Comparative studies of acetylcholinesterases purified from two field populations of the oriental migratory locust (Locusta migratoria manilensis): implications of insecticide resistance

Acetylcholinesterase (AChE) was purified by affinity chromatography from two populations of the oriental migratory locust, Locusta migratoria manilensis (Meyen), collected from Huanghua and Pingshan Counties, Hebei Province of China. The purification factors and yields were 1661-fold and 19.3%, respectively, for the Huanghua population, and 3897-fold and 39.6% for the Pingshan population. Both the purification factor and yield were significantly lower in the Huanghua population than in the Pingshan population. AChE activity was almost completely inhibited by 10⁻⁶ M eserine and BW284C51, but ?5.8% of AChE activity was inhibited by ethopropazine at the same concentration, suggesting that purified AChE from either population was a typical insect AChE. However, AChE purified from the Huanghua population was 62-, 2.0-, and 1.6-fold less sensitive to inhibition by the three organophosphate compounds, chlorpyrifos oxon, demeton-S-methyl, and paraoxon, respectively, than that from the Pingshan population. Significantly lower purification factor and low yield associated with reduced sensitivity of AChE to inhibition by the organophosphates indicated that AChE purified from the Huanghua population was biochemically and pharmacologically different from that of the Pingshan population. Reduced sensitivity of AChE appeared to contribute to organophosphate resistance in the locust from Huanghua County, where insecticides have commonly been used to manage outbreaks of the locust.     

Tissue-specific antioxidative and neurotoxic responses to diazinon in Oreochromis niloticus

The purpose of this study was to evaluate oxidative stress and neurotoxic potential of organophosphorus (OP) insecticide diazinon in the sentinel freshwater fish, Oreochromis niloticus. Antioxidant and acetylcholinesterase (AChE) enzyme activities and malondialdehyde (MDA) and protein levels were measured spectrophotometrically in gill, kidney, alimentary tract, and muscle tissues of fish treated with sub-lethal diazinon concentrations for 1, 7, 15, and 30 days. Dose-dependent inhibitions of AChE were observed in all the experimental fish. On the contrary of alimentary tract, MDA levels were elevated in kidney and muscle and gill was not affected. AChE and MDA levels intercorrelated in kidney and muscle tissues. Diazinon had increased superoxide dismutase (SOD) activities in all the tissues, while kidney was the most affected tissue. Tissue-specific alterations were observed on catalase (CAT) and glutathione peroxidase (GPx) activities; however, the activities were not changed in gill and muscle tissues for GPx and in gill, muscle, and kidney tissues for CAT. Protein levels decreased in kidney, muscle, and alimentary tract, while increased in gill and alimentary tract in 15 days. With respect to these results, diazinon has oxidative and neurotoxic potentials in O. niloticus. Observed changes with diazinon treatment were generally tissue-specific and dose-dependent.     

Concentration related responses of chlorpyriphos in antioxidant, anaerobic and protein synthesizing machinery of the freshwater fish, Heteropneustes fossilis

Impacts of chlorpyriphos (2, 4, 6 ppm) on the profiles of antioxidant (catalase) and anaerobic (lactate dehydrogenase) enzymes and other macromolecular contents (DNA, RNA, protein) of various tissues of the freshwater fish, Heteropneustes fossilis were studied. Chlorpyriphos significantly decreased the specific activity of catalase (CAT) and lactate dehydrogenase (LDH) in the brain, liver, gill and skeletal muscle of the fish. The reduction in specific activity might be due to the binding of chlorpyriphos or its metabolites with the enzyme molecules or affecting the synthesis and/or degradation of the enzymes. Like enzymes, the DNA, RNA and protein contents decreased in the brain, liver, gill and skeletal muscle of the fish as a function of increase in chlorpyriphos concentrations (2-6 ppm). The chlorpyriphos-induced reduction in these biochemical constituents might be because of alteration in their turnover (synthesis/degradation) in different tissues. The maximum effects on CAT, LDH, DNA, RNA and protein were obtained in response to 6 ppm chlorpyriphos. The present findings suggest chlorpyriphos concentration related impairment in antioxidative, anaerobic and protein synthesizing capacity of the fish. Therefore, the use of higher concentrations of chlorpyriphos should be avoided to protect the health of economically important freshwater food fish.     

Potential environmental consequences of administration of ectoparasiticides to sheep

Sheep ectoparasiticides, which include the synthetic pyrethroids, the organophosphates, the 'insect'-growth regulators, the formamidines and the spinocyns, enter into the environment primarily through disposal of dip or fleece scours, as well as with contaminated faeces and urine. Due to the large quantities of spent dip, risks associated with environmental contamination are high. Synthetic pyrethroids and organophosphates pose risks to dung, soil and aquatic fauna; concerns over potential ecotoxicity to vertebrates and invertebrates have resulted in the cessation of their use in many countries. There is very little information regarding the ecotoxicity of 'insect'-growth regulators, formamidines or spinocyns, with no studies focussing on sheep. Here, the impact of sheep ectoparasiticides is discussed in terms of their potential to enter into the environment, their toxicity and their impact on ecosystem functioning. Where there are no data for excretion or toxicity of the ectoparasiticides used in sheep production, examples to demonstrate potential impacts are taken from laboratory ecotoxicity tests and the cattle literature, as well on work with foliar insecticides. Future research priorities are suggested to allow assessment of the environmental consequences of sheep ectoparasiticide treatments, which are essential for future sustainable sheep production.     

Evaluation of thiacloprid and clothianidin (neonicotinoids) as alternative to fenthion (organophosphate) for control of Mediterranean fruit fly (Diptera: Tephritidae) in deciduous fruit orchards

Fenthion (organophosphate) was used in Australia to target adults, eggs and larvae of Mediterranean fruit fly (Medfly), Ceratitis capitata Wiedmann (Diptera: Tephritidae) in deciduous fruits. With its removal from use in 2015, clothianidin and thiacloprid (neonicotinoids) were evaluated for any adulticide, ovicide or larvicide (eggs, larvae) activity against Medfly in a series of laboratory and field-cage experiments in peach and nectarine. We included fenthion in all experiments as a positive control. As an adulticide, fenthion had the greatest residual toxicity on fruit, with 7 d old residues killing 44% of adults within 48 h compared to thiacloprid (17%) and clothianidin (30%). When applied as a larvicide to naturally infested peaches, none of the insecticides was 100% effective, though fewest pupae (1.18 ± 0.19 pupae/fruit) (indirect measurement of larval survival) were produced when treated with fenthion, followed by clothianidin (3.40 ± 0.33 pupae/fruit) and thiacloprid (6.88 ± 0.34 pupae/fruit); with 11.35 ± 0.30 pupae/fruit in control (water). In peaches treated 48 h post-infestation, fenthion was most effective in reducing numbers of pupae and adult emergence and with higher percentage of deformed adults, with thiacloprid=clothianidin. In nectarines treated 48 h post-infestation, fenthion was most effective in reducing numbers of pupae and adult emergence as well as with higher percentage of deformed adults, followed by clothianidin. Insecticides were least effective when applied as a preventative 24 h or 7 d prior to infestation, with fenthion more effective than clothianidin. Whilst our study shows that neither clothianidin nor thiacloprid are equivalent to fenthion, they are acceptable substitutes to growers for use for Medfly in deciduous fruits given that fenthion is no longer available to growers in Australia. We recommend that growers incorporate other controls to target the adult stage rather than rely only on new insecticides.     

The effects of organophosphate insecticide diazinon on malondialdehyde levels and myocardial cells in rat heart tissue and protective role of vitamin E

Diazinon is an organophosphate insecticide has been used in agriculture and domestic for several years. Vitamin E (200 mg/kg, twice a week), diazinon (10 mg/kg, per day), and vitamin E (200 mg/kg, twice a week)+diazinon (10 mg/kg, per day) combination was given to rats orally via gavage for 7 weeks. Body and heart weights, malondialdehyde (MDA) level in heart tissue and ultrastructural changes in myocardial cells were investigated at the end of the 1st, 4th, and 7th weeks comparatively with control group. When diazinon-treated group was compared to control group body and heart weights were decreased significantly at the end of the 4th and 7th weeks. It was observed that, at the end of 1st, 4th, and 7th weeks there was a statistically significant increase in MDA levels when diazinon- and vitamin E +diazinon-treated groups were compared to control group. While at the end of the 1st week statistically significant changes were not being observed, at the end of 4th and 7th weeks statistically significant decrease was detected in MDA levels when vitamin E+diazinon-treated group was compared to diazinon-treated group. In our electron microscopic investigations, while vacuolization and swelling of mitochondria myocardial cells of diazinon-treated rats were being observed, swelling of several mitochondria were observed in vitamin E+diazinon-treated rats. We conclude that vitamin E reduces diazinon cardiotoxicity, but vitamin E does not protect completely.     

Detection of resistance-associated point mutations of organophosphate-insensitive acetylcholinesterase in the olive fruit fly, Bactrocera oleae (Gmelin)

We have recently identified two resistance-associated point mutations of organophosphate (OP)-insensitive acetylcholinesterase in the olive fruit fly Bactrocera oleae, the most important olive orchard pest world-wide. We have developed simple PCR-restriction fragment length polymorphism assays for each mutation, utilising an AccI restriction site created by Ile214Val, and a BssHII restriction site destroyed by a neutral change always accompanying the second mutation Gly488Ser. Samples from Greece homozygous for both mutations proved the most insensitive to dimethoate. The frequencies of these mutations in field-collected samples from several countries were investigated. Ninety-three percent of samples from Greece and Albania, where OPs have been extensively used in B. oleae control, were homozygous for both mutations. Resistance-associated alleles were detected at lower frequencies, but still with both mutations in conjunction in the majority of cases, in western Mediterranean countries with limited use of OPs. Samples from South Africa, however, did not have either of the resistance-associated mutations. The double mutation haplotype clearly confers a strong selective advantage in field populations of B. oleae exposed to OPs.     

液液萃取-固相萃取气相色谱法测定火腿中有机磷农药残留量

[目的]建立火腿中多种有机磷农药残留量同时测定的方法。[方法]火腿中有机磷农药经乙腈提取后,用乙腈-正己烷液液萃取除去脂类和色素,再经石墨碳黑固相萃取小柱净化,气相色谱-火焰光度检测器(GC-FPD)测定。[结果]16种有机磷农药在DB-17、DB-1色谱柱上均获得良好分离,在0.05~1.00μg/m L范围内,农药残留量色谱分析的线性关系良好,满足定量分析要求,加标水平为0.08和0.20 mg/kg时,农药的平均回收率在71.96%~117.21%,变异系数为1.73%~12.84%。方法的检出限为0.005 3~0.011 8 mg/kg。[结论]该方法具有前处理简单、净化效果好、方法准确、灵敏度高、杂质干扰少的特点,适合火腿中有机磷农药残留量的分析。