Phytoremediation of cadmium-trichlorfon co-contaminated water by Indian mustard (Brassica juncea): growth and physiological responses.

In this study, the morphological and physiological responses of Brassica juncea to the stresses of Cadmium (Cd) and trichlorfon (TCF), and the phytoremediation potential of B. juncea to Cd and TCF were investigated under hydroponics. Results showed that Cd exhibited strong inhibition on biomass and root morphology of B. juncea as Cd concentration increased. The chlorophyll a fluorescence intensity and chlorophyll content of B. juncea decreased with the increased Cd concentration, whereas the malondialdehyde and soluble protein contents and superoxide dismutase activity increased. TCF with different concentrations showed no significant influence on these morphological and physiological features of B. juncea. The biomass and physiological status of B. juncea were predominantly regulated by Cd level under the co-exposure of Cd and TCF. B. juncea could accumulate Cd in different plant parts, as well as showed efficient TCF degradation performance. A mutual inhibitory removal of Cd and TCF was observed under their co-system. The present study clearly signified the physiological responses and phytoremediation potential of B. juncea toward Cd and TCF, and these results suggest that B. juncea can be used as an effective phytoremediation agent for the Cd-TCF co-contamination in water.

Combined pollution of heavy metals and pesticides in agricultural water systems is a common phenomenon. In previous phytoremediation studies, limited information is available on the co-contamination of heavy metals and pesticides. In this study, we aimed to investigate the concentration-dependent morphological and physiological characteristics of B. juncea under single and co-stress of Cd and trichlorfon (TCF), and the phytoremediation ability of B. juncea to remove Cd and TCF through hydroponic experiment. B. juncea exhibited efficient removal performance of Cd and TCF alone and simultaneous exposure of both pollutants, indicating that B. juncea is an effective phytoremediation agent for the Cd-TCF co-contaminated water.

The Combined Use of Copper Sulfate and Trichlorfon Exerts Stronger Toxicity on the Liver of Zebrafish.

In aquaculture, copper sulphate and trichlorfon are commonly used as disinfectants and insecticide, sometimes in combination. However, improper use can result in biotoxicity and increased ecological risks. The liver plays a crucial role in detoxification, lipid metabolism, nutrient storage, and immune function in fish. Selecting the liver as the main target organ for research helps to gain an in-depth understanding of various aspects of fish physiology, health, and adaptability. In the present study, zebrafish were exposed to Cu (0.5 mg/L) and Tri (0.5 mg/L) alone and in combination for 21 days. The results demonstrate that both Cu and Tri caused hepatocyte structure damage in zebrafish after 21 days of exposure, with the combination showing an even greater toxicity. Additionally, the antioxidant and immune enzyme activities in zebrafish liver were significantly induced on both day 7 and day 21. A transcriptome analysis revealed that Cu and Tri, alone and in combination, impacted various physiological activities differently, including metabolism, growth, and immunity. Overall, Cu and Tri, either individually or in combination, can induce tissue damage by generating oxidative stress in the body, and the longer the exposure duration, the stronger the toxic effects. Moreover, the combined exposure to Cu and Tri exhibits enhanced toxicity. This study provides a theoretical foundation for the combined use of heavy metal disinfectants and other drugs.

Trichlorfon blocks androgen synthesis and metabolism in rat immature Leydig cells.

Trichlorfon is a widely used organophosphorus insecticide. It has been reported that it has reproductive toxicity to animal models. However, whether trichlorfon affects testosterone biosynthesis and metabolism remains unclear. In this study, we explored the effects of trichlorfon on the steroidogenesis and the expression of genes in androgen biosynthetic and metabolic cascades in immature Leydig cells isolated from pubertal male rats. Immature Leydig cells were treated with trichlorfon (0.5-50 µM) for 3 h. Trichlorfon significantly inhibited total androgen output under basal condition at 5 and 50 µM, and under LH- and cAMP-stimulated conditions at 50 µM. Trichlorfon also downregulated the expression of Star, Sod2, and Gpx1 and their proteins at 5 and 50 µM and the expression of Cyp11a1, Hsd3b1, Cyp17a1, and Srd5a1 at 50 µM. Trichlorfon significantly inhibited total androgen output at 50 µM, which was partially reversed by 400 µg/ml vitamin E, which alone had no effects on androgen output. In conclusion, trichlorfon downregulates the expression of steroidogenesis-related genes and antioxidants, which leads to a decrease in androgen production in rat immature Leydig cells.

In vitro and in vivo action of turmeric oil (Curcuma longa L.) against Argulus spp. in goldfish (Carassius auratus).

Background: Argulus is a common and widespread ectoparasite that causes major parasitic diseases and is a virus and bacteria carrier in the ornamental fish trade. Aim: The purpose of this study is to determine what types of phytochemicals are present in the essential oil extracted from turmeric rhizome (Curcuma longa L.) and to assess the efficacy of turmeric oil in eliminating Argulus spp. infestations in goldfish (Carassius auratus). Methods: The chemical composition and quantity of the major substances in essential oils from fresh turmeric rhizome were detected by gas chromatography/mass spectrometry (GC-MS). The antiparasitic effect of turmeric oils on Argulus spp. was tested at 12.5, 25, 50, 100, and 200 ppm and compared to 0.25 ppm Neguvon® (the positive control). The percentage of Argulus spp. killed, the percentage of the mean mortality rate, and the effectiveness of each test were evaluated. Results: By using GC-MS analysis, it was possible to identify the primary phytochemical component of turmeric essential oil as b-turmerone. The results obtained from the in vitro test indicated that there was a correlation between the concentration of turmeric essential oil and the average mortality rate of fish lice. The mean mortality of fish louse exposed to 200 ppm turmeric essential oil was higher than the mean mortality of fish louse exposed to Neguvon® (p < 0.05). In an in vivo study, the effectiveness of 12.5 ppm turmeric essential oil against parasites was 44.44%, 55.46%, and 62.83% at 24, 48, and 72 hours, respectively. Conclusion: In summary, the efficacy of turmeric essential oil against fish louse has been shown both in vitro and in vivo studies.

Effects of Sublethal Concentrations of Insecticides on Survival and Reproduction of Two Bactrocera Species (Diptera: Tephritidae).

The melon fruit fly, Bactrocera cucurbitae (Coquillett) and the pumpkin fruit fly, Bactrocera tau (Walker) (Diptera: Tephritidae), are important invasive pests on Cucurbitaceous hosts. The acute toxicity of five insecticides to females of these two fruit fly species was bio-assayed by membrane method, and effects of their sublethal concentration on the survival, reproduction, and ovary development of females were evaluated in the laboratory. Results showed that based on the LC50 values, both B. cucurbitae and B. tau were the most susceptible to trichlorfon. After treatment with sublethal concentration (LC30) of trichlorfon, abamectin+ß-cypermethrin, spinetoram, and lambda-cyhalothrin, the female survival days of the two Bactrocera species were significantly shortened compared with the control. Moreover, the fecundity of two Bactrocera species was remarkably decreased, after exposure to abamectin+ß-cypermethrin and trichlorfon LC30. However, the sublethal concentration (LC30) of insecticides had no significant influence on the egg hatchability of the fruit flies. Furthermore, after treatment with abamectin+ß-cypermethrin LC30, the ovary length, width, and egg load of B. cucurbitae were significantly lower than that of the control; however, only the ovarian length and egg load of B. tau were significantly decreased on the 16th and 20th day. In conclusion, abamectin+ß-cypermethrin has an excellent insecticidal activity against B. cucurbitae and B. tau.

Effects of low doses Trichlorfon exposure on Rana chensinensis tadpoles.

Trichlorfon is an organophosphate insecticide widely used in aquaculture and agriculture. Little is known about the effects of long-term of low doses trichlorfon exposure on amphibians. In this study, we investigated the effects of low doses trichlorfon on Rana chensinensis tadpoles after exposure to 0.01, 0.1, and 1.0 mg/L trichlorfon for 2 and 4 weeks. Survival, growth, development and mortality were monitored regularly over the course of exposure. The results showed that trichlorfon led to a decrease in tadpole survival. Reductions in growth and disruptions to the development of tadpoles were observed in trichlorfon treatments. Morphological abnormalities of affected tadpoles included axial flexures, skeletal malformations and lateral kinks. Trichlorfon increased the frequency of micronucleus (MN) formation in circulating erythrocytes of tadpoles exposed for 2 weeks to 0.1 and 1.0 mg/L trichlorfon. At all concentrations, an enhanced frequency of MN formation was observed in tadpoles exposed for 4 weeks. Exposure to trichlorfon induced other nuclear abnormalities such as lobed and notched nuclei only in tadpoles exposed to 1.0 mg/L trichlorfon for 4 weeks. In addition, exposure to trichlorfon within the 0.01-1.0 mg/L range increased the genetic damage index in hepatic tissues in all treatments. Apoptosis-associated DNA fragmentation in hepatic tissues occurred in a weak ladder-like pattern. This study presents evidence of low doses trichlorfon effects on amphibians, highlighting the properties of this organophosphate insecticide that jeopardize nontarget species exposed to trichlorfon.

Trichlorfon and spinosad resistance survey and preliminary determination of the resistance mechanism in Pakistani field strains of Bactrocera dorsalis.

The use of insecticides has been a primary tool to manage Bactrocera dorsalis in Pakistan; however, recent reports of field control failures necessitate mapping out the insecticide resistance problem. Therefore, eight field strains from Pakistan, were evaluated for their resistance against trichlorfon and spinosad. Compared with a reference strain, six field strains showed high levels of resistance to trichlorfon, while two field strains expressed intermediate resistance. In case of spinosad, five field strains fell in the susceptible range, whereas, the rest of the strains represented minor resistance. Correlation analysis between LD50 values of trichlorfon and spinosad of all the field strains revealed non-significant association, suggesting the possibility of lack of cross-resistance between both insecticides. Synergism bioassays implementing S,S,S-tributylphosphorotrithioate (DEF) and piperonyl butoxide (PBO) revealed that the LD50 values of trichlorfon in the presence of either DEF or PBO in seven field strains were significantly reduced. However, DEF and PBO had a non-significant effect on synergizing spinosad toxicity. The results revealed resistance to trichlorfon in field strains of B. dorsalis, which might be metabolic-based. Absence or minor resistance to spinosad and lack of cross-resistance to trichlorfon, suggest that spinosad could be a potential candidate for managing B. dorsalis.

In vitro effect of metrifonate on the indices of oxidative stress in Gigantocotyle explanatum.

Helminth infections in general and digenetic trematodes in particular cause a huge economic loss globally to our livestock. Gigantocotyle explanatum is a digenetic amphistome that infects the bile ducts of water buffalo and are highly prevalent in tropical and sub-tropical countries. In the present study, effects of an organophosphate compound, Metrifonate (MF) in three doses, viz., 9.4 × 10-5 M (Dose I), 14.4 × 10-5 M (Dose II), and 19.4 × 10-5 M (Dose III), have been studied in vitro, on the motility and on some enzymatic and non-enzymatic oxidative stress indices in G. explanatum. The worm's motility and their non-enzymatic oxidative stress biomarkers like lipid peroxides measured as thiobarbituric acid-reactive substances (TBARS) and reduced glutathione (GSH) were disrupted significantly in a dose-dependent manner. However, the enzymatic oxidative stress biomarkers like glutathione-S-transferase (GST) and superoxide dismutase (SOD) were affected by MF treatment in a biphasic manner. Exposure to Dose I significantly stimulated the activities of both GST and SOD, whereas exposure to Doses II and III resulted into significant inhibition in a dose-dependent manner. Our findings suggest that MF has potential to be a strong and effective anthelmintic, however, further studies in vitro as well as in vivo are needed to explore further these observations and understand the exact mode of MF action in G. explanatum and other trematodes of veterinary economic importance.

Gut symbiont enhances insecticide resistance in a significant pest, the oriental fruit fly Bactrocera dorsalis (Hendel).

BACKGROUND: Symbiotic bacteria affect insect physiology and ecology. They may also mediate insecticide resistance within their hosts and thereby impact pest and vector control practices. Here, we document a novel mechanism of insecticide resistance in which a gut symbiont of the tephritid pest fruit fly Bactrocera dorsalis enhances resistance to the organophosphate insecticide trichlorphon. RESULTS: We demonstrated that the gut symbiont Citrobacter sp. (CF-BD) plays a key role in the degradation of trichlorphon. Based on a comparative genomics analysis with other Citrobacter species, phosphatase hydrolase genes were identified in CF-BD. These CF-BD genes had higher expression when trichlorphon was present. Bactrocera dorsalis inoculated with isolated CF-BD obtained higher trichlorphon resistance, while antibiotic-treated flies were less resistant confirming the key role of CF-BD in insecticide resistance. CONCLUSIONS: Our findings suggest that symbiont-mediated insecticide resistance can readily develop in B. dorsalis and may represent a more widely relevant insecticide resistance mechanism than previously recognized.

Effects of Hydrogen Peroxide and Metrifonate on Monogenean ligictaluridus floridanus on Catfish ( Ictalurus punctatus , Rafinesque) Gills.

Hydrogen peroxide (H2O2) and metrifonate (Mtf) are common products used in ectoparasite infestations on fish cultures. The therapeutic efficacy of H2O2 and Mtf on a common monogenean parasite, Ligictaluridus floridanus, was evaluated in channel catfish ( Ictalurus punctatus ). In vitro trials were conducted using excised fish gills naturally infected with L. floridanus, which were immersed in H2O2 (150, 300, and 570 mg L(-1)) and Mtf (0.25, 0.5, and 0.75 mg L(-1)) solutions. The efficacy of the treatments was based on the survival time of the parasites, observed microscopically. In addition, an in vivo trial using catfish juveniles, naturally infected with L. floridanus, was also performed. One group received immersion baths of 570 mg L(-1) H2O2 (3%) during 4 min; the Mtf (90%) group received 0.5 mg L(-1) Mtf for 10 min. Treatments were done on days 3, 7, and 11 of the experiment. Results indicate that baths with Mtf do not significantly reduce the mean intensity of the parasite per gill arch, nor do they reduce the in vitro survival time of parasites during treatment; H2O2 baths at 570 mg L(-1) during 4 min were effective (P < 0.05) against adult and juvenile stages of L. floridanus. This study supports the use of H2O2 as an effective antiparasitic agent against I. punctatus .

Metabolic effects of trichlorfon (Masoten®) on the neotropical freshwater fish pacu (Piaractus mesopotamicus).

Fish parasites are among the crucial limiting factors in aquaculture. The organophosphorous pesticide trichlorfon is widely used as an insecticide and against fish parasites worldwide. In this study, the effects of environmental trichlorfon on biochemical and physiological parameters were investigated in Piaractus mesopotamicus (pacu), a widely farmed fish in South America, through sublethal exposure (8 µg L(-1), 10 % of the LC50; 96 h) and recovery. The activity of brain acetylcholinesterase (AChE) was reduced after exposure (15.5 %) and remained decreased during the recovery (21.5 %). In white muscle, AChE activity decreased 31 % only after recovery. Alkaline phosphatase (ALP) and acid phosphatase (ACP) activities of the liver, muscle and plasma were steady during exposure. However, after the recovery period, ALP activity was increased in the liver and muscle and decreased in plasma, while ACP was increased in the liver and decreased in muscle. Intermediary metabolism was also affected by trichlorfon, depicting increase of energetic demand (hypoglycemia, neoglucogenesis and lipid catabolism), which remained even after recovery. These results indicate that P. mesopotamicus is adversely affected by sublethal concentrations of trichlorfon and are useful for assessing the impact as well as the pros and cons of its use in controlling fish parasites in aquaculture.

Metrifonate, like acetylcholine, up-regulates neurotrophic activity of cultured rat astrocytes.

BACKGROUND: Metrifonate is an inhibitor of acetylcholinesterase (AChE). Several studies confirmed its positive effects on cognitive impairment in Alzheimer's disease but it was due to adverse events withdrawn from clinical trials. Based on the importance of astrocytes in physiological and pathological brain activities we investigated the impact of metrifonate and, for comparison, acetylcholine on intrinsic neurotrophic activity in these cells. METHODS: Metabolic activity, intracellular adenosine 5'-triphosphate (ATP) levels and lactate dehydrogenase (LDH) release was measured to examine the impact of metrifonate on viability and integrity of cultured rat cortical astrocytes. The influence of metrifonate, acetylcholine and selective cholinergic ligands on nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) synthesis and secretion was determined by specific two-site enzyme immunoassays. RESULTS: Exposure of cultured astrocytes to metrifonate displayed no toxic effects on cell viability. Metrifonate and acetylcholine potently and transiently elevated NGF and BDNF, but not NT-3, protein levels and secretion with different intensity and time frame of their maximal response. Stimulatory effect on NGF was mimicked by selective nicotinic receptor agonist nicotine and completely blocked by nicotinic antagonist mecamylamine. The impact on BDNF synthesis was mimicked by muscarinic receptor agonist pilocarpine and abolished by selective muscarinic antagonist scopolamine. CONCLUSIONS: Metrifonate up-regulates astrocytic NGF and BDNF synthesis in the same manner as acetylcholine, their effect depends on different cholinergic pathways. These results suggest a trophic role of metrifonate, based on a well-known neurotrophic activity of NGF and BDNF in vivo.

Molecular and immunological responses of the giant freshwater prawn, Macrobrachium rosenbergii, to the organophosphorus insecticide, trichlorfon.

Trichlorfon is an organophosphorus (OP) insecticide that is used as an agriculture pesticide to destroy insects, a human medicine to combat internal parasites, and an ectoparasiticide in the livestock and aquaculture industries, but which has caused aquatic toxicity in the prawn industry. The aim of this study was to investigate the effects of trichlorfon on molecular and enzymatic processes of the immunological response of the giant freshwater prawn, Macrobrachium rosenbergii, at 0, 0.2, and 0.4mgL(-1) with 0, 3, 6, 12, and 24h of exposure. The total hemocyte count (THC), respiratory bursts (RBs), phenoloxidase (PO) activity, and superoxide dismutase (SOD) activity were examined to evaluate immunological responses and oxidative stress. Results showed that THCs of the prawn exposed to trichlorfon at both concentrations (0.2 and 0.4mgL(-1)) had increased after 12 and 24h; SOD and PO activities had significantly increased at 3h, whereas production of RBs had dramatically increased as oxidative stress at each sampling time after exposure to trichlorfon compared to the control. A potential biomarker of OPs, acetylcholinesterase (AChE) revealed a significant decrease after exposure for 6h, and showed a time-dependent tendency. Immune gene expressions, including prophenoloxidase (proPO), the lipopolysaccharide- and ß-1,3-glucan-binding protein (LGBP), peroxinectin (PE), α2-macroglubulin (α2M), transglutaminase (TG), and copper, zinc (Cu,Zn)-SOD, of prawns exposed to trichlorfon at 0, 0.2, and 0.4mgL(-1) for 0, 6, and 24h were further evaluated. Expressions of all of the immune genes significantly decreased when prawns were exposed to 0.4mgL(-1) trichlorfon for 24h, and among them, an increase in SOD expression was seen after exposure to 0.4mgL(-1) for 6h. Prawns exposed to trichlorfon within 24h exhibited the decrease of circulating hemocytes, and also the induction of oxidative stress, which caused subsequent damage to DNA formation of immune genes. From these results, we concluded that immunological responses and immune gene expressions of prawn exposed to trichlorfon at 0.4mgL(-1) for 24h were perturbed, thus causing a deficiency in immunity and subsequent increased susceptibility to pathogen infections.

Characteristics of protein variants in trichlorphon-resistant Bactrocera dorsalis (Diptera; Tephritidae) larvae.

Functional proteins in larvae of Bactrocera dorsalis, a major fruit pest, play a central role in their resistance to organophosphorus insecticides. Changes in proteins in B. dorsalis larvae after trichlorphon treatment may have a role in the resistance response to trichlorphon. We analyzed 14 protein spots of crude proteins from B. dorsalis larvae post-treatment with trichlorphon in two-dimensional gel electrophoresis through mass spectrometry and protein sequencing. We found functional proteins that are responsible for signal transduction (pkaap and dual specificity tyrosine-phosphorylation-regulated kinase), immunity (hemolectin), synthesis and decomposition (twinstar, cathepsin B, RE66325p), oxidative stress metabolism (glutathione S transferase and CG7320), energy metabolism (Act57B), and cytoskeleton formation (actin). These proteins appear to be involved in the resistance response to trichlorphon.

The COGITAT holeboard system as a valuable tool to assess learning, memory and activity in mice.

The comprehensive and stress-free assessment of various aspects of learning and memory is a prerequisite to evaluate mouse models for neuropsychiatric disorders such as Alzheimer's disease or attention deficit/hyperactivity disorder (ADHD). COGITAT is an automated holeboard system allowing simultaneous assessment of spatial working and reference-memory performance which we have adapted in this study to enable its usage with mice. The holeboard apparatus consists of an open-field chamber with a 25-hole floor insert, each hole being monitored by infrared light beams, located on three different levels, allowing the distinction between visits of holes, i.e. the animal reaches the bottom of the hole, or inspections, which means only superficial exploration of the hole. Across trials, animals learn a pattern of five baited holes. Here, we show that C57BL/6 mice readily acquire this task within 5 days when submitted to six trials per day. A number of individual parameters - overall exploratory activity, number of visits into or inspections of holes, number of baited, unbaited, or previously baited holes visited or inspected, reinspections of or revisits into any holes, number of pellets eaten, time to find pellets, and reference and working memory errors-are obtained simultaneously and results are immediately available after the end of each experiment. The muscarinic antagonist scopolamine impaired task performance, while the cognitive enhancer metrifonate (an acetylcholinesterase inhibitor) reduced error rates. Overall, our data indicate that this spatial learning task will be useful to characterize spatial memory in various genetic or pharmacological mouse models.

Insecticide resistance of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in mainland China.

BACKGROUND: In order to investigate the extent of resistance of oriental fruit fly, Bactrocera dorsalis (Hendel), which is a widespread pest throughout tropical, subtropical and temperate fruit crops, 25 populations of this insect were collected from 13 sites in mainland China in 2007 and 2008. In addition, resistant strains were established that showed increasing development of resistance. RESULTS: Compared with the susceptible strain, one population of B. dorsalis expressed high resistance (RR = 70.4-fold), 16 populations expressed medium resistance (11.5-fold < RR < 25.8-fold) and eight populations had low resistance or remained susceptible (1.4-fold < RR < 8.9-fold) to trichlorphon. As regards ß-cypermethrin, one population showed high resistance (RR = 44.0-fold), nine populations expressed medium resistance (12.2-fold < RR < 28.4-fold), 14 populations expressed low or minor resistance (3.0-fold < RR < 9.7-fold) and one population remained susceptible (RR = 1.1-fold). As regards avermectin, five populations had developed high resistance (44.3-fold < RR < 104-fold), seven populations expressed medium resistance (11.4-fold < RR < 38.6-fold) and three populations expressed low or minor resistance (3.5-fold < RR < 5.6-fold). A continuous resistance selection in the laboratory strain showed that the resistance ratios to trichlorphon, ß-cypermethrin and avermectin were 71.6-fold, 333-fold and 70.4-fold respectively. CONCLUSION: The data provided a comprehensive survey of insecticide resistance in Bactrocera dorsalis in mainland China. All results suggested that early resistance management programmes should be established for restoring the efficacy of pesticide-based control measures.

Selectivity of pesticides used in integrated apple production to the lacewing, Chrysoperla externa.

This research aimed to assess the toxicity of the pesticides abamectin 18 CE (0.02 g a.i. L-1), carbaryl 480 SC (1.73 g a.i. L-1), sulfur 800 GrDA (4.8 g a.i. L-1), fenitrothion 500 CE (0.75 g a.i. L-1), methidathion 400 CE (0.4 g a.i. L-1), and trichlorfon 500 SC (1.5 g a.i. L-1) as applied in integrated apple production in Brazil on the survival, oviposition capacity, and egg viability of the lacewing, Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) from Bento Gonçalves and Vacaria, Rio Grande do Sul State, Brazil. An attempt was made to study morphological changes caused by some of these chemicals, by means of ultrastructural analysis, using a scanning electronic microscope. Carbaryl, fenitrothion, and methidathion caused 100% adult mortality for both populations, avoiding evaluation of pesticides' effects on predator reproductive parameters. Abamectin and sulfur also affected the survival of these individuals with mortality rates of 10% and 6.7%, respectively, for adults from Bento Gonçalves, and were harmless to those from Vacaria at the end of evaluation. Trichlorfon was also harmless to adults from both populations. No compound reduced oviposition capacity. C. externa from Vacaria presented higher reproductive potential than those from Bento Gonçalves. In relation to egg viability, sulfur was the most damaging compound to both populations of C. externa. Ultrastructural analyses showed morphological changes in the micropyle and the chorion of eggs laid by C. externa treated with either abamectin or sulfur. The treatment may have influenced the fertilization of C. externa eggs and embryonic development. Sulfur was responsible for malformations in the end region of the abdomen and genitals of treated females. When applied to adults, abamectin, sulfur, and trichlorfon were harmless, while carbaryl, fenitrothion, and methidathion were harmful, according to the IOBC classification.

Resurrecting clinical pharmacology as a context for Alzheimer disease drug development.

Commercial priorities have been identified as negative factors in drug development. We trace the problem to inattention to sound clinical pharmacology practices. When properly applied, clinical pharmacology and associated drug development sciences can, hand in hand, facilitate success in commercial drug development.

In vitro reactivation of trichlorfon-inhibited butyrylcholinesterase using HI-6, obidoxime, pralidoxime and K048.

Trichlorfon is a specific inhibitor of cholinesterases. It was typically used as an insecticide; however, trichlorfon was described as useful for symptomatic treatment of Alzheimer's disease some years ago. The presented study is aimed at reactivation of trichlorfon-inhibited butyrylcholinesterase since this enzyme play an important role in Alzheimer's disease as deputy for acetylcholinesterase and furthermore it could be applied as a scavenger in case of overdosing. We used in vitro reactivation test for considering only reactivation efficacy of butyrylcholinesterase that is inhibited by trichlorfon and not reactivation of butyrylcholinesterase inhibited by trichlorfon metabolic products. Four reactivators were used: HI-6, pralidoxime, obidoxime, and K048. Although all of the reactivators seem to be effective at 1 mM concentration, a lower concentration was not able ensure sufficient reactivation. There was also an observed lowering of reactivation efficacy when butyrylcholinesterase was exposed to trichlorfon for a longer time interval.

The effect of levamisole on sheep trachea alone and with trichlorfon combination.

The aim of this study is to determine the effect of levamisole and levamisole-trichlorfon combination on isolated sheep trachea. Contraction was achieved with levamisole concentrations (10(-8)-10(-3) M) on tracheal strips of adult sheep (> 1 year of age). Pretreatment with trichlorfon (10(-7) M)-levamisole (10(-7) M) decreased the pD2 and E(max) of Ach when compared to levamisole pretreatment (p < or = 0.01). Pretreatment with levamisole (10(-7) M), decreased the E(max) (p < or = 0.01) and pD2 (p < 0.05) of bethanechol concentrations (10(-8)-10(-3) M) significantly. Pretreatment with atropine (10(-6) M) decreased the E(max) of levamisole (10(-4) M) significantly (p < 0.05). To conclude, levamisole acted mainly on the muscarinic receptors of the sheep trachea and this effect was partly inhibited by atropine. Adverse interaction was present between levamisole and trichlorfon.