Levels, spatial distributions, and provision of petroleum hydrocarbons and phthalates in sediments from Obhur lagoon, Red Sea coast of Saudi Arabia.

The levels, spatial distribution, and sources of petroleum hydrocarbons and phthalates were assessed in surface sediment samples from the urban lagoon of Obhur near Jeddah, the largest city on the Red Sea coast of Saudi Arabia. The lagoon was divided into the inner zone, middle zone, and outer zone based on its geomorphological features and developmental activities. n-Alkanes, hopane and sterane biomarkers, and unresolved complex mixture were the major petroleum hydrocarbon compounds of the total extractable organic matter. Phthalates were also measured in the sediment samples. In the three zones, n-alkanes ranged from 89.3 ± 88.5 to 103.2 ± 114.9 ng/g, whereas the hopane and sterane biomarkers varied from 69.4 ± 75.3 to 77.7 ± 69.9 ng/g and 72.5 ± 77.9-89.5 ± 82.2 ng/g, respectively. The UCM concentrations ranged from 821 ± 1119 to 1297 ± 1684 ng/g and phthalates from 37.4 ± 34.5 65 ± 68 ng/g. The primary origins of these anthropogenic hydrocarbons in the lagoon sediments were petroleum products (boat engine discharges, boat washing, lubricants, and wastewater flows) and plasticizers (plastic waste and litter). The proportions of anthropogenic hydrocarbons derived from petroleum products in the sediment's TEOM ranged from 43 ± 33 to 62 ± 15%, while the percentages for plasticizers varied from 2.9 ± 1.2 to 4.0 ± 1.6%. The presence and inputs of these contaminants from petroleum and plastic wastes in the lagoon's sediments will eventually have an impact on its habitats, including the benthic nursery and spawning areas.

Inflammation of Gill Epithelia in Fish Causes Increased Permeation of Petrogenic Polar Organic Chemicals via Disruption of Tight Junctions.

The epithelial cell layer that lines the gills of fish controls paracellular permeation of chemicals through tight junctions. The integrity of tight junctions can be affected by inflammation, which likely affects the bioavailability of chemicals. Here, the inflammation of the rainbow trout gill cell line RTgill-W1 was induced via exposure to bacterial lipopolysaccharides (LPS). Cells were then coexposed to extracts of oil sands process-affected water (OSPW), which contain complex mixtures of chemicals. After 24 h of exposure, cells exposed to LPS showed a reduction in transepithelial electrical resistance, an indicator of tight junction integrity. Quantitative reverse-transcription polymerase chain reaction (RT-PCR) analysis determined that abundances of transcripts of genes coding for tight junction proteins were significantly less in cells exposed to 20, 50, or 100 mg L-1 LPS. Chemical analysis revealed increased permeation of constituents of OSPW across epithelia at all studied LPS concentrations. These in vitro findings were confirmed in vivo in rainbow trout exposed to LPS and OSPW for 48 h, which resulted in greater accumulation of chemicals relative to that for fish exposed to OSPW alone. Our results demonstrated that inflammation and disruption of tight junctions could lead to greater uptake of potentially harmful chemicals from the environment, which has implications for mixture risk assessment.

Toxicokinetic Models for Bioconcentration of Organic Contaminants in Two Life Stages of White Sturgeon (Acipenser transmontanus).

The white sturgeon (Acipenser transmontanus) is an endangered ancient fish species that is known to be particularly sensitive to certain environmental contaminants, partly because of the uptake and subsequent toxicity of lipophilic pollutants prone to bioconcentration as a result of their high lipid content. To better understand the bioconcentration of organic contaminants in this species, toxicokinetic (TK) models were developed for the embryo-larval and subadult life stages. The embryo-larval model was designed as a one-compartment model and validated using whole-body measurements of benzo[a]pyrene (B[a]P) metabolites from a waterborne exposure to B[a]P. A physiologically based TK (PBTK) model was used for the subadult model. The predictive power of the subadult model was validated with an experimental data set of four chemicals. Results showed that the TK models could accurately predict the bioconcentration of organic contaminants for both life stages of white sturgeon within 1 order of magnitude of measured values. These models provide a tool to better understand the impact of environmental contaminants on the health and the survival of endangered white sturgeon populations.

Mechanisms of pH-Dependent Uptake of Ionizable Organic Chemicals by Fish from Oil Sands Process-Affected Water (OSPW).

Uptake and effects of ionizable organic chemicals (IOCs) that are weak acids in aqueous solution by fish can differ as a function of pH. While the pH-dependent behavior of select IOCs is well-understood, complex mixtures of IOCs, e.g., from oil sands process-affected water (OSPW), have not yet been studied systematically. Here, we established an in vitro screening method using the rainbow trout gill cell line, RTgill-W1, to investigate pH-dependent cytotoxicity and permeation of IOCs across cultured epithelia using ultra-high-performance liquid chromatography with high-resolution mass spectrometry (UPLC-HRMS). The assay was benchmarked using model chemicals and technical mixtures, and then used to characterize fractions and reconstituted extracts of field-collected OSPW. Significant pH-dependent cytotoxicity of individual IOCs, acidic fractions, and reconstituted extracts of OSPW was observed. In vitro data were in good agreement with data from a 96 h in vivo exposure experiment with juvenile rainbow trout. Permeation of some IOCs from OSPW was mediated by active transport, as revealed by studies in which inhibitors of these active transport mechanisms were applied. We conclude that the RTgill-W1 in vitro assay is useful for the screening of pH-dependent uptake of IOCs in fish, and has applications for in vitro-in vivo extrapolation, and prioritization of chemicals in nontarget screenings.

Toxicokinetics of Brominated Azo Dyes in the Early Life Stages of Zebrafish (Danio rerio) Is Prone to Aromatic Substituent Changes.

Brominated azo dyes (BADs) have been identified as predominant indoor brominated pollutants in daycare dust; thus, their potential health risk to children is of concern. However, the toxicities of BADs remain elusive. In this study, the toxicokinetics of two predominant BADs, Disperse Blue 373 (DB373) and Disperse Violet 93 (DV93), and their suspect metabolite 2-bromo-4,6-dinitroaniline (BDNA) was investigated in embryos of zebrafish (Danio rerio). The bioconcentration factor of DV93 at 120 hpf is 6.2-fold lower than that of DB373. The nontarget analysis revealed distinct metabolism routes between DB373 and DV93 by reducing nitro groups to nitroso (DB373) or amine (DV93), despite their similar structures. NAD(P)H quinone oxidoreductase 1 (NQO1) and pyruvate dehydrogenase were predicted as the enzymes responsible for the reduction of DB373 and DV93 by correlating time courses of the metabolites and enzyme development. Further in vitro recombinant enzyme and in vivo inhibition results validated NQO1 as the enzyme specifically reducing DB373, but not DV93. Global proteome profiling revealed that the expression levels of proteins from the "apoptosis-induced DNA fragmentation" pathway were significantly upregulated by all three BADs, supporting the bioactivation of BADs to mutagenic aromatic amines. This study discovered the bioactivation of BADs via distinct eukaryotic enzymes, implying their potential health risks.

Identification of Chemicals that Cause Oxidative Stress in Oil Sands Process-Affected Water.

Oil sands process-affected water (OSPW) has been reported to cause oxidative stress in organisms, yet the causative agents remain unknown. In this study, a high-throughput in vitro Nrf2 reporter system was used, to determine chemicals in OSPW that cause oxidative stress. Five fractions, with increasing polarity, of the dissolved organic phase of OSPW were generated by use of solid phase extraction cartridges. The greatest response of Nrf2 was elicited by F2 (2.7 ± 0.1-fold), consistent with greater hydroperoxidation of lipids in embryos of Japanese medaka (Oryzias latipes) exposed to F2. Classic naphthenic acids were mainly eluted in F1, and should not be causative chemicals. When F2 was fractionated into 60 subfractions by use of HPLC, significant activation of Nrf2 was observed in three grouped fractions: F2.8 (1.30 ± 0.01-fold), F2.16 (1.34 ± 0.05-fold), and F2.25 (1.28 ± 0.15-fold). 54 compounds were predicted to be potential chemicals causing Nrf2 response, predominated by SO3+ and O3+ species. By use of high-resolution MS2 spectra, these SO3+ and O3+ species were identified as hydroxylated aldehydes. This study demonstrated that polyoxygenated chemicals, rather than classic NAs, were the major chemicals responsible for oxidative stress in the aqueous phase of OSPW.

Toxicokinetics and toxicodynamics of chlorpyrifos is altered in embryos of Japanese medaka exposed to oil sands process-affected water: evidence for inhibition of P-glycoprotein.

Oil sands process-affected water (OSPW) is generated during extraction of bitumen in the surface mining oil sands industry in Alberta, Canada. Studies were performed in vitro by use of Caco-2 cells, and in vivo with larvae of Japanese medaka (Oryzias latipes) to determine if organic compounds from the aqueous phase of OSPW inhibit ATP binding cassette protein ABCB1 (permeability-glycoprotein, P-gp). Neutral and basic fractions of OSPW inhibited activity of P-gp in Caco-2 cells by 1.9- and 2.0-fold, respectively, while the acidic fraction had the least effect. The organophosphate pesticides chlorpyrifos (a substrate of P-gp) and malathion (not a substrate of P-gp), were used as model chemicals to investigate inhibition of P-gp in larvae. Co-exposure to chlorpyrifos and an extract of OSPW containing basic and neutral compounds reduced survival of larvae to 26.5% compared to survival of larvae exposed only to chlorpyrifos, which was 93.7%. However, co-exposure to malathion and the extract of OSPW did not cause acute lethality compared to exposure only to malathion. Accumulation and bioconcentration of chlorpyrifos, but not malathion, was greater in larvae co-exposed with the extract of OSPW. The terminal elimination half-life of chlorpyrifos in larvae exposed to chlorpyrifos in freshwater was 5 days compared with 11.3 days in larvae exposed to chlorpyrifos in OSPW. Results suggest that in non-acute exposures, basic and neutral organic compounds in the water-soluble fraction of OSPW inhibit activity of P-gp, which suggests that OSPW has the potential to cause adverse effects by chemosensitization. Copyright © 2016 John Wiley & Sons, Ltd.

Peroxisome Proliferator-Activated Receptor γ is a Sensitive Target for Oil Sands Process-Affected Water: Effects on Adipogenesis and Identification of Ligands.

Identification of toxic components of complex mixtures is a challenge. Here, oil sands process-affected water (OSPW) was used as a case study to identify those toxic components with a known protein target. Organic chemicals in OSPW exhibited dose-dependent activation of peroxisome proliferator-activated receptor γ (PPARγ) at concentrations less than those currently in the environment (0.025× equivalent of full-strength OSPW), by use of a luciferase reporter gene assay. Activation of PPARγ-mediated adipogenesis by OSPW was confirmed in 3T3L1 preadipocytes, as evidenced by accumulation of lipids and up-regulation of AP2, LPL, and PPARγ gene expression after exposure to polar fractions of OSPW. Unexpectedly, the nonpolar fractions of OSPW inhibited differentiation of preadipocytes via activation of the Wnt signaling pathway. Organic chemicals in OSPW that were ligands of PPARγ were identified by use of a pull-down system combined with untargeted chemical analysis (PUCA), with a recombinant PPARγ protein. Thirty ligands of PPARγ were identified by use of the PUCA assay. High resolution MS(1) and MS(2) spectra were combined to predict the formulas or structures of a subset of ligands, and polyoxygenated or heteroatomic chemicals, especially hydroxylated carboxylic/sulfonic acids, were the major ligands of PPARγ.

Densitometric validation and analysis of biomarker ß-amyrin in different Acacia species (leaves) grown in Kingdom of Saudi Arabia by high performance thin-layer chromatography.

Biomarker ß-amyrin was analyzed in the leaves of four different Acacia species (A. salicina, A. loreta, A. hamulosa and A. tortilis) grown in Kingdom of Saudi Arabia by a validated HPTLC method. The chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates using solvents toluene: methanol (9:1, v/v) as mobile phase. The developed TLC plate was derivatized with anisaldehyde and scanned at 520 nm. A sharp peak of ß-amyrin was found at Rf=0.58±0.01. The r2 and the linear regression equation for ß-amyrin was found to be 0.991 and 19.913X+107.803, respectively in the concentration range of 100-800 ng. The percentage of ß-amyrin was found to be maximum 2.70% w/w in A. tortilis, 1.85% w/w in A. loreta and 1.80% w/w in A. hamulosa while it was totally absent in A. salicina. This study conceives maiden reporting of quantification of ß-amyrin in four different species of Acacia by validated HPTLC method. The developed method for the analysis of ß-amyrin was proved to be reproducible by statistical analysis hence it can be employed for further analysis of ß-amyrin in plasma, other biological fluids and in finished products available in the market.

Silencing sensory neuron membrane protein RferSNMPu1 impairs pheromone detection in the invasive Asian Palm Weevil.

The red palm weevil (RPW), Rhynchophorus ferrugineus (Olivier), also known as the Asian palm weevil, is an invasive pest that causes widespread damage to palm trees around the globe. As pheromone communication is crucial for their mass attack and survival on palm trees, the olfactory concept of pest control strategies has been widely explored recently. We aim to understand the molecular basis of olfaction in RPW by studying one of the key olfactory proteins in insect pheromone communication, sensory neuron membrane proteins (SNMPs). SNMPs belong to the CD36 (cluster of differentiation 36) family that perform two distinct olfactory roles in insects, either in pheromone (odorant) transfer to the odorant receptors (SNMP1) or in the pheromone clearing process (SNMP2). In this study, we performed antennal transcriptomic screening and identified six SNMPs, mapping them on the R. ferrugineus genome, and confirmed four distinct SNMPs. Both SNMP1 proteins in RPW, viz., RferSNMPu1 and RferSNMPu2, were mapped onto the same scaffold in different loci in the RPW genome. To further understand the function of these proteins, we first classified them using phylogenetic analysis and checked their tissue-specific expression patterns. Further, we measured the relative transcript abundance of SNMPs in laboratory-reared, field-collected adults and pheromone-exposure experiments, ultimately identifying RferSNMPu1 as a potential candidate for functional analysis. We mapped RferSNMPu1 expression in the antennae and found that expression patterns were similar in both sexes. We used RNAi-based gene silencing to knockdown RferSNMPu1 and tested the changes in the RPW responses to aggregation pheromone compounds, 4-methyl-5-nonanol (ferrugineol) and 4-methyl-5-nonanone (ferrugineone), and a kairomone, ethyl acetate using electroantennogram (EAG) recordings. We found a significant reduction in the EAG recordings in the RferSNMPu1 knockdown strain of adult RPWs, confirming its potential role in pheromone detection. The structural modelling revealed the key domains in the RferSNMPu1 structure, which could likely be involved in pheromone detection based on the identified ectodomain tunnels. Our studies on RferSNMPu1 with a putative role in pheromone detection provide valuable insight into understanding the olfaction in R. ferrugineus as well as in other Curculionids, as SNMPs are under-explored in terms of its functional role in insect olfaction. Most importantly, RferSNMPu1 can be used as a potential target for the olfactory communication disruption in the R. ferrugineus control strategies.

Polycyclic Aromatic Hydrocarbons (PAHs) and Metals in Diverse Biochar Products: Effect of Feedstock Type and Pyrolysis Temperature.

Biochar's agricultural and environmental benefits have been widely demonstrated; however, it may cause environmental contamination if it contains large amounts of pollutants such as polycyclic aromatic hydrocarbons (PAHs) and heavy metals (HMs). Therefore, this study aimed to assess the contents of PAHs and HM in a range of biochars generated from different sources and pyrolysis temperatures. A range of feedstock was converted to biochar, including sewage sludge (SS), olive mill pomace (OP), feather meal (FM), soft offal meal (CSM), chicken manure (CM), and date palm residues (DPR). Each feedstock was then pyrolyzed at three temperatures of 300, 500, or 700 °C, thereby producing a total of 18 types of biochar. These biochar products were analyzed for 16 PAHs and eight metals (Cr, Mn, Fe, Ni, Cu, Zn, Cd, and Pb). Benzo[b]fluoranthene, benzo[k]fluoranthene, and benzo(a)pyrene were significantly greater in the biochar produced at 700 °C than in that produced at 300 °C, especially for CM. The concentrations of dibenz(a,h)anthracene were significantly lower at 700 °C but greater at 500 °C and 300 °C in DPR. Increasing the pyrolysis temperature from 300 to 700 °C significantly increased the concentrations of metals, including Cr in SS and OP; Mn in CM; and Fe, Ni, Cu, and Zn in SS. However, the concentration of Cd was significantly lower in the SS when biochar was produced at 700 °C than at 500 or 300 °C. The type of feedstock used and the pyrolysis temperature are key factors influencing the contents of PAHs and HMs in biochar, both of which need to be considered during the production and use of biochar. Further investigations are recommended to establish the relationships between pyrolysis temperature and types of feedstock and the formation of PAH or the concentrations of metals. Monitoring the concentrations of PAHs and HMs before applying biochar to soil is also recommended.

Correction to "Mesoporous Activated Carbon from Leaf Sheath Date Palm Fibers by Microwave-Assisted Phosphoric Acid Activation for Efficient Dye Adsorption".

[This corrects the article DOI: 10.1021/acsomega.2c03755.].

Production of activated carbon from date palm stones by hydrothermal carbonization and microwave assisted KOH/NaOH mixture activation for dye adsorption.

Date palm stones are regarded as possible alternatives to activated carbon (AC) precursors with high potential for various environmental applications. In this research study, date palm stones derived activated carbon (DPSAC) was used as adsorbent for removing toxic remazol brilliant blue R (RBBR). The synthesis of DPSAC involved a chemical treatment using KOH and NaOH (1:1). Characterization of DPSAC revealed that it exhibited a BET surface area of 715.30 m2/g, Langmuir surface area of 1061.93 m2/g, total pore volume of 0.39 cm3/g, and average pore diameter of 2.15 nm. Adsorption uptake of RBBR increased (from 24.54 to 248.54 mg/g), whereas the removal percentage decreased (from 98.16 to 82.85%) when the initial RBBR concentration increased (from 25 to 300 mg/L). The adsorption process performed best under acidic conditions (pH 3), with an RBBR uptake of 98.33 mg/g. Because of the high R2 values (0.9906 and 0.9779) and low average errors (6.24 and 13.95%), this adsorption process followed the Freundlich isotherm and pseudo-first-order (PFO) models, respectively. The Langmuir adsorption capacity (Qm) was 319.63 mg/g. Thermodynamic parameters were - 11.34 kJ/mol for ∆H° (exothermic in nature), 0.05 kJ/mol K for ∆S° (increasing randomness level at solid-liquid interface), - 27.37 kJ/mol for ∆G° (spontaneous), and 6.84 kJ/mol for Ea (controlled by physisorption).

Functional Studies and Revision of the NFAT-133/TM-123 Biosynthetic Pathway in Streptomyces pactum.

The biosynthetic gene cluster of NFAT-133, an inhibitor of the nuclear factor of activated T cells, was recently identified in Streptomyces pactum ATCC 27456. This cluster is conspicuous by its highly disordered noncollinear type I modular polyketide synthase (PKS) genes that encode PKSs with one module more than those expected for the heptaketide NFAT-133 biosynthesis. Thus, the major metabolite NFAT-133 was proposed to derive from an octaketide analogue, TM-123. Here, we report that further bioinformatic analysis and gene inactivation studies suggest that NFAT-133 is not derived from TM-123 but rather a product of programmed KS7 extension skipping of a nascent heptaketide from the PKS assembly line that produces TM-123. Furthermore, identification of NFAT-133/TM-123 analogues from mutants of the ATCC 27456 strain suggests that NftN (a putative dehydrogenase), NftE (a cytochrome P450), and NftG (a putative hydrolase/decarboxylase) function "in trans" during the polyketide chain assembly processes.

Mesoporous Activated Carbon from Leaf Sheath Date Palm Fibers by Microwave-Assisted Phosphoric Acid Activation for Efficient Dye Adsorption.

Remazol Brilliant Blue R (RBBR) is a common dye used in the industry, and its presence in wastewater and discharge into the environment can create a serious concern for the ecosystem and human health. Activated carbon produced from crop residues has emerged as a promising technique for removing contaminants from wastewater. In this study, leaf sheath date palm fiber-based activated carbon (LSDAC) was synthesized via phosphoric acid, H3PO4, treatment, followed by a microwave-induced carbonization process. The produced LSDAC was found to have a BET surface area of 604.61 m2/g, a Langmuir surface area of 922.05 m2/g, a total pore volume of 0.35 cm3/g, and an average pore size of 2.75 nm. The highest removal of RBBR was achieved at a solution pH of 3 (92.56 mg/g) and a solution temperature of 50 °C (90.37 mg/g). Adsorption of RBBR onto LSDAC followed the Langmuir isotherm model with a maximum monolayer capacity, Q m, of 243.43 mg/g, whereas in terms of kinetics, this adsorption system was best described by the pseudo-first-order (PFO) model. The calculated thermodynamic parameters ΔH°, ΔS°, ΔG°, and Arrhenius activation energy, E a, were 4.71 kJ/mol, 0.10 kJ/mol·K, -26.25 kJ/mol, and 5.88 kJ/mol, respectively, indicating that the adsorption of RBBR onto LSDAC was endothermic in nature, exhibited increased randomness at the solid-liquid interface, and was spontaneous and controlled by physisorption.

Catalytic efficiency of soil enzymes explains temperature sensitivity: Insights from physiological theory.

Soil enzymes are crucial for carbon and nutrient cycling and are highly sensitive to warming. Biochemical reaction rates increase with temperature according to the Arrhenius law, but changes in microbial physiology may partially counteract this warming-induced acceleration that leads enzymatic rates to deviate from Arrhenius law. Here, we attempt to reconcile disparate views on the enzyme responses to warming based on the Arrhenius law and physiological theory by enzyme catalytic efficiency. In this study, we tested the kinetic parameters of five key enzymes of C, N, and P cycling to warming (from 0 to 40 °C) in cropland soils originating from 5 different temperate zones. The soils were incubated for one month at 0, 10, 20, 30, and 40 °C (±0.5 °C) with 60% water holding capacity (WHC). The kinetic parameters were calculated and measured at a range of 4-methyumbelliferone (MUB)-substrate concentrations. We found that catalytic efficiency (Vmax/Km) of individual enzymes ranged from 0.05 to 27 s-1 between 0 and 40 °C. Maximum reaction rate (Vmax) increased with warming, while Vmax/Km of most enzymes remained stable by warming at low temperatures (up to 10 °C), and it raised from 20 to 40 °C. Most enzymes had lower substrate affinities (Km) and increased their efficiency with warming. Consistent with studies considering Arrhenius law solely, the temperature sensitivity (Q10) of Vmax decreased with warming. However, the Q10 of Vmax/Km displayed a lower value in the cold but a higher value in warmer temperature, which confirmed microbial adaptation based on physiological theory, consequently encouraging its linking with the Arrhenius law. Therefore, Arrhenius linked with physiological theory could correct explanation of enzyme activities by warming. Considering the microbial adaptation to temperature, the present predicted warming-induced acceleration of soil organic matter decomposition might be overestimated in cold and underestimated in warm environments.

Modulation of Specialized Metabolite Production in Genetically Engineered Streptomyces pactum.

Filamentous soil bacteria are known to produce diverse specialized metabolites. Despite having enormous potential as a source of pharmaceuticals, they often produce bioactive metabolites at low titers. Here, we show that inactivation of the pactamycin, NFAT-133, and conglobatin biosynthetic pathways in Streptomyces pactum ATCC 27456 significantly increases the production of the mitochondrial electron transport inhibitors piericidins. Similarly, inactivation of the pactamycin, NFAT-133, and piericidin pathways significantly increases the production of the heat-shock protein (Hsp) 90 inhibitor conglobatin. In addition, four new conglobatin analogues (B2, B3, F1, and F2) with altered polyketide backbones, together with the known analogue conglobatin B1, were identified in this mutant, indicating that the conglobatin biosynthetic machinery is promiscuous toward different substrates. Among the new conglobatin analogues, conglobatin F2 showed enhanced antitumor activity against HeLa and NCI-H460 cancer cell lines compared to conglobatin. Conglobatin F2 also inhibits colony formation of HeLa cells in a dose-dependent manner. Molecular modeling studies suggest that the new conglobatins bind to human Hsp90 and disrupt Hsp90/Cdc37 chaperone/co-chaperone interactions in the same manner as conglobatin. The study also showed that genes that are involved in piericidin biosynthesis are clustered in two different loci located distantly in the S. pactum genome.

Microbial Communities and Functions in the Rhizosphere of Disease-Resistant and Susceptible Camellia spp.

Oil tea (Camellia spp.) is endemic to the hilly regions in the subtropics. Camellia yuhsienensis is resistant to diseases such as anthracnose and root rot, while Camellia oleifera is a high-yield species but susceptible to these diseases. We hypothesize that differences in the rhizosphere microbial communities and functions will elucidate the resistance mechanisms of these species. We used high-throughput sequencing over four seasons to characterize the rhizosphere microbiome of C. oleifera (Rhizo-Sus) and C. yuhsienensis (Rhizo-Res) and of the bulk soil control (BulkS). In Rhizo-Res, bacterial richness and diversity (Shannon index) in autumn and winter were both higher than that in Rhizo-Sus. In Rhizo-Res, fungal richness in autumn and winter and diversity in summer, autumn, and winter were higher than that in Rhizo-Sus. The seasonal variations in bacterial community structure were different, while that of fungal community structure were similar between Rhizo-Res and Rhizo-Sus. Gram-positive, facultatively anaerobic, and stress-tolerant bacteria were the dominant groups in Rhizo-Sus, while Gram-negative bacteria were the dominant group in Rhizo-Res. The significant differences in bacterial and fungal functions between Rhizo-Sus and Rhizo-Res were as follows: (1) in Rhizo-Sus, there were three bacterial and four fungal groups with plant growth promoting potentials, such as Brevibacterium epidermidis and Oidiodendron maius, and one bacterium and three fungi with pathogenic potentials, such as Gryllotalpicola sp. and Cyphellophora sessilis; (2) in Rhizo-Res, there were also three bacteria and four fungal groups with plant-growth-promoting potentials (e.g., Acinetobacter lwoffii and Cenococcum geophilum) but only one phytopathogen (Schizophyllum commune). In summary, the rhizosphere microbiome of disease-resistant C. yuhsienensis is characterized by a higher richness and diversity of microbial communities, more symbiotic fungal communities, and fewer pathogens compared to the rhizosphere of high-yield but disease-susceptible C. oleifera.

In vitro-in vivo and cross-life stage extrapolation of uptake and biotransformation of benzo[a]pyrene in the fathead minnow (Pimephales promelas).

Understanding internal dose metrics is integral to adequately assess effects environmental contaminants might have on aquatic wildlife, including fish. In silico toxicokinetic (TK) models are a leading approach for quantifying internal exposure metrics for fishes; however, they often do not adequately consider chemicals that are actively biotransformed and have not been validated against early-life stages (ELS) that are often considered the most sensitive to the exposure to contaminants. To address these uncertainties, TK models were parameterized for the rapidly biotransformed chemical benzo[a]pyrene (B[a]P) in embryo-larval and adult life stages of fathead minnows. Biotransformation of B[a]P was determined through measurements of in vitro clearance. Using in vitro-in vivo extrapolation, in vitro clearance was integrated into a multi-compartment TK model for adult fish and a one-compartment model for ELS. Model predictions were validated using measurements of B[a]P metabolites from in vivo flow-through exposures to graded concentrations of water-borne B[a]P. Significantly greater amounts of B[a]P metabolites were observed with exposure to greater concentrations of parent compound in both life stages. However, when assessing biotransformation capacity, no differences in phase I or phase II biotransformation were observed with greater exposures to B[a]P. Results of modelling suggested that biotransformation of B[a]P can be successfully implemented into in silico models to accurately predict life stage-specific abundances of B[a]P metabolites in either whole-body larvae or the bile of adult fish. Models developed increase the scope of applications in which TK models can be used to support environmental risk assessments.

Organophosphate pesticide method development and presence of chlorpyrifos in the feet of nearctic-neotropical migratory songbirds from Canada that over-winter in Central America agricultural areas.

Recent modeling analysis suggests that numerous birds may be at risk of acute poisoning in insecticide-treated fields. Although the majority of avian field studies on pesticides have focused on treated seed, granule, insect or vegetation (oral exposure) ingestion, dermal exposure is an important exposure route when birds come into contact with deposited pesticides on foliage and other surfaces. Some nearctic-neotropical migratory songbirds are likely exposed to pesticides on their non-breeding habitats and include treated crops, plantations or farmlands. In the present study, we developed a method for four environmentally-relevant organophosphate (OP) pesticides (fenthion, fenamiphos, chlorpyrifos and diazinon) in the feet of migratory songbirds (i.e. Common yellowthroat, Gray catbird, Indigo bunting, America redstart, Northern waterthrush, Northern parula, and an additional 12 species of warblers). A total of 190 specimens of the 18 species of songbirds were sampled from available window-killed birds (spring of 2007 and 2011) in downtown Toronto, Canada. The species that were available most likely over-wintered in Mexican/Central American crops such as citrus, coffee and cacao. The feet of the dead birds were sampled and where OP foot exposure likely occurred during over-wintering foraging on pesticide-treated crops. Chlorpyrifos was the only measurable OP (pg mg feet weight(-1)) and in the 2011-collected feet of Black throated blue warbler (0.5), Tennessee warbler (1.0), Northern parula (1.2), Northern waterthrush (0.6), Common yellowthroat (1.0) and the Blue winged warbler (0.9). Dermal contact with OP pesticides during over-wintering in agricultural areas resulted in low levels of chlorpyrifos and long time retention on the feet of a subset of songbirds.