Residue analysis and dietary exposure risk assessment of tebufenozide in stem lettuce (Lactuca sativa L. var. angustana Irish).

Tebufenozide, a newly-developed nonsteroidal ecdysone agonist, is in pre-regulation phase (before approval for use) on stem lettuce in China. Aiming at the safe application of tebufenozide, the dissipation and terminal residue trials on stem lettuce were performed under good agricultural practice (GAP). The dissipation trials shown that tebufenozide was rapidly degraded in stem lettuce, with half-lives of 5.0-8.2 days. Pre-regulation dietary exposure risk assessments were evaluated to recommend maximum residue limits (MRLs) based on risk quotients (RQ) method. Relevant toxicological parameters including ADI (acceptable daily intake) and ARfD (acute reference dose) were applied to assess the potential dietary exposure risk. The results indicated the chronic dietary exposure risk probability (RQc) of tebufenozide ranged from 36.4% to 70.0%. The acute dietary exposure risk probability (RQa) of tebufenozide was 2.88%-8.49% in lettuce stems and 14.0%-20.0% in lettuce leaves, respectively. On the basis of supervised field trial data and dietary exposure risk assessment results, the MRLs of tebufenozide were recommended as 3 mg/kg for lettuce stems and 10 mg/kg for lettuce leaves, respectively. The results demonstrated that the dietary exposure risk of tebufenozide used in stem lettuce under GAP was negligible and would not pose unacceptable health risk to Chinese consumers.

Cyromazine resistance in a field strain of house flies, Musca domestica L.: Resistance risk assessment and bio-chemical mechanism.

Developing resistance management strategies for eco-friendly insecticides is essential for the management of insect pests without harming the environment. Cyromazine is a biorational insecticide with very low mammalian toxicity. Resistance to cyromazine has recently been reported in house flies from Punjab, Pakistan. In order to propose a resistance management strategy for cyromazine, experiments were planned to study risk for resistance development, possibility of cross-resistance and bio-chemical mechanisms. A field strain of house flies with 8.78 fold resistance ratio (RR) to cyromazine was re-selected under laboratory conditions. After seven rounds of selection (G1-G7), the RR values rapidly increased from 8.8 to 211 fold. However, these values declined to 81fold when the cyromazine selected (CYR-SEL) strain was reared without selection pressure, suggesting an unstable nature of resistance. The CYR-SEL strain showed lack of cross-resistance to pyriproxyfen, diflubenzuron, and methoxyfenozide. Synergism bioassays using enzyme inhibitors: piperonyl butoxide (PBO) and S,S,S-tributylphosphorotrithioate (DEF), and metabolic enzyme analyses revealed increased activity of carboxylesterase (CarE) and mixed-function oxidase (MFO) in the CYR-SEL strain compared to the laboratory susceptible (Lab-susceptible) strain, suggesting the metabolic resistance mechanism responsible for cyromazine resistance in the CYR-SEL strain. In conclusion, risk of rapid development of cyromazine resistance under consistent selection pressure discourages the sole reliance on cyromazine for controlling house flies in the field. The unstable nature of cyromazine resistance provides window for restoring cyromazine susceptibility by uplifting selection pressure in the field. Moreover, lack of cross-resistance between cyromazine and pyriproxyfen, diflubenzuron, or methoxyfenozide in the CYR-SEL strain suggest that cyromazine could be rotated with these insecticides whenever resistance crisis occur in the field.

Epigenetic Regulation in Environmental Chemical Carcinogenesis and its Applicability in Human Health Risk Assessment.

Although several studies have shown that chemically mediated epigenetic changes are an etiological factor in several human disease conditions, the utility of epigenetic data, such as DNA methylation, in the current human health risk assessment paradigm is unclear. The objective of this study is to investigate the relationship between the points of departure (PODs) for cancer incidence and DNA methylation changes in laboratory animals exposed to the following environmental toxicants: bromodichloromethane, dibromochloromethane, chloroform, hydrazine, trichloroethylene, benzidine, trichloroacetic acid, and di(2-ethylhexyl) phthalate (DEHP; a known reproductive toxicant). The results demonstrate that the PODs for cancer incidence and altered DNA methylation are similar. Furthermore, based on the available data, the POD for DNA methylation appeared more sensitive compared to that for cancer incidence following the administration of DEHP to rats during different life stages. The high degree of correlation between PODs for cancer incidence and DNA methylation (for both total DNA and individual genes) suggests that DNA methylation end points could potentially be used as a screening tool in predicting the potential toxicity/carcinogenicity and in prioritizing large numbers of chemicals with sparse toxicity databases. The life stage during which treatment occurs is also an important consideration when assessing the potential application of epigenetic end points as a screening tool.

Fitness Cost of Methoxyfenozide and the Effects of Its Sublethal Doses on Development, Reproduction, and Survival of Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae).

In the present study, two experiments were designed to investigate the lethal and sublethal effects of methoxyfenozide on Spodoptera litura (F.). The first experiment was conducted to explore the fitness cost of methoxyfenozide resistance in a methoxyfenozide-resistant strain (selected with methoxyfenozide for 13 consecutive generations), leading to resistance ratio to methoxyfenozide 83.0- and 2359-fold higher when compared to the field and susceptible populations with a fitness cost of 0.17. In the second experiment, second instars of the susceptible strain were treated with sublethal doses of methoxyfenozide (LC30, LC20, and LC10) by diet incorporation for larval feeding for 3 days. It was observed that higher concentrations of methoxyfenozide significantly prolonged the larval and pupal development time of S. litura as compared to the control treatment. The number of eggs per female, egg hatching, sex ratio, and longevity of adults of methoxyfenozide-treated groups was greatly reduced as compared to untreated S. litura. Our data clearly indicated that fitness cost of methoxyfenozide and its sublethal effects on S. litura has an important impact on its population dynamics.

Assessment of species specificity of moulting accelerating compounds in Lepidoptera: comparison of activity between Bombyx mori and Spodoptera littoralis by in vitro reporter and in vivo toxicity assays.

BACKGROUND: Dibenzoylhydrazine analogues have been developed successfully as a new group of insect growth regulators, called ecdysone agonists or moulting accelerating compounds. A notable feature is their high activity against lepidopteran insects, raising the question as to whether species-specific analogues can be isolated. In this study, the specificity of ecdysone agonists was addressed through a comparative analysis in two important lepidopterans, the silkworm Bombyx mori L. and the cotton leafworm Spodoptera littoralis (Boisd.). RESULTS: When collections of non-steroidal ecdysone agonists containing different mother structures (dibenzoylhydrazine, acylaminoketone, tetrahydroquinoline) were tested, in vitro reporter assays showed minor differences using cell lines derived from both species. However, when compounds with high ecdysone agonist activity were examined in toxicity assays, larvicidal activity differed considerably. Of note was the identification of three dibenzoylhydrazine analogues with > 100-fold higher activity against Bombyx than against Spodoptera larvae. CONCLUSION: The present study demonstrated that species-specific ecdysone-agonist-based insecticides can be developed, but their species specificity is not based on differences in the activation of the ecdysone receptor but rather on unidentified in vivo parameters such as permeability of the cuticle, uptake/excretion by the gut or metabolic detoxification.

Hepatic gene expression and lipid homeostasis in C57BL/6 mice exposed to hydrazine or acetylhydrazine.

Hydrazine (HD) and acetylhydrazine (AcHD) are metabolites of the antituberculosis drug isoniazid (INH) that have been implicated in INH-induced liver damage. The hepatotoxicity of AcHD and HD were compared in adult male C57Bl/6J mice by evaluating hepatic histopathology, plasma biochemistry, and hepatic gene expression. By all measures, HD had significantly greater effects than AcHD. There was no evidence of liver damage following exposure to AcHD (300 mg/kg, po). However, HD at this dose caused marked hepatic necrosis, macrovesicular degeneration, and steatosis. Lipid accumulation was initiated 2 h after HD exposure, with hepatic macrovesicular degeneration evident after 4 h, and severe necrosis by 36 h. Gene expression profiles were compared 24 h following 100 mg/kg po of HD or AcHD. HD changed the hepatic expression of more genes than AcHD, particularly lipid synthesis, transport, and metabolism genes that may be involved in steatosis. Hepatic expression of genes regulated by peroxisome proliferator activated receptors (PPAR) and sterol regulatory element binding protein (SREBP) transcription factors was increased only by HD. The hepatotoxicty and hepatic gene expression profile of HD, but not AcHD, indicate that exposure to HD initiates a process whereby the production and intracellular transport of hepatic lipids is favored over the removal of fatty acids and their metabolites.

Risk assessment of high-energy chemicals by in vitro toxicity screening and quantitative structure-activity relationships.

Hydrazine propellants pose a substantial operational concern to the U.S. Air Force and to the aerospace industry because of their toxicity. In our continuing efforts to develop methods for the prediction of the toxicological response to such materials, we have measured in vitro toxicity endpoints for a series of high-energy chemicals (HECs) that were recently proposed as propellants. The HECs considered are structurally diverse and can be classified into four chemical types (hydrazine-based, amino-based, triazoles, and a quaternary ammonium salt), although most are hydrazine derivatives. We measured the following endpoints in primary cultures of isolated rat hepatocytes: mitochondrial function (MTT), lactate dehydrogenase leakage (LDH), generation of reactive oxygen species (ROS), and total glutathione content (GSH). In several instances, effective concentrations (EC) were indeterminate, and only lower limits to the measured endpoints could be ascertained. Using molecular descriptors calculated with a semiempirical molecular orbital method, quantitative structure-activity relationships (QSARs) were derived for MTT (EC25) and for GSH (EC50). Correlation coefficients for 2- and 3-parameter QSARs of about 0.9 enable us to predict the toxicity for similar compounds. Furthermore, except in one case, predicted EC values for the uncertain endpoints were consistent with experiment. Descriptors comprising the QSARs for MTT were consistent with the biophysical mechanism of toxic response found experimentally for hydrazine derivatives. Application of our derived QSARs will assist in predicting toxicity for newly proposed propellants.

In vitro toxicity assessment of a new series of high energy compounds.

Hydrazine is an aircraft fuel and propellant used by the US Air Force. Due to its toxicity the Propulsion Directorate of the Air Force Research Laboratory (AFRL/PR) has investigated alternative chemicals to replace hydrazine. AFRL/PR has synthesized a series of high energy chemicals (HECs), primarily hydrazine derivatives and amino containing compounds such as hydrazinium nitrate (HZN), 2-hydroxyethyl-hydrazine nitrate (HEHN), diethyl hydrazine nitrate (DEHN), ethanolamine nitrate (EAN), histamine dinitrate (HDN) and methoxylamine nitrate (MAN) to study as alternative chemical candidates. Although HECs are reliable constituents of powered propellant systems, they constitute an important class of toxic agents to which military and civilian personnel can be exposed. The current study was undertaken to examine the toxicity of HECs in primary hepatocytes in vitro. The effects of short-term exposure (4 h) of hepatocytes to HECs were investigated with reference to viability, mitochondrial function and oxidative stress markers. The results showed a decrease in mitochondrial activity, increase in lactate dehydrogenase (LDH) leakage and depletion of reduced glutathione (GSH) levels. The levels of reactive oxygen species (ROS) increased dose dependently in HZN, MAN and HDN exposed cells. However, there was no induction of ROS generation in EAN, DEHN and HEHN exposed cells. Depletion of GSH in hepatocytes by buthionine sulfoximine (BSO) prior to exposure to HZN increased its toxicity. The results suggest that at least one mechanism of HEC toxicity is mediated through oxidative stress.

Gyromitra esculenta mushroom: a comparative assessment of its carcinogenic potency.

A comparative assessment is performed on one of the false morel mushrooms, Gyromitra esculenta, including the amounts of some hydrazines present in this fungus, the cancer-inducing doses of these chemicals or the mushroom used in animal experiments, the total amounts of the hydrazines or mushroom needed to induce neoplasms in mice and the estimated total amounts of hydrazines or mushroom needed to induce cancer in humans. When one compares the estimated amounts of hydrazines required to induce cancer with the amount of raw Gyromitra esculenta needed to yield a similar effect, it becomes clear that to date 37 percent of the carcinogenic ingredients of this fungus have been identified.

Toxicity assessment of hydrazine fuels.

The major health aspects of exposure to hydrazine propellants are reviewed. Toxic effects of hydrazine fuels on humans and animals as well as in vitro studies are discussed with emphasis on recent findings and USAF studies. Propellant hydrazines have been found to be genotoxic in in vitro studies and oncogenic in animal studies. Embryotoxicity has been demonstrated at very high exposures but not at occupationally encountered levels for hydrazine and unsymmetrical dimethylhydrazine. Epidemiologic evidence to support these findings is lacking; however, the results of animal and in vitro studies have resulted in lowering both the time-weighted average-threshold limit values and short-term exposure limits for these propellants.