氟离子慢性暴露对中国林蛙蝌蚪生长发育的毒性效应研究

氟元素以不同的结合形式广泛存在于自然界中,适量氟的摄取有益于龋齿预防和骨骼发育,然而过量氟的摄取会对动物及人体健康造成危害。近年来,大量的人类活动导致水环境中氟含量持续升高。为探究水域氟污染对中国林蛙的毒性影响,本研究以中国林蛙(Rana chensinensis)胚胎为试验材料,对卵黄栓期(G12期)胚胎进行了0、0.7、4.2、19.4、42.8 mg·L~(-1)F-慢性水体暴露直至胚胎发育到变态高峰期(G42期)的研究。分别于暴露25 d和40 d后取样测定了蝌蚪全长、体长、体重和发育分期;此外,分析了F-慢性暴露对变态率、G42期蝌蚪的全长、体长、体重和后肢长以及G42期蝌蚪骨骼发育的影响。结果表明:暴露25 d时,4.2 mg·L~(-1)F-处理组促进了林蛙蝌蚪的生长发育,而42.8 mg·L~(-1)F-处理组显著抑制了蝌蚪的生长发育;暴露40 d时,19.4 mg·L~(-1)F-和42.8 mg·L~(-1)F-处理组蝌蚪的生长发育均受到显著抑制。持续进行慢性暴露78 d后,4.2 mg·L~(-1)F-处理组蝌蚪的变态率显著升高,而42.8 mg·L~(-1)F-处理组蝌蚪的变态率受到了显著抑制。此外,42.8 mg·L~(-1)F-处理组G42期蝌蚪形态指标(全长、体长和后肢长)以及骨骼发育均受到抑制。依据G42期中国林蛙蝌蚪的生长发育指标和变态率为观察指标,氟离子慢性暴露对中国林蛙蝌蚪的最低可观察效应浓度(LOEC)为0.7 mg·L~(-1)。研究表明,水环境中高浓度的氟污染会对中国林蛙蝌蚪的生长发育、变态和骨骼发育等造成潜在的不利影响,水体氟污染的生态毒性效应理应引起高度重视。     

Re-evaluation of thyroid hormone signaling antagonism of tetrabromobisphenol A for validating the T3-induced Xenopus metamorphosis assay

We developed the T3-induced Xenopus metamorphosis assay, which is supposed to be able to sensitively detect thyroid hormone(TH) signaling disruption of chemicals. The present study aimed to validate the T3-induced Xenopus metamorphosis assay by re-evaluating the TH signaling antagonism of tetrabromobisphenol A(TBBPA), a known TH signaling disruptor. According to the assay we developed, Xenopus tadpoles at stage 52 were exposed to 10–500 nmol/L TBBPA in the presence of 1 nmol/L T3. After 96 hr of exposure, TBBPA in the range of 10–500 nmol/L was found to significantly inhibit T3-induced morphological changes of Xenopus tadpoles in a concentration-dependent manner in term of body weight and four morphological endpoints including head area(HA), mouth width(MW), unilateral brain width/brain length(ULBW/BL), and hind-limb length/snout-vent length(HLL/SVL).The results show that these endpoints we developed are sensitive for characterizing the antagonistic effects of TBBPA on T3-induced metamorphosis. Following a 24-hr exposure,we found that TBBPA antagonized expression of T3-induced TH-response genes in the tail,which is consistent with previous findings in the intestine. We propose that the tail can be used as an alternative tissue to the intestine for examining molecular endpoints for evaluating TH signaling disruption. In conclusion, our results demonstrate that the T3-induced Xenopus metamorphosis assay we developed is an ideal in vivo assay for detecting TH signaling disruption.     

氨氮对中华绒螯蟹仔蟹肝组织RNA/DNA及肝细胞结构的影响

以中华绒螯蟹（Eriocheir sinesis）仔蟹为研究对象。采用静水试验法,温度在20℃左右,用氨氮与过滤海水分设5个质量浓度组（2.04、6.35、9.04、11.75和19.13 mg.L-1）,同时以过滤海水为对照组（氨氮质量浓度为0.30 mg.L-1）,每一个质量浓度组设3个平行样本,对中华绒螯蟹仔蟹进行6 d胁迫试验。每2 d观察仔蟹的蜕壳情况,并分别于实验后的2、4和6 d取仔蟹肝组织样,应用激光共聚焦技术分析测定RNA/DNA荧光相素比,同时用透射电镜观察6 d后,最高浓度组和对照组仔蟹肝细胞结构的变化情况。试验结果表明;对于氨氮的胁迫,仔蟹的变态率随染毒浓度的增高而降低,同时变态蜕壳的时间也会产生延迟现象。氨氮的胁迫会引起仔蟹的肝组织细胞内的DNA和RNA的质量分数逐渐降低,导致RNA/DNA比率的不断下降。在高质量浓度氨氮的胁迫下,使得仔蟹肝细胞线粒体部分解体、胞质空泡化和染色质浓缩、转运泡数量增多,体积增大,细胞出现许多空泡和微绒毛消失等一系列的损伤变化。破坏了仔蟹自身机能正常的代谢和生长水平。     

Effects of combinations of malathion and cypermethrin on survivability and time of metamorphosis of tadpoles of Indian cricket frog (Fejervarya limnocharis)

This study was undertaken to determine the effect of environmentally realistic concentrations of two commonly used pesticides viz., malathion and cypermethrin, using a fully 3 × 3 factorial experiments on the survivability and time of metamorphosis in a common rice paddy field frog (cricket frog) Fejervarya limnocharis under laboratory conditions. The results suggest that cypermethrin is more toxic than malathion and combinations of higher concentrations of cypermethrin (50 μg/L) with malathion (250 and 500 μg/L) are more deleterious to the survivability of tadpoles. With increasing cypermethrin concentration, the survivability of tadpole decreased (r = ?0.986, P = 0.108). But cypermethrin alone induced early metamorphosis among the surviving tadpoles. However, there was a delay in the time required for metamorphosis induced by malathion and its combination with cypermethrin. The delay in metamorphosis may indicate the altered physiological fitness of the individual. The emergent froglets will be subjected to environmental stressors like high temperature and less humidity of post-monsoon tropical climate that could enhance negative influence triggered by pesticides.     

全氟辛磺酸（PFOS）对非洲爪蟾（Xenopus laevis）生长发育、甲状腺和性腺组织学的影响

为了揭示全氟辛磺酸(PFOS)的甲状腺激素和性激素干扰效应,并探索运用爪蟾动物模型同时检测多种环境内分泌干扰效应的可能性,将NF48阶段非洲爪蟾(Xenopus laevis)蝌蚪暴露于0.01、0.1和1mg·L-1PFOS中6个月,检测PFOS对爪蟾生长、变态、甲状腺和性腺的影响.结果表明,在各取样时间,PFOS组爪蟾体长、体重和蝌蚪尾长与对照组均无显著差异(p>0.05);2个月后,PFOS组比对照组平均慢1个发育阶段,4和6个月后,0.01mg·L-1PFOS组反而比对照组分别快1和2个发育阶段.6个月后,PFOS组甲状腺出现滤泡上皮细胞增生、胶质减少甚至空泡化等现象,且随着PFOS浓度的增加而加重;各组幼蛙性腺出现间性、睾丸珍珠状和先天性萎缩等畸形现象,外观为间性的性腺主要表现为睾丸组织中产生类似雌性个体的卵巢腔.对照组幼蛙的雌雄比为0.5:1,而暴露组分别为2.3:1(0.01mg·L-1组)、4.5:1(0.1mg·L-1组)和5:1(1mg·L-1组).上述结果表明,PFOS对爪蟾的变态过程具有小剂量刺激效应,能引起甲状腺组织结构的损伤,导致睾丸组织的雌性化和雌雄性比的异常升高,表现出明显的甲状腺激素和性激素干扰效应,可以认定为一种环境内分泌干扰物.实验同时表明非洲爪蟾可以用于多种环境内分泌干扰效应的同时检测.     

Marsh Frog (Rana ridibunda Pall.) in Cooling Ponds in the Middle Urals

Environmental conditions in cooling ponds of thermal power plants are favorable for marsh frogs. Their populations inhabiting Verkhne-Tagil and Reftinskoe reservoirs differ in some traits, such as size and age composition, growth rate after metamorphosis, spawning type, fecundity, and characteristics of larval development. Differentiation in body size manifests itself at the early stages of terrestrial life and progresses at later stages. As a consequence, larger individuals may reach maturity and participate in reproduction at an earlier age.     

Chronic exposure effects of copper on growth,metamorphosis and thyroid gland,liver health in Chinese toad,Bufo gargarizans tadpoles

The impact of copper exposure on Chinese toad (Bufo gargarizans) tadpoles was investigated in this study. First, the 96h LC50 value of copper was 8.697 μM, by means of a 4 d acute toxicity test. Second, we studied the chronic effects of copper on B. gargarizans tadpoles at control, 0.025, 0.1 and 1.0 μM concentration. Survival, body length, body weight, developmental stage, incidence of metamorphic climax, and size at metamorphic climax were determined. In tadpoles developed to metamorphic climax (stage G42), liver and thyroid gland were assessed histologically. Copper at 0.1 and 1.0 μM could inhibit tadpole growth and prolong tadpole metamorphic progress relative to controls. Tadpole size (total length and weight) at stage G42 is also affected in the 0.1 and 1.0 μM treatments. In addition, histological examinations have revealed that 1.0 μM copper could cause significant pathological changes and hepatocytes degeneration in liver. Furthermore, histomorphological measurements indicated that copper at 0.1 and 1.0 μM reduced thyroid gland size, diameter and number of follicle. In conclusion, our study suggests that Cu could damage the liver and thyroid gland, so growth and metamorphosis of B. gargarizans tadpoles were inhibited resulted of disrupting liver metabolism and THs homeostasis.     

Morphological and behavioral alterations induced by endocrine disrupters in amphibian tadpoles

Developmental toxicity effects of endocrine disrupter chemicals, acephate and cypermethrin were studied in Bufo melanostictus tadpoles. Thirty developing eggs of B. melanostictus were exposed to each concentration (0.01, 0.05, 0.1, 0.5, and 1?µg?L^?1) of acephate or cypermethrin in the laboratory (temperature: 23?±?1°C; photoperiod: 11.5–12.5?h). Eggs maintained in conditioned water alone served as controls. After hatching, larvae were fed on boiled spinach until the completion of metamorphosis. In control group, larvae that hatched on 3rd day were heavily pigmented, voracious feeders, and active swimmers; in these tadpoles, hind limb and forelimb-buds emerged on 16th and 24th day and metamorphosis was complete on 32nd day. Eggs exposed to acephate also hatched on 3rd day but larvae exhibited deformities such as, (i) tail distortions, (ii) laterally crooked trunk, (iii) decreased pigmentation, (iv) inactivity, (v) peeling of the skin, and (vi) delay in emergence of limbs and completion of metamorphosis. Cypermethrin-exposed eggs exhibited a delay (4–8 days) in hatching, there was no mortality, deformities in tail, trunk and head region, delay in the emergence of limbs, and completion of metamorphosis were evident. The demonstrated data indicate that these pesticides interfere with amphibian development when present in the aquatic system.     

锌离子慢性暴露对中华大蟾蜍蝌蚪生长发育的影响

为评价水域环境中Zn~(2+)对两栖动物的毒性影响,本研究以中华大蟾蜍(Bufo gargarizans)蝌蚪为试验材料,对G26期蝌蚪进行了0、10、50、100和500μg·L~(-1)Zn~(2+)的慢性水体暴露直至蝌蚪发育至变态高峰期(G42期)。分别于暴露15 d和30 d后取样测定蝌蚪全长、体长、体重和发育分期各指标;此外,分析了Zn~(2+)慢性水体暴露对变态率、变态高峰期(G42)蝌蚪的全长、体长、体重、后肢长和骨骼发育的影响。结果表明:暴露15 d时,各浓度Zn~(2+)对中华大蟾蜍蝌蚪的生长发育均未造成显著影响,而持续暴露30 d后,500μg·L~(-1)Zn~(2+)处理组蝌蚪的生长发育受到显著抑制;各浓度Zn~(2+)慢性暴露均导致中华大蟾蜍蝌蚪变态率的下降,其中500μg·L~(-1)Zn~(2+)处理组的变态率最低;500μg·L~(-1)Zn~(2+)慢性暴露导致G42期中华大蟾蜍蝌蚪的形态指标(全长、体长、后肢长)和骨化程度均受到显著抑制。研究表明,水环境中高浓度锌对中华大蟾蜍幼体的生长发育和变态具有潜在的危害,水域锌污染所引发的毒理效应予以重视。     

Suitability of Golf Course Ponds for Amphibian Metamorphosis When Bullfrogs Are Removed

Abstract:  Managing areas designed for human recreation so that they are compatible with natural amphibian populations can reduce the negative impacts of habitat destruction. We examined the potential for amphibians to complete larval development in golf course ponds in the presence or absence of overwintered bullfrog tadpoles ( Rana catesbeiana ), which are frequently found in permanent, human-made ponds. We reared larval American toads ( Bufo americanus ), southern leopard frogs ( R. sphenocephala ), and spotted salamanders ( Ambystoma maculatum ) with 0 or 5 overwintered bullfrog tadpoles in field enclosures located in ponds on golf courses or in experimental wetlands at a reference site. Survival to metamorphosis of American toads, southern leopard frogs, and spotted salamanders was greater in ponds on golf courses than at reference sites. We attributed this increased survival to low abundance of insect predators in golf course ponds. The presence of overwintered bullfrogs, however, reduced the survival of American toads, southern leopard frogs, and spotted salamanders reared in golf course ponds, indicating that the suitability of the aquatic habitats for these species partly depended on the biotic community present. Our results suggest that ponds in human recreational areas should be managed by maintaining intermediate hydroperiods, which will reduce the presence of bullfrog tadpoles and predators, such as fish, and which may allow native amphibian assemblages to flourish.