Toxicokinetics and bioaccumulation of silver sulfide nanoparticles in benthic invertebrates in an indoor stream mesocosm.

Mesocosms allow the simulation of environmentally relevant conditions and can be used to establish more realistic scenarios of organism exposure to nanoparticles. An indoor mesocosm experiment simulating an aquatic stream ecosystem was conducted to assess the toxicokinetics and bioaccumulation of silver sulfide nanoparticles (Ag2S NPs) and AgNO3 in the freshwater invertebrates Girardia tigrina, Physa acuta and Chironomus riparius, and determine if previous single-species tests can predict bioaccumulation in the mesocosm. Water was daily spiked at 10 µg Ag L-1. Ag concentrations in water and sediment reached values of 13.4 µg Ag L-1 and 0.30 µg Ag g-1 in the Ag2S NP exposure, and 12.8 µg Ag L-1 and 0.20 µg Ag g-1 in the AgNO3. Silver was bioaccumulated by the species from both treatments, but with approximately 1.5, 3 and 11 times higher body Ag concentrations in AgNO3 compared to Ag2S NP exposures in snails, chironomids and planarians, respectively. In the Ag2S NP exposures, the observed uptake was probably of the particulate form. This demonstrates that this more environmentally relevant Ag nanoform may be bioavailable for uptake by benthic organisms. Interspecies interactions likely occurred, namely predation (planarians fed on chironomids and snails), which somehow influenced Ag uptake/bioaccumulation, possibly by altering organisms´ foraging behaviour. Higher Ag uptake rate constants were determined for AgNO3 (0.64, 80.4 and 1.12 Lwater g-1organism day-1) than for Ag2S NPs (0.05, 2.65 and 0.32 Lwater g-1organism day-1) for planarians, snails and chironomids, respectively. Biomagnification under environmentally realistic exposure seemed to be low, although it was likely to occur in the food chain P. acuta to G. tigrina exposed to AgNO3. Single-species tests generally could not reliably predict Ag bioaccumulation in the more complex mesocosm scenario. This study provides methodologies/data to better understand exposure, toxicokinetics and bioaccumulation of Ag in complex systems, reinforcing the need to use mesocosm studies to improve the risk assessment of environmental contaminants, specifically NPs, in aquatic environments.

Toxicokinetics of Ag from Ag2S NP exposure in Tenebrio molitor and Porcellio scaber: Comparing single-species tests to indoor mesocosm experiments.

Determining the potential for accumulation of Ag from Ag2S NPs as an environmentally relevant form of AgNPs in different terrestrial organisms is an essential component of a realistic risk assessment of AgNP emissions to soils. The objectives of this study were first to determine the uptake kinetics of Ag in mealworms (Tenebrio molitor) and woodlice (Porcellio scaber) exposed to Ag2S NPs in a mesocosm test, and second, to check if the obtained toxicokinetics could be predicted by single-species bioaccumulation tests. In the mesocosms, mealworms and woodlice were exposed together with plants and earthworms in soil columns spiked with 10 µg Ag g-1 dry soil as Ag2S NPs or AgNO3. The total Ag concentrations in the biota were measured after 7, 14, and 28 days of exposure. A one-compartment model was used to calculate the Ag uptake and elimination rate constants. Ag from Ag2S NPs appeared to be taken up by the mealworms with significantly different uptake rate constants in the mesocosm compared to single-species tests (K1 = 0.056 and 1.66 g dry soil g-1 dry body weight day-1, respectively), and a significant difference was found for the Ag bioaccumulation factor (BAFk = 0.79 and 0.15 g dry soil g-1 dry body weight, respectively). Woodlice did not accumulate Ag from Ag2S NPs in both tests, but uptake from AgNO3 was significantly slower in mesocosm than in single-species tests (K1 = 0.037 and 0.26 g dry soil g-1 dry body weight day-1, respectively). Our results are of high significance because they show that single-species tests may not be a good predictor for the Ag uptake in mealworms and woodlice in exposure systems having greater levels of biological complexity. Nevertheless, single-species tests could be used as a fast screening approach to assess the potential of a substance to accumulate in biota before more complex tests are conducted.

In vitro models to detect in vivo bile acid changes induced by antibiotics.

Bile acid homeostasis plays an important role in many biological activities through the bile-liver-gut axis. In this study, two in vitro models were applied to further elucidate the mode of action underlying reported in vivo bile acid changes induced by antibiotics (colistin sulfate, tobramycin, meropenem trihydrate, and doripenem hydrate). 16S rRNA analysis of rat fecal samples anaerobically incubated with these antibiotics showed that especially tobramycin induced changes in the gut microbiota. Furthermore, tobramycin was shown to inhibit the microbial deconjugation of taurocholic acid (TCA) and the transport of TCA over an in vitro Caco-2 cell layer used as a model to mimic intestinal bile acid reuptake. The effects induced by the antibiotics in the in vitro model systems provide novel and complementary insight explaining the effects of the antibiotics on microbiota and fecal bile acid levels upon 28-day in vivo treatment of rats. In particular, our results provide insight in the mode(s) of action underlying the increased levels of TCA in the feces upon tobramycin exposure. Altogether, the results of the present study provide a proof-of-principle on how in vitro models can be used to elucidate in vivo effects on bile acid homeostasis, and to obtain insight in the mode(s) of action underlying the effect of an antibiotic, in this case tobramycin, on bile acid homeostasis via effects on intestinal bile acid metabolism and reuptake.

Metal transfer to sediments, invertebrates and fish following waterborne exposure to silver nitrate or silver sulfide nanoparticles in an indoor stream mesocosm.

The fate of engineered nanomaterials in ecosystems is unclear. An aquatic stream mesocosm explored the fate and bioaccumulation of silver sulfide nanoparticles (Ag2S NPs) compared to silver nitrate (AgNO3). The aims were to determine the total Ag in water, sediment and biota, and to evaluate the bioavailable fractions of silver in the sediment using a serial extraction method. The total Ag in the water column from a nominal daily dose of 10 µg L-1 of Ag for the AgNO3 or Ag2S NP treatments reached a plateau of around 13 and 12 µg L-1, respectively, by the end of the study. Similarly, the sediment of both Ag-treatments reached ~380 µg Ag kg-1, and with most of it being acid-extractable/labile. The biota accumulated 4-59 µg Ag g-1 dw, depending on the type of Ag-treatment and organism. The oligochaete worm, Lumbriculus variegatus, accumulated Ag from the Ag2S exposure over time, which was similar to the AgNO3 treatment by the end of the experiment. The planarian, Girardia tigrina, and the chironomid larva, Chironomus riparius, showed much higher Ag concentrations than the oligochaete worms; and with a clearer time-dependent statistically significant Ag accumulation relative to the untreated controls. For the pulmonate snail, Physa acuta, bioaccumulation of Ag from AgNO3 and Ag2S NP exposures was observed, but was lower from the nano treatment. The AgNO3 exposure caused appreciable Ag accumulation in the water flea, Daphnia magna, but accumulation was higher in the Ag2S NP treatment (reaching 59 µg g-1 dw). In the rainbow trout, Oncorhynchus mykiss, AgNO3, but not Ag2S NPs, caused total Ag concentrations to increase in the tissues. Overall, the study showed transfer of total Ag from the water column to the sediment, and Ag bioaccumulation in the biota, with Ag from Ag2S NP exposure generally being less bioavailable than that from AgNO3.

Are long-term exposure studies needed? Short-term toxicokinetic model predicts the uptake of metal nanoparticles in earthworms after nine months.

Uptake of most metal nanoparticles (NPs) in organisms is assumed to be mainly driven by the bioavailability of the released ions, as has been verified in controlled and short-term exposure tests. However, the changeability of NPs and the dynamic processes which NPs undergo in the soil environment, bring uncertainty regarding their interactions with soil organisms over a long period of time. To assess the potential impacts of long-term exposure scenarios on the toxicokinetic of metal NPs, earthworms Eisenia fetida were exposed to soils spiked with pristine Ag-NP, aged Ag-NP (Ag2S-NP) and ionic Ag for nine months, and results were compared to those from a similar short-term (28 days) experiment, conducted under similar conditions. Overall, there were no statistical differences between long-term accumulation patterns in earthworms exposed to pristine Ag-NP and AgNO3, while for Ag2S-NP, the amount of Ag internalized after 9 months was five times lower than for the other treatments. Average Ag concentrations in soil pore water in all treatments did not change over time, however the soil pH decreased and electrical conductivity increased in all treatments. Metallothionein concentrations in exposed earthworms were not statistically different from levels in untreated earthworms. Finally, the short-term toxicokinetic models predicted the bioaccumulation in earthworms exposed to Ag-NP, AgNO3 after nine months on the whole. Although the bioaccumulation for Ag2S-NPs was somewhat under-predicted, the rate of accumulation of Ag2S-NPs is much lower than that of Ag-NPs or AgNO3 and thus potentially of lower concern. Nevertheless, better understanding about the exposure kinetics of Ag2S-NP would help to address potential nano-specific toxicokinetic and toxicodynamics, also of other sulfidized metal NPs.

Species Differences in in vitro and Estimated in vivo Kinetics for Intestinal Microbiota Mediated Metabolism of Acetyl-deoxynivalenols.

SCOPE: Deoxynivalenol (DON) and its acetylated derivatives 3-acetyl-DON (3-Ac-DON) and 15-acetyl-DON (15-Ac-DON) are important mycotoxins of concern in the modern food chain. METHODS AND RESULTS: The present study reveals that the rate of de-acetylation in in vitro anaerobic fecal incubations decreased in the order rat > mouse > human > pig for 3-Ac-DON, and mouse > human > rat > pig for 15-Ac-DON. The ratio between the de-acetylation rate of 3-Ac-DON and 15-Ac-DON varies with the species. Scaling of the kinetic parameters to the in vivo situation results in catalytic efficiencies decreasing in the order human > rat > pig > mouse for 3-Ac-DON and human > pig > rat > mouse for 15-Ac-DON. The results obtained indicate that in mice, 3-Ac-DON can be fully deconjugated while 15-Ac-DON cannot. In rats, pigs, and humans, both 3-Ac-DON and 15-Ac-DON can be totally transformed by gut fecal microbiota during the estimated intestinal residence time. A correlation analysis between the deacetylation rate and the relative abundance of the microbiome suggests Lachnospiraceae may be involved in the deacetylation process. CONCLUSION: It is concluded that interspecies differences in deacetylation of acetylated DONs exist but that in risk assessment assumption of complete intestinal deconjugation provides an adequate approach.

Interindividual Differences in Human Intestinal Microbial Conversion of (-)-Epicatechin to Bioactive Phenolic Compounds.

To quantify interindividual differences in the human intestinal microbial metabolism of (-)-epicatechin (EC), in vitro anaerobic incubations with fecal inocula from 24 healthy donors were conducted. EC-derived colonic microbial metabolites were qualitatively and quantitively analyzed by liquid chromatography triple quadrupole mass spectrometry (LC-TQ-MS) and liquid chromatography time-of-flight mass spectrometry (LC-TOF-MS). Quantitative microbiota characterization was achieved by 16S rRNA analysis. The results obtained show 1-(3',4'-dihydroxyphenyl)-3-(2″,4″,6″-dihydroxyphenyl)-2-propanol (3,4-diHPP-2-ol) and 5-(3',4'-dihydroxyphenyl)-γ-valerolactone (3,4-diHPV) to be key intermediate microbial metabolites of EC and also revealed the substantial interindividual differences in both the rate of EC conversion and the time-dependent EC metabolite pattern. Furthermore, substantial differences in microbiota composition among different individuals were detected. Correlations between specific microbial phylotypes and formation of certain metabolites were established. It is concluded that interindividual differences in the intestinal microbial metabolism of EC may contribute to interindividual differences in potential health effects of EC-abundant dietary foods or drinks.

Bioturbation of Ag2S-NPs in soil columns by earthworms.

Sewage sludge contains Ag2S-NPs causing NP exposure of soil fauna when sludge is applied as soil amendment. Earthworm bioturbation is an important process affecting many soil functions. Bioturbation may be affected by the presence of Ag2S-NPs, but the earthworm activity itself may also influence the displacement of these NPs that otherwise show little transport in the soil. The aim of this study was to determine effects of Ag2S-NPs on earthworm bioturbation and effect of this bioturbation on the vertical distribution of Ag2S-NPs. Columns (12 cm) of a sandy loamy soil with and without Lumbricus rubellus were prepared with and without 10 mg Ag kg-1, applied as Ag2S-NPs in the top 2 cm of the soil, while artificial rainwater was applied at ∼1.2 mm day-1. The soil columns were sampled at three depths weekly for 28 days and leachate collected from the bottom. Total Ag measurements showed more displacement of Ag to deeper soil layers in the columns with earthworms. The application of rain only did not significantly affect Ag transport in the soil. No Ag was detected in column leachates. X-ray tomography showed that changes in macro porosity and pore size distribution as a result of bioturbation were not different between columns with and without Ag2S-NPs. Earthworm activity was therefore not affected by Ag2S-NPs at the used exposure concentration. Ag concentrations along the columns and the earthworm density allowed the calculation of the bioturbation rate. The effect on the Ag transport in the soil shows that earthworm burrowing activity is a relevant process that must be taken into account when studying the fate of nanoparticles in soils.