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. 2023 Jan 30;14:1094877. doi: 10.3389/fmicb.2023.1094877

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Copyright © 2023 Nisar, Ross, Brown, Bentham, Best and Whiley.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Characterization of thermally generated (50°C/10 min) VBNC L. pneumophila. (A) Cytogram representing thiazole orange-stained alive (blue), thiazole orange-stained injured (green) and propidium iodide-stained dead (orange) cells generated by 50°C heat shock for 10 min. (B) These three fractions were cultured on BCYE agar, and their ATP contents measured using a luciferase-based ATP estimation assay. (C): FISH of A. polyphaga with Alexa Fluor 647 labeled EUK1209 (red) and Legionella with Alexa Fluor 488 labeled LEG705 (green) probes. (Ci) uninfected A. polyphaga. (Cii,iii) These micrographs show intracellular proliferation of VBNC Legionella within A. polyphaga after 7 and 10 days of infection, respectively.