Association between N, N-diethyl-m-toluamide exposure and the odds of kidney stones in US adults: a population-based study.

Background: Currently, there is limited research on the specific relationship between N, N-diethyl-m-toluamide (DEET) exposure and the odds of kidney stones. We aimed to investigate the relationship between DEET exposure and the prevalence of kidney stones. Methods: We included 7,567 qualified participants in our research from the 2007-2016 NHANES survey. We carried out three logistic regression models to explore the potential association between DEET exposure and the odds of kidney stones. Spline smoothing with generalized additive models (GAM) was utilized to assess the non-linear relationship and restricted cubic spline (RCS) curves was to determine the dose-response association. Multivariate regression models were used to conduct stratified analysis and sensitivity analysis. Results: Baseline characteristics of study participants presented the distribution of covariables. Regression analysis revealed that the odds of kidney stones were positively associated with the main metabolites of 3-diethyl-carbamoyl benzoic acid (DCBA) (log2) (OR = 1.05, 95% CI 1.02 to 1.08). The fourth quartile of urine DCBA showed a greater risk of kidney stones in the fully adjusted model (OR = 1.36, 95% CI 1.08 to 1.72). Another DEET metabolite of N, N-diethyl-3-hydroxymethylbenzamide (DHMB) was used to confirm the accuracy and stability of the results. The spline smoothing curve represented two main DEET metabolites had similar no-linear relationships and a positive trend with kidney stones proportion. RCS implied that the incidence of kidney stones rose with increasing levels of DEET exposure. High-risk groups on kidney stones were exhibited by stratified analysis under DEET exposure. Conclusion: Our study suggests that DEET exposure is positively associated with odds of kidney stones. Further investigation into the underlying processes of this association is required to guide the prevention and treatment of kidney stones.

The structure of AgamOBP5 in complex with the natural insect repellents Carvacrol and Thymol: Crystallographic, fluorescence and thermodynamic binding studies.

Among several proteins participating in the olfactory perception process of insects, Odorant Binding Proteins (OBPs) are today considered valid targets for the discovery of compounds that interfere with their host-detection behavior. The 3D structures of Anopheles gambiae mosquito AgamOBP1 in complex with the known synthetic repellents DEET and Icaridin have provided valuable information on the structural characteristics that govern their selective binding. However, no structure of a plant-derived repellent bound to an OBP has been available until now. Herein, we present the novel three-dimensional crystal structures of AgamOBP5 in complex with two natural phenolic monoterpenoid repellents, Carvacrol and Thymol, and the MPD molecule. Structural analysis revealed that both monoterpenoids occupy a binding site (Site-1) by adopting two alternative conformations. An additional Carvacrol was also bound to a secondary site (Site-2) near the central cavity entrance. A protein-ligand hydrogen-bond network supplemented by van der Waals interactions spans the entire binding cavity, bridging α4, α6, and α3 helices and stabilizing the overall structure. Fluorescence competition and Differential Scanning Calorimetry experiments verified the presence of two binding sites and the stabilization effect on AgamOBP5. While Carvacrol and Thymol bind to Site-1 with equal affinity in the submicromolar range, they exhibit a significantly lower and distinct binding capacity for Site-2 with Kd's of ~7 µΜ and ~18 µΜ, respectively. Finally, a comparison of AgamOBP5 complexes with the AgamOBP4-Indole structure revealed that variations of ligand-interacting aminoacids such as A109T, I72M, A112L, and A105T cause two structurally similar and homologous proteins to display different binding specificities.

Comprehensive Molecular Interaction Studies to Construe the Repellent/Kill Activity of Geraniol During Binding Event Against Aedes aegypti Proteins.

Aedes aegypti is an etiological agent for dengue, chikungunya, zika, and yellow fever viruses. With the advent of the use of natural alternatives as repellents, their precise mode of action during the event of binding is still unclear. Geraniol is one such bioactive natural that has been previously shown to have some insecticide properties. Thus, the present study aimed to understand the mechanism of the binding event of geraniol with the whole proteome of A. aegypti. Twenty protein target categories were shortlisted for the mosquito, wherein the proteins were downloaded with respect to the reference proteome. Conserved domain analysis was performed for the same using the CDD search tool to find the proteins that have common domains. 309 proteins were modeled using RaptorX standalone tool, and validated using Ramachandran plots from SAVES v6.0 from ProCheck. These modeled and validated proteins were then docked against geraniol, using POAP software, for understanding the binding energies. The top 3 best-docked complexes were then analyzed for their stabilities and event of binding via 100 ns simulation studies using DESMOND's Maestro environment. The docking results showed that the geraniol-voltage-gated sodium channel had the best energy of - 7.1 kcal/mol, followed by geraniol-glutathione-S-transferase (- 6.8 kcal/mol) and geraniol-alpha esterase (- 6.8 kcal/mol). The simulations for these 3 complexes revealed that several residues of the proteins interacted well with geraniol at a molecular level, and all three docked complexes were found to be stable when simulated (RMSD: 16-18 Å, 3.6-4.8 Å, 4.8-5.6 Å, respectively). Thus, the present study provides insights into the mechanism of the binding event of geraniol with the major A. aegypti targets, thereby, assisting the use of geraniol as a natural repellent.

Microencapsulation of DEET in Solid Lipid Microparticles: production, characterization and safety evaluation.

DEET is considered the gold standard for insect repellent products. However, it behaves as a strong skin permeant. DEET was encapsulated in Solid Lipid Microparticles (SLM) and characterized in terms of morphology, particle size, cytotoxicity and ex vivo permeation. The particles exhibited micrometric size with a spherical shape. In addition, we developed and validated an analytical method for DEET quantification by high performance liquid chromatography (HPLC), which was selective, linear, precise, accurate and robust. The toxicity test in cell culture of keratinocytes, fibroblasts and macrophages showed that the formulation did not present cytotoxicity. The SLM were able to decrease the skin permeation of DEET in relation to the free active in ethanol with gain in the safe. Microparticles were able to increase the skin retention of DEET, which can contribute to extend the time of repellent action. The results showed that Solid Lipid Microparticles are safe and promising topical formulation to insect bite prevention.

Engineering Yeast for De Novo Synthesis of the Insect Repellent Nepetalactone.

While nepetalactone, the active ingredient in catnip, is a potent insect repellent, its low in planta accumulation limits its commercial viability as an alternative repellent. Here we describe for the first time de novo nepetalactone synthesis in Saccharomyces cerevisiae, enabling sustainable and scalable production. Nepetalactone production required introducing eight exogenous genes including the cytochrome P450 geraniol-8-hydroxylase, the bottleneck of the heterologous pathway. Combinatorial assessment of geraniol-8-hydroxylase and cytochrome P450 reductase variants, and copy-number variations were used to overcome this bottleneck. We found that several reductases improved hydroxylation activity and increasing geraniol-8-hydroxylase gene copy number improved 8-hydroxygeraniol titers. The accumulation of an unwanted metabolite implied inefficient channeling of carbon through the pathway. With the native yeast old yellow enzymes previously shown to use monoterpene intermediates as substrates, both homologues were deleted. These deletions increased 8-hydroxygeraniol yield, resulting in 3.10 mg/L/OD600 of nepetalactone from simple sugar in microtiter plates. This optimized pathway will benefit the development of high yielding strains for the scale up production of nepetalactone.

Volatiles Emission by Crotalaria nitens after Insect Attack.

Plants are known to increase the emission of volatile organic compounds upon the damage of phytophagous insects. However, very little is known about the composition and temporal dynamics of volatiles released by wild plants of the genus Crotalaria (Fabaceae) attacked with the specialist lepidopteran caterpillar Utetheisa ornatrix (Linnaeus) (Erebidae). In this work, the herbivore-induced plant volatiles (HIPV) emitted by Crotalaria nitens Kunth plants were isolated with solid phase micro-extraction and the conventional purge and trap technique, and their identification was carried out by GC/MS. The poly-dimethylsiloxane/divinylbenzene fiber showed higher affinity for the extraction of apolar compounds (e.g., trans-ß-caryophyllene) compared to the Porapak™-Q adsorbent from the purge & trap method that extracted more polar compounds (e.g., trans-nerolidol and indole). The compounds emitted by C. nitens were mainly green leaf volatile substances, terpenoids, aromatics, and aldoximes (isobutyraldoxime and 2-methylbutyraldoxime), whose maximum emission was six hours after the attack. The attack by caterpillars significantly increased the volatile compounds emission in the C. nitens leaves compared to those subjected to mechanical damage. This result indicated that the U. ornatrix caterpillar is responsible for generating a specific response in C. nitens plants. It was demonstrated that HIPVs repelled conspecific moths from attacked plants and favored oviposition in those without damage. The results showed the importance of volatiles in plant-insect interactions, as well as the choice of appropriate extraction and analytical methods for their study.

Identification of multiple odorant receptors essential for pyrethrum repellency in Drosophila melanogaster.

Pyrethrum extract from dry flowers of Tanacetum cinerariifolium (formally Chrysanthemum cinerariifolium) has been used globally as a popular insect repellent against arthropod pests for thousands of years. However, the mechanistic basis of pyrethrum repellency remains unknown. In this study, we found that pyrethrum spatially repels and activates olfactory responses in Drosophila melanogaster, a genetically tractable model insect, and the closely-related D. suzukii which is a serious invasive fruit crop pest. The discovery of spatial pyrethrum repellency and olfactory response to pyrethrum in D. melanogaster facilitated our identification of four odorant receptors, Or7a, Or42b, Or59b and Or98a that are responsive to pyrethrum. Further analysis showed that the first three Ors are activated by pyrethrins, the major insecticidal components in pyrethrum, whereas Or98a is activated by (E)-ß-farnesene (EBF), a sesquiterpene and a minor component in pyrethrum. Importantly, knockout of Or7a, Or59b or Or98a individually abolished fly avoidance to pyrethrum, while knockout of Or42b had no effect, demonstrating that simultaneous activation of Or7a, Or59b and Or98a is required for pyrethrum repellency in D. melanogaster. Our study provides insights into the molecular basis of repellency of one of the most ancient and globally used insect repellents. Identification of pyrethrum-responsive Ors opens the door to develop new synthetic insect repellent mixtures that are highly effective and broad-spectrum.

Characterization of a composite with enhanced attraction to savannah tsetse flies from constituents or analogues of tsetse refractory waterbuck (Kobus defassa) body odor.

Savannah tsetse flies avoid flying toward tsetse fly-refractory waterbuck (Kobus defassa) mediated by a repellent blend of volatile compounds in their body odor comprised of δ-octalactone, geranyl acetone, phenols (guaiacol and carvacrol), and homologues of carboxylic acids (C5-C10) and 2-alkanones (C8-C13). However, although the blends of carboxylic acids and that of 2-alkanones contributed incrementally to the repellency of the waterbuck odor to savannah tsetse flies, some waterbuck constituents (particularly, nonanoic acid and 2-nonanone) showed significant attractive properties. In another study, increasing the ring size of δ-octalactone from six to seven membered ring changed the activity of the resulting molecule (ε-nonalactone) on the savannah tsetse flies from repellency to attraction. In the present study, we first compared the effect of blending ε-nonalactone, nonanoic acid and 2-nonanone in 1:1 binary and 1:1:1 ternary combination on responses of Glossina pallidipes and Glossina morsitans morsitans tsetse flies in a two-choice wind tunnel. The compounds showed clear synergistic effects in the blends, with the ternary blend demonstrating higher attraction than the binary blends and individual compounds. Our follow up laboratory comparisons of tsetse fly responses to ternary combinations with different relative proportions of the three components showed that the blend in 1:3:2 proportion was most attractive relative to fermented cow urine (FCU) to both tsetse species. In our field experiments at Shimba Hills game reserve in Kenya, where G. pallidipes are dominant, the pattern of tsetse catches we obtained with different proportions of the three compounds were similar to those we observed in the laboratory. Interestingly, the three-component blend in 1:3:2 proportion when released at optimized rate of 13.71mg/h was 235% more attractive to G. pallidipes than a combination of POCA (3-n-Propylphenol, 1-Octen-3-ol, 4-Cresol, and Acetone) and fermented cattle urine (FCU). This constitutes a novel finding with potential for downstream deployment in bait technologies for more effective control of G. pallidipes, G. m. morsitans, and perhaps other savannah tsetse fly species, in 'pull' and 'pull-push' tactics.

Virus-induced plant volatiles mediate the olfactory behaviour of its insect vectors.

Plant viruses can manipulate their hosts to release odours that are attractive or repellent to their insect vectors. However, the volatile organic compounds (VOCs), either individually or as mixtures, which play a key role in the olfactory behaviour of insect vectors remains largely unknown. Our study focused on green rice leafhoppers (GRLHs) vectoring rice dwarf virus (RDV) revealed that RDV infection significantly induced the emission of (E)-ß-caryophyllene and 2-heptanol by rice plants, which influenced the olfactory behaviour of both non-viruliferous and viruliferous GRLHs. (E)-ß-caryophyllene attracted non-viruliferous GRLHs to settle on RDV-infected plants, but neither attracted nor repelled viruliferous GRLHs. In contrast, 2-heptanol repelled viruliferous GRLHs to settle on RDV-infected plants, but neither repelled nor attracted non-viruliferous GRLHs. Suppression of (E)-ß-caryophyllene synthase OsCAS via CRISPR-Cas9 to generate oscas-1 plants enabled us to confirm the important role played by (E)-ß-caryophyllene in modulating the virus-vector-host plant interaction. These novel results reveal the role of these virus-induced VOCs in modulating the behaviour of its GRLH insect vector and may facilitate the design of new strategies for disease control through manipulation of plant volatile emissions.

Antagonist effects of the leek Allium porrum as a companion plant on aphid host plant colonization.

Combining a non-host plant (companion plant or CP) with a target cultivated plant is considered as a promising strategy to reduce pest pressure. Among the companion plants (CP) commonly used in integrated systems, those belonging to the Amaryllidaceae family (chives, garlic, onion, leek) exhibit characteristics related to certain volatile organic compounds (VOCs) with promising repellent potentialities. The aim of this work was to investigate the potential disruption of sweet pepper (host plant) colonization by the green peach aphid (Myzus persicae) when exposed to leek (Allium porrum) as a CP. Retention/dispersion, EPG and clip-cage/Petri dish laboratory experiments were thus performed to study the effect of leek VOCs on aphid settlement/migration, feeding behavior and life history traits parameters, respectively. This work revealed that leek as a CP had a negative effect on aphid feeding behavior, by disturbing the balance between phloem and xylem sap ingestion, but had no influence concerning aphid settlement. Surprisingly, leek as a CP triggered some unexpected probiotic effects on certain life history traits such as aphid survival, biomass, and fecundity, suggesting a possible hormetic effect of leek VOCs on aphid physiology. The possibility of experience-induced preference of aphids for leek VOCs was also discussed.

Less Is More: a Mutation in the Chemical Defense Pathway of Erysimum cheiranthoides (Brassicaceae) Reduces Total Cardenolide Abundance but Increases Resistance to Insect Herbivores.

Erysimum cheiranthoides L (Brassicaceae; wormseed wallflower) accumulates not only glucosinolates, which are characteristic of the Brassicaceae, but also abundant and diverse cardenolides. These steroid toxins, primarily glycosylated forms of digitoxigenin, cannogenol, and strophanthidin, inhibit the function of essential Na+/K+-ATPases in animal cells. We screened a population of 659 ethylmethanesulfonate-mutagenized E. cheiranthoides plants to identify isolates with altered cardenolide profiles. One mutant line exhibited 66% lower cardenolide content, resulting from greatly decreased cannogenol and strophanthidin glycosides, partially compensated for by increases in digitoxigenin glycosides. This phenotype was likely caused by a single-locus recessive mutation, as evidenced by a wildtype phenotype of F1 plants from a backcross, a 3:1 wildtype:mutant segregation in the F2 generation, and genetic mapping of the altered cardenolide phenotype to one position in the genome. The mutation created a more even cardenolide distribution, decreased the average cardenolide polarity, but did not impact most glucosinolates. Growth of generalist herbivores from two feeding guilds, Myzus persicae Sulzer (Hemiptera: Aphididae; green peach aphid) and Trichoplusia ni Hübner (Lepidoptera: Noctuidae; cabbage looper), was decreased on the mutant line compared to wildtype. Both herbivores accumulated cardenolides in proportion to the plant content, with T. ni accumulating higher total concentrations than M. persicae. Helveticoside, a relatively abundant cardenolide in E. cheiranthoides, was not detected in M. persicae feeding on these plants. Our results support the hypothesis that increased digitoxigenin glycosides provide improved protection against M. persicae and T. ni, despite an overall decrease in cardenolide content of the mutant line.

Volatile Organic Compounds as Insect Repellents and Plant Elicitors: an Integrated Pest Management (IPM) Strategy for Glasshouse Whitefly (Trialeurodes vaporariorum).

The glasshouse whitefly (Trialeurodes vaporariorum Westwood) is a polyphagous arthropod pest that is of particular detriment to glasshouse grown tomato (Solanum lycopersicum) across temperate regions of the world. Control of whiteflies with synthetic pesticides has resulted in the evolution of resistant genotypes and a reduction in natural enemies, thus highlighting the need for environmentally sound control strategies. Volatile organic compounds (VOCs) offer an environmentally benign alternative to synthetic chemical sprays and this study explored the use of VOCs as insect repellents and plant defence elicitors to control whiteflies on tomato in a commercial glasshouse setting. Limonene in the form of a volatile dispenser system was found to successfully repel whitefly from the target crop and increased fruit yield by 32% during a heavy whitefly infestation. Analysis of tomato herbivore induced plant volatiles (HIPVs) led us to select methyl salicylate (MeSA) as the plant elicitor and application of MeSA to un-infested tomato plants was found to successfully reduce whitefly population development and increase yield by 11%, although this difference was marginally statistically significant. Combination of these two methods was also effective but whitefly abundance in combined plots was similar to the standalone limonene treatment across the course of the experiment. All of the VOC based control methods we used had a negative impact on whitefly performance, with more pronounced effects during the first few weeks of infestation. In subsequent laboratory experiments, we found elevated peroxidase (POD) activity and a significant increase in TPX1 and PR1 transcripts in MeSA treated plants. This led us to deduce that MeSA immediately induced plant defences, rather than priming them. We did however see evidence for residual priming, as plants treated with MeSA and infested with whiteflies produced significantly higher levels of POD activity than whitefly infestation alone. Despite the fact that our treatments failed to synergise, our methods can be optimised further, and the effectiveness of the standalone treatments is promising for future studies. In particular, our repellent limonene dispensers were extremely effective at deterring whiteflies and offer a low economic cost and easy to implement whitefly control option. The methods we have used here could be incorporated into current integrated pest management (IPM) systems, a sustainable approach to pest control which will be central to our efforts to manage whitefly populations under glass in the future.

Trap Assays of the Walnut Twig Beetle, Pityophthorus juglandis Blackman (Coleoptera: Curculionidae: Scolytinae), Reveal an Effective Semiochemical Repellent Combination.

Thousand cankers disease (TCD), is an invasive insect-disease complex caused by the walnut twig beetle, Pityophthorus juglandis, and fungal pathogen, Geosmithia morbida. Semiochemical interruption is a viable option for protecting walnut trees from P. juglandis attack. The goal of this study was to test beetle responses to potential repellent compounds. The results of five, flight-intercept assays are reported. Assays 1-3 tested four compounds at variable release rates: (S)-(-)-verbenone, (R)-(+)-verbenone, racemic chalcogran, and racemic trans-conophthorin. Trapping results indicated that the highest release rate tested for each compound was the most effective in reducing the number of beetles caught. (S)-(-)-Verbenone was the least effective, reducing P. juglandis trap catches by 66%. (R)-(+)-Verbenone reduced the number of P. juglandis by 84%. Neither enantiomer of verbenone performed as well as chalcogran or trans-conophthorin, which both reduced the number of beetles caught by ca. 98%. Following individual assays, the most effective compounds were tested in subtractive-combination assays. Combinations of high release rates for (R)-(+)-verbenone, trans-conophthorin, and two stereoisomers of limonene (tested in a previous study) were tested in two assays. The subtractive-combination assays were inconclusive in that trap catches were similar across all treatments. All combination treatments were highly effective, achieving approximately 99% reduction in the number of beetles caught. Based on the trapping results, commercial availability, and cost of the semiochemicals tested, we conclude that a combination of (R)-(+)-limonene, trans-conophthorin, and (R)-(+)-verbenone constitutes an effective tool for reducing P. juglandis trap catches.

Constitutive and Inducible Resistance to Thrips Do Not Correlate With Differences in Trichome Density or Enzymatic-Related Defenses in Chrysanthemum.

Western flower thrips (WFT), Frankliniella occidentalis, is a serious insect pest of Chrysanthemum [Chrysanthemum × morifolium Ramat. (Asteraceae)]. Here we have investigated whether genotypic variation in constitutive and inducible resistance to WFT correlates with phenotypic differences in leaf trichome density and the activity of the defense-related enzyme polyphenol oxidase (PPO) in chrysanthemum. Non-glandular and glandular leaf trichome densities significantly varied among ninety-five chrysanthemum cultivars. Additional analyses in a subset of these cultivars, differing in leaf trichome density, revealed significant variation in PPO activities and resistance to WFT as well. Constitutive levels of trichome densities and PPO activity, however, did not correlate with chrysanthemum resistance to WFT. Further tests showed that exogenous application of the phytohormone jasmonic acid (JA) increased non-glandular trichome densities, PPO activity and chrysanthemum resistance to WFT, and that these effects were cultivar dependent. In addition, no tradeoff between constitutive and inducible resistance to WFT was observed. JA-mediated induction of WFT resistance, however, did not correlate with changes in leaf trichome densities nor PPO activity levels. Taken together, our results suggest that chrysanthemum can display both high levels of constitutive and inducible resistance to WFT, and that leaf trichome density and PPO activity may not play a relevant role in chrysanthemum defenses against WFT.

Repellent, oviposition-deterrent, and insecticidal activity of the fungal pathogen Colletotrichum fioriniae on Drosophila suzukii (Diptera: Drosophilidae) in highbush blueberries.

Spotted-wing drosophila, Drosophila suzukii, and the anthracnose pathogen Colletotrichum fioriniae are an important insect pest and fungal disease of highbush blueberries, respectively, in the United States. However, whether C. fioriniae infection affects D. suzukii preference and performance remains unknown. Here, we conducted choice and no-choice studies to determine the repellent, oviposition-deterrent, and insecticidal effects of C. fioriniae on D. suzukii. In choice tests, blueberry fruit treated with anthracnose solutions containing spores from either field-collected infected fruit ('fruit') or a laboratory C. fioriniae culture ('colony') were less attractive to sexually mature D. suzukii females, but not males, than untreated fruit. The plant tissue (fruit or leaves) did not influence C. fioriniae repellency effects on D. suzukii. In no-choice tests, 55% fewer numbers of eggs were laid on, and 65% fewer adults emerged from, blueberry fruit treated with either the 'fruit' or 'colony' anthracnose solution than untreated fruit. Egg-to-adult D. suzukii survival was also 12% lower on C. fioriniae-infected fruit. No repellency or negative effects on survival were observed when C. fioriniae spores were filtered out of the solution. These findings will help efforts towards the discovery of microbial-derived repellent/oviposition-deterrent compounds that could be used in behavior-based management strategies for D. suzukii.

Exposure to N,N-Diethyl-Meta-Toluamide Insect Repellent and Human Health Markers: Population Based Estimates from the National Health and Nutrition Examination Survey.

N,N-diethyl-meta-toluamide (DEET) is one of the most commonly used insect repellants in the United States, yet the existing literature regarding DEET's potential deleterious impact on humans is mixed and is based mostly on case reports. The primary aim of this study was to address this lack of population-based evidence of the effects of DEET exposure on human health in the United States. Our primary outcome measures were biomarkers related to systemic inflammation (high sensitivity C-reactive protein), immune function (lymphocyte), liver function (aspartate aminotransferase, alanine aminotransferase, and γ-glutamyl transferace), and kidney function (estimated glomerular filtration rate). We analyzed data from the population-based National Health and Nutrition Examination Survey, 2015-2016, and identified 1,205 patients (age 20+ years) who had DEET metabolite levels recorded at or above detection limits. A Pearson correlation was used to assess the relationship between DEET metabolite, and each biomarker found there was no significant correlation. Thus, there is no evidence that DEET exposure has any impact on the biomarkers identified.

In vitro and in vivo metabolic stability of various fragrance materials and insect repellent in rainbow trout (Oncorhynchus mykiss).

Commercial fragrances consist of several thousand natural and synthetic substances formulated in complex combinations. These ingredients are frequently blended at very low concentrations but they are typically lipophilic and a few of them (e.g., synthetic musks) have been detected in aquatic systems, albeit at low concentrations. Few fragrances have guideline in vivo data on bioaccumulation, so in silico modeling has been widely used to estimate bioconcentration factors (BCFs). This study used in vitro metabolism assays with trout S9 cell fractions and cryopreserved hepatocytes to improve estimates of fish BCFs and to test published methods for extrapolating in vitro metabolic rate data to whole fish and corresponding BCFs. These estimates for several chemicals were compared with new in vivo bioconcentration measurements and previously published data on fragrances and the insect repellent, DEET. In total, 17 (20 including isomers) fragrance chemicals (abalyn, amberwood, amboryl acetate, bisabolene, cedroxide, coniferan, elemol, givescone, maritima, precyclemone B, polysantol, sandela, sanjinol, santalex, timberol and vernaldehyde) and DEET were metabolized at various rates. Only three materials tested did not appear to undergo enzymatic degradation (caryophyllene oxide, galaxolide and ketone patchouli). Even relatively slow rates of metabolism had a large influence on bioconcentration estimates. This work adds valuable information to the evolving body of work supporting the use of in vitro determinations of hepatic clearance to improve modeled predictions of bioaccumulation. It can also be used directly to help prioritize testing of potential bioaccumulative chemicals or serve as a more economical method for screening these chemicals.

The role of chemosensory protein 10 in the detection of behaviorally active compounds in brown planthopper, Nilaparvata lugens.

Chemosensory proteins (CSPs) play important roles in insects' chemoreception, although their specific functional roles have not been fully elucidated. In this study, we conducted the developmental expression patterns and competitive binding assay as well as knock-down assay by RNA interference both in vitro and in vivo to reveal the function of NlugCSP10 from the brown planthopper (BPH), Nilaparvata lugens (Stål), a major pest in rice plants. The results showed that NlugCSP10 messenger RNA was significantly higher in males than in females and correlated to gender, development and wing forms. The fluorescence binding assays revealed that NlugCSP10 exhibited the highest binding affinity with cis-3-hexenyl acetate, eicosane, and (+)-ß-pinene. Behavioral assay revealed that eicosane displayed attractant activity, while cis-3-hexenyl acetate, similar to (+)-ß-pinene significantly repelled N. lugens adults. Silencing of NlugCSP10, which is responsible for cis-3-hexenyl acetate binding, significantly disrupted cis-3-hexenyl acetate communication. Overall, findings of the present study showed that NlugCSP10 could selectively interrelate with numerous volatiles emitted from host plants and these ligands could be designated to develop slow-release mediators that attract/repel N. lugens and subsequently improve the exploration of plans to control this insect pest.

Activation of the ambusher tick Rhipicephalus microplus (Acari: Ixodidae) exposed to different stimuli.

The present study aimed to evaluate the behaviour of larvae of Rhipicephalus microplus exposed to different stimuli. A Y-olfactometer was positioned vertically and R. microplus larvae were exposed to environmental air, CO2 alone, N,N-diethyl-3-methylbenzamide (DEET) alone, and CO2 combined with the repellents DEET and (E)-2-octenal. Tests were also conducted with the olfactometer positioned horizontally; in this case, however, only CO2 was tested. In all tests conducted with the Y-olfactometer positioned vertically, CO2 activated R. microplus larvae even in the presence of DEET and (E)-2-octenal, although activation was lower when these repellents were used. In the absence of CO2 , larval behaviour against DEET was similar to that of the larvae in the control group. In the tests performed with the olfactometer positioned horizontally, the larvae had no significant response to the presence of CO2 . The larvae were not attracted to or repelled by any compound tested in either the vertical or horizontal position of the olfactometer. The lack of horizontal displacement, attraction or repellence may have been a result of the ambush behaviour of this tick species. However, when larvae were exposed to stimuli and the olfactometer was positioned vertically, the interference of attractant and repellent stimuli in larval behaviour was assessed.

Liquid Mosquito Repellent Ingestion in Children.

OBJECTIVES: Hydrocarbons are a common cause of accidental poisoning in children, with kerosene being the most implicated agent in rural parts of India. However, lately, liquid mosquito repellent ingestion is emerging as an important cause of hydrocarbon (kerosene) poisoning in urban households. METHODS: This is a retrospective case series over a 5-y period (January 2013 - December 2017) of children with accidental liquid mosquito repellent ingestion presenting to the pediatric emergency. Epidemiology, clinical profile, management and outcomes are discussed. RESULTS: Twenty-three children with median (IQR) age of 24 (18.8-32) mo presented after mean (SD) interval of 6 (3) h from ingestion. Majority (20, 87%) were seen during summer months (March-June) and all were from urban background. Sixteen (70%) had mild-moderate acute respiratory distress syndrome (ARDS) requiring supplemental oxygen with or without positive airway pressure for a mean (SD) duration of 3.3 (1.9) d. All except one survived. CONCLUSIONS: Children with accidental liquid mosquito repellent ingestion had predominant aspiration pneumonitis due to hydrocarbon content rather than neurological complications attributable to synthetic pyrethroids. Ensuring child-proof containers, appropriate storage, regulatory surveillance and parental awareness are must for prevention.