| Spinibarbus sinensis belongs to Spinibarbus, Barbinae, Cyprinidae, Cypriniformes.To explore a method which is suitable for the releasing fish is is an important prerequisite about the effect of monitoring and evaluation of artificial enhancement. At present,immersing fish in fluorescent dyes is the most prefectest method. Now domestic studies on immersing fish with fluorescent dyes are focusing on the choice of fluorescent pigments and immersion concentrations, and immersion time is always set at 24 h.However, there is little known about the effect of marking with different immersion concentrations, different size of fish and the impact degree caused by the stress when immersing fish in fluorescent dyes. In present study, we used the juvenile Spinibarbus sinensis as experiment model to research the toxicity of alizarin red S to the juvenile Spinibarbus sinensis. Then the antioxidant enzymes and blood biochemical index were studied after immersing 24 h, transferring into freshwater 5 d, 10 d to assess the toxicity of alizarin red S to the juvenile Spinibarbus sinensis and to test its security. At the same time, exploring the resistance-responsive caused by alizarin red S to know the effect of the physiology and biochemistry. Then based on the dyeing effect of olotith and scales to find out the suitable marking concentration and immersion time for juvenile Spinibarbus sinensis. These results could provide theoretical basis and reference for marking techniques in living aquatic resources enhancement. The main results were as follows:1. The results showed that the 24 h,48 h,72 h,96 h median lethal dose and safe dose of alizarin red S for juvenile Spinibarbus sinensis were 788.3、744.5、653.5、595.6 and59.56 mg/L, it showed that the toxicity of alizarin red S were low for juvenile Spinibarbus sinensis, and it were feasible to mark releasing activity.2. The stained mark were red which soaked in alizarin red S, and the best results should be observed under green excitation light. The mark efficiency and quality of otoliths and scales increased with alizarin red S concentration rising. In addition to 100mg/L concentration group, other concentrations otolith mark rate reached 91.7%. The effect of marking were not ideal in 100 mg/L, 200 mg/L concentration group; when alizarin red S concentrations were more than 300 mg/L, good quality mark(n ≥ 2)percentage of asteriscus and lapillus were respectively over 95.9% and 84.5%, the effect of marking were ideal. Meanwhile observeing the dyeing effect of scales found: the effectof marking were not ideal in all of concentration groups. The dyeing quality of asteriscus were better than lapillus, the dyeing quality of otolith were better than scales.3. The research of blood biochemical parameters showed that: after alizarin red S soaking 24 h, the levels of TP, GLB, CRE in plasma increased with the alizarin red S concentration riseing, when concentration of alizarin red S were more than 300 mg/L,there was extremely significant difference(p < 0.01) between experimental group and control group. With the alizarin red S increaseing the concentration of ALB, ALP, GLU showed increased firstly and then decreased, the peak valuer were in 300 mg/L, 200 mg/L,300 mg/L espectively, the concentration of AST, ALT increased firstly and then decreased with ARS concentration increaseing, the minimum were both in the 100 mg/L concentration group.Temporary culture 5 d later, The change trend of TP, ALB, GLB, T-CHO, ALP,CRE in juvenile plasma were basically consistent, when concentration of alizarin red S were more than 300 mg/L, the 6 indicators decreased with concentration of alizarin red S increasing, and the value in 400 mg/L, 500 mg/L experimental group were significantly lower than the control group(p < 0.05); only the value of GLU in 500 mg/L concentration group was significantly lower than the control group; The change trend of AST, ALT in juvenile plasma were consistent, when concentration of alizarin red S were more than 300mg/L, the value of transaminase increased constantly with concentration of alizarin red S increasing, the value of transaminase in 400 mg/L, 500 mg/L concentration groups was significantly higher than the control group(p < 0.05). Temporary culture 10 days later, in addition to the biochemical index in 500mg/L concentration group were significant differences with control group(p < 0.05), there were no difference between other concentrations with control group.The result combined with 3 times data of blood biochemical parameters showed: when the dyeing concentration of alizarin red S were lower than 200 mg/L, it was safe for juvenile S. sinensis; when the dyeing concentration of alizarin red S were equal to and higher than 300 mg/L, although juvenile S. sinensis can not be dead, but it would cause damage to gill, liver, kidneys and other organs in a certain degree. The tissue damage which were coused in 300 mg/L, 400 mg/L concentration can be restored after temporary culture of 5 d; The tissue damage which were coused in 500 mg/L concentration can not be restored after temporary culture of 10 d, so the concentration of alizarin red S can not exceed 400 mg/L during soaking juvenile S. sinensis.4. Through researching the activity of antioxidant enzymes(CAT, SOD, GSH-Px) of different tissues and change of malondialdehyde(MDA) content in different concentration of alizarin red S, it can be evaluation the effect of alizarin red S on he physiological and biochemical for juvenile spinibarbus sinensis. The results showed that under the stress of different concentration of alizarin red S, in addition to GSH-Px activity of brain tissue increased with concentration of alizarin red S rising, antioxidant enzyme activity of brain, liver and gill in the test groups present the same rule that low concentrations were induced and high concentration of inhibition. It present a dose effect of parabolic type relationship between antioxidant enzyme activity and concentration of alizarin red S. The concentration which liver, gill, brain reached the maximum of antioxidant enzyme activity corresponded respectively were 300 mg/L, 300 mg/L, 400mg/L. After temporary culture 5 d, 3 kinds of antioxidase activity of the brain tissue still showed a rising trend with the increasing of alizarin red S concentration; but 3 kinds of antioxidase activity of liver and gill tissue were firstly increased and then decreased,antioxidant enzyme activity in 400 mg/L concentration group was the highest and antioxidant enzyme activity in 500 mg/L group was the lowest. Temporary culture 10 d later, only antioxidant enzymes of different tissues in 500 mg/L concentration group had significant difference with control group(p < 0.05).After 24 h immersion, except that the MDA content of the gills were continuously rising with alizarin red S concentration increasing. the MDA content of the brain and liver tissue decreased firstly and then increased with alizarin red S concentration increasing,After temporary culture 5 d, The content of MDA in brain tissue firstly decreased and then increased with the increasing of alizarin red S concentration, The MDA content of400 mg/L concentration was the lowest(p < 0.05), The MDA content of 500 mg/L concentration was the highest(p < 0.05); The content of MDA in gill constantly creased with increasing of alizarin red S concentration, The content of MDA in more than 300mg/L concentration groups was significant higher than control group(p < 0.05); The content of MDA in liver tissue firstly decreased and then increased with increasing of alizarin red S concentration, The content of MDA in 500 mg/L concentration group was the highest, significantly higher than the control group(p < 0.05). Temporary culture 10 dlater, only the content of MDA in 500 mg/L concentrations group in liver and gill tissue were significantly higher than control group(p < 0.05). Research showed that: the antioxidant enzyme system in different tissues had tissue specificity to alizarin red S stress, Alizarin red S in too high concentrations can cause oxidative damage in different tissues of juvenile spinibarbus sinensis. The gill tissue were the highest sensitivity to alizarin red S stress. So we should consider the sensitivity of gill tissue in the process of fluorescence soaking and to maintain adequate oxygen.when we selected the appropriate concentration of alizarin red S to dye. |
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