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Rapid Quantification of Planktonic Ciliates: Comparison of Improved Live Counting with Other Methods

机译:快速定量浮游纤毛虫:与其他方法改进的实时计数的比较

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摘要

The following efficient and quantitatively valid method to filter concentrate and count live planktonic ciliates was developed and compared with other treatments: unconcentrated (raw) samples and centrifuged samples were counted live, and the effects of five different fixatives (HgCl2, Lugol's iodine, formaldehyde, glutaraldehyde, and Champy-DaFano) on the counts were monitored. Samples originated from a eutrophic mountain lake (Lake Aydat, near Clermont-Ferrand, France). Overall, live filtered counts were similar to counts of raw samples, but they were significantly higher (2 to 2.3 fold, P < 0.05) by analysis of variance than counts from centrifuged samples. Nevertheless, some taxa, i.e., Halteria and Loxodes spp., were sensitive to filtration. The live filtered counts were also comparable to counts of raw HgCl2-fixed and settled samples. HgCl2 and Lugol fixation consistently gave the highest total counts, while significantly lower counts were always obtained with Champy-DaFano-fixed samples. Losses due to fixation were insignificant for raw samples but were substantial and statistically significant in concentrated samples (15% after filtration and 71% after centrifugation, compared with counts from the corresponding live samples). Live counting of passively filter-concentrated ciliates has many advantages over other methods. It is two to four times quicker and more efficient. Ciliates are recognized with certainty, more species are identified, and enumeration of dead organisms (e.g., tintinnid loricas) is avoided. It should be recommended as a quantitatively valid alternative to classical methods for assessing planktonic ciliate populations.
机译:开发了以下有效且在定量上有效的方法来过滤浓缩液并计数活的浮游纤毛虫,并将其与其他处理方法进行比较:对非浓缩(原始)样品和离心样品进行实时计数,以及五种不同的固定剂(HgCl2,Lugol的碘,甲醛,监测戊二醛和Champy-DaFano的计数。样品来自富营养化的高山湖泊(法国克莱蒙费朗附近的艾达特湖)。总体而言,实时过滤后的计数与原始样品的计数相似,但通过方差分析得出的结果明显高于离心样品的计数(2-2.3倍,P <0.05)。但是,某些分类单元,即Halteria和Loxodes spp。,对过滤很敏感。实时过滤后的计数也可与原始HgCl2固定和沉降样品的计数进行比较。 HgCl2和Lugol固定始终提供最高的总计数,而使用Champy-DaFano固定的样品始终获得低得多的计数。固定样品引起的固着损失微不足道,但在浓缩样品中则是显着且具有统计学意义的(与相应的活样品相比,过滤后为15%,离心后为71%)。相对于其他方法,对过滤器集中的纤毛进行实时计数具有许多优势。速度和效率提高了2到4倍。可以肯定地发现纤毛虫,鉴定出更多的物种,并且避免了死生物的计数(例如丁丁萝莉卡斯)。应该推荐它作为定量评估浮游纤毛虫种群的经典方法的定量替代方法。

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