Enhanced biohydrogen generation from organic wastewater containing NH 4 + by phototrophic bacteria Rhodobacter sphaeroides AR-3

NH ₄ ⁺ is typically an inhibitor to hydrogen production from organic wastewater by photo-bacteria. In this experiment, biohydrogen generation with wild-type anoxygenic phototrophic bacterium Rhodobacter sphaeroideswas found to be sensitive to NH ₄ ⁺ due to the significant inhibition of NH ₄ ⁺ to its nitrogenase. In order to avoid the inhibition of NH ₄ ⁺ to biohydrogen generation by R. sphaeroides, a glutamine auxotrophic mutant R. sphaeroides AR-3 was obtained by mutagenizing with ethyl methane sulfonate. The AR-3 mutant could generate biohydrogen efficiently in the hydrogen production medium with a higher NH ₄ ⁺ concentration, because the inhibition of NH ₄ ⁺ to nitrogenase of AR-3 was released. Under suitable conditions, AR-3 effectively produced biohydrogen from tofu wastewater, which normally contains 50–60 mg/L NH ₄ ⁺ , with an average generation rate of 14.2 mL/L·h. This generation rate was increased by more than 100% compared with that from wild-type R. sphaeroides.     

The role of metabolic nitrogen in coral calcification

When pieces of the staghorn coral Acropora acuminata are incubated with ¹⁴C-urea, the label is incorporated into skeletal carbonate. Incorporation of this label differs from that of H¹⁴CO ₃ ^- , suggesting urea is not immediately hydrolysed to provide a further source of HCO ₃ ^- . The effects of certain organic substrates upon calcification suggest the ornithine cycle is involved. Citrulline, an ornithine cycle intermediate, is found in high concentrations in the tissues of hermatypic corals. Urea, allantoins, NH₃ and arginine are also present. These compounds are barely detectable in zooxanthellae or an ahermatypic coral. The allantoins may be present as calcium salts. It is suggested that allantoins are the medium by which Ca²⁺ and CO₂ are transported to sites of calcification. Hydrolysis of urea, formed by breakdown of allantoins, yields CO₂ and NH₃. The NH₃ may neutralise protons formed during precipitation of CaCO₃ and bring about their removal from sites of calcification. As well as providing urea, the ornithine cycle may also be involved in the removal of NH₃ from sites of calcification.     

Experimental determination of Cd2+ adsorption mechanism on low-cost biological waste

Carbonate shells have an astonishing ability in the removal of Cd²⁺ in a short time period with emphasis on being a low cost adsorbent. In the present study, the sorption capacity of carbonate shells was studied for Cd²⁺ in batch experiments. The influence of different carbonate shell sizes and physico-chemical factors were evaluated and the results were analyzed for its correlation matrices by using Predictive Analytics Software (PASW). The mineralogy state of aqueous solution regarding the saturation index was simulated using PHREEQC to identify the Cd²⁺ uptake mechanism. The Cd uptake rates were calculated as well as Ca²⁺, HCO ₃ ^? concentration, pH, ambient humidity and temperature were measured. Cd²⁺ removal of 91.52% was achieved after 5 h adsorption. The adsorption efficiencies were significantly influenced by pH as they increased with the increase of pH from acidic solution (5.50±0.02) to slightly alkaline (7.60±0.05). In addition, the mineralogy state of aqueous solution calculated from PHREEQC confirmed that the increment of Ca²⁺ and HCO ₃ ^? concentrations in solution was attributed to the dissolution of carbonate shells. Moreover, the ion exchange adsorption mechanism of Cd²⁺ toward Ca²⁺ was identified as the process involved in Cd²⁺ uptake.     

Ammonium regeneration and carbon utilization by marine bacteria grown on mixed substrates

We examined the impact of exposing natural populations of marine bacteria (from seawater collected near Woods Hole, Massachusetts, USA) to multiple nitrogen and carbon sources in a series of batch growth experiments conducted from 1989 through 1990. The substrate C:N ratio (C:N_s) was varied from 1.5:1 to 10:1 either with equal amounts of NH ₄ ⁺ and different amino acids or an amino acid mixture, all supplemented with glucose to maintain the C:N_s ratio equal to that of the respective amino acid, or with combinations of glucose and NH ₄ ⁺ alone. A common feature of the experiments involving amino acids was the concurrent uptake of NH ₄ ⁺ and amino acids that persisted as long as a readily assimilable carbon source (glucose in our case) was taken up. There was no net regeneration of NH ₄ ⁺ , even though catabolism of amino acids occurred. Regeneration of NH ₄ ⁺ was evident only after glucose was completely utilized, which usually occurred at the end of exponential growth. The contribution of¹⁵NH ₄ ⁺ to total nitrogen uptake by the end of exponential growth varied from ~60 to 80% when individual amino acids were present and down to ~24% when the amino acid mixture was added. These estimates are conservative because we did not account for possible isotope dilution effects resulting from amino acid catabolism. When NH ₄ ⁺ and glucose were the sole nitrogen and carbon sources, there was a stoichiometric balance between glucose and NH ₄ ⁺ uptake over a wide range of C:N_s ratios, leading to a constant bacterial biomass C:N ratio (C:N_B) of ~4.5:1. As a result NH ₄ ⁺ usage varied from 50% when the C:N_s ratio was 3.6:1, to 100% when the C:N_s ratio was 10:1. Gross growth efficiency varied from ~60% when NH ₄ ⁺ plus glucose were added alone or with the amino acid mixture, to 47% when the individual amino acids were used in place of the mixture. It is thus evident that actively growing bacteria will act as sinks for nitrogen when a carbon source that can be assimilated easily is available to balance NH ₄ ⁺ uptake, even when amino acids are available and are being co-metabolized.     

Nitrate uptake in marine phytoplankton: Energy sources and the interaction with carbon fixation

Field studies of whole natural phytoplankton communities from Knight Inlet, B. C., Canada and laboratory cultures of the diatom Skeletonema costatum indicate inorganic carbon fixation may be temporarily suppressed following 10 to 15% enrichment with NO ₃ ^- or NH ₄ ⁺ . (This effect is suggested to be due to competition between inorganic carbon and nitrogen for adenosine triphosphate (ATP), and is reduced when chlorophyll a is increased intracellularly after 6 to 8 h.) Results imply that the source of ATP for nitrate uptake is primarily from Photosystem I (cyclic photophosphorylation) in the presence of light. It would appear that a transient nutrient-adaptive response occurs upon addition of extracellular nitrogen.     

Short-term effects of excessive anaerobic reaction time on anaerobic metabolism of denitrifying polyphosphate-accumulating organisms linked to phosphorus removal and N2O production

The short-term effect of anaerobic reaction time (AnRT) (i.e., 90, 120 and 150 min) on the denitrifying phosphorus (P) removal performance and N₂O production was examined using a denitrifying enhanced biologic phosphorus removal (EBPR) sludge acclimatized with mixed acetate (HAc) and propionate (Pro) (in the molar ratio 3:1) as carbon sources. The results showed that when the AnRT was prolonged from 90 to 150 min, the anaerobic polyhydroxyalkanoate (PHA) synthesis was decreased by 15.3%. Moreover, the ineffective PHA consumption occurred in anaerobic phases and contributed to an increased NO ₂ ^? -N accumulation and higher free nitrous acid (FNA) concentrations (?0.001–0.0011 mg HNO₂-N/L) in the subsequent anoxic phases, causing a severe inhibition on anoxic P-uptake and denitrification. Accordingly, the total nitrogen (TN) and total phosphorus (TP) removal efficiencies dropped by approximately 6.3% and 85.5%, respectively; and the ratio of anoxic N₂O-N production to TN removal increased by approximately 3.8%. The fluorescence in situ hybridization (FISH) analysis revealed that the sludge was mainly dominated by Accumulibacter (62.0% (SE_mean = 1.5%)). In conclusion, the short-term excessive anaerobic reaction time negatively impacted denitrifying P removal performance and stimulated more N₂O production, and its effect on P removal was more obvious than that on nitrogen removal.     

Kinetics of the oxidation of endocrine disruptor nonylphenol by ferrate(VI)

The kinetics of the oxidation of endocrine disruptor nonylphenol (NP) by potassium ferrate(VI) (K₂FeO₄) in water as a function of pH 8.0–10.9 at 25°C is presented. The observed second-order rate constants, k _obs, decrease with an increase in pH 269–32 M^?1 s^?1. The speciation of Fe(VI) (HFeO ₄ ^? and FeO ₄ ^2? ) and NP (NP–OH and NP–O^?) species was used to explain the pH dependence of the k _obs values. At a dose of 10 mg L^?1 (50 μM) K₂FeO₄, the half-life for the removal of NP by Fe(VI), under water treatment conditions, is less than 1 min.     

Effect of nitrogen source on short-term light and dark CO2 uptake by a marine diatom

Based on a series of short-term incubations involving the marine diatom Chaetoceros simplex (Bbsm), precultured in NH ₄ ⁺ -, NO ₃ ^- -and urea-limited continuous cultures at several dilution rates, we found that both the short-term specific rate of ¹⁴CO₂ uptake and the amount of CO₂ fixed after 8- and 16-min incubations were unaffected by enrichment with NH ₄ ⁺ , urea, or NO ₃ ^- when NH ₄ ⁺ or urea were the preconditioning forms of N, but were slightly suppressed when the cells were first grown on NO ₃ ^- . Similar enrichments in the dark, however, led to significant CO₂ uptake under all conditions of NH ₄ ⁺ enrichment and to similarly enhanced CO₂ uptake, but only at high growth rates, when urea was the source of enrichment nitrogen. Our light results are contrary to some contemporary findings, but there does seem to be agreement that photosynthetic rates of rapidly growing phytoplankton will not be affected by exposure to pulses of nitrogen. Enhanced dark uptake, in contrast, appears to be characteristic of phytoplankton under all degrees of N limitation, and, as such, may be useful as an “all or nothing” index of the nitrogen status of natural waters. There is some indication that the index may be useful in determining both the form of and the degree of N limitation as well.     

Urea as a nitrogen source for the phytoplankton in the Oslofjord

Ambient concentrations of urea in the inner Oslofjord, Norway, showed a pronounced yearly cycle in 1980, with values in the range 0.1 to 10.0 μg-at N l^-1; this cycle resemble that of ammonia although urea concentrations were usually lower. The uptake of urea by phytoplankton was investigated using ¹⁵N. Urea was usually a less important N source than NH ₄ ⁺ , and accounted for 0 to 53% (mean 19%) of summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rates from April to October. Absolute as well as relative (specific) uptake rates of urea were higher in the summer (June–August) than at other times. Uptake of urea was inhibited by NH ₄ ⁺ concentrations higher than 1 to 2 μg-at N l^-1. The summed NH ₄ ⁺ +NO ₃ ^- + urea uptake rate was exponentially related to temperature.     

Use of zeolitized coal fly ash in the simultaneous removal of ammonium and phosphate from aqueous solution

Discharge of wastewater containing nitrogen and phosphate can cause eutrophication. Therefore, the development of an efficient material for the immobilization of the nutrients is important. In this study, a low calcium fly ash and high calcium fly ash were converted into zeolite using the hydrothermal method. The removal of ammonium and phosphate that coexist in aqueous solution by the synthesized zeolites were studied. The results showed that zeolitized fly ash could efficiently eliminate ammonium and phosphate at the same time. Saturation of zeolite with Ca²⁺ rather than Na⁺ favored the removal of both ammonium and phosphate because the cation exchange reaction by the NH₄ ⁺ resulted in the release of Ca²⁺ into the solution and precipitation of Ca²⁺ with PO₄ ^3? followed. An increase in the temperature elevated the immobilization of phosphate whereas it abated the removal of ammonium. Nearly 60% removal efficiency for ammonium was achieved in the neutral pH range from 5.5 to 10.5, while the increase or decrease in pH out of the neutral range lowered the adsorption. In contrast, the removal of phosphate approached 100% at a pH lower than 5.0 or higher than 9.0, and less phosphate was immobilized at neutral pH. However, there was still a narrow pH range from 9.0 to 10.5 favoring the removal of both ammonium and phosphate. It was concluded that the removal of ammonium was caused by cation exchange; the contribution of NH₃ volatilization to immobilization at alkaline conditions (up to pH level of 11.4) was limited. With respect to phosphate immobilization, the mechanism was mainly the formation of precipitate as Ca₃(PO₄)₂ within the basic pH range or as FePO₄ and AlPO₄ within acidic pH range.     

Performance of completely autotrophic nitrogen removal over nitrite process under different aeration modes and dissolved oxygen

In this study, three sequential batch biofilm reactors (SBBRs) were operated for 155 days to evaluate the performance of completely autotrophic nitrogen removal over nitrite (CANON) process under different aeration modes and dissolved oxygen (DO). Synthetic wastewater with 160-mg NH4 ⁺-N/L was fed into the reactors. In the continuously-aerated reactor, the efficiency of the ammonium nitrogen conversion and total nitrogen (TN) removal reached 80% and 70%, respectively, with DO between 0.8–1.0 mg/L. Whereas in the intermittently-aerated reactor, at the aeration/non-aeration ratio of 1.0, ammonium was always under the detection limit and 86% of TN was removed with DO between 2.0–2.5 mg/L during the aeration time. Results show that CANON could be achieved in both continuous and intermittent aeration pattern. However, to achieve the same nitrogen removal efficiency, the DO needed in the intermittently-aerated sequential batch biofilm reactor (SBBR) during the aeration period was higher than that in the continuously-aerated SBBR. In addition, the DO in the CANON system should be adjusted to the aeration mode, and low DO was not a prerequisite to CANON process.     

Comparison of different algicides on growth of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> and microcystin release,as well as its removal pathway in riverways

Eutrophication with a large number of Microcystis aeruginosa commonly occurs worldwide, thereby threatening the aquatic ecosystem and human health. In this study, four kinds of algicides were tested to explore their influence on cell density and chlorophyll-a of M. aeruginosa. Results showed that aluminum silicate agent, which inhibited more than 90% cell growth compared with the control group, demonstrated the strongest inhibition effect immediately on M. aeruginosa growth. Furthermore, the production and release of microcystin (MC)-LR were investigated. Aluminum silicate, CuSO₄, and Emma-11 were more effective than pyrogallic acid in disrupting the cells of M. aeruginosa, thereby increasing the extracellular MC-LR concentration. Aluminum silicate caused the highest extracellular MC-LR concentration of more than 45 mg·L^–1. Biotoxicity was also detected to evaluate the environmental risks of MC-LR release, which were related to the usages of different algicides. Extracellular MC-LR concentration mostly increased when the biotoxicity of algae solution increased. The experiments were also designed to reveal the effects of physical conditions in riverways, such as natural sunlight, aeration and benthal sludge, on MC-LR degradation. These findings indicated that UV rays in sunlight, which can achieve a MC-LR removal efficiency of more than 15%, played an important role in MC-LR degradation. Among all the physical pathways of MC-LR removal, benthal sludge adsorption presented the optimal efficiency at 20%.

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Arsenic (V) removal from groundwater by GE-HL nanofiltration membrane: effects of arsenic concentration, pH, and co-existing ions

A laboratory-scale investigation was performed to study arsenic (As (V)) removal by negatively charged GE-HL nanofiltration (NF) membrane in simulated drinking water. Effects of As (V) concentration (0–200 μg·L^?1), pH, and co-ions and counter-ions were investigated. The NF membrane presented good stability, and the rejection rates exceeded 90%. The rejection rates of As (V) decreased with the increase of As (V) concentration, while it increased with the increase of pH (reached 96% at pH 6.75). Moreover, a negative relationship was observed between the co-existing ions of Cl^?, Na⁺, SO ₄ ^2? , and Ca²⁺ and the removal of As (V), in which bivalent ions presented more significant effects than monovalent ions.     

Metabolism of two dominant epibenthic echinoderms measured at bathyal depths in the Santa Catalina Basin

Metabolism of two abundant echinoderm species constituting 99.6% of the epibenthic megafauna in the Santa Catalina Basin, off southern California, USA was measured at 1 300 m during the 1979 “Bathyal Expedition”. Specimens of the ophiuroid Ophiophthalmus normani and the holothurian Scotoplanes globosa, collected by the submersible “Alvin”, were individually placed in respirometers, and measured in situ for O₂ consumption and ammonium excretion rates. For O. normani, weight-specific O₂ consumption rates decreased with increasing weight and were of comparable magnitude to rates of deep-sea and shallow-water ophiuroids; excretion rates were highly variable. Population O₂ consumption and excretion rates for O. normani (estimated from size-frequency distribution, abundance, and rate regression equations) were 1 129.28 μl O₂ m^-2 h^-1 and 27.30 nmol NH ₄ ⁺ m^-2 h^-1. Weight-specific O₂ consumption and ammonium excretion rates of S. globosa decreased with increasing weight and were of comparable magnitude to rases of shallow-water holothurians. Population O₂ constimption and excretion rates of S. globosa were 1.38 μl O₂ m^-2 h^-1 and 4.86 nmol NH ₄ ⁺ m^-2 h^-1. Combined population O₂ consumption rates for O. normani and S. globosa are of comparable magnitude to that of the sediment community and plankton in the benthic boundary layer (sediment and overlying 50 m water column) of the Santa Catalina Basin.     

Klebsiella pneumoniae metabolites enhance Microcystis aeruginosa biomass flocculation

Microcystis aeruginosa is a common aquatic toxin-producing cyanobacterium. This study explores M. aeruginosa biomass recovery using bioflocculation. Bioflocculants produced by Klebsiella pneumoniae significantly enhanced aquatic M. aeruginosa flocculation efficiency. Under neutral and acidic conditions, flocculation efficiency reached 90%, whereas flocculation efficiency was 75% under alkaline conditions. M. aeruginosa flocculation was ion dependent, and Ca²⁺ was determined to be the most effective cationic inducer. Negatively charged proteoglycans were identified as the being the functional components in the flocculants. Optimal conditions for M. aeruginosa flocculation were determined using response surface methodology and are as follows: (1) K. pneumoniae bioflocculant dose, 13.3?mg/L; (2) flocculation time, 1.6 hours; and (3) pH?=?5 for flocculation of 10⁹ cells per litre. Bioflocculation using K. pneumoniae represents a promising method to control development of M. aeruginosa blooms. Further in-field investigations are necessary to fully assess its potential.     

Removing nitrogen and phosphorus from simulated wastewater using algal biofilm technique

Algal biofilmtechnology is a new and advanced wastewater treatment method. Experimental study on removing nitrogen and phosphorus from simulated wastewater using algal biofilm under the continuous light of 3500 Lux in the batch and continuous systems was carried out in this paper to assess the performance of algal biofilm in removing nutrients. The results showed that the effect of removing nitrogen and phosphorus by algal biofilm was remarkable in the batch system. The removal efficiencies of total phosphorus (TP), total nitrogen (TN), ammonia-nitrogen (NH₃-N), and chemical oxygen demand (COD) reached 98.17%, 86.58%, 91.88%, and 97.11%, respectively. In the continuous system, hydraulic retention time (HRT) of 4 days was adopted; the effects of removing TP, TN, NH₃-N, and COD by algal biofilm were very stable. During a run of 24 days, the removal efficiencies of TP, TN, NH₃-N, and COD reached 95.38%, 83.93%, 82.38%, and 92.31%, respectively. This study demonstrates the feasibility of removing nitrogen and phosphorus from simulated wastewater using algal biofilm.     

Fate and effects of nonylphenol in the presence of the cyanobacterium Microcystis aeruginosa

Interactions of nonylphenol (NP), a toxic and oestrogenic degradation product of widely used non-ionic surfactants, with the cyanobacterium Microcystis aeruginosa were studied. Batch cultures were incubated for 10 days with NP concentrations between 10 and 570 nM. NP was removed more quickly in the presence of M. aeruginosa (half-life 2.7–5.2 days) than in its absence (half-life 6.7–10.2 days) at all concentrations tested. At the end of the experiment, NP could not be found in the biomass, so the biotic removal is due to uptake and chemical transformation, and not to physical binding on the cells. The observed effective concentrations, EC₅₀ and EC₂₀, were 0.45 and 0.25 μ M, respectively. Therefore, NP is expected to have toxic effects on M. aeruginosa only in very contaminated surface waters. However, for concentrations that go far beyond environmental levels, cyanobacteria are able to cope with NP toxicity by internalising the compound in a less toxic form. Therefore, the presence of cyanobacteria may increase the rate of NP removal from the aquatic environment.     

Microbial interactions with the cyanobacterium Microcystis aeruginosa and their dependence on temperature

Associated heterotrophic bacteria alter the microenvironment of cyanobacteria and potentially influence cyanobacterial development. Therefore, we studied interactions of the unicellular freshwater cyanobacterium Microcystis aeruginosa with heterotrophic bacteria. The associated bacterial community was greatly driven by temperature as seen by DNA fingerprinting. However, the associated microbes also closely interacted with the cyanobacteria indicating changing ecological consequence of the associated bacterial community with temperature. Whereas concentration of dissolved organic carbon in cyanobacterial cultures changed in a temperature-dependent manner, its quality greatly varied under the same environmental conditions, but with different associated bacterial communities. Furthermore, temperature affected quantity and quality of cell-bound microcystins, whereby interactions between M. aeruginosa and their associated community often masked this temperature effect. Both macro- and microenvironment of active cyanobacterial strains were characterized by high pH and oxygen values creating a unique habitat that potentially affects microbial diversity and function. For example, archaea including ‘anaerobic’ methanogens contributed to the associated microbial community. This implies so far uncharacterized interactions between Microcystis aeruginosa and its associated prokaryotic community, which has unknown ecological consequences in a climatically changing world.     

Phytoplankton dynamics and species diversity in a shallow eutrophic,natural mid-altitude lake in Himachal Pradesh (India): role of physicochemical factors

Rewalsar Lake, a mid-altitude, shallow and recreational water body located in the north-western Himalayas, Himachal Pradesh (India) was studied through monthly surveys in two consecutive years (March 2008 to February 2010). Forty-seven species belonging to seven groups of phytoplankton were identified from the lake. Microcystis aeruginosa and Synedra ulna exhibited a perennial habit. Ankistrodesmus falcatus, Chlorella vulgaris, Scenedesmus bijugatus, Chlamydomonas reinhardi, Eudorina elegans, Navicula cuspidate, Synedra ulna, Euglena acus, Euglena oxyuris, Spirulina gomontii, Oscillatoria princeps and Arthrospira khannae were abundant, and Oscillatoria limosa and Microcystis aeruginosa were highly abundant. Twenty-one important criteria were studied, for example, temperature, free carbon dioxide, biochemical oxygen demand, total alkalinity, nitrate, silicate and phosphate, which provide an idea of the portability of water for irrigation and drinking purposes as per the permissible limits given in World Health Organization, Indian Council of Medical Research and Indian Standards Institute standards. Pearson's correlation revealed a significant relationship between physicochemical parameters and different algal groups. Both plankton and chlorophyll a showed a bimodal pattern of fluctuation. High annual mean concentrations of chlorophyll a (mg L^?1) were recorded as 11.44 in 2008–09, and 11.04 in 2009–10. As per the Palmer pollution index, 13 pollution-tolerant algal species with a pollution score of 37 were observed. The Central Pollution Control Board categorised the water at Rewalsar Lake as ‘D–E’.