Interference of benomyl and methyl-2-benzimidazolecarbamate (MBC) with DNA metabolism and nuclear divisions in Fusarium oxysporum

Methyl-1-(butylcarbamoyl)-2-benzimidazolecarbamate (benomyl) severely decreased DNA synthesis when applied at 3.5 × 10^?6M during the G1 phase of germinating conidia of Fusarium oxysporum; nuclear divisions were completely inhibited at a fungicide concentration of 10 × 10^?6M. The same concentration applied only after the S phase also completely inhibited the nuclear divisions. This dual interference of benomyl with DNA formation and mitosis might be related to a disturbed phosphorus metabolism.     

Dynamics of uptake,translocation, and disappearance of thiabendazole and methyl-2-benzimidazolecarbamate in pepper and tomato plants

The dynamics of the uptake, translocation, and disappearance of thiabendazole (TBZ) and methyl-2-benzimidazolecarbamate (MBC), the fungicidal breakdown product of benomyl, were studied in tomato and pepper plants grown on nutrient solutions containing 0.5 ppm TBZ and 2.5 ppm MBC. Chemical analysis of pepper plants showed that the fungicides accumulated in the leaves only, where concentrations of 20 ppm TBZ and 60 ppm MBC were recorded. Thiabendazole was completely removed from pepper plants by acetone extraction, whereas MBC was only partially removed by acetone and the rest was weakly bound to the tissue and released by either methanolic HCl extraction or acid hydrolysis. The rate of disappearance of TBZ from pepper leaves was three to four times faster than that of MBC. Balance studies in tomatoes have shown an average disappearance rate of 13.5% per 10 days for MBC. 2-Aminobenzimidazole, the degradation product of MBC, was always detected but its concentration did not exceed 2% of that of the parent compound.     

Sensitivity of Ustilago maydis to pyrazophos and one of its conversion products,and failure to induce resistance with UV-treatment

2-Hydroxy-5-methyl-6-ethoxycarbonylpyrazolo(1,5-a)pyrimidine (PP), a conversion product of pyrazophos, shows considerable toxicity toUstilago maydis, when administered to this fungus in a solution at pH<5. Evidence was obtained thatU. maydis may convert pyrazophos in to PP, and that the latter compound is the toxic principle responsible for the action of pyrazophos. By UV-irradiation of sporidia no PP-resistant mutants were obtained. This does not support the hypothesis that this toxicant acts by specific inhibition of one enzyme system.     

A mutant of Ustilago maydis deficient in sterol C-14 demethylation: Characteristics and sensitivity to inhibitors of ergosterol biosynthesis

An ergosterol-deficient mutant of Ustilago maydis was compared to the wild type in regard to morphology, growth rate, lipid content, and sensitivity to ergosterol biosynthetic inhibitors. Morphology of mutant sporidia is abnormal and resembles that of fenarimol-treated wild-type sporidia. Doubling time of mutant sporidia is 6.3 hr compared to 2.5 hr for the wild type. The mutant produces 24-methylenedihydrolanosterol, obtusifoliol, and 14α-methylfecosterol; ergosterol is absent. The sterols of the mutant are the same as those which accumulate in wild-type sporidia treated with the sterol C-14 demethylation inhibitors fenarimol, etaconazole, and miconazole. The level of free fatty acids is higher in the mutant than in wild-type cells. Growth of mutant sporidia is not inhibited by fenarimol, etaconazole, and miconazole, or by the sterol Δ¹⁴-reductase inhibitor azasterol A25822B at low concentrations which inhibit growth of wild-type sporidia. The residual growth rate of wild-type sporidia treated with low concentrations of the sterol C-14 demethylation inhibitors is about the same as that of untreated mutant sporidia. Therefore, the mutant would not be recognized as resistant in a wild-type population. The mutant is deficient in sterol C-14 demethylation and is similar in all properties studied to wild-type sporidia treated with sterol C-14 demethylation inhibitors. These findings support the contention that inhibition of sterol C-14 demethylation in U. maydis is the primary mode of toxicity of fenarimol, etaconazole, and miconazole. A secondary mode of toxicity is evident for miconazole and etaconazole at higher concentrations but is doubtful for fenarimol.     

The mechanism of resistance to sterol 14α demethylation inhibitors in a mutant (Erg 40) of Ustilago maydis

Resistance to DMI fungicides is a problem in both agriculture and medicine. Several mechanisms of resistance exist, but, as yet, few have been characterised in field resistant strains of plant pathogens. One approach to evaluating the role of mutations in the sterol 14α demethylase (14DM) target site requires cloning this gene and confirming its identity by complementation in an appropriate mutant. The azole‐resistant mutant, Erg 40, of Ustilago maydis which is totally blocked at the 14α demethylation step in sterol biosynthesis seems to be suitable for such expression studies. Transformation of Erg 40 with a plasmid containing the yeast 14α demethylase (CYP51A1) gene removed the block in sterol biosynthesis and generated azole‐sensitive transformants. Detailed analysis of these transformants failed to detect the presence of the yeast gene and suggested, instead, that changes in sterol biosynthesis resulted simply from the transformation protocol and not from the incorporation of extracellular DNA. Subsequent sequence analysis has revealed a mutation in the 14α demethylase gene of Erg 40. The results suggest that azole resistance in Erg 40 is not simply controlled by this mutation but involves some additional regulatory function, and consequently Erg 40 is not suitable for complementation studies with CYP51A1 genes. © 2000 Society of Chemical Industry     

Inhibition of ergosterol biosynthesis in Ustilago maydis by the fungicide 1-[2-(2, 4-dichlorophenyl)-4-ethyl-1, 3-dioxolan-2-ylmethyl]-1H-1, 2, 4-triazole

Inhibition of sporidial multiplication in cultures of Ustilago maydis by 1-[2-(2, 4-dichlorophenyl)-4-ethyl-1, 3-dioxolan-2-ylmethyl]-1H-1, 2, 4-triazole^a (CGA-64251), at concentrations of 0.1, 1.0 and 5.0 μg ml^?1, increased from about 15% during the first 4 h, to 58–70% during the subsequent 4 to 12-h period. Sporidia became swollen and highly branched in the presence of the fungicide. Total lipid content as a percentage of the dry weight was not affected after exposure of the sporidia to the fungicide at 0.1 or 5 μg ml^?1 for 4 h, but synthesis of ergosterol and other demethyl-sterols was inhibited by 87–92%. Large quantities of methyl-sterol precursors of ergosterol and of free fatty acids accumulated in the treated sporidia. Fungitoxicity of CGA-64251 is attributed to inhibition of ergosterol biosynthesis at the stage of sterol C-14 demethylation.     

Comparison of solvents and methods of extracting methylbenzimidazol-2-yicarbamate (MBC) from tobacco

The bioautographic method of Homans and Fuchs for detecting MBC on thin layer plates, was tested quantitatively. The sum of the vertical and horizontal diameters of the zones of inhibition of Penicillium expansum, and the square root of the mass of an equal area of paper, were directly proportional to the logarithm of the concentration of MBC. Methanol and ethanol extracted significantly more MBC from tobacco tissue than acetone and chloroform. Soxhlet extraction was more effective than cold (22 °C) treatment, but the latter, in which 100 mg aliquots of dried tissue were used, was the least variable, very much quicker and could be used to detect the fungicide in small amounts of individual plant tissues.     

Baseline toxicity of several pesticides to Hyaliodes vitripennis (Say) (Hemiptera: Miridae)

Hyaliodes vitripennis (Say) is a univoltine indigenous predacious mirid. It has been reported in several orchards where IPM programmes are used. It is a generalist, and feeds on phytophagous mites in addition to other arthropods. In Quebec, a foliar application of imidacloprid, deltamethrin or lambda‐cyhalothrin is used at least once per season to manage arthropod pests such as leafhoppers and leaf‐eating caterpillars. Meanwhile, several applications of metiram, flusilazole, myclobutanil and mancozeb are made to control apple scab [Venturia inaequalis (Cooke) Winter]. In laboratory trials, comparison of lethal concentrations of the three insecticides against H vitripennis nymphs and adults showed no significant difference. However, when lethal concentrations were compared between two growth stages for each insecticide, a significant difference was noted between adults and nymphs treated with lambda‐cyhalothrin, adults being more susceptible than nymphs. No such difference could be detected for imidacloprid or deltamethrin. When LC₅₀ values were compared with the manufacturer's label rates, deltamethrin and imidacloprid were toxic to the nymphs and adults, and lambda‐cyhalothrin was slightly toxic to the nymphs and moderately toxic to the adults. Among the fungicides evaluated in the laboratory, myclobutanil showed moderate toxicity to adults at the manufacturer's label rate. The remaining fungicides had no toxic effects to adults or nymphs, even at four times the manufacturer's label rate. © 2001 Society of Chemical Industry     

Evaluation of semiochemical toxicity to houseflies and stable flies (Diptera: Muscidae)

BACKGROUND: The housefly, Musca domestica L., and stable fly, Stomoxys calcitrans (L.) are cosmopolitan pests of both farm and home environments. Houseflies have been shown to be resistant to a variety of insecticides, and new chemistries are slow to emerge on the market. Toxicities of selected semiochemicals with molecular structures indicative of insecticidal activity were determined against adults from an insecticide‐susceptible laboratory strain of houseflies. The three most active semiochemicals were also evaluated against recently colonized housefly and stable fly strains. RESULTS: Nineteen semiochemicals classified as aliphatic alcohols, terpenoids, ketones and carboxylic esters showed toxicity to houseflies and stable flies. Rosalva (LC₅₀ = 25.98 µg cm^?2) followed by geranyl acetone and citronellol (LC₅₀ = 49.97 and 50.02 µg cm^?2) were identified as the most toxic compounds to houseflies. Permethrin was up to 144‐fold more toxic than rosalva on the susceptible strain. However, it was only 35‐fold more toxic to the insecticide‐tolerant field strain. The compounds generated high toxicity to stable flies, with LC₅₀ values ranging from 16.30 to 40.41 µg cm^?2. CONCLUSION: Quantification of LC₅₀ values of rosalva, citronellol and geranyl acetone against susceptible housefly and field‐collected housefly and stable fly strains showed that semiochemicals could serve as potent insecticides for fly control programs. Copyright © 2010 Society of Chemical Industry     

Acute and sublethal toxicity of novaluron, a novel chitin synthesis inhibitor, to Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)

The acute and sublethal toxicities of novaluron, a novel chitin synthesis inhibitor, to a laboratory-reared insecticide-susceptible strain of Colorado potato beetle, Leptinotarsa decemlineata (Say), were determined. Novaluron exhibited excellent residual (120 h LC(50) = 0.42 mg litre(-1)) and good direct contact (120 h LC(50) = 27 mg litre(-1)) activity against second-instar larvae (L2). Hatch of eggs exposed by direct contact to novaluron solutions > or =100 mg litre(-1) was significantly reduced, as was the ability of emerged first-instar larvae to moult. L2 from eggs exposed to > or =100 mg litre(-1) novaluron weighed significantly less (P < 0.0001) than those from untreated eggs. However, L2 from eggs treated with 1 mg litre(-1) novaluron weighed significantly more (P < or = 0.05) than those from untreated eggs, suggesting novaluron can have a hormetic effect on L decemlineata larval development. Leptinotarsa decemlineata mating pairs fed foliage treated with novaluron at 25 or 75 g AI ha(-1) produced approximately 25% fewer egg masses and eggs per mass. Hatch of eggs on treated foliage was almost completely suppressed, and longevity of male beetles was reduced by approximately 50% when fed foliage treated with novaluron at 75 g AI ha(-1).     

Evaluation of semiochemical toxicity to Aedes aegypti,Ae. albopictus and Anopheles quadrimaculatus (Diptera: Culicidae)

BACKGROUND: Mosquitoes are the most important vectors of human pathogens. Wide‐scale use of pesticides has led to the development of resistance to most common insecticide groups. The need to develop novel products that have a low impact on human health and the environment is well established. The toxicity of selected semiochemicals with molecular structures indicative of insecticidal activity was determined against adult Aedes aegypti (L.) and Anopheles quadrimaculatus (Say). The two most active insecticides against Ae. aegypti were also evaluated against Ae. albopictus (Skuse). RESULTS: Fifteen semiochemicals classified as terpenoid alcohols, ketones or carboxylic esters showed toxicity to both mosquito species. Geranyl acetone (LC₅₀ = 38.51 µg cm^?2) followed by citronellol (LC₅₀ = 48.55 µg cm^?2) were the most toxic compounds to Ae. aegypti, while geraniol and lavonax, with LC₅₀ values of 31.88 and 43.40 µg cm^?2, showed the highest toxicity to An. quadrimaculatus. Both geranyl acetone and citronellol were highly toxic to Ae. albopioctus. No semiochemical showed fumigation activity against either species. All semiochemicals persisted for less than 24 h when tested on filter paper. CONCLUSION: Quantification of LC₅₀ values of several semiochemicals against Ae. Aegypti, An. quadrimaculatus and Ae. albopioctus showed that semiochemicals not only modify insect behaviors but also hold potential as potent insecticides for mosquito control programs. Copyright © 2010 Society of Chemical Industry     

Biochemical and histopathological studies to assess chronic toxicity of triazophos in blood, liver and brain tissue of rats

Triazophos, O,O-diethyl-1-H-1,2,4-triazol-3-yl phosphorothioate, (TZ) is an organophosphorus pesticide which is extensively used in agriculture for controlling insect pests. Except a FAO/WHO report no study has investigated its short-term toxicity with respect to its potential to cause biochemical and histopathological alterations. The present study was designed to identify the effect of TZ at different doses (1.64, 3.2 and 8.2 mg/kg) on the oxidative stress parameters in blood as well as organs involved in xenobiotic metabolism (liver and brain) following chronic exposure for 90 days. Moreover, the study also delineates the effect of TZ on the histo-architecture of these organs. The results indicated a dose dependent induction (p < 0.001) of oxidative stress, as evident by increased malondialdehyde (MDA) level and compromised antioxidant defense including glutathione S transferase (GST) activity, glutathione (GSH) content and ferric reducing ability of plasma (FRAP) in blood, and increased MDA level with concomitantly decreased GSH content in tissues, following chronic exposure to TZ. The ratio of MDA: FRAP in blood was found to be increased following chronic exposure to TZ and may serve as a suitable indicator of severity of oxidative damage. Onset of such biochemical alterations is one of the early adaptive responses to TZ exposure which leads to histopathological alterations in terms of diffuse fatty changes expanding from mid-zonal area to whole lobule in liver. However, increased oxidative stress did not bring any morphological alteration in brain. The present study concludes that induction of oxidative stress, leading to subsequent histopathological alterations in liver, is an important mechanism underlying the TZ induced chronic toxicity.     

Inhibition of two acetylcholinesterases by the 4-nitrophenyl esters of methyl-, ethyl-, and isopropyl(phenyl)phosphinic acid

The inhibition of eel acetylcholinesterase and bovine erythrocyte acetylcholinesterase by the 4-nitrophenyl esters of methyl-, ethyl-, and isopropyl(phenyl)phosphinic acid (MPP, EPP, and IPP, respectively) was investigated at pH 6.90 in 0.067 M phosphate buffer (25.0°C) using stopped-flow instrumentation and automated data processing. Our evaluation of the dissociation constant, K_d, the unimolecular bonding rate constant, k₂, and the bimolecular reaction constant, k_i, are the first reported values for these constants for a homologous series of this class of organophosphorus compounds. The largest k₁ value (29,428 M^?1 sec^?1) was observed for the reaction of eel acetylcholinesterase with 4-nitrophenyl methyl(phenyl)phosphinate. The smallest k_i value (9.6 M^?1 sec^?1) was observed for the reaction of bovine erythrocyte acetylcholinesterase with 4-nitrophenyl isopropyl(phenyl)phosphinate.     

The toxicity of cyromazine to Chironomus zealandicus (chironomidae) and Deleatidium sp. (leptophlebiidae)

The toxicity of cyromazine and a commercial formulation, ‘Vetrazin’®, to Chironomus zealandicus (thummi) Hudson and Deleatidium sp. was investigated. Under acute test conditions, the LC₅₀ values for each species were quite comparable. For C. zealandicus, the value varied according to instar, 100–400 mg litre^?1 for second- and third-instar to 1000–10000 mg litre^?1 for older fourth-instars. For the one size class of Deleatidium tested (c.10 mm long), the value was 300–400 mg litre^?1. High control mortalities of C. zealandicus limit that species' usefulness as an acute bioassay candidate. Under chronic test conditions, cyromazine showed a high toxicity to eggs or early-instar larvae of C. zealandicus. The maximum acceptable toxicant concentration for cyromazine against C. zealandicus was approximately 17.5 μg litre^?1. The possibility of water contamination at this level is discussed. Whole-of-life chronic tests with C. zealandicus indicated that the most susceptible stage was in the egg or soon after larval emergence. These results highlight the dangers of using short-term acute toxicity results to formulate environmental exposure limits for modern pesticides that do not have dysfunction of the nervous system as their mode of action.     

氯虫苯甲酰胺对2种跳虫的繁殖毒性与氧化胁迫

为研究氯虫苯甲酰胺 (chlorantraniliprole, CAP) 对跳虫的繁殖毒性和氧化胁迫效应,以2种跳虫白符跳Folsomia candida Willem和奇裸长跳Sinella insolens为受试生物,分别测定了CAP对其28 d和21 d的繁殖毒性和暴露于CAP亚致死剂量（白符跳0.0533 mg/kg,奇裸长跳100 mg/kg）10 d内,2种跳虫体内抗氧化防御系统受到的影响。结果表明,CAP对白符跳和奇裸长跳的繁殖率半抑制浓度 (EC_50-repro) 分别为0.533 mg/kg dw (95%置信区间为0.370~0.769 mg/kg dw) 和 > 1 000 mg/kg dw,毒性差异明显。暴露于亚致死剂量的CAP 0、2、4、6和10 d后,CAP能不同程度地影响2种跳虫的抗氧化防御系统。暴露初期过氧化氢酶 (CAT) 活性分别上升113%和108%,谷胱甘肽还原酶 (GR) 活性分别上升141%和74.6% (P < 0.05),随暴露时间延长,2种酶活性逐渐下降,最终趋向对照组水平;总谷胱甘肽 (TG) 活性水平始终维持在对照组以上,并在第6天达到最高;谷胱甘肽-S-转移酶 (GST) 活性在暴露初期分别下降38.4%和21.6% (P < 0.05),随暴露时间延长逐渐升高,回到对照组水平;脂质过氧化物 (LPO) 含量随暴露时间延长而呈现先上升后下降的趋势,变化幅度较小;而金属硫蛋白 (MT) 含量则未随暴露时间的延长而出现显著变化。本研究结果表明,白符跳对CAP更为敏感,可作为土壤环境中CAP的指示生物。CAT、GR和GST是指示CAP短期胁迫的较适宜的生物标志物,而TG和LPO可以作为指示CAP中长期胁迫的生物标志物。     

Biochemical characterization of chlorantraniliprole and spinetoram resistance in laboratory-selected obliquebanded leafroller, Choristoneura rosaceana (Harris) (Lepidoptera: Tortricidae)

Neonate larvae of obliquebanded leafroller, Choristoneura rosaceana, from a laboratory colony were exposed to two reduced-risk insecticides, chlorantraniliprole and spinetoram. After nine generations of selection, significant levels of resistance to each insecticide were observed. Biochemical assays were performed on third instars to determine potential resistance mechanisms. Enzyme assays indicated that esterase activity was significantly increased in the chlorantraniliprole-selected colony, whereas mixed-function oxidase levels were elevated in the spinetoram-selected colony as compared to the unselected colony. No difference in glutathione-S-transferase activity was seen in either of the insecticide-selected colonies. These results indicate the potential involvement of esterases and mixed-function oxidases as detoxification mechanisms responsible for resistance to chlorantraniliprole and spinetoram, respectively. Furthermore, the results of this study suggest that chlorantraniliprole and spinetoram are not detoxified by similar mechanisms and could therefore be incorporated into resistance management programs in tree fruit leading to sustainable management of C. rosaceana.