鲤鱼实验性铜中毒的病理学研究
摘要
本试验对鲤鱼进行了系统的铜急性中毒、铜亚急性中毒的病理学、血液学和组织蓄积性研究，从而探讨了铜用于水产养殖的安全性。以水中铜浓度分别为0.30mg/L、0.46mg/L、0.71mg/L、1.10mg/L、1.69mg/L、2.60mg/L、4.00mg/L对鲤鱼进行96h的急性毒性试验。采用寇氏法计算出24h、48h、96h的LC_(50)分别为1.67mg/L、1.25mg/L、0.77mg/L。安全浓度为0.077mg/L。中毒鱼体表和鳃的粘液增多，鳃上附着有淡蓝色的絮状物，出现神经症状，呼吸困难、严重者死亡。鳃丝增生呈棒状。肝细胞和肾小管上皮细胞颗粒变性、空泡变性、溶解性坏死。胰腺腺泡变性、坏死。肠粘膜上皮细胞变性。肠上皮细胞的线粒体肿胀、嵴断裂，整个线粒体呈空泡状。神经细胞内质网扩张、核糖体脱颗粒。以水中铜浓度分别为0.03mg/L、0.05mg/L、0.10mg/L、0.18mg/L、0.32mg/L对鲤鱼进行5周的亚急性毒性试验。各组发病率分别为5％、17.5％、25％、32.5％、52.5％，死亡率分别为0、2.5％、5％、10％、15％。中毒鱼的症状与急性试验鱼的相似，但是鳃上无淡蓝色絮状物，而且中毒鱼体色变黑，生长受到明显抑制。鳃小片上皮细胞增生、变性、坏死。心肌纤维空泡变性。肝细胞和肾小管上皮细胞空泡变性、溶解性坏死。胰腺凝固性坏死或溶解性坏死。肠粘膜层坏死。脑膜水肿、毛细血管充血、炎症细胞浸润。神经细胞变性、坏死。肝细胞核膜扩张、线粒体呈空泡状，溶酶体数量增多、体积增大。毛细胆管的微绒毛肿胀、断裂、脱落。肠上皮细胞的线粒体空泡化。肾小管上皮细胞核膜扩张，线粒体肿胀，嵴断裂。中毒鱼Hb、RBC减少，血浆SOD、CP、AKP活性降低，血浆GPT活性升高。微核试验结果是各试验组与对照组差异极显著或显著。这表明铜对鲤鱼有一定的遗传毒害性。用原子分光光度计检测中毒鱼肝、肾、鳃、肌肉中铜的含量，蓄积量的大小顺序是肝脏＞肾脏＞鳃＞肌肉。X射线显微分析结果表明：铜在肝细胞和肾细胞的溶酶体中分布最多，在细胞质中分布最少。
The systemic pathologies on acute, subacute copper poisoning in carp were first studied, which discussed the security of copper used in aquiculture. The 24h LC50,
48h LC50, 96h LC50 of acute poisoning were 1.67mg/L, 1.25mg/L, 0.77mg/L, respectively. Safe concentration was 0.077 mg/L. The mucus of body surface and gill increased in poisoned fish. There were some blue floccules in gills. Nervous sign, difficult breath and death were occurred in poisoned fish. The lesions of acute cases showed gill lamella hyperplasia like sticks; granular degeneration, vacuolar degeneration, necrosis in liver cells and renal tubular epithelial cells; pancreas degeneration, necrosis; intestinal mucous membrane epithelial cells degeneration. Mitochondria of liver cells swollen with disintegration and lysis of cristaes. Endoplasmic reticulum of neurocyte dilated with Shedding of ribosome particles. The subacute poisoning test was conducted for 5 weeks, with concentrations of 0.03mg/L, 0.05mg/L, 0.10mg/L, 0.18mg/L, 0.32mg/L.The morbidity and mortality were respectively 5%, 17.5%, 25%, 32.5%, 52.5% and 0, 2.5%, 5%, 10%, 15%. The clinic symptom of poisoned fish resembled acute poisoned fish',
but there was no blue floccule in gills. The colour of poisoned fish darkled, While the growth of poisoned fish was inhibited obviously. The lesions of subacute cases showed hyperplasia, degeneration, necrosis in the gill lamella epithelium; vacuolar degeneration in cardiac muscle fibre; vacuolar degeneration, lytic necrosis in the liver cells and renal tubular epithelial cells; coagulated necrosis or lytic necrosis in the glandular cells; intestinal epithelium cells necrosis; edema, hyperemia, hemorrhage in brain and degeneration, necrosis in neurocyte. The karyotheca dilated, mitochondria dissolved into vacuole, quantity of lysosome increased and volume of lysosome enlarged in liver cells. Microvilli of bile canaliculi sloughed and collapsed. Mitochondria dissolved into vacuole in intestinal mucous membrane epithelial cells. The karyotheca dilated and mitochondria swollen with disintegration and lysis of cristae in renal tubular epithelial cells. Haematologically, the red cell count and hemoglobin decreas
ed, serum SOD, CP, AKP
activity dropped; serum GPT activity arised. In microkernel experiment, the differences between control group and subacute poisoned groups were significant, which showed copper had hereditary toxicity to carp. Using atomic spectrophotometer examined copper contents of liver, kidney, muscle and gill in subacute poisoned fish. The sequence of accumulation was liver, kidney ,gill, muscle. Result of x-ray microanalysis showed that content of copper of microvillis was the most, while copper of cytosol was the least in liver cell and renal cell.
The systemic pathologies on acute, subacute copper poisoning in carp were first studied, which discussed the security of copper used in aquiculture. The 24h LC50,
48h LC50, 96h LC50 of acute poisoning were 1.67mg/L, 1.25mg/L, 0.77mg/L, respectively. Safe concentration was 0.077 mg/L. The mucus of body surface and gill increased in poisoned fish. There were some blue floccules in gills. Nervous sign, difficult breath and death were occurred in poisoned fish. The lesions of acute cases showed gill lamella hyperplasia like sticks; granular degeneration, vacuolar degeneration, necrosis in liver cells and renal tubular epithelial cells; pancreas degeneration, necrosis; intestinal mucous membrane epithelial cells degeneration. Mitochondria of liver cells swollen with disintegration and lysis of cristaes. Endoplasmic reticulum of neurocyte dilated with Shedding of ribosome particles. The subacute poisoning test was conducted for 5 weeks, with concentrations of 0.03mg/L, 0.05mg/L, 0.10mg/L, 0.18mg/L, 0.32mg/L.The morbidity and mortality were respectively 5%, 17.5%, 25%, 32.5%, 52.5% and 0, 2.5%, 5%, 10%, 15%. The clinic symptom of poisoned fish resembled acute poisoned fish',
but there was no blue floccule in gills. The colour of poisoned fish darkled, While the growth of poisoned fish was inhibited obviously. The lesions of subacute cases showed hyperplasia, degeneration, necrosis in the gill lamella epithelium; vacuolar degeneration in cardiac muscle fibre; vacuolar degeneration, lytic necrosis in the liver cells and renal tubular epithelial cells; coagulated necrosis or lytic necrosis in the glandular cells; intestinal epithelium cells necrosis; edema, hyperemia, hemorrhage in brain and degeneration, necrosis in neurocyte. The karyotheca dilated, mitochondria dissolved into vacuole, quantity of lysosome increased and volume of lysosome enlarged in liver cells. Microvilli of bile canaliculi sloughed and collapsed. Mitochondria dissolved into vacuole in intestinal mucous membrane epithelial cells. The karyotheca dilated and mitochondria swollen with disintegration and lysis of cristae in renal tubular epithelial cells. Haematologically, the red cell count and hemoglobin decreas
ed, serum SOD, CP, AKP
activity dropped; serum GPT activity arised. In microkernel experiment, the differences between control group and subacute poisoned groups were significant, which showed copper had hereditary toxicity to carp. Using atomic spectrophotometer examined copper contents of liver, kidney, muscle and gill in subacute poisoned fish. The sequence of accumulation was liver, kidney ,gill, muscle. Result of x-ray microanalysis showed that content of copper of microvillis was the most, while copper of cytosol was the least in liver cell and renal cell.
引文
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