ABSTRACT: Analysis of SNP variation in candidate genes to identify genes associated with adaptation to exposure to wastewater treatment work effluents.
Project description:Many biomonitoring tools/approaches have been proposed to assess presence of endocrine active chemicals (EACs) and their biological effects in the field. Although these tools have provided valuable information, they are often limited by their specificity for certain groups of EACs and they may not account for interactions between EACs. This study aims to evaluate utility of transcriptomic and metabolomic technologies for effects monitoring in the field, and to advance integration of omic and environmental chemistry data sets. The objective was to utilize transcriptomic biomonitoring to determine the relative contribution of wastewater treatment plant effluents to biological effects observed in fish exposed to ambient waters receiving the effluents.
Project description:Many biomonitoring tools/approaches have been proposed to assess presence of endocrine active chemicals (EACs) and their biological effects in the field. Although these tools have provided valuable information, they are often limited by their specificity for certain groups of EACs and they may not account for interactions between EACs. This study aims to evaluate utility of transcriptomic and metabolomic technologies for effects monitoring in the field, and to advance integration of omic and environmental chemistry data sets. The objective was to utilize transcriptomic biomonitoring to determine the relative contribution of wastewater treatment plant effluents to biological effects observed in fish exposed to ambient waters receiving the effluents. Adult male fathead minnow were exposed to treated wastewater effluent or stream water up or downstream the plant in three different watersheds for 4 days. After exposure, the liver of 5-7 fish per treatment per site (i.e 19-21 fish from each watershed) were analyzed by microarrays. The transcriptomic profiles were compared to control fish exposed to Lake Superior filtered water.
Project description:The transcriptome analysis by the human DNA microarray was applied to evaluate the impacts of whole wastewater effluents from the membrane bioreactors (MBRs) and the activated sludge process (AS), on the biological processes of human hepatoma HepG2 cells. The three conventional bioassays (i.e., cytotoxicity tests and bioluminescence inhibition test) and chemical analysis of the domestic effluent standards were conducted in parallel since they are well-established methods with previous applications to wastewater. A significant variation of effluent quality was sdemonstrated among the tested effluents despite that all effluents met the 40 national effluent standards. The three conventional bioassays supported the result of the transcriptome analysis, indicating the comparable or even higher sensitivity of the new assay. The most superior effluent quality was found in the MBR operated at a relatively long sludge retention time (i.e., 40 days) and small membrane pore size (i.e., 0.03 M-NM-<m). In addition, functional analysis of the differentially expressed genes revealed that the effluents made various impacts on the cellular functions, suggesting the transcriptome analysis by DNA microarray as more comprehensive, rapid and sensitive tool to detect multiple impacts of the whole effluents. Moreover, the potential genetic markers were proposed to quantitatively evaluate the treatability of the wastewater effluents. In this study, we examined the gene expression alteration in human hepatoma cell line, HepG2 exposed to the raw wastewater, effluents from three types of membrane bioreactors (MBRs), and the activated sludge process. Wastewater DNA microarray with 8795 human genes. MQ water was used as control. For duplicate, two dishes were prepared for each sample and individually treated in parallel.
Project description:The transcriptome analysis by the human DNA microarray was applied to evaluate the impacts of whole wastewater effluents from the membrane bioreactors (MBRs) and the activated sludge process (AS), on the biological processes of human hepatoma HepG2 cells. The three conventional bioassays (i.e., cytotoxicity tests and bioluminescence inhibition test) and chemical analysis of the domestic effluent standards were conducted in parallel since they are well-established methods with previous applications to wastewater. A significant variation of effluent quality was sdemonstrated among the tested effluents despite that all effluents met the 40 national effluent standards. The three conventional bioassays supported the result of the transcriptome analysis, indicating the comparable or even higher sensitivity of the new assay. The most superior effluent quality was found in the MBR operated at a relatively long sludge retention time (i.e., 40 days) and small membrane pore size (i.e., 0.03 μm). In addition, functional analysis of the differentially expressed genes revealed that the effluents made various impacts on the cellular functions, suggesting the transcriptome analysis by DNA microarray as more comprehensive, rapid and sensitive tool to detect multiple impacts of the whole effluents. Moreover, the potential genetic markers were proposed to quantitatively evaluate the treatability of the wastewater effluents.
Project description:The increase in human population and urbanization are resulting in an increase in the volume of wastewater and urban runoff effluents entering natural ecosystems. These effluents may contain multiple pollutants to which the biological response of aquatic organisms is still poorly understood mainly due to mixture toxicity and interactions with other environmental factors. In this context, RNA sequencing was used to assess the impact of a chronic exposure to wastewater treatment plant and stormwater effluents at the whole-transcriptome level and evaluate the potential physiological outcomes in the Asian clam Corbicula fluminea. We de-novo assembled a transcriptome from C. fluminea digestive gland and identified a set of 3,181 transcripts with altered abundance in response to water quality. The largest differences in transcriptomic profiles were observed between C. fluminea from the reference site and those exposed to wastewater treatment plant effluents. On both anthropogenically impacted sites, most differentially expressed transcripts were involved in signaling pathways in relation to energy metabolism such as mTOR and FoxO, suggesting an energy/nutrient deficit and hypoxic conditions. These conditions were likely responsible for damages to proteins and transcripts in response to wastewater treatment effluents whereas exposure to urban runoff might result in immune and endocrine disruptions. In absence of comprehensive chemical characterization, the RNAseq approach could provide information regarding the mode of action of pollutants and then be useful for the identification of which parameters must be studied at higher integration level in order to diagnose sites where the presence of complex and variable mixtures of chemicals is suspected.
Project description:Laboratory tests with marine flatfish were conducted to investigate associations among gene expression, higher biological responses and wastewater effluent exposure. Previous studies showed molecular responses such as elevated concentrations of plasma estradiol and vitellogenin in wild male hornyhead turbot (Pleuronichthys verticalis). In the present study, male hornyhead turbot were exposed to environmentally realistic (0.5%) and higher (5%) concentrations of chemically enhanced advanced-primary (PL) and full-secondary treated (HTP) effluents from two southern California wastewater treatment plants (WWTP). Hepatic gene expression was examined using a custom low-density microarray. <br><br>
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.
Project description:Three surface waters in Gainesville, Florida were used in a 48 hour whole effluents exposure to assess gene expression profiles of male fathead minnow liver. Microarray analysis was used to determine changes in gene expression of exposed fish to waters from a site downstream of a wastewater treatment plant (streamwater), a wastewater treatment plant (wastewater), and a lake (stormwater). Differences in gene expression between fish exposed to collected waters and controls were observed. Number of altered genes and biological processes were 1028 and 18 for stormwater; 787 and 19 for streamwater; and: 575 and 12 for wastewater. In general, the effects observed in all exposed fish were related with fatty acid metabolism, DNA repair, oxidation-reduction process, cell wall catabolic process and apoptosis. All exposed fish showed altered expression of genes related with DNA damage repair. In particular fish exposed to stormwater and streamwater showed downregulation of several key intermediates transcripts of cholesterol. The presence and environmental persistence of perfluorinated chemicals (PFCs) in these waters, the resemblance in known effects on transcripts with those found in this study, suggest that the set of genes differentially regulated in fathead minnows after 48 hours of exposure may be attributed to exposure to PFCs.
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.
Project description:<p>Due to a unique adaptation to high altitude, the Tibetan Plateau population has been the subject of much research interest. In this study, we conducted whole genome sequencing of 27 Tibetan individuals. Through our analysis, we inferred a detailed history of demography and revealed the natural selection of Tibetan population. We provided evidence of genetic separation between the two subpopulations of Han and Tibetans as early as 44 to 58 thousand years ago, replicated previously reported high altitude adaptation genes, including <i>EPAS1</i> and <i>EGLN1</i>, and reported three new candidate genes, including <i>PTGIS</i>, <i>VDR</i>, and <i>KCTD12</i>.</p>