中国林蛙蝌蚪对盐度和pH的耐受性

分别在正常盐度不同 pH值、正常 pH值不同盐度和相同水温 18 3～ 2 0 7℃的条件下 ,进行中国林蛙蝌蚪对盐度和 pH值的耐受性实验。结果表明 ,在pH值为 6 8～ 7 0时 ,盐度在 2 0～ 3 0g·L-1对蝌蚪 96h存活率没有明显影响 ;随着盐度的增加 ,蝌蚪存活率下降 ;盐度对蝌蚪 2 4、4 8、72和 96h的TLm 值分别为 8 79,7 12 ,5 6 4和 3 83g·L-1,安全浓度为 1 73g·L-1。在盐度为 0 18g·L-1时 ,pH值在 4 3～ 9 7范围内对蝌蚪 96h存活率没有明显影响 ;pH值对蝌蚪的 2 4 ,4 8,72和 96h的TLm 值分别为 10 33,10 18,10 0 8和 10 0 2。生产上中国林蛙蝌蚪生存的适宜 pH值为 6 0～ 9 0 ,适宜盐度为 1 0～ 2 0g·L-1。     

壬基酚对中国林蛙蝌蚪生长发育的毒性效应

为评价壬基酚对中国林蛙(Rana chensinensis)蝌蚪的急性毒性,将26～28期的中国林蛙蝌蚪暴露于浓度为0.05～0.5 mg·L-1壬基酚(NP)的水体中进行急性毒性试验.结果表明:24、48、72、96h蝌蚪的死亡几率与浓度对数的回归方程分别为y=8.4087χ 10.202、y=9.5104χ 11.745、y=10.284χ 12.498、y=10.619,χ 13.095;半数致死浓度(LC50)分别为0.24、0.20、0.19、0.17 mg·L-1;安全浓度(SC)为0.017mg·L-1;96h的LC0为0.14 mg·L-1.证明林蛙蝌蚪的死亡几率与一定范围的NP浓度呈线性正相关.为探讨低浓度壬基酚对中国林蛙蝌蚪生长发育的影响,将26期林蛙蝌蚪分别在100、60、30、10μg·L-1 NP的水体中连续暴露直至完全变态,并以3、0.3 μg·L-1雌二醇(E2)作为阳性对照,分别统计林蛙蝌蚪暴露于NP和E2,中20、40d和完成变态后幼蛙的体质量、全长以及从26期到完全变态所需的时间.结果表明,NP浓度在100 μg·L-1以下,对林蛙蝌蚪不能致死.100、10 μg·L-1NP与3 μg·L-1E2的效应相似,均可滞后蝌蚪变态时间,降低蝌蚪变态后幼蛙的体质量.说明NP浓度在100μg·L-1以下不直接损伤蝌蚪机体,但可以通过干扰内分泌活动影响林蛙幼体的生长发育.     

中国林蛙的热生物学研究进展

本文对中国林蛙卵、胚胎、蝌蚪、幼体和成体等不同发育阶段的热生物学研究进行了综述,温度对中国林蛙不同发育阶段均有不同的影响。11～13℃是卵孵化的最适温度,卵孵化率接近100%,胚胎在7～22℃温度范围内均能正常发育;10～20℃是蝌蚪的最佳生长发育温度,成活率都在95%以上;5℃下蝌蚪不能完成变态,在15～30℃之间,温度越高雄性比例越大,15～25℃是成体生活的最适温度。这些研究结果可为中国林蛙的人工养殖提供科学依据。     

中国林蛙变态蝌蚪对pH、盐度和碱度的适应性

在水温16～18℃的野外条件下,采用单因子急性毒性实验法,研究了水环境中pH、盐度和碱度对中国林蛙(Rana chensinensis)变态蝌蚪的毒性效应．结果表明,中国林蛙变态蝌蚪对pH的适应范围为4．3～9．7,最低耐受限3．6,最高耐受限10．7;对盐度的最高耐受限为9．98g·L^-1,适应盐度上限7．14g·L^-1,安全盐度上限1．70g·L^-1;对碱度的最高耐受限为19．96mmol·L^-1,适应碱度上限8．76mmol·L^-1。安全碱度上限2．41mmol·L^-1．野外变态蝌蚪饲养池水体pH应控制在6．5～8．5,盐度控制在2．0g·L^-1以下,碱度不超过4．0mmol·L^-1．中国林蛙变态蝌蚪是一种狭酸碱、低耐盐、低耐碱生物。     

温度对中国林蛙卵孵化和孵出热耐受性的影响

为了解中国林蛙(Rana chensinensis)卵的孵化率、发育起点温度、发育有效积温、孵化后蝌蚪的成活率和蝌蚪的热耐受性,将当天产出的中国林蛙卵采集回实验室,分别置于5个不同温度(10℃、15℃、20℃、25℃和30℃)下孵化,观察和测量记录卵的孵化率、孵化3d后蝌蚪的成活率、全长及每个温度下卵的发育历期。利用温度梯度装置观察记录孵化10d后蝌蚪的最适温度、逃避温度和致死温度。光照周期设为14L∶10D,湿度设为85%。结果表明,温度对中国林蛙卵的孵化率影响显著;孵化3d后蝌蚪的成活率随着温度的升高而降低,30℃下孵化的蝌蚪在2d之内全部死亡;温度对中国林蛙卵孵化3d后蝌蚪全长的影响极显著,蝌蚪的全长随着温度的升高而增加;应用直线回归法和直接最优化法计算中国林蛙卵的发育起点温度和有效积温,分别为0.51℃、65.29日度和0.38℃、65.52日度,直接最优化法优于直线回归法。经过4个不同温度(10℃、15℃、20℃和25℃)孵化10d后蝌蚪的最适温度分别为(13.2±1.6)℃、(15.7±1.9)℃、(17.3±2.1)℃和(19.5±2.3)℃;逃避温度分别为(28.8±1.4)℃、(30.2±1.6)℃、(31.6±1.3)℃和(33.1±1.8)℃;致死温度分别为(32.9±1.7)℃、(33.8±1.5)℃、(35.2±1.2)℃和(36.7±0.9)℃。经过不同温度孵化10d后蝌蚪的最适温度、逃避温度和致死温度都存在显著差异。     

农药乐果对昭觉林蛙蝌蚪的耐受性研究

采用水生生物急性毒性试验方法,研究乐果对昭觉林蛙Rana chaochiaoensis蝌蚪的急性毒性影响,同时运用扫描取样法对正式实验的昭觉林蛙蝌蚪进行行为观察.结果 表明:昭觉林蛙蝌蚪在试验组中正常静止行为、异常静止行为、正常运动行为、异常运动行为分别占74.08%、9.41%、0.58%、2.36%;在对照组中,只有正常静止行为和正常运动行为,分别占90.99% 和9.01%.乐果对昭觉林蛙蝌蚪24 h、48 h、72 h、96 h的半致死浓度 (LC50) 分别为:76.97 mg/L、67.85 mg/L、56.61 mg/L、52.80 mg/L,呈现出线性关系,安全浓度 (SC) 为5.28 mg/L.因此,乐果对昭觉林蛙蝌蚪当属于低毒性农药.但是当乐果残留量超过安全浓度时,将会影响昭觉林蛙蝌蚪的正常生长发育,并对其行为产生影响.     

四指马鲅(Eleutheronema tetradactylum)受精卵和仔鱼对不同盐度的耐受性

本文观察了11个盐度梯度(5、7、9、11、13、15、20、25、28、30、35、40、45,28作为对照组)下四指马鲅(Eleutheronema tetradactylum)不同发育阶段的受精卵的沉浮性质和孵化情况以及初孵仔鱼在不投饵条件下的存活情况,并测定不投饵存活系数(survival activity index,SAI)。结果表明:第一次分裂前和原肠期两发育阶段的受精卵在盐度低于25的海水中均呈沉性,盐度为25时则悬浮在水层中部,盐度28及以上呈浮性;第一次分裂前的受精卵对盐度改变较敏感,而原肠期受精卵对盐度胁迫产生一定的耐受性;盐度的改变对初孵仔鱼的存活率和SAI值影响比较大。综合分析孵化率、仔鱼畸形率以及SAI值,推测四指马鲅受精卵孵化的适宜盐度范围为25~35,最适孵化盐度为35。     

直链烷基苯磺酸钠对中国林蛙抗菌肽基因表达的影响

近年来由于生境丧失及环境污染等原因,中国林蛙野生种群急剧减少.为深入研究水体中普遍存在的阴离子表面活性剂直链烷基苯磺酸钠(linear alkylbenzene sulfonate,LAS)对中国林蛙生存的影响,通过急毒实验确定LAS对分别处于发育阶段26、32、42、46期的中国林蛙的半致死浓度.同时利用1、4、8 mg/L LAS处理不同发育阶段中国林蛙,研究LAS对中国林蛙存活率、发育状态的影响,并利用real-time PCR技术检测中国林蛙抗菌肽基因表达水平变化.结果表明,当处理时间为96 h,LAS浓度达到12.33 mg/L以上时会对不同发育阶段中国林蛙产生较为严重的致死效应.1~8 mg/L浓度范围的LAS虽然对中国林蛙存活和发育没有明显影响,但是改变了其抗菌肽基因表达模式.特别是中国林蛙胚后发育后期(32期)和变态期(42期),4~8 mg/L LAS会在不同程度上抑制prepropalustrin-2CE3、preprobrevinin-2CE1和preprotemporin-1CEd1抗菌肽基因表达.总体而言,高浓度LAS对中国林蛙免疫系统造成一定影响,可能使该物种更易受到致病菌的侵袭.研究结果为化学污染物对中国林蛙免疫系统影响积累了实验数据,同时为研究中国林蛙抗菌肽基因表达的调控通路提供一定的理论依据.     

密度对中国林蛙蝌蚪生长发育的影响

将中国林蛙（Rana chensinensis）蝌蚪分别饲养在40只／L、80只／L、120只／L、160只／L、200只／L和240只／L6个不同的密度条件下,通过定时定量喂食及测量其全长、尾长、体宽、体重和死亡率的变化,分析密度对中国林蛙蝌蚪生长发育的影响。实验结果表明：（1）饲养密度和生长发育时间及两者的交互作用对中国林蛙蝌蚪的生长发育影响都极其显著;（2）在40只／L的密度下,中国林蛙蝌蚪的全长、尾长、体宽、体重的增长速率最快,在240只／L的密度下,中国林蛙蝌蚪的全长、尾长、体宽、体重的增长速率最慢;（3）在相同时间内,40只／L的密度条件下中国林蛙蝌蚪的存活率最高,在240只／L的密度条件下中国林蛙蝌蚪的存活率最低;（4）中国林蛙蝌蚪的生长发育速度随饲养密度的升高而减慢。     

广西棱皮树蛙Theloderma kwangsiensis生物学初步研究

2010年至2012年对海南鹦哥岭两栖类动物进行了调查,野外采集到广西棱皮树蛙Theloderma kwangsiensis的蛙卵,对蛙卵进行了人工孵化,对蝌蚪与幼蛙进行了人工饲养。结果显示:该蛙繁殖时间为3月中旬,蛙卵孵化时间为14 d,6个月左右完成变态,蝌蚪唇齿式为Ⅰ∶2+2/1+1∶Ⅱ。该蛙种群数量极其稀少,建议加强保护。     

Sublethal Effects of Nitrite on Eastern Tiger Salamander (<Emphasis Type="Italic">Ambystoma tigrinum tigrinum</Emphasis>) and Wood Frog (<Emphasis Type="Italic">Rana sylvatica</Emphasis>) Embryos and Larvae: Implications for Field Populations

Ephemeral pools, which can have high animal biomass and low dissolved oxygen, may be prone to nitrite accumulation. As such, it is important to understand how exposure to nitrite might affect development and growth of amphibians that breed in these ephemeral pools. Wood frog (Rana sylvatica) and eastern tiger salamander (Ambystoma tigrinum tigrinum) embryos and tadpoles and young larvae were exposed to elevated concentrations of nitrite derived from sodium nitrite: 0, 0.3, 0.6, 1.2, 2.1, 4.6, and 6.1 mg l⁻¹ NO₂–N. Increasing nitrite exposure slowed embryonic and larval development in both the eastern tiger salamander and the wood frog, reduced growth in tiger salamander embryos and larvae, and delayed metamorphosis in the wood frog. At concentrations less than 2 mg l⁻¹ NO₂–N nitrite delayed hatching, and at concentrations above 2 mg l⁻¹ time to hatching decreased causing more individuals to hatch at less developed stages. Nitrite also increased asynchrony in tiger salamander hatching. The sublethal effects of nitrite on amphibian development, growth and hatching could have serious repercussions on amphibian fitness in ephemeral environments. Potential increases in mortality on field populations caused by sublethal effects of nitrite are discussed.     

Effects of prochloraz and ethinylestradiol on sexual development in Rana temporaria

A wide range of environmental xenobiotics that mimic hormones (endocrine-disrupting chemicals) may cause alterations in sexual development or reproductive function in aquatic organisms such as amphibians when exposed during early sensitive stages. We exposed tadpoles of the Common frog, Rana temporaria, from hatch to metamorphosis, to two different endocrine disruptors, the synthetic estrogen 17 alpha-ethinylestradiol and the fungicide prochloraz. The object of the study was to assess the effects of these two compounds on the sexual development of the tadpoles by investigating sex ratio, gonadal development, sex steroid concentrations and vitellogenin induction. Histology revealed that a large percentage of all groups were juvenile hermaphrodites at metamorphosis. Tadpoles exposed to 115 and 251 microg/L prochloraz showed a significant increased proportion of males. However, the testosterone concentrations were depressed in those groups. Ethinylestradiol in concentrations of 77 and 159 ng/L EE(2) increased whole-body calcium levels in a dose-dependent manner indicating induction of the egg yolk protein vitellogenin, verified also by gel electrophoresis. The study shows that ethinylestradiol may induce vitellogenesis and prochloraz may affect the sexual development in Common frogs.     

双酚A对中国林蛙生精细胞凋亡和Bax、Bcl-2表达的影响

为探讨双酚A(BPA)对两栖动物生精细胞凋亡及相关蛋白Bax和Bcl-2表达的影响.将雄性中国林蛙(Rana chensinensis)分别暴露于10-7、10-6、10-5 mol/L BPA水体中持续3 d、5 d、7 d,取其精巢组织.用原位末端转移酶法(TUNEL)和甲基绿-派诺宁法(Methyl Green-Pyronine)检测生精细胞凋亡,用免疫组织化学方法检测生精细胞的Bax和Bcl-2表达.结果显示,各BPA处理组中国林蛙生精细胞凋亡指数(Apoptotic index,AI)均显著高于对照组,10-6 mol/L与10-7 mol/L BPA处理组生精细胞的AI差异不显著,10-5 mol/L BPA处理组生精细胞的AI与前两组相比显著增高;在同一BPA浓度处理组,生精细胞AI随处理时间的延长而增高.与对照组相比,各BPA处理组Bax表达上调,Bcl-2表达下调,差异均显著;生精细胞AI与Bax/Bcl-2表达呈正相关.这些结果提示,BPA可导致中国林蛙的生精细胞凋亡,而生精细胞凋亡的发生与Bax/Bcl-2表达比值的变化密切相关.     

Direct influence of melatonin on the thyroid and comparison with prolactin

Melatonin administered in vivo had previously been shown to inhibit thyroid cell proliferation and subsequent in vitro thyroxine (T(4)) secretion in anuran tadpoles. Melatonin in vitro also directly reduced the sensitivity of the thyroid to thyrotropin (TSH). The present work sought to determine whether melatonin directly affected baseline, unstimulated T(4) secretion, and to compare its effect with that of prolactin (PRL). Thyroids from larval Rana catesbeiana or adult Rana pipiens were incubated in control or melatonin (0.01 to 100 microg/ml) media. Melatonin directly inhibited T(4) secretion by thyroids from both tadpoles and frogs at all concentrations of melatonin used and at both prometamorphic and climax tadpole stages. PRL, used in vitro at 10 microg/ml, did not influence the response of the thyroid to TSH (0.2 microg/ml) in young tadpoles, or the baseline secretion of T(4) by thyroids at any stage of larval life except climax, when T(4) secretion was significantly decreased by the third day of culture. Thus although both melatonin and PRL have been shown to antagonize the action of T(4) in vitro, and to decrease metamorphic rate, melatonin is a much more effective thyroid gland inhibitor than PRL.     

金线蛙早期胚胎发育的初步观察

报道了金线蛙的早期胚胎发育。自受精卵期至鳃盖完成期共分为 2 6个时期 ,其发育历程及各时期胚胎外形特征与黑斑蛙基本相似 ,但孵化期比黑斑蛙推迟三个胚期而早于泽蛙 ,在水温 ( 2 0± 0 5)℃及常温 ( 2 1 5～ 2 4℃ )条件下 ,其胚胎发育的全时程分别为 2 1 2 94小时和 1 70 95小时。     

中国林蛙蝌蚪的食性

本文对河北产中国林蛙蝌蚪的食性进行了系列的试验分析,是对其人工养殖不同发育阶段的饵料配方进行科学探索的一部分。结果表明：不同种类的单一饵料对蝌蚪的生长发育产生不同影响;蝌蚪不同发育时期对饵料趋向性不同;一定比例的人工合成饵料适宜蝌蚪期饲喂。     

Influence of Melatonin on the Rate of Rana pipiens Tadpole Metamorphosis In Vivo and Regression of Thyroxine-Treated Tail Tips In Vitro

Metamorphosis of Rana pipiens tadpoles may be retarded when the light phase of the light/dark (LD) cycle is shortened or when thyroxine (T₄) is given in the dark because melatonin peaks during the dark. Injection of premetamorphic tadpoles in spontaneous metamorphosis with melatonin (15 μg) retarded tail growth and hindlimb development on 18L:6D but had no significant effect on 6L:18D. During induced metamorphosis (30 μg/liter T₄), melatonin injections retarded tail resorption on 18L:6D and accelerated it on 6L:18D, but did not affect the hindlimb. When melatonin was injected during T₄ immersion at different times in the photophase on 18L:6D (L onset 0800 hr), tail regression was retarded by melatonin at 1430 or 2030 hr. At 0830 hr, shrinkage of tail length was accelerated whereas tail height was not affected. Tail tips in vitro induced to resorb by 0.2 μg/ml T₄ in Niu-Twitty solution regressed more slowly in the presence of melatonin (10 or 15 μg/ml) than with T₄ alone on both 6L:18D and 18L:6D. The findings implicate melatonin in LD cycle effects on tadpole metamorphic rate in vivo , show the importance of the time of melatonin injections, and indicate that melatonin antagonizes the metamorphic action of T₄ at the tissue level.     

Postembryonic development of the olfactory and vomeronasal organs in the frog Rana chensinensis

We studied by light microscopy the histological development of the olfactory and vomeronasal organ in tadpoles of the Chinese forest frog, Rana chensinensis, from postembryonic periods to the end of metamorphosis. Unlike Bufo americanus, the olfactory epithelium in larval R. chensinensis is not divided into dorsal and ventral branches in the rostral and mid-nasal regions. The olfactory epithelium in the dorsal portion of the buccal cavity in larval R. chensinensis may correspond to the ventral olfactory epithelium of Bufo, which has been argued to provide a chemosensory function in the tadpoles analogous to the role of taste buds in adults. Bowman's glands were present in the olfactory epithelium of R. chensinensis only after the appearance of the forelimbs during metamorphosis. The appearance of Bowman's glands in the olfactory epithelium at this time suggests that the nose first begins to detect odorants in the air, and this is thus also a metamorphic event. The vomeronasal epithelium appeared a little earlier than the vomeronasal gland in R. chensinensis, unlike in toads (bufonids). This study supports Eisthen's hypothesis that the most recent common ancestor to the tetrapods was aquatic and once had a vomeronasal organ, and that this has been lost in various evolutionary lineages.