PVA-SbQ法固定乙酰胆碱酯酶检测农药残留

以PVA-SbQ固定乙酰胆碱酯酶,检测蔬菜中的农药残留.对六种农药的提取效率在50%-105%之间.低于20%的丙酮和低于30%的甲醇对固定乙酰胆碱酯酶的活性具有一定的促进作用.六种农药在四种蔬菜中的检出限在0.05-0.8mg·kg-1之间.     

4种浮萍提取物的抗菌活性和黄酮含量

浮萍(Lemnaceae)生长在微生物丰富的污水表面,为了研究浮萍抗菌机制,为浮萍抗菌物质活性筛选以及动植物病原菌和人类致病微生物感染机制研究提供依据,用不同极性的溶剂对浮萍进行提取,牛津杯法(Oxford Cup)及最小抑菌浓度(MIC)测定比较绿萍zh0018、少根紫萍zh0224以及多根紫萍zh0003、zh0225四种浮萍提取物的体外抗菌活性,并用分光光度法结合高效液相色谱(HPLC)测定分析提取物总黄酮含量以及成分差异.结果表明:浮萍甲醇提取物、95%乙醇提取物对枯草芽孢杆菌、金黄色葡萄球菌、铜绿假单胞菌、大肠杆菌、农杆菌均有不同程度的抗菌活性,其中多根紫萍zh0225甲醇提取物抗菌活性最强(MIC:6.25 mg/mL),其次为绿萍;各萃取物中乙酸乙酯相活性最强(MIC:0.78 mg/mL),其次为正丁醇相.分光光度法测得绿萍甲醇提取物总黄酮含量较高,为提取物的9.8%,其次为少根紫萍甲醇提取物,为8.1%,其中多根紫萍zh0225乙酸乙酯相含量最高,为萃取物的22.46%,其次为正丁醇相,为17.2%.HPLC测得不同浮萍提取物黄酮成分差异较大,两株多根紫萍成分相似而含量不同.研究表明,结合浮萍自身的特性和提取物抗菌活性的大小及黄酮含量,筛选多根紫萍zh0225乙酸乙酯萃取相作为优良浮萍品种提取活性物质以及开展药物生产具有巨大潜力.     

朱砂叶螨乙酰胆碱酯酶的原核表达及其活性分析

乙酰胆碱酯酶(ACh E)在神经信号传导过程中起重要作用,是氨基甲酸酯类和有机磷类农药的主要作用靶标.对朱砂叶螨ACh E进行体外表达和纯化,并分析其活性.将朱砂叶螨ace基因序列插入到原核高效表达载体p ET-30a中,构建表达载体p ET-30a/ace,转入表达菌株Escherichia coli BL21(DE3)中,IPTG诱导高效表达;镍柱纯化目的蛋白,经SDS-PAGE和Western Blot鉴定目的蛋白;以纯化的螨ACh E为抗原制备抗螨ACh E特异性抗体;改良的Ellman法测定螨ACh E活性.结果获得了相对分子质量(Mr)约为68×103的较高纯度的螨ACh E蛋白,检测大部分为可溶性表达,酶活检测具有乙酰胆碱酯酶活性,并制备了107效价的抗螨ACh E特异性抗体.采用重组ACh E(IC50=5.4 mmol/L)检测毒扁豆碱抑制活性比螨粗酶液(IC50=58.7 mmol/L)灵敏度提高11倍.本研究构建了螨ACh E体外高效表达载体并获得了具有较好活性的重组螨ACh E,可为抗ACh E杀螨剂的体外高通量筛选和以ACh E为靶标的农药残留检测奠定基础.     

太阳鱼脑乙酰胆碱酯酶的分离纯化

采用硫酸铵沉淀–普鲁卡因胺亲和柱层析方法,对太阳鱼(Lepomis gibbosus)脑部乙酰胆碱酯酶分离纯化,纯化倍数为234倍,回收率为25.67%,经SDS-PAGE电泳检测,呈现单一的条带,相对分子质量约为68 000,表明其纯度达到电泳纯.     

基于杀螨剂靶标朱砂叶螨乙酰胆碱酯酶体外表达的新型杀螨活性化合物筛选

乙酰胆碱酯酶(Acetylcholinesterase,AChE)是保证神经信号在生物体内正常传递的关键酶,是有机磷和氨基甲酸酯类农药的作用靶标.本研究在前期朱砂叶螨乙酰胆碱酯酶基因原核表达及最适反应条件摸索的基础上,大规模表达纯化朱砂叶螨AChE,进一步研究其酶学性质.研究结果显示,在温度为25℃、其他条件最适的情况下,测定酶K_m值为0.67 mmol/L,K_(cat)/K_m值为13.53 mmol L~(-1) s~(-1).随后,从SPECS化学库筛选到了55个候选AChE抑制性化合物,获得了9个抑制活性优于或相当于毒扁豆碱的朱砂叶螨AChE抑制化合物,并且它们对朱砂叶螨有较好的毒性.本研究为乙酰胆碱酯酶抑制剂的筛选以及以乙酰胆碱酯酶为靶标的农药开发提供了理论依据和实验基础.     

3种有机磷农药对水生生物的乙酰胆碱酯酶抑制效应的物种敏感度分析初探

通过筛选敌敌畏、马拉硫磷和对硫磷3种有机磷农药对水生生物的急性毒性数据和乙酰胆碱酯酶抑制效应数据,构建物种敏感度分布曲线进行了比较分析。结果表明,敌敌畏对水生生物的急性毒性和乙酰胆碱酯酶抑制效应的大小顺序为:酶体内抑制效应酶体外抑制效应急性毒性;马拉硫磷和对硫磷的乙酰胆碱酯酶抑制效应数据不足但趋势相似,顺序为:酶体内抑制效应急性毒性酶体外抑制效应。敌敌畏的急性毒性和酶体外抑制效应的5%危害浓度(HC5)分别为2.07μg·L~(-1)和1.53μg·L~(-1),两者相差1.4倍。在水质基准推导中,乙酰胆碱酯酶抑制效应数据对有机磷农药的水生生物基准具有重要的参考价值。     

核桃粕中抗氧化和糖苷酶抑制活性成分

为了发掘核桃粕潜在的利用价值,提取核桃粕的化学成分,测定其抗氧化活性和糖苷酶抑制活性并进行活性跟踪.结果显示,95%乙醇提取物具有一定的生物活性,抗氧化能力用DPPH法测定自由基清除率来表示,测得IC_(50)为1.482 8 mg/m L,抑制糖苷酶的能力用α-淀粉酶和α-葡萄糖苷酶的抑制活性表示,测得IC_(50)分别为6.679 8 mg/m L和0.3572 mg/m L.为了得到活性物质的最大提取效率,分别选用95%乙醇、纯水、70%乙醇、60%丙酮4种溶剂提取样品,并从提取得率、抗氧化活性、糖苷酶抑制活性等方面进行评估,最后选用70%乙醇为最佳提取溶剂.对70%乙醇粗提物进一步分相萃取,结合柱色谱、薄层色谱结晶等分离方式,从乙酸乙酯相中得到化合物Y-10-1,其清除自由基IC_(50)为0.0372 mg/m L,抑制α-淀粉酶和α-葡萄糖苷酶IC_(50)分别为0.017 9 mg/m L和0.617 3 mg/m L.抗氧化活性和糖苷酶抑制活性分别以维生素C和阿卡波糖作为阳性对照,测得阳性对照的抗氧化IC_(50)为0.100 2 mg/m L,α-淀粉酶抑制IC_(50)为1.462 mg/m L,α-葡萄糖苷酶抑制IC_(50)为0.165 1 mg/m L.经比较,Y-10-1的抗氧化活性和α-淀粉酶抑制活性均优于阳性对照.经液相和质谱鉴定,基本确定Y-10-1为没食子酸.本研究说明Y-10-1具有潜在的使用和开发价值.     

杀虫剂敌敌畏对黑眶蟾蜍蝌蚪的毒性影响

采用静态换水法研究了杀虫剂敌敌畏（dichlorvos，DDVP）对黑眶蟾蜍（Bufo melanostictus）蝌蚪的急性毒性和生长发育、生理活动的影响．结果表明，DDVP对黑眶蟾蜍蝌蚪的96h半数致死浓度（LC50，96h）为51．61mg／L．暴露于2．24mg／L的DDVP溶液中10d和20d，蝌蚪的体长日增长率分别降低24．9％和9．5％，并引起黑眶蟾蜍蝌蚪酯酶同工酶（EST）酶带数和强弱发生改变，抑制了蝌蚪的酯酶同工酶活性，而超氧化物歧化酶（SOD）比活受到的影响相对较小．暴露于6．72mg／L的DDVP溶液中10d和20d，蝌蚪的体长日增长率分别降低27．0％和38．8％，酯酶同工酶（EST）酶带的强弱发生明显的改变，SOD酶比活明显低于对照组．说明杀虫剂敌敌畏对黑眶蟾蜍蝌蚪可造成一定的毒性影响．图2表4参22     

鼠尾藻提取物对亚历山大藻的化感效应

研究不同浓度的鼠尾藻组织蒸馏水及4种不同极性有机溶剂(甲醇、丙酮、乙醚、氯仿)提取物对亚历山大藻的生长抑制作用.结果表明:鼠尾藻组织甲醇提取物对亚历山大藻的生长抑制活性最强,大于3×10-8 mg kg-1浓度作用下亚历山大藻在1 d内完全死亡;鼠尾藻组织蒸馏水抽提物与对照组相比在各个浓度时均能显著抑制亚历山大藻的生长,但即使在最高浓度1.6×10-6mg kg-1下亦对其无致死效应;其他3种有机溶剂提取物对亚历山大藻的生长无明显影响.表明鼠尾藻组织中含有的对亚历山大藻具有抑制作用的活性物质具有较高的极性.对鼠尾藻甲醇提取物进行液液萃取,将其分离为石油醚相、乙酸乙酯相、正丁醇相和蒸馏水相,并对亚历山大藻进行生物活性检测.结果发现,石油醚相和乙酸乙酯相具有较强的杀藻活性,表明脂肪酸可能是鼠尾藻组织内抑制亚历山大藻生长的克生物质的重要组成成分之一.图3参36     

五氯硝基苯对小鼠脑AChE的影响及多器官氧化损伤研究

测定五氯硝基苯(PCNB)对小鼠脑乙酰胆碱酯酶(AChE)的影响及多器官的氧化损伤作用.对昆明种小鼠采用灌胃法进行200,400和600 mg·kg-1三个剂量染毒,每日一次,分别于染毒4,8,12和16d测定AChE活性、过氧化氢酶(CAT)活性和脂质过氧化(LPO)产物丙二醛(MDA)的含量.结果表明:(1)小鼠脑AChE活性受到抑制,并表现为明显的时间和剂量依赖性;(2)肝、肾CAT活性变化显著,脂质过氧化作用明显;(3)高剂量毒物在染毒16d时对脾脏脂质过氧化作用明显增强;(4)脑和血清CAT活性随染毒时间延长先升高后降低,脂质过氧化作用不明显.PCNB对小鼠可能具有神经毒性,对肝、肾脏的氧化损伤作用较强.     

Larvicidal potential of different solvent extracts and oleanolic acid derivative from Coccinia indica against vector mosquitoes

The mosquito larvicidal activities of different solvent extracts (chloroform, ethyl acetate, and methanol) from the leaves of Coccinia indica were studied. Among the extracts tested, the methanol extract showed the highest larvicidal activity against early fourth instar larvae of Culex quinquefasciatus (LC₅₀?=?142?mg?L^?1), Aedes aegypti (LC₅₀?=?158?mg?L^?1) and Anopheles stephensi (LC₅₀?=?161?mg?L^?1) after 24?h of exposure period. The lowest larval mortality was observed with the ethyl acetate extract with the LC₅₀ values of 927, 872, and 730?mg?L^?1 against Cx. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The chloroform extract showed the moderate larvicidal activity against the mosquito species tested with the LC₅₀ values from 446 to 546?mg?L^?1. A new triterpene was isolated from the methanol extract using bio-assay guided separation and identified as an oleanolic acid derivative by NMR, IR and mass spectral studies. The compound showed prominent larvicidal activity against Cx. quinquefasciatus, Ae. aegypti and An. stephensi with the LC₅₀ values of 5.6, 5.0, and 4.8?mg?L^?1, respectively. The results of the present investigation offer an opportunity for developing alternatives to rather expensive and environmentally hazardous organic insecticides.     

Larvicidal,repellent, and histopathologic effects of Citrullus colocynthis against the malaria vector

Abstract

This research is conducted to determine the larvicidal, repellent and pathological effects of fruit extracts of C. colocynthis on malaria vector mosquito, Anopheles stephensi Liston, 1901. The fruit water and ethanol extract of C. colocynthis were concentrated for larvicidal and repellent activities on human volunteer against An. stephensi. The temephos larvicide was tested as the positive control. A concentration of 300?mg/L had full-scale mortality and the most toxic effect. The LC₅₀ and LC₉₀ values of C. colocynthis water and ethanol extracts were 180and 283?mg/L, respectively. The LC₅₀ and LC₉₀ values of temephos were 0.2706 and 1.6932?mg/L, respectively. The ED₅₀ and ED₉₀ values of the plant extract were 0.2 and 1.3?mg/cm², respectively. Thin sections from treated larvae exhibited gross histopathologic effects of C. colocynthis fruit extracts on gut epithelial cell layer by vacuolization of foregut cuboidal and midgut columnar cells. This study suggested that the fruit extracts of C. colocynthis had significant larvicidal and repellent activity to be used as an ideal approach to control the malaria vector mosquito.     

Inhibitory effect of the plant Boerhavia diffusa l. against the dermatophytic fungus Microsporum fulvum

Antifungal activity (reduction in colony diameter) of various extracts (pt. ether, chloroform, ethyl acetate, ethyl alcohol and aqueous) of aerial and root parts of Boerhavia diffusa (Nictaginaceae) was screened against dermatophytic fungi Microsporum fulvum. Statistically significant increase has been recorded in the % inhibition of the target fungal species with increasing test concentrations (1000-5000 ppm) of chloroform, ethyl acetate and ethyl alcohol extracts of the root. The maximum % inhibition observed in various solvent extracts of root was about 26% (chloroform), 46% (ethyl alcohol) and 57% (ethyl acetate) at 5000 ppm concentration with time exposure of 10 days. The colony diameter of the target mycelial colony decreased with increasing supplementation of the phytoextract, showing the presence of significant amount of some antifungal phytochemical moiety.     

Extraction,isolation, and identification of various environment friendly components from cock's comb (Celosia argentea) leaves for allelopathic potential

In this study, the aqueous extract of leaves of the plant cock's comb (Celosia argentea) was fractioned into five batches using n-hexane, dichloromethane, ethyl acetate, and n-butanol, respectively. High performance liquid chromatography analysis of the methanol re-dissolved residues of 3 polar fractions, including dichloromethane, ethyl acetate, and n-butanol separately which exhibited altogether 12 compounds identified as 4-hydroxycinnamic acid (p-coumaric acid), 4-hydroxybenzoic acid, protocatechuic acid, caffeic acid, m-coumaric acid, gallic acid, 4-hydroxybenzaldehyde, m-hydroxybenzaldehyde, 2,4-dihydroxybenzoic acid, pyrogallol, 3,5-dihydroxybenzoic acid, and genetic acid was categorized by retention time using known compounds. Similarly, the mass confirmation of most of the identified compounds was achieved by injecting 20 µl of composite sample of these three polar fractions into liquid chromatography–mass spectrometry. Similarly, the physiochemical analysis of C. argentea aqueous extract showed the presence of various constituents such as phenols, flavonoids, tannins, and glycosides. All four organic fractions were examined for phytotoxicity against Lepidium sativum. Analysis of variance showed significant differences among these fractions against germination and growth of L. sativum. Further allelopathic effects appeared to be concentration dependent. Thus, C. argentea may prove effective as a substitute of herbicides to control the weeds like L. sativum from attacking crops and to protect the environment from the health hazard effects of herbicides.     

Evaluation of the antimutagenic and antiradical potentials of extracts from the tubers of (Tunisian) Cyperus rotundus

The mutagenic potential of aqueous, total oligomers flavonoids (TOF), ethyl acetate and methanol extracts from tubers of Cyperus rotundus L. was assessed using Ames Salmonella tester strains TA98 and TA100, and SOS chromotest strain Escherichia coli PQ37 with and without metabolic activation (S9). None of the different extracts produced a mutagenic effect. Likewise, the antimutagenicity of the same extracts was tested using the “Ames test” and the “SOS chromotest”. Our results showed that C. rotundus extracts possess antimutagenic effects against S. typhimurium TA98 and TA100 strains towards the mutagen aflatoxin B1 (AFB1), similar to E. coli PQ37 strain against AFB1 and Nifuroxazide mutagens using the SOS chromotest assay. A free radical scavenging test was used in order to explore the antioxidant capacity of the extracts obtained from the tubers of C. rotundus. TOF, ethyl acetate and methanol extracts showed an important free radical scavenging activity towards the 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical. These extracts showed an IC50 value of respectively 5, 20 and 65?µg?mL^?1. The beneficial effects of TOF, ethyl acetate and methanol extracts of C. rotundus were assessed by antioxidant and antimutagenic activities.     

Toxicological study of plant extracts on termite and laboratory animals

Toxic activity of leaf extracts of Polygonum hydropiper L. and Pogostemon parviflorus Benth. were tested in the laboratory against tea termite, Odontotermes assamensis Holm. Both the tested extracts caused mortality of the termite. The highest toxic activity (100%) was found in the 2.0% chloroform extracts of P. hydropiper. The chloroform extract of P. hydropiper was explored for possible mammalian toxicological effects. The LD50 was 758.58 mg/kg in male albino mice. Subcutaneous injection of sub-lethal dose of extract into male mice once a week for 6 weeks failed to express any significant influence on WBC, RBC count and blood cholesterol.     

Clove bud oil: an efficient, economical and widely available oil for the inhibition of wheat seed germination

Pre-harvest sprouting refers to the precocious germination of the grain in the spike prior to harvest as a result of moist weather conditions at harvest time. From the agricultural viewpoint, it is necessary to impose an exogenous dormancy to wheat seeds in order to improve the resistance of seed to pre-harvest sprouting. In this regard, we found that clove bud essential oil is a strong inhibitor for wheat seed germination. The extract obtained from clove bud by supercritical fluid extraction using CO₂ as solvent minimized the number of extracts to two compounds, eugenol and eugenyl acetate. Eugenol, as the main constituent of the oil, was responsible for its strong inhibitory activity in wheat seeds. The aqueous solution of clove bud oil was submitted to germination assay at various concentrations from 50 to 400 mg/L. Complete inhibition of seed germination was recorded when the concentration was 400 mg/L. Roots and sprouts have similar sensitivity to inhibitory effect. In an empirical study, the synergistic cooperation of eugenol and eugenyl acetate from clove bud oil in the inhibition of seed germination was found to be a 1:1 ratio. The clove bud essential oil is widely available and will broaden the horizon of applications for natural and safe inhibitors in the fields.     

Antibacterial,antiviral and antioxidant activities of aerial part extracts of Peganum harmala L. grown in Tunisia

Ethyl acetate, chloroform, butanol, and methanol extracts from the aerial part of Peganum harmala were tested for antibacterial, antioxidant, and antiviral activities. The antibacterial activity was evaluated by the determination of minimum inhibitory concentration (MIC) using the solid medium technique. Oxacillin, amoxicillin, ticarcillin, cefotaxim, and amphotericin were used as control agents. The antiviral activity was evaluated against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and Coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. The antioxidant activity was evaluated using two tests: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging and ammonium thiocyanate methods. Among tested extracts; the chloroform extract displayed a higher antibacterial activity against Gram-positive than Gram-negative bacteria. The butanol extract demonstrated the highest antioxidant activity. The methanol extract showed significant antiviral activity against CoxB-3 virus. The chloroform extract may be an important source of bactericidal compounds against Gram-positive bacteria.     

Toxicological and larvicidal activities of Alzanzalakhet,Melia azedarach against cucurbit fly,Dacus ciliatus at Hail Province in Saudi Arabia

Alzanzalakhet, Melia azedarach L. (Sapindales: Meliaceae) is a common medicinal plant which is found in Hail desert in Saudi Arabia and traditionally used for various purposes. In this study, toxin effectiveness of the ethanolic, acetonic, and aqueous extracts of M. azedarach leaves was tested to determine a larvicidal activity against the cucurbit fly, Dacus ciliatus (Loew) (Diptera: Tephritidae). Different treatments with M. azedarach extracts varied significantly in their impact on mortality of D. ciliatus larvae. Results showed that all tested extracts in the three solvents exerted a positive pattern for reducing larvae of D. ciliatus. However, the ethanolic M. azedarach extract was the most potent compared with other tested solvents. The highest mortality of the D. ciliatus larvae was shown to be 94%, 74%, and 68% when the concentration was 1?×?10⁵?ppm for ethanolic, acetonic, and aqueous extracts, respectively. In contrast, the concentration 1?×?10¹?ppm resulted in the lowest mortality of D. ciliatus larvae at 40%, 34%, and 12% for ethanolic, acetonic, and aqueous extracts, respectively. Overall, the results of this study suggest that the ethanolic extract of M. azedarach leaves may be used as a sustainable method for controlling D. ciliatus larvae.     

Phytoconstituents evaluation and antihyperglycemic and antihyperlipidemic effects of Mahonia leschenaultia Takeda in streptozotocin-induced diabetic rats

Hyperlipidemia is an associated complication of diabetes mellitus. Many spices and herbs are known to be hypoglycemic. Mahonia leschenaultia Takeda (Berberidaceae) (MLT) is a traditional medicinal plant that is commonly used against diseases in India and other Asian countries. The aim of this study is to identify the phytoconstituents by gas chromatography–mass spectrography (GC–MS) and evaluate the antihyperglycemic and antihyperlipidemic effects of the ethanol extract of MLT on streptozotocin (STZ)-induced diabetic rats. Phytoconstituents like oleicacid, 9,12-octadecadienoic acid (Z, Z)-n-hexadecanoic acid were identified as high % peak when compared with other compounds by GC–MS. STZ-induced diabetic animals were fed with plant extracts at different doses (250 or 500 mg kg^?1 body wt) of ethanol extract of MLT. The results of the study revealed a significant increase in total hemoglobin and glycosylated hemoglobin along with the reduction in hyperglycemia and hyperlipidemia effects in MLT-treated STZ-induced diabetic rats compared to diabetic only rats. Further, ethanol extracts of MLT-treated animals at 500 mg kg^?1 showed that shrinkage of β cells of islets of Langerhans was restored as evidenced by histological studies of pancreas of diabetic animals. Thus the ethanol extract of MLT might serve as a reliable adjuvant for antihyperglycemic and antihyperlipidemic effects of STZ, and may be promising for development of phytomedicines for diabetes mellitus.