Assessment of cellular and molecular metrics for dose selection in an in vivo comet assay: A case study with MDI.

The in vivo comet assay can evaluate the genotoxic potential of a chemical in theoretically any tissue that can be processed to a single cell suspension. This flexibility enables evaluation of point-of-contact tissues using a relevant route of test material administration; however, assessing cytotoxicity is essential for the interpretation of comet results. Histopathological evaluation is routinely utilized to assess cytotoxicity, but temporal- and cell-specific considerations may compromise applicability to the comet assay. In the present study, 1,1'-methylenebis(4-isocyanatobenzene) (4,4'-MDI) was administered to rats for 6 h by nose-only inhalation, and the comet assay was conducted to evaluate genotoxicity in the site-of-contact tissue (bronchoalveolar lavage cells) and distal tissues (liver and glandular stomach). Given the reactive nature of MDI, cellular and molecular metrics at the site-of-contact- including inflammation, macrophage activation, apoptosis/necrosis, and oxidative stress- were used to set appropriate exposure concentrations, in addition to the standard systemic measures of toxicity. In the range-finding study, a concentration of 4 mg/m3 was considered the maximum noninflammatory concentration; hence target concentrations of 2, 5, and 11 mg/m3 were selected for the comet study. In the lung lavage, MDI exposure substantially increased total protein and ß-glucuronidase, along with cellular apoptosis. Although MDI did not increase the comet assay response (% tail DNA) in any of the tissues examined, the positive control (ethyl methanesulfonate, EMS) significantly increased % tail DNA in all tissues. In total, these data indicate that appropriate cellular and molecular measurements may facilitate dose selection to discern cellular status in the comet assay.

Monitoring water quality in Toronto's urban stormwater ponds: Assessing participation rates and data quality of water sampling by citizen scientists in the FreshWater Watch.

From 2013 to 2015, citizen scientist volunteers in Toronto, Canada were trained to collect and analyze water quality in urban stormwater ponds. This volunteer sampling was part of the research program, FreshWater Watch (FWW), which aimed to standardize urban water sampling efforts from around the globe. We held training sessions for new volunteers twice yearly and trained a total of 111 volunteers. Over the course of project, ~30% of volunteers participated by collecting water quality data after the training session with 124 individual sampling events at 29 unique locations in Toronto, Canada. A few highly engaged volunteers were most active, with 50% of the samples collected by 5% of trainees. Stormwater ponds generally have poor water quality demonstrated by elevated phosphate concentrations (~30µg/L), nitrate (~427µg/L), and turbidity relative to Canadian water quality standards. Compared to other urban waterbodies in the global program, nutrient concentrations in Toronto's urban stormwater ponds were lower, while turbidity was not markedly different. Toronto FWW (FWW-TO) data was comparable to that measured by standard lab analyses and matched results from previous studies of stormwater ponds in Toronto. Combining observational and chemical data acquired by citizen scientists, macrophyte dominated ponds had lower phosphate concentrations while phytoplankton dominated ponds had lower nitrate concentrations, which indicates a potentially important and unstudied role of internal biogeochemical processes on pond nutrient dynamics. This experience in the FWW demonstrates the capabilities and constraints of citizen science when applied to water quality sampling. While analytical limits on in-field analyses produce higher uncertainty in water quality measurements of individual sites, rapid data collection is possible but depends on the motivation and engagement of the group of volunteers. Ongoing efforts in citizen science will thus need to address sampling effort and analytical limits to fully realize the potential value of engaging citizen scientists in water quality sampling.

Chronic and pulse exposure effects of silver nanoparticles on natural lake phytoplankton and zooplankton.

The increasing use of silver nanoparticles (AgNPs) in consumer products raises concerns regarding the environmental exposure and impact of AgNPs on natural aquatic environments. Here, we investigated the effects of environmentally relevant AgNP concentrations on the natural plankton communities using in situ enclosures. Using twelve lake enclosures, we tested the hypotheses that AgNP concentration, dosing regimen, and capping agent (poly-vinyl pyrrolidone (PVP) vs. citrate) exhibit differential effects on plankton communities. Each of the following six treatments was replicated twice: control (no AgNPs added), low, medium, and high chronic PVP treatments (PVP-capped AgNPs added continuously, with target nominal concentrations of 4, 16, and 64 µg/L, respectively), citrate treatment (citrate-capped AgNPs added continuously, target nominal concentrations of 64 µg/L), and pulse treatment (64 µg/L PVP-AgNPs added as a single dose). Although Ag accumulated in the phytoplankton, no statistically significant treatment effect was found on phytoplankton community structure or biomass. In contrast, as AgNP exposure rate increased, zooplankton abundance generally increased while biomass and species richness declined. We also observed a shift in the size structure of zooplankton communities in the chronic AgNP treatments. In the pulse treatments, zooplankton abundance and biomass were reduced suggesting short periods of high AgNP concentrations affect zooplankton communities differently than chronic exposures. We found no evidence that capping agent affected AgNP toxicity on either community. Overall, our study demonstrates variable AgNP toxicity between trophic levels with stronger AgNP effects on zooplankton. Such effects on zooplankton are troubling and indicate that AgNP contamination could affect aquatic food webs.