Project description:Despite recent knowledge of the potential environmental impact that compounds present in municipal wastewater effluents, including contaminants of emerging concern (CECs), may have, the implications of fish exposure to this contaminant mixtures are not completely understood. The effects caused by effluent CECs may be subtle and diverse, thus the need for sensitive and comprehensive tools such as gene expression to detect such responses. In this study, we conducted laboratory exposures that examined plasma concentrations of vitellogenin (VTG), changes in secondary sexual characteristics and gene expression in sexually mature male fathead minnows (Pimephales promelas) exposed to environmentally realistic (0.5%) and higher (5%) concentrations of municipal wastewater effluents. Secondary and primary treated effluents were used. Several of the 32 CECs investigated were detected, including pharmaceuticals, personal care products, hormones, current use pesticides and industrial compounds. The percent of males with detectable levels of VTG was higher in fish exposed to effluent treatments. An increased number of males with changes in secondary sexual characteristics (e.g. development of ovipositors), was observed in fish exposed to 5% effluent treatments. Gene expression data indicated that overall expression patterns were characteristic to each effluent. Higher numbers of differentially expressed genes were observed in fish exposed to primary treated effluent when compared to controls. Differentially expressed genes belonged to several functional categories, including xenobiotic metabolism, estogenicity and energy/metabolism processes. Gene expression data provided information to understand some of the mechanisms behind the effects observed at higher biological levels. To investigate gene expression responses resulting from exposure to POTW effluents, two laboratory experiments were conducted using effluent from San Diego (Point Loma; SD) and Los Angeles (Hyperion; LA). The LA effluent received secondary treatment and the SD effluent received advanced primary treatment. Treatments used during exposures consisted of negative controls (moderately hard water), positive controls (E2), and 0.5% and 5% effluent concentrations. The 0.5% concentration of effluent represented an environmentally realistic exposure level. The 5% effluent concentration represented a higher level at which we expected biological responses. The exposures lasted 14 days. Treatments: EFFHa = 5% primary treated effluent EFFHb = 5% secondary treated effluent EFFLa = 0.5% primary treated effluent E2a = Estradiol, positive control for primary effluent E2b = Estradiol, positive control for secondary effluent CTRLa = Moderately hard water, negative control for primary effluent CTRLb = Moderately hard water, negative control for secondary effluent
Project description:Contaminants of emerging concern (CECs) in treated municipal effluents have the potential to adversely impact exposed organisms prompting elevated public concern. Using transcriptomic tools, we investigated changes in gene expression and cellular pathways in the liver of male fathead minnows (Pimephales promelas) exposed to 5% concentrations of full secondary-treated (HTP) or advanced primary-treated (PL) municipal wastewater effluents containing CECs. Gene expression changes were associated with apical endpoints (plasma vitellogenin and changes in secondary sexual characteristics). Of 32 effluent CECs analyzed, 28 were detected including pharmaceuticals, personal care products, hormones, and industrial compounds. Transcript patterns differed between effluents, however < 10% of these had agreement in the detected response (e.g. transcrips involved in xenobiotic detoxification, oxidative stress and apoptosis) in both effluents. Exposure to PL effluent caused changes in transcript levels of genes involved in metabolic pathways (e.g., lipid transport and steroid metabolism). Exposure to HTP effluent affected transcripts involved in signaling pathways (e.g., focal adhesion assembly and extracellular matrix). Exposure to both effluents produced significantly higher levels of plasma VTG and changes in secondary sexual characteristics (e.g., ovipositor development). Taken together the results suggest, a potential association between some transcriptomic changes and higher biological responses following effluent exposure; and a potential adverse outcome pathway following exposure to complex chemical mixtures containing CECs -. Furthermore, this study identified responses in key genes and pathways not previously implicated in exposure to CECS, , which could be consistent with effluent exposure (e.g., oxidative stress) in addition to other pathway responses specific to the effluent type. This may be useful for assessing the adverse health effects of fish by effluents exposure to CECs.
2011-05-18 | GSE29350 | GEO
Project description:Microvolume DNA Extraction Methods for Microscale Amplicon and Metagenomic Studies
Project description:The following were run:
Blanks
Solvent blanks (with and without NIS/EIS)
Extraction blanks (with NIS/EIS)
Wastewater Extracts with NIS/EIS
Fish Creek Final Effluent 220902
Fish Creek Final Effluent 220903
Bonnybrook Final Effluent 220902
Bonnybrook Final Effluent 220902 - Duplicate
Pine Creek Final Effluent 220902
Pine Creek Final Effluent 220902 - Duplicate
Standards
EPA 1633 standards
EPA 1633 NIS
EPA 1633 EIS
Calibration curve of standards with EIS & NIS
Wastewater Extract Pools
Fish Creek Pooled
Bonnybrook Pooled
Pine Creek Pooled
The purpose was to apply FluoroMatch and NTA to these samples as well as compare QRAI, AIF, and IE-DDA using Innovative Omics new deconvolution tool: IonDecon
Project description:Effect of chlorination on the toxicity of wastewater effluents treated by activated sludge (AS) and submerged membrane bioreactor (S-MBRB) systems to HepG2 human hepatoblastoma cells was investigated. In addition to cytotoxicity assay, the DNA microarray-based transcriptome analysis was performed to evaluate the change in modes of toxic actions (MOAs) of effluents by chlorination. Effluent organic matters (EfOM) and disinfection by-products (DBPs) were characterized by using Fourier transform mass spectrometry (FT-MS). The cytotoxicity of AS effluent was elevated by chlorination, while the toxicity of S-MBRB effluent was reduced. The averaged O/C ratio of EfOM in S-MBRB effluent was lower than that in AS effluent. The results of the transcriptome and FT-MS analyses suggested that lower O/C molecules influenced on M-bM-^@M-^\response to hormone stimulusM-bM-^@M-^] and M-bM-^@M-^\acute inflammatory responseM-bM-^@M-^] but those were decreased by chlorination, which consequently reduced cytotoxicity. On the other hand, larger number of DBPs and other molecules were increased in AS effluents by chlorination. Those molecules might influence on M-bM-^@M-^\cellular metabolic processM-bM-^@M-^], which consequently elevated cytotoxicity. Therefore, the combination of the toxicity assays and chemical analysis demonstrated the changes in severity of cytotoxicity and MOAs by chlorination, and the difference of chemical characteristics which relate to those toxicity changes. We examined the gene expression alteration in human hepatoma cell line, HepG2 exposed to the chlorinated wastewater effluents from membrane bioreactor and the activated sludge process. Human Genome Focus Array, which represents 8,795 verified human sequences, was used. All effluent samples were concentrated by using solid phase extraction (SPE). SPE fraction from MQ water was used as controll. For duplicate, two dishes were prepared for each sample and individually treated in parallel.