Alterations of offspring heart muscle electrical activity transferred by rat male genitors chronically treated with lindane (γ-hexachlorocyclohexane) trace concentrations

Using intracellular microelectrodes, we studied transmembrane resting and action potentials (AP) of left ventricle papillary muscles isolated from the heart of adult lindane-treated (TMG) and untreated (UMG) male genitor rat offspring, obtained by mating untreated female with males chronically treated and untreated with lindane (2 ppb) trace concentrations through beverage. The AP magnitude and duration (APD) were similar in both groups and their response to low temperature (22 °C) unchanged. Lowering the external Ca²⁺ concentration from 2.5 to 0.625 mM prolonged APD in the TMG group but not in the UMG group. In the TMG group, (i) cumulative addition of Sr²⁺ (1 mM) to the physiological solution prolonged APD; (ii) apamin (4 μM) and charybdotoxin (4 μM) prolonged the APD. In conclusion, our data revealed that an altered sensitivity of the Ca²⁺-induced Ca²⁺ inactivation of L-type Ca²⁺ channels and of Ca²⁺-activated K⁺ channels to Ca²⁺ has been transferred to TMG offspring.     

Sublethal effects of larval exposure to indoxacarb on reproductive activities of the diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Sublethal effects of larval exposure to indoxacarb on reproductive activities of the diamondback moth, Plutella xylostella (L.) were studied. The third instar male and female larvae of the diamondback moth were fed with collard leaves dipped with a lethal dose of 20% mortality of indoxacarb in order to determine its impact on reproductive activities of adult survivors. Females from indoxacarb treatment had similar peak calling percentage (100%) and mean calling hours (5.83 ± 0.47 h) compared to control females (92% of peak calling, 5.05 ± 0.55 h of mean calling hours) at the first scotophase, but had slightly lower peak calling percentages and shorter calling hours at the subsequent scotophases. The sex pheromone blend or single component from the blend for P. xylostella elicited similar electroantennography responses compared with those from control males and indoxacarb-treated male survivors. Indoxacarb did not alter the number of eggs produced per female and female longevity. However, larval treatment of males with indoxacarb showed a decreased copulation success rate, and this effect was not observed in identically treated female survivors.     

Behavioral hormoligosis in oviposition preference of Bemisia tabaci on cotton

The study on behavioral hormoligosis in oviposition preference in Bemisia tabaci (Genn.) on cotton was conducted, at Entomological Research Farm, Punjab Agricultural University (PAU), Punjab, India, during 2001 crop season and repeated in the same season. Multiple-choice test was followed for conducting the experiment. Quinalphos (250, 375, and 500), carbaryl (625, 938, and 1250), acephate (750, 1125, and 1500), endosulfan (438, 656, and 875), and fenvalerate (25, 38, and 50 g ai/ha) were repeatedly sprayed on potted plants of American cotton (var. LH-1556). The impact of these insecticides was evaluated in term of oviposition preference by B. tabaci to treated plants. Also, it investigated changes in biochemical components of treated cotton leaves and the correlation with oviposition preference. The results revealed that, fenvalerate treated plants were more preferred by whitefly for oviposition. Maximum number of eggs was observed on fenvalerate treated plants, 38, 50, and 25 g/ha (39.3, 37.3, and 36.1 eggs/leaf, respectively) followed by acephate 1500 g/ha (26.9 eggs/ leaf) compared with untreated control (14.1 eggs/leaf). Almost similar trend of results was observed in the repeated experiment. The results obtained from biochemical studies revealed that all the insecticidal treatments caused reduction in total sugars compared with untreated control except fenvalerate and low dose of quinalphos. All insecticides caused increase in total free amino acids and brought significant changes in total phenols and pH value of treated plants. These results have confirmed the behavioral hormoligosis in oviposition preference that induced by fenvalerate and acephate in B. tabaci, which may be one of the causes behind its resurgence on plants repeatedly treated with these insecticides.     

Ultra-structural changes in the integument induced by the use of three of six forms of allylisothiocyanate tested against Plutella xylostella and Pieris rapae larvae

The insecticidal activity of four forms of Hong Jing (HJ) allylisothiocyanate (AITC), AITC + cypermethrin (HJ_A, HJ_B, and HJ_C) with ratio of (1:1, 4:1, and 2:1), pure AITC (HJ_D), and two forms of Hong Du (HD) AITC, AITC + chlorpyrifos (HD_A and HD_B) with ratio of (2:1 and 2:1), respectively, were studied on the major cruciferous insect larvae Plutella xylostella (L.) and Pieris rapae (L.) by combining both spraying and dipping methods. The P. rapae was more susceptible than P. xylostella larvae. The LC₅₀ values 72 h after treatment of AITC forms (HJ_B, HJ_A, HJ_C, HJ_D, HD_B, and HD_A) on the P. rapae were; 0.07, 0.08, 0.16, 0.83, 0.26, 1.08 gL⁻¹, and 0.69, 0.26, 5.45, 0.93, 3.01, 5.98 gL⁻¹ on the P. xylostella, respectively. The toxicity of some of the AITC forms was very close to or better than that of the commercial contact insecticides such as chlorpyrifos (LC₅₀ = 0.03 and 0.04 gL⁻¹ on P. rapae and P. xylostella, respectively), and cypermethrin (0.65 and 0.78 gL⁻¹, respectively, against P. rapae and P. xylostella). The ultrastructural studies on the integument of the third larval instar of P. xylostella treated by sub-lethal concentration (LC₂₀) of HJ_B, HJ_D, and HD_B were carried out by using transmission electron microscope. The more pronounced alterations in the hypodermis and mitochondria cells. They exhibited changes in all treated samples. The hypodermis was almost completely destroyed, and the mitochondria exhibited morphological alterations, represented by enlargement, matrix rarefaction and vacuolization of the mitochondria matrix, quantity of cristae reduced, and density electron matrix lessened. These AITC forms have potential as contact insecticides, and the ultra structural observations confirm the insecticidal efficiency of different AITC forms on P. rapae and P. xylostella.     

Night observations on the cotton leafworm,Spodoptera littoralis: Reliability of pheromone traps for population assessment and efficacy of widely separated pheromone dispensers for mating disruption

Night observations on the cotton leafworm,Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae), were conducted in pheromone-treated and untreated plots in which males, females, mating pairs on plants, and males caught in traps were collected. Results indicated that emergence of a new population was rapid (8-9 days) and that the number of virgin females and of mating pairs on plants reached a peak 5–6 days prior to that of males captured in pheromone traps. Males responded to pheromone traps most actively when the actual virgin female popution had already declined considerably. Dispensers adsorbed with(Z,E)-9,11-tetradecadienyl acetate and located 25 m apart, effectively reduced male captures in traps. However, the number of native mating pairs observed in the treated field during the night was not reduced. Few releasers at wide spacing, even with high amounts of the major pheromonal component, were ineffective in causing mating disruption ofSpodoptera littoralis.     

Sensitivity of the mitotic cells of barley (Hordeum vulgare L.) to insecticides on various stages of cell cycle

In this study, the genotoxic effects of insecticides in different phases of cell cycle were investigated in the mitotic cells of barley (Hordeum vulgare L.). The seeds of H. vulgare L. Var. Karan 4 were treated with different concentrations (0.05%, 0.1% and 0.5%) of insecticides alphamethrin (AM) and monocrotophos (MP) for 6 h after synchronization of cells in G₁, S and G₂ phase of cell cycle with the help of various presoaking durations (7 h, 17 h and 27 h.). Positive and negative control was run parallel in the form of ethyl methane sulphonate (EMS) and distilled water, respectively. The data indicate that higher dose of alphamethrin and monocrotophos produce toxicity, chromosomal aberrations and mitotic aberrations in Hordeum vulgare L. The present study indicates the genotoxic potential of the insecticides AM and MP and it also showed that S-phase of cell cycle was more sensitive compared to the G₁ and G₂ phases.     

Effects of different treatment methods of the fungicide jinggangmycin on reproduction and vitellogenin gene (Nlvg) expression in the brown planthopper Nilaparvata lugens Stål (Hemiptera:Delphacidae)

Jinggangmycin is an antibiotic fungicide against the rice sheath blight, Rhizoctonia solani, in China. Previous investigations have shown that jinggangmycin leaf spray stimulated the fecundity of Nilaparvata lugens Stål (Hemiptera:Delphacidae), but the effects of different application methods and concentrations of jinggangmycin on the reproduction of N. lugens have not been investigated. Here, we investigated three treatment methods (foliar and stem sprays and topical treatment) and four concentrations (100, 200, 400 and 800 ppm) of jinggangmycin on the reproduction of adult female N. lugens. The results showed that leaf spray significantly stimulated the fecundity of N. lugens and increased the vitellin content in female ovaries and the gene expression level of vitellogenin (Nlvg), but there was no significant effect on reproduction found for stem spray. In four concentrations of jinggangmycin, leaf sprays at concentrations of 100, 200 and 400 ppm and topical treatments of 100 and 200 ppm significantly increased the number of eggs laid, the vitellin content and the gene expression level of Nlvg compared to the control. Therefore, we suggest that stem spray at high concentration should be applied (if possible) when jinggangmycin is used against rice sheath blight to facilitate harmonious control of rice pests.     

Effect of temperature on toxicity of pyrethroids and endosulfan, activity of mitochondrial Na-K-ATPase and Ca-Mg-ATPase in Chilo suppressalis (Walker) (Lepidoptera: Pyralidae)

The toxicity of deltamethrin, bifenthrin, and endosulfan to the fourth-instar larvae of rice stem borer, Chilo suppressalis (Walker), was measured at 17, 27, and 37 °C. The three insecticides all displayed a positive temperature coefficient between 17 and 37 °C. The temperature coefficients of deltamethrin, bifenthrin, and endosulfan were 5.59, 1.68, and 2.85, respectively. However, temperature coefficients of deltamethrin and bifenthrin between 17 and 27 °C and between 27 and 37 °C varied. The inhibition of the above three insecticides to mitochondrial Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase from the fourth-instar larvae of rice stem borer was determined at 17, 27, and 37 °C. The inhibition of deltamethrin to the specific activities of Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase showed a negative temperature coefficient, but endosulfan exhibited a positive temperature coefficient. Inhibition of bifenthrin exhibited the contrary temperature coefficients between Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase and a negative temperature coefficient for the former and a positive temperature coefficient for the later.     

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.     

Effects of Artemisia annua L. (Asteracea) on nutritional physiology and enzyme activities of elm leaf beetle, Xanthogaleruca luteola Mull. (Coleoptera: Chrysomellidae)

Methanolic extract of Artemisia annua L., a weed collected around paddy fields in north of Iran, was investigated for its toxic effects on: feeding, growth, fecundity, fertility including the biochemical characteristics of elm leaf beetle Xanthogaleruca luteola Mull. The toxicity tests indicate that insecticidal effect was 2.4-fold higher in 3rd instar larvae than in adults. The LC₅₀ was 48% and 43.77% of a methanolic leaf extract at 24 and 48 h, respectively. The feeding deterrence was highest at 10% concentration and lowest at 0.625% for 3rd instar larvae and adults during the first 24 h and decreased thereafter. Larval duration significantly increased at higher concentrations. Larval treatment with 10% methanolic extract resulted in deformed adults (adultoids) with drastic changes in gross morphology of female reproductive system. Food digestion and assimilation significantly decreased in adults. Larval duration and adult fecundity and fertility were also significantly reduced. Twenty-four hours after treating 3rd instar larva with the extract the levels of glucose, protein, urea, uric acid, α-amylase, alkaline phosphatase, alanine amino transferase and aspartate amino transferase significantly changed. However, at 48 h the extract lost its potency. The extract did not effect the level of cholesterol in treated larva. These results indicate that Artemisia annua L. extract contains inhibitors of key metabolic pathways that may be useful in future control of the elm leaf beetle.     

Binding of three Cry1A toxins in resistant and susceptible strains of cotton bollworm (Helicoverpa armigera)

Evolution of resistance by pests is the greatest threat to the continuous success of theBacillus thuringiensis (Bt) toxins used in conventional sprays or in transgenic plants. The most common mechanism of insect resistance to Bt is reduced binding of toxins to target sites in the brush border membrane of the larval mid-gut. In this paper, binding experiments were performed with three ¹²⁵I-Cry1A toxins and the brush border membrane vesicles from Cry1Ac resistant or susceptible strains of Helicoverpa armigera. The homologous competition test showed that there was no significant difference in Cry1Ac-binding affinity, but the concentration of Cry1Ac-binding sites dramatically decreased in the resistant strain (R_t decreased from 5.87 ± 1.40 to 2.23 ± 0.80). The heterologous competition test showed that there were three Cry1Ac-binding sites in the susceptible strain. Among them, site 1 bound with all three Cry1A toxins, site 2 bound with both Cry1Ab and Cry1Ac, and site 3 only bound with Cry1Ac. In the Cry1Ac resistant strain, the binding capability of site 1 with Cry1Ab decreased and site 2 did not bind with Cry1Ac. It is suggested that the absence of one binding site is responsible for H. armigera resistance to Cry1Ac. This result also showed that the resistance fitted the “mode 1” pattern of Bt resistance described previously.     

Effect of <Emphasis Type="Italic">Metarhizium guizhouense</Emphasis> infection on mating competition and mate choice of <Emphasis Type="Italic">Bactrocera latifrons</Emphasis> (Diptera: Tephritidae)

Metarhizium guizhouense PSUM02 treated males of Bactrocera latifrons were investigated for the mating competition among males and mating choice by female flies to develop an auto-dissemination for the control of B. latifrons. In the present study, on day 1–4 of experiment, M. guizhouense–treated male flies were equally competitive with the normal male flies as we did not observe any differences in mating by treated and normal male flies of B. latifrons. Further, mating competitiveness were found low in treated adult male B. latifrons than normal male B. latifrons from 5th days of treatment until death. Kaplan-Meier survival analysis of treated male flies gave average survival times (AST) of 4.3?±?0.1 days, while the healthy female and male flies in the same cage showed AST of 9.3?±?0.3 and 8.3?±?0.4 days, respectively. The AST of untreated flies in control experiment ranged from 14.2–14.5 days. In mating preference experiment, M. guizhouense–treated male flies were chosen by virgin female than gravid female flies for mating. The treated male flies caused mortality in both virgin and gravid female flies in the same cage with AST of 4.4?±?0.1, 5.6?±?0.1 and 7.4?±?0.2 days, respectively, while untreated flies showed AST ranged from 13.9–14.3 days in control. The treated male flies could transmit the fungus infection to both untreated female and male flies as well as in virgin and gravid female flies by mating and contact. Our experiments showed the potentiality of M. guizhouense PSUM02 in management of B. latifrons by auto-dissemination with treated male flies, which transmit the fungus to a healthy population to reduce insect pest infestations.     

Azinphos-methyl and carbaryl resistance in adults of the codling moth (Cydia pomonella (L.), Lepidoptera: Tortricidae) from Northeastern Spain

The resistance of Cydia pomonella (L.) to organophosphates is widespread throughout the pome fruit growing areas. The lethal effects of two insecticides inhibitors of the acetylcholine esterase, azinphos-methyl and carbaryl, were evaluated in adults of five and four field populations of the codling moth, respectively. The lethal concentrations (LC₅₀ and LC₉₀) of these insecticides were determined in a susceptible strain from Spain (S_Spain). Topical bioassays using the approximate LC₉₀ values (3000 mg (a.i.)/L of carbaryl and 2000 mg (a.i.)/L of azinphos-methyl) that were obtained in S_Spain were tested as diagnostic concentrations. The enzymatic activities of mixed-function oxidases (MFO), glutathione S-transferases (GST) and esterases (EST) were measured to investigate their potential role in the detoxification of these insecticides.Carbaryl and azinphos-methyl caused ?53% and ?39% corrected mortality, respectively, in field populations, although the diagnostic concentrations applied were twofold and fourfold higher than the maximum concentration registered in Spain, respectively. The activities of MFO and GST were 7.3- to 16.1-fold higher and 2.5- to 3.7-fold higher in all the field populations compared to those in S_Spain, respectively.     

Toxicity and neurophysiological effects of selected insecticides on the mole cricket, Scapteriscus vicinus (Orthoptera: Gryllotalpidae)

Mole crickets (Scapteriscus spp.) are severe subterranean pests of turfgrasses, commonly targeted with neurotoxic insecticides. Ideally insecticides used against mole crickets should induce quick knockdown or mortality to minimize damage caused by their tunneling. However, neurophysiological effects of insecticides on mole crickets are mostly unknown. The aims of this study were to investigate neurophysiological and toxic effects of several insecticides on tawny mole cricket (Scapteriscus vicinus Scudder) adults and nymphs, and potential synergy between pyrethroid and neonicotinoid insecticides. Bifenthrin, fipronil, and the combination of bifenthrin + imidacloprid provided the fastest median mortality when injected. The combination of bifenthrin + imidacloprid elicited faster toxicity than either active ingredient alone. Imidacloprid, bifenthrin, and bifenthrin + imidacloprid caused immediate knockdown, whereas fipronil immobilized mole crickets within 1-2 h. Acephate, bifenthrin, fipronil, imidacloprid, and bifenthrin + imidacloprid caused significant neuroexcitation. Bifenthrin + imidacloprid resulted in greater increases of spontaneous neural activity than the additive effects of imidacloprid and bifenthrin alone. Excitatory compounds acting at sodium and chloride channels (bifenthrin and fipronil) were the most toxic against S. vicinus. Combining a sodium channel toxin (bifenthrin) and a synaptic toxin (imidacloprid) led to greater than additive neurophysiological and toxic effects, which to our knowledge provides the first documented evidence of synergistic neurological “potentiation”.     

Effect of Basudin, Selecron and the phytoalkaloid Colchicine (pesticides) on biological and molecular parameters of Biomphalaria alexandrina snails

The results showed that survival rates of Biomphalaria alexandrina snails, reproductive potential and hatchability of eggs were evaluated post exposure to Basudin, Selecron and Colchicine. As well, DNA and RNA changes in the cells of ovotestis-digestive gland complex of treated snails were estimated. The current molluscicide Bayluscide was used as a reference compound.The pesticide Selecron proved to be more toxic to B. alexandrina snails than Basudin and Colchicine. Juvenile snails were dead post 3 weeks of exposure to the sublethal concentration LC₀ of either Selecron or Basudin, while 26.75% of snails still alive at Bayluscide treatment. In addition, exposure of adult snails to LC₀ of Selecron for 24 h/week for 4 weeks markedly reduced their reproductive rate (R₀) by 89.9%. Moreover, snails’ eggs failed to hatch post 24 h of exposure to LC₉₀ of either Selecron, Basudin or Bayluscide. Electrophoretic analysis indicated a decrease in the molecular weight of intact DNA in the ovotestis-digestive gland complex of snails treated with 250 ppm of Colchicine and LC₂₅ of Selecron, as it scored 1.2 and 76 bp, respectively, compared to 166.46 bp for control group, while the vice versa was recorded for RNA intensity. It was concluded that the tested pesticides have deleterious effects on snails’ reproductive rate, their eggs and the intensities of DNA and RNA in their ovotestis-digestive gland complex. Therefore, it is expected that reaching of such pesticides to snails’ habitats in water courses during plant pests control could minimize the population density of the snails intermediate hosts of schistosomiasis, hence probably interrupt and reduce the transmission of this parasite.     

Mating,longevity, fertility and fecundity of the cotton leaf-worm,Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae)

The mechanisms adopted to ensure mating success, fertility and fecundity ofS. littoralis were studied. The male moths usually mate only once a night but they mate repeatedly during most of their life span, and practically all their spermatophores are equally efficient for inseminating a female. Although females are also multiple maters, one spermatophore is sufficient for a female to lay fertile eggs throughout her life with no reduction in potential fecundity. While a male may mate 5.3 times, on the average, a female may mate only 2.3 times. As a pronounced decrease in pheromone release occurs after mating, a repeat mating by a female will occur only after most neighboring competitive virgin females have been inseminated. A male may mate therefore with 5.3 different females, all of which will be thoroughly inseminated. The female moth is ready to mate just after emergence and at any time throughout her life. If mating is delayed the preoviposition period is greatly prolonged and eggs are saved to be laid later, after mating, as fertile eggs. Delay in egg laying due to delayed insemination does not result in reduced fecundity as it is compensated for by prolonged longevity. Even if mating is greatly delayed and occurs after egg laying has already begun, the potential reduction in reproduction is not very great, due to the low daily egg-laying rate and the increased longevity of unmated females. It appears unreasonable to expect any tendency toward reduction of egg production with a decrease in the ratio of adult males in a population; for this purpose the control of males must be essentially complete. Therefore, a behavioral control program by mass trapping ofS. littoralis males is unlikely to be economically feasible.     

Purification and partial characterization of a glutathione S-transferase from the two-spotted spider mite, Tetranychus urticae

Glutathione S-transferases (GSTs) are known to catalyze conjugations by facilitating the nucleophilic attack of the sulfhydryl group of endogenous reduced glutathione on electrophilic centers of a vast range of xenobiotic compounds, including insecticides and acaricides. Elevated levels of GSTs in the two-spotted spider mite, Tetranychus urticae Koch, have recently been associated with resistance to acaricides such as abamectin [Pestic. Biochem. Physiol. 72 (2002) 111]. GSTs from acaricide susceptible and resistant strains of T. urticae were purified by glutathione-agarose affinity chromatography and characterized by their Michaelis-Menten kinetics towards artificial substrates, i.e., 1-chloro-2,4-dinitrobenzene and monochlorobimane. The inhibitory potential of azocyclotin, dicumarol, and plumbagin was low (IC₅₀ values > 100 μM), whereas ethacrynic acid was much more effective, exhibiting an IC₅₀ value of 4.5 μM. GST activity is highest in 2-4-day-old female adults and dropped considerably with progressing age. Furthermore, molecular characteristics were determined for the first time of a GST from T. urticae, such as molecular weight (SDS-PAGE) and N-terminal amino acid sequencing (Edman degradation). Glutathione-agarose affinity purified GST from T. urticae strain WI has a molecular weight of 22.1 kDa. N-terminal amino acid sequencing revealed a homogeneity of ≈50% to insect GSTs closely related to insect class I GSTs (similar to mammalian Delta class GSTs).     

Thiamethoxam is a neonicotinoid precursor converted to clothianidin in insects and plants

Neonicotinoid insecticides are compounds acting agonistically on insect nicotinic acetylcholine receptors (nAChR). They are especially active on hemipteran pest species such as aphids, whiteflies, and planthoppers, but also commercialized to control many coleopteran and some lepidopteran pest species. The most prominent member of this class of insecticides is imidacloprid. All neonicotinoid insecticides bind with high affinity (I₅₀-values around 1 nM) to [³H]imidacloprid binding sites on insect nAChRs. One notable ommission is the neonicotinoid thiamethoxam, showing binding affinities up to 10,000-fold less potent than the others, using housefly head membrane preparations. Electrophysiological whole cell voltage clamp studies using neurons isolated from Heliothis virescens ventral nerve cord showed no response to thiamethoxam when applied at concentrations of 0.3 mM, although the symptomology of poisoning in orally and topically treated noctuid larvae suggested strong neurotoxicity. Other neonicotinoids, such as clothianidin, exhibited high activity as agonists on isolated neurons at concentrations as low as 30 nM. There was no obvious correlation between biological efficacy of thiamethoxam against aphids and lepidopterans and receptor affinity in electrophysiological and binding assays. Pharmacokinetic studies using an LC-MS/MS approach to analyze haemolymph samples taken from lepidopteran larvae revealed that thiamethoxam orally applied to 5th instar Spodoptera frugiperda larvae was rapidly metabolized to clothianidin, an open-chain neonicotinoid. Clothianidin shows high affinity to nAChRs in both binding assays and whole cell voltage clamp studies. When applied to cotton plants, thiamethoxam was also quickly metabolized, with clothianidin being the predominant neonicotinoid in planta briefly after application, as indicated by LC-MS/MS analyses. Interestingly, the N-desmethylated derivative of thiamethoxam, N-desmethyl thiamethoxam, was not significantly produced in either lepidopteran larvae or in cotton plants, although it was often mentioned as a possible metabolite, being nearly as active as imidacloprid. In conclusion, our investigations show that thiamethoxam is likely to be a neonicotinoid precursor for clothianidin.     

The effect of single, binary, and tertiary combination of few plant derived molluscicides alone or in combination with synergist on different enzymes in the nervous tissues of the freshwater snail Lymnaea (Radix) acuminata (Lamark)

The effect of single, binary, and tertiary combination of few plant derived molluscicides alone or in combination with synergist on different enzymes (acetylcholinesterase—AChE, lactic dehydrogenase—LDH, and acid/alkaline phosphatase—ACP/ALP in the nervous tissue of the freshwater snail Lymnaea acuminata were studied. Sublethal in vivo 24 h exposure to 40 and 80% of LC₅₀ of Azadirachta indica oil (AI), oleoresin of Zingiber officinale (OL), Cedrus deodara oil (CD), Allium sativum (AS), and Polianthes tuberosa (PT) bulb powder singly, their binary combination of AI + OL, AS + CD, AS + PT, CD + OL, CD + PT, OL + PT, and tertiary combination of these binary combinations with the synergist piperonyl butoxide (PB) or MGK-264 significantly altered the activity of these enzymes. Tertiary combination with PB or MGK was very effective. Combination of CD + PT + MGK was more effective against AChE whereas, the combination of CD + OL + PB, CD + AS + PB, and CD + PT + PB were more effective against LDH, ACP, and ALP, respectively.