Microbially Induced Calcium Carbonate Precipitation by Sporosarcina pasteurii: a Case Study in Optimizing Biological CaCO3 Precipitation.

Current production of traditional concrete requires enormous energy investment that accounts for approximately 5 to 8% of the world's annual CO2 production. Biocement is a building material that is already in industrial use and has the potential to rival traditional concrete as a more convenient and more environmentally friendly alternative. Biocement relies on biological structures (enzymes, cells, and/or cellular superstructures) to mineralize and bind particles in aggregate materials (e.g., sand and soil particles). Sporosarcina pasteurii is a workhorse organism for biocementation, but most research to date has focused on S. pasteurii as a building material rather than a biological system. In this review, we synthesize available materials science, microbiology, biochemistry, and cell biology evidence regarding biological CaCO3 precipitation and the role of microbes in microbially induced calcium carbonate precipitation (MICP) with a focus on S. pasteurii. Based on the available information, we provide a model that describes the molecular and cellular processes involved in converting feedstock material (urea and Ca2+) into cement. The model provides a foundational framework that we use to highlight particular targets for researchers as they proceed into optimizing the biology of MICP for biocement production.

Impaired cell growth under ammonium stress explained by modeling the energy cost of vacuole expansion in tomato leaves.

Ammonium (NH4 + )-based fertilization efficiently mitigates the adverse effects of nitrogen fertilization on the environment. However, high concentrations of soil NH4 + provoke growth inhibition, partly caused by the reduction of cell enlargement and associated with modifications of cell composition, such as an increase of sugars and a decrease in organic acids. Cell expansion depends largely on the osmotic-driven enlargement of the vacuole. However, the involvement of subcellular compartmentation in the adaptation of plants to ammonium nutrition has received little attention, until now. To investigate this, tomato (Solanum lycopersicum) plants were cultivated under nitrate and ammonium nutrition and the fourth leaf was harvested at seven developmental stages. The vacuolar expansion was monitored and metabolites and inorganic ion contents, together with intracellular pH, were determined. A data-constrained model was constructed to estimate subcellular concentrations of major metabolites and ions. It was first validated at the three latter developmental stages by comparison with subcellular concentrations obtained experimentally using non-aqueous fractionation. Then, the model was used to estimate the subcellular concentrations at the seven developmental stages and the net vacuolar uptake of solutes along the developmental series. Our results showed ammonium nutrition provokes an acidification of the vacuole and a reduction in the flux of solutes into the vacuoles. Overall, analysis of the subcellular compartmentation reveals a mechanism behind leaf growth inhibition under ammonium stress linked to the higher energy cost of vacuole expansion, as a result of alterations in pH, the inhibition of glycolysis routes and the depletion of organic acids.

Does energy cost constitute the primary cause of ammonium toxicity in plants?

Nitrate (NO3-) and ammonium (NH4+) are the main nitrogen (N) sources and key determinants for plant growth and development. In recent decades, NH4+, which is a double-sided N compound, has attracted considerable amounts of attention from researchers. Elucidating the mechanisms of NH4+ toxicity and exploring the means to overcome this toxicity are necessary to improve agricultural sustainability. In this review, we discuss the current knowledge concerning the energy consumption and production underlying NH4+ metabolism and toxicity in plants, such as N uptake; assimilation; cellular pH homeostasis; and functions of the plasma membrane (PM), vacuolar H+-ATPase and H+-pyrophosphatase (H+-PPase). We also discuss whether the overconsumption of energy is the primary cause of NH4+ toxicity or constitutes a fundamental strategy for plants to adapt to high-NH4+ stress. In addition, the effects of regulators on energy production and consumption and other physiological processes are listed for evaluating the possibility of high energy costs associated with NH4+ toxicity. This review is helpful for exploring the tolerance mechanisms and for developing NH4+-tolerant varieties as well as agronomic techniques to alleviate the effects of NH4+ stress in the field.

Assessment of anammox, microalgae and white-rot fungi-based processes for the treatment of textile wastewater.

The treatability of seven wastewater samples generated by a textile digital printing industry was evaluated by employing 1) anammox-based processes for nitrogen removal 2) microalgae (Chlorella vulgaris) for nutrient uptake and biomass production 3) white-rot fungi (Pleurotus ostreatus and Phanerochaete chrysosporium) for decolorization and laccase activity. The biodegradative potential of each type of organism was determined in batch tests and correlated with the main characteristics of the textile wastewaters through statistical analyses. The maximum specific anammox activity ranged between 0.1 and 0.2 g N g VSS-1 d-1 depending on the sample of wastewater; the photosynthetic efficiency of the microalgae decreased up to 50% during the first 24 hours of contact with the textile wastewaters, but it improved from then on; Pleurotus ostreatus synthetized laccases and removed between 20-62% of the colour after 14 days, while the enzymatic activity of Phanerochaete chrysosporium was inhibited. Overall, the findings suggest that all microbes have great potential for the treatment and valorisation of textile wastewater after tailored adaptation phases. Yet, the depurative efficiency can be probably enhanced by combining the different processes in sequence.

A metatranscriptomics-based assessment of small-scale mixing of sulfidic and oxic waters on redoxcline prokaryotic communities.

Lateral intrusions of oxygen caused by small-scale mixing are thought to shape microbial activity in marine redoxclines. To examine the response of prokaryotes to such mixing events we employed a shipboard mixing experiment in the euxinic central Baltic Sea: oxic, nitrate containing and sulfidic water samples without detectable oxygenized substances were incubated directly or after mixing. While nitrate, nitrite and ammonium concentrations stayed approximately constant in all incubations, we observed a decrease of sulfide after the contact with oxygen in the sulfide containing incubations. The transcription of marker genes from chemolithoauthotrophic key players including archaeal nitrifiers as well as gammaproteobacterial and campylobacterial autotrophic organisms that couple denitrification with sulfur-oxidation were followed at four time points within 8.5 h. The temporally contrasting transcriptional profiles of gammaproteobacterial and campylobacterial denitrifiers that depend on the same inorganic substrates pointed to a niche separation. Particular archaeal and campylobacterial marker genes involved in nitrification, denitrification and sulfur oxidation, which depend on oxidized substrates, were highly upregulated in the anaerobic sulfidic samples. We suggest that, despite the absence of measurable oxygenated compounds in the sulfidic water, frequent intermittent small-scale intrusions stimulate the permanent upregulation of genes involved in nitrification, denitrification and sulfur oxidation.

Modulating plant growth-metabolism coordination for sustainable agriculture.

Enhancing global food security by increasing the productivity of green revolution varieties of cereals risks increasing the collateral environmental damage produced by inorganic nitrogen fertilizers. Improvements in the efficiency of nitrogen use of crops are therefore essential; however, they require an in-depth understanding of the co-regulatory mechanisms that integrate growth, nitrogen assimilation and carbon fixation. Here we show that the balanced opposing activities and physical interactions of the rice GROWTH-REGULATING FACTOR 4 (GRF4) transcription factor and the growth inhibitor DELLA confer homeostatic co-regulation of growth and the metabolism of carbon and nitrogen. GRF4 promotes and integrates nitrogen assimilation, carbon fixation and growth, whereas DELLA inhibits these processes. As a consequence, the accumulation of DELLA that is characteristic of green revolution varieties confers not only yield-enhancing dwarfism, but also reduces the efficiency of nitrogen use. However, the nitrogen-use efficiency of green revolution varieties and grain yield are increased by tipping the GRF4-DELLA balance towards increased GRF4 abundance. Modulation of plant growth and metabolic co-regulation thus enables novel breeding strategies for future sustainable food security and a new green revolution.

Assessment of molecular detection of anaerobic ammonium-oxidizing (anammox) bacteria in different environmental samples using PCR primers based on 16S rRNA and functional genes.

Eleven published PCR primer sets for detecting genes encoding 16S ribosomal RNA (rRNA), hydrazine oxidoreductase (HZO), cytochrome cd 1-containing nitrite reductase (NirS), and hydrazine synthase subunit A (HzsA) of anaerobic ammonium-oxidizing (anammox) bacteria were assessed for the diversity and abundance of anammox bacteria in samples of three environments: wastewater treatment plant (WWTP), wetland of Mai Po Nature Reserve (MP), and the South China Sea (SCS). Consistent phylogenetic results of three biomarkers (16S rRNA, hzo, and hzsA) of anammox bacteria were obtained from all samples. WWTP had the lowest diversity with Candidatus Kuenenia dominating while the SCS was dominated by Candidatus Scalindua. MP showed the highest diversity of anammox bacteria including C. Scalindua, C. Kuenenia, and Candidatus Brocadia. Comparing different primer sets, no significant differences in specificity for 16S rRNA gene could be distinguished. Primer set CL1 showed relatively high efficiency in detecting the anammox bacterium hzo gene from all samples, while CL2 showed greater selectivity for WWTP samples. The recently reported primer sets of the hzsA gene resulted in high efficiencies in detecting anammox bacteria while nirS primer sets were more selective for specific samples. Results collectively indicate that the distribution of anammox bacteria is niche-specific within different ecosystems and primer specificity may cause biases on the diversity detected.

Engineering application of anaerobic ammonium oxidation process in wastewater treatment.

Anaerobic ammonium oxidation (Anammox), a promising biological nitrogen removal process, has been verified as an efficient, sustainable and cost-effective alternative to conventional nitrification and denitrification processes. To date, more than 110 full-scale anammox plants have been installed and are in operation, treating industrial NH4+-rich wastewater worldwide, and anammox-based technologies are flourishing. This review the current state of the art for engineering applications of the anammox process, including various anammox-based technologies, reactor selection and attempts to apply it at different wastewater plants. Process control and implementation for stable performance are discussed as well as some remaining issues concerning engineering application are exposed, including the start-up period, process disturbances, greenhouse gas emissions and especially mainstream anammox applications. Finally, further development of the anammox engineering application is proposed in this review.

Feasibility of two low-cost organic substrates for inducing denitrification in artificial recharge ponds: Batch and flow-through experiments.

Anaerobic batch and flow-through experiments were performed to assess the capacity of two organic substrates to promote denitrification of nitrate-contaminated groundwater within managed artificial recharge systems (MAR) in arid or semi-arid regions. Denitrification in MAR systems can be achieved through artificial recharge ponds coupled with a permeable reactive barrier in the form of a reactive organic layer. In arid or semi-arid regions, short-term efficient organic substrates are required due to the short recharge periods. We examined the effectiveness of two low-cost, easily available and easily handled organic substrates, commercial plant-based compost and crushed palm tree leaves, to determine the feasibility of using them in these systems. Chemical and multi-isotopic monitoring (δ15NNO3, δ18ONO3, δ34SSO4, δ18OSO4) of the laboratory experiments confirmed that both organic substrates induced denitrification. Complete nitrate removal was achieved in all the experiments with a slight transient nitrite accumulation. In the flow-through experiments, ammonium release was observed at the beginning of both experiments and lasted longer for the experiment with palm tree leaves. Isotopic characterisation of the released ammonium suggested ammonium leaching from both organic substrates at the beginning of the experiments and pointed to ammonium production by DNRA for the palm tree leaves experiment, which would only account for a maximum of 15% of the nitrate attenuation. Sulphate reduction was achieved in both column experiments. The amount of organic carbon consumed during denitrification and sulphate reduction was 0.8‰ of the total organic carbon present in commercial compost and 4.4% for the palm tree leaves. The N and O isotopic fractionation values obtained (εN and εO) were -10.4‰ and -9.0‰ for the commercial compost (combining data from both batch and column experiments), and -9.9‰ and -8.6‰ for the palm tree column, respectively. Both materials showed a satisfactory capacity for denitrification, but the palm tree leaves gave a higher denitrification rate and yield (amount of nitrate consumed per amount of available C) than commercial compost.

Assessment of municipal wastewaters at various stages of treatment process as potential growth media for Chlorella sorokiniana under different modes of cultivation.

Wastewater utilization for microalgal biomass production is potentially the most economical route for its fuel and feed applications. In this study, suitability of various wastewater streams within a domestic wastewater treatment plant was evaluated for microalgal cultivation. Pre-treatment methods were evaluated to minimize bacterial load. Biomass, cell physiology, nutrient removal efficiencies and biochemical constituents of Chlorella sorokiniana were investigated in influent (INF) and anaerobic tank centrate (AC) under mixotrophic (Mixo) and heterotrophic (Hetero) cultivation. Promising biomass (77.14mgL-1d-1), lipid (24.91mgL-1d-1), protein (22.36mgL-1d-1) and carbohydrate (20.10mgL-1d-1) productivities were observed in Mixo AC with efficient ammonium (94.29%) and phosphate (83.30%) removal. Supplementation of urea at a concentration of 1500mgL-1 further enhanced biomass (162.50mgL-1d-1), lipid (24.91mgL-1d-1), protein (22.36mgL-1d-1) and carbohydrate (20.10mgL-1d-1) productivities in Mixo AC. Urea supplemented mixotrophic cultivation of microalgae in AC is developed as a biomass production strategy.

Assessment of Heterotrophic Growth Supported by Soluble Microbial Products in Anammox Biofilm using Multidimensional Modeling.

Anaerobic ammonium oxidation (anammox) is known to autotrophically convert ammonium to dinitrogen gas with nitrite as the electron acceptor, but little is known about their released microbial products and how these are relative to heterotrophic growth in anammox system. In this work, we applied a mathematical model to assess the heterotrophic growth supported by three key microbial products produced by bacteria in anammox biofilm (utilization associated products (UAP), biomass associated products (BAP), and decay released substrate). Both One-dimensional and two-dimensional numerical biofilm models were developed to describe the development of anammox biofilm as a function of the multiple bacteria-substrate interactions. Model simulations show that UAP of anammox is the main organic carbon source for heterotrophs. Heterotrophs are mainly dominant at the surface of the anammox biofilm with small fraction inside the biofilm. 1-D model is sufficient to describe the main substrate concentrations/fluxes within the anammox biofilm, while the 2-D model can give a more detailed biomass distribution. The heterotrophic growth on UAP is mainly present at the outside of anammox biofilm, their growth on BAP (HetB) are present throughout the biofilm, while the growth on decay released substrate (HetD) is mainly located in the inner layers of the biofilm.

Comparative assessment of raw and digested pig slurry treatment in bioelectrochemical systems.

Both raw and anaerobically digested pig slurries were investigated in batch assays in two chambered bioelectrochemical systems (BES) run in Microbial Fuel Cell (MFC) and Microbial Electrolysis Cell (MEC) mode. Chemical Oxygen Demand (COD) removal, nitrogen recovery, cation transport and anode microbial population evolutions were assessed. The Anaerobic Digestion-MEC (AD-MEC) integrated system achieved the highest COD removal (60% in 48h); while the maximum NH4(+) removal efficiency (40%, with an ammonia flux of 8.86g N-NH4(+) d(-1)m(-2)) was achieved in MFC mode fed with digested pig slurry in 24h. On the other hand, the high pH (12.1) achieved in MEC mode (NaCl solution as catholyte), could favour ammonium recovery in a subsequent stripping and absorption process. Ammonia was the main cation involved in maintaining the electroneutrality between both compartments. Regarding microbial population, Desulfuromonadaceae, a known family of exoelectrogenic bacteria, was enriched under MEC mode, whereas hydrogenotrophic and methylotrophic methanogen phylotypes belonging to Thermoplasmatales were also favoured against acetotrophic Methanosaetaceae. From these results, the integration of anaerobic digestion in BES seems to be an interesting alternative for the treatment of complex substrates, since a polished effluent can be obtained and ammonium can be simultaneously recovered for further reuse as fertilizer.

A robust and cost-effective integrated process for nitrogen and bio-refractory organics removal from landfill leachate via short-cut nitrification, anaerobic ammonium oxidation in tandem with electrochemical oxidation.

A cost-effective process, consisting of a denitrifying upflow anaerobic sludge blanket (UASB), an oxygen-limited anoxic/aerobic (A/O) process for short-cut nitrification, and an anaerobic reactor (ANR) for anaerobic ammonia oxidation (anammox), followed by an electrochemical oxidation process with a Ti-based SnO2-Sb2O5 anode, was developed to remove organics and nitrogen in a sewage diluted leachate. The final chemical oxygen demand (COD), ammonia nitrogen (NH4(+)-N) and total nitrogen (TN) of 70, 11.3 and 39 (all in mg/L), respectively, were obtained. TN removal in UASB, A/O and ANR were 24.6%, 49.6% and 16.1%, respectively. According to the water quality and molecular biology analysis, a high degree of anammox besides short-cut nitrification and denitrification occurred in A/O. Counting for 16.1% of TN removal in ANR, at least 43.2-49% of TN was removed via anammox. The anammox bacteria in A/O and ANR, were in respective titers of (2.5-5.9)×10(9) and 2.01×10(10)copy numbers/(gSS).

Model-Based Feasibility Assessment of Membrane Biofilm Reactor to Achieve Simultaneous Ammonium, Dissolved Methane, and Sulfide Removal from Anaerobic Digestion Liquor.

In this study, the membrane biofilm reactor (MBfR) is proposed to achieve simultaneous removal of ammonium, dissolved methane, and sulfide from main-stream and side-stream anaerobic digestion liquors. To avoid dissolved methane stripping, oxygen is introduced through gas-permeable membranes, which also from the substratum for the growth of a biofilm likely comprising ammonium oxidizing bacteria (AOB), anaerobic ammonium oxidation (Anammox) bacteria, denitrifying anaerobic methane oxidation (DAMO) microorganisms, aerobic methane oxidizing bacteria (MOB), and sulfur oxidizing bacteria (SOB). A mathematical model is developed and applied to assess the feasibility of such a system and the associated microbial community structure under different operational conditions. The simulation studies demonstrate the feasibility of achieving high-level (>97.0%), simultaneous removal of ammonium, dissolved methane, and sulfide in the MBfRs from both main-stream and side-stream anaerobic digestion liquors through adjusting the influent surface loading (or hydraulic retention time (HRT)) and the oxygen surface loading. The optimal HRT was found to be inversely proportional to the corresponding oxygen surface loading. Under the optimal operational conditions, AOB, DAMO bacteria, MOB, and SOB dominate the biofilm of the main-stream MBfR, while AOB, Anammox bacteria, DAMO bacteria, and SOB coexist in the side-stream MBfR to remove ammonium, dissolved methane, and sulfide simultaneously.

Aquatic hazard, bioaccumulation and screening risk assessment for ammonium 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)-propanoate.

The fluoropolymer manufacturing industry is moving to alternative polymerization processing aid technologies with more favorable toxicological and environmental profiles as part of a commitment to curtail the use of long-chain perfluoroalkyl acids (PFAAs). To facilitate the environmental product stewardship assessment and premanufacture notification (PMN) process for a candidate replacement chemical, we conducted acute and chronic aquatic toxicity tests to evaluate the toxicity of ammonium 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)-propanoate (C6HF11O3.H3N) or the acid form of the substance to the cladoceran, Daphnia magna, the green alga, Pseudokirchneriella subcapitata, and a number of freshwater fish species including the rainbow trout, Oncorhynchus mykiss, In addition, testing with the common carp, Cyprinus carpio, was conducted to determine the bioconcentration potential of the acid form of the compound. Based on the relevant criteria in current regulatory frameworks, the results of the aquatic toxicity and bioconcentration studies indicate the substance is of low concern for aquatic hazard and bioconcentration in aquatic organisms. Evaluation of environmental monitoring data in conjunction with the predicted no effect concentration (PNEC) based on the available data suggest low risk to aquatic organisms.

Anammox moving bed biofilm reactor pilot at the 26th Ward wastewater treatment plants in Brooklyn, New York: start-up, biofilm population diversity and performance optimization.

New York City Environmental Protection in conjunction with City College of New York assessed the application of the anammox process in the reject water treatment using a moving bed biofilm reactor (MBBR) located at the 26th Ward wastewater treatment plant, in Brooklyn, NY. The single-stage nitritation/anammox MBBR was seeded with activated sludge and consequently was enriched with its own 'homegrown' anammox bacteria (AMX). Objectives of this study included collection of additional process kinetic and operating data and assessment of the effect of nitrogen loading rates on process performance. The initial target total inorganic nitrogen removal of 70% was limited by the low alkalinity concentration available in the influent reject water. Higher removals were achieved after supplementing the alkalinity by adding sodium hydroxide. Throughout startup and process optimization, quantitative real-time polymerase chain reaction (qPCR) analyses were used for monitoring the relevant species enriched in the biofilm and in the suspension. Maximum nitrogen removal rate was achieved by stimulating the growth of a thick biofilm on the carriers, and controlling the concentration of dissolved oxygen in the bulk flow and the nitrogen loading rates per surface area; all three appear to have contributed in suppressing nitrite-oxidizing bacteria activity while enriching AMX density within the biofilm.

Optimal cultivation of simultaneous ammonium and phosphorus removal aerobic granular sludge in A/O/A sequencing batch reactor and the assessment of functional organisms.

In this study, sequencing batch reactor (SBR) with an anaerobic/aerobic/anoxic operating mode was used to culture granular sludge. Optimal adjustment of cycle duration was achieved by the direction ofpH, oxidation reduction potential and dissolved oxygen parameters. The results showed that the treating efficiency was significantly improved as the cycle was shortened from 450 to 360 min and further to 200 min. Nitrogen and phosphorus removal were nearly quantitative after 50 days operation and maintained stable to the end of the study period. The typical cycle tests revealed that simultaneous denitrification and phosphorus removal occurred when aerobic granules were gradually formed. The nitrite effect tests showed that less than 4.8 mg N/L of the nitrite could enhance superficial specific aerobic phosphate uptake rate (SAPUR) under aerobic condition, indicating that the traditional method to evaluate the capability of total phosphate-accumulating organisms (PAOs) was inaccurate. Additionally, a high level of nitrite was detrimental to PAOs. A novel method was developed to determine the activity of each kind of PAOs and other denitrifying organisms. The results showed that (1) nitrate, besides nitrite, could also enhance SAPUR and (2) aerobic granular sludge could perform denitrification even when phosphate was not supplied under anoxic condition, suggesting that other denitrifying organisms besides denitrifying phosphate-accumulating organisms also contributed to denitrification.

Nitrogen and chemical oxygen demand removal from septic tank wastewater in subsurface flow constructed wetlands: substrate (cation exchange capacity) effects.

The current article focuses on chemical oxygen demand (COD) and nitrogen (ammonium and nitrate) removal performance from synthetic human wastewater as affected by different substrate rocks having a range of porosities and cation exchange capacities (CECs). The aggregates included lava rock, lightweight expanded shale, meta-basalt (control), and zeolite. The first three had CECs of 1 to 4 mequiv/100 gm, whereas the zeolite CEC was much greater (-80 mequiv/100 gm). Synthetic wastewater was gravity fed to each constructed wetland system, resulting in a 4-day retention time. Effluent samples were collected, and COD and nitrogen species concentrations measured regularly during four time periods from November 2008 through June 2009. Chemical oxygen demand and nitrogen removal fractions were not significantly different between the field and laboratory constructed wetland systems when corrected for temperature. Similarly, overall COD and nitrogen removal fractions were practically the same for the aggregate substrates. The important difference between aggregate effects was the zeolite's ammonia removal process, which was primarily by adsorption. The resulting single-stage nitrogen removal process may be an alternative to nitrification and denitrification that may realize significant cost savings in practice.

Efficiency assessment and pH effect in removing nitrogen and phosphorus by algae-bacteria combined system of Chlorella vulgaris and Bacillus licheniformis.

To achieve better removal of NH4(+) and TP in wastewater, a new algae-bacteria combined system of Chlorella vulgaris and Bacillus licheniformis was investigated in a 6-d experiment. The results showed that 78% of NH4(+) could be removed in the combined system, while 29% in single algae system and only 1% in single bacteria system. Approximately 92% of TP was removed in the combined system, compared with 55% and 78% in single algae and bacteria system, respectively. B. licheniformis was proven to be a growth-promoting bacterium for C. vulgaris by comparing Chl a concentrations in the single and combined systems. In the removal process, pH of the combined system was observed to reduce significantly from 7.0 to 3.5. Whereas with pH regulated to 7.0, higher removal efficiencies of NH4(+) (86%) and TP (93%) were achieved along with the recovery of algal cells and the increase of Chl a. These results suggest that nutrients in wastewater can be removed efficiently by the algae-bacteria combined system and pH control is crucial in the process.

Ammonium concentration of spent medium provides a noninvasive assessment of embryonic developmental potential in IVF.

BACKGROUND: Currently, morphological criteria are used to select embryos for transfer in in vitro fertilization (IVF)-embryo transfer. However, the implantation rate is only about 30%, indicating a need for a more efficient method of selecting high-quality embryos. This study investigated the relationship between medium ammonium concentration and IVF implantation rates and evaluated the possibility of using ammonium concentration to provide an objective, noninvasive, and simple means of assessment of embryo viability. METHODS: On day 3 after fertilization, we sampled the spent medium bathing embryos obtained from patients undergoing IVF or intracytoplasmic sperm injection (ICSI) at the Reproductive Medical Center of Sun Yat-sen Memorial Hospital, between September 2010 and January 2012. The concentration of ammonium was determined using a dry chemistry system. RESULTS: The ammonium concentration increased significantly during the period of culture of all embryos, but the increase was significantly greater in the group that, subsequently, did not achieve pregnancy (P < .001). An receiver-operating characteristic (ROC) curve indicated that ammonium concentration was highly predictive of embryo implantation failure (area under ROC curve = 0.838). Failure to achieve implantation had a predictive sensitivity of 71% and specificity of 79.6%. The accuracy of prediction of successful or failed implantation was 75.4%. In both IVF and ICSI, the embryo implantation rate decreased significantly as the medium ammonium concentration increased (P < .05). CONCLUSION: Measurement of the ammonium concentration in the spent medium may provide a new research direction for exploring a simple, rapid, and low-cost method for reliable prediction of embryo implantation with high sensitivity and specificity.