The zebrafish (Danio rerio) is widely used as a powerful model organism in biomedical and drug screening research¹. With an exceptional capacity to assess multiple compounds per day, fish-based screenings have already been used to identify several drugs that reached preclinical and clinical trials². Zebrafish have also emerged as an important species for probing mechanisms of brain function and dysfunction. These studies include modeling multiple central nervous system (CNS) diseases³, such as depression, anxiety, psychosis, attention deficit hyperactivity disorder, autism, neurodegenerative disorders and addiction. Collectively, these applications highlight the potential of zebrafish models for neurobehavioral research and high-throughput genetic and small neuroactive molecule screening.

However, although data replicability and reproducibility are fundamentally important to consider in the field of zebrafish research⁴, multiple experimental factors remain poorly documented and evaluated. For example, many zebrafish studies do not report the water chemistry parameters of holding tanks, or the pH of drug treatments, in the methods sections. Water chemistry, especially ammonia levels, can strongly impact behavioral endpoints in zebrafish tests. In adult zebrafish, a 4-h exposure to 90 mg/L of NH4⁺ induces strong fear and decreases shoaling cohesion, and at 180 mg/L it lowers aggressiveness and social recognition, compared to unexposed controls⁵. Although these concentrations are higher than those generally found in fish maintenance tanks, these effects illustrate the overall (yet presently under-recognized) impact of environmental factors, such as water parameters and quality, on key zebrafish behaviors. The water salinity also strongly influences zebrafish behavior, as fish exposed to high-salinity home tank water (600 µS) stay closer to the bottom of their tanks than fish exposed to lower-salinity (50 µS) water⁶. Likewise, a change of a single pH unit (e.g., pH 7 to pH 8), within the pH ranges commonly used in zebrafish husbandry, is enough to alter larvae locomotor activity at a fixed drug concentration⁷.

斑马鱼（ Danio rerio ）作为强大的模式生物被广泛应用于生物医学和药物筛选研究¹ 。凭借每天评估多种化合物的卓越能力，基于鱼类的筛选已被用于鉴定多种已进入临床前和临床试验的药物² 。斑马鱼也已成为探索大脑功能和功能障碍机制的重要物种。这些研究包括对多种中枢神经系统 (CNS) 疾病进行建模³ ，例如抑郁症、焦虑症、精神病、注意力缺陷多动障碍、自闭症、神经退行性疾病和成瘾。总的来说，这些应用凸显了斑马鱼模型在神经行为研究以及高通量遗传和小神经活性分子筛选方面的潜力。

然而，尽管数据的可复制性和再现性在斑马鱼研究领域中至关重要⁴ ，但多个实验因素仍然缺乏记录和评估。例如，许多斑马鱼研究并未在方法部分报告储水池的水化学参数或药物处理的 pH 值。水化学，尤其是氨水平，可以强烈影响斑马鱼测试中的行为终点。在成年斑马鱼中，与未暴露的对照组相比，暴露于 90 mg/L NH4 ⁺ 4 小时会引起强烈的恐惧并降低浅滩凝聚力，而 180 mg/L 的 NH4 + 会降低攻击性和社会认可度⁵ 。尽管这些浓度高于鱼类维持池中通常存在的浓度，但这些影响说明了环境因素（例如水参数和质量）对斑马鱼关键行为的总体（但目前尚未得到充分认识）影响。水的盐度也强烈影响斑马鱼的行为，因为暴露于高盐度家庭水箱水 (600 µS) 的鱼比暴露于低盐度 (50 µS) 水的鱼更靠近水箱底部⁶ 。同样，在斑马鱼饲养中常用的 pH 范围内，单个 pH 单位（例如，pH 7 至 pH 8）的变化足以改变固定药物浓度下幼虫的运动活性⁷ 。