Toxic Effects of Three Heavy Metallic Ions on Rana zhenhaiensis Tadpoles

Heavy metal pollution is widespread in some areas of China and results in contamination of land, water, and air with which all living organisms interact. In this study, we used three heavy metallic ions （Cu^2＋, Pb^2＋ and Zn^2＋） to assess their toxicity effects on mortality, blood biomarker and growth traits （body length and body mass） of Rana zhenhaiensis tadpoles. The results showed that the toxicity levels of the three metallic ions were different when conducted with different experiment designs. For acute toxicity tests, Cu^2＋ was the most toxic with the highest tadpole mortality. The mortalities of tadpoles showed significant differences among the treatments at the same exposure time endpoints （24, 48, 72 and 96h）. Results from repeated measures ANOVA indicated that metallic ion concentration, exposure time and their interactions significantly affected the mortalities of R. zhenhaiensis tadpoles. Also, the toxicity effects of all binary combinations of the three metallic ion treatments showed synergism. The half lethal concentrations （LCs0） decreased with increasing exposure time during the experimental period, and the safe concentration （SC）values of Cu^2＋, Pb^2＋ and Zn^2＋ were different from each other. Combined and compared LC50 values with previous data reported, it is suggestes that the toxicity levels of metal pollution to anuran tadpoles should be species-and age-related. For blood biomarker tests, Zn^2＋ was the most toxic with the highest total frequencies of abnormal erythrocytic nucleus. All three metallic ions caused higher abnormal erythrocytic nucleus compared with control groups. In a chronic toxicity test, Pb^2＋ was the most toxic with lowest growth traits. Survival rate （except for 18 days）, total body length and body mass showed significant differences among the treatments. These findings indicated that tadpoles of R. zhenhaiensis should be as a bioindicator of heavy metals pollution.

Genotoxicity of the Pesticide Dichlorvos and Herbicide Butachlor on Rana zhenhaiensis Tadpoles

Genotoxicity of dichlorvos and butachlor on erythrocytes of Rana zhenhaiensis tadpoles was investigated by the alkaline single-cell gel electrophoresis assay or comet assay.Tadpoles were treated for 24h in the laboratory with different concentrations of the testing agents,2.256,4.512,6.768,9.024,11.280mg/L for dichlorvos and 0.292,0.584,0.876,1.168,1.460mg/L for butachlor,to use the comet Assay to test for the significance of dosage responsiveness to an increase in DNA damage,asmeasured by themean DNA tail length-to-width ratio.The concentrations of 4.512mg/L dichlorvos and 0.876mg/L butachlor resulted in highly significant increases in DNA damage of the tadpoles.There were linear correlations between themean DNA tail length-to-width ratio and the concentrations of the two test substances.Our results showed that the two commonly used agricultural chemicals caused dose dependent DNA damage of amphibians,and that comet assaymight be a useful tool formeasuring DNA damage of tadpoles exposed in the field.

饲料β-胡萝卜素水平对镇海林蛙蝌蚪生长发育的影响

为探讨饲料中添加不同剂量的β-胡萝卜素对于镇海林蛙(Rana zhenhaiensis)蝌蚪生长、幼蛙形态和运动表现及肝脏酶活的影响。将镇海林蛙蝌蚪(Gosner 25~26)288尾随机分为4个处理组。各组分别添加0、50、500和1200 mg/kgβ-胡萝卜素,直至镇海林蛙蝌蚪变态发育完全。结果显示:(1)不同处理组间蝌蚪发育中期的体重、体长和尾长组间差异不显著,而蝌蚪中期发育的历期在组间差异显著,对照组中期发育的历期显著快于β-胡萝卜素处理组。(2)不同处理组间蝌蚪的增重率、特定生长率和变态率差异不显著,对照组的变态时间显著短于β-胡萝卜素处理组,β-胡萝卜素处理组则随着浓度的增加而变态时间缩短。(3)不同处理组间幼蛙体长、头长、前臂及手长和后肢长差异显著,但幼蛙体重差异不显著。(4)不同处理组间幼蛙平均跳跃距离差异显著,50和500 mg/kgβ-胡萝卜素处理组幼蛙平均跳跃距离显著大于另外两组。(5)幼蛙的过氧化氢酶(CAT)活力在不同处理组间差异显著,50 mg/kgβ-胡萝卜素组的酶活显著大于其它处理组,但是幼蛙肝脏总超氧化物歧化酶(T-SOD)活力差异不显著。综上所述,饲料中添加500 mg/kgβ-胡萝卜素试验组,可以显著减缓镇海林蛙蝌蚪的早期发育,延长变态时间,形成较大的幼蛙个体,积累能量较多,运动性能显著增强,个体的适合度增加。