慢性氨氮胁迫对幼鲫肝、肾组织结构及非特异性免疫指标的影响

张月; 郝玲; 胡雅菲; 金珂; 杨慧轩; 宋晶

doi:10.19303/j.issn.1008-0384.2024.06.001

慢性氨氮胁迫对幼鲫肝、肾组织结构及非特异性免疫指标的影响

doi: 10.19303/j.issn.1008-0384.2024.06.001

山西农业大学动物科学学院，山西　晋中　030801

基金项目: 山西省“1331工程”重点学科建设计划项目（J201911306）

详细信息

作者简介:
张月（1996 —），女，硕士，主要从事动物环境控制研究，E-mail：1620674729@qq.com

通讯作者:
宋晶（1983 —），男，博士，副教授，主要从事鱼类神经生理和行为研究，E-mail：songjingoak@163.com

中图分类号: S965.117
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出版历程
- 收稿日期: 2024-01-17
- 修回日期: 2024-04-27
- 网络出版日期: 2024-08-15
- 刊出日期: 2024-06-28

Effects of Prolonged Ammonia Nitrogen Stress on Liver and Kidney Histology and Non-specific Immunity of Juvenile Carassius auratus

College of Animal Science, Shanxi Agricultural University, Jinzhong, Shanxi　030801, China

摘要

摘要: 目的研究慢性氨氮胁迫对幼鲫（Carassius auratus）肝、肾组织结构及非特异性免疫指标的影响，为研究慢性氨氮对幼鲫的危害及其集约化养殖水质管理提供理论依据。方法以体重（3.10±0.15）g的幼鲫（C. auratus）为试验对象，通过急性毒性试验，得出96 h半致死质量浓度（LC₅₀）和安全质量浓度（safe concentration, SC），并以此为基础，设0 mg·L⁻¹（对照，CK）、6 mg·L⁻¹（低质量浓度，L组）、15 mg·L⁻¹（中质量浓度，M组）和24 mg·L⁻¹（高质量浓度，H组）4个不同氨氮浓度处理组，试验为期60 d。分别对胁迫第15 、30 、45、60天时的幼鲫肝、肾组织的病理变化特征进行分析，并测定谷丙转氨酶（alanine aminotransferase, ALT）、谷草转氨酶（aspartate aminotransferase, AST）、过氧化氢酶（catalase, CAT）、超氧化物歧化酶（superoxide dismutase, SOD）活性变化。结果氨氮对幼鲫的96 h半致死质量浓度LC₅₀为289.29 mg·L⁻¹，安全质量浓度SC为28.9 mg·L⁻¹。在氨氮胁迫下，幼鲫组织的病理变化主要为肝细胞空泡化，核仁溶解，肝细胞轮廓模糊、排列混乱；肾小管上皮细胞水肿变性，肾小管管腔缩小，肾小球萎缩。在整个氨氮胁迫期间，肝、肾组织中各氨氮处理组的ALT、AST活性呈升高趋势，其中除15 d时L组和M组的ALT在肝脏中与对照组差异不显著（P＞0.05）外，其他各时间段3个胁迫组ALT、AST活性在肝、肾组织中均与对照组差异显著（P＜0.05）。肝脏中CAT、SOD比活力呈先升高后降低趋势，各胁迫组CAT比活力在45 d时与对照组差异显著，SOD比活力在15 d时与对照组差异显著。肾组织中CAT比活力在15 d时与对照组相比显著升高；各胁迫组SOD比活力在各时间段均显著高于对照组。结论氨氮胁迫会导致幼鲫肝、肾组织严重损伤，并伤害其代谢、解毒能力和非特异性免疫能力。在集约化养殖过程中，应及时关注养殖水体中的氨氮含量，减少氨氮胁迫对鲫鱼造成的伤害，提高经济效益。
- 氨氮胁迫 /
- 幼鲫 /
- 肝脏 /
- 肾脏 /
- 非特异性免疫指标
Abstract: Objective Chronic ammonia nitrogen stress on juvenile crucians were studied with observations on liver and kidney histology and non-specific immunity of the fish for an improved aquaculture water quality control. Method Juvenile Carassius auratus with a body mass of (3.10±0.15) g were raised in an environmentally controlled pond. By an acute toxicity test on the fish, the semi-lethal (LC₅₀) and safe concentrations (SC) of ammonia nitrogen over 96 h were determined. Accordingly, a control at 0 mg·L⁻¹ and three treatments groups (i.e., low concentration L at 6 mg·L⁻¹, medium concentration M at 15 mg·L⁻¹, and high concentration H at 24 mg·L⁻¹) of ammonia nitrogen in water were applied to the pond with the juvenile fish for 60 d. Histopathological changes induced by the treatments in the liver and kidney as well as the alternations occurred to the activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), catalase (CAT), and superoxide dismutase (SOD) of the fish were monitored continuously on the 15^th, 30^th, 45^th, and 60^th days. Result The acute ammonia nitrogen toxicity test on the fish showed the LC₅₀ of 289.29 mg·L⁻¹ and SC of 28.9 mg·L⁻¹. The microscopic examination revealed the increasing ammonia nitrogen concentration and stress duration caused symptoms in the fish included blurred and disorganized hepatocytes with some vacuolated and cell nuclei dissolved in hepatic tissue, tumefacient renal tubular epithelial cells, narrowed renal tubular lumen, and atrophied glomerulus in nephridial tissue. The ALT and AST activities in the liver and the kidney rose with increasing ammonia nitrogen in water. The ALT in the livers of fish in L and M groups did not significantly differ from control on the 15^th day (P＞0.05). On the other hand, the ALT and AST in livers and kidneys of all 3 treatment groups were significantly different from control at all testing times (P＜0.05). The specific activity of CAT and SOD in liver of the fish increased initially and decreased after a period of time of the treatments; that of CAT in treatment groups significantly differed from control on the 45^th day; that of SOD significantly differed on the 15^th day, and that of CAT in renal tissue significantly higher than that in control group on the same day; while that of SOD in all treatment groups significantly higher than that in control at all time periods. Conclusion The stress of prolonged high concentrations of ammonia nitrogen in aquacultural water caused severe damage to the liver and kidney tissues in juvenile crucians. The histopathological injuries could result in malfunctioned metabolism, compromised detoxification capacity, and reduced non-specific immunity of the fish. Consequently, for a healthy and productive aquaculture, it was deemed imperative the ammonia nitrogen content in the water be continuously monitored and strictly controlled.
- ammonia nitrogen stress /
- juvenile crucians /
- liver /
- kidney /
- non-specific immunity indices

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图 1 慢性氨氮胁迫对幼鲫肝组织显微结构的影响

Hc：肝细胞；HS：肝血窦；CV：肝细胞空泡化；Bc：细胞轮廓模糊。

Figure 1. Effect of chronic ammonia nitrogen stress on microstructure of liver tissue of juvenile C. auratus

Hc: hepatocytes; HS: hepatic sinuses; CV: vacuolization of hepatocytes; Bc: blurred cell contours.

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图 2 慢性氨氮胁迫对幼鲫肾组织显微结构的影响

Te：肾小管；Gl：肾小球。

Figure 2. Effect of chronic ammonia nitrogen stress on microstructure of kidney tissue of juvenile C. auratus

Te: renal tubules; Gl: glomerulus.

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图 3 慢性氨氮胁迫对幼鲫肝、肾ALT的影响

不同小写字母表示同一胁迫时间不同处理组间差异显著（P＜0.05）。图4、5、6同。

Figure 3. Effect of chronic ammonia nitrogen stress on ALT in livers and kidneys of juvenile C. auratus

Data with different lowercase letters indicate significant difference among treatment groups in same duration under stress at P＜0.05. Same for Figs.4, 5 and 6.

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图 4 慢性氨氮胁迫对幼鲫肝、肾AST的影响

Figure 4. Effect of chronic ammonia nitrogen stress on AST in livers and kidneys of juvenile C. auratus

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图 5 慢性氨氮胁迫对幼鲫肝、肾CAT的影响

Figure 5. Effect of chronic ammonia nitrogen stress on CAT in livers and kidneys of juvenile C. auratus

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图 6 慢性氨氮胁迫对幼鲫肝、肾SOD的影响

Figure 6. Effect of chronic ammonia nitrogen stress on SOD in livers and kidneys of juvenile C. auratus

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表 1 氨氮对鲫幼鱼急性毒性试验结果

Table 1. Acute toxicity test of ammonia nitrogen on mortality of juvenile C. auratus

胁迫时间 Stress time/h	回归方程 Regressive equation	相关系数R²	半致死浓度 LC₅₀/(mg·L⁻¹)	95%置信区间 95% confidence interval/(mg·L⁻¹)	安全浓度SC/ (mg·L⁻¹)
24	P=5.7327X－15.0845	0.9939	427.86	393.45～465.28	42.8
48	P=7.7711X－19.9359	0.9985	367.62	356.67～378.91	36.8
72	P=8.6415X－21.7653	0.9879	330.14	271.14～401.98	33.0
96	P=5.5711X-13.7124	0.9980	289.29	264.08～316.90	28.9
LC₅₀采用概率单位法计算；P为死亡率；X为NH₄⁺质量浓度对数。 LC₅₀ was calculated by probabilistic unit method; P is mortality probability unit; X is log of NH₄⁺ concentration.

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