茜素络合物对鲤仔鱼耳石标记特征研究

利用100 mg/L的茜素络合物（alizarin complexone,ALC）对鲤仔鱼进行48 h的水环境浸泡标记,以探讨该ALC标记方法的特征,及其对矢耳石、星耳石和微耳石的标记效果以及鱼体ALC浸泡、续养恢复与耳石ALC标记区域形成和消失的时滞进行研究。结果显示,3种耳石在可见光和荧光下均能检测到明显的标记环。其中星耳石的标记效果最佳,微耳石次之。耳石上荧光信号出现和消失的时间与鱼体ALC浸泡开始和结束的时间均存在1 d的时滞。此外,浸泡标记过的鲤仔鱼在进行了长达50 d的续养恢复后,其耳石上的ALC标记环仍清晰可见。研究表明,ALC标记法所形成的标记环在耳石上可长期存在,使用ALC对鲤仔鱼进行生态标记具有很强的可行性。     

茜素络合物对鳙耳石的标志效果研究

利用不同浓度梯度(50~150mg/L)的茜素络合物(ALC)水溶液对20日龄的鳙幼鱼浸泡8h进行标志。结果显示,ALC溶液浓度不高于100mg/L时,鳙幼鱼存活率为100%,150mg/L浓度下幼鱼全部死亡。茜素络合物在其耳石上能形成标记环带,随着ALC溶液浓度的增加,在可见光下标记色越来越明显,50mg/L ALC溶液中幼鱼耳石略见淡红色标记,75mg/L浓度组有清晰的淡红色标记环,100mg/L ALC溶液中幼鱼标记环带明显,鳙幼鱼最适的ALC标记浓度为100mg/L。     

茜素络合物在青鱼幼鱼耳石上的标记效果评价

放流标记技术研究是支撑增殖放流效果评估的重要内容。该研究用质量浓度为100 mg·L-1的茜素络合物(ALC)对约2月龄的青鱼(Mylopharyngodon piceus)幼鱼进行48 h浸泡,随后移入正常水体中恢复续养20 d。对其3种耳石在可见光和不同的激发光下进行比较分析,评价了茜素络合物在不同耳石上的沉积情况与标记效果。结果表明,通过在可见光和不同的激发光下进行观察,其3种耳石上均可见明显的标记区,标记率达100%。在对标记鱼恢复续养20 d后发现,耳石上的荧光标记区强度仍很高,表现出了较好的标记效果。此外,相关标记的最佳检测光源为蓝色激发光,绿色激发光次之。微耳石是用于青鱼幼鱼耳石荧光标记观察的最适类型,同时也是用于时滞检测的最适类型,其次是星耳石。此外,将鱼体浸泡入茜素络合物溶液中至其在耳石上形成标记沉积,以及将鱼体移出浸泡溶液至其耳石上标记区域消失,均存在1 d时滞。     

茜素络合物浸泡标记秦岭细鳞鲑发眼卵及仔鱼耳石

采用茜素络合物(ALC)在秦岭细鳞鲑(Brachymystax lenok tsinlingensis)发眼卵和仔鱼阶段进行浸泡标记试验,为确定合适浸泡浓度和持续浸泡时间,试验设置了6个的浓度组和4个时间梯度组.结果显示:在荧光显微镜下观察被浸泡的仔鱼和经浸泡处理的发眼卵而出膜后的仔鱼,其耳石都出现橘红色荧光标记.浸泡...     

草鱼仔鱼耳石茜素络合物标记特征研究

草鱼(Ctenopharyngodon idellus)是我国主要增殖放流经济鱼种之一,探索有效的标志方法对其增殖放流的技术保障和效果评价无疑至关重要。本研究对草鱼仔鱼耳石进行了1 d茜素络合物(ALC)荧光标记,以确认该方法的可行性及适宜条件。结果显示,微耳石、矢耳石和星耳石均能被标记。标记环在可见光及荧光下均能被观察到。其中,后者使用蓝色激发光(WBS)观察效果最佳。总体上,微耳石的标记效果最好,矢耳石次之。ALC浸泡对鱼体的生存及生长无显著影响,耳石上荧光信号出现和消失与鱼体开始浸泡和结束均存在约12 h的时滞。浸泡标记过的实验鱼在经过长达60 d的恢复续养后,标记环仍清晰可见,表明ALC标记效果明显且可长期存在。该方法对草鱼仔鱼的增殖放流标记具有较强的可行性。     

茜素络合物对半滑舌鳎(Cynoglossus semilaevis)苗种耳石的染色标记效果

采用茜素络合物(Alizarin Complexone,ALC)分别浸泡标记70 d和170 d半滑舌鳎(Cynoglossus semilaevis)苗种耳石,设置了5个浓度组和3个时间梯度组.结果显示,ALC对半滑舌鳎70-170 d苗种的耳石染色效果明显.综合考虑存活率和标记效果,ALC浸泡标记半滑舌鳎苗种耳石的适宜浓度为100 mg/L (36 h浸泡处理)和150 mg/L(24 h浸泡处理).相同浸泡时间下,随着ALC浓度的增加,耳石上标记信号强度增加,但实验鱼死亡率升高;相同浸泡浓度下,随着时间的增加,耳石上标记信号强度增加,实验鱼死亡率也会升高.对标记的170d苗种进行养殖实验,每30 d取样矢耳石进行观察,发现4个月后实验鱼生长良好,且耳石荧光信号仍可清晰检测到,表明ALC在半滑舌鳎矢耳石上形成的荧光标记可长期存续,标记效果好.研究结果可为半滑舌鳎大规模标志放流及早期生活史研究提供群体身份识别技术支持.     

茜素络合物对半滑舌鳎(Cynoglossus semilaevis)苗种耳石的染色标记效果

采用茜素络合物(Alizarin Complexone,ALC)分别浸泡标记70 d和170 d半滑舌鳎(Cynoglossus semilaevis)苗种耳石,设置了5个浓度组和3个时间梯度组。结果显示,ALC对半滑舌鳎70–170 d苗种的耳石染色效果明显。综合考虑存活率和标记效果,ALC浸泡标记半滑舌鳎苗种耳石的适宜浓度为100 mg/L(36 h浸泡处理)和150 mg/L(24 h浸泡处理)。相同浸泡时间下,随着ALC浓度的增加,耳石上标记信号强度增加,但实验鱼死亡率升高;相同浸泡浓度下,随着时间的增加,耳石上标记信号强度增加,实验鱼死亡率也会升高。对标记的170 d苗种进行养殖实验,每30 d取样矢耳石进行观察,发现4个月后实验鱼生长良好,且耳石荧光信号仍可清晰检测到,表明ALC在半滑舌鳎矢耳石上形成的荧光标记可长期存续,标记效果好。研究结果可为半滑舌鳎大规模标志放流及早期生活史研究提供群体身份识别技术支持。     

茜素红S对异齿裂腹鱼稚鱼标记效果研究

为探索茜素红S对异齿裂腹鱼稚鱼浸泡标记的适宜条件,采用半静态式水生生物急性毒性试验法确定茜素红S对异齿裂腹鱼稚鱼安全质量浓度,利用不同质量浓度的茜素红S溶液对45日龄的异齿裂腹鱼稚鱼浸泡不同时间,探究各试验组稚鱼死亡率、生长情况、标记效果和标记有效保存时间,确定茜素红S对异齿裂腹鱼稚鱼的适宜标记条件。试验结果表明：茜素红S对异齿裂腹鱼稚鱼24、48、72、96 h半致死质量浓度分别为225.85、147.72、139.21、137.66 mg/L,安全质量浓度为28.99 mg/L;0～125 mg/L的茜素红S溶液浸泡稚鱼24 h,质量浓度为75～125 mg/L稚鱼耳石标记效果可见,当茜素红S质量浓度为125 mg/L时,稚鱼死亡率显著升高(P<0.05);75 mg/L茜素红S浸泡稚鱼12～96 h,浸泡24～96 h稚鱼耳石标记效果可见,当浸泡时间为96 h时,稚鱼死亡率显著升高(P<0.05);在4个月的续养过程中,75 mg/L茜素红S溶液浸泡72～84 h, 4个月后稚鱼耳石标记效果依旧可见。综合生长、存活率、染色效果及标记保存时间,建议采用75 mg/L茜素...     

四环素标记斑马鱼耳石效果及对抗氧化力的影响

为了研究四环素标记鱼类的效果及其对抗氧化能力的影响,设置100、150、250 mg/L浓度梯度的盐酸四环素(TCH)溶液和不同浸泡时间(18 h和24 h)对毒理实验常用对象斑马鱼(Danio rerio)进行浸泡标记,通过观察TCH浸泡标记后30 d内斑马鱼耳石的标记效果以及浸泡后第1、3、9、15 d时鱼体内三种抗氧化酶(SOD、CAT、GSH-Px)的活性变化,综合分析其标记鱼类的效果和安全性。结果显示,在150 mg/L和250 mg/L浓度的TCH溶液中浸泡18 h或24 h,斑马鱼耳石标记率可达100%,且耳石上可见清晰的黄色标记轮纹。浸泡18 h时,中高浓度组(150 mg/L和250 mg/L)斑马鱼的抗氧化酶活性在浸泡后的第3 d显著增加。浸泡24 h时,各浓度组的抗氧化酶活性在第1 d显著升高,250 mg/L浓度组的SOD和GSH-Px活性分别在第1 d和第9 d显著低于对照组,各组的抗氧化酶活性均在第15 d时恢复为对照组水平。结果表明,100 mg/L和150 mg/L浓度的TCH浸泡液对斑马鱼的抗氧化酶活性起诱导作用,250 mg/L浓度的TCH溶液浸泡,对其抗氧化酶活性产生了短期的抑制作用并可能对机体产生可恢复的氧化损伤。     

淡水中心在淡水鱼增殖放流标记技术研究上取得新进展

<正>目前，有效的标记手段和增殖放流评价方法，仍是制约增殖放流应用工作的“瓶颈”问题。淡水渔业研究中心渔业微化学实验室选取鲢鱼和鲫鱼，通过鱼体茜素络合物（ALC）荧光标记和锶元素浸泡标记，探索相关标记方法在鱼类多类硬组织标记和效果评价上的适用性特征和优化途径。研究发现，ALC不仅能够成功标记鲢鱼幼鱼广受关注的耳石（矢耳石、星耳石、微耳石），     

Mass-marking of coregonid larvae and fry by tetracycline tagging of otoliths

Abstract. Larvae and juveniles of two species of Coregouns sp. were immersed in tetracycline (TC) solutions (200–800)mg/1) and fluorescent marks in the otoliths from fish were identified under a UV-light microscope. Whitefish. Coregonus Iavaretus (L.). juveniles (average weight 191 mg) immersed in TC solution retained their marking until the 127th day, hut 40% of the population lost their TC marking after wintering. Whitefish and peled. C. peled Gmelin, larvae retained TC marking from the TC solution (400 mg/1) if water temperature was higher than 5°C, Whitefish juveniles retained clear marking in the otolith after 120 days of post-immersion rearing. Peled larvae immersed in TC solution were grown in a pond and the fluorescent otolith centre of 55·4-g fish was well visible after processing. Nearly 100% of peled larvae grown for 176 days can he identified based on fluoresceent marks observed under a UV-lighl microscope.     

Daily and sub-daily otolith increments of larval and juvenile walleye pollock, Theragra chalcogramma (Pallas), as validated by alizarin complexone experiments

Walleye pollock (Theragra chalcogramma) were reared from eggs to the juvenile life stage to study daily increment formation in the sagittae otoliths, which are routinely used for age and growth analyses. The apparent deposition of sub-daily growth increments becomes problematic for determining fish age from the late larval stage throughout the juvenile (young-of-the-year) development stage. Otolith marking experiments were conducted to determine interpretation criteria to differentiate between daily and sub-daily increments. Immersion of larval and transforming walleye pollock in 25 mg/l of alizarin complexone (ALC) for 6 h once a week produced a fluorescent mark on the day of staining. Evidence of six well defined and equally spaced increments counted between the weekly ALC marks validated the deposition of daily increments. The daily increments gradually increased in width as the fish/otolith grew. The criteria for determining the presence of sub-daily increments between the daily increments were (1) weak optical definition and (2) a sudden change in incremental distance that lasted for one or two increments and were approximately <0.5 μm in width. Growth problems that occurred during the experiments were identified on otoliths as reductions in daily incremental widths and optical definition, which continued for several days. Otoliths from field-collected fish have also shown similar changes in daily increment properties during the juvenile stage, which may be an indicator of an environmental influence. The criteria for defining different increment types help to resolve our current age determination issues for late larval and early juvenile stage walleye pollock from the Gulf of Alaska.     

The use of alizarin complexone for immersion marking of the otoliths of embryos and larvae of the turbot, Scophthalmus maximus (L.): dosage and treatment time

The purpose of this work was to determine the efficacy of marking turbot, Scophthalmus maximus (L.), eggs and larvae with alizarin complexone (ALC) for use in enhancement programmes and in studies of the ecology and behaviour of the species. The main aim was to determine the optimum dosage and treatment time for large batches of larvae destined for release into the sea.Eggs and larvae were immersed in a range of ALC concentrations (0–60 mg L^–1) for various times (3–24 h). On day 8 after hatching, eggs marked with the different ALC dosages had 99% marking success and had mainly good-quality or very-good-quality marks, whereas larvae achieved 100% success and had mainly very-good-quality marks. The best results were obtained after marking larvae with ALC at 60 mg L^–1 for 6–24 h.During the first months after marking, the fluorescent rings formed in the otoliths were easily detected when directly examined with a UV microscope, no prior preparation of the otoliths was necessary, but after 7 months the marks become faint as the otoliths get thicker, so it is necessary to cut or polish the otoliths before examination.     

Validation of daily increment formation in otoliths of immature and adult Japanese anchovy <Emphasis Type="Italic">Engraulis japonicus</Emphasis>

In order to validate daily increment formation in otoliths of immature and adult Japanese anchovy Engraulis japonicus, three rearing experiments using chemical marking of otoliths were conducted on adult anchovy in summer 2004 and immature anchovy in summer 2005 and in winter 2006. In the two experiments conducted in summer, the number of otolith microincrements between alizarin complexone (ALC) marks showed that microincrements were formed daily. In the summer 2005 experiment, immature anchovy under conditions of reduced daily food rations also showed daily microincrement formation. Average increment width was 0.9 μm in adults and 1.8–3.1 μm in immature anchovy. In contrast, no clear increments were observed between ALC marks on the otoliths from the experiment in winter 2006, and scanning electron microscope (SEM) observations failed to confirm clear increment formation. We consider that low water temperatures (<13–14°C) restricted otolith growth and lowered the contrast between the discontinuous and the incremental zones of the otolith increments. For age estimation of Japanese anchovy, clear increments wider than about 1 μm in the otolith can be regarded as daily increments. However, daily age estimation of immature and adult anchovy that experience low water temperatures in winter may be difficult due to the obscurity of the increments.     

Otolith microstructure of brown sole <Emphasis Type="Italic">Pseudopleuronectes herzensteini</Emphasis>: validation of daily ring formation and the occurrence of microstructure denoting metamorphosis

Brown sole Pseudopleuronectes herzensteini larvae and juveniles were reared to validate daily otolith ring formation. At 15°C, a check (a distinct ring) formed on the sagittae and lapilli at 6 days after hatching, and clear increments regularly formed outside the check. For both otoliths, the relationship between the number of days after hatching and number of increments was linear, and the slope of the line was approximately 1; therefore, daily formation was validated. At 12°C, the check formed on the lapillus 8 days after hatching. Accessory primordia (AP) began forming on the sagittae of metamorphosing larvae, and the shape of the sagittae became complicated. AP were not formed on the lapillus; concentric rings were formed throughout larval and juvenile stages. Wide and obscure increments formed on the lapilli during metamorphosis (metamorphosing zone, MZ). Based on MZ, concentric rings that have formed on the lapilli of juveniles can be separated into larval and juvenile rings. The morphs of large juveniles’ lapilli were bilaterally asymmetric, and the blind-side lapilli were most suitable for otolith microstructure analysis. This study provides fundamental information for otolith microstructure analysis in wild brown sole.     

Entrainment and advection of larval sardine,Sardinops sagax,by the East Australian Current and retention in the western Tasman Front

The poleward flowing East Australian Current (EAC) drives sporadic upwelling, entrains coastal water and forms the western Tasman Front (wTF), creating a mosaic of water types and larval transport routes along south eastern Australia. The spatial distribution, otolith chemistry and growth rates of larval sardine (Sardinops sagax) were examined to infer spawning location and larval transport. A gradient of increasing larval size from north to south along the shelf was not detected but was evident between the shelf and offshore in the wTF. Here larvae were larger and older. Based on the occurrence of newly hatched larvae, spawning by S. sagax between southern Queensland and mid New South Wales (NSW) was more extensive than previously reported. The otolith chemistry from two wTF larval size classes differed, implying different origins. The otolith chemistry of wTF post‐flexion larvae was similar to larvae from northern NSW, whereas wTF flexion larvae were similar to larvae observed nearby from mid‐NSW. Two possible larval transport routes, direct and indirect, are inferred from otolith chemistry, current velocities and a previously published particle tracking study. Either larvae from northern NSW were advected south and entrained with younger larvae directly into the wTF, or larvae from a range of shelf regions were advected around the southern edge of an anticyclonic eddy, to join younger larvae directly entrained into the wTF. Based on the co‐occurrence of larval ages and sizes in the wTF and their advection routes, the wTF appears to be an important larval retention zone.