黑眶蟾蜍、黑斑蛙、中国雨蛙不同地理居群的染色体多样性

本文研究温州地区的黑眶蟾蜍、黑斑蛙、中国雨蛙的核型,分析了三个地理居群的黑眶蟾蜍、四个地理居群的黑斑蛙、三个地理居群的中国雨蛙核型.结果表明不同地理居群的同种蛙有相同的染色体数和核型模式.黑眶蟾蜍为2n=22,NF=44,核模式6+5;黑斑蛙为2n=26,NF=52,核模式5+8;中国雨蛙为2n=24,NF=48,核模式6+6.但同一种蛙的不同地理居群之间在SM数目和顺序、次缢痕或随体的位置等有所不同.说明不同地理居群的同种蛙的染色体具有丰富的多样性.     

几种化学物质对蟾蜍蝌蚪生存及生长发育的影响

在室内条件下,采用单因子急性和慢性毒性实验法,分别研究了水环境中的pH、洗涤剂、除草剂、重金属离子(Pb2 、Cu2 、Hg )对黑眶蟾蜍蝌蚪的毒性效应.结果表明,这些水体污染物不但对蝌蚪的生存造成危害,还对蝌蚪的红细胞有致畸作用;而其慢性毒害表现为蝌蚪身体畸形,肤色变浅,生长发育迟缓等.据此,可以利用黑眶蟾蜍蝌蚪对水体污染进行监测.     

中国无尾两栖类新记录种——朝鲜侧褶蛙北大核心CSCD

2021年9月在沈阳市蒲河湿地公园(41°41′54″N,122°52′19″E,海拔35 m)采集到3号无尾两栖类动物,经形态特征比较确认为侧褶蛙属(Pelophylax)金线侧褶蛙种组(P.plancyi species complex)的物种。基于线粒体12S rRNA和16S rRNA基因联合构建的侧褶蛙属部分物种最大似然系统发育树显示,采集到的侧褶蛙标本与朝鲜侧褶蛙(P.chosenicus)聚为一支,且具有较高的支持率(0.95)。综合形态和系统发育比较,确定采集到的标本为无尾目(Anura)蛙科(Ranidae)侧褶蛙属的朝鲜侧褶蛙,系中国两栖动物分布新记录种。     

虎纹蛙繁殖季节的求偶鸣叫特征(英文)

对交配季节虎纹蛙求偶鸣叫进行录制和特征分析。分析结果表明,虎纹蛙求偶鸣叫主要集中在晚上,求偶鸣叫声音主要含有3个鸣叫谐波,鸣叫主频率共有4种类型,即在第一谐波上有3个主频率段,分别为500、700和800Hz段,在第二谐波上有1个主频率段,即1800Hz段。鸣叫时程(call duration)、能环率(call dutycycle)、声强(call intensity)和鸣叫脉冲率(pulse rate)在4种主频率中变化很大。这些鸣叫参数特征的分析将对虎纹蛙生理生态学的进一步研究提供有价值的基础数据。     

聚苯乙烯暴露和清除对黑斑侧褶蛙蝌蚪身体大小和脏器系数的影响

为探究不同粒径(50 nm、200 nm和1000 nm)、浓度(清水组、低浓度组、中浓度组和高浓度组)荧光聚苯乙烯微球溶液组合对37期黑斑侧褶蛙(Pelophylax nigromaculatus)蝌蚪身体大小和脏器系数的影响,连续测定了暴露7 d时,之后清水清除饲养7 d(14 d时)和14 d(21 d时),其体重与体全长的比值(重长比)、小肠长度以及心、肝湿重的变化。重长比在200 nm粒径条件下不受聚苯乙烯微球暴露和清除影响(P>0.05);50 nm清水组14 d时低于其他浓度组(P<0.05);1000 nm中浓度组7 d时和14 d时高于21 d时(P<0.05),7 d时高浓度组低于其他浓度组(P<0.05),21 d时清水组和低浓度组都高于中浓度组(P<0.05)。小肠长度系数在200 nm高浓度组随时间而变化,7 d时至14 d时显著增加,21 d时陡降(P<0.05),1000 nm中浓度组7 d时和14 d时都显著高于21 d时(P<0.05);7 d时,1000 nm高浓度组显著低于其他浓度组(P<0.05),50 nm、200 nm均无组间差异(P>0.05)。心、肝湿重系数均不随处理时间而变化(P>0.05);心湿重系数只在50 nm 14 d时变化显著,清水组最高,高、中浓度组其次,低浓度组最低(P<0.05),肝湿重系数只在50 nm和200 nm 14 d时清水组低于其他浓度组(P<0.05)。50 nm、200 nm和1000 nm处理分别影响重长比、心和肝湿重系数,小肠长度系数和肝湿重系数,以及重长比和小肠长度系数,但与粒径和浓度均不呈线性关系。     

水位对黑斑侧褶蛙蝌蚪形态表型、白细胞数量和应激反应的影响

为探究水位变化对黑斑侧褶蛙（Pelophylaxnigromaculatus）蝌蚪形态表型、免疫功能和应激反应能力的影响,实验室内测定了水位恒定组以及水位快速、中速和慢速下降组37期蝌蚪发育至变态完成期的变态时长、身体大小和血液中各型白细胞的百分比,以及嗜中性粒细胞与淋巴细胞比值的变化。结果表明,水位快速下降显著缩短了蝌蚪的变态时长（P <0.05）,体长下降明显（P <0.05）,体重和体宽有所降低,但组间差异不显著（P> 0.05）。嗜中性粒细胞的百分比快速下降组显著高于其他处理组,单核细胞的百分比水位恒定组显著高于中速下降组（P <0.05）,嗜酸性粒细胞、嗜碱性粒细胞和淋巴细胞的百分比均无明显的组间差异（P>0.05）;嗜中性粒细胞与淋巴细胞比值快速下降组显著高于其他处理组（P <0.05）。水位加速下降导致登陆后幼蛙的个体变小,由嗜中性粒细胞和单核细胞所代表的天然免疫能力变化趋势不同,应激反应能力增强,可能不利于幼蛙的存活。     

河南新乡段卫河水对黑斑侧褶蛙早期胚胎发育的影响

研究了不同浓度新乡卫河水对黑斑侧褶蛙早期胚胎的发育毒性.结果表明:黑斑侧褶蛙胚胎在卫河水原水中不能存活;56%浓度卫河水可使胚胎畸形率和死亡率显著升高;低浓度(10%、18%、32%)卫河水能明显抑制胚胎全长的增加.在研究污染物尤其是低浓度污染物对黑斑侧褶蛙胚胎发育毒性时,生长抑制参数是比致死率和畸形率更为敏感的终点参数.卫河污染严重,存在潜在致畸变物质,应加速卫河治理.     

除草剂稻草隆对蝌蚪生长发育的影响

本文从形态学和红细胞核异常两方面观察了除草剂稻草隆对黑眶蟾蜍蝌蚪生长发育的影响. 结果表明,实验组与对照组间卵的最终出膜率差异不显著.在除草剂浓度分别为0,10,14,18,20 mg/L时,蝌蚪存活率从高到低顺序为：96.7%〉86.7%〉76.7%〉53.3%〉33.3%.经浓度为2,8,10,15 mg/L的除草剂溶液处理后,发现除草剂溶液对蝌蚪体长、体重有显著性影响（P＜0.05）,而引起的红细胞核异常率随浓度增加而升高.     

黑眶蟾蜍和黑斑蛙消化道5-羟色胺免疫活性细胞的免疫组织化学

对黑眶蟾蜍（Bufo melanostictus）和黑斑蛙（Rana nigromaculata）消化道5-羟色胺免疫活性细胞的形态、分布进行了免疫组织化学定位。5-羟色胺免疫活性细胞在黑眶蟾蜍和黑斑蛙消化道各段中均有分布,分布密度均呈升-降-升-降的波浪式分布特点,二者在幽门部和回肠都有个分布的高峰值。黑眶蟾蜍回肠最高,空肠、幽门部次之,十二指肠、直肠最低;黑斑蛙幽门部最高,回肠、空肠次之,食道、贲门部、直肠最低。5-羟色胺免疫活性细胞位于胃的胃腺上皮、食道及肠的粘膜上皮,有圆形、椭圆形、梭形、楔形等,有的有胞突。文中讨论了5-羟色胺免疫活性细胞分布型的原因及形态与功能的关系。     

同水塘分布两种无尾两栖类蝌蚪的竞争策略

实验室条件下,通过活动性水平,变态时的体重、增长率和完成变态所需时间考察同水塘分布的中华蟾蜍(Bufo gargarizans)和高原林蛙蝌蚪(Rana kukunoris)的竞争策略。实验按照2×3因子设计,即:食物资源2个水平(高、低),组合方式3个水平(10只中华蟾蜍蝌蚪,记为B组;5只中华蟾蜍蝌蚪和5只高原林蛙蝌蚪,记为BR组;10只高原林蛙蝌蚪,记为R组)。中华蟾蜍蝌蚪的活动性在食物水平低时显著低于食物水平高时,而高原林蛙蝌蚪的活动性在不同食物水平下无显著差异;食物水平低时,混合组的高原林蛙蝌蚪变态时体重和体重增长率都显著高于R组,而混合组中华蟾蜍蝌蚪与B组相比无显著差异;在不同处理组中,食物水平低时混合组中华蟾蜍蝌蚪幼体期最短。这些结果表明:中华蟾蜍蝌蚪在不同食物资源条件下所选择的生存策略可能不同,即食物资源充足时,增加活动性获取更多食物;食物资源有限时,降低活动性且提前完成变态;与中华蟾蜍蝌蚪相比,在食物资源有限时高原林蛙蝌蚪获取食物能力更强。     

Effects of hydroperiod duration on developmental plasticity in tiger frog(Hoplobatrachus chinensis) tadpoles

While developmental plasticity can facilitate evolutionary diversification of organisms,the effects of water levels as an environmental pressure on tiger frogs remains unclear.This study clarifies the relationship by studying the responses of tiger frog(Hoplobatrachus chinensis)tadpoles to simulated hydroperiods(i.e.,constant low water levels,constant high water levels,increasing water levels,decreasing water levels,rapid changes in water levels and gradual fluctuations in water levels)in a laboratory setting.ANOVA analysis showed that none of the water level treatments had any significant effect on the total length,body mass,or developmental stages of H.chinensis tadpoles half way through development(11 days old).Tadpoles raised in rapidly fluctuating water levels had protracted metamorphosis,whereas tadpoles raised under low and gradually fluctuating water levels had shortened metamorphosis.None of the water level treatments had a significant effect on the snout-vent length(SVL)or body mass of H.chinensis tadpoles at Gosner stage 42,or on the body mass of tadpoles at Gosner stage 45.However,the tadpoles raised in high levels and rapidly fluctuating water levels,significantly larger SVL at Gosner stage 45,while ones under gradually fluctuating water levels had smaller SVL than the other groups.Time to metamorphosis was positively correlated with body size(SVL)at metamorphosis in H.chinensis tadpoles.H.chinensis tadpoles under constant low water level had the highest mortality rate among all the treatments(G-test).Moreover,ANOVA and ACNOVA(with body length as the covariate)indicated that water levels had no significant effect on either the morphology(i.e.head length,head width,forelimb length,hindlimb length and body width)or the jumping ability of juvenile H.chinensis.These results suggest that the observed accelerated metamorphosis and high mortality of H.chinensis tadpoles under decreasing water level treatment was driven by density-induced physical interactions among increasing conspecifics.     

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.     

Density But Not Kinship Regulates the Growth and Developmental Traits of Chinese Tiger Frog （Hoplobatrachus chinensis） Tadpoles

Kinship and density are believed to affect important ecological processes such as intraspecific competition, predation, growth, development, cannibalism, habitat selection and mate choice, In this work, we used Chinese tiger frog Hoplobatrachus chinensis tadpoles as an experimental model to investigate the effects of kinship and density on growth and development of this species over a 73 day period. The results showed that density can affect the growth and developmental traits （survival rate, larval period, size at the limb bud protrusion/metamorphic climax and body mass at different life stages） of H. chinensis tadpoles, while kinship does not. Tadpoles took longer to develop and potential metamorphosis was greater in high density groups of both sibling and non-siblings. The interaction of kinship and density did not significantly influenced growth traits of H. chinensis tadpoles during the experimental period. For coefficient variations of each growth trait, no differences were detected between sibling and non-sibling groups. These findings provide valuable information on the basic ecology of H. chinensis which will be helpful in future studies of other anuran species.     

杀虫剂硫丹对中国虎纹蛙的毒性效应： 存活率、红细胞核异常及酶活性

农药的使用对水生动物有直接和间接的影响。本文研究了有机氯杀虫剂硫丹对中国虎纹蛙（Hoplobatrachus chinensis）蝌蚪的毒性效应。具体而言,首先检测了硫丹的安全浓度（SC）,并在急性毒性实验中评估了硫丹的毒性等级。然后检测了外周血液中红细胞核形态异常,并在慢性毒性实验中测定了酸性磷酸酶（ACP）、碱性磷酸酶（AKP）和乳酸脱氢酶（LDH）的活性。结果表明,在急性毒性中,随着硫丹溶液浓度的增加和染毒时间的延长,蝌蚪的平均死亡率显著增加,即蝌蚪存活率取决于农药剂量和染毒的时长。硫丹对中国虎纹蛙蝌蚪的96 h半致死浓度LC50值为23.38 μg/L,安全浓度为2.34 μg/L,为剧毒性农药。在血液红细胞核异常实验中,观察到5种不同类型的细胞核异常：核碎裂、双核、不等分裂、核凹陷和核空洞。血液红细胞核异常率与农药浓度呈正相关。在酶活性检测中,三种酶的活性均受硫丹浓度的影响。与对照组相比,酸性磷酸酶（ACP）和碱性磷酸酶（AKP）活性随硫丹浓度的增加而降低,乳酸脱氢酶（LDH）活性随着硫丹浓度的增加呈先升高后降低。研究结果表明,硫丹对中国虎纹蛙蝌蚪具有很高的毒性,并证明了血液红细胞核异常和特定的代谢酶可以作为环境监测的生物标志物。     

浙江地区虎纹蛙肥满度研究

分别于2007年6～10月采集了浙江金华、临安和丽水三个地区的虎纹蛙(Hoplobatrachus rugulosus)共102只进行肥满度研究,以期为我国唯一的国家二级无尾两栖类的保护提供一定的参考依据。本研究结果显示:浙江地区虎纹蛙的肥满度指标K在夏秋季节、雌雄个体以及成幼体之间均没有显著性差异(P>0.05);而重/长指标Kwl在夏秋季节和成幼体之间则存在着极显著的差异(P<0.01),而在性别间的差异性却并不显著(P>0.05)。通过对虎纹蛙样本肥满度指标K和重/长指标Kwl的比较,得知Kwl具有更高的灵敏度,并建议在无尾两栖类的肥满度研究中将两种指标结合使用,以便得到更为详尽的研究结果。     

Prey selection by the Indian tiger (Panthera tigris tigris) in Nagarjunasagar Srisailam Tiger Reserve, India

Very little is known about prey selection by the Indian tiger Panthera tigris tigris in tropical dry deciduous forests or in wild herbivore-depleted habitats with high livestock pressures. We undertook a short-term study in a large south Indian tiger reserve and examined two intensive study areas (SA's) from October 1998 – June 1999. In each area, herbivore sightings were recorded, scats were collected, and wild prey and livestock kills documented. Chital, wild boar and sambar were the most abundant among the wild herbivores in the study area. Scat analysis revealed wild boar (Sus scrofa) being the most common prey followed by chital (Axis axis) and sambar (Rusa unicolor). Livestock comprised less than 7% of diet intake. Here, the tigers consumed a lower mean prey mass (56.3 kg) than in other reserves. Our study suggests that in tropical dry deciduous forests with low natural prey density, smaller prey species, and high livestock biomass, tigers preferentially kill smaller prey and generally avoid livestock predation.     

Evolutionary significance of oral morphology in the carnivorous tadpoles of tiger frogs, genus Hoplobatrachus (Ranidae)

We describe the larval stages of three species of the Asian-African tiger frogs Hoplobatrachus chinensis, H. occipitalis and H. tigerinus . The tadpoles of all three species are very similar, with peculiar oral features: (1) double rows of needle-like labial teeth, (2) strong emarginations on the large jaw sheaths and (3) keratinized spurs on the buccal floor. Characters 1 and 2 (and perhaps 3) are probably related to the carnivorous habits of these tadpoles. A molecular phylogeny based on 2430 base pairs of two nuclear and four mitochondrial genes corroborated monophyly of Asian and African Hoplobatrachus, and identified Euphlyctis as their sister group. Tadpoles of the latter genus lack buccal spurs and double labial tooth rows but share large jaw sheaths, the upper with a medial projection. Therefore, the common ancestor of Euphlyctis and Hoplobatrachus probably was also characterized by this state, and may have been facultatively carnivorous. Further carnivorous specializations in Hoplobatrachus could explain why tiger frogs have been so successful in populating arid environments where ponds are at high risk of desiccation. Larval morphology may prove to be the key innovation which enabled them to disperse, in the Late Cenozoic, into their current very wide distribution area in Asia and Africa.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 81 , 171–181.     

低温胁迫下虎纹蛙的生存力及免疫和抗氧化能力

全球气候变化是造成世界范围内两栖类种群衰退和灭绝的重要因素之一。随着极端天气出现变得日趋频繁,非季节性的、短期且剧烈的气温变化可能会严重干扰两栖类动物的生存与种群稳定。监测了浙江省金华市南山野生虎纹蛙(Hoplobatrachus rugulosus)分布区冬季的环境气温,并参考监测数据在实验室条件下研究了虎纹蛙在短期梯度降温(2℃/24 h)和急性冷暴露(即冷休克)(2℃)下的生存力及冷休克对机体免疫功能和抗氧化能力的影响。结果表明,虎纹蛙在冬季(2009-12-01—2010-03-31)经历的温度范围普遍在0—14℃之间,主要遭遇的低温区间在0—4℃,主要高温区间在10—14℃。通过梯度降温实验,发现温度降至12℃累积死亡率约为28.1%,10℃为87.5%,8℃为100%。在一定温度范围内,虎纹蛙死亡率与环境温度呈显著负相关(Pearson test,r=-0.952,P<0.05)。经曲线拟合,回归方程计算可得半数致死温度为11.5℃。虎纹蛙在冷休克处理下,在第6 h累积死亡率为45%,12 h为80%,24 h达到100%。虎纹蛙死亡率与冷休克时间呈显著正相关(Pearson test,r=0.91,P<0.05),半数致死时间为7.6 h。此外,冷休克(2℃,6 h)显著抑制了虎纹蛙脾脏巨噬细胞呼吸爆发强度(t=3.827,df=6,P<0.05)、全血吞噬活性(t=5.388,df=3.037,P<0.05)及胃溶菌酶活力(t=6.37,df=6,P<0.05);肝脏(t=0.773,df=8,P>0.05)和肾脏(t=0.164,df=4.542,P>0.05)组织脂质过氧化物产物丙二醛(MDA)的含量虽无明显变化,但肝脏(t=-2.817,df=6,P<0.05)和肾脏(t=-11.302,df=6,P<0.05)组织抗氧化物谷胱甘肽(GSH)含量及肝脏(t=-3.3,df=6,P<0.05)超氧化物歧化酶(SOD)活性均显著升高。上述结果表明虎纹蛙对低温较为敏感,冷休克能够诱导机体的免疫抑制,并导致机体对抗氧化物质需求的增加。可以推测,当遭遇极端低温天气时,低温胁迫可能会严重干扰虎纹蛙生理机能,加大种群的生存压力和疾病感染的风险。     

Oviposition habitat selection by the mosquito, Culiseta longiareolata: effects of conspecifics, food and green toad tadpoles

Abstract. 1. Ovipositing females should choose egg-laying sites which maximize the growth and survivorship of their offspring. Accordingly, they should avoid habitats with high densities of competitors and predators and should choose sites with adequate food. We tested experimentally the oviposition response of the mosquito, Culiseta longiareolata Macquart, to extra food and two species which are both potential competitors and predators of its offspring: conspecific larvae and green toad ( Bufo viridis ) tadpoles.
2. We conducted the experiment in outdoor artificial pools where wild C. longiareolata females were presented with choices of all combinations of the presence or absence of: (1) C.longiareolata larvae, (2) B.viridis tadpoles, and (3) extra food (a mixture of ground fish food and mouse chow).
3. Oviposition rates averaged approximately 4 times greater in response to extra food.
4. The presence of the tadpoles, particularly later in the experiment when they were larger, caused a significant reduction (approximately 50%) in oviposition rate.
5. We did not detect a statistically significant effect of conspecific larvae on oviposition habitat selection.
6. Our results indicate that Culiseta females choose habitats for oviposition based on food availability. They also tend to avoid habitats containing Bufo tadpoles, presumably to avoid predation and/or competition.