长江岸线宜昌江段五种不同类型水样对稀有鲫胚胎的毒性

为了检测和评估长江岸线宜昌江段五种不同类型水样的生物毒性, 研究于2019年3月在宜昌江段采集五种典型水样, 采用稀有鲫胚胎进行了暴露试验。经检测发现3#水样(工业生产污水)、4#水样(生活污水)常规污染物(氨氮、化学需氧量、高锰酸盐指数)等指数超标。胚胎毒性试验结果显示3#和4#水样的生物毒性最大, 其畸形率和死亡率等指标均显著高于其他组(P<0.05)。2#水样(长江白洋港江段水样)对稀有鲫胚胎发育无显著影响; 其中3#和4#两组水样处理的胚胎出现诸如脊椎弯曲、心率下降、死亡率上升、孵化率下降和发育迟缓等发育异常现象; 同时, 4#水样处理的胚胎还出现大量心包水肿现象。研究表明氨氮含量等常规水质指标超标会对稀有鲫胚胎产生较大毒性效应。研究可为长江岸线宜昌江段水体的综合评价及后续治理提供依据。     

稀有■鲫亚慢性毒性试验

研究了一种快速评价有毒化学品对稀有鲫慢性毒性的试验方法。选择＜3d龄的稀有峋鲫幼鱼开始暴露,试验药物为国际标准参比毒物CU( 2),Cr( 6),Zn( 2)和PCP;试验温度为25±1℃,每天更换试验液一次,试验持续5d。以现存量为测试指标,Cu( 2),Cr( 6),Zn( 2)和PCP对稀有鲫的无可观察效应浓度[NOEC]为20,2500,100,40ug／L;最低可观察效应浓度[LOEC]为40,5000,200,80ug／L,与7d亚慢性毒性试验结果相接近。试验结果表明,稀有鲫亚慢性毒性试验可从7d缩短为5d。     

聚苯乙烯微塑料暴露对稀有■鲫仔鱼生长的影响

微塑料已经成为一种新型的污染物,关于微塑料本身所造成毒性效应的研究较少。本研究以稀有■鲫Gobiocypris rarus初孵仔鱼为受试对象,研究了不同粒径(0.1μm、1μm、10μm)、不同浓度(0.055μg·L~(-1)、0.55μg·L~(-1)、5.5μg·L~(-1)、55μg·L~(-1)、550μg·L~(-1))的聚苯乙烯(PS)微塑料对稀有■鲫仔鱼存活、生长的影响以及微塑料在仔鱼体内的累积和清除情况。暴露7 d后,各暴露组仔鱼的存活率和全长与空白对照组间的差异均无统计学意义(P0.05);各暴露组仔鱼的鳃、消化道内均发现荧光微塑料,荧光强度随微塑料浓度增高而增强;经过7 d净化后,仔鱼体内PS微塑料均显著减少(P0.05)。本研究表明,环境浓度水平的PS微塑料虽可在稀有■鲫体内积累,但清除速度也较快,在个体可观察水平上未见明显的毒性效应。     

稀有鮈鲫生命早期的己烯雌酚暴露对生长发育与繁殖的影响

研究了己烯雌酚(DES)对稀有鲫(Gobiocypris rarus)生命早期暴露的影响。经10μg/L和100μg/L己烯雌酚暴露26d后,稀有鲫死亡率升高,生长发育迟缓,鱼体内卵黄蛋白原(VTG)的诱导明显。经过一段时间清水养殖后,暴露组中雌鱼比例增加,雌雄鱼生长较对照组有显著变化。雌鱼性腺发育及产卵量与对照组相比虽无显著差异,但暴露组成鱼所繁育后代与对照组相比受精率、孵化率显著下降,死亡率、畸形率明显上升。这些结果说明己烯雌酚生命早期暴露影响稀有鲫的生长发育及生殖,稀有鲫生命早期暴露实验可以用于评价水生态系统中内分泌干扰物的生态影响。     

稀有鲫热激蛋白70的分子克隆及表达特征分析（英文）

为研究稀有逗鲫(Gobiocypris rarus)HSP70用于淡水毒理学和风险评估方面的可行性,通过cDNA末端快速扩增的方法首次从稀有逗鲫肝脏中成功克隆出HSP70基因的cDNA全长,命名为GrHSP70。NCBI比对分析结果显示,GrHSP70核苷酸序列与其他鱼类的HSP70基因的核苷酸序列相似性极高,相应的氨基酸序列同源性也高(大于95%)。通过实时定量PCR获得未经污染物暴露的稀有逗鲫幼鱼体内13种组织中GrHSP70的分布以及PCP暴露后肝脏中该基因的表达图谱,结果表明GrHSP70在稀有逗鲫体内的表达呈现出组织依赖性,在性腺组织中的表达最高。此外,在不同时间﹑不同浓度的PCP暴露下稀有逗鲫肝脏中GrHSP70的表达模式呈现出显著的时间-剂量依赖效应。总之,GrHSP70可作为一种非常敏感的生物标志物,适用于中国淡水环境污染的毒理学研究及风险评估。     

东湖茶港排污口底泥对稀有鮈鲫的毒性

以东湖茶港排污口底泥复溶水为试验相,采用96h急性毒性试验和胚胎—卵黄囊吸收阶段毒性试验方法,研究了东湖茶港排污口底泥对稀有的鮈鲫毒性。结果显示,高浓度的复溶水对稀有鮈鲫胚胎、仔鱼和幼鱼具有明显的毒性效应,而胚胎—卵黄囊吸收阶段更为敏感。随着复溶水浓度的增加,稀有鮈鲫受精卵孵化率降低,仔鱼畸形率增高、成活率降低、生长减慢;对胚胎—卵黄囊吸收阶段的NOEC、LOEC和MATC分别为12.5%、25%和17.68%;对幼鱼96h LC50为69.1%。本文的研究还表明,底泥经晾晒后毒性大幅降低,暗示恢复东湖通江状态并让水位自然涨落,可能有助于缓解污染、恢复生态环境。     

稀有Ju鲫亚慢性毒性试验

研究了一种快速评价有毒化学品对稀有Ｊｕ鲫慢性毒性的试验方法。选择〈３ｄ龄的稀有Ｊｕ鲫幼鱼开始暴露,试验药物为国际标准参比毒物Ｃｕ＾＋２,Ｃｒ＾＋,Ｚｎ＾＋２和ＰＣＰ;试验温度 为２５±１℃,每天更新试验液一次,试验持续５ｄ。     

Cu~(2 )、Zn~(2 )诱导稀有鮈鲫应激蛋白质的研究

以稀有鮈鲫为材料,研究了应激蛋白质作为生物学指标的敏感性。结果表明,在无可观察效应浓度下,经5d亚慢性胁迫暴露,以Cu2+为胁迫因子,稀有鮈鲫被诱导出约54KDa的应激蛋白质;以Zn2+为胁迫因子,稀有鮈鲫被诱导出约94KDa,67KDa和40KDa的应激蛋白质。应激蛋白质有可能成为一种生物学指标运用于生态风险性早期预警。     

东湖茶港排污口底泥对稀有

以东湖茶港排污口底泥复溶水为试验相,采用96h急性毒性试验和胚胎-卵黄囊吸收阶段毒性试验方法,研究了东湖茶港排污口底泥对稀有(鱼句)鲫的毒性.结果显示,高浓度的复溶水对稀有(鱼句)鲫胚胎、仔鱼和幼鱼具有明显的毒性效应,而胚胎-卵黄囊吸收阶段更为敏感.随着复溶水浓度的增加,稀有(鱼句)鲫受精卵孵化率降低,仔鱼畸形率增高、成活率降低、生长减慢;对胚胎-卵黄囊吸收阶段的NOEC、LOEC和MATC分别为12.5%、25%和17.68%;对幼鱼96h LC50为69.1%.本文的研究还表明,底泥经晾晒后毒性大幅降低,暗示恢复东湖通江状态并让水位自然涨落,可能有助于缓解污染、恢复生态环境.     

稀有ju鲫-一种新的鱼类毒性试验材料

本文研究了稀有ju鲫(Gobiocypris rarus) 作为毒性试验材料的可行性。采用换水式试验，在硬度为200 mg/L (以CaCO[3]计)、pH7.8±0.2、温度24-25℃条件下研究了铬、铜、锌和五氯酚(PCP)对稀有ju鲫的急性毒性。重铬酸钾对2日龄稀有ju鲫的24 h和96 h和LC[50]控制范围分别263.6-334.7和115.3-178.5 mg/L (n=8)。铬、铜、锌和五氯酚对2日龄稀有ju鲫的急性毒性值(96 h LC[50])范围，从铜的52.2 μg/L到铬的52000 μg/L，毒性大小的顺序是铜>五氯酚>锌>铬。研究结果表明，稀有ju鲫有可能发展成为一种较为理想的毒性试验材料。     

Life-stage-dependent sensitivity of zebrafish (Danio rerio) to estrogen exposure

The aim of this study was to identify periods in zebrafish (Danio rerio) development when estrogen exposure has long-term consequences on reproductive capabilities at the adult stage. To this end, zebrafish were exposed to 10 ng/L ethynylestradiol (EE(2)) during three stages of gonadal differentiation: (i) the juvenile hermaphroditic stage when gonads display the morphology of an immature ovary (in our zebrafish colony this lasted from 15 to 42 days post-fertilization [dpf]), (ii) the gonad transition stage when the hermaphroditic gonad differentiates into either testes or ovary (from 43 to day 71 dpf), and (iii) the premature stage of testicular and ovarian development (from 72 to 99 dpf). The consequences of stage-specific exposure to EE(2) were assessed by determining time to first spawning, fecundity (number of eggs per female per day), fertilization success (percentage of fertilized eggs) and sex ratio of the adults. Exposure during the gonad transition period induced a delay in the onset of spawning and a significant reduction of fecundity and fertilization success, whereas exposure during the hermaphroditic stage or during the premature stage had no significant impact on the reproductive parameters of adult fish. The results from this experiment pointed to the gonad transition stage as being most susceptible to persistent effects of developmental estrogen exposure. In a second experiment, the concentration dependency of the EE(2)effects was evaluated by exposing zebrafish during the gonad transition stage (43-71 dpf) to 1.67, 3 or 10 ng EE(2)/L. Significant effects of EE(2) on adult reproduction were found with 3 and 10 ng EE(2)/L, but not with 1.67 ng/L. Histological examination of the gonads revealed that at termination of EE(2) exposure (71 dpf), all individuals in the 3 and 10 ng EE(2)/L treatment possessed ovaries. However, this feminising effect appeared to be reversible since at the adult stage (190 dpf), both fish with ovaries and with testes were found. Thus, EE(2) exposure during the gonad transition stage seems to have no persistent effect on gonad histology but on reproductive capabilities.     

Effects of octylphenol and 17beta-estradiol on the gonads of guppies (Poecilia reticulata) exposed as adults via the water or as embryos via the mother

Endocrine disrupting alkylphenolic compounds have been found in the aquatic environment, and concern has arisen over the ability of these compounds to affect the reproductive system of fish. In this study, the effects of exposure to an environmentally relevant concentration of octylphenol or 17beta-estradiol on the gonad structure of fish were examined. Viviparous guppies (Poecilia reticulata) were exposed as adults via the water or as embryos via the mother to 26 microg/l octylphenol or 0.85 microg/l 17beta-estradiol (mean measured water concentrations). Histological examinations revealed effects of the exposures on the gonads of the fish exposed as adults. Indications of blocked spermatogonial mitosis were seen in the testis structure of adult males after exposure to octylphenol or 17beta-estradiol. The post-parturition ovaries of adult females exposed to 17beta-estradiol showed effects suggesting an inhibited yolk deposition. At the tested concentrations, exposure to octylphenol or 17beta-estradiol via the mother fish did not significantly affect the weight, length, gonopodium index or sex distribution of the offspring. However, histology revealed effects on the liver structure, suggesting some effect of maternal exposure to octylphenol or 17beta-estradiol. These findings indicate that although octylphenol and 17beta-estradiol affect the gonad structure of adult male and female guppies, these substances have no significant effects on the sexual differentiation of the embryos at the tested concentrations.     

Vitellogenin in rare minnow (Gobiocypris rarus): identification and induction by waterborne diethylstilbestrol

Rare minnow (Gobiocypris rarus) is a tiny Chinese carp that has a short life cycle and is easily cultured in the laboratory. In this study, juvenile rare minnows were exposed to waterborne diethylstilbestrol (DES) at 0.05, 0.5 and 5 microg/l in laboratory aquaria. After exposure for 4, 8, 13 and 21 days, juvenile fish were collected and vitellogenin (Vtg) was measured in whole body homogenates. Native and SDS electrophoresis followed by Western blotting were performed for Vtg identification, and a non-competitive ELISA was developed. In the DES exposure groups (0.5 and 5 microg/l DES), Vtg appeared after 4 days, increased significantly after 8 days and reached a maximum on day 13. Further, a significant increase in the hepatosomatic index (HSI) was found in the 5 microg/l DES exposure group after 21 days. These results indicate that rare minnow provides a good model for assessing endocrine disruption by environmental estrogens.     

Reproductive and histopathological effects of long-term experimental exposure to bis(tributyltin)oxide (TBTO) on the three-spined stickleback, Gasterosteus aculeatus Linnaeus

Three-spined sticklebacks, Gasterosteus aculeatus Linnaeus, were exposed to 0, 0.1, 1.0, 2.5 and 10 μg bis(tributyltin)oxide (TBTO) 1⁻¹ for up to 7.5 months in a flow-through system of sea water. As early as 4 weeks after the start of the experiment, fish from the 10 μg TBTO 1⁻¹ group showed diminishing appetite, increasing lethargy and opaque eyes. The mortality in this group after 2 months of exposure was 80%. No significant differences between the control and the exposed groups were noted concerning fecundity, hatched fry and frequencies of malformed fry. The gonad somatic index (GSI) was determined before and during the spawning period. GSI in control fish increased significantly, while that in the exposed groups remained unchanged between the two sampling occasions. Structural examination of the gills from fish exposed for 2 months to 10 μg TBTO I⁻¹ revealed fusion between secondary lamellae and gradual cell degeneration. The chloride cells were characterized by degenerating mitochondria. Hepatocytes in fish exposed to 2.5 and 10 μg TBTO I⁻¹ contained an increased number of lipid-containing vacuoles and the nucleoli were disorganized and fragmented.     

Effects of the toxic dinoflagellate, Alexandrium fundyense on three species of larval fish: a food-chain approach

Sublethal behavioural effects of exposure to paralytic shellfish toxins (PST; saxitoxin and its derivatives) from the toxic dinoflagellate Alexandrium fundyense were investigated in newly settled winter flounder Pseudopleuronectes americanus , larval sheepshead minnow Cyprinodon variegatus and larval mummichog Fundulus heteroclitus through an A. fundyense –copepod–fish food chain. Consumption of as few as six to 12 toxin-containing copepods was lethal to the fishes. After consuming fewer toxin-containing copepods, all three fish species exhibited sublethal effects from vector-mediated exposure. Prey-capture ability of mummichogs was reduced in larvae that had consumed toxic copepods, Coullana canadensis . After consuming toxic C. canadensis or mixed copepods, mummichog larvae had reduced swimming performance. Swimming activity was also significantly reduced in winter flounder after consuming toxic copepods, including time spent in motion and distance travelled. Prey capture and predator avoidance were reduced in first-feeding sheepshead minnow larvae that had consumed toxic dinoflagellate cells. Adverse effects on prey capture or predator avoidance may reduce larval survival and facilitate the transmission of PST through the food web. This study provides baseline information on sublethal effects of PST exposure on fishes using a novel food-chain approach with zooplankton as vectors.     

Gyrodactylus salaris infecting allopatric Arctic charr Salvelinus alpinus fry: an experimental study of host survival

Artificially reared Arctic charr Salvelinus alpinus fry (post yolk‐sac stage) from Skibotnelva in northern Norway were exposed to infection with the monogenean ectoparasite Gyrodactylus salaris to determine if the parasite caused mortality in fry. About 95% of the fry became infected within the first week post‐exposure, and at the end of the experiment (77 days) all fry carried the parasite. Mortality rate was significantly higher in the groups of infected fry than in the uninfected control groups, and when the experiment was terminated c. 30% of the infected and 8% of the uninfected fish had died. Gyrodactylus salaris had a pronounced negative effect upon fry survival and caused an additive host mortality. Moreover, the concurrent temporal changes in parasite intensities and aggregation indicated that the parasite‐induced host mortality was density dependent.     

Observations on the biology of the perch, Perca fluviatilis, in the River Stour, Dorset

SUMMARY. Opercular bones from 323 perch from the River Stour were used for age and back-calculated growth determinations. Annuli were formed during May at the beginning of the growth period. Growth was minimal from October to April. Female perch grew faster than males, and the growth rates of both sexes were higher than those observed in most other European waters. Spawning occurred during late April and early May; male gonads began development in August and had attained their maximum weight in September, but the ovaries developed gradually from August until April. Immature perch had an annual cycle of condition with a maximum in June-July and a minimum in December-January. The condition of mature males and females was affected by the gonad cycle. The fecundity of Stour perch is expressed by the formula: log egg number = 2.40 log length (mm) - 1.34.
Approximately 25% of males were mature at age I and all were mature at age II, whereas most females did not spawn until age III. Ephemeroptera nymphs and minnow fry constituted the bulk of the diet of 0-group perch; Ephemeroptera nymphs, minnow fry and Corixidae were the most numerous items in I group perch, whereas older perch contained Corixidae and a wider range of fish prey species, although minnows were the most numerous of these.     

Molecular characterization of estrogen receptors 1, 2a, and 2b and their tissue and ontogenic expression profiles in fathead minnow (Pimephales promelas)

There are two estrogen receptor (ER) subtypes in fish, Esr1 and Esr2 (formerly ERalpha and ERbeta), and in some species the Esr2 subtype has two forms, Esr2b (formerly ERbeta1) and Esr2a (formerly ERbeta2 or ERgamma). There is little information, however, on the different characteristics and functional significance of the two receptor subtypes in fish, and this is especially relevant for understanding the disruption of ER signaling by chemicals with estrogenic activity. In this study, the full-length cDNAs for esr1 (3167 base pairs [bp]) and esr2b (2318 bp), and a partial-length (267 bp) cDNA for esr2a, were cloned and characterized in fathead minnow (fhm; Pimephales promelas), and their patterns of expression established during development and in adults. Real-time polymerase chain reaction revealed some clear distinctions in the ontogenic and tissue expression of fhm esr1, esr2b, and esr2a, suggesting different functions for each ER subtype. Fhm ERs were expressed in brain, pituitary, liver, gonad, intestine, and gill of male and female fish, esr2b and esr2a were also expressed in muscle. Fhm esr1 and esr2b were expressed predominantly in the liver, whereas fhm esr2a was expressed predominantly in intestine and was lowest expressed in liver. Responses of the different hepatic ERs in male fathead minnow exposed to 100 ng estradiol/L differed, with a significant induction (5-fold) of fhm esr1 but no effect on esr2b or esr2a expression, suggesting different mechanisms of regulation for the different ERs. The detailed characterization of ERs in fathead minnow provides the foundation for understanding the molecular basis of estrogenic disruption in fish.     

Toxicity assessment of insecticides commonly used in rice pest management to the fry of common carp, Cyprinus carpio, a food fish culturable in rice fields

Toxicity of four insecticides commonly used in rice pest management, chlorpyrifos, dimethoate, carbaryl and carbosulfan, to the fry of common carp was assessed through median lethal concentrations (LC₅₀) and in vivo inhibition of the brain acetylcholinesterase (AChE) enzyme at sublethal concentrations. The 96‐h LC₅₀ values for these four insecticides were determined to be 0.008, 26.11, 7.85 and 0.60 mg L^?1 respectively. Exposure of fish to a series of sublethal concentrations (0.5–5% LC₅₀) of each insecticide for 14 days resulted in concentration‐dependent inhibition in AChE activity in comparison with the controls. AChE activity was greatly inhibited in the fish exposed to sublethal concentrations of chlorpyrifos. Upon transfer to insecticide‐free water, AChE activities in fry exposed to 0.5 and 1% LC₅₀ concentrations of carbaryl and carbosulfan were restored to the control level within 7–21 days whereas the fish exposed to chlorpyrifos or dimethoate did not fully recover from the insecticide‐induced anticholinesterase action. Of the four insecticides tested, chlorpyrifos was the most toxic for the fry of common carp. Although dimethoate was least toxic for the fish under acute exposure, the restoration level of normal AChE activity was slower under chronic exposure in comparison with carbaryl and carbosulfan. Hence, the use of carbamates, especially carbaryl, to control insect pests of rice in rice‐cum‐carp culture systems is recommended when considering survival, restoration of the normal AChE activity and stamina of the cultured fish.     

Acute Toxicity,Respiratory Reaction,and Sensitivity of Three Cyprinid Fish Species Caused by Exposure to Four Heavy Metals

Using 3 cyprinid fish species zebra fish, rare minnow, and juvenile grass carp, we conducted assays of lethal reaction and ventilatory response to analyze sensitivity of the fish to 4 heavy metals. Our results showed that the 96 h LC₅₀ of Hg²⁺ to zebra fish, juvenile grass carp, and rare minnow were 0.14 mg L⁻¹, 0.23 mg L⁻¹, and 0.10 mg L⁻¹, respectively; of Cu²⁺0.17 mg L⁻¹, 0.09 mg L⁻¹, and 0.12 mg L⁻¹ respectively; of Cd²⁺6.5 mg L⁻¹, 18.47 mg L⁻¹, 5.36 mg L⁻¹, respectively; and of Zn²⁺44.48 mg L⁻¹, 31.37 mg L⁻¹, and 12.74 mg L⁻¹, respectively. Under a 1-h exposure, the ventilatory response to the different heavy metals varied. Ventilatory frequency (Vf) and amplitude (Va) increased in zebra fish, juvenile grass carp, and rare minnows exposed to Hg²⁺ and Cu²⁺ (P<0.05), and the Vf and Va of the 3 species rose initially and then declined when exposed to Cd²⁺. Zn²⁺ had markedly different toxic effects than the other heavy metals, whose Vf and Va gradually decreased with increasing exposure concentration (P<0.05). The rare minnow was the most highly susceptible of the 3 fish species to the heavy metals, with threshold effect concentrations (TEC) of 0.019 mg L⁻¹, 0.046 mg L⁻¹, 2.142 mg L⁻¹, and 0.633 mg L⁻¹ for Hg²⁺, Cu²⁺, Cd²⁺, and Zn²⁺, respectively. Therefore, it is feasible to use ventilatory parameters as a biomarker for evaluating the pollution toxicity of metals and to recognize early warning signs by using rare minnows as a sensor.