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).

Linking the shifts in the metabolically active microbiota in a UASB and hybrid anaerobic-aerobic bioreactor for swine wastewater treatment.

Due to the high concentration of pollutants, swine wastewater needs to be treated prior to disposal. The combination of anaerobic and aerobic technologies in one hybrid system allows to obtain higher removal efficiencies compared to those achieved via conventional biological treatment, and the performance of a hybrid system depends on the microbial community in the bioreactor. Here, we evaluated the community assembly of an anaerobic-aerobic hybrid reactor for swine wastewater treatment.

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.

Simplified engineering design towards a competitive lipid-rich effluents valorization.

Medium- and long-chain fatty acids and glycerol contained in the oily fraction of many food-industry effluents are excellent candidates to produce biobased high-value triacylglycerides (TAGs) and polyhydroxyalkanoates (PHAs). The typical process configuration for TAGs recovery from lipid-rich streams always includes two steps (culture enrichment plus storage compounds accumulation) whereas, for PHAs production, an additional pretreatment of the substrate for the obtainment of soluble volatile fatty acids (VFAs) is required. To simplify the process, substrate hydrolysis, culture enrichment, and accumulation (TAG and PHA storage) were coupled here in a single sequencing batch reactor (SBR) operated under the double growth limitation strategy (DGL) and fed in pulses with industrial waste fish oil during the whole feast phase.

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.

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.

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.

454-Pyrosequencing Analysis of Bacterial Communities from Autotrophic Nitrogen Removal Bioreactors Utilizing Universal Primers: Effect of Annealing Temperature.

Identification of anaerobic ammonium oxidizing (anammox) bacteria by molecular tools aimed at the evaluation of bacterial diversity in autotrophic nitrogen removal systems is limited by the difficulty to design universal primers for the Bacteria domain able to amplify the anammox 16S rRNA genes. A metagenomic analysis (pyrosequencing) of total bacterial diversity including anammox population in five autotrophic nitrogen removal technologies, two bench-scale models (MBR and Low Temperature CANON) and three full-scale bioreactors (anammox, CANON, and DEMON), was successfully carried out by optimization of primer selection and PCR conditions (annealing temperature). The universal primer 530F was identified as the best candidate for total bacteria and anammox bacteria diversity coverage.

Draft Genome Sequence of the Naphthalene Degrader Herbaspirillum sp. Strain RV1423.

Herbaspirillum sp. strain RV1423 was isolated from a site contaminated with alkanes and aromatic compounds and harbors the complete pathway for naphthalene degradation. The new features found in RV1423 increase considerably the versatility and the catabolic potential of a genus of bacteria previously considered mainly to be diazotrophic endophytes to plants.

Archaeal diversity in biofilm technologies applied to treat urban and industrial wastewater: recent advances and future prospects.

Biological wastewater treatment (WWT) frequently relies on biofilms for the removal of anthropogenic contaminants. The use of inert carrier materials to support biofilm development is often required, although under certain operating conditions microorganisms yield structures called granules, dense aggregates of self-immobilized cells with the characteristics of biofilms maintained in suspension. Molecular techniques have been successfully applied in recent years to identify the prokaryotic communities inhabiting biofilms in WWT plants.

Comparative analysis of the enzyme activities and the bacterial community structure based on the aeration source supplied to an MBR to treat urban wastewater.

A comparative analysis was performed in a pilot-scale membrane bioreactor (MBR) treating urban wastewater supplied with either pure oxygen (O2) or air, to assess the influence of each aeration source on the diversity and activity of the bacterial communities in the sludge. The MBR was operated in three experimental stages with different concentrations of volatile suspended solids (VSS) and temperature, and under both aeration conditions. α-Glucosidases, proteases, esterases and phosphatases were tested as markers of organic matter removal in the sludge, and the diversity of the bacterial community was analysed by fingerprinting (temperature-gradient gel electrophoresis of partially-amplified 16S-rRNA genes).

Bioremediation of diesel-polluted soil using biostimulation as post-treatment after oxidation with Fenton-like reagents: assays in a pilot plant.

The present study focuses on the remediation of diesel-polluted soil using modified Fenton treatment coupled with inorganic NPK fertilizer ("Fenton+NPK"). Studies were carried out in a pilot plant containing 1 m(3) of sandy soil contaminated with 20,000 mg kg(-1) of diesel, placed outdoors at a temperature ranging between 5 and 10 °C. Results showed that NPK-fertilizer as post-treatment stimulated culturable degrading bacteria and enhanced dehydrogenase activity.

Comparative analysis of the bacterial diversity in a lab-scale moving bed biofilm reactor (MBBR) applied to treat urban wastewater under different operational conditions.

Different types of carriers were tested as support material in a lab-scale moving bed biofilm reactor (MBBR) used to treat urban wastewater under three different conditions of hydraulic retention time (HRT) and carrier filling ratios (FR). The bacterial diversity developed on the biofilms responsible of the treatment was studied using a cultivation-independent approach based on the polymerase chain reaction-temperature gradient gel electrophoresis technique (PCR-TGGE). Cluster analysis of TGGE fingerprints showed significant differences of community structure dependent upon the different operational conditions applied.

Bacterial community structure and enzyme activities in a membrane bioreactor (MBR) using pure oxygen as an aeration source.

A pilot-scale membrane bioreactor was used to treat urban wastewater using pure oxygen instead of air as a source of aeration, to study its influence on bacterial diversity and levels of enzyme activities (acid and alkaline phosphatases, glucosidase, protease, and esterase) in the sludge. The experimental work was developed in two stages influenced by seasonal temperature. Operational parameters (temperature, pH, BOD5, COD, total and volatile suspended solids) were daily monitored, and enzyme activities measured twice a week.

Biofilm formation and microbial activity in a biofilter system in the presence of MTBE, ETBE and TAME.

Emerging water contaminants derived from unleaded gasoline such as methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE) and tert-amyl methyl ether (TAME), are in need of effective bioremediation technologies for restoring water resources. In order to design the conditions of a future groundwater bioremediating biofilter, this work assesses the potential use of Acinetobacter calcoaceticus M10, Rhodococcus ruber E10 and Gordonia amicalis T3 for the removal of MTBE, ETBE and TAME in consortia or as individual strains. Biofilm formation on an inert polyethylene support material was assessed with scanning electron microscopy, and consortia were also analysed with fluorescent in situ hybridisation to examine the relation between the strains.

Analysis of microbial communities developed on the fouling layers of a membrane-coupled anaerobic bioreactor applied to wastewater treatment.

The structure of the biofouling layers formed on a pilot-scale membrane-coupled upflow anaerobic sludge blanket bioreactor (UASB) used to treat urban wastewater was analyzed by scanning electron microscopy and electron-dispersive X-ray microanalysis. For comparison, control samples of the membranes were fed either UASB effluent or raw wastewater in a laboratory-scale experiment. Microbial diversity in the fouling materials was analyzed by temperature gradient gel electrophoresis (TGGE) combined with sequence analysis of partial 16S rRNA.

Metabolic characterization of a strain (BM90) of Delftia tsuruhatensis showing highly diversified capacity to degrade low molecular weight phenols.

A novel bacterium, strain BM90, previously isolated from Tyrrhenian Sea, was metabolically characterized testing its ability to use 95 different carbon sources by the Biolog system. The bacterium showed a broad capacity to use fatty-, organic- and amino-acids; on the contrary, its ability to use carbohydrates was extremely scarce. Strain BM90 was identified and affiliated to Delftia tsuruhatensis by molecular techniques based on 16S rRNA gene sequencing.

Influence of linear alkylbenzene sulfonate (LAS) on the structure of Alphaproteobacteria, Actinobacteria, and Acidobacteria communities in a soil microcosm.

Background, Aim, And Scope: Linear alkylbenzene sulfonate (LAS) is the most used anionic surfactant in a worldwide scale and is considered a high-priority pollutant. LAS is regarded as a readily biodegradable product under aerobic conditions in aqueous media and is mostly removed in wastewater treatment plants, but an important fraction (20-25%) is immobilized in sewage sludge and persists under anoxic conditions. Due to the application of the sludge as a fertilizer, LAS reaches agricultural soil, and therefore, microbial toxicity tests have been widely used to evaluate the influence of LAS on soil microbial ecology.

Effect of linear alkylbenzene sulfonates on the growth of aerobic heterotrophic cultivable bacteria isolated from an agricultural soil.

An enrichment culture technique was used to isolate soil bacteria capable of growing in the presence of two different concentrations of linear alkylbenzene sulfonates (LAS) (10 and 500 microg ml(-1)). Nine bacterial strains, representatives of the major colony types of aerobic heterotrophic cultivable bacteria in the enriched samples, were isolated and subsequently identified by PCR-amplification and partial sequencing of the 16S rRNA gene. Amongst the isolates, strains LAS05 (Pseudomonas syringae), LAS06 (Staphylococcus epidermidis), LAS07 (Delftia tsuruhatensis), LAS08 (Staphylococcus epidermidis) and LAS09 (Enterobacter aerogenes), were able to grow in pure culture in dialysed soil media amended with LAS (50 microg ml(-1)).

Production of chitinolytic enzymes by a strain (BM17) of Paenibacillus pabuli isolated from crab shells samples collected in the east sector of central Tyrrhenian Sea.

Nineteen bacterial isolates were grown in shaken cultures in media containing chitin as carbon source and different additional nitrogen sources such as yeast nitrogen base (YNB), yeast extract (YE), corn steep liquor (CSL) and ammonium sulfate. Strain BM17 showed the highest activity (200 U/l) in medium containing Chitin (1%) and YNB (0.5%).

Removal of organic load from olive washing water by an aerated submerged biofilter and profiling of the bacterial community involved in the process.

The present work aims to use a biofilter technology (aerated submerged filters) for the aerobic transformation at laboratory-scale of olive washing water (OWW) generated in the first steps of olive oil processing, as well as the genetic profiling and identification to the species level of the bacteria involved in the formation of the biofilm, by means of TGGE. Chemical parameters, such as biological oxygen demand at five days (BOD5) and chemical oxygen demand (COD), decreased markedly (up to 90 and 85%, respectively) by the biological treatment, and the efficiency of the process was significantly affected by aeration and inlet flow rates. The total polyphenol content of inlet OWW was only moderately reduced (around 50% decrease of the inlet content) after the biofilter treatment, under the conditions tested.

Growth and denitrifying activity of Xanthobacter autotrophicus CECT 7064 in the presence of selected pesticides.

The effects of the application of nine pesticides used commonly in agriculture (aldrin, lindane, dimetoate, methylparathion, methidation, atrazine, simazine, captan and diflubenzuron) on growth, CO2 production, denitrifying activity [as nitrous oxide (N2O) released] and nitrite accumulation in the culture medium by Xanthobacter autotrophicus strain CECT 7064 (Spanish Type Culture Collection) (a micro-organism isolated from a submerged fixed-film) were studied. The herbicide atrazine and the insecticide dimetoate totally inhibited growth and biological activity of X. autotrophicus at 10 mg l(-1), while the rest of the tested pesticides delayed the growth of strain CECT 7064 but did not drastically affect the bacterial growth after 96 h of culture.

Paenibacillus wynnii sp. nov., a novel species harbouring the nifH gene, isolated from Alexander Island, Antarctica.

Soil taken from 12 different locations at Mars Oasis on Alexander Island, Antarctica, yielded unidentified isolates of endospore-forming bacteria. Soil from four of the locations contained Gram-negative, facultatively anaerobic, motile rods that were able to grow at 4 degrees C and which formed ellipsoidal spores that lay paracentrally or subterminally in swollen or slightly swollen sporangia. All of the strains harboured the nitrogenase gene nifH.

Denitrifying activity of Xanthobacter autotrophicus strains isolated from a submerged fixed-film reactor.

Xanthobacter autotrophicus strains with the ability to reduce nitrate and nitrite to either nitrous oxide or molecular nitrogen were isolated from submerged fixed-film reactors. Isolated strains were Gram-negative rods able to grow on methanol, ethanol and sucrose. The yellow cellular pigmentation, pleomorphic appearance, and the presence of poly-beta-hydroxybutyrate granules suggest that the organisms might belong to the genus Xanthobacter.