Metabolism and cytotoxicity of diphenylarsinic acid, a degradation product of sea-dumped chemical warfare agents, in a rainbow trout liver cell line RTL-W1.

Recent studies have found primary degradation products of phenylarsenic chemical warfare agents (CWAs) accumulating in fish tissues, while the potential effects of these dumped phenylarsenic CWAs, such as Clark I and II, in the Baltic Sea biota are poorly understood. In this study, the metabolism and cytotoxicity of diphenylarsinic acid (DPA), a primary degradation product of phenylarsenic CWA, was studied by incubating rainbow trout cell line RTL-W1 cells in media with 100 mg/L DPA. Previously undescribed metabolites were identified by ultra-high performance liquid chromatography-high resolution mass spectrometry (UPHLCHRMS). Moreover, the cytotoxicity of diphenylarsine glutathione conjugate (DPA-SG), the major metabolite of DPA, was studied. Cytotoxicity of the compounds was evaluated using the Neutral Red retention test (NRR), showing an IC50 value of 278 mg/L for DPA and 1.30 mg/L for DPA-SG, indicating that the glutathione (GSH) conjugate of DPA is more than two orders of magnitude toxic than DPA itself, suggesting that toxic properties of DPA are increased after conjugation with intracellular GSH leading enhanced toxicity after uptake. Results gained in this study give more detailed information for elucidating biological effects of dumped chemical munitions in marine environment. Moreover, the results help in assessing the environmental and health risks posed by marine munition continued presence and deterioration in the sea bottom.

A preliminary study on the ecotoxic potency of wastewater treatment plant sludge combining passive sampling and bioassays.

Sewage sludge is an inevitable byproduct produced in wastewater treatment. Reusing nutrient-rich sludge will diminish the amount of waste ending in soil dumping areas and will promote circular economy. However, during sewage treatment process, several potentially harmful organic chemicals are retained in sludge, but proving the safety of processed sludge will promote its more extensive use in agriculture and landscaping. Environmental risk assessment of sludge requires new methods of characterizing its suitability for various circular economy applications. Bioavailable and bioaccessible fractions are key variables indicating leaching, transport, and bioaccumulation capacity. Also, sludge treatments have a significant effect on chemical status and resulting environmental risks. In this study, the concentrations of polyaromatic hydrocarbons (PAHs), triclosan (TCS), triclocarban (TCC), methyl triclosan (mTCS), and selected active pharmaceutical ingredients (APIs) were determined in different sludge treatments and fractions. Passive samplers were used to characterize the bioavailable and bioaccessible fractions, and the sampler extracts along the sludge and filtrate samples were utilized in the bioassays. The TCS and PAH concentrations did not decrease as the sludge was digested, but the contents diminished after composting. Also, mTCS concentration decreased after composting. The API concentrations were lower in digested sludge than in secondary sludge. Digested sludge was toxic for Aliivibrio fischeri, but after composting, toxicity was not observed. However, for Daphnia magna, passive sampler extracts of all sludge treatments were either acutely (immobility) or chronically (reproduction) toxic. Secondary and digested sludge sampler extracts were cytotoxic, and secondary sludge extract was also genotoxic. The measured chemical concentration levels did not explain the toxicity of the samples based on the reported toxicity thresholds. Bioassays and sampler extracts detecting bioavailable and bioaccessible contaminants in sludge are complementing tools for chemical analyses. Harmonization of these methodswill help establish scientifically sound regulative thresholds for the use of sludge in circular economy applications.

Effect-based assessment of toxicity removal during wastewater treatment.

Wastewaters contain complex mixtures of chemicals, which can cause adverse toxic effects in the receiving environment. In the present study, the toxicity removal during wastewater treatment at seven municipal wastewater treatment plants (WWTPs) was investigated using an effect-based approach. A battery of eight bioassays was applied comprising of cytotoxicity, genotoxicity, endocrine disruption and fish embryo toxicity assays. Human cell-based CALUX assays, transgenic larval models and the fish embryo toxicity test were particularly sensitive to WWTP effluents. The results indicate that most effects were significantly reduced or completely removed during wastewater treatment (76-100%), while embryo toxicity, estrogenic activity and thyroid disruption were still detectable in the effluents suggesting that some harmful substances remain after treatment. The responsiveness of the bioassays was compared and the human cell-based CALUX assays showed highest responsiveness in the samples. Additionally, the fish embryo toxicity test and the transgenic larval models for endocrine disrupting effects showed high responsiveness at low sample concentrations in nearly all of the effluent samples. The results showed a similar effect pattern among all WWTPs investigated, indicating that the wastewater composition could be rather similar at different locations. There were no considerable differences in the toxicity removal efficiencies of the treatment plants and no correlation was observed with WWTP characteristics, such as process configuration or sludge age. This study demonstrated that a biotest battery comprising of multiple endpoints can serve as a powerful tool when assessing water quality or water treatment efficiency in a holistic manner. Rather than analyzing the concentrations of a few selected chemicals, bioassays can be used to complement traditional methods of monitoring in the future by assessing sum-parameter based effects, such as mixture effects, and tackling chemicals that are present at concentrations below chemical analytical detection limits.

Previously uncultured beta-Proteobacteria dominate in biologically active granular activated carbon (BAC) filters.

Bacteria colonizing BAC filters used in drinking water purification from lake water were characterized by morphology, physiological tests, whole cell protein profiles and PLFA (phospholipid fatty acid) composition, and identified by partial 16S rRNA gene sequencing. Epifluorescence revealed prothecate bacteria to dominate in BAC. The majority of the isolates belonged to order Burkholderiales of beta-Proteobacteria, a few to Comamonadaceae but the majority to an undescribed family and the related sequences belonged mainly to uncultured bacteria. Among the less common alpha-Proteobacteria the genus Sphingomonas and the genera Afipia, Bosea or Bradyrhizobium of the Bradyrhizobiaceae family were detected. The majority of cultured bacteria persisting in the BAC biofilter were Burkholderiales, which according to ecological information are efficient in the mineralisation of dissolved organic matter in BAC. The biotechnical potential of the previously uncultured dominant bacteria warrants to be further studied.

Acetate and ethanol as potential enhancers of low temperature denitrification in soil contaminated by fur farms: a pilot-scale study.

Ethanol and acetate were examined as potential candidates to enhance denitrification at low temperature in soils contaminated by fur farms. Five pilot-scale sand and gravel columns with a top layer of soil from a fur farm were set up and fed with nitrate-containing water (influent concentration of 100 and 200 mg L(-1)) for 459 days at 6+/-2 degrees C. Two of the columns also received acetate and two other ethanol while one received no additional C-substrate. During the experiment, various C:N-ratios were tested to find the most optimal concentration of the added C-substrates, and effluent concentrations of nitrate, nitrite, and TOC were monitored. At the end of the experiments, soils in the columns were unpacked and the soils were used to measure a pattern of enzyme activities and the rates of denitrification in microcosms. The fur farm contaminated soil appeared to harbour a good intrinsic potential for denitrification, which could be greatly enhanced by the introduction of ethanol or acetate. Consequently, in the C-substrate-fed columns, 95-100% of the influent nitrate was removed after an acclimatization period of some weeks. Ethanol with C:N-ratio of ca. 6 at the nitrate level 200 mg L(-1) proved to be the most promising candidate to be used in field trials.