Metabolism of the14C-labeled herbicide clodinafop-propargyl in plant cell cultures of wheat and tobacco

The metabolism of ¹⁴C-clodinafop-propargyl (CfP) was examined in cell cultures of wheat (Triticum aestivum L. cv. ‘Heines Koga II’) and tobacco (Nicotiana tabacum L.). Besides the non-transgenic tobacco culture, cultures transformed separately with cDNA of human cytochrome P450-monooxygenases (P450s) CYP1A1, CYP1A2, CYP3A4, CYP2B6 and CYP2C19 were examined. Experiments with wheat were executed in the presence and absence of safener cloquintocet-mexyl (CqM). After 48 h of incubation, only about 10% of applied ¹⁴C was found in media (both tobacco and wheat). Non-extractable residues of ¹⁴C-CfP in wheat cells were 16.54% (without CqM) and 30.87% (with CqM). In all tobacco cultures, 82.41–92.46% of applied radioactivity was recovered in cell extracts. In contrast to wheat, non-extractable residues amounted only to 1.50–2.82%. As determined by radio-thin layer chromatography (TLC) and -high-performance liquid chromatography (HPLC), the parent CfP was not found in the cell extracts of wheat; in tobacco cell extracts, only traces of CfP were detected. After a hydrolysis of assumed carbohydrate conjugates of CfP derived polar ¹⁴C-labeled compounds, TLC and HPLC analysis showed that in wheat, a more complex pattern of metabolites of CfP were observed as compared to all tobacco cultures. In hydrolysates resulting from wheat, the identity of three primary products was confirmed by means of GC-EI-MS: free acid clodinafop (Cf), hydroxy-Cf hydroxylated at the pyridinyl moiety, and 4-(5-chloro-3-fluoropyridin-2-yloxy)phenol. In hydrolysates derived from all tobacco cultures, main metabolite was Cf besides only traces of further unidentified products. Differences among the different tobacco cultures (non-transgenic, transgenic) did not emerge. According to kinetics of disappearance of primary metabolite Cf as well as formation of polar soluble products and non-extractable residues, metabolization of CfP proceeded at a noticeably higher rate in wheat cells treated with safener CqM than in cells without CqM treatment. Thus, these results indicated a stimulation of CfP's metabolism by CqM, although metabolic profiles observed in CqM treated and non-treated cells (after hydrolysis) were qualitatively similar. The findings obtained from all tobacco cultures suggested that with the exception of ester cleavage to Cf, CfP cannot be metabolized by tobacco itself or by the human P450s examined.     

Multiresidue analysis of organophosphate and pyrethroid pesticides in duplicate-diet solid food by pressurized liquid extraction

An analytical method was developed for determining organophosphate pesticides (OPP) and pyrethroid pesticides (PYR) in duplicate-diet solid food. The method consisted of pressurized liquid extraction (PLE) with dichloromethane followed by cleanup with gel permeation and solid phase extraction columns and gas chromatography/mass spectrometry (GC/MS) analysis. Quantitative recoveries (73–117 %) of the target pesticides were obtained for spiked duplicate-diet food samples. The percent standard deviation (% RSD) of replicate food samples was within ± 20 %. Another method was developed for determining a common OPP metabolite, 3, 5, 6-trichloro-2-pyridinol (TCP) in duplicate-diet food. The method consisted of a PLE with methanol followed by liquid-liquid partitioning, derivatization, and GC/MS analysis. Recoveries of TCP ranged from 83 to 101 % for spiked duplicate-diet food samples. The % RSD of replicate food samples was within ± 15 %. The results confirmed that these methods are reliable and robust, and that they can be used in routine analysis. In addition, a storage stability study for a common OPP, chlorpyrifos (CPF), in solid food samples was performed. The fortified ¹⁵N-¹³C-labeled CPF was stable over 16 mo storage at ?20° C in the dark. The developed analytical methods were successfully applied to 278 duplicate-diet food samples from preschool children, demonstrating that these methods are robust and suitable for routine analysis in future exposure monitoring studies.     

Using radon-222 as indicator for the evaluation of the efficiency of groundwater remediation by in situ air sparging

A common approach for remediation of groundwater contamination with volatile organic compounds (VOCs) is contaminant stripping by means of in situ air sparging (IAS). For VOC stripping, pressurized air is injected into the contaminated groundwater volume, followed by the extraction of the contaminant-loaded exhaust gas from the vadose soil zone and its immediate on-site treatment. Progress assessment of such remediation measure necessitates information (i) on the spatial range of the IAS influence and (ii) on temporal variations of the IAS efficiency. In the present study it was shown that the naturally occurring noble gas radon can be used as suitable environmental tracer for achieving the related spatial and temporal information. Due to the distinct water/air partitioning behaviour of radon and due to its straightforward on-site detectability, the radon distribution pattern in the groundwater can be used as appropriate measure for assessing the progression of an IAS measure as a function of space and time. The presented paper discusses both the theoretical background of the approach and the results of an IAS treatment accomplished at a VOC contaminated site lasting six months, during which radon was applied as efficiency indicator.     

Distribution, fate and formation of non-extractable residues of a nonylphenol isomer in soil with special emphasis on soil derived organo-clay complexes

Anthropogenic contaminants like nonylphenols (NP) are added to soil, for instance if sewage-sludge is used as fertilizer in agriculture. A commercial mixture of NP consists of more than 20 isomers. For our study, we used one of the predominate isomers of NP mixtures, 4-(3,5-dimethylhept-3-yl)phenol, as a representative compound. The aim was to investigate the fate and distribution of the isomer within soil and soil derived organo-clay complexes. Therefore, (14)C- and (13)C-labeled NP was added to soil samples and incubated up to 180 days. Mineralization was measured and soil samples were fractionated into sand, silt and clay; the clay fraction was further separated in humic acids, fulvic acids and humin. The organo-clay complexes pre-incubated for 90 or 180 days were re-incubated with fresh soil for 180 days, to study the potential of re-mobilization of incorporated residues. The predominate incorporation sites of the nonylphenol isomer in soil were the organo-clay complexes. After 180 days of incubation, 22 % of the applied (14)C was mineralized. The bioavailable, water extractable portion was low (9 % of applied (14)C) and remained constant during the entire incubation period, which could be explained by an incorporation/release equilibrium. Separation of organo-clay complexes, after extraction with solvents to release weakly incorporated, bioaccessible portions, showed that non-extractable residues (NER) were preferentially located in the humic acid fraction, which was regarded as an effect of the chemical composition of this fraction. Generally, 27 % of applied (14)C was incorporated into organo-clay complexes as NER, whereas 9 % of applied (14)C was bioaccessible after 180 days of incubation. The re-mobilization experiments showed on the one hand, a decrease of the bioavailability of the nonylphenol residues due to stronger incorporation, when the pre-incubation period was increased from 90 to 180 days. On the other hand, a shift of these residues from the clay fraction to other soil fractions was observed, implying a dynamic behavior of incorporated residues, which may result in bioaccessibility of the NER of nonylphenol.     

Genetic diversity and IUCN Red List status

The International Union for Conservation of Nature (IUCN) Red List is an important and widely used tool for conservation assessment. The IUCN uses information about a species’ range, population size, habitat quality and fragmentation levels, and trends in abundance to assess extinction risk. Genetic diversity is not considered, although it affects extinction risk. Declining populations are more strongly affected by genetic drift and higher rates of inbreeding, which can reduce the efficiency of selection, lead to fitness declines, and hinder species’ capacities to adapt to environmental change. Given the importance of conserving genetic diversity, attempts have been made to find relationships between red-list status and genetic diversity. Yet, there is still no consensus on whether genetic diversity is captured by the current IUCN Red List categories in a way that is informative for conservation. To assess the predictive power of correlations between genetic diversity and IUCN Red List status in vertebrates, we synthesized previous work and reanalyzed data sets based on 3 types of genetic data: mitochondrial DNA, microsatellites, and whole genomes. Consistent with previous work, species with higher extinction risk status tended to have lower genetic diversity for all marker types, but these relationships were weak and varied across taxa. Regardless of marker type, genetic diversity did not accurately identify threatened species for any taxonomic group. Our results indicate that red-list status is not a useful metric for informing species-specific decisions about the protection of genetic diversity and that genetic data cannot be used to identify threat status in the absence of demographic data. Thus, there is a need to develop and assess metrics specifically designed to assess genetic diversity and inform conservation policy, including policies recently adopted by the UN's Convention on Biological Diversity Kunming-Montreal Global Biodiversity Framework.     

Fungal biotransformation of 6:2 fluorotelomer alcohol

Fungal degradation of 6:2 fluorotelomer alcohol (6:2 FTOH, C₆F₁₃CH₂CH₂OH) by two wood‐decaying fungal strains and six fungal isolates from a site contaminated with per‐ and polyfluoroalkyl substances (PFASs) was investigated. 6:2 FTOH is increasingly being used in FTOH‐based products, and previous reports on the microbial fate of 6:2 FTOH have focused on bacteria and environmental microbial consortia. Prior to this study, one report demonstrated that the 6:2 FTOH biotransformation by the wood‐decaying fungus, Phanerochaete chrysosporium, generated more polyfluoroalkyl substances, such as 5:3 acid (F(CF₂)₅CH₂CH₂COOH), and diverted away from producing the highly stable perfluorocarboxylic acids (PFCAs). Most of the fungi (Gloeophyllum trabeum and isolates TW4‐2, TW4‐1, B79, and B76) examined in this study showed similar degradation patterns, further demonstrating that fungi yield more 5:3 acid (up to 51 mol% of initial 6:2 FTOH dosed) relative to other metabolites (up to 12 mol% total PFCAs). However, medium amendments can potentially improve 6:2 FTOH biotransformation rates and product profiles. The six fungal isolates tolerated up to 100 or 1,000 milligrams per liter of perfluorooctanoic acid and perfluorooctane sulfonic acid, and some isolates experienced increased growth with increasing concentrations. This study proposes that fungal pathways must be considered for the biotransformation of potential PFAS precursors, such as 6:2 FTOH, and suggests the basis for selecting proper microorganisms for remediation of fluoroalkyl‐contaminated sites.     

Predicting microalgae growth and phosphorus removal in cold region waste stabilization ponds using a stochastic modelling approach

A stochastic ecological model with an integrated equilibrium temperature model was developed to predict microalgae growth and phosphorus removal in cold region waste stabilization ponds (WSPs). The model utilized a Monte Carlo simulation to account for parameter uncertainty. The equilibrium temperature model was parameterized using field data collected from two WSPs in Nunavut, Canada, from 2012 to 2014. The equilibrium temperature model provided good agreement with field data on a daily time step. The full model was run using historic (1956–2005) temperature and solar radiation data from five communities (Baker Lake, Cambridge Bay, Coral Harbour, Hall Beach, Resolute) in Nunavut, Canada. The communities represented a range of geographical locations and environmental conditions. Logistic regression on pooled model outputs showed that mean July temperature and mean treatment season temperature (June 1–September 15, ice-free period) provided the best predictors for microalgae growth. They had a predictive success rate of 93 and 88%, respectively. The modelled threshold (50% probability from the Monte Carlo simulation) for microalgae growth was 8.7 and 5.6 °C for the July temperature and mean treatment season temperature, respectively. The logistic regression was applied to each community (except Sanikiluaq) in Nunavut using historic climate data and a probability of microalgae growth was calculated. Based on the model results, soluble phosphorus concentrations consistent with secondary treatment could be achieved if WSP depth is less than 2 m. The model demonstrated a robust method to predict whether a microalgae bloom will occur under a range of model parameters.     

Impact of Dredging on Phosphorus Transport in Agricultural Drainage Ditches of the Atlantic Coastal Plain1

Abstract: Drainage ditches can be a key conduit of phosphorus (P) between agricultural soils of the Atlantic Coastal Plain and local surface waters, including the Chesapeake Bay. This study sought to quantify the effect of a common ditch management practice, sediment dredging, on fate of P in drainage ditches. Sediments from two drainage ditches that had been monitored for seven years and had similar characteristics (flow, P loadings, sediment properties) were sampled (0‐5 cm) after one of the ditches had been dredged, which removed fine textured sediments (clay = 41%) with high organic matter content (85 g/kg) and exposed coarse textured sediments (clay = 15%) with low organic matter content (2.2 g/kg). Sediments were subjected to a three‐phase experiment (equilibrium, uptake, and release) in recirculating 10‐m‐long, 0.2‐m‐wide, and 5‐cm‐deep flumes to evaluate their role as sources and sinks of P. Under conditions of low initial P concentrations in flume water, sediments from the dredged ditch released 13 times less P to the water than did sediments from the ditch that had not been dredged, equivalent to 24 mg dissolved P. However, the sediments from the dredged ditch removed 19% less P (76 mg) from the flume water when it was spiked with dissolved P to approximate long‐term runoff concentrations. Irradiation of sediments to destroy microorganisms revealed that biological processes accounted for up to 30% of P uptake in the coarse textured sediments of the dredged ditch and 18% in the fine textured sediments of the undredged ditch. Results indicate that dredging of coastal plain drainage ditches can potentially impact the P buffering capacity of ditches draining agricultural soils with a high potential for P runoff.