Effect of 17α-Ethinylestradiol on the Intestinal Calcium Carbonate Excretion Rate of Yellow Fin Sea Bream (Acanthopagrus latus)

N. Al Jandal
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Abstract

Marine fish maintain their hypotonic body fluids by drinking seawater, excreting excess salt, and retaining water. Calcium in seawater is precipitated as CaCO3 in marine fish gut to limit the Ca2+ availability for absorption from the ingested water to maintain Ca2+ homeostasis; this process is controlled by endocrine functions. This study was aimed to investigate the effect of the exposure to synthetic estrogen; ethinylestradiol (EE2), on the osmoregulation process (CaCO3 excretion rate from the intestine) and reproduction in the native fish species of Kuwait. The fish were exposed in a semi-static system to three treatments (control and 5 and 10 ng EE2/l of seawater) for two weeks, and then the terminal sampling was carried out. Blood samples were taken for vitellogenin (Vtg) analysis using Western blotting, dot blot, and immunohistochemistry (IHC). Calcium carbonate samples were analysed for the excretion rate and showed no significant difference in treatments indicating that no effect on osmoregulation was observed. Water samples were analyzed by the enzyme-linked immunosorbent assay (ELISA) for the concentration of EE2 in the exposure tanks, showing a 50% decrease in the nominal concentration after 24 h of exposure. Fish gonads and liver were dissected for histological examination, showing no obvious intersex in control and lowdose exposed group, whereas the high-dose exposed group showed signs of intersex. No Vtg was induced in the plasma and that could be due to many factors such as low concentrations received in the exposure tanks (2.3 and 4.9 ng/l), fish species, and exposure period. The hepatosomatic index showed no significant difference between treatments and the gonadosomatic index showed a significant difference between the control and high-dose group. Interesting results were obtained from IHC, where Vtg was localized in both treatments, although more Vtg was detected in higher-dose exposed fish for both tissues (liver and gonads).
17α-炔雌醇对黄鳍鲷肠道碳酸钙排泄率的影响
海鱼通过饮用海水、排泄多余的盐分和保留水分来维持低渗的体液。海水中的钙在海洋鱼类肠道中以CaCO3的形式沉淀,以限制钙离子从摄入水中的吸收,维持钙离子的稳态;这一过程受内分泌功能控制。本研究旨在探讨暴露于合成雌激素的影响;炔雌醇(EE2)对科威特本地鱼类渗透调节过程(肠道CaCO3排泄率)和繁殖的影响。在半静态系统中,将鱼暴露于三种处理(对照和5和10 ng EE2/l海水)中两周,然后进行终端采样。采集血样,采用Western blotting、dot blot和免疫组化(IHC)分析卵黄蛋白原(Vtg)。对碳酸钙样品的排泄率进行了分析,结果显示不同处理之间没有显著差异,表明对渗透调节没有影响。通过酶联免疫吸附试验(ELISA)分析水样暴露池中EE2的浓度,暴露24小时后标称浓度下降50%。解剖鱼的性腺和肝脏进行组织学检查,对照组和低剂量暴露组未见明显的双性,而高剂量暴露组有双性体征。血浆中未产生Vtg,这可能是由于许多因素造成的,例如暴露池中接受的低浓度(2.3和4.9 ng/l)、鱼类种类和暴露时间。肝体指数在治疗组间无显著差异,性腺指数在对照组和高剂量组间有显著差异。从免疫组化中获得了有趣的结果,尽管在高剂量暴露的鱼的两个组织(肝脏和性腺)中检测到更多的Vtg,但Vtg在两种处理中都是局部的。
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