Inhibitor Report for: Fenobucarb

Mixtures of organophosphate (OP) and carbamate (CB) insecticides are commonly detected in freshwater habitats. These insecticides inhibit the activity of acetylcholinesterase (AChE) and have potential to interfere with behaviors that may be essential for survival of species. Although the effects of individual anticholinesterase insecticides on aquatic species have been studied for decades, the combined toxicity of mixtures is still poorly understood. In the present study, we assessed whether pesticides in a mixture act in isolation (resulting in additive AChE inhibition) or whether components interact to produce either antagonistic or synergistic toxicity. Brain AChE inhibition in carp (Cyprinus carpio L.) exposed to a series of concentrations of the OP (chlorpyrifos, malathion and triazophos) as well as the CB (fenobucarb and carbosulfan) were measured. The concentration addition (CA) model and the isobole method were used to determine whether toxicological responses to binary mixtures of pesticides. In 50:50 % effect mixtures, the observed combined toxicity of chlorpyrifos and malathion was significantly higher than observed and was considered as synergistic. For equivalent dose mixtures, when chlorpyrifos mixed with fenobucarb or malathion, the observed toxicities were significantly higher than predicted, suggesting synergistic joint actions. The rest five binary combinations exhibited concentration additive or slight antagonistic joint actions. The CA model and the isobole method provided estimates of mixture toxicity that did not markedly underestimate the measured toxicity, therefore these methods are suitable to use in ecological risk assessments of pesticide mixtures.

Pardosa pseudoannulata is an important predatory enemy against insect pests, such as rice planthoppers and leafhoppers. In order to understand the insecticide selectivity between P. pseudoannulata and insect pests, two acetylcholinesterase genes, Pp-ace1 and Pp-ace2, were cloned from this natural enemy. The putative proteins encoded by Pp-ace1 and Pp-ace2 showed high similarities to insect AChE1 (63% to Liposcelis entomophila AChE1) and AChE2 (36% to Culex quinquefasciatus AChE2) with specific functional motifs, which indicated that two genes might encode AChE1 and AChE2 proteins respectively. The recombinant proteins by expressing Pp-ace1 and Pp-ace2 genes in insect sf9 cells showed high AChE activities. The kinetic parameters, Vmax and Km, of two recombinant AChE proteins were significantly different. The sensitivities to six insecticides were determined in two recombinant AChEs. Pp-AChE1 was more sensitive to all tested insecticides than Pp-AChE2, such as fenobucarb (54 times in Ki ratios), isoprocarb (31 times), carbaryl (13 times) and omethoate (6 times). These results indicated that Pp-AChE1 might be the major synaptic enzyme in the spider. By sequence comparison of P. pseudoannulata and insect AChEs, the key amino acid differences at or close to the functional sites were found. The locations of some key amino acid differences were consistent with the point mutation sites in insect AChEs that were associated with insecticide resistance, such as Phe331 in Pp-AChE2 corresponding to Ser331Phe mutation in Myzus persicae and Aphis gossypii AChE2, which might play important roles in insecticide selectivity between P. pseudoannulata and insect pests. Of course, the direct evidences are needed through further studies.

Seven field populations of mosquito Culex pipiens complex (Diptera: Culicidae) were collected from four provinces of China. The resistance status of larvae to dichlorvos, parathion, chlorpyrifos, fenobucarb (BPMC) and propoxur were determined by bioassays, disclosing that they were more resistant to organophosphate (moderate or low resistance) than to carbamate (low or no significant resistance) insecticides. Starch gel electrophoresis confirmed the presence and distribution of overproduced esterases B1, A2-B2, A8-B8 and A9-B9, the frequencies of which varied according to their regional origins. Electrophoretic polymorphism at four putatively neutral loci (got-1, got-2, pgi and pgm) showed that the overall genetic differentiation found across all populations was significantly large (Fst = 0.28, P < 10(-4)), and genetic exchange was slightly restricted by distance isolation (P = 0.018).

Mixtures of organophosphate (OP) and carbamate (CB) pesticides are commonly detected in freshwater ecosystems. These pesticides inhibit the activity of acetylcholinesterase (AChE) and have potential to interfere with behaviors that may be essential for the survival of species. Although the effects of individual anticholinesterase insecticides on aquatic species have been studied for decades, the neurotoxicity of mixtures is still poorly understood. In the present study, brain AChE inhibition in carp (Cyprinus carpio) exposed to a series of concentrations of the organophosphates (malathion and triazophos) as well as the carbamates (fenobucarb and carbosulfan) was measured. In equitoxic mixtures, the observed AChE activity inhibition of the malathion plus triazophos, and triazophos plus carbosulfan mixtures, was synergism. In equivalent concentration mixtures, the combination of malathion plus fenobucarb mixture conformed to synergism, while the observed AChE activity inhibition of the remaining pairings was less than additive. Single pesticide risk assessments are likely to underestimate the impacts of these insecticides on carps in aquatic environment where mixtures occur. Moreover, mixtures of pesticides that have been commonly reported in aquatic ecosystems may pose a more important challenge than previously anticipated.

Mixtures of organophosphate (OP) and carbamate (CB) insecticides are commonly detected in freshwater habitats. These insecticides inhibit the activity of acetylcholinesterase (AChE) and have potential to interfere with behaviors that may be essential for survival of species. Although the effects of individual anticholinesterase insecticides on aquatic species have been studied for decades, the combined toxicity of mixtures is still poorly understood. In the present study, we assessed whether pesticides in a mixture act in isolation (resulting in additive AChE inhibition) or whether components interact to produce either antagonistic or synergistic toxicity. Brain AChE inhibition in carp (Cyprinus carpio L.) exposed to a series of concentrations of the OP (chlorpyrifos, malathion and triazophos) as well as the CB (fenobucarb and carbosulfan) were measured. The concentration addition (CA) model and the isobole method were used to determine whether toxicological responses to binary mixtures of pesticides. In 50:50 % effect mixtures, the observed combined toxicity of chlorpyrifos and malathion was significantly higher than observed and was considered as synergistic. For equivalent dose mixtures, when chlorpyrifos mixed with fenobucarb or malathion, the observed toxicities were significantly higher than predicted, suggesting synergistic joint actions. The rest five binary combinations exhibited concentration additive or slight antagonistic joint actions. The CA model and the isobole method provided estimates of mixture toxicity that did not markedly underestimate the measured toxicity, therefore these methods are suitable to use in ecological risk assessments of pesticide mixtures.

Effects of dissolved pesticides on fish are widely described, but little is known about effects of pesticide-contaminated feeds taken up orally by fish. In integrated farms, pesticides used on crops may affect grass carp that feed on plants from these fields. In northern Vietnam, grass carp suffer seasonal mass mortalities which may be caused by pesticide-contaminated plants. To test effects of pesticide-contaminated feeds on health and bioaccumulation in grass carp, a net-cage trial was conducted with 5 differently contaminated grasses. Grass was spiked with 2 levels of trichlorfon/fenitrothion and fenobucarb. Unspiked grass was used as a control. Fish were fed at a daily rate of 20% of body mass for 10 d. The concentrations of fenitrothion and fenobucarb in pond water increased over time. Effects on fish mortality were not found. Fenobucarb in feed showed the strongest effects on fish by lowering feed uptake, deforming the liver, increasing blood glucose and reducing cholinesterase activity in blood serum, depending on feed uptake. Fenobucarb showed increased levels in flesh in all treatments, suggesting bio-concentration. Trichlorfon and fenitrothion did not significantly affect feed uptake but showed concentration-dependent reduction of cholinesterase activity and liver changes. Fenitrothion showed bioaccumulation in flesh which was dependant on feed uptake, whereas trichlorfon was only detected in very low concentrations in all treatments. Pesticide levels were all detected below the maximum residue levels in food. The pesticide-contaminated feeds tested did not cause mortality in grass carp but were associated with negative physiological responses and may increase susceptibility to diseases.

Pardosa pseudoannulata is an important predatory enemy against insect pests, such as rice planthoppers and leafhoppers. In order to understand the insecticide selectivity between P. pseudoannulata and insect pests, two acetylcholinesterase genes, Pp-ace1 and Pp-ace2, were cloned from this natural enemy. The putative proteins encoded by Pp-ace1 and Pp-ace2 showed high similarities to insect AChE1 (63% to Liposcelis entomophila AChE1) and AChE2 (36% to Culex quinquefasciatus AChE2) with specific functional motifs, which indicated that two genes might encode AChE1 and AChE2 proteins respectively. The recombinant proteins by expressing Pp-ace1 and Pp-ace2 genes in insect sf9 cells showed high AChE activities. The kinetic parameters, Vmax and Km, of two recombinant AChE proteins were significantly different. The sensitivities to six insecticides were determined in two recombinant AChEs. Pp-AChE1 was more sensitive to all tested insecticides than Pp-AChE2, such as fenobucarb (54 times in Ki ratios), isoprocarb (31 times), carbaryl (13 times) and omethoate (6 times). These results indicated that Pp-AChE1 might be the major synaptic enzyme in the spider. By sequence comparison of P. pseudoannulata and insect AChEs, the key amino acid differences at or close to the functional sites were found. The locations of some key amino acid differences were consistent with the point mutation sites in insect AChEs that were associated with insecticide resistance, such as Phe331 in Pp-AChE2 corresponding to Ser331Phe mutation in Myzus persicae and Aphis gossypii AChE2, which might play important roles in insecticide selectivity between P. pseudoannulata and insect pests. Of course, the direct evidences are needed through further studies.

To determine sensitivity to fenitrothion (MEP), fenobucarb (BPMC) and imidacloprid (IMI) in each larval instar of Cheumatopsyche brevilineata, we carried out 48-hr acute toxicity tests using strains M and K originating from an urban river and an irrigation canal, respectively. First instar was the most sensitive stage to all three insecticides whereas fifth instar was the most insensitive in both strains. Larvae of strain K were significantly more insensitive to MEP through larval growth and to BPMC in second or later instars than larvae of strain M; however, sensitivity to IMI did not differ significantly between strains.

To investigate insecticide resistance levels and dynamic changes of carboxylesterase polymorphism with time in resistant populations of mosquitoes in the Culex pipiens complex, four field populations were collected in September 2006. The resistance levels of fourth-instar larvae to organophosphate (dichlorvos, parathion, and chlorpyrifos), carbamate (fenobucarb and propoxur), and pyrethroid (permethrin and tetramethrin) insecticides were determined by bioassay. Larvae had a low but significant resistance to organophosphate and carbamate insecticides but no significant resistance to pyrethroid insecticides. Starch gel electrophoresis revealed the presence of the overproduced esterases B1, A2-B2, A8-B8, A9-B9, and All-B11. The frequency of each overproduced esterases varied depending on its regional localities. Compared with previous surveys, the polymorphism of amplified esterase alleles in Beijing populations increased, although the resistance level to organophosphate insecticides declined.

Seven field populations of mosquito Culex pipiens complex (Diptera: Culicidae) were collected from four provinces of China. The resistance status of larvae to dichlorvos, parathion, chlorpyrifos, fenobucarb (BPMC) and propoxur were determined by bioassays, disclosing that they were more resistant to organophosphate (moderate or low resistance) than to carbamate (low or no significant resistance) insecticides. Starch gel electrophoresis confirmed the presence and distribution of overproduced esterases B1, A2-B2, A8-B8 and A9-B9, the frequencies of which varied according to their regional origins. Electrophoretic polymorphism at four putatively neutral loci (got-1, got-2, pgi and pgm) showed that the overall genetic differentiation found across all populations was significantly large (Fst = 0.28, P < 10(-4)), and genetic exchange was slightly restricted by distance isolation (P = 0.018).

Dual counter-current chromatography (dual CCC) has been successfully applied to rapid sample preparation for the simultaneous determination of residual carbaryl, fenobucarb and methomyl in vegetable oil and citrus fruit. The citrus fruit samples were extracted with n-hexane solution containing stable isotopically labeled internal standards (methomyl-d3, fenobucarb-d3 and carbaryl-d9), and applied to dual CCC using a two-phase solvent system of n-hexane-acetonitrile to purify the carbamate pesticides from aliphatic sample matrix. The coiled column was rotated at 420 rpm, the lower mobile phase was introduced through the head toward the tail, and the upper mobile phase in the opposite direction. Due to the high partition efficiency of dual CCC, the lower phase fraction collected from 2 to 5 min after injection could be subjected to flow-injection tandem mass spectrometry directly after concentration. Repetitive sample injection can be performed at high reproducibility without a risk of contamination from the compounds retained in the column.

With the expansion of agricultural areas within the Mekong River Delta in Vietnam, a concurrent, dramatic increase has occurred in agrochemical usage. To date, little consideration has been given to the negative impacts of this agricultural activity on the aquatic resources of the region. Both acute toxicity and subacute effects on brain cholinesterase (ChE) of two of the most commonly used insecticides, diazinon and fenobucarb, on adult native snakehead (Channa striata) were evaluated in a static, nonrenewable system, the environmental parameters of which, such as dissolved oxygen, water temperature, and pH, fluctuated similarly to field conditions. Four levels of insecticides, from 0.008 to 0.52 mg/L (for diazinon) and from 0.11 to 9.35 mg/L (for fenobucarb), were tested to assess the effects on the brain ChE activity of the snakehead up to 30 and 10 d for diazinon and fenobucarb, respectively. Diazinon was highly toxic to this fish species, with a 96-h median lethal concentration (LC50) of only 0.79 mg/L, and it also caused long-term ChE inhibition, with activity still significantly inhibited by 30% after 30 d for the three highest concentrations. Fenobucarb was less toxic to this species, with a 96-h LC50 of 11.4 mg/L. Fenobucarb caused more rapid ChE inhibition but also rapid recovery. The results of the present study indicate an urgent need to regulate the usage of these pesticides in the Mekong River Delta.

The dissipation rates of dimethoate, fenobucarb, flutolanil, simazine, prometryn and alachlor were compared among an andosol and a gray lowland soil subjected to different fertilizing practices over a 20 year period. The rate constants for dissipation of most pesticides per biomass carbon, biomass nitrogen and esterase activity among the plots in each soil were less variable than the corresponding rate constants, indicating that the dissipation depended on microbial amount and activity. The rate constants in the gray lowland soil were similar to or greater than those in the andosol, despite the smaller values of microbial amount and activity in the former. This is due to the larger water soluble fraction/acetone soluble fraction ratios in the former. The long-term successive applications of organic fertilizers were less effective in the dissipation for the gray lowland soil than the andosol. This is likely to result from a less effective accumulation of microbial biomass in the former.