-
In this study, the disinfectant remained liquid without crystallization or precipitation in cold storage times of 72 h.
When evaluating the bactericidal effect of –18 °C disinfectant, the mean available chlorine concentration prepared in the laboratory tests was 2,919 mg/L (2,872–2,943 mg/L) while in field tests was 2,795 mg/L (2,765–2,819 mg/L). The available chlorine concentrations of –40 °C disinfectant before the 3 sets of laboratory experiments were 4,892 mg/L, 4,875 mg/L, and 4,839 mg/L, while those for field tests were 5,283 mg/L, 5,141 mg/L, and 5,247 mg/L.
-
For the two cryogenic disinfectants, two types of neutralizers could effectively neutralize the effect of the disinfectant remaining on the surface of bacteria in diluent on Staphylococcus aureus, and neither the neutralizer nor the neutralizing product had any adverse effect on Staphylococcus aureus or the medium (Table 1).
Group –18 °C Cryogenic disinfectant –40 °C Cryogenic disinfectant Average colony forming units of
each group (CFU/tablet)Error rate between groups (%) Average colony forming units of
each group (CFU/tablet)Error rate between groups (%) 1 407 4.2 542 4.23 2 413 592 3 450 602 4 0 0 Note: Group 1: soak the carrier tablet in 5.0 mL neutralizing agent for 10 minutes; Group 2: soak the carrier tablet in 5.0 mL of neutralization products for 10 minutes; Group 3: soak the carrier tablet in 5.0 mL tryptone saline solution (TPS) dilution for 10 minutes; Group 4: 1 mL of TPS diluent, 1 mL of neutralizing agent and the same culture medium as that of the first three groups was added and placed into sterile plates.The neutralizer of –18 °C Cryogenic disinfectant was 0.03 mol/L PBS containing 0.3% sodium thiosulfate, 0.1% lecithin and 1% tween-80 while 0.03 mol/L PBS containing 0.5% sodium thiosulfate, 0.5% lecithin and 2% tween-80 was for –40 °C Cryogenic disinfectant. Table 1. Neutralizer identification test on Staphylococcus aureus.
Based on neutralizer identification test, we carried out bactericidal experiments with the disinfectants. The average killing log values of the 2 disinfectants against Staphylococcus aureus and Escherichia coli were >3.00 at –18 °C and –40 °C, demonstrating the qualified effects of the disinfectants (Table 2).
Bacteria Available chlorine (mg/L) Logarithm of the mean colony number of the positive control group Average killing log of bacteria at different times of disinfection 2.5/5 min 5/10 min 7.5/15 min Staphylococcus aureus 2,919/4,869 6.18/6.07 5.81/3.91 5.88/4.12 6.18/4.10 Escherichia coli 2,919/4,869 6.36/6.36 5.23/4.78 5.23/5.14 6.18/5.39 Table 2. Killing effect of disinfectant of –18 °C/–40 °C disinfectant on Staphylococcus aureus and Escherichia coli.
-
There were 1 sample of Staphylococcus aureus and 1 sample of Escherichia coli on each side of each carton, and a total of 60 samples were collected at –18 °C or –40 °C. After the carton was sterilized by a high-speed micron spray sterilizer, the killing log values of Staphylococcus aureus and Escherichia coli on the carrier were all >3.00, demonstrating a qualified disinfection effect.
The killing log values of 30 Staphylococcus aureus tablets and 30 Escherichia coli tablets were all >3.00 after sterilizing the –18 °C or –40 °C storages with a sprayer (the dosage of disinfectant was 200–300 mL/m2), also meeting the requirements for qualified disinfectants (Tables 3–4).
Sample number Carrier position Colony forming units of test group (CFU/tablet) Killing log value Sample number Carrier position Colony forming units of test group (CFU/tablet) Killing log value 1 Top side of carton 1 (265, 0)/(10, 25) >3.00 16 Right side of carton 3 (10, 0)/(20, 5) >3.00 2 Bottom side of carton 1 (235, 15)/(615, 0) >3.00 17 Front side of carton 3 (5, 10)/(30, 0) >3.00 3 Left side of carton 1 (15, 0)/(10, 15) >3.00 18 Back side of carton 3 (0, 0)/(5, 0) >3.00 4 Right side of carton 1 (15, 0)/(10, 0) >3.00 19 Top side of carton 4 (0, 15)/(65, 0) >3.00 5 Front side of carton 1 (5, 10)/(50, 30) >3.00 20 Bottom side of carton 4 (425, 65)/(60, 950) >3.00 6 Back side of carton 1 (0, 5)/(0, 0) >3.00 21 Left side of carton 4 (0, 0)/(5, 0) >3.00 7 Top side of carton 2 (0, 15)/(480, 45) >3.00 22 Right side of carton 4 (10, 0)/(0, 0) >3.00 8 Bottom side of carton 2 (10, 50)/(515, 20) >3.00 23 Front side of carton 4 (5, 0)/(10, 0) >3.00 9 Left side of carton 2 (145, 0)/(5, 0) >3.00 24 Back side of carton 4 (10, 0)/(0, 0) >3.00 10 Right side of carton 2 (5, 0)/(5, 25) >3.00 25 Top side of carton 5 (0, 0)/(5, 5) >3.00 11 Front side of carton 2 (0, 10)/(5, 55) >3.00 26 Bottom side of carton 5 (15, 40)/(210, 5) >3.00 12 Back side of carton 2 (0, 20)/(5, 0) >3.00 27 Left side of carton 5 (50, 0)/(0, 5) >3.00 13 Top side of carton 3 (5, 0)/(10, 10) >3.00 28 Right side of carton 5 (5, 0)/(0, 15) >3.00 14 Bottom side of carton 3 (100, 5)/(0, 10) >3.00 29 Front side of carton 5 (0, 0)/(10, 0) >3.00 15 Left side of carton 3 (0, 0)/(10, 0) >3.00 30 Back side of carton 5 (230, 0)/(0, 0) >3.00 Note: Positive control bacteria count of Staphylococcus aureus and Escherichia coli: 4.26×106 CFU/tablet (9.0×105–6.6×106 CFU/tablet), 1.91×106 CFU/tablet (3.15×105–9.15×106 CFU/tablet). Table 3. The disinfection effect of –18 °C/–40 °C cryogenic disinfectant on Staphylococcus aureus/Escherichia coli on outer surface of the cartons.
Sample number Colony forming units of test group (CFU/tablet) Killing log value Sample number Colony forming units of test group (CFU/tablet) Killing log value 1 (0, 0)/(30, 5) >3.00 16 (5, 0)/(0, 5) >3.00 2 (0, 0)/(0, 0) >3.00 17 (0, 0)/(20, 10) >3.00 3 (0, 0)/(5, 0) >3.00 18 (0, 0)/(0, 0) >3.00 4 (0, 0)/(0, 65) >3.00 19 (0, 0)/(0, 0) >3.00 5 (0, 0)/(10, 5) >3.00 20 (0, 0)/(0, 70) >3.00 6 (0, 0)/(10, 0) >3.00 21 (0, 0)/(0, 65) >3.00 7 (0, 0)/(0, 0) >3.00 22 (0, 0)/(0, 0) >3.00 8 (0, 0)/(0, 0) >3.00 23 (0, 0)/(15, 135) >3.00 9 (0, 0)/(0, 0) >3.00 24 (0, 0)/(5, 115) >3.00 10 (0, 0)/(0, 0) >3.00 25 (0, 0)/(20, 10) >3.00 11 (0, 0)/(0, 15) >3.00 26 (0, 0)/(10, 100) >3.00 12 (0, 0)/(5, 0) >3.00 27 (0, 0)/(20, 0) >3.00 13 (0, 0)/(0, 0) >3.00 28 (0, 0)/(30, 0) >3.00 14 (0, 0)/(5, 0) >3.00 29 (0, 0)/(20, 340) >3.00 15 (0, 0)/(0, 0) >3.00 30 (0, 0)/(0, 170) >3.00 Note; Positive control bacteria count of Staphylococcus aureus and Escherichia coli: 4.26×106 CFU/tablet (9.0×105–6.6×106 CFU/tablet), 1.91×106 CFU/tablet (3.15×105–9.15×106 CFU/tablet). Table 4. The disinfection effect of –18 °C/–40 °C cryogenic disinfectant on Staphylococcus aureus/Escherichia coli in –18 °C/–40 °C storage.
-
With a combination of freezing tests, laboratory experiments and field tests, we obtained two disinfectants, the disinfectant for –18 °C contained 25% calcium chloride, 9.5% ethanol, and 3,000 mg/L sodium dichloroisocyanurate, while 30% calcium chloride, 9.5% ethanol, 9.9% ethylene glycol, 5,000 mg/L sodium dichloroisocyanurate, and 0.09% benzalkonium chloride were used in the –40 °C disinfectant.
HTML
Physical Property Tests
Laboratory Bactericidal Experiment
Field Test
Formulation of Disinfectant
Citation: |