Variation in the sensitivity of Callosobruchus (Coleoptera: Bruchidae) acetylcholinesterase to the organophosphate insecticide malaoxon: effect of species, geographical strain and food type

BACKGROUND: Bruchid beetles, Callosobruchus species, are serious pests of economically important grain legumes; their activity in stores is often controlled by the use of synthetic insecticides. Esterases are known to be involved in insecticide resistance in insects. However, there is a dearth of information on esterase activity in the genus Callosobruchus. In this study, the effect of species, geographical strain and food type on the variation in acetylcholinesterase (AChE) activity and its inhibition by malaoxon (malathion metabolite) was investigated using an in vitro spectrophotometric method. RESULT: AChE activity varied significantly among species and strains and also among legume type used for rearing them. Generally, irrespective of species, strain or food type, the higher the AChE activity of a population, the higher is its inhibition by malaoxon. C. chinensis had the highest AChE activity of the species studied, and in the presence of malaoxon it had the lowest remaining AChE activity, while C. rhodesianus retained the highest activity. CONCLUSION: A first‐hand knowledge of AChE activity in regional Callosobruchus in line with the prevailing food types should be of utmost importance to grain legume breeders, researchers on plant materials for bruchid control and pesticide manufacturer/applicators for a robust integrated management of these bruchids. Copyright © 2012 Society of Chemical Industry     

生化法快速测定蔬菜中农药残毒

用自行研制的便携式农药残毒速测仪 ,对蔬菜中残留农药进行生化检测。依据有机磷和氨基甲酸酯类农药对乙酰胆碱酯酶的活性有抑制的原理 ,绘制农药残毒———吸光度标准曲线 ,读出吸光度即得到相应的农药浓度。     

感染Bt的3龄思茅松毛虫中肠组织病理及血淋巴酶活性分析

从昆明安宁采集思茅松毛虫茧至室内羽化、孵化,将发育至3龄幼虫作为试虫,基于Probit回归法对其进行Bt制剂毒力测定分析,利用最佳检测时间点的致死中浓度（LC₅₀）对其进行感染,在感染后不同时间显微镜检中肠组织病变,同时测定血淋巴的酚氧化酶（PO）、羧酸酯酶（CarE）和乙酰胆碱酯酶（AChE）等活性,探究苏云金杆菌（Bt）对思茅松毛虫幼虫中肠组织和血淋巴酶活性的影响,分析其作用机制。结果表明：48 h是Bt制剂毒力测定的最佳检测时间,其LC₅₀为13.024 IU/μL;中肠典型病变包括：柱状细胞伸长变形、杯状细胞数量明显增多及其胞腔显著扩大、围食膜逐渐消失等;Bt感染后幼虫血淋巴CarE、AChE和PO的酶活性显著升高,其中AChE最为明显。     

The impact of carbofuran on acetylcholinesterase activity in Anisakis simplex larvae from Baltic herring

The aim of this study was to investigate the impact of carbofuran, a carbamate pesticide on the enzymatic activity of acetylcholinesterase (AChE) in the larvae of the herring nematode parasite Anisakis simplex. A. simplex larvae collected from herring were exposed to carbofuran in vivo at concentrations of 50, 100, 500 and 1000 μg/l, for 24, 48 and 72 h, at a temperature of 4 °C. Generalized Linear Models (GLM) were applied to analyze the relationship between AChE activity and carbofuran concentration, the time of exposure and the biological parameters of the host. The results indicate that A. simplex larvae have a high threshold of sensitivity to carbofuran. The average enzymatic activity was higher in parasites obtained from male hosts, when compared with female hosts. These data suggest that host sex-dependent biological processes may also influence AChE enzymatic activity in parasites.     

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).     

New cholinesterase inhibiting bisbenzylisoquinoline alkaloids from Abuta grandifolia

The phytochemical study of the stem bark and wood of Abuta grandifolia (Mart.) Sandwith led to the identification of four bisbenzylisoquinoline alkaloids (BBIQs), namely (R,S)-2 N-norberbamunine (1), (R,R)-isochondodendrine (2), (S-S)-O4″-methyl, Nb-nor-O6'-demethyl-(+)-curine (3), and (S-S)-O4″-methyl, O6'-demethyl-(+)-curine (4), together with the aporphine alkaloid R-nornuciferine (5), all obtained by countercurrent distribution separation (CCD) and identified on the basis of their spectroscopic data. Alkaloids 3 and 4 were new. All the isolated compounds were tested for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities. 1 was the most active against AChE, whereas 3 and 4 were the most potent against BChE. Interestingly, all tested alkaloids are more potent against BChE than against AChE. This selectivity of cholinesterase (ChE) inhibition could be important in order to speculate on their potential therapeutic relevance.     

杀虫药剂混用对棉铃虫乙酰胆碱酯酶和羧酸酯酶的联合抑制作用研究

选用辛硫磷、对硫磷、敌敌畏和氧乐果4种有机磷类杀虫剂以及灭多威和甲萘威两种氨基甲酸酯类杀虫剂分别以1∶1、1∶3和3∶1的比例混用,以棉铃虫Helicoverpa armigera为试虫,分别测得单剂和混剂对其乙酰胆碱酯酶(AChE)和羧酸酯酶(CarE)的抑制中浓度(I50),以联合抑制系数作为增效作用的参考指标进行了比较。研究发现,两种药剂不同配比的混剂对同种酯酶的联合抑制作用也往往不同。对AChE和CarE同时具有抑制作用的只有其中的对硫磷＋甲萘威(1∶3)、辛硫磷+对硫磷(1∶3)和灭多威+氧乐果(1∶3),而甲萘威+敌敌畏(1∶3)、甲萘威＋氧乐果(1∶1)、辛硫磷+甲萘威(1∶1)、对硫磷+氧乐果(1∶1)、对硫磷+氧乐果(1∶3)、敌敌畏+氧乐果(1∶3)、灭多威+对硫磷(1∶1) 7种混剂对AChE和CarE都具有拮抗作用。     

Toxicological and biochemical characterization of coumaphos resistance in the San Roman strain of Boophilus microplus (Acari: Ixodidae)

The San Roman strain of the southern cattle tick, Boophilus microplus, collected from Mexico was previously reported to have a high level of resistance to the organophosphate acaricide coumaphos. An oxidative detoxification mechanism was suspected to contribute to coumaphos resistance in this tick strain, as coumaphos bioassay with piperonyl butoxide (PBO) on larvae of this resistant strain resulted in enhanced coumaphos toxicity, while coumaphos assays with PBO resulted in reduced toxicity of coumaphos in a susceptible reference strain. In this study, we further analyzed the mechanism of oxidative metabolic detoxification with synergist bioassays of coroxon, the toxic metabolite of coumaphos, and the mechanism of target-site insensitivity with acetylcholinesterase (AChE) inhibition kinetics assays. Bioassays of coroxon with PBO resulted in synergism of coroxon toxicity in both the San Roman and the susceptible reference strains. The synergism ratio of PBO on coroxon in the resistant strain was 4.5 times that of the susceptible strain. The results suggested that the cytP450-based metabolic detoxification existed in both resistant and susceptible strains, but its activity was significantly enhanced in the resistant strain. Comparisons of AChE activity and inhibition kinetics by coroxon in both susceptible and resistant strains revealed that the resistant San Roman strain had an insensitive AChE, with a reduced phosphorylation rate, resulting in a reduced bimolecular reaction constant. These data indicate a mechanism of coumaphos resistance in the San Roman strain that involves both insensitive AChE and enhanced cytP450-based metabolic detoxification.     

松油烯-4-醇对家蝇体内几种酶系的影响

测定了松油烯-4-醇对家蝇成虫的毒杀活性及其体内几种酶系活性的影响.结果表明:松油烯-4-醇对家蝇具有较强的触杀作用,24h的LD50为23.91μg/虫;对家蝇的致毒症状可明显地分为兴奋、痉挛、麻痹、死亡或复苏等几个阶段;对Na ,K -ATPase具有较强的抑制作用,在兴奋期、痉挛期、麻痹期和复苏期,酶抑制率分别为23.2l%、20.27%、61.84%和58.35%;离体条件下,对Na ,K -ATPase的I50为0.8379×103μg·mL-1;对AChE有一定的抑制作用;可激活酯酶,在复苏期,酶比活力为对照的1.42倍.     

Target site insensitivity and mutational analysis of acetylcholinesterase from a carbofuran-resistant population of Colorado potato beetle, Leptinotarsa decemlineata (Say)

The BERTS strain of Colorado potato beetle (CPB) was found to be highly resistant to N-methyl carbofuran but relatively susceptible to azinphosmethyl. N-Methyl carbofuran resistance was found to correlate well with acetylcholinesterase (AChE) insensitivity. In becoming resistant to N-methyl carbofuran, the AChE of the BERTS strain became more sensitive to N-propyl carbofuran inhibition. This negative cross-insensitivity correlated well to the increased relative toxicity of the BERTS strain to N-propyl carbofuran compared to the susceptible SS strain. BERTS beetles were sorted into BERTS-R and BERTS-S substrains using their AChE activity profiles. Sequence comparisons of AChE cDNAs from the two substrains revealed the presence of the point mutation that results in the S291G substitution previously found in the AChE of the azinphosmethyl-resistant AZ-R strain of CPB. A novel mutation present only in BERTS-S CPB, however, resulted in an additional I392T substitution in the AChE and apparently reverses the resistance conferring properties of the S291G substitution.