An optical apparatus (Photonia^® manufactured by Biolight corporation) that emits light of ultra weak intensity was used, and its irradiance and optical spectrum were adjusted by passive optical components.

The measured irradiance at a position of 2 cm away from the center of the light emitting surface of the apparatus was about 5.2×10^-9 W/cm² and the dominant wavelength of the spectrum was 588 nm.

The PEDV strain DR-13 is a PEDV strain attenuated through serial passages resulting in a 51-nucleotide deletion in the ORF3 gene 7, and it is being used for a PED oral vaccine (DR13 10^6.5 TCID₅₀/mL) commercialised in 2003 by Green Cross Vet. Prod. (South Korea).

Virus titration was carried out using a 48-well microplate with Vero cells. Virus cultures were serially diluted 10-fold with the virus replication medium containing trypsin. Confluent Vero cells of the microplate were washed three times with PBS and inoculated at 0.1 mL per well into five wells. Following adsorption for 1 h at 37 ^oC, the inocula were removed, and the cells were washed three times with PBS. Subsequently, 0.1 mL of fresh virus replication medium containing trypsin was transferred into each well, and the cells were further incubated for 5 days at 37 8C. Fifty per cent tissue culture infective doses (TCID₅₀) were expressed as the reciprocal of the highest virus dilution showing cytopathic effect (CPE) 6 Antiviral effect of ultra weak light was expressed by inhibition titer, which was

calculated as follows: (log10(TCID₅₀/ml of control group) – log10(TCID₅₀/ml of experimental group)) 1

Set up an experimental group that exposed 10 ml of a pre-calculated (10^6.5 TCID₅₀/ml titer) PEDV to ultra weak light and that has not been exposed to light (control group) within the CO₂ 5% incubator. Light exposure times are 10, 24, 48, 72 and 96 hours, respectively.

After incubation of approximately 1,000 Vero cells in 25T flask in the CO2 5% incubator, the change in the number and pH of the cells is measured in the light treated experimental group and the non-lighted control group. Light exposure times are 10, 24, 48, 72 and 96 hours, respectively.

Incubate the 48 well plate Vero cells a day earlier and prepare an experimental group which exposed times (10, 24, 48, 72 and 96 hours) of ultra weak light and an unexposed control group.

The ultra weak light exposed experimental group after inoculation with the appropriate pre-calculate virus (titer; 10^5.3 TCID₅₀/ml) was compared with the non-exposed control group.

All experiments were repeated 3 times. Statistical comparisons were performed using ANOVA tests (p < 0.05) within the SPSS program (version 15.0.0) (SPSS Inc., USA). An asterisk (*) denotes statistical significance among the mean data.

We demonstrated changes in PEDV titers during exposure times for ultra weak light.

Following exposure to ultra weak light, the PEDV inhibition above the logarithmic reduction = log 0.1 (20.567%) from 10h to 72h was confirmed, but the statistical analysis was not significant (Figure 1). The effectiveness of disinfectant as a disinfectant announced a maximum reduced titer, in which the virus titer was reduced by at least 4 log10 25 was not met to be an effective dilution factor.

We examined how Vero cells changes following exposure to ultra weak light.

After exposure to ultra weak light, the pH level maintained lower than the control group for 10 to 96 h, which enabled Vero cells, increasing the number of cells by 1.2 to 1.5 times more than the control group. In particular, differences due to larger effects were identified during light exposure for over 72 hours, indicating that the statistical analysis was also significant (Figure 2). The ultra weak light activated the cell by decreasing the pH of the growth medium.

We found changes in PEDV in Vero cells exposed to ultra weak light.

After exposure to ultra weak light, the virus inhibition of log reduction=0.1 (20.567%) from 10h to 48h was confirmed, but statistical analysis showed no significant effects of virus reduction (Figure 3).