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慢性暴露于亚致死浓度铜下的尼罗罗非鱼:对生长、抗氧化、非酶抗氧化、氧化应激和非特异性免疫反应的影响。

Chronic exposure of Oreochromis niloticus to sub-lethal copper concentrations: Effects on growth, antioxidant, non-enzymatic antioxidant, oxidative stress and non-specific immune responses.

机构信息

Biomaterials and Biotechnology in Animal Health Lab, Department of Animal Health and Management, Alagappa University, Karaikudi, 630004, Tamil Nadu, India.

Foundation for Innovative Research in Science and Technology, Kelavannanvilai, NGO Colony Road, Nagercoil, Tamil Nadu, India.

出版信息

J Trace Elem Med Biol. 2019 Sep;55:170-179. doi: 10.1016/j.jtemb.2019.06.011. Epub 2019 Jun 27.

Abstract

BACKGROUND

Trace elements of copper (Cu) are one of the main forms of ecological noxious waste in freshwater systems that affect the survival and development of organisms. The objective of the current study was to investigate the effects of chronic exposure to Cu on the growth, oxidative stress, immune and biochemical response in the Nile tilapia, Oreochromis niloticus.

METHODS

Three groups of O. niloticus were tested as follows; the first group was used as the control (not treated with Cu in water), while the 2nd and 3rd groups were exposed to (low) 40 μg L and (high) 400 μg L concentrations of Cu added to water, respectively. The duration of the experiment, which was conducted in triplicate, was 60 d. End points were evaluated on days 30 and 60. Following 30 d and 60 d of exposure to Cu, the fish were removed from experimental tanks to determine growth. Consequently, blood samples were collected from caudal veins at the end of the trial period (30 d and 60 d) and serum was separated to evaluate different immunological parameters, such as lysozymes (LYZ), respiratory burst activity (RBA) and myeloperoxidase (MPO). Gill and liver tissues were collected for evaluation of Cu and certain biochemical parameters as follows: antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST); non-enzymatic antioxidants such as glutathione (GSH) and metallothionein (MT), and oxidative stress indicators such as malondialdehyde (MDA) and protein carbonyl (PCO). The results pertaining to treatments and the control were compared using two-way ANOVA and Tukey's HSD test. The level of significance was set at P ≤ 0.05. Data were expressed as mean ± SD.

RESULTS

Chronic exposure to Cu did not induce any mortality in fish during the test period. However, following exposure to Cu, growth of fish in the exposed groups was affected more than that in the control group (unexposed to Cu). In addition, accumulation of Cu in the liver tissue was higher than that in the gill tissues of fish exposed to Cu, compared to that in the control. Gill and liver tissues of Cu-exposed fish showed a significant (P ≤ 0.05) reduction in the activities of the antioxidant enzymes, SOD, CAT, GPx, and GST, compared to those of unexposed fish. Non-enzymatic antioxidants, GSH and MT, in gill and liver tissues were significantly increased (P ≤ 0.05) in fish exposed to both concentrations of Cu, compared to those in unexposed fish. Oxidative stress indicators, MDA and PCO in gills and liver of Cu-exposed fish was significantly (P ≤ 0.05) at both tested concentrations, when compared to control group. Non-specific immune response of LYZ, RBA, and MPO activity in serum decreased significantly (P ≤ 0.05) in Cu-exposed fish, compared with that of unexposed fish.

CONCLUSION

Overall, the present results highlighted that chronic exposure to Cu ions may exert a strong effect on the antioxidant and immune responses of O. niloticus. Changes in antioxidant enzymes, oxidative stress effects and immune parameters during post-chronic metal exposure may indicate the potential of these parameters as biomarkers of metal toxicity in aquatic ecosystems.

摘要

背景

铜(Cu)微量元素是淡水系统中主要的生态有毒废物形式之一,会影响生物的生存和发展。本研究的目的是研究慢性暴露于 Cu 对尼罗罗非鱼 Oreochromis niloticus 的生长、氧化应激、免疫和生化反应的影响。

方法

将三组 O. niloticus 进行如下测试;第一组作为对照组(水中不添加 Cu),而第二组和第三组分别暴露于(低)40μg/L 和(高)400μg/L 的 Cu 浓度。实验持续 60 天,进行了三倍重复。在 30 天和 60 天暴露于 Cu 后,将鱼从实验罐中取出以评估生长。随后,在试验结束时(30 天和 60 天)从尾静脉采集血液样本,并分离血清以评估不同的免疫参数,如溶菌酶(LYZ)、呼吸爆发活性(RBA)和髓过氧化物酶(MPO)。收集鳃和肝脏组织以评估 Cu 和以下生化参数:抗氧化酶,如超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)和谷胱甘肽-S-转移酶(GST);非酶抗氧化剂,如谷胱甘肽(GSH)和金属硫蛋白(MT),以及氧化应激指标,如丙二醛(MDA)和蛋白质羰基(PCO)。使用双因素方差分析和 Tukey 的 HSD 检验比较处理组和对照组的结果。显著性水平设定为 P ≤ 0.05。数据表示为平均值±标准差。

结果

在测试期间,慢性暴露于 Cu 并未导致鱼类死亡。然而,暴露于 Cu 后,暴露组的鱼的生长受到的影响大于未暴露于 Cu 的对照组。此外,与对照组相比,暴露于 Cu 的鱼肝脏组织中 Cu 的积累量高于鳃组织。暴露于 Cu 的鱼的鳃和肝脏组织中的抗氧化酶 SOD、CAT、GPx 和 GST 的活性显著降低(P ≤ 0.05)。与未暴露于 Cu 的鱼相比,暴露于 Cu 的鱼的鳃和肝脏组织中的非酶抗氧化剂 GSH 和 MT 显著增加(P ≤ 0.05)。暴露于 Cu 的鱼的鳃和肝脏中的氧化应激指标 MDA 和 PCO 在两个测试浓度下均显著升高(P ≤ 0.05),与对照组相比。暴露于 Cu 的鱼的血清中非特异性免疫应答 LYZ、RBA 和 MPO 活性显著降低(P ≤ 0.05)。

结论

总的来说,本研究结果强调了慢性暴露于 Cu 离子可能对 O. niloticus 的抗氧化和免疫反应产生强烈影响。在慢性金属暴露后,抗氧化酶、氧化应激效应和免疫参数的变化可能表明这些参数作为水生生态系统金属毒性的生物标志物的潜力。

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