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In vivo assessment of pathogens toxicity on Daphnia magna using fluorescent dye staining

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Abstract

Daphnia has been widely used as an indicator species in aquatic biomonitoring for decades. Traditional toxicity assays based on lethality take a long time to assess, and the effect mode of contaminants is not clear. Because of the translucency of the Daphnia body and the application of fluorescent probes in cell staining, different intoxicated parts can be visualized. In this study, a double-staining method using two fluorescent dyes, Calcein AM (cell-permeant dye) and Propidium Iodide (cell-impermeant dye), was carried out on Daphnia magna exposed to six pathogens: Salmonella spp. (four strains) and Shigella spp. (two strains). The results showed that those bacteria caused different infections on daphnia depending on the age of this organism and bacterial concentrations. In detail, S. dublin and S. sonnei are the most harmful to Daphnia when they cause damage at smaller concentrations at the younger stage (3 weeks old). Interestingly, older Daphnia can give responses to nearly 10 CFU/ml to less than 100 CFU/ml of some bacteria strains. In another experiment, S. sonnei disturbed Daphnia after just 10 min of exposure, and Daphnia adapted to S. choleraesuis, S. typhi, and S. flexneri at the early stage (3 weeks old) after 1 h of exposure. Moreover, the damaged areas of the daphnia body were directly observed via a microscope, contributing to the understanding and the prediction of toxicity mechanisms.

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Funding

This research was supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (2020R1A6A1A06046235).

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Authors and Affiliations

  1. School of Chemical Engineering, Jeonbuk National University, 567 Baekje-daero, Deokjin-Gu, Jeonju-si, Jeollabuk-do, 54896, South Korea

    Vu Quynh Anh Le & Jiho Min

  2. Graduate School of Semiconductor and Chemical Engineering, Jeonbuk National University, 567 Baekje-daero, Deokjin-Gu, Jeonju-si, Jeollabuk-do, 54896, South Korea

    Wooil Choi, Taehwan Kim & Jiho Min

  3. Department of Food Science and Biotechnology, Shin Ansan University, 135 Sinansandaehak-Ro, Danwon-Gu, Ansan, 15435, South Korea

    Sung Min Woo

  4. School of Biological Sciences, Chungbuk National University, 1 Chungdae-Ro, Seowon-Gu, Cheongju, 28644, South Korea

    Yang-Hoon Kim

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  1. Vu Quynh Anh Le
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  2. Wooil Choi
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  3. Taehwan Kim
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  5. Yang-Hoon Kim
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Correspondence to Yang-Hoon Kim or Jiho Min.

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Le, V.Q.A., Choi, W., Kim, T. et al. In vivo assessment of pathogens toxicity on Daphnia magna using fluorescent dye staining. Ecotoxicology 29, 892–899 (2020). https://doi.org/10.1007/s10646-020-02257-6

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  • DOI: https://doi.org/10.1007/s10646-020-02257-6

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