茜素红S对中华倒刺鲃幼鱼不同组织抗氧化酶活性和脂质过氧化的影响

以中华倒刺鲃幼鱼[平均体长（12.3±0.6）cm,平均体重（41.8±3.6）g]为研究对象,通过测定不同浓度茜素红S溶液（ARS）浸泡24h后肝脏、脑和鳃组织中抗氧化酶（SOD、CAT、GSH-Px）的活性和丙二醛（MDA）含量,评估ARS对中华倒刺鲃幼鱼的生理生化影响。结果表明:除脑组织GSH-Px随着ARS浓度的升高而不断升高外,肝脏、鳃和脑组织的抗氧化酶活性均表现为低浓度被诱导而高浓度受抑制的规律,与ARS浓度呈抛物线型剂量效应关系。肝脏、鳃、脑组织抗氧化酶活性达到最大值所对应的浓度分别是300、300和400 mg/L。除鳃部MDA含量随着茜素红S浓度升高不断升高外,脑和肝脏组织的丙二醛含量随着ARS浓度的升高呈现出先降低后上升趋势。根据试验的结果,推荐120-200 mg/L为ARS染色标记中华倒刺鲃幼鱼（体长10 cm）的适宜浓度。     

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.     

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。     

力竭追赶训练对两种鲤科鱼类呼吸循环系统参数和力竭运动后代谢特征的影响

为了考察力竭追赶训练对中华倒刺鲃(Spinibarbus sinensis)和岩原鲤(Procypris rabaudi)幼鱼呼吸循环系统和力竭运动后代谢特征的影响, 在(25±0.5)℃条件下, 将2种实验鱼各60尾 [体重分别为(28.36±0.08) g和(19.53±0.13) g]随机等分成对照组和训练组, 训练组进行1次/d共21d的力竭追赶训练。随后测定各组实验鱼的心脏和鳃指数、血液指标以及力竭运动后过量耗氧(Excess post-exercise oxygen consumption, EPOC)。结果发现: 2种鱼心脏指数、鳃指数、血红蛋白含量和红细胞数目在各自对照组和训练组之间都没有显著性差异; 岩原鲤对照组的鳃指数显著低于中华倒刺鲃对照组(P<0.05), 但心脏指数、血红蛋白含量和红细胞数目与中华倒刺鲃对照组无显著性差异。中华倒刺鲃训练组的运动前代谢率、运动代谢峰值、峰值比率、代谢恢复速率与其对照组没有显著差异, 但运动后恢复时间和过量耗氧显著大于对照组(P<0.05); 岩原鲤训练组的运动前代谢率显著低于其对照组(P<0.05), 但运动代谢峰值、峰值比率、运动后恢复时间、过量耗氧、代谢恢复速率与其对照组没有显著性差异。岩原鲤对照组的运动前代谢率、运动代谢峰值、过量耗氧和代谢恢复速率分别显著低于中华倒刺鲃对照组(P<0.05), 但峰值比率和运动后恢复时间与中华倒刺鲃对照组无显著性差异。研究表明: (1)力竭追赶训练对中华倒刺鲃和岩原鲤幼鱼的呼吸和循环系统参数没有产生显著性影响; (2)力竭追赶训练显著提高了中华倒刺鲃幼鱼的无氧代谢能力; (3)中华倒刺鲃幼鱼力竭运动前后代谢率显著高于岩原鲤幼鱼, 这可能与其较为活跃的习性有关。     

溶氧水平对鳊鱼、中华倒刺鲃幼鱼游泳能力的影响

在(25 1)℃条件下, 以鳊鱼(Parabramis pekinensis)体重(4.70 0.11)g, n=32、中华倒刺鲃(Spinibarbus sinensis)体重(3.26 0.06)g, n=32幼鱼为研究对象, 采用鱼类游泳代谢测定仪在水体溶氧为8、4、2、1 mgO2/L条件下分别测定其临界游泳速度(Ucrit)和游泳代谢率(MO2), 并计算出静止代谢率(MO2rest)、最大游泳代谢率(MO2max)、代谢范围(MS)及单位位移能耗(COT)等相关参数。结果显示, 随着溶氧水平的下降, 鳊鱼、中华倒刺鲃幼鱼的Ucrit均逐渐下降, 除中华倒刺鲃幼鱼的值在4与8 mgO2/L下没有显著性差异外, 其他各组均差异显著(P0.05); 在同一溶氧水平下的中华倒刺鲃Ucrit显著大于鳊鱼(P0.05)。两种鱼的MO2max和MS均随DO的下降而显著下降, 但MO2rest在溶氧水平低于1 mgO2/L才显著下降(P0.05)。研究还发现鳊鱼、中华倒刺鲃幼鱼的MO2在同一游泳速度下随溶氧水平下降而降低, 而在相同溶氧水平下随游泳速度的上升而显著升高(P0.05); COT随游泳速度上升而显著降低(P0.05), 但在高游泳速度下相对稳定, 在同一游泳速度下随着DO的下降有所减小。中华倒刺鲃的COT整体上小于鳊鱼, 且在低游泳速度下差异更大。87.5%(1-8 mgO2/L)溶氧水平的下降导致两种鱼类相似的Ucrit变化(53% vs. 50%), 但溶氧水平由8降到4 mgO2/L时, 鳊鱼32%MO2max的下降导致Ucrit下降13%, 但同样的溶氧水平下降虽然导致中华倒刺鲃的MO2max下降20%, 但由于MO2rest和COT的下降, 其Ucrit并没有显著的变化。实验结果表明: 不同溶氧水平对不同鱼种游泳能力的影响存在差异, 这种差异与其代谢对策密切相关。       

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。     

茜素络合物对唐鱼耳石标记效果以及生长和存活率的影响

用茜素络合物(ALC)对唐鱼(Tanichthys albonubes)进行荧光标记,通过测定唐鱼耳石的标记率、死亡率、SGR和DWG等指标,讨论不同浓度ALC对唐鱼耳石标记效果和对其生长和存活的影响,探讨应用该标记技术追踪唐鱼野外种群迁移行为的可行性。结果表明:在温度28~30℃,浓度50 mg·L-1、80 mg·L-1茜素络合物溶液中浸泡24 h,唐鱼仔鱼、稚鱼死亡率为0,其中80 mg·L-1浓度处理组耳石荧光标记环检出率为100%,而在100mg·L-1浓度下,仔、稚鱼死亡率均达到80%;幼鱼在浓度为80、100 mg·L-1条件下无死亡,其中100 mg·L-1浓度处理组耳石标记环检出率为100%,而150 mg·L-1浓度下幼鱼死亡率为44%;成鱼在100、150 mg·L-1浓度下无死亡,且耳石标记率均为100%,而在200mg·L-1浓度下成鱼死亡率达到100%;此外,稚鱼在80 mg·L-1最适浓度下浸泡24 h后继续饲养90 d,标记组和对照组死亡率均为0,两组体质量、SGR和DWG差异不显著;唐鱼仔稚鱼、幼鱼和成鱼耳石标记的最适茜素络合物浓度分别为80、100和150 mg·L-1。     

饥饿对中华倒刺鲃幼鱼代谢、个性和集群的影响

在自然界中,环境变化、季节更替和人为因素造成食物资源时空分布的不均一性,导致鱼类经常面临食物资源短缺的环境胁迫,对其能量代谢和行为造成一定影响。为考察食物资源短缺下暖水性鲤科鱼类能量代谢、个性与集群行为的应对策略及其可能的内在关联,选取中华倒刺鲃(Spinibarbus sinensis)幼鱼为实验对象,分别测定饥饿组(2周)和对照组(维持日粮)在处理前后实验鱼的标准代谢率(Standard metabolic rate,SMR)、个性行为(勇敢性、探索性和活跃性)以及实验处理后的集群行为(凝聚力和协调性)。研究发现:(1)饥饿组和对照组实验过程中实验鱼SMR均显著下降,但仅饥饿组实验鱼SMR具有重复性;(2)饥饿导致中华倒刺鲃幼鱼勇敢性、探索性、活跃性均显著增加;(3)饥饿导致群体成员间距离缩短,游泳速度及其同步性上升。研究表明:饥饿后的中华倒刺鲃不仅适应性降低SMR以减少能量消耗,而且呈现出更高的勇敢性、探索性和活跃性以利于获取食物资源;饥饿迫使中华倒刺鲃群体提高凝聚力和协调性,可能有助于提高群体的生存能力。     

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)。     

Experimental evaluation of calcein and alizarin red S for immersion marking of silver carp Hypophthalmichthys molitrix (Valenciennes, 1844)

Calcein (CAL) and alizarin red S (ARS) at concentrations of 50–200 and 150–300 mg/L, respectively, were used for immersion marking of juvenile silver carp Hypophthalmichthys molitrix (79.65 ± 2.11 mm total length, mean ± SD). The marked fish were kept in separate tanks (0.012 individuals per litre, rearing temperature 18.4–25.7°C) for 360 days. Each experimental treatment group consisted of three replicates (16 individuals per replicate). Immersion for 24 h produced detectable marks in the sagittae, lateral line and non‐lateral line scales, and fin rays (dorsal, pectoral, ventral, anal, and caudal) at 360 days post‐marking. Acceptable marks in the sagittae were observed for CAL at concentrations of 150–200 mg/L and for ARS at concentrations of 200–300 mg/L. Fluorescent marks were poorly visible in all non‐lateral line scales from both the CAL‐ and ARS‐treated groups. Acceptable fluorescent marks in the lateral line scales and fin rays were detected for CAL at concentrations of 150–200 and 100–200 mg/L, respectively, and for ARS at concentrations of 200–300 and 150–300 mg/L, respectively. In particular, optimal marks were observed at the highest concentrations investigated in lateral line scales (200 mg/L CAL, 300 mg/L ARS) and fin rays (200 mg/L CAL, 200–300 mg/L ARS). There was no significant difference in the survival or growth of marked fish compared to controls throughout the experiment (P > 0.05).     

Experimental evaluation of using calcein and alizarin red S for immersion marking of bighead carp Aristichthys nobilis (Richardson, 1845) to assess growth and identification of marks in otoliths,scales and fin rays

In order to evaluate the effects of immersion marking with calcein (CAL) and alizarin red S (ARS) on growth and mortality of juvenile bighead carp Aristichthys nobilis, and assess mark quality in otoliths, scales, and fin rays, CAL from 50 to 200 mg L^?1 and ARS from 150 to 300 mg L^?1 concentrations were used. With the exception of non‐lateral line scales from 50 mg L^?1 CAL treatments, immersion for 24 h produced detectable marks in sagittae, lateral line and non‐lateral line scales, and fin rays (dorsal, pectoral, ventral, anal, and caudal) at 100 days post‐marking. Detectable fluorescent marks in sagittae were readily observed at concentrations of 150–200 mg L^?1 CAL or 150–300 mg L^?1 ARS. Marks were poorly visible in all non‐lateral line scales from both CAL‐ and ARS‐treated groups. Fluorescent marks were readily detected in lateral line scales at 100–200 mg L^?1 CAL or 150–300 mg L^?1 ARS, and in fin rays at 150–200 mg L^?1 CAL or 150–300 mg L^?1 ARS. In particular, optimal marks were observed at the highest concentrations investigated in sagittae (300 mg L^?1 ARS), lateral line scales (150–200 mg L^?1 CAL or 250–300 mg L^?1 ARS), and fin rays (200 mg L^?1 CAL or 250–300 mg L^?1 ARS). However, fluorescent marks visible to the naked eye were not produced by any of the CAL or ARS treatments in sagittae, scales, or fin rays during this experiment. In addition, there was no significant difference on survival and growth of marked fish compared to controls throughout the experiment (P > 0.05).     

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。     

Marking otoliths of brown trout (Salmo trutta L.) embryos with alizarin red S

In this study a time‐ and cost effective method for marking early life stages of brown trout (Salmo trutta L.) with alizarin red S (ARS) is presented. The main intention was the development of a protocol to mark embryos. When stocking eyed‐eggs, later recognition depends on an adequate tagging method featuring reasonable expenditures, low mortality and high marking success. Best results were obtained using ARS concentrations of 500 mg/l. Consistently clearly visible marks could be detected under a fluorescence‐stereomicroscope equipped with a dsRed filter, without processing the otoliths.     

Mass-marking of brown trout (Salmo trutta L.) larvae by alizarin: method and evaluation of stocking

This study describes otolith marking of brown trout (Salmo trutta L.) larvae by immersion in different solutions of alizarin red S (ARS). The best results were obtained after marking with ARS at a concentration of 150 mg L^?1. To evaluate the efficiency of stocking with brown trout fry, 10 000 20‐day‐old larvae were marked in years 2002 and 2003 with ARS and released 2 weeks later into sections of a river with natural brown trout reproduction. Electro‐fishing surveys carried out 2 months after stocking in 2002 revealed that only 4.8% of all caught young‐of‐the‐year trout originated from stocking; in 2003 the percentage was 8.9%. Based on the substantial natural reproduction and the low ratio of stocked to wild trout, it was recommended to discontinue stocking.     

Optimizing the mass marking of fish with alizarin red S: an example with glass eels

Fish marking is an essential tool for fisheries management, especially for evaluating the stocking of endangered fish species to support conservation and sustainable use of fish stocks. Batch marking of young European eels Anguilla anguilla (L.) prior to stocking is recommended as the benefits of stocking for the spawning stock can be evaluated by recapturing marked fish over time, therefore mass marking of young eels with substances such as alizarin red S (ARS) is becoming increasingly important. To improve the marking method and reduce marking costs when immersing glass eels in an ARS solution, eight laboratory experiments under varying conditions (e.g., temperature, ARS concentration, immersion time, osmotic induction, fish density) and with ARS from different suppliers were carried out. The results show that optimal marking of glass eels can be carried out in the field or during transport by putting approximately 50 g of glass eels per liter in 150 mg L⁻¹ ARS solution for 3 h at 10–15°C. Lower concentrations did not result in reliable marking. Water temperatures of 5°C and below can have a stunning effect on the eels and increase mortality significantly, regardless of the concentration of ARS. Glass eel densities below 50 g L⁻¹ in the marking bath increase marking costs unnecessarily, while a higher density of 100 g L⁻¹ resulted in significantly higher mortality and lower marking success. A somewhat more difficult but less expensive alternative is to bathe the fish in a saline solution of 1% (10 PSU) of 80 mg L⁻¹ ARS for 3 h at 10°C. Costs can also be significantly reduced by choice of supplier for ARS, but care should be taken as the quality of the powder appears to vary (mean percentage of sufficiently marked eels ranged from 59% to 91% among suppliers in the present study) and can lead to marking failure. The optimal marking conditions can help ensure that stocked glass eels can be reliably identified in future studies to assess stocking benefits while reducing costs.     

青海湖裸鲤不同年龄鉴定材料的年轮特征

通过各种年龄鉴定材料比较研究了青海湖裸鲤的年轮特征。微耳石和星耳石可用于鉴定年龄,而矢耳石易碎,不适合鉴定年龄。4种年龄鉴定材料对青海湖裸鲤年龄的判别能力为:背鳍条>微耳石>臀鳞>脊椎骨。对8龄以下个体,用背鳍条磨片鉴定年龄效果较佳;对8龄以上个体,臀鳞、背鳍条磨片和脊椎骨上的年轮计数明显低于微耳石磨片,微耳石磨片是高龄青海湖裸鲤较为可靠的年龄鉴定材料。