The Abundance and Diversity of Fungi in a Hypersaline Microbial Mat from Guerrero Negro, Baja California, México.

The abundance and diversity of fungi were evaluated in a hypersaline microbial mat from Guerrero Negro, México, using a combination of quantitative polymerase chain reaction (qPCR) amplification of domain-specific primers, and metagenomic sequencing. Seven different layers were analyzed in the mat (Layers 1-7) at single millimeter resolution (from the surface to 7 mm in depth). The number of copies of the 18S rRNA gene of fungi ranged between 106 and 107 copies per g mat, being two logarithmic units lower than of the 16S rRNA gene of bacteria. The abundance of 18S rRNA genes of fungi varied significantly among the layers with layers 2-5 mm from surface contained the highest numbers of copies. Fifty-six fungal taxa were identified by metagenomic sequencing, classified into three different phyla: Ascomycota, Basidiomycota and Microsporidia. The prevalent genera of fungi were Thermothelomyces, Pyricularia, Fusarium, Colletotrichum, Aspergillus, Botrytis, Candida and Neurospora. Genera of fungi identified in the mat were closely related to genera known to have saprotrophic and parasitic lifestyles, as well as genera related to human and plant pathogens and fungi able to perform denitrification. This research suggests that fungi in the mat may participate in nutrient recycling, modification of community composition through parasitic activities, and denitrification.

Assessment of the diversity and abundance of the total and active fungal population and its correlation with humification during two-phase olive mill waste (''alperujo") composting.

Metagenomic and transcriptomic techniques applied to composting could increase our understanding of the overall microbial ecology and could help us to optimise operational conditions which are directly related with economic interest. In this study, the fungal diversity and abundance of two-phase olive mill waste ("alperujo") composting was studied using Illumina MiSeq sequencing and quantitative PCR, respectively. The results showed an increase of the fungal diversity during the process, with Ascomycota being the predominant phylum. Penicillium was the main genera identified at the mesophilic and maturation phases, with Debaryomyces and Sarocladium at the thermophilic phase, respectively. The fungal abundance was increased during composting, which confirms their important role during thermophilic and maturation phases. Some Basidiomycota showed an increased during the process, which showed a positive correlation with the humification parameters. According to that, the genus Cystofilobasidium could be used as a potential fungal biomarker to assess alperujo compost maturation.

Evaluation of the Abundance of Fungi in Wastewater Treatment Plants Using Quantitative PCR (qPCR).

Assessment of the abundance of fungi in environmental samples by quantitative PCR (qPCR) of community DNA is often a difficult task due to biases introduced during PCR amplification, resulting from the differences associated with length polymorphism and the varying number of copies of the rRNA operon among fungal species, the lack of specificity of the primers targeting the different regions of the rRNA operon, or their insufficient coverage of the fungal lineages. To overcome those limitations, it is crucial to test and select the specific primers sets which provide the more accurate approximation to the quantification of the targeted fungal populations in a given set of samples. Fungi are a significant fraction of the microbiota in wastewater treatment plants (WWTPs), but the activated sludge microbial communities comprise many other eukaryotic microorganisms whose molecular markers are often coamplified by primers initially designed as fungal-specific. Here, the use of the FungiQuant primer set is recommended for the quantification of fungal molecular markers (18S rRNA genes) by qPCR in activated sludge samples and the full protocol is described.

Biodegradation of methyl and butylparaben by bacterial strains isolated from amended and non-amended agricultural soil. Identification, behavior and enzyme activities of microorganisms.

The aim of the present study was to investigate the kinetics of methylparaben (MPB) and butylparaben (BPB) removal, two emerging pollutants with possible endocrine disrupting effects, from agricultural soil with and without amendment with compost from sewage sludge used as biostimulant. Compound removal is explained by a first-order kinetic model with half-life times of 6.5/6.7 days and 11.4/8.2 days, in presence/absence of compost, for MPB and BPB respectively. % R2 for the fitted model were higher than 96% in all cases. Additionally, isolation of bacteria capable to grow using MPB or BPB as carbon source was also carry out. Laboratory tests demonstrated the ability of these bacteria to biodegrade MPB and BPB from culture media in more than 95% in some cases. These strains showed high ability to biodegrade the compounds. Ten isolates, most of them related to Gram positive bacteria of the genus Bacillus, were identified by 16S rRNA gene sequencing. The study of the enzymatic activities of the isolates revealed both esterase (C4) and esterase-lipase activities.

Microbial ecology dynamics of a partial nitritation bioreactor with Polar Arctic Circle activated sludge operating at low temperature.

A lab-scale partial nitritation SBR was operated at 11 °C for 300 days used for the treatment of high-ammonium wastewater, which was inoculated with activated sludge from Rovaniemi WWTP (located in Polar Arctic Circle) in order to evaluate the influence the temperature on the performance, stability and dynamics of its microbial community. The partial nitritation achieved steady-state long-term operation and granulation process was not affected despite the low temperature and high ammonia concentration. The steady conditions were reached after 60 days of operation where the granular biomass was fully-formed and the 50%-50% of ammonium-nitrite effluent was successful achieved. Inoculation with cold adapted inoculum showed to yield bigger, denser granules with faster start-up without necessity of low temperature adaptation period. Next-generation sequences techniques showed that Trichosporonaceae and Xanthomonadaceae were the dominant OTUs in the mature granules. Our study could be useful in the implementation of full-scale partial nitritation reactors in cold regions such as Nordic countries for treating wastewater with high concentration of ammonium.

Performance and microbial community structure of a polar Arctic Circle aerobic granular sludge system operating at low temperature.

The aim of this work was to study the performance and microbial community structure of a polar Arctic Circle aerobic granular sludge (AGS) system operating at low temperature. Thus, an AGS bioreactor was operated at 7, 5 and 3 °C of temperature using a cold-adapted sludge from Lapland. At 5 °C, it yielded acceptable conversion rates, in terms of nitrogen, phosphorous, and organic matter. However, under 3 °C a negligible nitrogen and phosphorous removal performance was observed. Below 5 °C, scanning electron microscopy studies showed a wispy, non-dense and irregular granular structure with a strong outgrowth of filamentous. Moreover, Illumina next-generation sequencing showed a heterogeneous microbial population where SM1K20 (Archaea), Trichosporon domesticum (Fungus), and Zooglea, Arcobacter and Acinetobacter (Bacteria) were the dominant phylotypes. Our study suggests that AGS technologies inoculated with North Pole sludge could be operated, in cold regions for a period longer than 3 months (winter season) under 5 °C of water temperature.

Nitrogen removal capacity and bacterial community dynamics of a Canon biofilter system at different organic matter concentrations.

Three Canon bench-scale bioreactors with a volume of 2 L operating in parallel were configured as submerged biofilters. In the present study we investigated the effects of a high ammonium concentration (320 mgNH4+· L-1) and different concentrations of organic matter (0, 100 and 400 mgCOD·L-1) on the nitrogen removal capacity and the bacterial community structure. After 60 days, the Canon biofilters operated properly under concentrations of 0 and 100 mgCOD·L-1 of organic matter, with nitrogen removal efficiencies up to 85%. However, a higher concentration of organic matter (400 mgCOD·L-1) produced a partial inhibition of nitrogen removal (68.1% efficiency). The addition of higher concentrations of organic matter a modified the bacterial community structure in the Canon biofilter, increasing the proliferation of heterotrophic bacteria related to the genera of Thauera, Longilinea, Ornatilinea, Thermomarinilinea, unclassified Chlorobiales and Denitratisoma. However, heterotrophic bacteria co-exist with Nitrosomonas and Candidatus Scalindua. Thus, our study confirms the co-existence of different microbial activities (AOB, Anammox and denitrification) and the adaptation of a fixed-biofilm system to different concentrations of organic matter.

Start-up and operation of an aerobic granular sludge system under low working temperature inoculated with cold-adapted activated sludge from Finland.

An aerobic granular sludge system has been started-up and operated at 7°C temperature using cold-adapted activated sludge as inoculum. The system could form granular biomass due to batch operation allowing for just 5-3min of biomass sedimentation. Scanning electron microscopy showed that fungi helped in the granular biomass formation in the early stages of the granule formation. The removal performance of the system was of 92-95% in BOD5, 75-80% in COD, 70-76% in total nitrogen and 50-60% in total phosphorous. The bacterial community structure from cold-adapted activated sludge changed during the operational time, leading to a final configuration dominated by Microbacteriaceae members Microbacterium and Leucobacter, which were strongly correlated to biomass settling velocity and bioreactor performance, as suggested by multivariate redundancy analyses. This experiment showed that aerobic granular sludge systems could be successfully started-up and operated, with high performance, under low operational temperatures when using cold-adapted biomass as inoculum.

Process performance and bacterial community dynamics of partial-nitritation biofilters subjected to different concentrations of cysteine amino acid.

Partial-nitritation processes are used for the biological treatment of high nitrogen-low organic carbon effluents, such as anaerobic digestion reject water. The release of certain products generated during the anaerobic digestion process, such as amino acids, could potentially reduce the performance of these partial-nitritation bioprocesses. To investigate this, four partial-nitritation biofilters were subjected to continuous addition of 0, 150, 300, and 500 mg L-1 cysteine amino acid in their influents. The addition of the amino acid had an impact over the performance of the partial-nitritation process and the bacterial community dynamics of the systems analyzed. Ammonium oxidation efficiency decreased with the addition of the amino acid, and a net nitrogen elimination occurred in presence of cysteine through the operation period. Bacterial community dynamics showed a decrease of Nitrosomonas species and a proliferation of putative heterotrophs with nitrification capacity, such as Pseudomonas, or denitrification capacity, such as Denitrobacter or Alicycliphilus. The addition of cysteine irreversible affected the bioreactors, which could not achieve the performance obtained before the addition of the amino acid. A mathematical predictive equation of the process performance depending on cysteine concentration added and operational time under such concentration was developed. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1254-1263, 2016.

Distribution and microbial community structure analysis of a single-stage partial nitritation/anammox granular sludge bioreactor operating at low temperature.

In the last decade, autotrophic nitrogen removal technologies based on anammox metabolism have become state of the art in urban and industrial wastewater treatment systems, due to their advantages over traditional nitrogen removal processes. However, their application is currently limited to the treatment of warm wastewater (25-40°C) mainly due to the low growth rate of the anammox bacteria. The extension of the application field to wastewater characterized by lower temperatures (8-20°C), such as those typical for municipal sewage, allows the design of treatment systems with a net energy production. In this study, the distribution and bacterial community structure of a lab-scale single-stage partial nitritation/anammox (PN/A) granular sludge bioreactor operating at low temperatures was analysed using next-generation sequencing techniques. The presence of ammonium-oxidizing bacteria and anammox bacteria was found, but the appearance of other bacterial species shows a complex microbial ecosystem. Evaluation of ecological roles of representative species inside the single-stage PN/A bioreactor was accomplished. Results obtained will be helpful for the future design and operation of PN/A systems performing at low temperatures.