Abstract Calcein is widely used to mark the new calcium carbonate structures in foraminifera, but it remains unclear whether calcein has effects on the biological features (i.e., abundance, size). Here the biological effects of calcein on benthic foraminifera were investigated using a two-factor crossed experiment in which foraminiferal communities (10,393 live specimens in total) were cultured in seawater spiked with calcein at four concentrations (control, 1, 5, and 10 mg/L) and for three incubation durations (2, 4, and 6 weeks). Each treatment was performed in triplicate, yielding 36 samples (4 concentrations × 3 durations × 3 replicates). Results showed that high calcein concentrations (10 mg/L) and long incubation times (6 weeks) significantly affected the abundance, and the Margalef and Shannon-Wiener diversity indices, of foraminiferal communities. Dominant species showed a species-specific response to calcein. Interestingly, different foraminiferal test types showed different responses to calcein: the proportion of miliolids significantly increased in all calcein treatments (1, 5, and 10 mg/L) relative to the control (without calcein), in contrast to the proportion of rotaliids. Thus, calcein might have a positive effect on foraminiferal communities by enhancing porcelaneous groups. Given that high calcein concentrations and long incubation times can have negative effects on foraminifera, we suggest a relatively safe manipulation: at the beginning of a culture experiment, calcein (≈5 mg/L) is added as an initial marker, then replaced with natural seawater halfway through the experiment. About 4 weeks before the end of the incubation, calcein can again be added to cultures.

中文翻译：

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不同浓度和培养时间下钙黄绿素掺入对底栖有孔虫群落的影响

摘要 钙黄绿素被广泛用于标记有孔虫新的碳酸钙结构，但钙黄绿素是否对生物特征（如丰度、大小）有影响尚不清楚。在这里，使用双因素交叉实验研究了钙黄绿素对底栖有孔虫的生物学影响，其中有孔虫群落（总共 10,393 个活标本）在掺有四种浓度（对照、1、5 和 10 毫克/毫克/ L) 和三个潜伏期（2、4 和 6 周）。每个处理一式三份进行，产生 36 个样品（4 个浓度 × 3 个持续时间 × 3 个重复）。结果表明，高钙黄绿素浓度 (10 mg/L) 和长孵育时间 (6 周) 显着影响了丰度，Margalef 和 Shannon-Wiener 多样性指数，有孔虫群落。优势物种对钙黄绿素表现出物种特异性反应。有趣的是，不同的有孔虫测试类型对钙黄绿素的反应不同：相对于对照（不含钙黄绿素），所有钙黄绿素处理（1、5 和 10 毫克/升）中粟粒类的比例显着增加，而轮状体的比例则相反。因此，钙黄绿素可能通过增强瓷群对有孔虫群落产生积极影响。鉴于高钙黄绿素浓度和长孵育时间会对有孔虫产生负面影响，我们建议进行相对安全的操作：在培养实验开始时，添加钙黄绿素 (≈5 mg/L) 作为初始标记，然后替换为天然实验中途的海水。在孵化结束前大约 4 周，