Nitrogen removal from wastewater by an immobilized consortium of microalgae-bacteria in hybrid hydrogels.

The high content of nitrogen in wastewater brings some operational, technical, and economical issues in conventional technologies. The aim of this study was to evaluate the nitrogen removal by hybrid hydrogels containing consortium microalgae-nitrifying bacteria in the presence of activated carbon (AC) used as an adsorbent of inhibitory substances. Hybrid hydrogels were synthesized from polyvinyl alcohol (PVA), sodium alginate (SA), biomass (microalgae-nitrifying bacteria), and AC. The hybrid hydrogels were evaluated based on the change in ammonium (NH4), nitrate (NO3), and chemical demand of oxygen (COD) concentrations, nitrification rate, and other parameters during 72 h. Results indicated that NH4 removal was more effective for hydrogels without AC than with AC, without significant differences regarding consortium biomass concentration (5 or 16%), presenting final concentrations of 3.13 and 3.75 mg NH4/L for hydrogels with 5 and 16% of the biomass, respectively. Regarding NO3 production, hydrogels without AC reached concentrations of 25.9 and 39.77 mg NO3/L for 5 and 16% of the biomass, respectively, while treatments with AC ended with 2.17 and 1.37 mg NO3/L. This confirms that hydrogels can carry out the nitrification process and do not need AC to remove potential inhibitors. The best performance was observed for the hydrogel with 5% of biomass without AC with a nitrification rate of 0.43 mg N/g TSS·h.

Influence of wastewater type in the effects caused by titanium dioxide nanoparticles in the removal of macronutrients by activated sludge.

The imminent arrival of nanoparticles (NPs) to the wastewater treatment plants (WWTP) brings concern about their effects, which can be related to the wastewater composition. In this work, the effects of titanium dioxide (TiO2) NPs in the removal of carbon, nitrogen, and phosphorus by activated sludge bioreactors during the treatment of synthetic, raw, and filtered wastewaters were evaluated. Floc size, compaction of sludge, and morphological interactions between sludge and NPs were also determined. The main effect of TiO2 NPs was the inhibition of up to 22% in the removal of ammonia nitrogen for all types of wastewaters. This effect is strong dependent on combined factors of TiO2 NPs concentration and content of organic matter and ammonia in wastewater. The removal of dissolved organic carbon was affected by TiO2 NPs in lower level (up to 6%) than nitrogen removal for all types of wastewaters. Conversely to adverse effects, the removals of orthophosphate in the presence of TiO2 NPs were improved by 34%, 16%, and 55% for synthetic, raw, and filtered wastewater, respectively. Compaction of the sludge was also enhanced as the concentrations of NPs increased without alterations in the floc size for all types of wastewaters. Based on TEM and STEM imaging, the main interaction between TiO2 NPs and the activated sludge flocs was the adsorption of NPs on cell membrane. This means that NPs can be attached to cell membrane during aerobic wastewater treatment, and potentially disrupt this membrane. The effects of TiO2 NPs on macronutrient removal clearly depended on wastewater characteristics; hence, the use of realistic media is highly encouraged for ecotoxicological experiments involving NPs.

Changes in bacterial diversity of activated sludge exposed to titanium dioxide nanoparticles.

The rapid growth of the use of nanomaterials in different modern industrial branches makes the study of the impact of nanoparticles on the human health and environment an urgent matter. For instance, it has been reported that titanium dioxide nanoparticles (TiO2 NPs) can be found in wastewater treatment plants. Previous studies have found contrasting effects of these nanoparticles over the activated sludge process, including negative effects on the oxygen uptake. The non-utilization of oxygen reflects that aerobic bacteria were inhibited or decayed. The aim of this work was to study how TiO2 NPs affect the bacterial diversity and metabolic processes on an activated sludge. First, respirometry assays of 8 h were carried out at different concentrations of TiO2 NPs (0.5-2.0 mg/mL) to measure the oxygen uptake by the activated sludge. The bacterial diversity of these assays was determined by sequencing the amplified V3-V4 region of the 16S rRNA gene using Illumina MiSeq. According to the respirometry assays, the aerobic processes were inhibited in a range from 18.5 ± 4.8% to 37.5 ± 2.0% for concentrations of 0.5-2.0 mg/mL TiO2 NPs. The oxygen uptake rate was affected mainly after 4.5 h for concentrations higher than 1.0 mg/mL of these nanoparticles. Results indicated that, in the presence of TiO2 NPs, the bacterial community of activated sludge was altered mainly in the genera related to nitrogen removal (nitrogen assimilation, nitrification and denitrification). The metabolic pathways prediction suggested that genes related to biofilm formation were more sensitive than genes directly related to nitrification-denitrification and N-assimilation processes. These results indicated that TiO2 NPs might modify the bacteria diversity in the activated sludge according to their concentration and time of exposition, which in turn impact in the performance of the wastewater treatment processes.

Cumulative effects of titanium dioxide nanoparticles in UASB process during wastewater treatment.

Titanium dioxide nanoparticles (TiO2 NPs) are widely used in consumer products and one of their major fate is the wastewater treatment plants. However, NPs eventually arrive to aquatic and terrestrial ecosystems via treated water and biosolids, respectively. Since low concentration of NPs is accumulating in the upflow anaerobic sludge blanket (UASB) reactors that treat wastewater and reclaim water quality, the accumulation of TiO2 NPs in these reactors may impact in their performance. In this work, the long-term effects of TiO2 NPs on the main benefits of treating wastewater by UASB reactors such as, biogas production, methane fraction in biogas and organic matter removal were evaluated. Evaluation consisted of monitoring such parameters in two identical UASB reactors, one UASB-Control (without NPs) and the experimental one (UASB-TiO2 NPs) that received wastewater with TiO2 NPs. The fate of NPs in the UASB reactor was also determined. Results indicated that biogas production increased by 8.8% due to the chronic exposure of UASB reactor to TiO2 NPs during the first 44 days of experiment. However, the methane content in the biogas had no significant differences between both UASB, ranging between 78% and 90% of methane during the experiment. The removal of organic matter in both UASB was similar and ranged 92-98% along the experimental time. This means that accumulation of TiO2 NPs did not altered the biogas production and organic matter removal. However, the content of total volatile solids (TVS) in UASB-TiO2 NPs dropped off from 137.8 g to 64.2 g in 84 days, while for control reactor that decreased from 141.6 g to 92.4 g in the same period. Hence, the increased biogas production in the UASB exposed to TiO2 was attributed to hydrolysis of the TVS in this reactor. The main fate of TiO2 NPs was the granular sludge, which accumulated up to 8.56 mg Ti/g, which represent around 99% of the Ti spiked to the reactor and the possible cause of the biomass hydrolyzation in the UASB. Disposal of UASB sludge containing NPs from may raise ecotoxicological concerns due to the use of biosolids in agricultural activities.

Nitrification of anaerobic digestate using a consortium of microalgae and nitrifiers in an open photobioreactor with moving bed carriers.

A consortium of microalgae and nitrifiers has attracted attention as an alternative to the expensive traditional nitrification process. A possible obstacle to achieving this is the inhibition of nitrifiers under strong light irradiation. This study evaluated the effect of moving bed carriers on anaerobic digestate nitrification in an open photobioreactor inoculated with microalgae and nitrifiers under an incident light intensity of 1000 µmol photons m-2 s-1. The results showed higher specific nitrification activity in the carrier-added photobioreactor (103.6 mg-N g-TSS day-1) than in one in which no carrier was added (11.7 mg-N g-TSS day-1). The empirical equations for determining the light intensity at different depths in the photobioreactor showed a significant contribution by carriers in attenuating the incident light intensity. This is due to the large light attenuation caused by the carrier (1.09 cm-1). The average light intensity inside of the photobioreactor decreased considerably in the carrier-added photobioreactor (342 µmol photons m-2 s-1), whereas it did not decrease in the one with no added carrier. It was found that specific nitrification activity was significantly negatively affected by average light intensity inside of the reactor, and not by incident light intensity, by combining the results from different studies including ours. This study demonstrated, for the first time, the effectiveness of adding moving bed carriers in photobioreactors to mitigate light inhibition of nitrifiers in a consortium of microalgae and nitrifiers.

Anaerobic digestion effluent treatment using microalgae and nitrifiers in an outdoor raceway pond with fluidized carriers.

Combining microalgae and nitrifiers in a single photobioreactor has attracted attention as an alternative approach for conventional nitrogen removal from wastewater. However, nitrifiers are known to be sensitive to light exposure. This study demonstrated the effectiveness of using fluidized carriers to mitigate light stress in nitrifiers. An outdoor raceway pond containing microalgae and nitrifiers with fluidized carriers was used to treat two-fold diluted anaerobic digestion effluent (785 mg-N L-1 as a form of dissolved total Kjeldahl nitrogen: TKN) over 50 days. The average daily sunlight intensity reached the inhibition level of nitrifiers (423 µmol photons m-2 s-1); however, stable nitrification with a specific ammonium oxidation rate of 55 mg-N g-total suspended solid-1 day-1 was observed. TKN was mostly removed via nitrifier metabolism (ammonium oxidation and uptake: 40.1%) and partially via microalgae uptake (5.7%). Different microalgae-based processes including that of this study were compared in terms of tolerances to a high dissolved TKN concentration and strong light. Our system showed a relatively higher resistance to not only light exposure but also TKN because the nitrification process decreased the free ammonia level to less than 0.25 mg L-1, which allowed microalgae to grow despite the high ammonium concentration.

Effects of different light conditions on ammonium removal in a consortium of microalgae and partial nitrifying granules.

Ammonium removal by a coupling process of microalgae (Chlorella sorokiniana) with partial nitrifying granules was evaluated in batch reactors illuminated in a wide range of light intensities (0, 100, 450, and 1600 µmol photons m-2 s-1). Ammonium oxidation performance for different light exposure time showed that the granules had a light stress tolerance at 1600 µmol photons m-2 s-1 for up to 12 h, but continuous illumination induced severe inhibition on nitrifying bacteria thereafter. Ammonium removal efficiencies at the end of tests were 66%, 62%, 5%, and -10% (due to ammonification) for 0, 100, 450, and 1600 µmol photons m-2 s-1, respectively. The nitrogen mass balance shows co-occurrence of microalgal growth taking up 24% of fed ammonium and nitrifying bacteria oxidizing 38% of fed ammonium at 100 µmol photons m-2 s-1, while both nitrification and microalgal growth are inhibited at light intensity above 450 µmol photons m-2 s-1. In comparing results from this study with previous results, it was found that the ammonium removal pathway, i.e., nitrification or microalgal uptake, is regulated more strongly by daily average light intensity than by instantaneous light intensity. Empirical model equations to estimate the oxygen balance in consortium reactors categorized the effect of daily average light intensities on process performance as follows: (i) below 27 µmol photons m-2 s-1: insufficient oxygen for nitrification; (ii) 27 to 35: sufficient oxygen for nitrification via nitrite; (iii) 35 to 180: sufficient oxygen for nitrification via nitrate; (iv) above approximately 200-300: oversaturated dissolved oxygen, excess free ammonia and/or intensive light inhibitions.

Effects and fate of TiO2 nanoparticles in the anaerobic treatment of wastewater and waste sludge.

The increasing use of TiO2 nanoparticles (NPs) in customer products has also increased the concerns about their effects in the environment. Anaerobic digestion is a process probably exposed to high concentrations of TiO2 NPs due to its application for wastewater and waste sludge treatment. In this work, it was studied the anaerobic digestion performance and the extracellular polymeric substances (EPS) production in presence of TiO2 NPs, as well as the fate of TiO2 NPs in anaerobic reactors. Results showed that methane production enhanced an average of 14.9% in presence TiO2 NPs, which is considered a positive effect. A strong affinity between TiO2 NPs and EPS was found, especially for proteins (PRO) and polysaccharides (PS) in the loosely and tightly bound EPS layers of microorganisms (LB-EPS and TB-EPS). Ti quantification indicated that 92% of the TiO2 NPs are removed by anaerobic sludge, while 8% remain in the treated effluent.

Desulfonatronum parangueonense sp. nov., a sulfate-reducing bacterium isolated from sediment of an alkaline crater lake.

Novel Gram-stain-negative, non-spore-forming, vibrio-shaped, anaerobic, alkaliphilic, sulfate-reducing bacteria, designated strains PAR180T and PAR190, were isolated from sediments collected at an alkaline crater lake in Guanajuato (Mexico). Strain PAR180T grew at temperatures between 15 and 40 °C (optimum 35 °C), and at pH between 8.3 and 10.4 (optimum 9). It was halotolerant, growing with up to 8 % (w/v) NaCl. Lactate, formate, pyruvate and ethanol were used as electron donors in the presence of sulfate and were incompletely oxidized to acetate and CO2. The isolate was able to grow with hydrogen and with CO2 as a carbon source. Beside sulfate, sulfite and thiosulfate were used as terminal electron acceptors. The isolate was able to grow by disproportionation of sulfite and thiosulfate, but not elemental sulfur, using acetate as a carbon source. The predominant fatty acids were C16 : 0, C16 : 1ω7c and summed feature 10 (C18 : 1ω7c and/or C18 : 1ω9t and/or C18 : 1ω12t). The DNA G+C content was 56.1 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that it belongs to the genus Desulfonatronum, class Deltaproteobacteria. Its closest relative is Desulfonatronum thiosulfatophilum (98.7 % 16S rRNA gene sequence similarity). The DNA-DNA relatedness value between strain PAR180T and the type strain of D. thiosulfatophilum was 37.1±2.5 %. On the basis of phylogenetic, phenotypic and chemotaxonomic characteristics, the isolates is considered to represent a novel species of the genus Desulfonatronum, for which the name Desulfonatronum parangueonense sp. nov. is proposed. The type strain is PAR180T (=DSM 103602T=JCM 31598T).

Interactions and effects of metal oxide nanoparticles on microorganisms involved in biological wastewater treatment.

To clarify the toxicological effects of metal oxide nanoparticles (NPs) on microorganisms with environmental relevance, it is necessary to understand their interactions. In this work, they were studied the effects and the morphological interactions of two metal oxide NPs (ZnO and TiO2 ) with microorganisms, during aerobic treatment of wastewater. The effects were evaluated according to nutrient removal from wastewater, while morphological interactions were determined by three different techniques such as TEM, HAADF-STEM, as well as an elemental mapping. According to results about effects of both NPs, they inhibited the removal of organic matter and ammonia nitrogen, and enhanced the orthophosphate removal. Related to morphological interactions, the electron-dense material of both NPs was mainly observed bounded to cell membrane. In tests with ZnO NPs, it was also observed electron-dense material internalized in microorganisms without physical damage in cell membrane. The elemental mapping was useful to determine that the electron-dense material corresponded to Zn and Ti. Both interactions, internalization and attachment of NPs on cell membrane of microorganisms may trigger the negative effect in the removal of organic matter and nitrogen.

Changes in nutrient removal and flocs characteristics generated by presence of ZnO nanoparticles in activated sludge process.

The aim of this work was to evaluate the impact generated by ZnO NPs on the activated sludge process treating raw (RWW) and filtered wastewater (FWW). It was analyzed the oxygen uptake rate, nutrient removal, flocs characteristics and the morphological interactions between activated sludge and ZnO NPs, in presence of 450-2000 mg/L. The results showed that the presence of more than 450 mg/L of ZnO NPs in raw and filtered wastewater inhibited the oxygen uptake by activated sludge. The highest inhibition was 35% in presence of 1500 mg/L in RWW. The organic matter removal was only inhibited in the presence of 450 and 900 mg/L of ZnO NPs; while ammonia removal decreased for all concentrations of ZnO NPs in both types of wastewater, around 13% for RWW and up to 9% for FWW. The orthophosphate removal improved as the concentration of ZnO NPs increased for both wastewater types, enhancing up to 8% for RWW and 17% for FWW. The flocs size of activated sludge exposed to ZnO NPs in RWW decreased as the concentration of ZnO NPs increased; while for FWW, an opposite effect was observed. The elemental mapping allowed detect the Zn inside of microorganisms, which may correspond to a toxicity mechanism in RWW and FWW. These results indicated that the changes in nutrient removal and flocs characteristics caused by the presence of ZnO NPs on the activated sludge are related to wastewater characteristics, such as suspended solids, type of substrate and concentration of ZnO NPs.

Influence of wastewater type on the impact generated by TiO2 nanoparticles on the oxygen uptake rate in activated sludge process.

Physicochemical characteristics of wastewater have a relationship with the stability of TiO2 nanoparticles (NPs). This in turn has an effect on the toxicity of TiO2 NPs in microorganisms. In this work, the effect of TiO2 NPs on activated sludge process was evaluated using three different types of wastewater: synthetic, raw, and filtered. The results showed that aggregate size of TiO2 NPs and their specific adsorption of substrates were influenced by the type of substrates and the presence of suspended solids in the wastewater. It was also shown that TiO2 NPs in raw wastewater severely inhibited oxygen uptake by microorganisms as compared to uptake in synthetic or filtered wastewater. The attachment of TiO2 NP aggregates on cell membranes was observed for all types of wastewater. However, the internalization of TiO2 NPs by microorganisms was observed only for raw and filtered wastewater. These results indicate that the effects caused by TiO2 NPs on activated sludge were different depending on the wastewater used for the experiment.

Impact of hydrocarbons, PCBs and heavy metals on bacterial communities in Lerma River, Salamanca, Mexico: Investigation of hydrocarbon degradation potential.

Freshwater contamination usually comes from runoff water or direct wastewater discharges to the environment. This paper presents a case study which reveals the impact of these types of contamination on the sediment bacterial population. A small stretch of Lerma River Basin, heavily impacted by industrial activities and urban wastewater release, was studied. Due to industrial inputs, the sediments are characterized by strong hydrocarbon concentrations, ranging from 2 935 to 28 430µg·kg(-1) of total polyaromatic hydrocarbons (PAHs). These sediments are also impacted by heavy metals (e.g., 9.6µg·kg(-1) of Cd and 246µg·kg(-1) of Cu, about 8 times the maximum recommended values for environmental samples) and polychlorinated biphenyls (ranging from 54 to 123µg·kg(-1) of total PCBs). The bacterial diversity on 6 sediment samples, taken from upstream to downstream of the main industrial and urban contamination sources, was assessed through TRFLP. Even though the high PAH concentrations are hazardous to aquatic life, they are not the only factor driving bacterial community composition in this ecosystem. Urban discharges, leading to hypoxia and low pH, also strongly influenced bacterial community structure. The bacterial bioprospection of these samples, using PAH as unique carbon source, yielded 8 hydrocarbonoclastic strains. By sequencing the 16S rDNA gene, these were identified as similar to Mycobacterium goodii, Pseudomonas aeruginosa, Pseudomonas lundensis or Aeromonas veronii. These strains showed high capacity to degrade naphthalene (between 92 and 100% at 200mg·L(-1)), pyrene (up to 72% at 100mg·L(-1)) and/or fluoranthene (52% at 50mg·L(-1)) as their only carbon source on in vitro experiments. These hydrocarbonoclastic bacteria were detected even in the samples upstream of the city of Salamanca, suggesting chronical contamination, already in place longer before. Such microorganisms are clearly potential candidates for hydrocarbon degradation in the treatment of oil discharges.

Microbial diversity in Los Azufres geothermal field (Michoacán, Mexico) and isolation of representative sulfate and sulfur reducers.

Los Azufres spa consists of a hydrothermal spring system in the Mexican Volcanic Axis. Five samples (two microbial mats, two mud pools and one cenote water), characterized by high acidity (pH between 1 and 3) and temperatures varying from 27 to 87 °C, were investigated for their microbial diversity by Terminal-Restriction Fragment Length Polymorphism (T-RFLP) and 16S rRNA gene library analyses. These data are the first to describe microbial diversity from Los Azufres geothermal belt. The data obtained from both approaches suggested a low bacterial diversity in all five samples. Despite their proximity, the sampling points differed by their physico-chemical conditions (mainly temperature and matrix type) and thus exhibited different dominant bacterial populations: anoxygenic phototrophs related to the genus Rhodobacter in the biomats, colorless sulfur oxidizers Acidithiobacillus sp. in the warm mud and water samples, and Lyzobacter sp.-related populations in the hot mud sample (87 °C). Molecular data also allowed the detection of sulfate and sulfur reducers related to Thermodesulfobium and Desulfurella genera. Several strains affiliated to both genera were enriched or isolated from the mesophilic mud sample. A feature common to all samples was the dominance of bacteria involved in sulfur and iron biogeochemical cycles (Rhodobacter, Acidithiobacillus, Thiomonas, Desulfurella and Thermodesulfobium genera).

Treatment of a textile effluent from dyeing with cochineal extracts using Trametes versicolor fungus.

Trametes versicolor (Tv) fungus can degrade synthetic dyes that contain azo groups, anthraquinone, triphenylmethane polymers, and heterocyclic groups. However, no references have been found related to the degradation of natural dyes, such as the carminic acid that is contained in the cochineal extract. Experiments to determine the decolorization of the effluent used in the cotton dyeing process with cochineal extract by means of Tv fungus were done. Treatments to determine decolorization in the presence or absence of Kirk's medium, glucose, and fungus, with an addition of 50% (v v-1) of nonsterilized effluent were performed. Physicochemical characterization was performed at the start and end of the treatment. Degradation kinetics were determined. A direct relationship was found between the dry weight of fungi, pH, and the decolorization system, with higher decolorization at lower pH levels (pH ~4.3). High decolorization (81% ± 0.09; 88% ± 0.17; and 99% ± 0.04) for three of the eight treatments (Kirk's medium without glucose, Kirk's medium with glucose, and without medium with glucose, respectively) was found. Toxicity tests determined an increase in the initial effluent toxicity (7.33 TU) compared with the final treatment (47.73 TU) in a period of 11 days. For this system, a degradation sequence of the carminic acid structure present in the effluent by the Tv fungus is suggested, in which it is seen that metabolites still containing aromatic structures are generated.