杀虫剂敌敌畏和除草剂丁草胺对斑腿树蛙蝌蚪的遗传毒性

敌敌畏和丁草胺是我国农田中使用最普遍的杀虫剂和除草剂,这些农田化学物质对当地的水生动物及种群造成很大威胁.本文以广泛分布于我国南方农田区域的斑腿树蛙蝌蚪为研究对象,用碱性单细胞电泳方法(或慧星试验)对暴露在不同浓度的敌敌畏(2.08,4.16,6.24,8.32,10.40 mg/L)和丁草胺(0.1025,0.205,0.410,0.820,1.230 mg/L)溶液中的蝌蚪进行了遗传毒性的检测.结果表明在实验室条件下,随着农药浓度的增加,蝌蚪的DNA损伤(DNA尾长与尾宽比)随之增加;敌敌畏浓度为2.08 mg/L和丁草胺浓度为0.41 mg/L时,对蝌蚪造成显著的损伤,而且农药的剂量与蝌蚪的DNA损伤(DNA尾长与尾宽比)呈显著的线性关系敌敌畏,y=1.136±0.0083x,r=0.957,P<0.01;丁草胺,y=0.968±0.0093x,r=0.964,P<0.01.本研究表明这两种农药对我国的两栖动物具有遗传毒性作用;同时也表明,在检测环境污染物对蝌蚪的基因毒性方面,碱性单细胞电泳分析是一种合适的方法[动物学报 51(3)447-454,2005].     

不同发育阶段的斑腿树蛙蝌蚪对敌敌畏遗传毒性的敏感性

敌敌畏是我国农田中使用最普遍的有机磷杀虫剂,它通过干扰对神经传导起重要作用的胆碱酯酶的活性,起到接触和摄入毒杀昆虫的作用,也对当地水生动物及种群造成很大威胁。本文以广泛分布于我国南方农田区域的斑腿树蛙蝌蚪为研究对象,用碱性单细胞电泳方法比较了暴露于不同浓度(2.08,4.16,6.24,8.32,10.40mg/L)的敌敌畏溶液中三个发育阶段的遗传毒性的差异。结果表明:敌敌畏溶液暴露24h,对早期和中期阶段的蝌蚪造成极显著的DNA损伤(P<0.01),甚至在敌敌畏浓度低达2.08mg/L的溶液中也产生显著的损伤(P<0.05);统计分析表明,DNA长宽比率与敌敌畏剂量之间呈显著的线性关系,早期阶段和中期阶段的相关系数分别为0.950(P<0.01)和0.954(P<0.01)。对于后期阶段的蝌蚪,所有浓度的敌敌畏溶液都未导致明显的DNA损伤,作者认为进入变态阶段的蝌蚪对敌敌畏的敏感性明显下降。结果同时也表明,早期和中期阶段的斑腿树蛙蝌蝌是一种监测环境遗传毒性的合适指示生物。     

Genotoxicity evaluation of the insecticide imidacloprid on circulating blood cells of Montevideo tree frog Hypsiboas pulchellus tadpoles (Anura,Hylidae) by comet and micronucleus bioassays

Acute toxicity and genotoxicity of imidacloprid (IMI) was evaluated on Hypsiboas pulchellus (Anura: Hylidae) tadpoles exposed under laboratory conditions. A lethal effect was used as the end point for lethality, whereas the frequency of micronuclei (MNs) and DNA single-strand breaks evaluated by the single cell gel electrophoresis assay were employed as end points for genotoxicity. Experiments were performed on tadpoles at stage 36 (range, 35–37) according to the classification proposed by Gosner. Mortality studies revealed an LC₅₀ (96 h) value of 84.91 mg/L IMI (95% confidence limits, 77.20–93.04). While increased frequency of MNs was observed when 15 and 30 mg/L were assayed for 48 h, only 15 mg/L increased the frequency of MNs in tadpoles exposed for 96 h. Furthermore, other nuclear abnormalities, i.e., binucleated cells and blebbed and notched nuclei, were induced in tadpoles exposed for both 48 h when treated with 15 mg/L and 96 h when treated with 15 and 30 mg/L. An increase in the genetic damage index was observed in tadpoles treated with 30 mg/L for 48 and 96 h. This study represents the first evidence of acute lethal and sublethal effects exerted by IMI on tadpoles of an amphibian species native to Argentina. Finally, our findings highlight the hazardous properties of this insecticide for nontarget living species exposed to this agrochemical.     

Acute and joint toxicity of three agrochemicals to Chinese tiger frog(Hoplobatrachus chinensis) tadpoles

We studied acute and joint toxicity of three different agrochemicals(chlorantraniliprole, flubendiamide-abamectin and penoxsulam) to Chinese tiger frog(Hoplobatrachus chinensis) tadpoles with the method of stability water tests. Results showed that the three agrochemicals increased tadpole mortality. For acute toxicity, the LC50 values after 24, 48 and 72 h of chlorantraniliprole, flubendiamide-abamectin and penoxsulam exposure were 5.37, 4.90 and 4.68 mg/L; 0.035, 0.025 and 0.021 mg/L; 1.74, 1.45 and 1.29 mg/L, respectively. The safety concentrations(SC) of chlorantraniliprole, flubendiamide-abamectin and penoxsulam to the tadpoles were 1.23, 0.30 and 0.003 mg/L, respectively. Based on these findings, chlorantraniliprole and penoxsulam were moderately toxic, while flubendiamide-abamectin was highly toxic. All pairwise joint toxicity tests showed moderate toxicity. The LC50 values after 24, 48 and 72 h of exposure were 7.08, 6.61 and 6.03 mg/L for chlorantraniliprole+penoxsulam, with corresponding values of 2.455, 2.328 and 2.183 mg/L for chlorantraniliprole+flubendiamide-abamectin, and 1.132, 1.084 and 1.050 mg/L for penoxsulam+flubendiamide-abamectin, with safe concentrations of 1.73, 0.63 and 0.30 mg/L, respectively. For toxic evaluations of pairwise combinations of the three agrochemicals, only the joint toxicity of chlorantraniliprole and flubendiamide-abamectin after 24 h was found to be synergistic, whereas all other tests were antagonistic. Our findings provide valuable information on the toxic effects of agrochemicals on amphibians and how various types of agrochemicals can be reasonably used in agricultural areas.     

L-carnitine enhances oocyte maturation and development of parthenogenetic embryos in pigs

The objective was to determine whether adding L-carnitine in IVM/IVC medium enhanced maturation and developmental competence of porcine oocytes in vitro. Oocyte maturation rates did not differ significantly among groups supplemented with 0, 0.25, 0.5, or 1 mg/mL of L-carnitine added during IVM (although 2 mg/mL of L-carnitine reduced maturation rate). Compared with control oocytes, those treated with 0.5 mg/mL of L-carnitine during IVM had greater (P < 0.05) rates of blastocyst formation after parthenogenetic activation, and these blastocysts had less (P < 0.05) apoptosis. Adding 0.5 mg/mL of L-carnitine during IVM also significantly reduced intracellular reactive oxygen species (ROS), and increased glutathione (GSH) concentrations. With or without glucose supplementation, 0.5 mg/mL of L-carnitine in the IVM medium significantly hastened nuclear maturation of oocytes. Moreover, supplementing the IVM medium with either glucose or L-carnitine increased (P < 0.05) percentages of oocytes that reached the metaphase II (MII) stage, relative to a control group. Final maturation rates in IVM medium containing either glucose or L-carnitine were not significantly different. Adding L-carnitine (0 to 2 mg/mL) to IVC medium for activated porcine oocytes did not significantly affect development. However, 0.5 mg/mL of L-carnitine in IVC medium significantly reduced reactive oxygen species levels and apoptosis in activated blastocysts, although glutathione concentrations were not significantly altered. In conclusion, adding L-carnitine during IVM/IVC improved developmental potential of porcine oocytes, and also the quality of parthenogenetic embryos, probably by accelerating nuclear maturation, and preventing oxidative damage and apoptosis.     

Effect of various heavy metals on the ATP content of spermatozoa from Arbacia lixula L.--activity of mercury]

This work reports the evaluation of some toxic effects induced by mercury on sea urchin sperm. Spermatozoa of the sea urchin Arbacia lixula L., were treated with 0.003, 0.03 and 0.06 mg/l of Hg. We estimated the amount of ATP of sperm cells with the luciferin-luciferase reaction, and we also observed motility, fertilizing activity and number of sperm bound to the eggs. The results clearly show that ATP levels are strongly affected by mercury concentrations, already three hours after the beginning of treatment. We propose the use of ATP determination in sea urchin sperm as a bioassay, because they, even more than eggs and developmental stages, readily suffer the environmental stresses.     

铅、铬胁迫对大弹涂鱼血细胞遗传损伤的彗星实验检测

通过彗星实验研究重金属Pb、Cr对大弹涂鱼的外周血细胞的影响。用不同浓度的Pb、Cr对大弹涂鱼外周血细胞进行1 h的染毒后通过单细胞凝聚电泳检测DNA损伤情况。结果表明国标浓度的Pb（0.005 mg/L）、Cr（0.05 mg/L）对大弹涂鱼外周血细胞均无明显影响;而国标10倍、100倍和1000倍浓度的Pb或Cr胁迫均会造成血细胞DNA损伤,且离子浓度与血细胞DNA的损伤程度间均存在＂剂量-效应＂,即浓度越高,DNA损伤越严重。因此大弹涂鱼血细胞可作为评价重金属遗传损伤毒性效应的敏感性生物标志物。     

葡萄糖和维生素C对镇海林蛙蝌蚪生长 及其三种酶活性的影响

营养源对动物生长发育具有重要作用。本文采用静水实验浸泡法探究葡萄糖和维生素C对镇海林蛙(Rana zhenhaiensis)蝌蚪生长及其苹果酸脱氢酶、乳酸脱氢酶及淀粉酶活性的影响。在葡萄糖实验组共分为0.5、1.0和2.0 g/L三个浓度组,维生素C实验组共分为10.0、20.0和30.0 mg/L浓度组,另设1组加曝气水作为对照组。在葡萄糖实验组中,蝌蚪存活率在不同实验组间的差异不显著(P0.05)。变态时间在0.5 g/L和1.0 g/L实验组最短,分别为(43.0±4.0)d和(43.0±3.4)d,2.0 g/L实验组最长,为(46.2±5.4)d,且实验组间差异显著(P 0.05)。变态时,0.5 g/L实验组的体重和体全长最大,且各实验组间的体重差异显著(P 0.01),但体全长差异不显著(P 0.05)。增重率在0.5 g/L实验组最高,为(9.67±1.71)mg/d,对照组最低,为(7.54±1.22)mg/d,且各实验组之间差异显著(P0.05)。维生素C实验组中,存活率在各实验组间差异不显著(P 0.05)。所有实验组蝌蚪的发育历期在实验第7、14、21和28天时均大于对照组。变态时间在所有的实验组相似,为43 d左右(P 0.05)。变态时,蝌蚪的体重(P 0.05)和体全长(P 0.05)在20.0 mg/L实验组和30.0 mg/L实验组最大,对照组最小,但各实验组间差异不显著。蝌蚪的增重率在20.0 mg/L和30.0 mg/L实验组最高,10.0 mg/L实验组最低,但不同实验组间差异不显著(P 0.05)。生化酶活性检测中,苹果酸脱氢酶(MDH)活性在葡萄糖实验组和维生素C实验组中均随实验浓度的增加而增加。乳酸脱氢酶(LDH)和淀粉酶(AMS)活性分别在葡萄糖的1.0 g/L实验组和维生素C的10.0 mg/L实验组达到最高。本研究结果表明,10.0mg/L维生素C或1.0g/L葡萄糖为镇海林蛙蝌蚪的最适外源物添加浓度,能促进蝌蚪的生长和体内酶活性。该研究结果将为镇海林蛙的养殖提供一定的理论参考数据。     

Dietary supplementation of vitamin A, C and E prevents p-dimethylaminoazobenzene induced hepatic DNA damage in rats

The preventive effect of antioxidant vitamins A, C, E and their analogues against DNA damage induced by a hepatocarcinogen p-dimethylaminoazobenzene (DAB) was assessed by comet assay. For genotoxicity (DNA damage) study, male albino rats were divided into 11 groups, consisting of four rats each. Group I served as control. Group II to VII received 1, 10, 100, 200, 300 and 400 mg per kg body wt of DAB respectively; group VIII to XI received 500 mg/kg body wt of DAB. They were sacrificed by cervical decapitation 3, 6, 12 and 24 h after treatment; livers were excised immediately and subjected to comet assay to measure DNA damage. To study the effect of vitamins, experiments were conducted on a group of 275 rats divided into 3 sets of 25 rats each. First set served as control; second set received 0.06% DAB and third set received 0.06% DAB, along with analogues of vitamins A, C and E. Rats fed with 0.06% DAB were provided water ad libitum for a period of 4 months, followed by a normal (basal) diet for further 2 months. Vitamins A (10,000-50,000 IU), C (75-1000 mg) and E (50-500 mg) and their analogues were given (per kg body wt) to the third set of rats by gavage route once in a week for a period of 6 months. The DAB induced DNA damage only at the highest tested dose of 500 mg/kg body wt. Administration of high doses of vitamin A acid, L-ascorbic acid and vit. E succinate individually prevented the DNA damage. However, administration of a mixture of these vitamins at low doses prevented the DAB-induced DNA damage, which may be due to their synergistic effect. The results indicate that there is a significant advantage in mixed vitamins therapy at low dose over the treatment with individual vitamins.     

山胡椒及其挥发物对马铃薯块茎蛾产卵选择的影响

【目的】为了确定山胡椒Lindera glauca(Sieb.Zuce)是否对马铃薯块茎蛾Phthorimaea operculella Zeller的产卵有驱避作用。【方法】利用选择性产卵试验方法在室内分别测定了山胡椒的完整果实、压碎果实、根以及不同浓度(0.25~4.0 g E/m L)的压碎山胡椒果实正己烷提取物对马铃薯块茎蛾产卵选择性的影响;在模拟仓库内测定了压碎的果实对其产卵的影响;进而在室内测定了柠檬醛、沉香醇、香叶醇和α-水芹烯不同浓度(0.00075~0.012 g/L)的溶液对其产卵选择性的影响。【结果】山胡椒完整果实、压碎果实、根在室内对马铃薯块茎蛾产卵具有驱避效果,其中压碎果实的产卵驱避效果最好,20 g压碎果实的产卵驱避率可达93.7%。山胡椒压碎果实正己烷提取物在0.5~4.0 g E/m L的浓度范围内随着浓度的升高,产卵驱避效果逐渐增强。在模拟仓库内20 g压碎的果实产卵驱避率是82.5%;0.003~0.012 g/L的柠檬醛,0.012 g/L沉香醇,0.00075~0.012 g/L的香叶醇,0.003~0.012 g/L的α-水芹烯对马铃薯块茎蛾产卵有显著的驱避效果。【结论】山胡椒及其挥发物对马铃薯块茎蛾产卵有驱避效果。     

除草剂乐草隆对红鲫的遗传毒性研究

目的　探讨除草剂乐草隆对红鲫的遗传毒性。方法　用单细胞凝胶电泳检测不同浓度的乐草隆对红鲫外周血淋巴细胞DNA的损伤作用。结果　乐草隆致毒红鲫的淋巴细胞DNA的迁移度均较阴性对照组高 (P<0 0 5 ) ,在一定浓度范围内 (0～ 7 0 0mg L)DNA损伤程度与浓度呈正相关 (r=0 982 ,P <0 0 1)。在 12h、2 4h、4 8h、96h、10d实验组DNA损伤程度均有增强的趋势。结论　乐草隆对红鲫具有一定的遗传毒性     

Benzene transformation in nitrifying batch cultures

The effect of benzene on the nitrifying activity of a sludge produced in steady-state nitrification was evaluated in batch cultures. Benzene at 10 mg/L inhibited nitrate formation by 53%, whereas at 5 mg/L there was no inhibition. For initial benzene concentrations of 0, 7, and 10 mg/L, the specific rates of NO(3)(-)-N production were 0.545 +/- 0.101, 0.306 +/- 0.024, and 0.141 +/- 0.010 g NO(3)(-)-N/g microbial protein-N.h, respectively. The specific rates of benzene consumption at 7, 12, and 20 mg/L were 0.034 +/- 0.003, 0.050 +/- 0.006, and 0.027 +/- 0.002 g/g microbial protein-N.h, respectively. Up to a concentration of 10 mg/L, benzene was first oxidized to phenol, which was later totally oxidized to acetate. Benzene at higher concentrations (20 and 30 mg/L) was converted to intermediates other than acetate, phenol, or catechol. These results suggest that this type of nitrifying consortium coupled with a denitrification system may have promising applications for complete removal of nitrogen and benzene from wastewaters.     

Short-term toxic effects of naphthalene and pyrene on the common prawn (Palaemon serratus) assessed by a multi-parameter laboratorial approach: mechanisms of toxicity and impairment of individual fitness

The short-term (96 h) toxic effects of two polycyclic aromatic hydrocarbons (PAHs), naphthalene (NAP) and pyrene (PYR), on the common prawn (Palaemon serratus) were investigated in laboratory bioassays, including a fitness related assay based on the post-exposure swimming velocity. Other effect criteria were biomarkers of neurotoxicity, oxidative stress and bioenergetics, and mortality. In the range of concentrations tested (NAP: 0.13-8 mg/L; PYR: 0.006-0.4 mg/L), both PAHs impaired the swimming velocity, induced oxidative stress and damage, and changed the activity of lactate dehydrogenase and isocitrate dehydrogenase. NAP also caused mortality (96 h-LC50=3.5 mg/L). Thus, both PAHs were able to cause toxic effects on P. serratus after a short period of exposure through the water, including the reduction of individual fitness. PYR was five folds more effective in reducing the swimming velocity of P. serratus than NAP. These findings are of interest for the marine ecological risk assessment of oil spills.     

Effects of copper sulphate concentrations during in vitro maturation of bovine oocytes

The objectives were to evaluate

1) copper (Cu) concentrations in plasma and follicular fluid (FF) from cattle ovaries; 2) the effects of supplemental Cu during in vitro maturation (IVM) on DNA damage of cumulus cells and glutathione (GSH) content in oocytes and cumulus cells; and 3) supplementary Cu during IVM on subsequent embryo development. Copper concentrations in heifer plasma (116 ± 27.1 μg/dL Cu) were similar (P > 0.05) to concentrations in FF from large (90 ± 20.4 μg/dL Cu) and small (82 ± 22.1 μg/dL Cu) ovarian follicles in these heifers. The DNA damage in cumulus cells decreased with supplemental Cu concentrations of 4 and 6 μg/mL (P < 0.01) in the IVM medium (mean ± SEM index of DNA damage was: 200.0 ± 27.6, 127.6 ± 6.0, 46.4 ± 4.8, and 51.1 ± 6.0 for supplementation with 0, 2, 4, and 6 μg/mL Cu respectively). Total GSH concentrations increased following supplementation with 4 μg/mL Cu (4.7 ± 0.4 pmol in oocytes and 0.4 ± 0.04 nmol/10⁶ cumulus cells) and 6 μg/mL Cu (5.0 ± 0.5 pmol in oocytes and 0.5 ± 0.05 nmol/10⁶ cumulus cells, P < 0.01) compared with the other classes. Cleavage rates were similar (P ≥ 0.05) when Cu was added to the IVM medium at any concentration (65.1 ± 2.0, 66.6 ± 1.6, 72.0 ± 2.1, and 70.7 ± 2.1 for Cu concentrations of 0, 2, 4, and 6 μg/mL). Percentages of matured oocytes that developed to the blastocyst stage were 18.7 ± 0.6, 26.4 ± 0.03, and 29.0 ± 1.7% for 0, 2, and 4 μg/mL Cu, and was highest (33.2 ± 1.6 %) in oocytes matured with 6 μg/mL Cu (P > 0.01). There was an increase (P > 0.05) in mean cell number per blastocyst obtained from oocytes matured with 4 and 6 μg/mL Cu relative to 0 Cu (IVM alone) and 2 μg/mL Cu. In conclusion, Cu concentrations in the FF and plasma of heifers were similar. Adding copper during oocyte maturation significantly increased both intracellular GSH content and DNA integrity of cumulus cells. Since embryo development was responsive to copper supplementation, we inferred that optimal embryo development to the blastocyst stage was partially dependent on the presence of adequate Cu concentrations during IVM.     

Tomato-oleoresin supplement prevents doxorubicin-induced cardiac myocyte oxidative DNA damage in rats

Doxorubicin (DOX) is an efficient chemotherapeutic agent used against several types of tumors; however, its use is limited due to severe cardiotoxicity. Since it is accepted that reactive oxygen species are involved in DOX-induced cardiotoxicity, antioxidant agents have been used to attenuate its side effects. To determine tomato-oleoresin protection against cardiac oxidative DNA damage induced by DOX, we distributed Wistar male rats in control (C), lycopene (L), DOX (D) and DOX+lycopene (DL) groups. They received corn oil (C, D) or tomato-oleoresin (5mg/kg body wt. day) (L, DL) by gavage for a 7-week period. They also received saline (C, L) or DOX (4mg/kg body wt.) (D, DL) intraperitoneally at the 3rd, 4th, 5th, and at 6th week. Lycopene absorption was checked by HPLC. Cardiac oxidative DNA damage was evaluated by the alkaline Comet assay using formamidopyrimidine-DNA glycosylase (FPG) and endonuclease III (endo III). Cardiomyocyte levels of SBs, SBs FPG and SBs Endo III were higher in rats from D when compared to other groups. DNA damage levels in cardiomyocytes from DL were not different when compared to C and L groups. The viability of cardiomyocytes from D or DL was lower than C or L groups (p<0.01). Lycopene levels (mean+/-S.D.nmol/kg) in saponified hearts were similar between L (47.43+/-11.78) and DL (49.85+/-16.24) groups. Our results showed: (1) lycopene absorption was confirmed by its cardiac levels; (2) DOX-induced oxidative DNA damage in cardiomyocyte; (3) tomato-oleoresin supplementation protected against cardiomyocyte oxidative DNA damage.     

汞离子和铜离子对中华大蟾蜍蝌蚪联合毒性研究

研究了重金属离子汞和铜对中华大蟾蜍（Bufo gargarizans）蝌蚪的毒性和联合毒性的影响.单种离子Hg^2＋对蝌蚪24 h、48 h的半致死浓度分别为0.712 mg/L、0.612 mg/L,Cu^2＋对蝌蚪24 h、48 h的半致死浓度分别为1.310 mg/L、0.862 mg/L,Hg^2＋和Cu2^＋共存对蝌蚪24 h及48 h半致死浓度分别为0.550-0.400 mg/L、0.379-0.300 mg/L.根据Marking的指数法求得24 h、48 h的相加指数AI分别为 0.078、0.034,表现为协同作用.     

氟对大鼠肝组织细胞凋亡与DNA损伤的影响

目的研究大鼠染氟后肝组织细胞凋亡及DNA损伤情况。方法 SD大鼠随机分为对照组、低氟组、中氟组、高氟组,每组12只,分别饮用含氟化钠为0、50、100、200 mg/L的去离子水,均饲标准营养大鼠饲料,染氟120 d。肉眼观察牙齿的变化,采用氟离子选择电极法测定大鼠尿氟,HE染色观察组织病理学变化,彗星实验检测细胞DNA损伤,流式细胞术检测肝脏组织细胞凋亡率。结果低氟组、中氟组、高氟组大鼠尿氟分别为（23.52±2.91）、（30.16±4.78）、（61.23±3.98）mg/L,均显著高于对照组（0.07±0.02）mg/L,差异有统计学意义（P〈0.01）。不同剂量染氟大鼠肝组织细胞呈现不同程度肿胀,肝组织内出现多种灶状病变。各染氟组大鼠肝细胞拖尾率及拖尾长度与相应的对照组相比,差异均有统计学意义,并且肝细胞拖尾率及拖尾长度随染氟剂量的加大而增大。不同剂量染氟组细胞凋亡率与对照组相比,均明显增高,而且高、中氟组肝细胞凋亡率显著高于低氟组（P〈0.01）。结论氟化物可导致大鼠肝细胞DNA损伤,诱导细胞凋亡,一定浓度的氟化物诱导大鼠肝细胞凋亡与DNA损伤之间存在着相关性。     

异丙甲草胺、醚菌酯和咪鲜胺锰盐对中华大蟾蜍蝌蚪的急性致死效应

采用静态换水方法研究了异丙甲草胺、醚菌酯和咪鲜胺锰盐3种农药对中华大蟾蜍(Bufo gargarizans)蝌蚪的急性毒性及其联合毒性.结果表明,在急性毒性和两两联合毒性实验中,不同浓度农药对蝌蚪的死亡率影响均显著.在24 h、48h及72 h急性毒性实验中,异丙甲草胺对中华大蟾蜍蝌蚪的半致死浓度(LC50)分别为29.81、28.81和25.83mg/L,醚菌酯为1.72、1.46和1.41 mg/L,咪鲜胺锰盐为7.43、3.75和3.22 mg/L.异丙甲草胺、醚菌酯和咪鲜胺锰盐3种农药的安全浓度(SC)分别为8.07、0.32和0.29 mg/L.异丙甲草胺为中等毒性农药,而醚菌酯和咪鲜胺锰盐均为剧毒性农药.在两两联合的毒性实验中,异丙甲草胺-醚菌酯和异丙甲草胺-咪鲜胺锰盐均表现为中等毒性,醚菌酯-咪鲜胺锰盐表现为剧毒性.在24 h、48 h及72h两两联合毒性实验中,异丙甲草胺-醚菌酯对中华大蟾蜍蝌蚪的半致死浓度(LC50)分别为18.41、15.69和13.38 mg/L,异丙甲草胺-咪鲜胺锰盐为15.56、10.45和8.11 mg/L,醚菌酯-咪鲜胺锰盐为4.17、2.84和2.00 mg/L,且其相对应的安全浓度分别为3.42、1.41和0.40 mg/L.在联合毒性评价中,异丙甲草胺-醚菌酯联合组在24 h和48 h表现为拮抗作用,醚菌酯-咪鲜胺锰盐联合组在48 h表现为加和作用,其余均表现为协同作用.同样,重复方差分析结果表明,农药浓度、染毒时间及两者相互作用对蝌蚪的存活率影响均显著.与前期发表数据进行比较,表明中华大蟾蜍蝌蚪对污染物具有很强的敏感性.这些结果将为无尾类动物的毒理学研究提供科学依据.     

Monepantel-based anthelmintic combinations to optimize parasite control in cattle

Improvement in the use of existing anthelmintics is a high priority need for the pharmaco-parasitology research field, considering the magnitude and severity of anthelmintic resistance as an important issue in livestock production. In the work described here, monepantel (MNP) was given alone or co-administered with either macrocyclic lactone (ML) or benzimidazole (BZ) anthelmintics to calves naturally infected with ML- and BZ-resistant gastrointestinal (GI) nematodes on two different commercial cattle farms. Both pharmacokinetic (PK) and efficacy assessments were performed. On Farm A, male calves (n = 15 per group) were treated with either MNP orally (2.5 mg/kg), IVM s.c. (0.2 mg/kg), ricobendazole (RBZ) s.c. (3.75 mg/kg) or remained untreated. On Farm B, eight groups (n = 15) of male calves received treatment with either: MNP, abamectin (ABM, oral, 0.2 mg/kg), RBZ (s.c., 3.75 mg/kg), albendazole (ABZ, oral, 5 mg/kg), MNP+ABM, MNP+RBZ, MNP+ABZ (all at the above-mentioned routes and doses) or remained untreated. Seven animals from each treated group (Farm B) were randomly selected to perform the PK study. MNP and its metabolite monepantel sulphone (MNPSO₂) were the main analytes recovered in plasma after HPLC analysis. The combined treatments resulted in decreased systemic exposures to MNP parent drug compared with that observed after treatment with MNP alone (P < 0.05). However, the systemic availability of the main MNP metabolite (MNPSO₂) was unaffected by co-administration with either ABM, RBZ or ABZ. Efficacies of 98% (Farm A) and 99% (Farm B) demonstrated the high efficacy of MNP given alone (P < 0.05) against GI nematodes resistant to ML and BZ in cattle. While the ML (IVM, ABM) failed to control Haemonchus spp., Cooperia spp. and Ostertagia spp., MNP achieved 99% to 100% efficacy against those nematode species on both commercial farms. However, MNP alone failed to control Oesophagostomum spp. (60% efficacy) on Farm A. The co-administered treatments MNP+ABZ and MNP+RBZ reached a 100% reduction against all GI nematode genera. In conclusion, the oral treatment with MNP should be considered to deal with resistant nematode parasites in cattle. The use of MNP in combination with BZ compounds could be a valid strategy to extend its lifespan for use in cattle as well as to reverse its poor activity against Oesophagostomum spp.     

Genotoxicity of Thermopsis turcica on Allium cepa L. roots revealed by alkaline comet and random amplified polymorphic DNA assays

This study was undertaken to evaluate genotoxic potential of Thermopsis turcica aqueous extracts on the roots of onion bulb (Allium cepa L.) by comet assay and random amplified polymorphic DNA technique. The Allium root growth inhibition test indicated that the EC₅₀ and 2×EC₅₀ values were 8 and 16 mg/ml concentrations of T. turcica aqueous extracts, respectively. The negative control (distilled water), positive control (methyl methane sulfonate, 10 mg/l) and 8 and 16 mg/ml concentrations of T. turcica extracts were introduced to the roots of onion bulbs for 24 and 96 h. The root growth, DNA damage in root cells and randomly amplified polymorphic DNA (RAPD) profiles of root tissue were used as endpoints of the genotoxicity. The comet assay clearly indicated that dose-dependent single strand DNA breaks in the root nuclei of onions were determined for the treatment concentrations of T. turcica extracts. In comparison to RAPD profile of negative control group, RAPD polymorphisms became evident as disappearance and/or appearance of RAPD bands in treated roots. The diagnostic and phenetic numerical analyses of RAPD profiles obviously indicated dose-dependent genotoxicity induced by Thermopsis extracts. In conclusion, the results clearly indicated that water extract of T. turcica has genotoxic potential on the roots of onion bulbs as shown by comet assay and RAPD technique.