3. Finally, surface test was tried once more using 10 mg and 20 mg of Stalosan powder with four different contact times (5, 10, and 60 minutes and 6 hours). The difference in this experiment was that, after 90 minutes of incubation with different dilutions of the samples, the inoculated cells were washed twice with PBS (phosphate buffered saline) followed by incubation at 37C. This approach was successful; no CPE was visible in any of the sample. This experiment was done in triplicate and the results are shown in Tables 1-3. Table 1. Effect of Stalosan F powder on PEDV (Experiment 1) Sanitizer A B Time of Contact 5 min 10 min 60 min 6 hour 5 min 10 min 60 min 6 hour Virus Titer (log 10 TCID 50 ) in: Control Stalosan F 4.17 <1 4.50 <1 4.17 <1 4.83 <1 4.50 <1 4.50 <1 4.17 <1 4.83 <1 Percent virus reduction ≥ 99.93 ≥ 99. 96 ≥ 99.93 ≥ 99. 99 ≥ 99. 96 ≥ 99. 96 ≥ 99.93 ≥ 99. 99 A: 10 mg Stalosan F powder Control A: 10 mg beef extract powder B: 20 mg Stalosan F powder Control B: 20 mg beef extract powder TCID 50 : 50% tissue culture infective dose of virus Table 2. Effect of Stalosan F powder on PEDV (Experiment 2) Sanitizer Time of Contact Virus Titer (log 10 Percent virus reduction TCID 50 ) in: Control Stalosan F A 5 min 10 min 60 min 6 hour B 5 min 10 min 60 min 6 hour 4.83 <1 4.50 <1 4.50 <1 4.17 <1 5.17 <1 4.50 <1 4.17 <1 4.50 <1 ≥ 99.99 ≥ 99. 96 ≥ 99. 96 ≥ 99. 93 ≥ 99. 99 ≥ 99. 96 ≥ 99.93 ≥ 99. 96 A: 10 mg Stalosan F powder Control A: 10 mg beef extract powder B: 20 mg Stalosan F powder Control B: 20 mg beef extract powder TCID 50 : 50% tissue culture infective dose of virus 3
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