Significantly differentially regulated biological processes in female <i>G. holbrooki</i> residing downstream of a paper mill as compared to a control site as determined by gene set enrichment and gene ontology (GO) analyses.

a<p> Percentage of genes within the GO category that were significantly differentially regulated between Econfina and Fenholloway groups.</p>b<p> Percentage of genes within the GO category that were not significantly differentially regulated between Econfina and Fenholloway groups.</p><p>Significantly differentially regulated biological processes in female <i>G. holbrooki</i> residing downstream of a paper mill as compared to a control site as determined by gene set enrichment and gene ontology (GO) analyses.</p

Gene expression profile comparisons between Fenholloway (PME) and reference site (Econ) female <i>G. holbrooki</i> and female <i>G. holbrooki</i> exposed to the androgen 17β-trenbolone (TB) or the vehicle control (C).

<p>This set of genes were differentially expressed between the PME, Econfina, and TB groups over the lab controls with at least a 1.5-fold difference of expression over the lab control and was expressed in the same direction in those groups. Data were median-centered by gene and clustered using spearman correlation and centroid linkage. Yellow genes are more highly expressed than the gene average and blue genes are expressed at a lower level than the gene average.</p

Focused hepatic gene expression patterns in female Eastern mosquitofish (<i>Gambusia holbrooki</i>) using qualitative polymerase chain reaction analysis.

<p>Gene expression of (A) vitellogenin (<i>vtg</i>), (B) zona pellucida glycoprotein 2 (<i>zp2</i>), (C) 17β-hydroxysteroid dehydrogenase 3 (<i>17βhsd3</i>) was analyzed between paper mill exposed (Fenholloway) and reference (Econfina) collection sites. Each point represents the mean of each group and error bars represent standard deviation; time points with no standard deviation have an N of 1. A box indicates statistical significance between groups at the selected oocyte developmental stages as determined using a Student's t-test with p<0.05.</p

Anal fin gene expression patterns of sonic hedgehog (<i>shh</i>) in female Eastern mosquitofish (<i>G. holbrooki</i>).

<p>An asterisk indicates statistical significance between the paper mill exposed (Fenholloway) and reference (Econfina) rivers as determined using a Student's t-test with p<0.05.</p

Anal fin elongation and bone segment formation in anal fin ray 3 of female Eastern mosquitofish (<i>Gambusia holbrooki</i>) collected from the Fenholloway and Econfina Rivers.

<p>Data are repsresented as (A) Mean (± standard deviation) of all collection events and, (B) Distribution of anal fin elongation classes between both paper mill exposed (Fenholloway) and reference (Econfina) field sites. An asterisk indicates statistical significance between the two groups as determined using a Student's t-test with p<0.05. For anal fin elongation levels there was an N of 46 and 50 from the Fenholloway and Econfina rivers respectively and a subset N of 4 from both sites for the bone segment evaluation.</p

Results of progesterone receptor (PR) and androgen receptor (AR) GeneBLAzer assays for concentrated water samples collected downstream of the Fenholloway River paper mill and the Econfina River conservation area.

<p>The graph bars represent the mean of three replicates from each dilution for the PR (A) and AR (B) assays. Error bars represent standard deviation of the three replicates per assay. The dose-responses of levonorgestrel, progesterone, and 17β-trenbolone (C) were also evaluated by the AR assay.</p

Silver Nanocolloids Disrupt Medaka Embryogenesis through Vital Gene Expressions

Silver nanomaterials are the major components of healthcare products largely because of their antimicrobial effects. However, their unintended toxicity to biological organisms and its mechanism are not well understood. Using medaka fish embryo model, the toxic effects and corresponding mechanisms of silver nanocolloids (SNC, particle size 3.8 ± 1.0-diameter nm) were investigated. SNC caused morphological changes in embryos including cardiovascular malformations, ischemia, underdeveloped central nervous system and eyes, and kyphosis at exposures of 0.5 mg/L. Interestingly, SNC were observed inside the eggs at a level of 786.1 ± 32.5 pg/mg egg weight, and TEM analysis showed that SNC adhered to the surface and inside of the chorion. Meanwhile, medaka oligo DNA microarray and qRT-PCR were used for gene expression analysis in the embryos exposed to 0.05 mg/L SNC for 48 h. As a result, expressions of six of the oxidative stress-, embryogenesis- and morphogenesis-related genes, <i>ctsL</i>, <i>tpm1</i>, <i>rbp</i>, <i>mt</i>, <i>atp2a1</i>, and <i>hox6b6</i>, were affected by the SNC exposure, and these genes’ involvement in those malformations was implied. Thus, SNC could potentially cause malformations in the cardiovascular and central nervous systems in developing medaka embryo through SNC-induced differential expression of the genes related to oxidative stress, embryonic cellular proliferation, and morphological development

Benchmarking Organic Micropollutants in Wastewater, Recycled Water and Drinking Water with In Vitro Bioassays

Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring