Oxidative stress and locomotor behaviour response as biomarkers for assessing recovery status of mosquito fish, Gambusia affinis after lethal effect of an organophosphate pesticide, monocrotophos

Recovery study was performed at regular intervals to establish the time course of 50% and 100% recovery in neurotransmitter enzyme (acetylcholinesterase, AChE, EC 3.1.1.7) and locomotor behaviour response of mosquito fish, Gambusia affinis exposed to lethal concentration (20.49 mg L⁻¹) of an organophosphorous pesticide, monocrotophos (MCP) for 96 h. In vitro AChE activity studies indicated that MCP could cause 50% inhibition (I₅₀) at 10.2 × 10⁻⁵ M. A positive correlation was observed between brain AChE activity and swimming speed during the recovery study. Also, the recovery response of the antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) as well as lipid peroxidation (LPO) as biomarkers of oxidative stress were assessed in viscera of G. affinis. The results showed that the MCP besides its inhibitory effect on target enzyme AChE activity and induction in antioxidant enzyme activities as a characteristic of oxidative stress, which can be used as biomarkers in the pesticide contaminated aquatic streams.     

Acetylcholinesterase point mutations putatively associated with monocrotophos resistance in the two-spotted spider mite

Molecular mechanisms of monocrotophos resistance in the two-spotted spider mite (TSSM), Tetranychus urticae Koch, were investigated. A monocrotophos-resistant strain (AD) showed ca. 3568- and 47.6-fold resistance compared to a susceptible strain (UD) and a moderately resistant strain (PyriF), respectively. No significant differences in detoxification enzyme activities, except for the cytochrome P450 monooxygenase activity, were found among the three strains. The sensitivity of acetylcholinesterase (AChE) to monocrotophos, however, was 90.6- and 41.9-fold less in AD strain compared to the UD and PyriF strains, respectively, indicating that AChE insensitivity mechanism plays a major role in monocrotophos resistance. When AChE gene (Tuace) sequences were compared, three point mutations (G228S, A391T and F439W) were identified in Tuace from the AD strain that likely contribute to the AChE insensitivity as predicted by structure analysis. Frequencies of the three mutations in field populations were predicted by quantitative sequencing (QS). Correlation analysis between the mutation frequency and actual resistance levels (LC₅₀) of nine field populations suggested that the G228S mutation plays a more crucial role in resistance (r² = 0.712) compared to the F439W mutation (r² = 0.419). When correlated together, however, the correlation coefficient was substantially enhanced (r² = 0.865), indicating that both the F439W and G228S mutations may work synergistically. The A391T mutation was homogeneously present in all field populations examined, suggesting that it may confer a basal level of resistance.     

Acetylcholinesterase in selected plant-parasitic nematodes: Inhibition, kinetic and comparative studies

The in vitro inhibition potency of some organophosphates (OPs) and carbamates (CAs) which are widely used to control plant-parasitic nematodes on acetylcholinesterase (AChE) of Meloidogyne javanica, Heterodera avenae and Tylenchulus semipenetrans, the major pathogens responsible for the damage of a wide range of crops in Al-Qassim region, Saudi Arabia was examined. AChE of H. avenae activity was 1.58- and 1.51-fold greater than that of T. semipenetrans or M. javanica, respectively. The order of inhibition potency of the tested compounds against T. semipenetrans AChE was: carbofuran > paraoxon > oxamyl > fenamiphos > phorate-sulfoxide > aldicarb, where the corresponding concentrations that inhibited 50% of the nematode AChE activity (I₅₀) were 5 × 10⁻⁸, 7 × 10⁻⁷, 7.5 × 10⁻⁷, 2 × 10⁻⁶, 2 × 10⁻⁴ and 2 × 10⁻³ M, respectively. Paraoxon, fenamiphos and carbofuran exhibited high inhibition potency against M. javanica AChE where the I₅₀ values were below 1 nM. Phorate-sulfoxide and aldicarb were potent inhibitors of M. javanica AChE with I₅₀ values of 3.8 and 8 nM, respectively, while oxamyl exhibited low inhibition potency with I₅₀ of 15 nM. Fenamiphos and paraoxon showed the highest I₅₀ values of <100 μM against H. avenae followed by oxamyl (I₅₀ < 1 mM), whereas paraoxon, carbofuran and aldicarb showed low potency with I₅₀ values >1 mM. All the tested compounds exhibited high inhibition potency to AChE of M. javanica than T. semipenetrans or H. avenae. Except phorate-sulfoxide in M. javanica the inhibition pattern and implied mechanism for all the tested compounds for the three nematodes is suggested to be a linear mixed type (a combination of competitive and non-completive type).     

Single and simultaneous exposure of acetylcholinesterase to diazinon, chlorpyrifos and their photodegradation products

In vitro inhibition of electric eel acetylcholinesterase (AChE) by single and simultaneous exposure to organophosphorus insecticides diazinon and chlorpyrifos, and their transformation products, formed due to photoinduced degradation, was investigated. Increasing concentrations of diazinon, chlorpyrifos and their oxidation products, diazoxon and chlorpyrifos-oxon, inhibited AChE in a concentration-dependent manner. IC₅₀ (20 min) values, obtained from the inhibition curves, were (in mol/l): (5.1 ± 0.3) × 10⁻⁸, (4.3 ± 0.2) × 10⁻⁶ and (3.0 ± 0.1) × 10⁻⁸ for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively, while maximal diazinon concentration was lower than its IC₅₀ (20 min). Calculated K_I values, in mol/l, of 7.9 × 10⁻⁷, 9.6 × 10⁻⁶ and 4.3 × 10⁻⁷ were obtained for diazoxon, chlorpyrifos and chlorpyrifos-oxon, respectively. However, 2-isopropyl-4-methyl-6-pyrimidinol (IMP) and 3,5,6-trichloro-2-pyridinol, diazinon and chlorpyrifos hydrolysis products, did not noticeably affect the enzyme activity at all investigated concentrations. Additive inhibition effect was achieved for lower concentrations of the inhibitors (diazinon/diazoxon ?1 × 10⁻⁴/1 × 10⁻⁸ mol/l i.e., chlorpyrifos/chlorpyrifos-oxon ?2 × 10⁻⁶/3 × 10⁻⁸ mol/l), while an antagonistic effect was obtained for all higher concentrations of the organophosphates. Inhibitory power of 1 × 10⁻⁴ mol/l diazinon irradiated samples can be attributed mostly to the formation of diazoxon, while the presence of non-inhibiting photodegradation product IMP did not affect diazinon and diazoxon inhibitory efficiencies.     

Binding of phenthoate to bovine serum albumin and reduced inhibition on acetylcholinesterase

Organophosphorus pesticides (OPs) are of environmental significance due to their high toxicity to animals. Binding to plasma proteins may effective influence the toxicological properties of xenobiotics. In an attempt to evaluate the affinity of phenthoate (PTA) to bovine serum albumin (BSA) and inhibitory ability of bound PTA to acetylcholinesterase (AChE), we investigated the interactions between phenthoate (PTA) and bovine serum albumin (BSA) using tryptophan fluorescence quenching and subsequent inhibition on AChE activity by PTA. The results showed that PTA caused the fluorescence quenching of BSA because of the formation of a PTA-BSA complex. Quenching constants (K_sv), determined using the Sterns-Volmer equation to provide a measure of the binding affinity between PTA and BSA at 303, 306, 310 and 313 K were (3.4295 ± 0.0763) × 10⁻⁴, (3.2446 ± 0.0635) × 10⁻⁴, (3.0434 ± 0.0856) × 10⁻⁴ and (2.8262 ± 0.0569) × 10⁻⁴ M⁻¹, respectively. The thermodynamic parameters, ΔH and ΔS were −25.04 kJ mol⁻¹ and 168.94 J mol⁻¹ K⁻¹, respectively, which indicated that the electrostatic interactions played a major role in PTA-BSA association. The presence of BSA consistently reduced the inhibitory ability of PTA on AChE, with the relative activity being increased from 46.98 to 61.71% for the concentration range of BSA between 0 and 4.0 g L⁻¹.     

Effect of fenvalerate on the reproduction and fitness costs of the brown planthopper, Nilaparvata lugens and its resistance mechanism

The brown planthopper (BPH), Nilaparvata lugens Stål, is a primary insect pest of cultivated rice, and its effective control is essential for crop production. However, in recent years, outbreaks of the brown planthopper have occurred more frequently in China. In order to determine the causes and mechanisms of insecticide-induced BPH resurgence and perform population management, we conducted the following studies. By the topical application method, our results showed that, fenvalerate acted as stimulus of fecundity from 3.50 × 10⁻³ to 2.02 × 10⁻² μg/female in the BPH. Apart from 7.00 × 10⁻³ μg/female, the number of hatched nymphs was increased gradually with an increase in application dose from 3.50 × 10⁻³ to 1.74 × 10⁻² μg/female. After continuous selection with fenvalerate for 11 generations by the rice-stem dipping method, a resistant strain was achieved with medium resistance to fenvalerate (RR 39.22). Life table study indicated that the resistant strain (G4 and G8) showed reproductive advantages, including increased female ratio, copulation rate and fecundity. But the hatchability of resistant strain was lower. The survival rate and emergence rate were significantly lower in G4 and G8 resistant strain. Resistant strains in G4 and G8 showed a fitness advantage (1.04 and 1.11), and the number of offspring in G8 generation was higher than that in G4 generation. The significant difference detected between resistant insects (G4, G5, G8 and G9) and S-strain contains not only the effect of resistant selection but also the effect of continuous rearing itself. Hence it was concluded that the BPH had the potential to develop high resistance against fenvalerate and the induction of the nymphs by sublethal doses of fenvalerate was of importance in the BPH population management, particularly in the predicting. Further studies demonstrated that triphenyl phosphate (TPP) and diethyl maleate (DEM) had no synergism on fenvalerate. However, piperonyl butoxide (PBO) displayed significant synergism in susceptible strain (1.97) and resistant strain (2.73). We concluded that esterase and glutathione S-transferase play little role in fenvalerate detoxification. The increase of the P450-monooxygenases detoxification is an important mechanism for fenvalerate resistance. Because their resistant populations had a fitness advantage, we should pay close attention to the occurrence of BPH and use other functionally different insecticides to control the BPH.     

抗残杀威和敏感家蝇乙酰胆碱酯酶生化特性研究

对家蝇残杀威抗性(RR)和敏感(SS)品系乙酰胆碱酯酶(acetylcholinesterase,AChE)(RR-和SS-AChE)的生化特性研究后发现,RR- 和SS-AChE存在着明显的差异:1)RR- 和SS-AChE最适反应温度分别为37℃和34℃,但最适pH值均为7.4; 2)AChE催化底物的能力不同,RR-AChE水解碘化硫代乙酰胆碱(ATCh)、碘化硫代丁酰胆碱(BTCh)、碘化硫代丙酰胆碱(PTCh)的活力高于SS-AChE,其相应的最大反应速率(Vmax)比分别是SS-AChE的2.22、1.08和3.41倍; 3)从双分子速率常数(bimolecular constant,Ki)来看,RR-AChE对4种氨基甲酸酯类抑制剂(残杀威、克百威、甲萘威、灭多威)的敏感度分别是SS-AChE的46.77、28.15、66.15和15.00倍,对4种有机磷类抑制剂(马拉氧磷、甲胺磷、氧乐果和氧化三唑磷)的敏感度分别是7.66、12.13、3.81和2.25倍; 4)上述抑制剂与RR-AChE分子相互作用的亲和力常数(Ka)均大于与SS-AChE的值; 5) RR-AChE的磷酰化或氨基甲酰化常数(K2)值都低于SS-AChE的值。 表明RR-AChE的性质已发生变化。     

Influence of three diets on susceptibility of selected insecticides and activities of detoxification esterases of Helicoverpa assulta (Lepidoptera: Noctuidae)

The oriental tobacco worm, Helicoverpa assulta Guenée, is one of the most destructive pests of tobacco and peppers in China. We determined the susceptibility of H. assulta reared on an artificial diet, chili pepper and tobacco to four insecticides (fenvalerate, phoxim, methomyl, indoxacarb) under laboratory conditions associated with the activities of acetylcholinesterase (AChE), carboxylesterase (CarE) and glutathione S-transferase (GST) in its larvae. H. assulta larvae that were fed with chili pepper were more susceptible to fenvalerate, indoxacarb, and phoxim than those that were fed with tobacco and the artificial diet, but not to methomyl. The larvae that were fed with chili pepper were 3.65-, 2.49-, 1.92- and 2.44-fold more susceptible to fenvalerate, phoxim, methomyl, and indoxacarb than those fed with tobacco, respectively. The AChE activities of H. assulta larvae that were fed with chili pepper and tobacco were 2.12 and 1.07 μmol mg⁻¹ 15 min⁻¹, respectively, almost 2-fold difference. The CarE activity of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 4.12, 7.40 and 7.12 μmol mg⁻¹ 30 min⁻¹, respectively. Similarly, the GST activities of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 52.02, 79.37 and 80.02 μmol mg⁻¹ min⁻¹, respectively. H. assulta larvae that were fed with chili pepper were more resistance to the tested insecticides. The low activities of AChE and the high activities of CarE and GST lead to H. assulta become more susceptible to the tested insecticides.     

Determination of carbendazim with multiwalled carbon nanotubes-polymeric methyl red film modified electrode

Multiwalled carbon nanotubes-polymeric methyl red film modified electrode (MWNT-PMRE) was made. The electrochemical behavior of carbendazim on modified electrode was studied with Cyclic Voltammetry, Linear Sweep Voltammetry, Stable Polarization Method and Chronocoulometry. The results indicated that the electrical oxidation of carbendazim on MWNT-PMRE in H₂SO₄ supporting electrolyte with concentration of 0.6 mol/L was irreversible and was mainly controlled by diffusion. Some parameters of the electrochemical process were evaluated. The impacts of experiment conditions on the electrochemical behavior of carbendazim were studied. A good linearity relationship between peak current and concentration of carbendazim in the range of 2.0 × 10⁻⁷-1.0 × 10⁻⁵ mol/L was found, of which the equation was I_p(A) = −1.149 × 10⁻⁵ − 2.301c (mol/L), the correlative coefficient R = −0.9953 and detection limit was 9.0 × 10⁻⁹ mol/L. The recovery was between 90.3% and 94.7%.     

Effects of the organophosphate pesticide Folidol 600® on the freshwater fish, Nile Tilapia (Oreochromis niloticus)

Nile Tilapia (Oreochromis niloticus) juveniles were exposed to different concentrations of Folidol 600® in static toxicity tests. The 24, 48, 72 and 96 h LC₅₀ values of Folidol 600® to O. niloticus were 17.82, 8.91, 4.00 and 2.70 mg L⁻¹, respectively. The values of hematological parameters increased, and inhibition of cholinesterases activity (AChE, BChE and PChE) in plasma of fish exposed to the higher concentrations of pesticide reached 94%. Furthermore, the exposure of Tilapia to Folidol 600® caused an increase of 4%, 20% and 38.4% in oxygen consumption at 0.1, 0.5 and 1.0 mg L⁻¹, respectively. However, exposure to 2.5, 5.0 and 10 mg L⁻¹ caused a decrease of 33.6%, 35.2% and 42.4% in oxygen consumption relative to the control. The ammonium excretion of fish exposed to 0.0, 0.1, 0.5, 1.0, 2.5, 5.0 and 10.0 mg Folidol 600®/L was 0.12, 0.18, 0.30, 0.33, 0.37, 0.36 and 0.33 μg/g/min, i.e., 50%, 150%, 175%, 208%, 200% and 175% increase, respectively, relative to the control.     

Oxygen consumption and ammonia excretion of the freshwater crab Trichodactylus borellianus exposed to chlorpyrifos and endosulfan insecticides

The effects of lethal and sublethal concentrations of chlorpyrifos and endosulfan on oxygen consumption and ammonia excretion rate of the crab Trichodactylus borellianus were evaluated. Oxygen consumption and energy expenditure had significant effect in relation to exposure times. Regarding endosulfan, a significant difference in consumption among times of exposure was registered in 625 μg L⁻¹. Moreover, at the highest concentration, energy expenditure rate was observed stabilized during 1-3 h. A significant increase in ammonia excretion was evidenced in 150 and 300 μg L⁻¹ of chlorpyrifos. The O:N ratio showed a decrease in chlorpyrifos and in 2500 μg L⁻¹ of endosulfan. This indicated a shift towards protein primary metabolism. An increment in the O:N ratio was observed in the lower endosulfan solutions. The relation oxygen:nitrogen showed a shift towards lipid and carbohydrate primary metabolism. This work indicated the complexity of the metabolism in the freshwater crab affected by xenobiotic elements.     

药剂对小菜蛾抗性及敏感品系乙酰胆碱酯酶抑制作用比较

采用浸叶法测定了云南通海、元谋和澜沧的小菜蛾plutella xylostella田间种群对常用杀虫剂的抗药性。结果表明,云南上述地区小菜蛾田间种群对各类杀虫剂均产生了不同程度的抗性。对有机磷类药剂的抗药性为1.74~31.1倍;对菊酯类药剂的抗药性为7.41~764倍;对阿维菌素类药剂则产生了 5.60~4.06×104倍的抗性。通过离体和活体试验测定了药剂对小菜蛾头部乙酰胆碱酯酶(AChE)的抑制作用。敌敌畏和灭多威对通海抗性品系AChE离体和活体内的抑制中浓度(I50)分别是敏感品系的209、26.5倍和2.21、2.16倍;敌敌畏对通海小菜蛾种群的离体和活体内抑制中时间(IT50)小于敏感品系,分别是敏感品系的0.32和0.17倍;而灭多威对通海小菜蛾种群的离体和活体内抑制中时间(IT50)则大于敏感品系,分别是敏感品系的1.37和1.74倍。     

Effects of insecticides on plant-growth-promoting activities of phosphate solubilizing rhizobacterium Klebsiella sp. strain PS19

In this study, four technical grade insecticides, fipronil, pyriproxyfen, imidacloprid and thiamethoxam were applied at the recommended and the higher doses to investigate their effects on plant growth-promoting activities of phosphate-solubilizing Klebsiella sp. strain PS19, isolated from mustard rhizosphere. All tested insecticides displayed a concentration-dependent inhibition in plant growth promoting traits, like, inorganic phosphate solubilization, biosynthesis of phytohormones and siderophores, of rhizobacterial strain PS19. For example, the phosphate-solubilizing activity of Klebsiella sp. PS 19 was reduced maximally by 95%, at 3900 μg l⁻¹ pyriproxyfen over control. At the recommended rate, the magnitude of toxicity of insecticides to plant growth promoting traits was less severe compared to the higher doses. The sequence of insecticide-toxicity expressed as percent decrease, determined at highest dose rate of each insecticide, over control was: pyriproxyfen (95) = imidacloprid (95) > thiamethoxam (94) > fipronil (85), for phosphate-solubilizing activity while for salicylic acid (SA) it was: thiamethoxam > pyriproxyfen = imidacloprid > fipronil. The impact of the highest dose rate of insecticides on 2,3-dihydroxybenzoic acid (DHBA) was almost equal to those observed for SA. Thiamethoxam decreased the indole acetic acid (IAA) synthesis maximally by 86% whereas fipronil had least toxicity and reduced it by 67% relative to the control. Among the experimental insecticides, pyriproxyfen at 3900 μg l⁻¹ in general, had the greatest toxic effects for plant growth promoting activities of the test strain. The study inferred that insecticides affect the plant beneficial activities of rhizobacteria adversely. These findings are likely to add a new insight into the pest management practices.     

Binding of chlorpyrifos and cypermethrin to blood proteins

The features of two insecticides (chlorpyrifos and cypermethrin) binding to two blood proteins, bovine serum albumin (BSA), and bovine hemoglobin (BHb), were investigated via the fluorescence method. The results revealed that both insecticides caused the fluorescence quenching of BSA and the fluorescence enhancement of BHb. A new parameter (F_E), i.e., the fluorescence intensity when adequate insecticide was added, was introduced to obtain the association constant (K_A) and the number of binding sites (n). K_A and n of chlorpyrifos and cypermethrin binding to BSA were 2.99 × 10⁵ and 5.22 × 10⁵ L mol⁻¹, 1.25 and 0.78, respectively. K_A and n of chlorpyrifos and cypermethrin binding to BHb were 2.94 × 10⁴ and 2.48 × 10⁴ L mol⁻¹, 1.75 and 2.19, respectively. In conclusion, chlorpyrifos and cypermethrin could bind to BSA and BHb, and the binding of both insecticides to BSA was significantly stronger than that of insecticides to BHb. These could affect the distribution, metabolism, and excretion of insecticides.     

Effect of waterborne benomyl on the hematological and antioxidant parameters of the Nile tilapia, Oreochromis niloticus

The present study was conducted to determine the 96 h-LC₅₀ of benomyl to the Nile tilapia, Oreochromis niloticus and to investigate the biochemical or hematological indices of blood and the alterations in the antioxidant enzymes of this fish in response to sublethal concentrations of benomyl. Fish weighing 71.61 ± 12.05 g were used in this study; they were subjected to fasting for 4 weeks before treatment. An aqueous solution of benomyl (0, 0.5, 1, 2, 4, 8, and 16 mg L⁻¹) was administered for 96 h to determine the LC₅₀. The 96 h-LC₅₀ value of benomyl was 4.39 (3.23-5.60) mg L⁻¹ in the present study. For 5 weeks, the aqueous solution of benomyl (0, 100, 200, and 400 μg L⁻¹) was administered to investigate its effect on the hematological parameters and antioxidant enzymes. The predominant hematological findings in fish exposed to benomyl were as follows: no significant change in the Hb (g dL⁻¹) level, MCV (μm³), MCH (pg) and MCHC (%) as compared to the control. Benomyl exposure led to greater increases in the GPT, GOT (Karmen-unit), LDH (Wroblewski unit), total cholesterol, Fe, and Ca (mg dL⁻¹) values, whereas the levels of ALP (KA unit), total protein, triglyceride, albumin, and Mg (mg dL⁻¹) did not increase. Benomyl increased the in vivo HSI (%), GST (nmol min⁻¹ mg protein⁻¹), and SOD (U mg protein⁻¹) values in the fish livers in the test group, unlike those in the control group for 5 weeks. At concentrations higher than 100 μg L⁻¹, benomyl affected the GST and SOD levels of Nile tilapia in a dose- and time-dependent manner. The present findings suggest that the in vivo hepatotoxicity associated with benomyl may, in part, result from the hematological index, and antioxidants may provide limited protection against benomyl toxicity.     

Inhibition of Na-K-ATPase in different tissues of freshwater fish Channa punctatus (Bloch) exposed to monocrotophos

Freshwater fish, Channa punctatus, commonly known as the snakehead fish, was exposed to two sublethal concentrations (0.96 and 1.86 mg/L) (selected on the basis of 1/20 and 1/10 of 96 h LC₅₀ value) of monocrotophos for two exposure periods (15 and 60 days). Effects of monocrotophos on Na⁺, K⁺-ATPase in liver, kidney, muscle, intestine, brain, heart and gills were determined. Results indicate that Na⁺, K⁺-ATPase activity in tissues decreased as concentration of monocrotophos and exposure period increased. Monocrotophos induced significant inhibitory effects on the Na⁺, K⁺-ATPase activity of C. punctatus, ranging from gills (70%) > Kidney (63%) > brain (57%) > intestine (52%) > liver (50%) > muscle (47%) > heart (44%) inhibition at a sublethal concentration of 0.96 mg/L. Significant inhibition was detected in Na⁺, K⁺-ATPase activity, ranging from gills (90%) > heart (78%) > kidney (78%) > muscle (74%) > intestine (71%) > brain (67%) > liver (63%) at sublethal concentration of 1.86 mg/L. After subacute exposure (15 days) only gills and brain showed significant inhibition after higher concentration (1.86 mg/L). However, it is evident that exposure duration is more important than dose in the inhibition of the activity of enzyme. At lower concentration initial stimulation of the activity of Na⁺, K⁺-ATPase activity was also noticed. It is suggested that the inhibition of the ATPase by monocrotophos blocked the active transport system of the gill epithelial as well as chloride cells, glomerular and epithelial cells of the tubules and thus altered the osmoregulatory mechanism of the fish. In fact, the impairment of the activity of enzymes which carry out key physiological roles could cause alterations of the physiology of the whole organism.     

Selectivity, acetylcholinesterase inhibition kinetics, and quantitative structure-activity relationships of a series of N-(2-oxido-1,3,2-benzodioxa-phosphol-2-yl) amino acid ethyl or diethyl esters

The inhibitory effects of a recently introduced series of the titled compounds on insect and mammalian acetylcholinesterase (AChE) activity were examined, where the median inhibition concentration (I₅₀) and the inhibition kinetic parameters, bimolecular inhibition rate constant (k_i), affinity constant (K_a), and phosphorylation rate constant (k_p), were determined for each compound. Results indicated that all examined dioxaphospholenes had less inhibitory effects on mammalian AChE than fenitrothion, a commercial pesticide with moderate mammalian toxicity. The highest selectivity was obtained with compounds containing glutamic and leucine moieties (2.70 and 2.18, respectively) while selectivity of fenitrothion was 0.93. The low inhibitory effects of the examined dioxaphospholenes on mammalian AChE were attributed to their low phosphorylation rates (k_p < 2.2 min⁻¹) compared to that of fenitrothion (k_p = 4.84 min⁻¹). QSAR equations indicated that the inhibition process is controlled mainly by both the phosphorylation rate (direct effect) and the affinity of compounds toward the enzyme (inverse effect). Although the compounds’ hydrophobicity had no effects on the inhibition process, it affects the compounds’ toxicity since it affects the ability of compounds to penetrate insects to reach the enzyme active site.     

Inhibitory effects of validamycin compounds on the termites trehalase

Trehalase, with the target to control insects, nematodes and fungi, is of increasing interest and has been investigated extensively in recent years. Validamycin compounds, as competitive trehalase inhibitors and lead compounds with broad applications have attracted substantial attention as well. In this study, the characterizations of termites trehalase were investigated and the inhibitory effects of validamycin compounds on the termites trehalase were studied as well. Results showed that the termites trehalase is presumably belonging to the acid trehalase with optimal pH of 3.3 and optimal temperature of 37 °C. It was investigated that the concentrations of validoxylamine A (VAA), validoxylamine B (VBB), validamycin A (VA) and validamycin B (VB) required for 50% inhibition IC₅₀ of termites trehalase were calculated to be 14.73 mg l⁻¹, 20.80 mg l⁻¹, 3.17 × 10³ mg l⁻¹and 2.24 × 10³ mg l⁻¹, respectively. The inhibition kinetic constant K_i values for the above validamycin compounds were 3.2 × 10⁻⁶ mol l⁻¹, 1.03 × 10⁻⁵ mol l⁻¹_, 4.02 × 10⁻⁴ mol l⁻¹and 2.69 × 10⁻⁴ mol l⁻¹, respectively. Validoxylamine A appeared to be the most potential termites trehalase inhibitor among the four compounds.     

Toxic effects of acephate on Paramecium caudatum with special emphasis on morphology, behaviour, and generation time

The continuous increase in the number of new chemicals as well as the discharges of solid and liquid wastes triggered the need for simple and inexpensive bioassays for routine testing. In recent years, there has been increasing development of methods (particularly rapid tests) for testing environmental samples. This paper describes the quick toxic evaluation of an organophosphorus insecticide, acephate (O,S-dimethyl acetylphosphoramidothioate) on Paramecium caudatum for acute and sub-acute toxicity studies with reference to morphology, behaviour, and its generation time. The lethal concentrations for 10 min and 2 h were determined by probit method, as 500 mg L⁻¹ and 300 mg L⁻¹, respectively. Higher concentrations of 10 min exposure caused cell lysis with disintegration of cell membrane and precipitation of protoplasm. Combination of conventional light microscopy and computerized video tracking systems were used to study the locomotor behaviour of paramecia. The test organism was under stress and exhibited an initial increase and subsequent decrease in the swimming speed when exposed to 1/4, 1/2, 3/4, and LC₅₀ concentrations for 10 min (125, 250, 375, and 500 mg L⁻¹, respectively). Similar changes were also noticed when paramecia were exposed to LC₅₀ for 2 h. In a separate set of experiments, the number of generations and generation time in 24 h was evaluated with respect to the different sub-lethal concentrations (30, 60, 120, and 240 mg L⁻¹). The number of generations decreased and generation time extended significantly in a concentration dependent manner. The results indicate that the Paramecium toxicity assay could be used as a complimentary system to rapidly elucidate the cytotoxic potential of insecticides. The major advantages associated with these tests are: they are inexpensive, simple, user-friendly, space saving, and seem to be attractive alternatives to conventional bioassays.