Evaluation of High-Pressure Processing in Inactivation of the Hepatitis E Virus

Figures

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  FIGURE 1. Serial dilutions of HEV-3 strain 47832c were inoculated in triplicates onto A549/D3 cells. The HEV genome copy numbers in the supernatant was quantified at 14 d.p.i. using ddRT-PCR, and the mean copy number and the standard deviation (error bars) of three replicates each are shown. Correlation coefficient (R2) between the inoculated HEV-3c strain 47832c (genome copy number) and the harvested extracellular HEV (genome copy number) 14 d.p.i. in A549/D3 cells is demonstrated.

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  FIGURE 2. The effect of HPP treatment on HEV-3c strain 47832c in cell culture media. The samples containing 2 × 106 RNA copies were treated at 400 and 600 MPa for 1 and 5 min at ambient temperature in triplicates and were inoculated onto A549/D3 cells. The HEV genome copy numbers in the supernatant were quantified at 14 d.p.i. using ddRT-PCR. The effect is shown in comparison to the untreated viral stock. The standard deviation (error bars) of three replicates is shown.

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  FIGURE 3. The effect of HPP treatment on HEV-3c strain 47832c in ready-to-eat pork pâté. The samples containing 4 × 107 RNA copies of HEV were treated at 400 and 600 MPa for 1 and 5 min at ambient temperature in triplicates and were inoculated onto A549/D3 cells. The HEV genome copy numbers in the supernatant were quantified at 14 d.p.i. using ddRT-PCR. The effect is shown in comparison to the untreated but inoculated samples. Error bars represent standard deviation.

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  FIGURE 4. Amino acid sequence alignment of the N domain of the capsid protein (ORF2). The sequences for input, the untreated, treatment at 600 MPa for 1 min hold time and 600 MPa for 5 min hold time were aligned using Mega6 software and translated into amino acid sequences using BioEdit software.