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Sulfamethoxazole triggers specific microbial activities under aerobic heterotrophic conditions: A metaproteomic approach

ABSTRACT: The global sanitary crisis derived from antibiotic multi-resistant bacteria entails the need to reduce sulfamethoxazole (SMX) concentrations in wastewater treatment plants (WWTPs). The key microorganisms and the biotransformation mechanisms leading to SMX removal remain incompletely characterized, particularly under aerobic heterotrophic conditions, which are becoming increasingly relevant in the design of novel, more energy-efficient, WWTPs. In this study, sequential batch reactors were inoculated with activated sludge, operated in heterotrophic conditions and spiked with six different initial SMX concentrations ranging between 0 and 2000 µg L-1. The goal was to determine the influence of SMX in the microbiome and its enzymatic expression through genomic, metaproteomic and transformation product analyses. The results allowed us to identify the metabolite 2,4(1H,3H)-pteridinedione-SMX (PtO-SMX), pointing to the role of the pterin-conjugation pathway in the biotransformation of SMX. Additionally, at increased SMX concentrations, through metaproteomics and 16S rRNA gene sequencing, it was determined a higher abundance of the genus Corynebacterium and a differential expression of five enzymes involved in its central metabolism, suggesting the relevant role of this bacteria to mitigate SMX risks.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Environmental Samples <g-proteobacteria Environmental Samples <b-proteobacteria

SUBMITTER: Valentina Calamia

LAB HEAD: Alba Trueba Santiso

PROVIDER: PXD029711 | Pride | 2022-09-20

REPOSITORIES: Pride

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Source:

			Action	DRS
	5-13-2021_SMX_Tf1000_100ng_1603.d.7z	Other
	5-13-2021_SMX_Tf1000_100ng_1603.mgf	Mgf
	5-13-2021_SMX_tf0_100ng_1587.d.7z	Other
	5-13-2021_SMX_tf0_100ng_1587.mgf	Mgf
	5-13-2021_SMX_tf250_100ng_1595.d.7z	Other

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Project description:The physiological and transcriptional response of Nitrosomonas europaea biofilms to phenol and toluene was examined and compared to suspended cells. Biofilms were grown in Drip Flow Biofilm Reactors under continuous flow conditions of growth medium containing ammonia as growth substrate. The responses of N. europaea biofilms to the aromatic hydrocarbons phenol and toluene were determined during short-term (3 h) additions of each compound to the biofilms. Ammonia oxidation in the biofilms was inhibited 50% by 60 uM phenol and 100 uM toluene. These concentrations were chosen for microarray analysis of phenol- and toluene-exposed N. europaea biofilms. Liquid batch cultures of exponentially growing N. europaea cells were harvested alongside the biofilms to determine differential gene expression between attached and suspended growth of N. europaea. Four sample groups of N. europaea cells were used in this study, with biological triplicates of each group. Groups were: Control (untreated) biofilms, phenol-exposed biofilms, toluene-exposed biofilms, and exponentially growing suspended cells. Biofilms were grown in Drip Flow Biofilm Reactors containing 4 independent growth channels and subject to 2 hour inhibition tests. During each experiment, 2 biofilm channels served as control with no inhibitor present and the other 2 biofilm channels were exposed to either 60 uM phenol or 100 uM toluene. Nitrite production was monitored throughout the experiment, and the given concentrations of phenol and toluene resulted in 50% inhibition of ammonia oxidation by the biofilms. Suspended cells were grown in batch reactors. Three 4-plex NimbleGen microarray chips were used, and each chip contained one sample from each experimental group. QC of samples was determined by spectrophotometric methods and using Agilent bioanalyzer traces to determine purity and integrity of RNA and cDNA. A sample tracking report was used to verify the correct hybridization of each sample to the intended array.

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Sulfamethoxazole triggers specific microbial activities under aerobic heterotrophic conditions: A metaproteomic approach

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