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.     

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)。     

鱼类双重荧光标记研究——以中华倒刺鲃幼鱼为例

为探究盐酸四环素(TCH)和茜素红S(ARS)对中华倒刺鲃(Spinibarbus sinensis Bleeker)的影响及标记效果, 丰富鱼类荧光标记模式, 研究使用TCH(100—500 mg/L)和ARS(100—300 mg/L)对中华倒刺鲃幼鱼进行双重浸染荧光标记, 实验共设置5个处理组和1个对照组, 浸染时间均为24h。结果显示, 经90d的养殖实验后, 标记鱼的耳石(包括矢耳石和星耳石)、倒刺、鳍条和鳍棘均能检测到双重荧光标记环, 且TCH产生的黄色荧光环比ARS产生的红色荧光环更接近骨质结构内部。较高浓度处理组的矢耳石(≥300 mg/L TCH和≥150 mg/L ARS)、星耳石(≥300 mg/L TCH 和 ≥200 mg/L ARS)和倒刺(400—500 mg/L TCH 和150—300 mg/L ARS)中均能检测到明显的标记环(n≥2), 但所有处理组的侧线鳞和非侧线鳞的荧光标记不明显(0≤n≤1)。经200—500 mg/L TCH和150—300 mg/L ARS处理的鳍条, 及经300—500 mg/L TCH和200—300 mg/L ARS处理的鳍棘中可以同时检测到明显的TCH和ARS的标记环(n≥2)。此外, 在整个实验中各处理组标记鱼与对照组相比, 在生长和存活率方面均无显著性差异(P>0.05)。结果表明, 使用TCH和ARS双重标记中华倒刺鲃幼鱼是可行的。双重荧光标记方法在水生生物标记回捕实验及实验性生物学研究方面具有潜在的应用前景。     

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.     

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。     

齐口裂腹鱼耳石早期生长发育与日轮特征研究

文章研究了在实验室条件下齐口裂腹鱼仔稚鱼耳石早期形态发育与生长特点、第一轮纹出现时间和轮纹沉积规律。结果表明: 在13.5-17.2℃孵化条件下,微耳石和矢耳石在出膜前形成,而星耳石于出膜后第12天出现。在仔稚鱼生长过程中,微耳石由近圆形发育成贻贝形,矢耳石经历近圆形、锲形后发育为箭矢状,星耳石形状由近圆形发育为星芒状。微耳石的前区、背区和腹区及矢耳石的背区和腹区生长呈幂函数关系,而微耳石的后区、矢耳石前区和后区生长以及两对耳石的前后区半径之和与全长均呈线性相关。在(18.50.5)℃和(15.61.1)℃条件下,50%矢耳石样本第一轮纹均在出膜后第 2 天形成(分别为出膜后18h和19h),以后每天形成一轮。微耳石和矢耳石轮纹数均与日龄呈线性相关,方程斜率均与1差异不显著(P0.05),表明两对耳石的轮纹沉积均为日周期性。这些结果为研究齐口裂腹鱼野生种群繁殖期和早期生活史特征等生态学问题提供了重要依据。     

骨唇黄河鱼耳石早期形态发育和轮纹特征研究

研究了骨唇黄河鱼仔稚鱼耳石在实验室养殖条件下的发育过程和生长特点,确证了轮纹沉积规律。结果表明,在14.0-17.8℃孵化条件下,微耳石和矢耳石在受精后96h 30min出现,星耳石在出膜后第16天出现。仔稚鱼生长过程中矢耳石形状变化较大,由出膜时的圆形发育到稳定时的箭矢状。微耳石由近圆形发育成贻贝形,其中心核位置随发育明显偏移。星耳石形状不规则,从出现时的心形发育成为星芒状。微耳石和矢耳石在前后轴方向上后区的生长快于前区（P0.05）;在背腹轴方向上,微耳石腹区的生长快于背区（P0.05）,矢耳石背区的生长快于腹区（P0.05）,两对耳石的前后区半径之和与全长均呈线性相关。微耳石和矢耳石的第1个轮纹均在出膜后第2天形成,新增的轮纹数（微耳石IL,矢耳石IS）与出膜后的天数（D）表现出显著的线性相关,方程分别为: IL=0.9911D-1.0008（R2=0.9971,n=220,P0.001）和IS=0.9925D-0.10873（R2=0.9919,n=161,P0.001）,方程的斜率与1均无显著差异（P0.05）,表明两对耳石轮纹沉积均呈日周期性,生长轮为日轮。研究结果丰富了骨唇黄河鱼的发育生物学资料,可为研究其自然种群早期生活史提供参考。       

鲤稚鱼耳石锶标记效果

对鲤稚鱼开展了不同浸泡浓度和不同标记时间梯度的SrCl₂·6H₂O暴露标记试验,以确认其耳石Sr标记的可行性及基本条件.首先,基于4个浓度(0、4、8、12 mg·L^-1)水平的SrCl₂·6H₂O溶液,浸泡标记2 d来筛选基本浸泡标记浓度.然后,在SrCl₂·6H₂O为8 mg·L^-1浓度下,基于5个浸泡时间(1、2、3、4、5 d)来筛选基本浸泡标记时间.电子探针分布分析结果显示:对照组(0 mg·L^-1)耳石Sr/Ca比值低且稳定,标记组均出现了高Sr/Ca比值区.对照组耳石剖面为均一的低Sr蓝色图谱,而标记组耳石上均有高Sr红色标记环带,且标记成功率为100%.试验期间,标记组和对照组的死亡率、平均全长和平均体质量无显著差异,表明Sr标记对供试鱼无不良影响.由于耳石上清晰、完整的高Sr红色标记环带出现在标记浓度为8 mg·L^-1及以上,标记时间为2 d及以上,故建议分别选择8 mg·L^-1和2 d为基本浸泡标记浓度和基本浸泡标记时间.本研究证实耳石Sr标记技术对鲤稚鱼具有很好的可行性.     

Otolith development and daily increment formation in larvae of the Kabyabya,a Malawian cyprinid, <Emphasis Type="Italic">Opsaridium tweddleorum</Emphasis>

Features of the sagitta, asteriscus, and lapillus of laboratory-hatched larvae of the Kabyabya, Opsaridium tweddleorum, were investigated until day 33 after hatching. The sagitta was round until day 17, subsequently becoming arrowhead-shaped with the development of anterior and posterior rostra. Increments in the sagitta increased at the rate of one per day before rostrum formation. However, they were occasionally overestimated in rostrum sections as a result of subdaily increment occurrence. Furthermore, both anterior and posterior rostra were fragile, being thin and platelike, and were often damaged during the extraction and grinding processes. The lapillus was also round until day 17, thereafter becoming fan-shaped. Increments in the lapillus were distinctive from the core to the margin, usually increasing at the rate of one per day after hatching. The asteriscus appeared in fish larger than 9.15 mm in standard length (from day 17), being oval with a somewhat ambiguous core, causing difficulty in discerning the first increment. These features indicated that the lapillus was the most appropriate otolith part for daily increment analysis in this species, although the sagitta was useful before rostrum formation.     

Otolith morphology, microstructure and ageing in the hedgehog seahorse, Hippocampus spinosissimus (Weber, 1913)

The life history characteristics of many species of seahorse, including the hedgehog seahorse Hippocampus spinosissimus, make them sensitive to exploitation. Consequently, proper management tools must be employed; these are often based on reliable ageing. The aim of the present study was to evaluate the possibility of using otoliths for ageing hedgehog seahorses from Vietnam. Asteriscus, lapillus and sagitta all showed moderate to strong correlations between fish standard length (SL), otolith length and number of increments in seahorses between 83 and 188 mm SL. There were no annual or seasonal marks in any of the three otolith pairs observed using dissecting or light microscopy, and only asteriscus revealed microincrements under light microscopy. The number of increments ranged between 71 and 137. A full trajectory of lapillar microincrements was only visible when using scanning electron microscopy, and then only in two of the examined individuals (88 and 115 increments). A validation experiment showed that the number of increments in the asterisci did not correspond to the age of seahorses bred and reared for 717 and 868 days. Furthermore, a second validation study using Alizarine Complexone (ALZ) otolith marking did not reveal any increments peripheral to the ALZ mark 30 days after marking. The conclusion of the study was that otoliths do not at present provide a reliable tool for estimating age in adult hedgehog seahorses, and therefore other tools have to be employed to improve their management.     

Otolith microstructure of Japanese sea bass larvae and juveniles: interpretation and utility for ageing

This paper interprets and discusses the usefulness of otolith microstructure for ageing Japanese sea bass ( Lateolabrax japonicus ) larvae and juveniles. Samples were collected from the Tango Sea along the Japan Sea coast, January–March 2007. Known-age (0-day and 10-day-old) larvae were obtained from the Ibaragi Prefectural Hatchery, Japan. Sagittal and lapillar otolith were processed and read using an otolith reading system. Clearly discernible hatch- and first-feeding marks were evident on sagitta, and development of accessory premordia (AP) appeared to be associated with larva-juvenile transition; however, no other marks indicating metamorphosis or settlement were evident. In lapillus, no discernible check mark was found. Known-age larvae showed that deposition of the first daily increment (DI) corresponded to first-feeding, which occurred at day-4 post-hatch. However, mean increment counts were significantly lower in lapillus than in sagitta, caused by poorly expressed increments around the centrum as well as relatively unclear centrum of the lapillus. The authors suggest that the use of lapillus can cause significant underestimation of age. Therefore, the sagitta is recommended for age and growth estimations of larvae and juveniles, although the presence of numerous subdaily increments warrants careful preparation and interpretation of the microstructure. A test for asymmetry showed the right and left otoliths to be quite symmetrical and their DI counts not significantly different, suggesting that either otolith can be used for studying age and growth of Japanese sea bass larvae and juveniles.     

Otolith features and utility of lapillus for daily increment analysis in <Emphasis Type="Italic">Opsaridium microcephalum</Emphasis> (Cyprinidae) juveniles collected from Lake Malawi

 Features of otoliths in Opsaridium microcephalum juveniles were observed, and the increment formation pattern was validated using Alizarin Complexone (ALC). The sagitta was arrowhead shaped with an obvious core and rostra, the latter being fragile and easily destroyed by extracting and grinding processes. Increments around the core were readable but not in the rostra. The asteriscus was oval shaped with an ambiguous core leading to difficulty in discerning the first increment. The lapillus was round and fan shaped with an obvious core. Increments in the lapillus were clearly deposited from the core to the margin. These features made the lapillus the most appropriate for reading otolith increments. Increments counts in the lapillus formed after ALC treatment agreed with days elapsed, showing the daily formation of increments. Consequently, the lapillus is only appropriate for daily increment analysis in Opsaridium microcephalum. Received: December 4, 2001 / Revised: August 14, 2002 / Accepted: September 11, 2002 Acknowledgments The authors express great thanks to J.S. Likongwe and E. Kaunda, Department of Aquaculture and Fisheries Science, Bunda College of Agriculture, University of Malawi, for their advisory comments. The authors are also grateful to the Japan International Cooperation Agency (JICA) for giving us an opportunity to work on this subject. Correspondence to:Shinsuke Morioka     

Preliminary examination on lapillus utility for otolith increment analysis in Malawian cyprinid <Emphasis Type="Italic">Engraulicypris sardella</Emphasis>

Features and increments of otoliths in Engraulicypris sardella were observed. The sagitta was arrowhead shaped with an obvious core and rostra, the latter being fragile and easily destroyed by extracting and grinding processes. Increments around the core were readable but not in rostra. The lapillus was round fan-shaped with an obvious core. Increments in the lapillus were clearly deposited from the core to the margin. The asteriscus had an ambiguous core that led to difficulty in discerning the first increment and had fewer increments than the lapilli. Thus, the lapillus is the most appropriate for reading otolith increments.     

Age and growth information available from the otoliths of the Hawaiian snapper,Pristipomoides filamentosus

The otoliths of tropical fish may provide important life history information incorporated within their structural and chemical constituents. All three otoliths (sagitta, lapillus, asteriscus) of the tropical fish Pristipomoides filamentosus were examined internally by Scanning Electron Microscope methods to observe micro-increments and externally to determine three dimensional structure. It was discovered that the sagitta contained four cores and that the plane chosen to be sectioned for micro-increment enumeration could result in errors if more than one core were transversed. The medial cross sectional plane was consequently resolved to effer the most accurate micro-increment counts. Obserations of lapilli also revealed micro-increments and subsequent counts were closely correlated to those detected in the sagittae. The visualization of increments made it feasible to assess age and evolve a growth model. In addition, sagitta weight was found to be related to growth rate and may provide a quick estimate of relative growth. Chemical analyses of otoliths for stable isotopes and Sr/Ca ratios all suggested that an individual fish inhabited warmer waters as it became older. A combination of otolith structural and chemical information can provide age and growth data which is essential to the calculation of accurate population parameters.     

大麻哈鱼胚胎耳石微结构及其群体环境标记

采用人工调控环境方法对黑龙江、绥芬河大麻哈鱼发眼期胚胎群体耳石日轮进行周期性持续标记。实验分４组: 1组为对照组, 2－3组为变温标记组, 4组为“暴气”-变温标记组, 实验用发眼卵1.2万粒。待胚胎发育至耳石日轮结构形成后实施标记。实验胚胎耳石随着标记期间环境周期性变化及其持续的时间, 形成相应变化节律的日轮标记区。获得各实验组设定环境的日轮标记图谱。人工环境标记的耳石日轮图谱, 暗带色度加深, 明带亮度增大, 并可形成生长轮距不同的标记轮, 与对照组耳石微结构有明显区别, 标记率达到100%。初步建立鱼类耳石标记及其识别技术, 适用于大麻哈鱼等鲑鳟鱼类群体标记。作为安全有效、成本低廉的群体标记技术方法, 鱼类耳石日轮标记在鱼类资源评估和增殖放流效果评价中将会得到广泛应用。