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. 2020 Dec 15;203(4):1299–1308. doi: 10.1007/s00203-020-02125-1

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Survival of I. hospitalis for reproduction (culturable) and viable but non-culturable (VBNC) state. Black circles: viable and replication-competent, hence culturable. Gray circles: transition state between culturable, and viable but non-culturable (VBNC). Open circle: viable but non-culturable (VBNC). The experiment was conducted with n = 3 and refers to the results on lead acetate paper. Trendlines were fitted by hand. b Controls to discriminate between the culturable and viable but non-culturable state. Metabolic activity of three independently exposed (γ-radiation) I. hospitalis stationary phase cultures (IH1, IH2, and IH3) monitored on lead acetate paper. Exposed ½ SME + S⁰ medium, and exposed stationary phase cultures (1:10 diluted in fresh medium) were incubated at 90 °C for up to 6 days (columns 1, 3, and 4). Exposed IH1 stationary phase cultures were tenfold diluted in fresh medium before incubation (column 2). For higher sensitivity, 0.2 ml of the last positive IH1 bottle within a serial dilution were transferred into fresh medium, and incubated at 90 °C for up to 6 days (columns 5, 6). The same was conducted with IH2 (columns 7, 8), and IH3 (columns 9, 10). DLR laboratory control, BGS transport control. Serial dilutions with 1:10 dilution steps (10^–1–10^–8) are represented by the exponent (− 1 to − 8), and 2 and 6 refer to the days of incubation (Koschnitzki 2016)