Carbamate poisoning in a dairy goat herd: Clinicopathological findings and therapeutic approach

Abstract

CASE HISTORY: Approximately 1 hour after the consumption of carnations from a nearby glasshouse 55 animals from a dairy goat herd exhibited signs of possible poisoning.

CLINICAL FINDINGS: Upon clinical examination affected animals exhibited signs of salivation, tympany, tachypnoea, polydipsia, urination, diarrhoea, bradycardia, miosis, tremor and convulsions. As poisoning from an acetylcholinesterase-inhibiting insecticide was suspected, treatment with atropine sulphate was initiated at a dose of 0.3 mg/kg bodyweight. The treatment was repeated for some animals that relapsed, and was effective in all cases, with the exception of one goat kid that died.

DIAGNOSIS: Necropsy of the goat kid showed pulmonary oedema and congestion of internal organs. Toxicological analysis of stomach contents and liver of the dead animal, as well as of the carnations, revealed high concentrations of carbamates.

CLINICAL RELEVANCE: Carbamate poisoning after consumption of polluted feedstuffs or glasshouse products can be fatal for dairy goats. Atropine sulphate, at 0.3 mg/kg, can be useful in treating this condition.     

Effects of tea tree oils on removing pesticide residue in cowpea

为研究茶树油清除果蔬农药残留的效果,该试验选取豇豆为供试材料,以不同浓度的茶树油和水溶性茶树油等清洗处理,利用气相色谱和气相色谱-质谱联用检测豇豆内有机磷类、拟除虫菊酯类和氨基甲酸酯类的农药残留量,计算农药清除率。供试7种农药中,水胺硫磷、马拉硫磷、氧乐果、三唑磷、毒死蜱、氯氰菊酯和速灭威在豇豆中的初始浓度分别为：20.395、1.690、6.524、10.719、0.160、12.104和23.057 mg/kg。茶树油处理后检测结果表明,茶树油具有清除残留在豇豆中农药的能力,清除效果随茶树油浓度增加而增强;清除有机磷类农药效果较拟除虫菊酯类和氨基甲酸酯类农药明显。茶树油比去离子水、市售果蔬农残清洗剂清除农药残留效果显著,同时,相同浓度的水溶性茶树油比相应茶树油清除农药残留能力强。0.8%水溶性茶树油清除效果最佳,清除率分别为水胺硫磷80.48%,马拉硫磷94.54%,三唑磷82.79%,毒死蜱84.58%,氧乐果72.20%,氯氰菊酯80.51%,速灭威72.21%。通过研究结果可知,茶树油可作为有开发前景的果蔬清除剂。     

家蚕乙酰胆碱酯酶基因bmace在毕赤酵母GS115中的表达及其分析

【目的】构建表达家蚕乙酰胆碱酯酶（BmAChE）的毕赤酵母菌株,实现BmAChE在毕赤酵母中分泌表达并对其活性进行分析。【方法】从家蚕头部提取总RNA,经RT-PCR反转录成cDNA,通过PCR扩增出家蚕乙酰胆碱酯酶基因bmace并进行序列测定。采用PCR及重叠延伸PCR去除原bmace中的信号肽和疏水肽,并对序列中存在的2个A+T富含区进行同义密码子替换的改造。将改造后的bmace与表达载体pPIC9K连接,转化毕赤酵母GS115,甲醇诱导表达。SDS-PAGE和Western blotting鉴定重组蛋白,以Ellman法测定酶活力,毒扁豆碱和有机磷及氨基甲酸酯类农药抑制试验分析重组BmAChE（rBmAChE）的生物活性。【结果】改造后的bmace经重组毕赤酵母菌株分泌表达了1个分子量约为70 kD的蛋白,具有AChE活性,酶比活力约为1 187 U•mg-1。抑制试验显示,该rBmAChE能够被毒扁豆碱强烈抑制,并且可被皮蝇磷等10种有机磷农药和克百威等5种氨基甲酸酯类农药抑制,最低抑制浓度IC50值可达2.78×10-10 mol•L-1。【结论】成功构建分泌表达BmAChE的毕赤酵母菌株,其表达了具有良好活性的rBmAChE,可用于进一步制备有机磷及氨基甲酸酯类农药快速检测分析制剂。     

车前子乙酸乙酯部位化学成分HPLC指纹图谱研究

[目的]建立车前子乙酸乙酯部位的化学成分指纹图谱,为车前子的质量控制及品种鉴定提供依据。[方法]色谱条件为：Diamon-sil C18柱（250 mm×4.6 mm,5μm）,流动相为乙腈-0.5%的乙酸水溶液,流速为1.0 ml/min,检测波长为270 nm,采用梯度洗脱。[结果]江西道地产区10批车前子乙酸乙酯部位化学成分指纹图谱共检出12个分离度良好的共有峰。不同品种车前子乙酸乙酯部位指纹图谱相似度相差很大。[结论]该方法简便、快速,可作为车前子药材的质量控制和品种鉴定的有力手段。     

Studies on the Purification and Characterization of Soybean Esterase, and Its Sensitivity to Organophosphate and Carbamate Pesticides

Soybean esterase, a cholinesterase-like enzyme, was purified by differential centrifugation firstly, then, ammonium sulfate precipitation, dialysis, and finally, DEAE-cellulose-32 ion-exchange chromatography after extracting it from soybean seeds with phosphate buffer （0.3 mol L^-1, pH 7.0）. The extract recovery rate of the purified enzyme was 8.18% and purification fold was 91.58. The soybean esterase appeared as two bands on the denaturing SDS-PAGE with molecular weights of 24 and 37.2 kDa, respectively, which proved that it is a dimer protein consisting of two subunits. The result of nondenaturing PAGE revealed that the soybean esterase is a single band with cholinesterase-like activity using α- naphthyl acetate as the substrate and fast blue B salt as coloring agent. The esterase showed very high sensitivity to 18 kinds of organophosphate pesticides and 6 kinds of carbamate pesticides with the lowest detective limits of 0.03125- 0.0625 and 0.03125-0.25 mg kg^-1, respectively, and can meet the demands of MRL specified by the most countries.     

新型固定化乙酰胆碱酯酶传感器的制备及氨基甲酸酯农残检测

以双醛纤维素(dialdehyde cellulose,DAC)为固载酶基质材料,通过固定化酶化学键联技术制备固定化乙酰胆碱酯酶(DAC-AChE),采用自组装法制备多层壳聚糖/固定化乙酰胆碱酯酶/功能化碳纳米管生物传感器(Chi/DAC-AChE/F-CNTs/GCE),利用交流阻抗法(AC impedance,ACI),循环伏安法(cyclic voltammetry,CV)及差分脉冲伏安法(differential pulse voltammetry,DPV)考察新型传感器对农残的检测性能。结果表明:成功制备了新型固定化乙酰胆碱酯酶电化学传感器;新型传感器对氨基甲酸酯杀虫剂1-萘基-N-甲基氨基甲酸酯(carbary,CAB)具有良好的响应,检测方法响应快速,线性范围宽,灵敏度高,最低检出限可达3.5 μg·L-1,具有很好的重现性和稳定性,有关结果可为氨基甲酸酯类杀虫剂的检测提供一种新的分析方法。     

Development of ‘naked-tufted’ seed coat mutants for potential use in cotton production

Use of chemical mutagenesis has been highly successful in most major crops. The objective of this research was to develop ‘naked-tufted’ seed mutants and to incorporate this genetic trait into cotton to enhance crop quality and reduce processing costs. In 1997, six commercial cultivars were treated with 2.45% v/v ethyl methane sulfonate. In 1999, three M₃ plants were identified that had partially naked seed coats. The trait was stabilized through individual plant selections from 2000 to 2004. During 2005 and 2006, the homozygous naked-tufted M₈ mutant lines were evaluated for lint yield, lint percent, fibers/seed, fibers/mm², fiber quality, seed oil content, ginning efficiency and yarn spinning performance. Overall, the naked-tufted seed mutants had lower lint yield, lower fibers/seed, lower lint/seed, and lower fibers/mm² when compared with their original fuzzy parents. The lint turnout from the mutants was similar to the fuzzy parents and the commercial cultivars. The naked-tufted seed mutants had higher seed oil percent, 6–17% lower short fiber contents, significantly reduced seed coat neps (37–42%), higher elongation and yarn tenacity than their fuzzy counterparts. Preliminary data also showed that the naked-tufted mutants required less energy to gin.     

EMS诱变玉米花粉M_2代生物学效应研究

对EMS玉米花粉诱变的M2代进行了观察、选择和鉴定。结果表明,玉米M2苗期以叶色变异为主,其中黄化苗变异比例最大,而成株期以早熟和雄性不育变异为主。株高、穗位高等性状变异率较高。M2代果穗各性状的变异程度大都显著高于对照。     

HS-SPME-GC-MS法检测中国白酒中乙酯类成分

应用HS-SPME-GC-MS方法,检测了3种中国白酒酒样中的乙酯类成分含量,结果表明：“Shan”中含较多的乙酸乙酯、较少的乳酸乙酯和戊酸乙酯;“Si”中含较多乙酸乙酯、乳酸乙酯和己酸乙酯,是典型的浓香型白酒;“Bei”含乙酯类成分较少。