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      Denitrification and Anammox in Tropical Aquaculture Settlement Ponds: An Isotope Tracer Approach for Evaluating N 2 Production

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          Abstract

          Settlement ponds are used to treat aquaculture discharge water by removing nutrients through physical (settling) and biological (microbial transformation) processes. Nutrient removal through settling has been quantified, however, the occurrence of, and potential for microbial nitrogen (N) removal is largely unknown in these systems. Therefore, isotope tracer techniques were used to measure potential rates of denitrification and anaerobic ammonium oxidation (anammox) in the sediment of settlement ponds in tropical aquaculture systems. Dinitrogen gas (N 2) was produced in all ponds, although potential rates were low (0–7.07 nmol N cm −3 h −1) relative to other aquatic systems. Denitrification was the main driver of N 2 production, with anammox only detected in two of the four ponds. No correlations were detected between the measured sediment variables (total organic carbon, total nitrogen, iron, manganese, sulphur and phosphorous) and denitrification or anammox. Furthermore, denitrification was not carbon limited as the addition of particulate organic matter (paired t-Test; P = 0.350, n = 3) or methanol (paired t-Test; P = 0.744, n = 3) did not stimulate production of N 2. A simple mass balance model showed that only 2.5% of added fixed N was removed in the studied settlement ponds through the denitrification and anammox processes. It is recommended that settlement ponds be used in conjunction with additional technologies (i.e. constructed wetlands or biological reactors) to enhance N 2 production and N removal from aquaculture wastewater.

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          Most cited references21

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          Production of N(2) through anaerobic ammonium oxidation coupled to nitrate reduction in marine sediments.

          In the global nitrogen cycle, bacterial denitrification is recognized as the only quantitatively important process that converts fixed nitrogen to atmospheric nitrogen gas, N(2), thereby influencing many aspects of ecosystem function and global biogeochemistry. However, we have found that a process novel to the marine nitrogen cycle, anaerobic oxidation of ammonium coupled to nitrate reduction, contributes substantially to N(2) production in marine sediments. Incubations with (15)N-labeled nitrate or ammonium demonstrated that during this process, N(2) is formed through one-to-one pairing of nitrogen from nitrate and ammonium, which clearly separates the process from denitrification. Nitrite, which accumulated transiently, was likely the oxidant for ammonium, and the process is thus similar to the anammox process known from wastewater bioreactors. Anaerobic ammonium oxidation accounted for 24 and 67% of the total N(2) production at two typical continental shelf sites, whereas it was detectable but insignificant relative to denitrification in a eutrophic coastal bay. However, rates of anaerobic ammonium oxidation were higher in the coastal sediment than at the deepest site and the variability in the relative contribution to N(2) production between sites was related to large differences in rates of denitrification. Thus, the relative importance of anaerobic ammonium oxidation and denitrification in N(2) production appears to be regulated by the availability of their reduced substrates. By shunting nitrogen directly from ammonium to N(2), anaerobic ammonium oxidation promotes the removal of fixed nitrogen in the oceans. The process can explain ammonium deficiencies in anoxic waters and sediments, and it may contribute significantly to oceanic nitrogen budgets.
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            The anaerobic degradation of organic matter in Danish coastal sediments: iron reduction, manganese reduction, and sulfate reduction.

            We used a combination of porewater and solid phase analysis, as well as a series of sediment incubations, to quantify organic carbon oxidation by dissimilatory Fe reduction, Mn reduction, and sulfate reduction, in sediments from the Skagerrak (located off the northeast coast of Jutland, Denmark). In the deep portion of the basin, surface Mn enrichments reached 3.5 wt%, and Mn reduction was the only important anaerobic carbon oxidation process in the upper 10 cm of the sediment. In the less Mn-rich sediments from intermediate depths in the basin, Fe reduction ranged from somewhat less, to far more important than sulfate reduction. Most of the Mn reduction in these sediments may have been coupled to the oxidation of acid volatile sulfides (AVS), rather than to dissimilatory reduction. High rates of metal oxide reduction at all sites were driven by active recycling of both Fe and Mn, encouraged by bioturbation. Recycling was so rapid that the residence time of Fe and Mn oxides, with respect to reduction, ranged from 70-250 days. These results require that, on average, an atom of Fe or Mn is oxidized and reduced between 100-300 times before ultimate burial into the sediment. We observed that dissolved Mn2+ was completely removed onto fully oxidized Mn oxides until the oxidation level of the oxides was reduced to about 3.8, presumably reflecting the saturation by Mn2+ of highly reactive surface adsorption sites. Fully oxidized Mn oxides in sediments, then, may act as a cap preventing Mn2+ escape. We speculate that in shallow sediments of the Skagerrak, surface Mn oxides are present in a somewhat reduced oxidation level (< 3.8) allowing Mn2+ to escape, and perhaps providing the Mn2+ which enriches sediments of the deep basin.
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              N2 production by the anammox reaction in the anoxic water column of Golfo Dulce, Costa Rica.

              In oxygen-depleted zones of the open ocean, and in anoxic basins and fjords, denitrification (the bacterial reduction of nitrate to give N2) is recognized as the only significant process converting fixed nitrogen to gaseous N2. Primary production in the oceans is often limited by the availability of fixed nitrogen such as ammonium or nitrate, and nitrogen-removal processes consequently affect both ecosystem function and global biogeochemical cycles. It was recently discovered that the anaerobic oxidation of ammonium with nitrite--the 'anammox' reaction, performed by bacteria--was responsible for a significant fraction of N2 production in some marine sediments. Here we show that this reaction is also important in the anoxic waters of Golfo Dulce, a 200-m-deep coastal bay in Costa Rica, where it accounts for 19-35% of the total N2 formation in the water column. The water-column chemistry in Golfo Dulce is very similar to that in oxygen-depleted zones of the oceans--in which one-half to one-third of the global nitrogen removal is believed to occur. We therefore expect the anammox reaction to be a globally significant sink for oceanic nitrogen.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                4 September 2012
                : 7
                : 9
                : e42810
                Affiliations
                [1 ]AIMS@JCU, School of Marine and Tropical Biology, Australian Institute of Marine Science, Centre for Sustainable Tropical Fisheries and Aquaculture, James Cook University, Townsville, Queensland, Australia
                [2 ]School of Environmental Science and Management, Centre for Coastal Biogeochemistry, Southern Cross University, Lismore, New South Wales, Australia
                [3 ]Australian Institute of Marine Science, Townsville, Queensland, Australia
                [4 ]School of Marine, Tropical Biology and Centre for Sustainable Tropical Fisheries and Aquaculture, James Cook University, Townsville, Queensland, Australia
                Odum School of Ecology, University of Georgia, United States of America
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: SC DE LT NP. Performed the experiments: SC DE. Analyzed the data: SC DE NP. Contributed reagents/materials/analysis tools: SC DE BE. Wrote the paper: SC DE LT RdN.

                Article
                PONE-D-12-10160
                10.1371/journal.pone.0042810
                3433476
                22962581
                b190eda6-a509-4d5b-a315-0d8a52bf7e3d
                Copyright @ 2012

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 9 April 2012
                : 12 July 2012
                Page count
                Pages: 9
                Funding
                This research was funded by an AIMS@ 123456JCU research scholarship awarded to Sarah Castine at the beginning of her PhD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Agriculture
                Aquaculture
                Biology
                Biochemistry
                Metabolism
                Nitrogen Metabolism
                Chemistry
                Environmental Chemistry
                Marine Chemistry
                Pollutants
                Water Chemistry
                Geochemistry
                Biogeochemistry
                Engineering
                Environmental Engineering
                Pollution
                Water Management

                Uncategorized
                Uncategorized

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