-tratados, alcançando atividades máximas de 7,6 e 7,5 U.mL-1 .

Impacts of high-nitrate freshwater inputs on macrotidal ecosystems. I. Seasonal evolution of nutrient limitation for the diatom-dominated phytoplankton of the Bay of Brest (France)

The chemical factors (inorganic nitrogen, phosphate, silicic acid) that potentially or actually control primary production were determined for the Bay of Brest, France, a macrotidal ecosystem submitted to high-nitrate-loaded freshwater inputs (winter nitrate freshwater concentrations >700 mu M, Si:N molar ratio as low as 0.2, i.e. among the lowest ever published). Intensive data collection and observations were carried out from February 1993 to March 1994 to determine the variations of physical [salinity, temperature, photosynthetically active radiation (PAR), freshwater discharges] and chemical (oxygen and nutrients) parameters and their impacts on the phytoplankton cycle (fluorescence, pigments, primary production). With insufficient PAR the winter stocks of nutrients were almost nonutilized and the nitrate excess was exported to the adjacent ocean, due to rapid tidal exchange. By early April, a diatom-dominated spring bloom developed (chlorophyll a maximum = 7.7 mu g l(-1); primary production maximum = 2.34 g C m(-2) d(-1)) under high initial nutrient concentrations. Silicic acid was rapidly exhausted over the whole water column; it is inferred to be the primary limiting factor responsible for the collapse of the spring bloom by mid-May. Successive phytoplankton developments characterized the period of secondary blooms during summer and fall (successive surface chlorophyll a maxima = 3.5, 1.6, 1.8 and 1.0 mu g l(-1); primary production = 1.24, 1.18 and 0.35 g C m(-2) d(-1)). Those secondary blooms developed under lower nutrient concentrations, mostly originating from nutrient recycling. Until August, Si and P most likely limited primary production, whereas the last stage of the productive period in September seemed to be N limited instead, this being a period of total nitrate depletion in almost the whole water column. Si limitation of spring blooms has become a common feature in coastal ecosystems that receive freshwater inputs with Si:N molar ratios <1. The peculiarity of Si Limitation in the Bay of Brest is its extension through the summer period.

Scedo-Select III: a new semi-selective culture medium for detection of the Scedosporium apiospermum species complex

International audience

Impact of maintenance of Macrobrachium rosenbergii de Man, 1879 (Crustacea, Decapoda, Palaemonidae) broodstock on the water used in culture ponds

A produção aquícola gera benefícios sociais e econômicos, no entanto, também pode proporcionar impactos ambientais. Os objetivos deste trabalho foram: a) caracterizar os impactos causados pela manutenção de reprodutores do camarão-da-malásia (Macrobrachium rosenbergii) nas características físicas e químicas da água utilizada nos viveiros de cultivo; e b) avaliar a relação entre a biomassa de camarões e o impacto do cultivo na água utilizada no viveiro. Entre janeiro e dezembro de 2004, foram determinadas, mensalmente, a biomassa de M. rosenbergii, por meio de biometria, e as variáveis físicas e químicas da água de abastecimento e do efluente de um viveiro utilizado para a manutenção de reprodutores. Os resultados mostraram que o efluente possui maiores valores de clorofila a, material particulado em suspensão (MPS), pH, oxigênio dissolvido, nitrogênio Kjeldahl total (NKT) e nitrogênio Kjeldahl dissolvido (NKD), nitrogênio inorgânico (NI), fósforo (PT) e fósforo dissolvido (PD) e P-ortofosfato do que a água de abastecimento do viveiro. A maior biomassa de M. rosenbergii ocorreu em abril (127,0 g.m-2) e a menor em agosto (71,5 g.m-2) e houve correlações lineares positivas entre a biomassa de camarões e a intensidade do aumento de NKT, NKD, NI, PT e PD da água utilizada no viveiro. A manutenção de reprodutores de M. rosenbergii aumentou a clorofila-a, MPS, nitrogênio e fósforo da água utilizada no viveiro. Além disso, o aumento da biomassa de camarões intensifica a exportação de nitrogênio e fósforo do viveiro pelo efluente.