Experimental adaptation of human echovirus 11 to ultraviolet radiation leads to resistance to disinfection and ribavirin

Figures

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  Figure 1.. Experimental set-up. Echovirus 11 was propagated infecting Buffalo green monkey kidney (BGMK) cells in monolayers. The viruses were adapted to cell culture for three passages in the presence of ribavirin and then serially passaged after exposure to UVC or after an equivalent dilution between cell culture infections. The multiplicity of infection (MOI) in the experimental evolution passages was 0.0001.

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  Figure 2.. UVC inactivation of the wild-type (WT Rib+) and all the biological replicates of the population exposed to UVC (E01 to E04) after twenty passages of experimental adaptation. At minimum of three replicates were conducted for each population at initial concentrations of approximately 107 MPNCU/ml.

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  Figure 3.. Virus log inactivation obtained for the WT Rib+, NE01 and E01 after exposure to different UVC fluences. Asterisks indicate statistically significant differences compared to the wild-type (WT) Rib+. Different data points represent replicate experiments.

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  Figure 4.. Viral titre of wild-type (WT), WT Rib+ and evolved populations (E01 and NE01) obtained in the stationary growth phase (48–72 hours) in the presence of ribavirin. Data are presented as the ratio of the titre in the presence versus absence of the drug. Asterisks indicate a statistically significant difference with respect to the WT.

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  Figure 5.. Time-serial effective population sizes (Ne) and mutation rate per site per generation of NE01 (A) and E01 (B) populations. The orange lines indicate the median of 1000 Ne estimates between consecutive sampling time points with the given passages. The grey line shows the mutation rate calculated for each population. The dashed line indicates an increase in the UVC fluence.