Oxidative stress and locomotor behaviour response as biomarkers for assessing recovery status of mosquito fish, Gambusia affinis after lethal effect of an organophosphate pesticide, monocrotophos

Recovery study was performed at regular intervals to establish the time course of 50% and 100% recovery in neurotransmitter enzyme (acetylcholinesterase, AChE, EC 3.1.1.7) and locomotor behaviour response of mosquito fish, Gambusia affinis exposed to lethal concentration (20.49 mg L⁻¹) of an organophosphorous pesticide, monocrotophos (MCP) for 96 h. In vitro AChE activity studies indicated that MCP could cause 50% inhibition (I₅₀) at 10.2 × 10⁻⁵ M. A positive correlation was observed between brain AChE activity and swimming speed during the recovery study. Also, the recovery response of the antioxidant enzymes superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) as well as lipid peroxidation (LPO) as biomarkers of oxidative stress were assessed in viscera of G. affinis. The results showed that the MCP besides its inhibitory effect on target enzyme AChE activity and induction in antioxidant enzyme activities as a characteristic of oxidative stress, which can be used as biomarkers in the pesticide contaminated aquatic streams.     

The relative toxicities of insecticides to earthworms of the Pheretima group (Oligochaeta)

An artificial soil test was used to determine the LC50 values of carbaryl, chlorpyrifos, imidacloprid, cyfluthrin and fipronil against earthworms of the Pheretima group. For a 24-h interval, carbaryl was the most toxic to earthworms (LC50 = 77 mg kg-1), followed by imidacloprid (155 mg kg-1), cyfluthrin (351 mg kg-1), chlorpyrifos (390 mg kg-1) and fipronil (> 8550 mg kg-1) as the least toxic. For the 48-h and 7-day intervals, imidacloprid was the most toxic to earthworms (LC50 = 5 mg kg-1 and 3 mg kg-1 respectively), followed by carbaryl (16 mg kg-1; 9 mg kg-1), cyfluthrin (128 mg kg-1; 110 mg kg-1), chlorpyrifos (330 mg kg-1; 180 mg kg-1) and the least toxic was fipronil (> 8550 mg kg-1 both intervals). The surface application rates required to achieve these values are compared with the rates recommended for the control of turfgrass pests.     

河南信阳、漯河黏虫种群对6种杀虫剂的敏感性

采用浸叶法测定了信阳和漯河两地黏虫种群对甲氨基阿维菌素苯甲酸盐(甲维盐)和氯虫苯甲酰胺等6种杀虫剂的敏感性。结果表明:信阳种群对6种药剂的敏感性由高到低依次为:甲维盐、氯虫苯甲酰胺、辛硫磷、高效氯氰菊酯、毒死蜱、高效氯氟氰菊酯,LC50分别为:0.035、0.230、3.566、3.716、5.025、13.315 mg/L;漯河种群的敏感性由高到低依次为:甲维盐、氯虫苯甲酰胺、毒死蜱、辛硫磷、高效氯氰菊酯、高效氯氟氰菊酯,LC50分别为:0.053、0.734、3.081、3.413、13.570、25.067 mg/L。以毒死蜱为标准药剂,甲维盐对信阳和漯河3龄黏虫幼虫都表现出极高的活性,相对毒力指数分别达到143.571和58.132。     

Effect of methyl parathion and chlorpyrifos on certain biomarkers in various tissues of guppy fish, Poecilia reticulata

Tests of acute toxicity were performed on the most common species of aquarium fish, Poecilia reticulata. Guppies (P. reticulata) were exposed to progressive concentrations of methyl parathion (MP) and chlorpyrifos (CPF); a semi-static method according to guidelines of OECD was used. Tests of acute toxicity were conducted using 10 fish for each separate concentration and for the control group. The results were subjected to probit analysis to determine the 96 h LC₅₀ values. The 96 h LC₅₀ values of MP and CPF to P. reticulata were 8.48 ppm/L (5.98–10.89) and 0.176 ppm/L (0.313–0.224) respectively. In addition, behavioral changes at each concentration were observed for the individual fish. Fish were exposed for 96 h to different sublethal concentrations of MP and CPF (¼ LC₅₀, 1/8 LC₅₀ and 1/10 LC₅₀) and their oxidative stress-induction potential was estimated in brain, liver and gills of fish. MDA content is induced in all tissues but maximum rise was observed in gills (161% and 153% for MP and CPF respectively). With regard to antioxidant defense system (ADS), GSH level decreased in the brain, liver and gills of tissues of MP treated fishes (22%, 6% and 13% respectively) and showed increase in brain and gills CPF treated (23% and 21% respectively). CAT, GST, GR and SOD levels fluctuated in all treatment groups relative to the control. Brain AChE showed dose-dependent inhibition in fish exposed to the higher concentrations reached 45% and 66% for MP and CPF respectively. Collective findings demonstrated that pesticide exposure of fish induced an increase in MDA and fluctuated ADS along with inhibited AChE. These findings may be used as valuable biomarkers for evaluation of water pollution.     

几种常用杀虫剂对瓜蚜的毒力和对乙酰胆碱酯酶的抑制作用

用叶片药膜法测定了4种常用杀虫剂氧乐果、辛硫磷、三唑磷、吡虫啉对呼和浩特市郊蔬菜基地温室黄瓜上发生的瓜蚜的毒力。结果表明,瓜蚜对吡虫啉最敏感,LC50 仅为4.203 mg/L,氧乐果、辛硫磷、三唑磷对瓜蚜的LC50分别为21.70、38.86、43.30 mg/L。研究明确了3种有机磷杀虫剂对瓜蚜体内乙酰胆碱酯酶AChE均有明显的抑制作用,并随着抑制时间的延长,抑制率增加。I50值分别为氧乐果9.6×10-6mol/L,辛硫磷11.4×10-6mol/L,三唑磷17.1×10-6mol/L。     

密褐褶孔菌Gloeophyllum trabeum对几种杀虫剂跟踪反应及对水坝白蚁诱杀效果的影响

测定了毒死蜱、氰戊菊酯、阿维菌素和氟虫腈等 4种杀虫剂与不同饵料混合后对家白蚁 Coptotermes formosanus和黄肢散白蚁 Reticulitermes flaviceps的口服毒性。在此基础上比较了各种饵料及由它们配成的毒饵对黄肢散白蚁的引诱力。结果表明 ,白蚁对甘蔗粉的喜好程度远超过对淀粉和松木粉。而受密褐褶孔菌 Gloeop hyllum trabeum侵染的甘蔗粉对白蚁的引诱力又远超过未受侵染的甘蔗粉。 4种杀虫剂中除氰戊菊酯外 ,另 3种与受侵染甘蔗粉制成的毒饵均能够在不损失杀白蚁活性的同时保留其对白蚁的引诱力。其中毒死蜱毒饵在为期30 d的水坝白蚁诱杀防治中取得最高 (6 6 .7% )的防治效果。     

Degradation of bifenthrin,chlorpyrifos and imidacloprid in soil and bedding materials at termiticidal application rates

Organophosphorus, pyrethroid and chloronicotinyl insecticides have been used to control termites in building structures in recent years. We investigated the degradation behaviour of three insecticides (bifenthrin, chlorpyrifos and imidacloprid) at termiticidal application rates under standard laboratory conditions (25 °C, 60% field moisture capacity and darkness) for 24 months. The study was carried out on one soil and two bedding materials (sand-dolomite and quarry sand), which are commonly used under housing in Australia. Experiments were also conducted to examine the effect of soil moisture on the degradation of these insecticides. Insecticide residues in the samples collected at different days after application were measured by high performance liquid chromatography (HPLC). The rate of degradation of bifenthrin and imidacloprid insecticides was adequately described by a first-order kinetic model (r² = 0.93–0.97). However, chlorpyrifos degradation was biphasic, showing an initial faster degradation followed by a slower rate. Therefore, the degradation data during the slower phase only (after a two-month period) followed the first-order law (r² = 0.95). Soil moisture had little effect on degradation of imidacloprid and bifenthrin. Among the three insecticides, bifenthrin and imidacloprid were most stable and chlorpyrifos the least. Chlorpyrifos showed a major loss (75–90%) of residue during the 24 months incubation period. In the bedding materials, simultaneous accumulation of the primary metabolite of chlorpyrifos, TCP (3,5,6-trichloro-2-pyridinol) was observed. Hydrolysis appeared to have caused the observed rapid loss of chlorpyrifos, especially in the highly alkaline bedding materials (sand-dolomite and quarry sand). © 1999 Society of Chemical Industry     

Resistance mechanisms and associated mutations in acetylcholinesterase genes in Sitobion avenae (Fabricius)

Wheat aphid, Sitobion avenae (fabricius), is one of the most important wheat pests and has been reported to be resistant to commonly used insecticides in China. To determine the resistance mechanism, the resistant and susceptible strains were developed in laboratory and comparably studied. A bioassay revealed that the resistant strain showed high resistance to pirimicarb (RR: 161.8), moderate reistance to omethoate (32.5) and monocrotophos (33.5), and low resistance to deltamethrin (6.3) and thiodicarb (5.5). A biochemistry analysis showed that both strains had similar glutathione-S-transferase (GST) activity, but the resistant strain had 3.8-fold higher esterase activity, and its AChE was insensitive to this treatment. The I₅₀ increased by 25.8-, 10.7-, and 10.4-folds for pirimicarb, omethoate, and monocrotophos, respectively, demonstrating that GST had not been involved in the resistance of S. avenae. The enhanced esterase contributed to low level resistance to all the insecticides tested, whereas higher resistance to pirimicarb, omethoate, and monocrotophos mainly depended on AChE insensitivity. However, the AChE of the resistant strain was still sensitive to thiodicarb (1.7-fold). Thus, thiodicarb could be used as substitute for control of the resistant S. avenae in this case. Furthermore, the two different AChE genes cloned from different resistant and susceptible individuals were also compared. Two mutations, L436(336)S in Sa.Ace1 and W516(435)R in Sa.Ace2, were found consistently associated with the insensitivity of AChE. They were thought to be the possible resistance mutations, but further work is needed to confirm this hypothesis.     

Carbon dioxide as a potential fumigant for termite control

Formosan subterranean termites, Coptotermes formosanus Shiraki, were exposed to ? 95% or 50% carbon dioxide atmospheres for intervals of 24-120 h at 26(±3)°C. A 24-h exposure to ? 95% carbon dioxide caused significant termite mortality, but 60 h were required for complete mortality. Exposure to 50% carbon dioxide for 60 h resulted in approximately 70% termite mortality, while complete mortality was recorded after 120 h. When termites were sealed in wooden blocks (90 × 90 × 152 mm), 72-96 h exposure to ? 95% carbon dioxide was necessary for complete control. A limited study with Cryptotermes brevis (Walker) suggested that this drywood termite is also susceptible to carbon dioxide fumigation, although slightly longer exposures may be required than with C. formosanus. Carbon dioxide-modified atmospheres are a viable alternative to conventional fumigants for vault fumigation of termite-infested materials, and may also be applicable to larger-scale fumigations to control structural pests.     

毒死蜱不同施药方式对黄曲条跳甲的敏感性检测

本实验以黄曲条跳甲为试虫,以毒死蜱和啶虫脒为实验药剂,分别采用浸叶法和浸虫法测定了毒死蜱和啶虫脒单剂及相关混配对黄曲条跳甲的毒力。试验结果显示,毒死蜱和啶虫脒对黄曲条跳甲的浸叶LC50值分别为53．81、268．03mg／L。啶虫脒和毒死蜱按浓度比1：4混配、1：8混配浸叶LC50值分别为58．14、41．71mg／L,浸虫法结果显示啶虫脒和毒死蜱按浓度比1：4混配、1：8混配稀释4000倍液校正死亡率分别为23．3％、26．7％。     

30%乙唑螨腈悬浮剂对荔枝叶螨不同螨态的生物活性和田间防效

为评价新型杀螨剂30%乙唑螨腈悬浮剂对荔枝叶螨的活性,对荔枝叶螨的不同螨态进行了室内活性测定及田间药效试验。室内试验结果表明:采用玻片浸渍法和浸叶法测得的30%乙唑螨腈悬浮剂对成螨的LC_(50)分别为0.977和0.248 mg/L,其毒力分别是对照药剂43%联苯肼酯悬浮剂的0.50和7.33倍;分别采用浸叶法和浸叶碟法测定了30%乙唑螨腈悬浮剂对幼若螨和卵的活性,LC_(50)分别为0.094和0.380 mg/L,活性分别为对照药剂43%联苯肼酯悬浮剂的4.07和98.40倍。处理24 h后,相同浓度的30%乙唑螨腈悬浮剂和43%联苯肼酯悬浮剂对荔枝叶螨的产卵抑制率和产卵抑制指数无明显差异,但处理48 h及72 h后,30%乙唑螨腈悬浮剂的产卵抑制率和产卵抑制指数显著高于43%联苯肼酯悬浮剂。田间防效试验结果表明:30%乙唑螨腈悬浮剂稀释4 500倍药后1 d防效达73%以上,药后20 d防效仍维持在90%以上,30%乙唑螨腈悬浮剂防治荔枝叶螨的速效性和持效性均优于43%联苯肼酯悬浮剂。     

Acetylcholinesterase point mutations putatively associated with monocrotophos resistance in the two-spotted spider mite

Molecular mechanisms of monocrotophos resistance in the two-spotted spider mite (TSSM), Tetranychus urticae Koch, were investigated. A monocrotophos-resistant strain (AD) showed ca. 3568- and 47.6-fold resistance compared to a susceptible strain (UD) and a moderately resistant strain (PyriF), respectively. No significant differences in detoxification enzyme activities, except for the cytochrome P450 monooxygenase activity, were found among the three strains. The sensitivity of acetylcholinesterase (AChE) to monocrotophos, however, was 90.6- and 41.9-fold less in AD strain compared to the UD and PyriF strains, respectively, indicating that AChE insensitivity mechanism plays a major role in monocrotophos resistance. When AChE gene (Tuace) sequences were compared, three point mutations (G228S, A391T and F439W) were identified in Tuace from the AD strain that likely contribute to the AChE insensitivity as predicted by structure analysis. Frequencies of the three mutations in field populations were predicted by quantitative sequencing (QS). Correlation analysis between the mutation frequency and actual resistance levels (LC₅₀) of nine field populations suggested that the G228S mutation plays a more crucial role in resistance (r² = 0.712) compared to the F439W mutation (r² = 0.419). When correlated together, however, the correlation coefficient was substantially enhanced (r² = 0.865), indicating that both the F439W and G228S mutations may work synergistically. The A391T mutation was homogeneously present in all field populations examined, suggesting that it may confer a basal level of resistance.     

夹竹桃叶提取物对福寿螺的杀螺活性

从夹竹桃叶中筛选杀灭福寿螺的活性成分。采用浸渍法研究了夹竹桃叶的乙醇粗提物和组分Ⅰ对不同日龄福寿螺的毒杀和抑制作用。结果表明:组分Ⅰ浸渍10、30和50日龄螺6 h的LC50值分别为2.86、5.87 和8.68 mg/L;浸渍24 h的LC50值分别为0.98、2.31和 3.16 mg/L,其中2 mg/L的组分Ⅰ浸渍10日龄螺24 h的死亡率为100%。乙醇粗提物对福寿螺的毒杀效果较差,用其100 mg/L浸渍10日龄和30日龄螺72 h的死亡率均为100%,浸渍50日龄螺120 h时,死亡率100%时的质量浓度达到200 mg/L。但乙醇粗提物和组分Ⅰ各浓度对福寿螺的活动均无抑制作用,反而有促进其上爬的作用。     

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.     

Toxicity and sublethal effects of an insecticidal soap on Aphidius colemani (Hymenoptera: Braconidae)

BACKGROUND: The effects of an insecticidal soap on the survival, fitness and behaviour of an aphid parasitoid wasp, Aphidius colemani (Viereck), were studied in the laboratory. The LC(50) (soap concentration causing 50% mortality 24 h after treatment) was determined. The survival of parasitoid larvae (% adult emergence), fitness (tibia length of adults) and number of eggs produced per female parasitoid that survived in third- and fourth-instar aphids treated with insecticidal soap LC(50) were also assessed. The LC(50) for third- and fourth-instar aphids was determined to be 3.25 g L(-1). Acceptance by female parasitoids of aphids that survived their LC(50) was also tested. RESULTS: The soap concentration causing 100% mortality in adult wasps 24 h after treatment was 17.5 g L(-1). The LC(50) was 2.75 g L(-1). Soap did not have any effect on the survival of parasitoid immatures or on the fitness or number of eggs produced per female parasitoid. Wasps that were in contact with treated aphids did not oviposit as much in them as in untreated aphids, indicating that female parasitoids detected aphids treated with insecticidal soap. CONCLUSION: These data suggest that aphid parasitoids released following treatment with insecticidal soap are likely to accept a lower proportion of the surviving aphids. Biological control programmes could be ameliorated by soap applications if the latter were made 1 day before the release of wasps in the greenhouse.     

毒死蜱微囊悬浮剂的制备及微囊化条件的优化

以脲醛树脂为壁材,采用原位聚合法制备了20％毒死蜱微囊悬浮剂,探讨了影响微囊粒径大小及分布的因素,优化了微囊化条件,并就其与毒死蜱乳油对韭菜迟眼蕈蚊Bradysia odoriphaga幼虫的毒力进行了对比。结果表明,选用苯乙烯-马来酸酐共聚乳液(SMA)为分散剂(质量分数为总体系的2.5％),乳化条件为1 500 r/min搅拌30 min,60 min内调节体系pH值至2.5,反应后期升温至45℃,在此条件下制备的毒死蜱微胶囊外形较好,平均粒径为8.9 μm,分散系数为0.182 6,包封率高达99.54％。毒力测定结果表明,乳油和微胶囊对韭菜迟眼蕈蚊幼虫的初始LC50值分别为2.05和 3.02 μg/mL;而在自然环境中放置32 d后,乳油对该虫的毒力下降较快,LC50值为100.23 μg/mL,而微囊悬浮剂毒力仍保持较高水平,LC50值为15.63 μg/mL。表明毒死蜱微囊悬浮剂兼具速效性高和持效期长的优点。     

毒死蜱在梨果实不同部位的残留及消解动态

为探明毒死蜱在梨果实不同部位的残留及消解规律,以20 a生鸭梨（Pyrus bretschneideri Rehd.cv.Yali）为试材,于果实生长期在套袋后向整株喷施48%毒死蜱乳油500倍液（有效成分960 mg/L）,分析毒死蜱向果实中的运输及分配规律;于果实成熟期在采收前向果面喷施48%毒死蜱乳油1 000倍液（有效成分480 mg/L）,分析采后毒死蜱在梨果实不同组织中的分布特征。采用乙腈萃取和GC-NPD检测方法,测定不同处理试材中的毒死蜱含量。结果表明,在果实套袋情况下,整株喷施毒死蜱后72 h内果实不同部位（果心除外）毒死蜱含量均呈现先逐渐上升而后下降趋势,其中果柄、果皮和果梗洼中毒死蜱最高含量值及其出现的时间分别为6.66 mg/kg（12 h）、2.42 mg/kg（24 h）和0.09 mg/kg（12 h）,表明套袋果实毒死蜱来源于枝叶运输,经果柄进入果实后易向果皮累积;而未套袋果施药后24和72 h果皮中毒死蜱含量分别为套袋果的12.56和7.29倍,表明套袋可有效降低果实中毒死蜱的残留量。于果实成熟期向果面喷施毒死蜱后7、14和25 d,果皮中毒死蜱残留量分别为15.54、13.70和12.81 mg/kg,占全果含量的100%,而果肉中毒死蜱残留量低于本检测方法的最低检出浓度（0.05 mg/kg）,因此果皮为果实中毒死蜱主要残留部位,且贮藏期果皮中毒死蜱不易向果肉扩散。     

甲氰菊酯对斑马鱼的毒性效应

选用模式生物斑马鱼(Brachydanio rerio)作为毒性实验动物,探索甲氰菊酯(fenpropathrin)对鱼类的毒性效应,为农业生产和环境保护提供参考数据。结果显示甲氰菊酯对斑马鱼成鱼96h LC50为3.38×10-3mg/kg,属于剧毒级,安全浓度为0.34×10-3mg/kg;较低浓度下甲氰菊酯对斑马鱼早期发育影响不显著,在较高浓度1、10mg/kg下,斑马鱼96h畸形率分别达到84%和90%,心包囊肿率分别为25%和88%,而致死效应不明显。亚致死剂量下甲氰菊酯对斑马鱼肝胰脏GPT、GOT、SOD活性有明显诱导效应,在高浓度和长时间暴露下诱导效应降低;肝胰脏中AChE活性显著地受到甲氰菊酯的抑制,抑制效应对毒物的浓度和染毒时间有良好的依赖性,是一种良好的环境污染检测生物学指标。     

药剂对小菜蛾抗性及敏感品系乙酰胆碱酯酶抑制作用比较

采用浸叶法测定了云南通海、元谋和澜沧的小菜蛾plutella xylostella田间种群对常用杀虫剂的抗药性。结果表明,云南上述地区小菜蛾田间种群对各类杀虫剂均产生了不同程度的抗性。对有机磷类药剂的抗药性为1.74~31.1倍;对菊酯类药剂的抗药性为7.41~764倍;对阿维菌素类药剂则产生了 5.60~4.06×104倍的抗性。通过离体和活体试验测定了药剂对小菜蛾头部乙酰胆碱酯酶(AChE)的抑制作用。敌敌畏和灭多威对通海抗性品系AChE离体和活体内的抑制中浓度(I50)分别是敏感品系的209、26.5倍和2.21、2.16倍;敌敌畏对通海小菜蛾种群的离体和活体内抑制中时间(IT50)小于敏感品系,分别是敏感品系的0.32和0.17倍;而灭多威对通海小菜蛾种群的离体和活体内抑制中时间(IT50)则大于敏感品系,分别是敏感品系的1.37和1.74倍。     

毒死蜱和甲氰菊酯对蚯蚓毒性与安全评价研究

按照“化学农药环境安全评价试验准则”的规定,测定了2种常用农药毒死蜱和甲氰菊酯对蚯蚓的毒性。结果表明：对蚯蚓的LC50(7d)毒死蜱为89．25mg／kg,甲氰菊酯为142．4mg／kg;LC50(14d)毒死蜱为83．63mg／kg,甲氰菊酯为36．41mg／kg。供试2种农药对蚯蚓的毒性均为低毒。