Redox regulation of chloroplast metabolism.

Regulation of enzyme activity based on thiol-disulfide exchange is a regulatory mechanism in which the protein disulfide reductase activity of thioredoxins (TRXs) plays a central role. Plant chloroplasts are equipped with a complex set of up to 20 TRXs and TRX-like proteins, the activity of which is supported by reducing power provided by photosynthetically reduced ferredoxin (FDX) with the participation of a FDX-dependent TRX reductase (FTR). Therefore, the FDX-FTR-TRXs pathway allows the regulation of redox-sensitive chloroplast enzymes in response to light. In addition, chloroplasts contain an NADPH-dependent redox system, termed NTRC, which allows the use of NADPH in the redox network of these organelles. Genetic approaches using mutants of Arabidopsis (Arabidopsis thaliana) in combination with biochemical and physiological studies have shown that both redox systems, NTRC and FDX-FTR-TRXs, participate in fine-tuning chloroplast performance in response to changes in light intensity. Moreover, these studies revealed the participation of 2-Cys peroxiredoxin (2-Cys PRX), a thiol-dependent peroxidase, in the control of the reducing activity of chloroplast TRXs as well as in the rapid oxidation of stromal enzymes upon darkness. In this review, we provide an update on recent findings regarding the redox regulatory network of plant chloroplasts, focusing on the functional relationship of 2-Cys PRXs with NTRC and the FDX-FTR-TRXs redox systems for fine-tuning chloroplast performance in response to changes in light intensity and darkness. Finally, we consider redox regulation as an additional layer of control of the signaling function of the chloroplast.

Frailty Assessment in a Stable COPD Cohort: Is There a COPD-Frail Phenotype?

The frailty syndrome increases the morbidity/mortality in older adults, and several studies have shown a higher prevalence of this syndrome in patients with Chronic Obstructive Pulmonary Disease (COPD). The aim of this study was to identify the characteristics of frail patients with COPD to define a new phenotype called "COPD-frail." We conducted a cross-sectional study in a cohort of patients with stable COPD, classified as either frail, pre-frail, or non-frail. Sociodemographic, clinical, and biochemical variables were compared between the three groups of patients. The study included 127 patients, of which 31 were frail, 64 were pre-frail, and 32 non-frail. All subjects had FEV1/FVC below the lower limit of normal (range Z-score: -1.66 and -5.32). Patients in the frail group showed significantly higher scores in the mMRC (modified Medical Research Council) scale, the CAT (COPD Assessment Test), and the BODE (Body mass index, airflow Obstruction, Dyspnea, and Exercise capacity) index. They also showed differences in symptoms according to GOLD (Global Initiative for Chronic Obstructive Lung Disease), as well as more COPD exacerbations, less physical activity, more anxiety and depression symptoms based on HADS (Hospital Anxiety and Depression Scale), and lower hemoglobin, hematocrit, and 25-hydroxycholecalciferol levels. Variables with independent association with frailty included the mMRC score, the HAD index for depression and age. In summary, differential characteristics of frail patients with COPD encourage the definition of a "COPD-frail" phenotype that-if identified early-would allow performing interventions to prevent a negative impact on the morbidity/mortality of these patients.

Metabolic Control of Tobacco Pollination by Sugars and Invertases.

Pollination in flowering plants is initiated by germination of pollen grains on stigmas followed by fast growth of pollen tubes representing highly energy-consuming processes. The symplastic isolation of pollen grains and tubes requires import of Suc available in the apoplast. We show that the functional coupling of Suc cleavage by invertases and uptake of the released hexoses by monosaccharide transporters are critical for pollination in tobacco (Nicotiana tabacum). Transcript profiling, in situ hybridization, and immunolocalization of extracellular invertases and two monosaccharide transporters in vitro and in vivo support the functional coupling in supplying carbohydrates for pollen germination and tube growth evidenced by spatiotemporally coordinated expression. Detection of vacuolar invertases in maternal tissues by these approaches revealed metabolic cross talk between male and female tissues and supported the requirement for carbohydrate supply in transmitting tissue during pollination. Tissue-specific expression of an invertase inhibitor and addition of the chemical invertase inhibitor miglitol strongly reduced extracellular invertase activity and impaired pollen germination. Measurements of (competitive) uptake of labeled sugars identified two import pathways for exogenously available Suc into the germinating pollen operating in parallel: direct Suc uptake and via the hexoses after cleavage by extracellular invertase. Reduction of extracellular invertase activity in pollen decreases Suc uptake and severely compromises pollen germination. We further demonstrate that Glc as sole carbon source is sufficient for pollen germination, whereas Suc is supporting tube growth, revealing an important regulatory role of both the invertase substrate and products contributing to a potential metabolic and signaling-based multilayer regulation of pollination by carbohydrates.

The Functional Relationship between NADPH Thioredoxin Reductase C, 2-Cys Peroxiredoxins, and m-Type Thioredoxins in the Regulation of Calvin-Benson Cycle and Malate-Valve Enzymes in Arabidopsis.

The concerted regulation of chloroplast biosynthetic pathways and NADPH extrusion via malate valve depends on f and m thioredoxins (Trxs). The finding that decreased levels of the thiol-peroxidase 2-Cys peroxiredoxin (Prx) suppress the severe phenotype of Arabidopsis mutants lacking NADPH-dependent Trx reductase C (NTRC) and Trxs f uncovered the central function of the NTRC-2-Cys-Prx redox system in chloroplast performance. These results suggest that Trxs m are also regulated by this system; however, the functional relationship between NTRC, 2-Cys Prxs, and m-type Trxs is unknown. To address this issue, we generated Arabidopsis thaliana mutants combining deficiencies in NTRC, 2-Cys Prx B, Trxs m1, and m4. The single trxm1 and trxm4 mutants showed a wild-type phenotype, growth retardation being noticed only in the trxm1m4 double mutant. Moreover, the ntrc-trxm1m4 mutant displayed a more severe phenotype than the ntrc mutant, as shown by the impaired photosynthetic performance, altered chloroplast structure, and defective light-dependent reduction in the Calvin-Benson cycle and malate-valve enzymes. These effects were suppressed by the decreased contents of 2-Cys Prx, since the quadruple ntrc-trxm1m4-2cpb mutant displayed a wild-type-like phenotype. These results show that the activity of m-type Trxs in the light-dependent regulation of biosynthetic enzymes and malate valve is controlled by the NTRC-2-Cys-Prx system.

Energy and Nutrients from Apple Waste Using Anaerobic Digestion and Membrane Technology.

The worldwide increment of food waste requires innovative management solutions, aligned with sustainability, energy, and food security. Anaerobic digestion (AD), followed by nutrient recovery, may be considered an interesting approach. This study proposed a co-digestion of apple pomace (AP) with swine manure (SM) to study the effect of different proportions of AP (0, 7.5, 15, and 30%, on a volatile solids (VS) basis) on the methane production and the stability of the process. Subsequently, the gas-permeable membrane (GPM) technology was applied to recover nitrogen (N) as ammonium sulfate (bio-based fertilizer) from the digestates produced after the AD of 7.5% of AP and SM, and SM alone. The results showed that the co-digestion of 7.5% and 15% of AP with SM presented a methane production similar to the AD of SM alone (with 412.3 ± 62.6, 381.8 ± 134.1, and 421.7 ± 153.6 mL g VS-1 day-1, respectively). The later application of the GPM technology on the resulting digestates, with SM alone and with 7.5% of AP with SM, showed total ammoniacal N recovery rates of 33 and 25.8 g N m-2 d-1, respectively. Therefore, the AP valorization through the AD process, followed by N recovery from the digestate, could be a good management strategy.

Comparison of the Ammonia Trapping Performance of Different Gas-Permeable Tubular Membrane System Configurations.

The technology of gas-permeable tubular membranes (GPMs) is promising in reducing ammonia emissions from livestock manure, capturing NH3 in an acidic solution, and obtaining final products suitable for valorization as fertilizers, in line with the principles of the circular economy. This study aimed to evaluate the performance of several e-PTFE membrane systems with different configurations for the recovery of NH3 released from pig slurry. Ten different configurations were tested: only a submerged membrane, only a suspended membrane in the same chamber, only a suspended membrane in an annex chamber, a submerged membrane + a suspended membrane in the same chamber, and a submerged membrane + a suspended membrane in an annex chamber, considering in each case the scenarios without and with agitation and aeration of the slurry. In all tests, sulfuric acid (1N H2SO4) was used as the NH3 capture solution, which circulated at a flow rate of 2.1 L·h-1. The results showed that NH3-N removal rates ranged from 36-39% (for systems with a single submerged or suspended membrane without agitation or aeration of the slurry) to 70-72% for submerged + suspended GPM systems with agitation and aeration. In turn, NH3-N recovery rates were found to be between 44-54% (for systems with a single membrane suspended in an annex compartment) and 88-91% (for systems based on a single submerged membrane). However, when choosing a system for farm deployment, it is essential to consider not only the capture and recovery performance of the system, but also the investment and operating costs (ranging from 9.8 to 21.2 €/kg N recovered depending on the selected configuration). The overall assessment suggests that the simplest systems, based on a single membrane, may be the most recommendable.

Long-term preservation of anammox bacteria.

Deposit of useful microorganisms in culture collections requires long-term preservation and successful reactivation techniques. The goal of this study was to develop a simple preservation protocol for the long-term storage and reactivation of the anammox biomass. To achieve this, anammox biomass was frozen or lyophilized at two different freezing temperatures (-60°C and in liquid nitrogen (-200°C)) in skim milk media (with and without glycerol), and the reactivation of anammox activity was monitored after a 4-month storage period. Of the different preservation treatments tested, only anammox biomass preserved via freezing in liquid nitrogen followed by lyophilization in skim milk media without glycerol achieved stoichiometric ratios for the anammox reaction similar to the biomass in both the parent bioreactor and in the freshly harvested control treatment. A freezing temperature of -60°C alone, or in conjunction with lyophilization, resulted in the partial recovery of the anammox bacteria, with an equal mixture of anammox and nitrifying bacteria in the reactivated biomass. To our knowledge, this is the first report of the successful reactivation of anammox biomass preserved via sub-zero freezing and/or lyophilization. The simple preservation protocol developed from this study could be beneficial to accelerate the integration of anammox-based processes into current treatment systems through a highly efficient starting anammox biomass.

Effect of operational conditions on the degradation of organic matter and development of microalgae-bacteria consortia when treating swine slurry.

There is great controversy regarding the best substrate (fresh or anaerobically digested swine slurry) for the development of microalgae-bacteria consortia. This study aims to elucidate the best substrate by assessing biomass productivity, microorganism predominance, and their ability for organic matter removal. In addition to the different substrates, different operational conditions and influent strengths were evaluated. Increasing organic matter content when favourable temperature and illumination conditions were present improved biomass production. However, these conditions were not favourable for microalgal growth, but they were favourable for bacteria. Regardless of the operational conditions, reactors fed with fresh slurry not only resulted in the highest biomass productivity, but also the greatest removal of total and soluble chemical oxygen demand (COD). On the other hand, reactors fed with digested slurry showed biomass productivity and COD removal values lower than those obtained for reactors fed with fresh slurry, most probably due to the recalcitrant nature of the former. Nevertheless, digested slurry was the substrate more appropriate for microalgae growth under harsh operational conditions (16 °C and 9-h illumination) at low influent strength and optimum operational conditions (30 °C and 24-h illumination) at higher influent strength.

Current Knowledge on Mechanisms Preventing Photosynthesis Redox Imbalance in Plants.

Photosynthesis includes a set of redox reactions that are the source of reducing power and energy for the assimilation of inorganic carbon, nitrogen and sulphur, thus generating organic compounds, and oxygen, which supports life on Earth. As sessile organisms, plants have to face continuous changes in environmental conditions and need to adjust the photosynthetic electron transport to prevent the accumulation of damaging oxygen by-products. The balance between photosynthetic cyclic and linear electron flows allows for the maintenance of a proper NADPH/ATP ratio that is adapted to the plant's needs. In addition, different mechanisms to dissipate excess energy operate in plants to protect and optimise photosynthesis under adverse conditions. Recent reports show an important role of redox-based dithiol-disulphide interchanges, mediated both by classical and atypical chloroplast thioredoxins (TRXs), in the control of these photoprotective mechanisms. Moreover, membrane-anchored TRX-like proteins, such as HCF164, which transfer electrons from stromal TRXs to the thylakoid lumen, play a key role in the regulation of lumenal targets depending on the stromal redox poise. Interestingly, not all photoprotective players were reported to be under the control of TRXs. In this review, we discuss recent findings regarding the mechanisms that allow an appropriate electron flux to avoid the detrimental consequences of photosynthesis redox imbalances.

A novel approach for nutrients recovery from municipal waste as biofertilizers by combining electrodialytic and gas permeable membrane technologies.

The recovery of valuable materials from waste fits the principle of circular economy and sustainable use of resources, but contaminants in the waste are still a major obstacle. This works proposes a novel approach to recover high-purity phosphorus (P) and nitrogen (N) from digestate of municipal solid waste based on the combination of two independent membrane processes: electrodialytic (ED) process to extract P, and gas permeable membranes (GPM) for N extraction. A laboratory ED cell was adapted to accommodate a GPM. The length of waste compartment (10 cm; 15 cm), current intensity (50 mA; 75 mA) and operation time (9 days; 12 days) were the variables tested. 81% of P in the waste was successfully extracted to the anolyte when an electric current of 75 mA was applied for 9 days, and 74% of NH4+ was extracted into an acid-trapping solution. The two purified nutrient solutions were subsequently used in the synthesis of a biofertilizer (secondary struvite) through precipitation, achieving an efficiency of 99.5%. The properties of the secondary struvite synthesized using N and P recovered from the waste were similar to secondary struvite formed using synthetic chemicals but the costs were higher due to the need to neutralize the acid-trapping solution, highlighting the need to further tune the process and make it economically more competitive. The high recycling rates of P and N achieved are encouraging and widen the possibility of replacing synthetic fertilizers, manufactured from finite sources, by secondary biofertilizers produced using nutrients extracted from wastes.

Pilot-Scale Demonstration of Membrane-Based Nitrogen Recovery from Swine Manure.

Gas-permeable membranes technology presents a high potential for nitrogen (N) recovery from wastewaters rich in ammonia (NH3). The EU project Ammonia Trapping (AT) is aimed at transferring knowledge from the lab-scale level to on-farm pilot-scale level, using this technology to recover NH3 from livestock wastewaters. The goal of this study is to report the results of an on-farm pilot-scale demonstration plant using gas-permeable membranes to recover N from raw swine manure. After a setup optimization of the plant, stable, and continuous operation was achieved. The maximum NH3 recovery rate obtained was 38.20 g NH3-N m-2 membrane day-1. This recovery rate was greatly affected by the temperature of the process. In addition to its contribution to NH3 emissions reduction, this technology contributes to the recovery of nutrients in the form of a concentrated stable ammonium sulphate solution. This solution contained 3.2% of N, which makes it suitable for fertigation. The economic approach revealed an economic feasibility of the technology, resulting in a cost of 2.07 € per kg N recovered.

Lactose malabsorption and intolerance: a review.

Food lactose and lactose intolerance are today hot topics in the field of food and nutrition. About 70% of the adult world population is lactose-intolerant, due to low levels of intestinal lactase, also called lactase-phlorizin hydrolase (LPH), a ß-d-galactosidase found in the apical surface of the intestinal microvilli. This may be due to the loss of intestinal lactase in adulthood, a condition transmitted by an autosomal recessive gene, which differs in humans according to race. According to the cultural-historical hypothesis, the mutation that allows the metabolization of lactose appeared about 10 000 years ago in the inhabitants of Northern Europe where mammalian milk continued in the diet after weaning, and lactase-persistent populations were genetically selected in some areas. Many intolerant individuals can tolerate low levels of lactose in their daily diet. Probiotics have also been proposed as an alternative that could avoid some symptoms of lactose intolerance. Many products are marketed nowadays as alternatives to dairy products for lactose-intolerant individuals. However, the rules for low-lactose foods are currently not harmonised in the European Union. As scientific knowledge on lactose intolerance has notably advanced in recent decades, the aim of this work was to review the current state of the knowledge on lactose and lactose intolerance, its diagnosis and clinical management, and the various food products that are offered specifically for non-tolerant individuals.

An event of alternative splicing affects the expression of the NTRC gene, encoding NADPH-thioredoxin reductase C, in seed plants.

The NTRC gene encodes a NADPH-dependent thioredoxin reductase with a joint thioredoxin domain, exclusive of photosynthetic organisms. An updated search shows that although most species harbor a single copy of the NTRC gene, two copies were identified in different species of the genus Solanum, Glycine max and the moss Physcomitrella patens. The phylogenetic analysis of NTRCs from different sources produced a tree with the major groups of photosynthetic organisms: cyanobacteria, algae and land plants, indicating the evolutionary success of the NTRC gene among photosynthetic eukaryotes. An event of alternative splicing affecting the expression of the NTRC gene was identified, which is conserved in seed plants but not in algae, bryophytes and lycophytes. The alternative splicing event results in a transcript with premature stop codon, which would produce a truncated form of the enzyme. The standard splicing/alternative splicing (SS/AS) transcripts ratio was higher in photosynthetic tissues from Arabidopsis, Brachypodium and tomato, in line with the higher content of the NTRC polypeptide in these tissues. Moreover, environmental stresses such as cold or high salt affected the SS/AS ratio of the NTRC gene transcripts in Brachypodium seedlings. These results suggest that the alternative splicing of the NTRC gene might be an additional mechanism for modulating the content of NTRC in photosynthetic and non-photosynthetic tissues of seed plants.

Propidium Monoazide Coupled with PCR Predicts Infectivity of Enteric Viruses in Swine Manure and Biofertilized Soil.

The use of propidium monoazide (PMA) coupled with real-time PCR (RT-qPCR or qPCR for RNA or DNA viruses, respectively) was assessed to discriminate infectious enteric viruses in swine raw manure, swine effluent from anaerobic biodigester (AB) and biofertilized soils. Those samples were spiked either with infectious and heat-inactivated human adenovirus-2 (HAdV-2) or mengovirus (vMC0), and PMA-qPCR/RT-qPCR allowed discriminating inactivated viruses from the infective particles, with significant reductions (>99.9%). Then, the procedure was further assayed to evaluate the presence and stability of two non-cultivable viruses (porcine adenovirus and rotavirus A) in natural samples (swine raw manure, swine effluent from AB and biofertilized soils); it demonstrated viral inactivation during the storage period at 23 °C. As a result, the combination of PMA coupled to real-time PCR can be a promising alternative for prediction of viral infectivity in comparison to more labour-intensive and costly techniques such as animal or tissue-culture infectivity methods, and for those viruses that do not have currently available cell culture techniques.

Human recombinant anti-transglutaminase antibody testing is useful in the diagnosis of silent coeliac disease in a selected group of at-risk patients.

BACKGROUND: Functional dyspepsia, unexplained chronic hypertransaminasaemia (CHT) and hepatitis C virus (HCV) are common gastrointestinal situations that have been related to coeliac disease. Antibodies to tissue transglutaminase (tTG) have been claimed recently to be highly effective as a screening method for coeliac disease. AIM: To assess the prevalence of coeliac disease by means of detection of antibodies against human tTG in the above-mentioned groups of patients. PATIENTS AND METHODS: A control group consisted of 165 normal blood donors. Patient groups comprised 90 CHT patients, 102 HCV patients and 92 functional dyspepsia patients. All patients were tested for anti-tTG (immunoglobulin A, IgA) antibodies. Anti-endomysium (IgA) antibodies (AEA) and antigliadin (IgA) antibodies (AGA) and antigliadin (immunoglobulin G, IgG) antibodies (AGG) were also tested. When anti-tTG or AEA was positive, a duodenal biopsy was recommended. RESULTS: One of 165 blood donors, three of 92 functional dyspepsia patients, four of 90 CHT patients and none of 102 HCV patients were positive for anti-tTG antibodies. In the anti-tTG-positive group, all but one were AEA-positive. There were no AEA- or AGA IgA-positives that revealed a negative anti-tTG test. Duodenal biopsy confirmed a diagnosis of coeliac disease in all the cases. Statistically significant differences were found between the controls and the functional dyspepsia group and between the controls and the CHT group, but not between the controls and the HCV group. CONCLUSIONS: Both CHT and functional dyspepsia may represent a true oligosymptomatic form of coeliac disease. In such conditions, the detection of anti-tTG antibodies is useful as a screening method. Coeliac disease is not an autoimmune manifestation of HCV, so screening for coeliac disease in HCV patients cannot be recommended.

Optimized determination of calcium in grape juice, wines, and other alcoholic beverages by atomic absorption spectrometry.

This paper describes a study of the different methods of sample preparation for the determination of calcium in grape juice, wines, and other alcoholic beverages by flame atomic absorption spectrometry; results are also reported for the practical application of these methods to the analysis of commercial samples produced in Spain. The methods tested included dealcoholization, dry mineralization, and wet mineralization with heating by using different acids and/or mixtures of acids. The sensitivity, detection limit, accuracy, precision, and selectiviy of each method were established. Such research is necessary because of the better analytical indexes obtained after acid digestion of the sample, as recommended by the European Union, which advocates the direct method. In addition, although high-temperature mineralization with an HNO3-HCIO4 mixture gave the best analytical results, mineralization with nitric acid at 80 degrees C for 15 min gave the most satisfactory results in all cases, including those for wines with high levels of sugar and beverages with high alcoholic content. The results for table wines subjected to the latter treatment had an accuracy of 98.70-99.90%, a relative standard deviation of 2.46%, a detection limit of 19.0 microg/L, and a determination limit of 31.7 microg/L. The method was found to be sufficiently sensitive and selective. It was applied to the determination of Ca in grape juice, different types of wines, and beverages with high alcoholic content, all of which are produced and widely consumed in Spain. The values obtained for Ca were 90.00 +/- 20.40 mg/L in the grape juices, 82.30 +/- 23.80 mg/L in the white wines, 85.00 +/- 30.25 mg/L in the sweet wines, 84.92 +/- 23.11 mg/L in the red wines, 85.75 +/- 27.65 mg/L in the rosé wines, 9.51 +/- 6.65 mg/L in the brandies, 11.53 +/- 6.55 mg/L in the gin, 7.3 +/- 6.32 mg/L in the pacharán, and 8.41 +/- 4.85 mg/L in the anisettes. The method is therefore useful for routine analysis in the quality control of these beverages.

Fetal myocardial morphological and functional changes associated with well-controlled gestational diabetes.

OBJECTIVE: To make a global evaluation of the fetal myocardial changes in a well-controlled gestational diabetic population. STUDY DESIGN: Twenty-four pregnant well-controlled diabetic patients were selected. Sixteen normal pregnancies were randomly collected as a control group. Measurements of morphological and functional myocardial parameters were performed. Data from the left ventricular outflow tract and peripheral Doppler data were obtained. RESULT: The thickness of the interventricular septum was increased in diabetic pregnancies (p < 0.001). Tricuspid E/A index was the only functional parameter showing a significant variation, with lower values in diabetic pregnancies. Doppler parameters from the left ventricular outflow tract and peripheral Doppler waveforms were similar between groups. CONCLUSION: A tendency towards interventricular septum hypertrophy was observed even in well-controlled diabetic pregnancies. Mild hypertrophic cardiac changes were not associated with abnormal cardiac function or signs of left ventricular outflow obstruction, although minor changes in right ventricular diastolic function were recorded.

Extracellular invertase is involved in the regulation of clubroot disease in Arabidopsis thaliana.

Clubroot disease of Brassicaceae is caused by an obligate biotrophic protist, Plasmodiophora brassicae. During root gall development, a strong sink for assimilates is developed. Among other genes involved in sucrose and starch synthesis and degradation, the increased expression of invertases has been observed in a microarray experiment, and invertase and invertase inhibitor expression was confirmed using promoter::GUS lines of Arabidopsis thaliana. A functional approach demonstrates that invertases are important for gall development. Different transgenic lines expressing an invertase inhibitor under the control of two root-specific promoters, Pyk10 and CrypticT80, which results in the reduction of invertase activity, showed clearly reduced clubroot symptoms in root tissue with highest promoter expression, whereas hypocotyl galls developed normally. These results present the first evidence that invertases are important factors during gall development, most probably in supplying sugars to the pathogen. In addition, root-specific repression of invertase activity could be used as a tool to reduce clubroot symptoms.

Nitrogen transformations under different conditions in open ponds by means of microalgae-bacteria consortium treating pig slurry.

Four open ponds inoculated with microalgae-bacteria consortium treating different swine slurries (fresh and anaerobically digested) were evaluated in terms of nitrogen transformation under optimal and real conditions of temperature and illumination. Ammonium complete depletion was not achieved. Ponds operated under real conditions presented lower ammonium removal. Elimination capacities were around 26 mg N/Ld and were subsequently increased with increasing inlet ammonium loading rate. Different nitrogen transformation was observed depending on substrate source. When anaerobically digested slurry was fed to the ponds, nitrification followed by biomass uptake and denitrification were the main nitrogen transformation taking place depending on inlet ammonium loading rate and operational conditions. Ponds fed with fresh slurry exhibited denitrification as the main nitrogen removal mechanism for the pond operated under real conditions while under optimal conditions stripping, denitrification and biomass uptake contributed similarly. Therefore, this study confirmed that the so-claimed nitrogen recovery by microalgae biomass is frequently overestimated.