鲫幼鱼耳石荧光标记效果

为确认茜素络合物对鲫（Carassius auratus）耳石进行有效标记的可行性,以便为鲫甚至其他鲤科鱼类标志放流技术的开发及效果评估提供一定的借鉴,本研究以孵出后90 d的鲫幼鱼为研究对象,设置单一浓度（100 mg/L）的茜素络合物浸泡标记5 d,分析茜素络合物在耳石上的沉积情况以及不同耳石在不同后续饲养天数的动态变化。结果表明,矢耳石、微耳石和星耳石上在可见光、绿色和蓝色激发光下都检测到了良好的茜素络合物标记环,标记率和存活率均为100%。但不同耳石的茜素络合物标记效果不同,荧光下,星耳石的标记效果最显著,微耳石次之;可见光下,微耳石的标记效果最好,星耳石次之。随着后续饲养天数的延长,可见光下标记逐渐减弱,至20 d时基本消失,而在绿色和蓝色激光下标记环荧光强度无减弱迹象,能长久保持,且在蓝色激发光下标记环更易被观测到。上述结果结合鲫生长、存活和行为正常等情况综合显示,在100 mg/L茜素络合物溶液中浸泡标记鲫幼鱼5 d,其耳石可以获得满意的标记效果。     

滇池金线鲃(Sinocyclocheilus grahami)耳石的茜素红及茜素络合物标志

该文使用不同浓度梯度(50~200 mg/L)和浸泡时间(4~24 h)的茜素络合物(ALC)及茜素红(ARS)水溶液对6、20及90日龄滇池金线鲃(Sinocyclocheilus grahami)仔稚鱼进行标志。两种标志物均能在其耳石上形成深红色标记环带,510~560 nm绿色激发光下,标志带呈猩红色荧光反应。50 mg/L ALC或ARS溶液浸泡6日龄仔鱼8 h、50 mg/L ALC溶液或100 mg/L ARS溶液浸泡20日龄仔鱼24 h及100 mg/L ALC或150 mg/L ARS溶液浸泡90日龄稚鱼24 h,可见光下即可见清晰深红色标记带。50 mg/L ARS或ALC溶液浸泡6日龄仔鱼4 h或浸泡20、90日龄仔鱼24 h,绿色激光下即绿色可见猩红色标记带。且以上浸泡条件均可保证100%标记率和存活率。     

用荧光物质浸泡标记胭脂鱼仔、稚鱼耳石

用茜素络合物和盐酸四环素溶液分别对胭脂鱼（Myxocyprinus asiaticus）仔、稚鱼浸泡标记,结果表明,100～200mg／L的茜素络合物溶液浸泡24h对胭脂鱼仔、稚鱼耳石有很好的标记效果。相同浸泡浓度下,随着日龄的增长,耳石上的荧光反应强度降低。三对耳石中,微耳石和矢耳石对茜素络合物较敏感,星耳石敏感性则较低。盐酸四环素溶液对仔、稚鱼耳石的标记效果很差,浸泡液浓度为100mg／L和120mg/L时,仅能在微耳石上检测到较弱的荧光标记,而150mg／L及以上浓度的浸泡液对稚鱼有较高的致死作用。因此,茜素络合物是对胭脂鱼早期鱼苗进行化学标记比较合适的荧光物质,而盐酸四环素不适于标记该鱼的耳石。在对标记鱼进行荧光检测时,矢耳石和微耳石是适合的材料。     

用荧光物质浸泡标记重口裂腹鱼仔鱼耳石

用荧光物质对重口裂腹鱼仔鱼浸泡标记表明,200～300 mg/L的茜素络合物溶液浸泡12 h对重口裂腹鱼仔鱼耳石有很好的标记效果,荧光和可见光下均能检测到明显的标记环;100～150 mg/L的标记能检测到荧光标记环,但可见光下标记环较微弱.相同浸泡时间,随着浸泡浓度的增加,耳石上标记环强度增加;相同浸泡浓度,随着浸泡时间的增加,耳石上标记环强度增加.3对耳石中,微耳石和矢耳石对茜素络合物较敏感,星耳石敏感性较低.盐酸四环素的标记效果很差,浸泡浓度为50～160 mg/L时,3对耳石上均不能检测到标记环,同时110～160 mg/L的盐酸四环素浸泡液对仔鱼有较高的致昏或致死作用.因此,茜素络合物是对重口裂腹鱼鱼苗进行化学标记比较合适的荧光物质,而盐酸四环素不适合于标记该鱼的耳石.     

唐鱼仔鱼耳石的形态发育及日轮

观察了实验室人工繁殖饲养的唐鱼(Tanichthys albonubes)仔鱼耳石形态发育,研究了其生长轮的沉积规律。唐鱼仔鱼耳石长径与鱼体全长(TL)均呈线性相关,其关系式为:微耳石Dl=0.019 6TL-0.031 0(r=0.961 6,P<0.001,n=218),矢耳石Ds=0.027 6TL-0.043 7(r=0.924 0,P<0.001,n=219),星耳石Da=0.016 6TL-0.004 1(r=0.369 6,P<0.001,n=44)。仔鱼耳石上第一个轮纹在孵出后第2 d形成,生长轮数目与仔鱼日龄(D)呈线性相关,其斜率与1无显著差异,因此生长轮为日轮,其关系式为:微耳石LI=1.006D-1.700 1(r=0.994 2,P<0.001,n=205),矢耳石SI=0.953 8D-0.911 6(r=0.993 5,P<0.001,n=161)。生长过程中矢耳石形状变化较大,星耳石出现时间较晚,而微耳石形状稳定,日轮可读性较好,故更适合作为日轮研究的材料。     

鲈鲤早期鱼苗的耳石标记研究

为研究鲈鲤早期鱼苗耳石标记的可行性,先采用高温水(20.8±0.3)℃和低温水(10.8±1.2)℃交替饲养对20日龄鲈鲤进行热标记,然后将经热标记的35和45日龄鲈鲤浸泡在50—200 mg/L的茜素络合物(AC)或茜素红S(ARS)溶液中进行荧光标记。热标记的耳石生长轮清晰可见,明显区别于其他轮纹,微耳石热标记轮轮纹宽度(IW)和高温期持续时间(T)的线性回归方程为IW=0.16462+0.24762T。经荧光物质浸泡后鲈鲤耳石在绿光下均能检测到橘红色标记; AC标记质量受溶液浓度影响显著(P<0.05),受全长的影响不显著(P>0.05),微耳石、矢耳石和星耳石间的标记质量差异显著(P<0.05); ARS标记质量受全长和溶液浓度影响不显著(P>0.05),微耳石、矢耳石和星耳石间的标记质量差异显著(P<0.05)。研究结果表明,耳石热标记、荧光标记及两者结合使用均可用于大规模标记鲈鲤早期鱼苗。     

荧光物质浸泡标记稀有鲫和彭泽鲫仔、稚鱼

研究了茜素络合物和盐酸四环素对稀有Ｊｕ鲫和彭泽仔,稚鱼进行浸泡标记的方法和效果。茜素络合物在浸泡浓度５０－１００ｍｇ／Ｌ,浸泡时间持续６－２４ｈ的范围内,对两种鱼都有很好的标记效果;盐酸四环素在浸泡浓度１５０－２５ｍｇ／Ｌ,浸泡时间１２－２４ｈｘ的范围内,对彭泽鲫有较好的标记效果,但对稀有Ｊｕ鲫有极高的死率。在荧光显微镜下观察,茜素络合物呈橘红色,盐酸四环素标记区呈金黄色荧光反应。     

草鱼仔鱼耳石的自然标记和生长轮的清晰度

将野生和人工繁殖的草鱼仔鱼的耳石取出并置于显微镜下观察微结构特征。结果表明 :草鱼耳石一般有一个圆形或卵圆形的原基和中心核 ,但有 0 4 1%～ 4 6 7%的样本具有双原基或双中心核。343尾野生仔鱼中 ,6 71%的个体在矢耳石和微耳石上具有营养转换标记 ,而 187尾人工繁殖的仔鱼中 ,在矢耳石和微耳石上出现营养转换标记的比例分别是 6 4 17%和 5 0 80 % ;在营养转换标记处 ,矢耳石和微耳石的直径分别为 5 4 12±9 4 9μm和 4 0 4 8± 7 0 2 μm (n =5 0 ) ;133尾野生仔鱼在转入实验室饲养的过程中 ,86 4 7%的个体在耳石上形成了转移标记 ;野生仔鱼生长轮纹清晰的矢耳石 (n =5 2 1)和微耳石 (n =5 2 1)样本的比例分别低于 10 %和2 5 % ,但在人工饲养仔鱼中 ,95 0 0 %个体的矢耳石 (n =186 )和 88 0 0 %个体的微耳石 (n =184 )具有清晰的生长轮纹 ;野生仔鱼经人工饲养后 ,其耳石上在饲养期间沉积的生长轮的清晰度亦明显比在野外生存期间沉积的高 ;对比实验显示饥饿对仔鱼耳石生长轮的清晰度没有明显的影响.     

切鳍标记对唐鱼游泳能力的影响

本研究通过测定完全切除背鳍、臀鳍、尾鳍或双侧胸鳍后唐鱼（Tanichthys albonubes）临界游泳速度来评价切鳍标记对游泳能力的影响。研究结果表明,在速度增量（ΔU）为36 mm/s,持续时间（Δt）分别为5 min、10 min、15 min、20 min、25 min、30 min的条件下,唐鱼临界游泳速度会随着持续时间的延长而降低。鳍组织完整的唐鱼〔体长（24 ± 1）mm〕,在不同持续时间（Δt）条件下,其绝对临界游泳速度（Uacrit）为（251.98 ± 11.04）mm/s ~（333.78±12.44）mm/s;同等条件下,切除唐鱼的背鳍或臀鳍均不会对唐鱼的绝对临界游泳速度造成显著影响（P＞0.05）,但切除唐鱼的尾鳍或胸鳍后其临界游泳速度与对照组相比极显著降低（P＜0.01）,其中切除尾鳍后绝对临界游泳速度平均下降47.20%,切除胸鳍后平均下降22.98%,两者间存在极显著的差异（P＜0.01）(图2)。研究表明背鳍切除可作为野外唐鱼短期标记-重捕的手段之一。     

鳡鱼仔稚鱼耳石的标记和其日轮的确证

用茜素络合物对鳡鱼仔鱼进行了浸泡标记,耳石上能检测到橘红色荧光标记环。100mg/L茜素络合物溶液的标记效果较差;120mg/L和150mg/L溶液浸泡后微耳石的标记率很高,矢耳石和星耳石的标记率低。标记后耳石上的生长轮数与饲养天数间呈一一对应关系,相关方程为:N=0.2663 0.9276D(n=68,r~2=0.9664)。方程的斜率0.9276与1无显著差异,证明生长轮确系日轮。鱼体长与微耳石、矢耳石及星耳石的直径间呈显著的线性关系,相关方程为:BL=66.8723LD 2.7064(n=73,r~2=0.8867),BL=22.7839SD 6.6066(n=49,r~2=0.8525),BL=47.6079AD 3.5660(n=71,r~2=0.9012)。     

唐鱼精巢的组织学观察

应用光学显微镜对唐鱼Tanichthys albonubes精巢的组织结构进行了观察.结果表明,唐鱼的精巢属于小叶型结构.性成熟唐鱼的精巢呈乳白色,长条状,左右各一,合并成“Y”型.小叶间质把精巢分成许多精小叶,每个精小叶由数个精小囊组成,精子就在精小囊中形成.同一精小叶内的精小囊不一定同步发育,但同一精小囊中的生精细胞发育是同步的.唐鱼的精子发生和形成过程经历了初级精原细胞、次级精原细胞、初级精母细胞、次级精母细胞、精子细胞和成熟精子6个阶段.精巢内同时存在初级精原细胞和次级精原细胞两种类型的精原细胞.     

唐鱼肝脏显微和超微结构观察

应用光镜和透射电镜对繁殖期间唐鱼Tanichthys albonubes肝脏组织的显微和超微结构进行了观察。结果显示,唐鱼肝细胞具单核,中央核仁显著;细胞质内分布着粗面内质网、线粒体、糖原颗粒和脂滴等细胞器和内含物。胆小管由2～3个相邻肝细胞质膜凹陷围成,而肝血窦则由内皮细胞的胞质、成纤维细胞等参与构成。肝细胞与周边细胞通过3种不同方式进行联系:肝细胞之间的紧密连接;与血窦的间接连接;与胆小管的邻接。这些联系方式显示了肝脏具有内分泌腺和外分泌腺功能的特点。研究还发现雌性唐鱼肝脏具有"暗"细胞和"淡"细胞两种类型。本文还讨论了唐鱼肝脏与其他硬骨鱼类肝脏一般组织结构和超微结构的异同点。     

唐鱼卵黄蛋白原的ELISA检测方法的建立

目的探索建立唐鱼卵黄蛋白原的ELISA检测方法。方法以卵黄脂磷蛋白（Lv）抗血清为抗体,以纯化的Lv作为抗原建立间接酶联免疫吸附反应（ELISA）方法检测雄性唐鱼（Tanichthys albonubes）整体匀浆液中的卵黄蛋白原（Vtg）。结果利用ELISA方法测定了经17-β雌二醇（E2）和不同浓度DDTs暴露21 d诱导的雄鱼整体匀浆液中的Vtg含量,可以直接在1块或在不同的酶标板上准确地进行比较。经0.055 mg/mL E2诱导的雄鱼整体匀浆液中Vtg含量为4148.33μg/g;当暴露DDTs浓度为0.0275、0.0137和0.0067 mg/mL时,雄鱼整体匀浆液中Vtg含量分别为1109.43、911.16和1322.79μg/g,与丙酮溶剂对照组462.79μg/g比较差异存在显著性（P〈0.05）。结论建立了唐鱼卵黄蛋白原的ELISA检测方法 ,本方法的检测灵敏度为8.1 ng/mL,批内误差为8.59%,批间误差为6.28%,工作范围为3.26～209.25 ng/mL。在该范围内,标准曲线具有良好的线性和重复性。     

Threatened fishes of the world: Tanichthys albonubes Lin 1932 (Cyprinidae)

The white cloud mountain minnow Tanichthys albonubes Lin is an endemic species to southern China and the genus has two species, Tanichthys albonubes Lin and Tanichthys micagemmae Freyhof et Herder. The distribution range of T. albonubes Lin and T. micagemmae is very narrow and only found in the mountain brooks of Baiyunshan Mountain (White Cloud Mountain), Huaxian Country and the vicinity of Guangzhou in Guangdong Province and Halong, Quang Ninh Province, Vietnam respectively. The wild populations of this fish had already been on the verge of extinction when Shu-Yan Lin first discovered it in 1932 at the Baiyunshan Mountain. It was believed to be extinct in the wild because there were no reports of this fish in the wild since 1980. In September 2003, a small and isolated population of the fish was discovered in a mountain puddle in the north vicinity of Guangzhou. Additional studies are needed to determine the survival and propagation of the released fish. The protection of their natural habitat should be implemented.     

铜、锌、镉对唐鱼胚胎及初孵仔鱼的急性毒性及安全浓度评价

采用静水法生物测试研究铜（Cu2＋）、锌（Zn2＋）和镉（Cd2＋）对唐鱼（Tanichthys albonubes）胚胎和初孵仔鱼的急性毒性及安全浓度评价。结果显示,Cu2＋对唐鱼胚胎12、24h LC50分别为2.4092mg/L和0.4039mg/L;Zn2＋和Cd2＋对唐鱼胚胎24h LC50分别为372.9mg/L和50.0mg/L。Cu2＋对唐鱼初孵仔鱼12、24和48h LC50都是0.3228mg/L,其安全浓度为0.0986mg/L;Zn2＋对唐鱼初孵仔鱼24、48h LC50分别为72.44mg/L和25.17mg/L,其安全浓度为0.9116mg/L;Cd2＋对唐鱼初孵仔鱼24、48h LC50分别为36.5mg/L和20.59mg/L,其安全浓度为1.9654mg/L。结果表明,重金属元素对唐鱼胚胎和初孵仔鱼毒性依次为Cu2＋〉Cd2＋〉Zn2＋。     

Performance of marking techniques in the field and laboratory for Diabrotica speciosa (Germar) (Coleoptera: Chrysomelidae)

A reliable marking technique was needed for a mark–release–recapture experiment with adults of Diabrotica speciosa (Germar). Four marking techniques, acrylic paint (spattered or brushed on the surface of the insect); and fluorescent pigments (dusted on surfaces or mixed with diet to produce an ingested marker), were tested. Fluorescent pigment durability for the dusting and ingested techniques was evaluated for laboratory conditions and under simulated field conditions. The impact of the techniques on beetle survival was also assessed. Both acrylic paint techniques caused mobility problems in the beetles, and neither technique lasted for more than 48 h. Both fluorescent pigment techniques were more reliable, but the dusting technique showed a significantly higher mortality than the control, and duration variations between laboratory and field conditions. Use of fluorescent pigments added to the diet was the most reliable technique. This technique allowed the manipulation of the marking period, and provided reliable timing of marker persistence in the field.     

Validation of annual growth bands in the otolith of tropical parrotfishes (Scarus schlegeli Bleeker)

The otoliths of the scarid Scarus schlegeli (Bleeker) from the Lizard Island on the northern Great Barrier Reef showed a series of clear periodic bands. The rate at which bands are deposited on the otoliths was distinguished by mark-recapture experiments using tetracycline and marginal increment analysis. Mark-recapture experiments were carried out both in the field and in a large aquarium system. One otolith band was deposited during the liberty periods of July 1990 to April and September 1991 while two similar bands were formed from November 1989 to February 1991. The marginal increment analysis indicated that annual bands were formed around the austral summer. Scales also displayed regular period check marks. These provided relatively reliable aging estimates for the younger fishes up to 5 years.     

The use of otolith shape and chemistry to determine stock structure of Mediterranean horse mackerel Trachurus mediterraneus (Steindachner)

Otolith shape and chemistry of Mediterranean horse mackerel Trachurus mediterraneus were simultaneously used to assess the feasibility of using these natural tags to discriminate populations throughout the Black, Marmara, Aegean and eastern Mediterranean Seas. Otolith shape and chemistry analyses showed a similar pattern of differentiation between T. mediterraneus stocks, revealing a clear discreteness of the middle Black Sea (Sinop) and Aegean Sea (Izmir) samples. Otolith upper side length and width, and Na, K, Mg and Ba, were the morphological traits and trace elements, respectively, differing most among groups. Overall assignment of individuals into their original sample was high for both otolith shape and chemistry. Highest reclassification rate was observed for the south-middle Black Sea and Aegean Sea samples for both analyses. Hierarchical cluster analyses also supported high differentiation of the south-middle Black Sea and Aegean Sea samples for both analyses. Mantel's test revealed that the Euclidean distance both for otolith shape ( r =−0·0917, P > 0·05) and chemistry ( r =−0·1248, P > 0·05) between these populations were not significantly associated with their geographical distances.     

Intrastream variation in growth rates and time of first otolith increment formation for young of the year Etheostoma simoterum (Cope) (Perciformes: Percidae)

Growth rates and otolith-estimated hatching dates of young of the year Etheostoma simoterum were studied at four localities along an elevational gradient in the Little River, Tennessee, U.S.A. during the summer of 1983. Changes in otolith increment width and mean daily air temperature corresponded well with each other, suggesting that increments were laid down daily. Growth rate increased nearly threefold from the highest elevation site to the lowest. Median otolith-estimated hatching dates differed by about 2 weeks between sites, and the pattern was not directly related to elevation. These data indicate that within-stream, life-history variation of a broadly distributed species should be taken into consideration when studying factors regulating stream fish populations along elevational gradients.     

The effect of social environment on estrogen excretion,scent marking,and sociosexual behavior in tamarins (Saguinus oedipus)

The influence of various social environments on estrogen excretion, scent marking, and the expression of sociosexual behavior was examined in cotton-top tamarins (Sanguinus o. oedipus). Behavioral observations and urine collections were conducted on five females while first housed in their natal family group or in the presence of another cycling adult female and then while housed with an unrelated adult male in a separate cage. Behavioral observations only were conducted on males in natal family groups and while housed with an unrelated adult female. Levels of urinary estrone and estradiol for females were measured by specific radioimmunoassays. Females housed in family groups or in the presence of an adult cycling female showed low and noncyclic patterns of estrogen excretion. Removal from these environments resulted in a rapid increase in urinary estrone and estradiol and three of five females demonstrated cyclic patterns of estrone excretion. Rates of anogenital marking in females were elevated after the social change, and sexual interactions, virtually absent in the original environments, were observed in all five females. In males, similar manipulations of the social environment affected the expression of sexual behavior, but not scent marking. The social environment, therefore has a profound impact on fertility and sociosexual behavior in cotton-top tamarin groups, with implications for callitrichid social structure and behavior.