Phosphate Uptake Rates Research Articles

Identification of 2, 4-di-tert-butylphenol from Microcystis lysate after bloom control and its potential risks to aquatic ecosystems

With the increasing concern of cyanobacterial blooms, numerous techniques have been developed to mitigate these environmental nuisances. During bloom control, the allelopathic effects of compounds released from cyanobacterial cells are considered as secondary hazards. In this study, the findings indicated that Microcystis lysate inhibited algal proliferation and disrupted the development of zebrafish embryos. Then, allelochemicals in Microcystis lysate were identified using gas chromatography-mass spectrometry, with 2, 4-di-tert-butylphenol (2, 4-DTBP) being the only identified phenol, which was selected for further study. The results showed that 2,4-DTBP caused oxidative damages, disrupted metabolic activity, and suppressed photosynthetic activity, consequently impeding the growth of Microcystis aeruginosa (M. aeruginosa) and Chlorella pyrenoidosa (C. pyrenoidosa). Moreover, it enhanced the interspecies competitive advantages of M. aeruginosa by increasing phosphate uptake rate. Furthermore, at a concentration of 2 mg L−1, 2, 4-DTBP negatively affected the development of zebrafish embryos, manifesting in mortality, malformation, and hatching delay. Therefore, the investigation identified 2, 4-DTBP as a potential allelochemical within Microcystis lysate. Although the effective concentration for freshwater algae and zebrafish embryos was higher than that found in Microcystis lysate, it highlighted the need for careful monitoring of aquatic ecosystem health during cyanobacterial bloom mitigation.

Influence of operation sequences on phosphorus recovery by polyphosphate-accumulating organisms biofilm: Performance, kinetics and metabolic response

Two biofilm sequencing batch reactors (BSBRs) were conducted under alternating aerobic/anaerobic (Ae/An) sequence and alternating anaerobic/aerobic (An/Ae) sequence to investigate the effects of operation sequences on phosphorus removal and recovery performance. The Ae/An BSBR had advantages in phosphate enrichment, phosphorus concentration of enrichment solution can reach 99.08 mg/L. While An/Ae BSBR exhibited a higher kinetic rate for phosphate uptake and release up to 0.48 mmol/gVSS. In An/Ae BSBR, the remarkable increase of phosphate concentration in aerobic stage put a great strain on the phosphate uptake of biofilms that led to an increase of phosphorus accumulation in the biofilm (Pbiofilm). However, the poly-P proportion in TPbiofilm decreased which triggered the metabolisim of polyphosphate accumulating organisms (PAOs) shifting from polyphosphate accumulating metabolism (PAM) to glycogen accumulating metabolism (GAM) and suppressed the phosphorus enrichment performance of the An/Ae BSBR. The phosphate uptake rate (PUR) of biofilms was in alignment with the bulk phosphate concentration. PAOs were dominated by Rhodoplanes and Saprospiraceae in the Ae/An BSBR, whereas Thiothrix was predominant in the An/Ae BSBR. Notably, the Candidatus Competibacter, considered as a denitrifying glycogen accumulating organisms (DGAOs), showed a dominant proportion in the microbial community of both BSBRs. The higher expression of actP in the An/Ae system indicated a greater capacity for the uptake of acetate, while the Ae/An system was is more adept for degrading acetate represented by the higher relative abundance of ackA, pta and acsA genes The high phosphorus loading in the An/Ae system suppressed the ppk expression but enhanced the ppx expression.

Effect of an NHE3 inhibitor in combination with an NPT2b inhibitor on gastrointestinal phosphate absorption in Rodent models.

Many of the pathological consequences of chronic kidney disease can be attributed to an elevation in serum phosphate levels. Current therapies focused on decreasing intestinal phosphate absorption to treat hyperphosphatemia are inadequate. The most effective therapeutic strategy may be to target multiple absorptive pathways. In this study, the ability of a novel inhibitor of the intestinal sodium hydrogen exchanger 3 (NHE3), LY3304000, which inhibits paracellular, diffusional uptake of phosphate, to work in combination with an inhibitor of the active transporter, sodium dependent phosphate cotransporter 2b (NPT2b), LY3358966, was explored. LY3304000 modestly inhibited the acute uptake of phosphate into plasma of rats, while surprisingly, it doubled the rate of phosphate uptake in mice, an animal model dominated by NPT2b mediated acute phosphate uptake. In rats, LY3004000 and LY3358966 work in concert to inhibit acute phosphate uptake. On top of LY3358966, LY3304000 further decreased the acute uptake of phosphate into plasma. Studies measuring the recovery of radiolabeled phosphate in the intestine demonstrated LY3304000 and LY3358966 synergistically inhibited the absorption of phosphate in rats. We hypothesize the synergism is because the NHE3 inhibitor, LY3304000, has two opposing effects on intestinal phosphate absorption in rats, first it decreases diffusion mediated paracellular phosphate absorption, while second, it simultaneously increases phosphate absorption through the NPT2b pathway. NHE3 inhibition decreases proton export from enterocytes and raises the cell surface pH. In vitro, NPT2b mediated phosphate transport is increased at higher pHs. The increased NPT2b mediated transport induced by NHE3 inhibition is masked in rats which have relatively low levels of NPT2b mediated phosphate transport, by the more robust inhibition of diffusion mediated phosphate absorption. Thus, the inhibition of NPT2b mediated phosphate transport in rats in the presence of NHE3 inhibition has an effect that exceeds its effect in the absence of NHE3 inhibition, leading to the observed synergism on phosphate absorption between NPT2b and NHE3 inhibition.

Godzilla mineral dust and La Soufrière volcanic ash fallout immediately stimulate marine microbial phosphate uptake

During the “Godzilla” dust storm of June 2020, unusually high fluxes of mineral dust traveled across the Atlantic from the Sahara Desert, reaching the Caribbean Basin, Gulf Coast, and southeastern United States. Additionally, an eruption of the La Soufrière volcano on St. Vincent in April 2021 generated substantial ashfall in the southeastern Caribbean. While many studies have analyzed mineral dust’s ability to relieve nutrient limitation of phosphorus (P) in the P-stressed North Atlantic, less is known about the impact of extreme events and other natural aerosols on fluxes of P into seawater and from seawater into marine microbial cells. We quantified P and iron (Fe) content in mineral dust from the Godzilla dust storm and volcanic ash from the La Soufrière eruption collected at Ragged Point, Barbados. We also performed seawater incubations to assess the marine microbial response to aerosol deposition. Using environmentally-relevant concentrations of atmospheric particles for within the ocean’s mixed layer allowed us to draw realistic conclusions about how these deposition events impacted P cycling in situ. Volcanic ash has lower P content than mineral dust, and P in volcanic ash is far less soluble (~1%) than assumed in current atmospheric deposition models. Adding mineral dust and the volcanic ash leachate in concentrations representing different deposition scenarios increased soluble reactive phosphorus (SRP) concentrations in coastal seawater by ~7-32 nM. Phosphate uptake rate was stimulated in coastal seawater after either mineral dust or volcanic ash deposition at aerosol concentrations relevant to the Godzilla dust event, with ash eliciting the fastest uptake rate. Furthermore, high concentrations of both the mineral dust and volcanic ash led to slightly elevated alkaline phosphatase activity (APA) compared to the relevant controls, indicating higher potential for use of dissolved organic phosphorus (DOP) as a P source. Quantifying these aerosols’ impacts on P cycling is a significant step towards achieving a better understanding of their potential roles in relieving nutrient limitation and fueling the biological carbon pump.

Low cobalt inventories in the Amundsen and Ross seas driven by high demand for labile cobalt uptake among native phytoplankton communities

Abstract. Cobalt (Co) is a scarce but essential micronutrient for marine plankton in the Southern Ocean and coastal Antarctic seas, where dissolved cobalt (dCo) concentrations can be extremely low. This study presents total dCo and labile dCo distributions measured via shipboard voltammetry in the Amundsen Sea, the Ross Sea and Terra Nova Bay during the CICLOPS (Cobalamin and Iron Co-Limitation of Phytoplankton Species) expedition. A significantly smaller dCo inventory was observed during the 2017/2018 CICLOPS expedition compared to two 2005/2006 expeditions to the Ross Sea conducted over a decade earlier. The dCo inventory loss (∼ 10–20 pM) was present in both the surface and deep ocean and was attributed to the loss of labile dCo, resulting in the near-complete complexation of dCo by strong ligands in the photic zone. A changing dCo inventory in Antarctic coastal seas could be driven by the alleviation of iron (Fe) limitation in coastal areas, where the flux of Fe-rich sediments from melting ice shelves and deep sediment resuspension may have shifted the region towards vitamin B12 and/or zinc (Zn) limitation, both of which are likely to increase the demand for Co among marine plankton. High demand for Zn by phytoplankton can result in increased Co and cadmium (Cd) uptake because these metals often share the same metal uptake transporters. This study compared the magnitudes and ratios of Zn, Cd and Co uptake (ρ) across upper-ocean profiles and the observed order-of-magnitude uptake trends (ρZn > ρCd > ρCo) that paralleled the trace metal concentrations in seawater. High rates of Co and Zn uptake were observed throughout the region, and the speciation of available Co and Zn appeared to influence trends in dissolved metal : phosphate stoichiometry and uptake rates over depth. Multi-year loss of the dCo inventory throughout the water column may be explained by an increase in Co uptake into particulate organic matter and subsequently an increased flux of Co into sediments via sinking and burial. This perturbation of the Southern Ocean Co biogeochemical cycle could signal changes in the nutrient limitation regimes, phytoplankton bloom composition and carbon sequestration sink of the Southern Ocean.

Chemical mutagenesis and thermal selection of coral photosymbionts induce adaptation to heat stress with trait trade-offs.

Despite the relevance of heat-evolved microalgal endosymbionts to coral reef restoration, to date, few Symbiodiniaceae strains have been thermally enhanced via experimental evolution. Here, we investigated whether the thermal tolerance of Symbiodiniaceae can be increased through chemical mutagenesis followed by thermal selection. Strains of Durusdinium trenchii, Fugacium kawagutii and Symbiodinium pilosum were exposed to ethyl methanesulfonate to induce random mutagenesis, and then underwent thermal selection at high temperature (31/33°C). After 4.6-5 years of experimental evolution, the in vitro thermal tolerance of these strains was assessed via reciprocal transplant experiments to ambient (27°C) and elevated (31/35°C) temperatures. Growth, photosynthetic efficiency, oxidative stress and nutrient use were measured to compare thermal tolerance between strains. Heat-evolved D. trenchii, F. kawagutii and S. pilosum strains all exhibited increased photosynthetic efficiency under thermal stress. However, trade-offs in growth rates were observed for the heat-evolved D. trenchii lineage at both ambient and elevated temperatures. Reduced phosphate and nitrate uptake rates in F. kawagutii and S. pilosum heat-evolved lineages, respectively, suggest alterations in nutrition resource usage and allocation processes may have occurred. Increased phosphate uptake rates of the heat-evolved D.trenchii strain indicate that experimental evolution resulted in further trade-offs in this species. These findings deepen our understanding of the physiological responses of Symbiodiniaceae cultures to thermal selection and their capacity to adapt to elevated temperatures. The new heat-evolved Symbiodiniaceae developed here may be beneficial for coral reef restoration efforts if their enhanced thermal tolerance can be conferred in hospite.

Impact of wastewater characteristics on the removal of organic micropollutants by Chlorella sorokiniana

Microalgae-based technologies can be used for the removal of organic micropollutants (OMPs) from different types of wastewater. However, the effect of wastewater characteristics on the removal is still poorly understood. In this study, the removal of sixteen OMPs by Chlorella sorokiniana, cultivated in three types of wastewater (anaerobically digested black water (AnBW), municipal wastewater (MW), and secondary clarified effluent (SCE)), were assessed. During batch operational mode, eleven OMPs were removed from AnBW and MW. When switching from batch to continuous mode (0.8 d HRT), the removal of most OMPs from AnBW and MW decreased, suggesting that a longer retention time enhances the removal of some OMPs. Most OMPs were not removed from SCE since poor nutrient availability limited C. sorokiniana growth. Further correlation analyses between wastewater characteristics, biomass and OMPs removal indicated that the wastewater soluble COD and biomass concentration predominantly affected the removal of OMPs. Lastly, carbon uptake rate had a higher effect on the removal of OMPs than nitrogen and phosphate uptake rate. These data will give an insight on the implementation of microalgae-based technologies for the removal of OMPs in wastewater with varying strengths and nutrient availability.

Phosphorus uptake and accumulation in Chlamydomonas reinhardtii: Influence of biomass concentration, phosphate concentration, phosphorus depletion time, and light supply

The model microalga Chlamydomonas reinhardtii was cultivated under P depleted and P repleted conditions to test the influence of dissolved P concentration (0–20 mg-P·L−1), initial algal biomass concentration (0.17–0.57 g-DW·L−1), P-depletion time (1 h – 4 days) and light supply (continuous illumination versus darkness) on phosphate uptake rate (mg-P·h−1·L−1) and the cellular P content (as %P of dry cell mass). Dissolved P concentration positively influenced cellular P content, while biomass concentration and P-depletion time did not. Biomass concentration and P-depletion time instead positively influenced P uptake rates. Light supply neither influenced cellular P content nor P assimilation. These findings evidence that depleting the cells at high initial biomass concentration could enhance P removal during wastewater treatment in waste stabilization ponds. Practically, these findings imply that high P removal from wastewater could be achieved by exposing ‘concentrated’ P-depleted biomass to P-laden influent in a relatively short contact-time reactor and subsequently harvesting and removing the resultant P-rich biomass. This stage would be followed by normal biomass growth in larger ponds which depletes the remaining P, with this biomass being harvested and returned to the contact tank for P removal.

Uptake of excess phosphate at low inorganic N:P ratio in a coastal sea afflicted with eutrophication

Following the spring bloom in the northern Baltic Sea, nitrogen limits phytoplankton growth and there is typically a residual phosphate concentration (>0.2 µmol l-1) remaining that is often assumed to induce the recurring blooms of nitrogen-fixing cyanobacteria. However, these cyanobacterial blooms typically occur 2-3 mo later during the summer, when the phosphate concentration has been depleted, and it is unclear what organisms take up the excess phosphate. We studied the removal potential of excess phosphate (0.55 µmol l-1) at different temperatures (10, 13, 16°C) and with or without nitrogen addition in an indoor 20 l tank experiment. In addition, we followed the element pools and plankton community composition. As expected, the phosphate uptake rate was up to 3-fold faster in nitrogen-amended than non-amended tanks, but complete drawdown of phosphate also occurred under severe nitrogen limitation. The uptake ratio of dissolved inorganic nitrogen to phosphorus was 4.6, which is substantially lower than the Redfield ratio (16) and indicates excessive phosphate removal potential relative to nitrogen. A large part of the excess phosphate ended up in the particulate pool, which has a higher potential to sink out from the surface. The nitrogen-fixing cyanobacteria Nodularia spumigena grew close to summer bloom concentrations only in the highest experimental temperature. However, the combined biovolume of all 3 major bloom-forming cyanobacteria accounted for only 5.3% of the total autotropic biovolume, and their potential phosphate uptake was calculated to be <3% of the excess phosphate available at the beginning of the study. Therefore, our results demonstrate that the contribution of filamentous cyanobacteria to the removal of excess phosphate is small.

Unsuspected functions of alkaline phosphatase PhoD in the diatom Phaeodactylum tricornutum

Alkaline phosphatase (AP) in algae is known as the indicator of phosphorus nutrient (P) starvation and an enzyme to hydrolyze dissolved organic phosphorus (DOP) as an alternative source of P when the primary source of P, dissolved inorganic phosphate, DIP, is limited in the marine environment. Here, we document the roles of an intracellular PhoD type AP gene in the diatom Phaeodactylum tricornutum other than DOP scavenging when P supply is abundant. CRISPR-mediated mutation of PhoD_45757 resulted in increased lipid content, pigment content, photosynthetic efficiency, and oxygen evolution. Meanwhile, PhoD_45757 mutation also altered physiological and transcriptomic performance under different P conditions. Mutant strains grown in the P-replete medium exhibited significantly lower growth rates than the wild type, apparently by inhibiting G1 phase in the cell cycle, and higher phosphate uptake rate, accompanied by increased particulate P and RNA contents per cell. In addition, mPhoD cells increased in cellular carbon (including lipid) and nitrogen contents, and reprogrammed their metabolic response to changes in the nutrient condition. Our physiological and transcriptomic results reveal the roles of a PhoD in pigment synthesis and photosynthesis, lipid accumulation, cell cycle regulation, and nutrient homeostasis in a diatom when DOP scavenging is not required.

Biochemical and structural characterization of the cyanophage-encoded phosphate-binding protein: implications for enhanced phosphate uptake of infected cyanobacteria.

To acquire phosphorus, cyanobacteria use the typical bacterial ABC-type phosphate transporter, which is composed of a periplasmic high-affinity phosphate-binding protein PstS and a channel formed by two transmembrane proteins PstC and PstA. A putative pstS gene was identified in the genomes of cyanophages that infect the unicellular marine cyanobacteria Prochlorococcus and Synechococcus. However, it has not been determined whether the cyanophage PstS protein is functional during infection to enhance the phosphate uptake rate of host cells. Here we showed that the cyanophage P-SSM2 PstS protein was abundant in the infected Prochlorococcus NATL2A cells and the host phosphate uptake rate was enhanced after infection. This is consistent with our biochemical and structural analyses showing that the phage PstS protein is indeed a high-affinity phosphate-binding protein. We further modelled the complex structure of phage PstS with host PstCA and revealed three putative interfaces that may facilitate the formation of a chimeric ABC transporter. Our results provide insights into the molecular mechanism by which cyanophages enhance the phosphate uptake rate of cyanobacteria. Phosphate acquisition by infected bacteria can increase the phosphorus contents of released cellular debris and virus particles, which together constitute a significant proportion of the marine dissolved organic phosphorus pool.

Morpho-physiological adaptations of Leptocylindrus aporus and L. hargravesii to phosphate limitation in the northern Adriatic

The northern Adriatic is highly productive and shallow area characterized by numerous spatio-temporal gradients (e.g. nutrients, salinity, temperature). It is strongly influenced by numerous freshwater inputs, mainly from Po river. Its current systems as well as Po river, generates gradients of phosphate availability with an expressed N/P imbalance and phosphate limitation. A number of recent studies characterized these gradients as major factors affecting abundance and composition of microphytoplankton communities. Focus of this study is on two Leptocylindrus species, Leptocylindrus aporus (F.W. French & Hargraves) D. Nanjappa & A. Zingone 2013 and Leptocylindrus hargravesii D. Nanjappa & A. Zingone 2013. Species belonging to Leptocylindrus genus are frequently observed and have high abundances and also high contributions to the microphytoplankton community in this area. We focused on their morphological and physiological responses to phosphate limitation in situ and also performed in vitro experiments. In this study we report data on species specific growth rates under phosphorus (P) deplete and P rich conditions, localization and characteristics of alkaline phosphate activity, phosphate uptake rates as well as their morphological differences in P deplete versus P rich conditions. Our in vitro experiments showed that both Leptocylindrus species morphologically reacted similarly to phosphorus depletion and showed significantly elongated pervalvar axis in P depleted conditions if compared to P rich conditions. Also average chain lengths increased when in P depleted conditions. Two previously mentioned adaptations indicate their tendency to increase cellular surface areas available for alkaline phosphatase. Chlorophyll fluorescence of both species significantly decreased in P depleted medium. Although both species morphologically reacted similarly, our experiment demonstrated significant differences in physiological reactions to P depleted conditions.

The effects of two sized polystyrene nanoplastics on the growth, physiological functions, and toxin production of Alexandrium tamarense

Micro- and nano-plastics (MNPs) are increasingly prevalent pollutants in marine ecosystems and result in various deleterious effects on marine organisms. There have been studies evaluated the toxic effects of MNPs on marine microalgae, but few of them focused on the effects of MNPs on dinoflagellate species and their toxins production, which could have significant implications on human health and ecological safety in coastal areas. In this study, the common harmful algal blooms-causing dinoflagellate Alexandrium tamarense was exposed to 0.1 and 1 μm sized polystyrene nanoplastics (NPs) to investigate the responding patterns of population growth, multiple physiological functions, as well as the intracellular paralytic shellfish toxins (PSTs) productions. The results indicated the population growth, photosynthetic parameters, nutrients (nitrate and phosphate) uptake rates and extracellular carbonic anhydrase activities (CAext) were all inhibited by the two sized NPs, accompanied by the prolonged and more aggregated microalgal cells under the observation of scanning electron microscope (SEM), and the inhibition effects were more severe under 1 μm sized NPs than 0.1 μm sized NPs. Finally, we found the intracellular PSTs contents increased 73.59% exposed to 0.1 μm sized NPs while decreased 85.50% exposed to 1 μm sized NPs comparing the controls at 96 h, without significant changes of relative compositions. These results provided evidence that MNPs were toxic to A. tamarense and affected their intracellular PSTs productions within 96 h, which is critical to consider when evaluating the potential risks of MNPs in marine ecosystems.

Biodegradation of natural and synthetic endocrine-disrupting chemicals by aerobic granular sludge reactor: Evaluating estrogenic activity and estrogens fate.

In this study, the biodegradation of endocrine-disrupting chemicals (EDCs) (namely the natural and synthetic estrogens 17β-estradiol (E2) and 17α-ethinylestradiol (EE2), respectively) was assessed in an aerobic granular sludge (AGS) sequencing batch reactor (SBR) treating simulated domestic sewage. To better understand the fate of these compounds, their concentrations were determined in both liquid and solid (biomass) samples. Throughout the operation of the reactor, subjected to alternating anaerobic and aerated conditions, the removal of the hormones, both present in the influent at a concentration of 20μgL-1, amounted to 99% (for E2) and 93% (for EE2), with the latter showing higher resistance to biodegradation. Through yeast estrogen screen assays, an average moderate residual estrogenic activity (0.09μgL-1 EQ-E2) was found in the samples analysed. E2 and EE2 profiles over the SBR cycle suggest a rapid initial adsorption of these compounds on the granular biomass occurring anaerobically, followed by biodegradation under aeration. A possible sequence of steps for the removal of the micropollutants, including the key microbial players, was proposed. Besides the good capability of the AGS on EDCs removal, the results revealed high removal efficiencies (>90%) of COD, ammonium and phosphate. Most of the incoming organics (>80%) were consumed under anaerobic conditions, when phosphate was released (75.2 mgP L-1). Nitrification and phosphate uptake took place along the aeration phase, with effluent ammonium and phosphate levels around 2mgL-1. Although nitrite accumulation took place over the cycle, nitrate consisted of the main oxidized nitrogen form in the effluent. The specific ammonium and phosphate uptake rates attained in the SBR were found to be 3.3 mgNH4+-N gVSS-1.h-1 and 6.7 mgPO43--P gVSS-1 h-1, respectively, while the specific denitrification rate corresponded to 1.0 mgNOx--N gVSS-1 h-1.

The phytoremediation of water with high concentrations of nitrogen and phosphorus contamination by three selected wetland plants

High concentrations of nitrogen (N) (approximately 400 mg L −1) and phosphorus (P) (approximately 50 mg L−1) in wastewater can directly decrease the nutrient uptake rate of aquatic plants. Three floating wetland plants Myriophyllum elatinoides G., Eichhornia crassipes (Mart.) S., and Alternanthera philoxeroides (Mart.) G. were cultivated under different concentrations of N and P in microcosms to measure their potential capacity to absorb N and P. The results showed that M. elatinoides had the highest ammonia nitrogen (NH4+) and phosphate (PO43−) uptake rates at NH4+ and PO43− concentrations of 500 mg L−1 and 80 mg L−1, respectively, while E. crassipes had the highest nitrate (NO3−) uptake rate at NO3- concentrations of 50 mg L−1. These results, indicate that M. elatinoides absorbs NH4+ and PO43− more efficiently than E. crassipes and A. philoxeroides. The N/P ratio in wastewater impacted the removal of N and P by aquatic plants. The optimal N/P ratios for N and P removal by M. elatinoides, E. crassipes, and A. philoxeroides were 10:1, 20:1, and 20:1, respectively, with M. elatinoides having the highest N and P removal efficiency at this N/P ratio in the first 6 days of the experiment. The results indicated that M. elatinoides had a high potential for removal of N and P from wastewater with high concentrations of N and P, especially at the N/P ratio of 10:1 in the study. These results should provide direction in selection of plants for constructed wetland treatment systems of high concentration N and P wastewater.

Effect of sludge age on aerobic granular sludge: Addressing nutrient removal performance and biomass stability

The effect of the sludge retention time (SRT) on the stability and performance of an aerobic granular sludge (AGS) sequencing-batch reactor (SBR) on the simultaneous organic matter, nitrogen and phosphorus removal from simulated domestic wastewater was assessed in a long-term study (392 days). Operated under alternating anaerobic-aerobic conditions, the reactor was subjected to four experimental conditions: uncontrolled SRT (run I), SRT of 30 (run II), 20 (run III) and 15 days (run IV). The results showed that COD removal was kept stable and over 90 % throughout the SBR operation, regardless of the SRT. On the other hand, by allowing the sludge age to be dependent on natural solids washout by effluent withdrawal (SRT of 47–61 days), phosphate removal was substantially low (15 %), as also observed at an SRT of 30 days. Filamentous bacteria overgrowth was noticed at the later conditions, which affected the stability of the granular biomass, leading to a deterioration of its settling properties. The ratio between the sludge volume index after 30 and 5 min of settling (SVI30/SVI5) was around 0.70. Biological phosphate removal started to thrive at the sludge age of 20 days (35 %), reaching around 100 % at SRT of 15 days, at which the highest P-release/COD uptake ratio (0.14 mg P/mgCOD) and specific phosphate uptake (11.4 mgPO43−-P/(gVSS h)) were observed. Under the lowest SRT applied, the granules exhibited better structural and settling properties, with the SVI30/SVI5 ratio reaching almost 1.0. Moreover, nitrification was kept stable, and, even though nitrite build-up occurred during the SBR cycle, nitrate was the main oxidized nitrogen form in the effluent. Average COD, ammonium and total nitrogen removal amounted to 93 %, 97 % and 58 %. Cycle tests under normal and special conditions were carried out to assess specific nitrification, denitrification and phosphate uptake rates, and elucidate the key players in the biological conversions processes taking place in the AGS reactor. No phosphate uptake coupled to nitrate reduction was noticed, implying that P removal was not driven by denitrifying dephosphatation activity. To track the dynamics of important microbial functional groups, fluorescence in situ hybridization analysis was conducted.

Genotype difference in the physiological characteristics of phosphorus acquisition by wheat seedlings in alkaline soils

Phosphorus (P) in soils occurs predominately as insoluble inorganic P and organic P. However, key factors controlling P acquisition by wheat (Triticum aestivum L.) seedlings are unclear. In this study, the difference in the physiological characteristics of P acquisition in alkaline soils was investigated in wheat seedlings of two cultivars Aikang 58 and Zhoumai 22. The results indicated that the shoot P concentration of Aikang 58 was significantly higher than that of Zhoumai 22 when supplied with 0 and 70 kg/ha of pure P under field conditions. When cultured in sterile nutrition solutions with equimolar amounts of P corresponding to KH<sub>2</sub>PO<sub>4</sub>, Ca<sub>3</sub>(PO<sub>4</sub>)<sub>2</sub>, and Ca(H<sub>2</sub>PO<sub>4</sub>)<sub>2</sub> for 6 days, the P concentration in the shoots and roots of the seedlings of Aikang 58 was significantly higher than that of Zhoumai 22. However, the P concentration of seedlings of Aikang 58 did not exhibit a significant difference than that of Zhoumai 22 when cultured in phytic acid solution. Further studies suggested that the proton secretion rate was higher, and the root phosphatase activity was significantly lower in Aikang 58 compared with those in Zhoumai 22. After 48 h of successive P starvation, the inorganic phosphate (P<sub>i</sub>) uptake rate of Aikang 58 was significantly higher compared with that of Zhoumai 22. However, no significant differences existed in the root morphology between the two cultivars. Hence, the higher P acquisition in the wheat seedlings of Aikang 58 was attributed to a higher rate of proton secretion and a stronger capacity for P<sub>i</sub> uptake.

Evaluating the effect of air flow rate on hybrid and conventional membrane bioreactors: Implications on performance, microbial activity and membrane fouling

This study addressed the impact of air flow rate on the performance, membrane fouling behaviour and microbial community of a sequencing batch conventional membrane bioreactor (SB-MBR) and a sequencing batch hybrid membrane bioreactor (SB-HMBR) with carrier media for biofilm growth. Two different scenarios were evaluated: high (6.4 L min−1) and low (1.6 L min−1) air flow rates, associated with high (4.5 mg L−1) and low (1.5 mg L−1) dissolved oxygen (DO) concentrations and specific aeration demand per membrane area (SADm) of 0.426 and 0.106 m3 m−2 h−1, respectively. Both reactors were subjected to alternating non-aerated and aerated conditions for organic matter (as chemical oxygen demand - COD), nitrogen and phosphate removal from a municipal wastewater. From the bacterial community analysis, the key players in nutrient removal processes were assessed. The results showed that COD removal efficiencies were above 95% in both MBRs, regardless of the aeration intensity, while complete ammonium removal was observed at the higher DO. However, nitrifying activity was adversely affected under low DO levels. High nitrification levels were re-established faster in the hybrid MBR, thanks to the presence of biofilm, where nitrifying activity was favoured and the bacterial community profile did not exhibit substantial changes upon DO reduction. A higher denitrification potential was found for the carrier-based MBR, resulting in lower effluent nitrate concentrations. Regarding phosphorus removal, a slight improvement was observed in the SB-HMBR at reduced DO, while in the SB-MBR it remained practically constant. Moreover, the specific phosphate uptake rate exhibited a significant increase, especially in the hybrid MBR, reaching 44.6 mgP gVSS−1 h−1. At lower aeration rate, however, worse filterability and higher membrane fouling rates were observed, especially in the conventional MBR. Overall, the results demonstrated that the hybrid MBR better withstood the reduced air flow rate and DO as compared to the conventional counterpart.

Impact of aerobic availability of readily biodegradable COD on morphological stability of aerobic granular sludge

Operational disturbances in aerobic granular sludge (AGS) systems can result in aerobic availability of readily biodegradable COD (rbCOD). Different from activated sludge, morphological consequences on the short and long term are not well described in literature. This study investigated the effect of incomplete anaerobic uptake of acetate on the morphological and process stability of AGS using a lab-scale reactor. A fraction of the total acetate load was dosed aerobically, which was increased stepwise while monitoring granular morphology. A good granular morphology and an SVI of 40 ml/g were obtained during initial enrichment and maintained for ≤20% aerobic acetate load dosed at 4 mg COD/g VSS/h. Biological phosphorus removal efficiency was initially unaffected, but the aerobic acetate dosage rate did decrease the aerobic phosphate uptake rate. This led to loss of phosphorus removal for >20% aerobic acetate load dosed at 8 mg COD/g VSS/h over the course of 12 days. Subsequently, significant outgrowth formed on the granular surfaces and developed over time into finger-like structures. Under these high aerobic acetate loads the SVI increased to 80 ml/g and resulted in significant biomass washout due to deteriorating settling properties of the sludge. The sludge settleability and biological phosphorus removal recovered 10 days after aerobic feeding of acetate was stopped. Aerobic presence of rbCOD can be tolerated if mostly anaerobic acetate uptake is maintained, thereby ensuring stable granular morphology and good settleability. The high enrichment of phosphate accumulating organisms in the granular sludge through bottom-feeding and selective wasting of flocs makes aerobic granular sludge resilient to morphological deterioration in aerobic presence of rbCOD.

High Nitrogen and Phosphorous Acquisition by Belowground Parts of Caulerpa prolifera (Chlorophyta) Contribute to the Species' Rapid Spread in Ria Formosa Lagoon, Southern Portugal.

Despite worldwide proliferation of the genus Caulerpa and subsequent effects on benthic communities, little is known about the nutritional physiology of the Caulerpales. Here, we investigated the uptake rates of ammonium, nitrate, amino acids,and phosphate through the fronds and rhizoids+stolon, the internal translocation of nitrogen, and developed a nitrogen budget for the rapidly spreading Caulerpa prolifera in Ria Formosa lagoon, southern Portugal. Caulerpaprolifera acquired nutrients by both aboveground and belowground parts at similar rates, except nitrate, for which fronds showed 2-fold higher uptake rates. Ammonium was the preferential nitrogen source (81% of the total nitrogen acquisition), and amino acids, which accounted for a significant fraction of total N acquisition (19%), were taken up at faster rates than nitrate. Basipetal translocation of 15 N incorporated as ammonium was nearly 3-fold higher than acropetal translocation, whereas 15 N translocation as nitrate and amino acids was smaller but equal in either direction. The estimated total nitrogen acquisition by C.prolifera was 689 μmol · m-2 ·h-1 , whereas the total nitrogen requirement for growth was 672μmol·m-2 ·h-1 . The uptake of ammonium and amino acids by belowground parts accounted for the larger fraction of the total nitrogen acquisition of C.prolifera and is sufficient to satisfy the species nitrogen requirements for growth. This may be one reason explaining the fast spreading of the seaweed in the bare sediments of Ria Formosa where it does not have any macrophyte competitors and the concentration of nutrients is high.