Nontarget screening using passive air and water sampling with a level II fugacity model to identify unregulated environmental contaminants.

It is thought that there are many unregulated anthropogenic chemicals in the environment. For risk assessment of chemicals, it is essential to estimate the predicted environmental concentrations. As an effort of identifying residual organic contaminants in air and water in Korea, nontarget screening using two-dimensional gas chromatography time-of-flight mass spectrometry (GC×GC-TOFMS) was conducted at 10 sites using polyurethane foam passive air sampler and at 6 sites using polydimethyl siloxane (PDMS) passive water sampler in three different seasons in 2014. More than 600 chemical peaks were identified satisfying the identification criteria in air and water samples, respectively, providing a list for further investigation. Chemical substances with reported national emission rates in 2014 (n=149) were also screened for potential existence in the environment using a level II fugacity model. Most of chemical substances classified as not detectable were not identified with detection frequency greater than 20% by nontarget screening, indicating that a simple equilibrium model has a strong potential to be used to exclude chemicals that are not likely to remain in the environment after emissions from targeted monitoring.

Internal Concentration and Time Are Important Modifiers of Toxicity: The Case of Chlorpyrifos on Caenorhabditis elegans.

The internal concentration of chemicals in exposed organisms changes over time due to absorption, distribution, metabolism, and excretion processes since chemicals are taken up from the environment. Internal concentration and time are very important modifiers of toxicity when biomarkers are used to evaluate the potential hazards and risks of environmental pollutants. In this study, the responses of molecular biomarkers, and the fate of chemicals in the body, were comprehensively investigated to determine cause-and-effect relationships over time. Chlorpyrifos (CP) was selected as a model chemical, and Caenorhabditis elegans was exposed to CP for 4 h using the passive dosing method. Worms were then monitored in fresh medium during a 48-h recovery regime. The mRNA expression of genes related to CYP metabolism (cyp35a2 and cyp35a3) increased during the constant exposure phase. The body residue of CP decreased once it reached a peak level during the early stage of exposure, indicating that the initial uptake of CP rapidly induced biotransformation with the synthesis of new CYP metabolic proteins. The residual chlorpyrifos-oxon concentration, an acetylcholinesterase (AChE) inhibitor, continuously increased even after the recovery regime started. These delayed toxicokinetics seem to be important for the extension of AChE inhibition for up to 9 h after the start of the recovery regime. Comprehensive investigation into the molecular initiation events and changes in the internal concentrations of chemical species provide insight into response causality within the framework of an adverse outcome pathway.

Strong links between load and manure and a comprehensive risk assessment of veterinary antibiotics with low KOW in intensive livestock farming watersheds.

Various veterinary antibiotics (VAs) are used in large quantities as an essential component for intensive livestock farming, and can flow into the environment from various pollution sources. In this study, VAs in surface water and groundwater in the Gwangcheon stream watershed, an intensive livestock farming area in Korea, were analyzed using ultra-high-performance liquid chromatograph-quadrupole orbitrap high-resolution mass spectrometer with online solid phase extraction. Although the selected VAs are relatively mobile and have low KOW values it is significant to assess their fates and ecological risks in the environment. The concentration of VAs in the surface water was higher than that in groundwater by approximately 23-fold, indicating that the former were directly introduced from pollution sources such as livestock manure. An analysis of the correlation between livestock manure production and the residuals of VAs in the stream showed a high linearity (R2 > 0.70), confirming that livestock excreta significantly contributed to the VAs in the watershed. A combined evaluation of environmental behaviors and ecological risks of VAs was performed for the first time using persistence, bioaccumulation potential, and toxicity properties and risk quotient values of VAs. Trimethoprim showed persistence and a potential impact on the ecosystem. The cumulative risk quotient values at one sampling point exceeded 1 indicating that several VAs can cumulatively cause local risk. The risk assessment method considering pollution sources, different locations, and correlation analysis applied in this study will be useful in evaluating the impacts of trace pollutants in watersheds.

Concentrations and Risk Assessments of Antibiotics in an Urban-Rural Complex Watershed with Intensive Livestock Farming.

Antibiotics used for the treatment of humans and livestock are released into the environment, whereby they pose a grave threat to biota (including humans) as they can cause the emergence of various strains of resistant bacteria. An improved understanding of antibiotics in the environment is thus vital for appropriate management and mitigation. Herein, surface water and groundwater samples containing antibiotics were analyzed in an urban-rural complex watershed (Cheongmi Stream) comprising intensive livestock farms by collecting samples across different time points and locations. The spatiotemporal trends of the residual antibiotics were analyzed, and ecological and antibiotic resistance-based risk assessments were performed considering their concentrations. The results showed that the concentrations and detection frequencies of the residual antibiotics in the surface water were affected by various factors such as agricultural activities and point sources, and were higher than those found in groundwater; however, frequent detection of antibiotics in groundwater showed that residual antibiotics were influenced by factors such as usage pattern and sewage runoff. Furthermore, few antibiotics posed ecological risks. The risk assessment methods adopted in this study can be applied elsewhere, and the results can be considered in the environmental management of residual antibiotics in the Cheongmi Stream watershed.

Identifying nitrogen sources in intensive livestock farming watershed with swine excreta treatment facility using dual ammonium (δ15NNH4) and nitrate (δ15NNO3) nitrogen isotope ratios axes.

We proposed a novel approach based on dual ammonium and nitrate nitrogen isotope ratios (δ15NNH4 and δ15NNO3, respectively) axes to identify nitrogen sources in intensive livestock farming watersheds, especially those with swine excreta treatment facilities. The δ15NNH4 and δ15NNO3 values in water samples were measured monthly in 2016-2017. Soil and mineral fertilizers, sewage, sewage effluent, manure, and swine effluents were the five sources considered to identify nitrogen sources. The results showed that nitrogen pollution from agricultural activities was well reflected by the seasonal δ15NNH4 and δ15NNO3 patterns in the river, and microbial nitrification was suggested as the dominant nitrogen transformation process in the river. This study revealed that δ15NNH4 and δ15NNO3 axes provided better results than the traditionally used nitrate oxygen (δ18ONO3) and δ15NNO3 axes for identifying nitrogen sources in agricultural watersheds with swine excreta treatment facilities. The mixing model results showed that stream water was severely contaminated with swine effluents (e.g., a mean minimum contribution of 31%), thus affecting the quality of the mainstream (p = 0.068 < 0.10). This study was the first successful application of dual δ15NNH4 and δ15NNO3 axes to better understand nitrogen sources in intensive livestock farming watersheds with swine excreta treatment facilities.

Bioavailability of hydrophobic organic chemicals on an in vitro metabolic transformation using rat liver S9 fraction.

Metabolic transformation of highly hydrophobic organic chemicals (HOCs) is one of the most important factors modulating their persistence, bioaccumulation and toxicity. Although sorption of HOCs to cellular matrices affects their bioavailability, it is still not clear how the cellular binding or sorption of HOCs in in vitro metabolism assays influences their enzymatic transformation kinetics. To elucidate effects of non-specific binding to enzymes, we measured apparent enzyme kinetics in an in vitro assay using four polycyclic aromatic hydrocarbons (phenanthrene, anthracene, pyrene and benzo[a]pyrene) as model HOCs and S9 mixture isolated from rat liver as a model enzyme mixture. The effects were also investigated in the presence of bovine serum albumin (BSA), which served to isolate the effect of protein binding from transformation. The observed transformation rates were much higher than those predicted assuming that only freely dissolved HOCs are available for metabolism. A new model including kinetic exchanges between non-specifically bound HOCs and those bound to active enzyme binding sites explained the apparent transformation kinetics at various experimental conditions better. The results are relevant for in vitro-in vivo extrapolation because the metabolic transformation rate in vivo may depend strongly on the local enzyme density and the micro-cellular environment. While non-specific protein binding reduces the unbound fraction of chemicals, this effect could be partially compensated by the facilitated transport to the active sites of the enzymes.

Characteristics of veterinary antibiotics in intensive livestock farming watersheds with different liquid manure application programs using UHPLC-q-orbitrap HRMS combined with on-line SPE.

Composted livestock manures, in both solid and liquid form, are used as fertilizers in cropland. However, excess solid and liquid manures in agricultural watersheds are considered as nonpoint pollution sources because of their high nutrient and heavy metal contents of, as well as their antibiotic contents, especially veterinary antibiotics (VAs). In this study, 21 VAs under nine classes (i.e., cephems, ionophores, lincosamides, penicillins, pleuromutilins, quinolones, streptogramins, sulfonamides, and tetracyclines) found in agricultural watersheds were simultaneously analyzed via UHPLC-q-orbitrap high-resolution mass spectrometry using an on-line solid-phase extraction system. The residues of VAs in the surface water of two intensive livestock rearing watersheds (Cheongmi and Gwangcheon streams) in Korea were successfully quantified, and the values were found to range from 1.84 ± 0.42 ng L-1 to 835.6 ± 31.9 ng L-1. Time lags of 2-3 months were observed between the periods of liquid manure application and the periods with the maximum concentrations of VAs. In both watersheds, samples from points close to areas with extensive application of liquid manure exhibited high concentrations of most of the 21 VAs. Between the watersheds, the one with heavier application of liquid manure showed higher concentrations of the target VAs. To the best of our knowledge, this study represents the first attempt at evaluating the correlation between liquid manure application and environmental occurrence of VAs in surface water. The findings reveal that liquid manure application plays an important role in introducing VAs into aquatic environments.

Estimating microplastic-bound intake of hydrophobic organic chemicals by fish using measured desorption rates to artificial gut fluid.

One of the most important concerns about marine microplastics is their role in delivery of chemical contaminants to biota. The contribution of microplastic ingestion to the overall uptake of five hydrophobic organic chemicals (HOCs) [α-, ß-, and γ-hexachlorocyclohexanes (HCHs), pentachlorobenzene (PeCB), and hexachlorobenzene (HeCB)] by fish is evaluated in this study. Partition coefficients of all five HOCs between surfactant micelles and simulated intestinal fluid (SIF), as well as between protein and SIF, were experimentally determined. Desorption of model HOCs from a polyethylene film into an artificial gut solution was measured to estimate the fraction of HOCs that can be absorbed from microplastics during their gut retention time. Monte-Carlo simulation (n = 100,000) showed that the uptake via microplastic ingestion will be negligible for HCHs as compared to uptake via other exposure routes, water ventilation and food ingestion. On the other hand, microplastic ingestion might increase the total uptake rate of PeCB and HeCB due to their accelerated desorption from microplastics into the artificial gut solution under the model scenario, assuming an extremely high intake of microplastics. However, the steady-state bioaccumulation factor was predicted to decrease with increasing ingestion of microplastics, showing a dilution effect by microplastic ingestion. Results indicate that HOCs that are close to be at phase equilibrium between microplastics and environmental media are not likely to be further accumulated via ingestion of microplastics; this is true even for cases, where ingestion of microplastics contributes significantly to the total uptake of HOCs. Therefore, future studies need to focus on hydrophobic plastic additives that may exist in microplastics at a concentration higher than their equilibrium concentration with water.

Evaluation of the bioaccumulation potential of selected alternative brominated flame retardants in marine fish using in vitro metabolic transformation rates.

The global consumption of alternative brominated flame retardants (BFRs) has increased with the restriction of the first generation BFRs such as polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecanes (HBCDs). However, many alternative BFRs are suspected to be persistent in the environment and possibly bioaccumulative after their release into the environment because of their chemical properties, which are similar to those of the already banned BFRs. In this study, the bioaccumulation potential of selected alternative BFRs (1,2-bis(2,4,6­tribromophenoxy)ethane (BTBPE), 1,2,3,4,5,6­hexabromobenzene (HBB), pentabromoethylbenzene (PBEB), 2,3,4,5,6­pentabromotoluene (PBT), 2­ethylhexyl-2,3,4,5-tetrabromobenzoate (TBB), and 2,3,4,5­tetrabromo-6-chlorotoluene (TBCT)) was evaluated. The in vitro depletion rate constants (kdepl) were measured for the alternative BFRs using liver S9 fractions isolated from five marine fish species (Epinephelus septemfasciatus, Konosirus punctatus, Lateolabrax japonicus, Mugil cephalus, and Sebastes schlegelii) that inhabit the oceans off the Korean coast. The measured kdepl values were converted to in vitro intrinsic clearance rate constants (CLin vitro) to estimate whole-body metabolic rate constants (kMET) using an in vitro to in vivo extrapolation (IVIVE) model. Finally, the bioconcentration factors (BCF) were determined using a one-compartment model. The transformation kinetics for obtaining kdepl agreed well with first-order chemical kinetics, regardless of initial BFR concentrations. The values of CLin vitro were lower in the selected marine fish species than those in freshwater fish species, implying slower metabolic transformation. The derived BCF values based on the total concentration in water (BCFTOT) ranged from 16 (TBB in M. cephalus) to 27,000 (HBB in K. punctatus). The BCF values for HBB and PBT were >2000 except for those in M. cephalus suggesting further investigation of BCF values of BFRs whose log KOW is between 6 and 7.

Erratum to: Epilepsy and Other Neuropsychiatric Manifestations in Children and Adolescents with 22q11.2 Deletion Syndrome.

This corrects the article on p. 85 in vol. 12, PMID: 26754781.

Epilepsy and Other Neuropsychiatric Manifestations in Children and Adolescents with 22q11.2 Deletion Syndrome.

BACKGROUND AND PURPOSE: 22q11.2 deletion syndrome (22q11.2DS) is the most common microdeletion syndrome. Epilepsy and other neuropsychiatric (NP) manifestations of this genetic syndrome are not uncommon, but they are also not well-understood. We sought to identify the characteristics of epilepsy and other associated NP manifestations in patients with 22q11.2DS. METHODS: We retrospectively analyzed the medical records of 145 child and adolescent patients (72 males and 73 females) with genetically diagnosed 22q11.2DS. The clinical data included seizures, growth chart, psychological reports, development characteristics, school performance, other clinical manifestations, and laboratory findings. RESULTS: Of the 145 patients with 22q11.2DS, 22 (15.2%) had epileptic seizures, 15 (10.3%) had developmental delay, and 5 (3.4%) had a psychiatric illness. Twelve patients with epilepsy were classified as genetic epilepsy whereas the remaining were classified as structural, including three with malformations of cortical development. Patients with epilepsy were more likely to display developmental delay (odds ratio=3.98; 95% confidence interval=1.5-10.5; p=0.005), and developmental delay was more common in patients with structural epilepsy than in those with genetic epilepsy. CONCLUSIONS: Patients with 22q11.2DS have a high risk of epilepsy, which in these cases is closely related to other NP manifestations. This implies that this specific genetic locus is critically linked to neurodevelopment and epileptogenesis.

Enzymatic and microbial transformation assays for the evaluation of the environmental fate of diclofenac and its metabolites.

Diclofenac has been of environmental concern due to the potential harmful effects on non-target organisms at environmentally relevant concentrations. In this study, we evaluated the transformation kinetics of diclofenac and its two major metabolites in two laboratory-scale experiments: the transformation of diclofenac in the presence of rat liver S9 fraction with co-factors, and the transformation of diclofenac, 4'-hydroxy-diclofenac and diclofenac ß-O-acyl glucuronide in the inoculum used for the OECD 301C ready-biodegradability test. 4'-Hydroxy-diclofenac was identified as the major phase I metabolite and diclofenac ß-O-acyl glucuronide was identified as the major phase II metabolite in the S9 assay. Transformation of diclofenac in the microbial degradation test did not occur significantly for 28 d, whereas 4'-hydroxy-diclofenac degraded slowly, indicating that the biological removal of diclofenac is not likely to occur in conventional STPs unless sorptive removal is significant. However, diclofenac ß-O-acyl glucuronide deconjugated to form equimolar diclofenac within 7 d, in the microbial degradation test. The mixture of diclofenac and its two metabolites, formed after incubating diclofenac in S9 medium for 2 h, was spiked in the inoculum to link both assays. The concentrations of diclofenac and its metabolites, measured over time, agreed well with predicted values, using rate parameters obtained from independent experiments. The results show that phase II metabolites generated in mammals may deconjugate easily in conventional STPs to form a parent compound and that these processes should be considered during the environmental monitoring and risk assessment of diclofenac.