OBJECTIVE: To evaluate the cocktail effect of ACM, MTZ, and NMS during zebrafish\'s initial development.
METHODS: Zebrafish embryos 6-8 h post-fertilization (hpf) were exposed to different concentrations of ACM, MTZ, and NMS, separately, to obtain the 50% lethal concentrations (LC50). Next, the embryos were exposed to distinct concentrations of the cocktail (LC50/2, LC50/5, LC50/10, and LC50/20) in a semi-static system. Samples were analyzed 0, 24, 48, and 96 h after exposure, and the drugs\' concentrations in E3 medium were assessed by high-performance liquid chromatography. For embryotoxicity evaluation, the mortality, hatching, and heart rates; total length; and pericardial and yolk sac areas were determined. In addition, body malformations, edemas, presence of pigmentation, and histopathological assessments were also recorded.
RESULTS: The LC50 values obtained for MTZ, ACM, and NMS were 4.69 mgmL-1, 799.98 μgmL-1, and 0.92 μgmL-1, respectively. No difference was observed between the drugs\' nominal and observed concentrations at each time point. The cocktail significantly induced mortality and decreased hatching in the LC50/10, LC50/5, and LC50/2 groups. Additionally, body malformations, pigmentation loss, and yolk sac and pericardial edemas were observed in the cocktail groups. The cocktail groups\' larvae had decreased total length and slower heart rates compared to the controls (p < 0.05). The histopathological assessment showed that yolk sac edema promoted severe histological changes in the esophageal-intestine junction and intestine in larvae treated with cocktails. Moreover, PAS-positive structures decreased in the esophageal-intestine junction, intestine, and liver in larvae exposed to pharmaceutical cocktails.
CONCLUSIONS: This study\'s findings suggest the cocktail of ACM, MTZ, and NMS may be hazardous to aquatic organisms in case of environmental contamination.
目的:为了评估ACM的鸡尾酒效应,MTZ,和NMS在斑马鱼的初始发育过程中。
方法:斑马鱼胚胎受精后6-8小时(hpf)暴露于不同浓度的ACM,MTZ,NMS,分开,以获得50%致死浓度(LC50)。接下来,胚胎在半静态系统中暴露于不同浓度的混合物(LC50/2,LC50/5,LC50/10和LC50/20).在暴露后0、24、48和96小时分析样品,通过高效液相色谱法评估E3培养基中的药物浓度。对于胚胎毒性评估,死亡率,孵化,和心率;总长度;和心包和卵黄囊面积测定。此外,身体畸形,edemas,色素沉着的存在,并记录组织病理学评估.
结果:为MTZ获得的LC50值,ACM,和NMS分别为4.69mgmL-1、799.98μgmL-1和0.92μgmL-1。在每个时间点的药物标称浓度和观察浓度之间没有观察到差异。在LC50/10,LC50/5和LC50/2组中,鸡尾酒显着诱导死亡率和孵化减少。此外,身体畸形,色素沉着丧失,在鸡尾酒组中观察到卵黄囊和心包水肿。与对照组相比,鸡尾酒组幼虫的总长度减少,心率减慢(p<0.05)。组织病理学评估表明,卵黄囊水肿促进了用鸡尾酒处理的幼虫的食管-肠交界处和肠的严重组织学变化。此外,食管-肠连接处的PAS阳性结构减少,肠,和接触药物鸡尾酒的幼虫的肝脏。
结论:这项研究的发现表明ACM的混合物,MTZ,在环境污染的情况下,NMS可能对水生生物有害。