Design and Validation of Primer Sets for the Detection and Quantification of Antibiotic Resistance Genes in Environmental Samples by Quantitative PCR.

The high prevalence of antibiotic resistant bacteria (ARB) in several environments is a great concern threatening human health. Particularly, wastewater treatment plants (WWTP) become important contributors to the dissemination of ARB to receiving water bodies, due to the inefficient management or treatment of highly antibiotic-concentrated wastewaters. Hence, it is vital to develop molecular tools that allow proper monitoring of the genes encoding resistances to these important therapeutic compounds (antibiotic resistant genes, ARGs). For an accurate quantification of ARGs, there is a need for sensitive and robust qPCR assays supported by a good design of primers and validated protocols. In this study, eleven relevant ARGs were selected as targets, including aadA and aadB (conferring resistance to aminoglycosides); ampC, blaTEM, blaSHV, and mecA (resistance to beta-lactams); dfrA1 (resistance to trimethoprim); ermB (resistance to macrolides); fosA (resistance to fosfomycin); qnrS (resistance to quinolones); and tetA(A) (resistance to tetracyclines). The in silico design of the new primer sets was performed based on the alignment of all the sequences of the target ARGs (orthology grade > 70%) deposited in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, allowing higher coverages of the ARGs' biodiversity than those of several primers described to date. The adequate design and performance of the new molecular tools were validated in six samples, retrieved from both natural and engineered environments related to wastewater treatment. The hallmarks of the optimized qPCR assays were high amplification efficiency (> 90%), good linearity of the standard curve (R2 > 0.980), repeatability and reproducibility across experiments, and a wide linear dynamic range. The new primer sets and methodology described here are valuable tools to upgrade the monitorization of the abundance and emergence of the targeted ARGs by qPCR in WWTPs and related environments.

Microbial Ecology of Granular Biofilm Technologies for Wastewater Treatment: A Review.

Nowadays, the discharge of wastewater is a global concern due to the damage caused to human and environmental health. Wastewater treatment has progressed to provide environmentally and economically sustainable technologies. The biological treatment of wastewater is one of the fundamental bases of this field, and the employment of new technologies based on granular biofilm systems is demonstrating success in tackling the environmental issues derived from the discharge of wastewater. The granular-conforming microorganisms must be evaluated as functional entities because their activities and functions for removing pollutants are interconnected with the surrounding microbiota. The deep knowledge of microbial communities allows for the improvement in system operation, as the proliferation of microorganisms in charge of metabolic roles could be modified by adjustments to operational conditions. This is why engineering must consider the intrinsic microbiological aspects of biological wastewater treatment systems to obtain the most effective performance. This review provides an extensive view of the microbial ecology of biological wastewater treatment technologies based on granular biofilms for mitigating water pollution.

Anticancer drugs impact the performance and prokaryotic microbiome of an aerobic granular sludge system operated in a sequential batch reactor.

Increased concerns exist about the presence of anticancer drugs in wastewater. However, knowledge of the impacts of anticancer drugs on the performance of the system and microbial communities during wastewater treatment processes is limited. We examined the effect of three anticancer drugs commonly detected in influents of wastewater treatment plants applied at three different concentration levels on the performance, efficiency of anticancer drug removal, and prokaryotic microbiome in an aerobic granular sludge system (AGS) operated in a sequential batch reactor (SBR). We showed that an AGS can efficiently remove anticancer drugs, with removal rates in the range of 53-100% depending on the type of drug and concentration level. Anticancer drugs significantly decreased the abundance of total bacterial and archaeal communities, an effect that was linked to reduced nitrogen removal efficiency. Anticancer drugs also reduced the diversity, altered the prokaryotic community composition, reduced network complexity, and induced a decrease of a wide range of predicted bacterial functions. Specific bacterial taxa responsive to the addition of anticancer drugs with known roles in nitrification and denitrification were identified. This study shows anticancer drugs should be monitored in the future as they can induce changes in the performance and microbiome of wastewater treatment technologies.

Promising bioprocesses for the efficient removal of antibiotics and antibiotic-resistance genes from urban and hospital wastewaters: Potentialities of aerobic granular systems.

The use, overuse, and improper use of antibiotics have resulted in higher levels of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs), which have profoundly disturbed the equilibrium of the environment. Furthermore, once antibiotic agents are excreted in urine and feces, these substances often can reach wastewater treatment plants (WWTPs), in which improper treatments have been highlighted as the main reason for stronger dissemination of antibiotics, ARB, and ARGs to the receiving bodies. Hence, achieving better antibiotic removal capacities in WWTPs is proposed as an adequate approach to limit the spread of antibiotics, ARB, and ARGs into the environment. In this review, we highlight hospital wastewater (WW) as a critical hotspot for the dissemination of antibiotic resistance due to its high level of antibiotics and pathogens. Hence, monitoring the composition and structure of the bacterial communities related to hospital WW is a key factor in controlling the spread of ARGs. In addition, we discuss the advantages and drawbacks of the current biological WW treatments regarding the antibiotic-resistance phenomenon. Widely used conventional activated sludge technology has proved to be ineffective in mitigating the dissemination of ARB and ARGs to the environment. However, aerobic granular sludge (AGS) technology is a promising technology-with broad adaptability and excellent performance-that could successfully reduce antibiotics, ARB, and ARGs in the generated effluents. We also outline the main operational parameters involved in mitigating antibiotics, ARB, and ARGs in WWTPs. In this regard, WW operation under long hydraulic and solid retention times allows better removal of antibiotics, ARB, and ARGs independently of the WW technology employed. Finally, we address the current knowledge of the adsorption and degradation of antibiotics and their importance in removing ARB and ARGs. Notably, AGS can enhance the removal of antibiotics, ARB, and ARGs due to the complex microbial metabolism within the granular biomass.

Novel insights into the impact of anticancer drugs on the performance and microbial communities of a continuous-flow aerobic granular sludge system.

Anticancer drugs are frequently found in domestic wastewater, but knowledge of their impacts on wastewater treatment processes is limited. The effects of three levels of concentrations (low, medium, and high) of three anticancer drugs on physicochemical parameters and prokaryotic communities of a continuous-flow aerobic granular sludge (AGS) system were examined. Drugs at medium and high concentrations reduced the removal of total nitrogen and organic matter during the first 15 days of operation by approximately 15-20 % compared to a control, but these effects disappeared afterward. Removal efficiencies of drugs were in the range of 51.2-100 % depending on the concentration level. Drugs at medium and high concentrations reduced the abundance and diversity and altered the composition of prokaryotic communities. Specific taxa were linked to variations in performance parameters after the addition of the drugs. This study provides improved knowledge of the impacts of anticancer drugs in AGS systems operated in continuous-flow reactor.

Deciphering the Role of WWTPs in Cold Environments as Hotspots for the Dissemination of Antibiotic Resistance Genes.

Cold environments are the most widespread extreme habitats in the world. However, the role of wastewater treatment plants (WWTPs) in the cryosphere as hotspots in antibiotic resistance dissemination has not been well established. Hence, a snapshot of the resistomes of WWTPs in cold environments, below 5 °C, was provided to elucidate their role in disseminating antibiotic resistance genes (ARGs) to the receiving waterbodies. The resistomes of two natural environments from the cold biosphere were also determined. Quantitative PCR analysis of the aadA, aadB, ampC, blaSHV, blaTEM, dfrA1, ermB, fosA, mecA, qnrS, and tetA(A) genes indicated strong prevalences of these genetic determinants in the selected environments, except for the mecA gene, which was not found in any of the samples. Notably, high abundances of the aadA, ermB, and tetA(A) genes were found in the influents and activated sludge, highlighting that WWTPs of the cryosphere are critical hotspots for disseminating ARGs, potentially worsening the resistance of bacteria to some of the most commonly prescribed antibiotics. Besides, the samples from non-disturbed cold environments had large quantities of ARGs, although their ARG profiles were highly dissimilar. Hence, the high prevalences of ARGs lend support to the fact that antibiotic resistance is a common issue worldwide, including environmentally fragile cold ecosystems.

Anticancer drugs drive changes in the performance, abundance, diversity, and composition of eukaryotic communities of an aerobic granular sludge system.

Anticancer drugs are emerging contaminants that are being increasingly detected in urban wastewater. However, there is limited knowledge on the use of biological wastewater treatments, such as granular sludge systems (AGSs), to remove these substances and on their impacts on the general performance of the system and the eukaryotic communities in the granules. We investigated the impacts of three anticancer drugs commonly found in wastewater treatment plants and applied at three different concentrations on the removal efficiency of anticancer drugs, physicochemical parameters, and the eukaryotic microbiome of an AGS operated in a sequential batch reactor (SBR). Anticancer drugs applied at medium and high concentrations significantly decreased the removal efficiency of total nitrogen, the granular biomass concentration, and the size and setting velocity of granules. However, these effects disappeared after not adding the drugs for about a month thus showing the plasticity of the system to return to original levels. Regardless of the concentration of anticancer drugs tested, the AGS technology was effective in removing these substances, with removal rates in the range of 68.5%-100%. The presence of anticancer drugs at medium and high concentrations significantly decreased the abundance of total fungi, an effect that was linked to changes in the physicochemical parameters. Anticancer drugs also induced decreases in the diversity of the eukaryotic community, altered the community composition, and reduced the network complexity when applied at medium and high concentrations. Taxa responsive to the presence of anticancer drugs were identified. The diversity and composition of the eukaryotic microbiome returned to original diversity levels after not adding the drugs for about a month. Overall, this study increases our understanding of the impacts of anticancer drugs on the performance and eukaryotic microbiome of an AGS and highlights the need for monitoring these substances.

Anticancer drugs in wastewater and natural environments: A review on their occurrence, environmental persistence, treatment, and ecological risks.

The consumption of anticancer drugs (also known as chemotherapy drugs or antineoplastic drugs) has augmented over the last decades due to increased cancer incidence. Although there is an increasing concern about the presence of pharmaceutical compounds in natural environments and urban/domestic wastewater, anticancer drugs used in chemotherapy and anticancer medication have received less attention. In this review, the occurrence, environmental persistence, and known and potential ecological impacts of anticancer drugs is discussed. This review shows that these compounds are being increasingly detected in effluents of hospitals, influents and effluents of wastewater treatment plants, river surface water and sediments, groundwater, and even drinking water. Anticancer drugs can impact aquatic organisms such as algae, crustaceans, rotifers, and fish and may promote changes in soil and water microbial communities that may alter ecosystem functioning. Our knowledge of technologies for the removal of anticancer drugs is still limited, and these drugs can be dispersed in nature in a diffuse way in an uncontrolled manner. For this reason, an improved understanding of the presence, persistence, and ecological impacts of anticancer drugs in wastewater and natural environments is needed to help design management strategies, protect aquatic microorganisms, and mitigate potential ecological impacts.

Structure of fungal communities in sequencing batch reactors operated at different salinities for the selection of triacylglyceride-producers from a fish-canning lipid-rich waste stream.

Oleaginous fungi natively accumulate large amounts of triacylglycerides (TAG), widely used as precursors for sustainable biodiesel production. However, little attention has been paid to the diversity and roles of fungal mixed microbial cultures (MMCs) in sequencing batch reactors (SBR). In this study, a lipid-rich stream produced in the fish-canning industry was used as a substrate in two laboratory-scale SBRs operated under the feast/famine (F/F) regime to enrich microorganisms with high TAG-storage ability, under two different concentrations of NaCl (SBR-N: 0.5 g/L; SBR-S: 10 g/L). The size of the fungal community in the enriched activated sludge (EAS) was analyzed using 18S rRNA-based qPCR, and the fungal community structure was determined by Illumina sequencing. The different selective pressures (feeding strategy and control of pH) implemented in the enrichment SBRs throughout operation increased the abundance of total fungi. In general, there was an enrichment of genera previously identified as TAG-accumulating fungi (Apiotrichum, Candida, Cutaneotrichosporon, Geotrichum, Haglerozyma, Metarhizium, Mortierella, Saccharomycopsis, and Yarrowia) in both SBRs. However, the observed increase of their relative abundances throughout operation was not significantly linked to a higher TAG accumulation.

Spontaneous pneumopericardium and pneumomediastinum as atypical complication for SARS-CoV-2.

SARS-CoV-2 (COVID-19) disease is an infection caused by a new emerging coronavirus, the most common clinical manifestations include fever, dry cough, dyspnea, chest pain, fatigue, and myalgia, sometimes it may present with atypical manifestations such as spontaneous pneumothorax and pneumomediastinum that occur in a minority of patients. We report a case of spontaneous pneumopericardium in a 60-year-old male, without comorbidities or a history of trauma, with pneumonia due to SARS-CoV-2.

La enfermedad por SARS-CoV-2 (COVID-19) es una infección causada por un nuevo coronavirus emergente. Las manifestaciones clínicas más comunes incluyen fiebre, tos seca, disnea, dolor de pecho, fatiga y mialgias. En ocasiones puede presentarse con manifestaciones atípicas, como neumotórax espontáneo y neumomediastino, que ocurren en una minoría de pacientes. Reportamos un caso de neumopericardio espontáneo en un varón de 60 años, sin comorbilidad ni antecedente de traumatismo, con neumonía por SARS-CoV-2.

Pharmaceutical Pollution in Aquatic Environments: A Concise Review of Environmental Impacts and Bioremediation Systems.

The presence of emerging contaminants in the environment, such as pharmaceuticals, is a growing global concern. The excessive use of medication globally, together with the recalcitrance of pharmaceuticals in traditional wastewater treatment systems, has caused these compounds to present a severe environmental problem. In recent years, the increase in their availability, access and use of drugs has caused concentrations in water bodies to rise substantially. Considered as emerging contaminants, pharmaceuticals represent a challenge in the field of environmental remediation; therefore, alternative add-on systems for traditional wastewater treatment plants are continuously being developed to mitigate their impact and reduce their effects on the environment and human health. In this review, we describe the current status and impact of pharmaceutical compounds as emerging contaminants, focusing on their presence in water bodies, and analyzing the development of bioremediation systems, especially mycoremediation, for the removal of these pharmaceutical compounds with a special focus on fungal technologies.

Effects of sulphur amino acids on the size and structure of microbial communities of aerobic granular sludge bioreactors.

Granular activated sludge has been described as a promising tool in treating wastewater. However, the effect of high concentrations of sulphur amino acids, cysteine and methionine, in the evolution, development and stability of AGS-SBRs (aerobic granular sludge in sequential batch reactors) and their microbial communities is not well-established. Therefore, this study aimed to evaluate microbial communities' size, structure and dynamics in two AGS-SBRs fed with two different concentrations of amino acids (50 and 100 mg L-1 of both amino acids). In addition, the impact of the higher level of amino acids was also determined under an acclimatization or shock strategy. While N removal efficiency decreased with amino acids, the removal of the organic matter was generally satisfactory. Moreover, the abrupt presence of both amino acids reduced even further the removal performance of N, whereas under progressive adaptation, the removal yield was higher. Besides, excellent removal rates of cysteine and methionine elimination were found, in all stages below 80% of the influent values. Generally considered, the addition of amino acids weakly impacts the microbial communities' total abundances. On the contrary, the presence of amino acids sharply modulated the dominant bacterial structures. Furthermore, the highest amino acid concentration under the shock strategy resulted in a severe change in the structure of the microbial community. Acidovorax, Flavobacterium, Methylophilus, Stenotrophomonas and Thauera stood out as the prominent bacteria to cope with the high presence of cysteine and methionine. Hence, the AGS-SBR technology is valuable for treating influents enriched in sulphur Aa inclusively when a shock strategy was used.

Dynamics of PHA-Accumulating Bacterial Communities Fed with Lipid-Rich Liquid Effluents from Fish-Canning Industries.

The biosynthesis of polyhydroxyalkanoates (PHAs) from industrial wastes by mixed microbial cultures (MMCs) enriched in PHA-accumulating bacteria is a promising technology to replace petroleum-based plastics. However, the populations' dynamics in the PHA-accumulating MMCs are not well known. Therefore, the main objective of this study was to address the shifts in the size and structure of the bacterial communities in two lab-scale sequencing batch reactors (SBRs) fed with fish-canning effluents and operated under non-saline (SBR-N, 0.5 g NaCl/L) or saline (SBR-S, 10 g NaCl/L) conditions, by using a combination of quantitative PCR and Illumina sequencing of bacterial 16S rRNA genes. A double growth limitation (DGL) strategy, in which nitrogen availability was limited and uncoupled to carbon addition, strongly modulated the relative abundances of the PHA-accumulating bacteria, leading to an increase in the accumulation of PHAs, independently of the saline conditions (average 9.04 wt% and 11.69 wt%, maximum yields 22.03 wt% and 26.33% SBR-N and SBR-S, respectively). On the other hand, no correlations were found among the PHAs accumulation yields and the absolute abundances of total Bacteria, which decreased through time in the SBR-N and did not present statistical differences in the SBR-S. Acinetobacter, Calothrix, Dyella, Flavobacterium, Novosphingobium, Qipengyuania, and Tsukamurella were key PHA-accumulating genera in both SBRs under the DGL strategy, which was revealed as a successful tool to obtain a PHA-enriched MMC using fish-canning effluents.

Agrobacterium leguminum sp. nov., isolated from nodules of Phaseolus vulgaris in Spain.

Two endophytic strains, coded MOVP5T and MOPV6, were isolated from nodules of Phaseolus vulgaris plants grown on agricultural soil in Southeastern Spain, and were characterized through a polyphasic taxonomy approach. Their 16S rRNA gene sequences showed 99.3 and 99.4 %, 98.9 and 99.6 %, and 99.0 and 98.7% similarity to 'A. deltaense' YIC 4121T, A. radiobacter LGM 140T, and A. pusense NRCPB10T, respectively. Multilocus sequence analysis based on sequences of recA and atpD genes suggested that these two strains could represent a new Agrobacterium species with less than 96.5 % similarity to their closest relatives. PCR amplification of the telA gene, involved in synthesis of protelomerase, confirmed the affiliation of strains MOPV5T and MOPV6 to the genus Agrobacterium. Whole genome average nucleotide identity and digital DNA-DNA hybridization average values were less than 95.1 and 66.7 %, respectively, with respect to its closest related species. Major fatty acids in strain MOPV5T were C18 : 1 ω7c/C18 : 1 ω6c in summed feature 8, C19 : 0 cyclo ω8c, C16 : 0 and C16 : 0 3-OH. Colonies were small to medium, pearl-white coloured on YMA at 28 °C and growth was observed at 10-42 °C, pH 5.0-10.0 and with 0.0-0.5 % (w/v) NaCl. The DNA G+C content was 59.9 mol%. These two strains differ from all other genomovars of Agrobacterium found so far, including those that have not yet given a Latin name. The combined genotypic, phenotypic and chemotaxonomic data support the classification of strain MOPV5T as representing a novel species of Agrobacterium, for which the name Agrobacterium leguminum sp. nov. is proposed. The type strain is MOPV5T (=CECT 30096T=LMG 31779T).

Design of Bio-Absorbent Systems for the Removal of Hydrocarbons from Industrial Wastewater: Pilot-Plant Scale.

The objective of this study was the development and design of a treatment system at a pilot-plant scale for the remediation of hydrocarbons in industrial wastewater. The treatment consists of a combined approach of absorption and biodegradation to obtain treated water with sufficient quality to be reused in fire defense systems (FDSs). The plant consists of four vertical flow columns (bioreactors) made of stainless steel (ATEX Standard) with dimensions of 1.65 × 0.5 m and water volumes of 192.4 L. Each bioreactor includes a holder to contain the absorbent material (Pad Sentec polypropylene). The effectiveness of the treatment system has been studied in wastewater with high and low pollutant loads (concentrations higher than 60,000 mg L-1 of total petroleum hydrocarbons (TPH) and lower than 500 mg L-1 of TPHs, respectively). The pilot-plant design can function at two different flow rates, Q1 (180 L h-1) and Q2 (780 L h-1), with or without additional aeration. The results obtained for strongly polluted wastewaters showed that, at low flow rates, additional aeration enhanced hydrocarbon removal, while aeration was unnecessary at high flow rates. For wastewater with a low pollutant load, we selected a flow rate of 780 L h-1 without aeration. Different recirculation times were also tested along with the application of a post-treatment lasting 7 days inside the bioreactor without recirculation. The microbial diversity studies showed similar populations of bacteria and fungi in the inlet and outlet wastewater. Likewise, high similarity indices were observed between the adhered and suspended biomass within the bioreactors. The results showed that the setup and optimization of the reactor represent a step forward in the application of bioremediation processes at an industrial/large scale.

High triglyceride to HDL-cholesterol ratio as a biochemical marker of severe outcomes in COVID-19 patients.

BACKGROUND & AIMS: Coronavirus disease 2019 (COVID-19) patients with severe complications have shown comorbidities with cardiovascular-disease, hypertension and type 2 diabetes mellitus; clinical disorders that share the common metabolic alterations of insulin resistance and dyslipidaemia. A high triglyceride to high density lipoprotein cholesterol (Tg/HDL c) ratio has been associated with reduced insulin sensitivity, metabolic syndrome and adverse cardiovascular events. Our aim in this study was to determine the association between different components of the lipid profile and particularly the Tg/HDL c ratio with severe complications like the requirement of invasive mechanical ventilation in COVID-19 patients. METHODS: We collected demographic, clinical and biochemical data to conduct a cohort study in 43 adult patients with confirmed COVID-19 diagnosis by quantitative polymerase chain reaction (qPCR) at baseline and in the subsequent 15 days. Patients were subjected to a very similar treatment scheme with the JAK1/2 inhibitor ruxolitinib. Descriptive statistics, variable association and logistic regression were applied to identify predictors of disease severity among elements and calculations from the lipid profile. RESULTS: Patients were aged 57 ± 14 years; 55.8% were male from which 75% required hospitalization and 44.2% were female who 58% were hospitalized. The most common comorbidities were type 2 diabetes mellitus (58%) and hypertension (40%). Hospitalized and critical care patients showed lower HDL c blood levels and increased Tg/HDL c ratio than those with outpatient management and mild/asymptomatic COVID-19. Tg/HDL c ratio correlated with variables of disease severity such as lactate dehydrogenase (LDH) levels (r = 0.356; p < 0.05); National Early Warning Score 2 (NEWS 2) (r = 0.495; p < 0.01); quick sequential organ failure assessment (qSOFA) (r = 0.538; p < 0.001); increased need of oxygen support (r = 0.447; p < 0.01) and requirement of mechanical ventilation (r = 0.378; p < 0.05). Tg/HDL c ratio had a negative correlation with partial oxygen saturation/fraction of inspired oxygen (SaO 2/FiO2) ratio (r = -0.332;p < 0.05). Linear regression analysis showed that Tg/HDL c ratio can predict increases in inflammatory factors like LDH (p < 0.01); ferritin (p < 0.01) and D-dimer (p < 0.001). Logistic regression model indicated that ≥7.45 Tg/HDL c ratio predicts requirement of invasive mechanical ventilation (OR 11.815, CI 1.832-76.186, p < 0.01). CONCLUSIONS: The Tg/HDLc ratio can be used as an early biochemical marker of COVID-19 severe prognosis with requirement of invasive mechanical ventilation.

Salinity is the major driver of the global eukaryotic community structure in fish-canning wastewater treatment plants.

Fish-canning wastewater is characterized frequently by a high content of salt (NaCl), making its treatment particularly difficult; however, the knowledge of the effect of NaCl on eukaryotic communities is very limited. In the present study, the global diversity of eukaryotes in activated sludges (AS) from 4 different wastewater treatment plants (WWTPs) treating fish-canning effluents varying in salinity (0.47, 1.36, 1.72 and 12.76 g NaCl/L) was determined by sequencing partial 18S rRNA genes using Illumina MiSeq. A greater diversity than previously reported was observed in the AS community, which comprised 37 and 330 phylum-like and genera-like groups, respectively. In this sense, the more abundant genus-like groups (average relative abundance (RA) > 5%) were Adineta (6.80%), Lecane (16.80%), Dictyostelium (7.36%), Unclassified_Fungi7 (6.94%), Procryptobia (5.13) and Oocystis (5.07%). The eukaryotic communities shared a common core of 25 phylum-like clades (95% of total sequences); therefore, a narrow selection of the eukaryotic populations was found, despite the differences in the abiotic characteristics of fish-canning effluents and reactor operational conditions inflicted. The differences in NaCl concentration were the main factor that influenced the structure of the eukaryotic community, modulating the RAs of the different phylum-like clades of the common core. Higher levels of salt increased the RAs of Ascomycota, Chlorophyta, Choanoflagellata, Cryptophyta, Mollusca, Nematoda, Other Protists and Unclassified Fungi. Among the different eukaryotic genera here found, the RA of Oocystis (Chlorophyta) was intimately correlated to increasing NaCl concentrations and it is proposed as a bioindicator of the global eukaryotic community of fish-canning WWTPs.

Evaluating the nitrogen-contaminated groundwater treatment by a denitrifying granular sludge bioreactor: effect of organic matter loading.

A sequential bed granular bioreactor was adapted to treat nitrate-polluted synthetic groundwater under anaerobic conditions and agitation with denitrification gas, achieving very efficient performance in total nitrogen removal at influent organic carbon concentrations of 1 g L-1 (80-90%) and 0.5 g L-1 (70-80%) sodium acetate, but concentrations below 0.5 g L-1 caused accumulation of nitrite and nitrate and led to system failure (30-40% removal). Biomass size and settling velocity were higher above 0.5 g L-1 sodium acetate. Trichosporonaceae dominated the fungal populations at all times, while a dominance of terrestrial group Thaumarchaeota and Acidovorax at 1 and 0.5 g L-1 passed to a domination of Methanobrevibacter and an unclassified Comamonadaceae clone for NaAc lower than 0.5 g L-1. The results obtained pointed out that the denitrifying granular sludge technology is a feasible solution for the treatment of nitrogen-contaminated groundwater, and that influent organic matter plays an important role on the conformation of microbial communities within it and, therefore, on the overall efficiency of the system.