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Trial Number 3

Client Number 153

Project Number 12

Trial Purpose:

To evaluate the efficacy of hydrogen peroxide cleaners with inactivating MS2 on a hard surface without agitation.

Date Run:

02/23/2024

Experiment Procedure:

Pour Plate Method - MS2 Bacteriophage

Six hours prior to the run, E.coli 15597 was subcultured into three milliliters of tryptic soy broth (TSB) screw-cap tubes and incubated at 37°C (98.6°F). 27 screw-cap tubes filled with 10ml of 0.5X tryptic soy agar (TSA) were autoclaved. The biosafety cabinet (BSC) was sprayed with 70% v/v isopropyl alcohol using a paper towel before spraying any items going into the BSC. Once autoclaving was complete, the TSA tubes were placed into a 45°C (113°F) D.I. water bath inside the biosafety cabinet (BSC). The four glass Petri dishes were marked using a black sharpie to designate the positive (P+), negative (N-), Test 1 (T1), and Test 2 (T2). Ten microliters of the organism were pipetted onto the P+, T1, and T2 stainless steel coupons and air-dried for 15 minutes. A motorized pipette with 10ml tips was used to pipet 15 ml of Dey-Engley (D/E) neutralizing broth into four separate 50ml conical tubes labeled P+, N-, T1, and T2. Once the MS2 bacteriophage dried on the coupons, the P+ coupon was placed into the conical tube. The N-, T1, and T2 were pipetted with 1000μl of the cleaning solution onto each coupon for 30 seconds before immediately placing them in the conical tube with an autoclaved forceps. The conical tubes were then placed on the shaker for 10 minutes. During this time, using the 1000ml pipette, 900ml of 1x phosphate-buffered saline (PBS) was pipetted into nine autoclaved dilution tubes, and serial dilutions were made for P+, T1, and T2 up to 10-4 using 100μl of the shaken D/E broth. Once the six-hour sub-time was complete, the E. coli 15597 subculture was removed from the incubator for use. For each variable (N-, P+, T1, and T2), 100μl of the stock and serial dilutions of MS2 bacteriophage, and 100μl of the E.coli 15597 subculture were combined into an empty dilution tube. A screwcap tube of 0.5X TSA was removed from the water bath, wiped with a paper towel to remove moisture, and poured into the dilution tube. The mixture was immediately poured into a sterile polystyrene petri dish; swirled to cover the entire plate surface, and then air-dried before covering. Dried Petri dishes were placed into a clean labeled zip lock bag that was partially closed and incubated at 37°C overnight. Plates were counted the following day based on the clear lysis zones in the bacterial lawn of growth (1 plate forming unit) to calculate log reduction and percent removal. 

Trial Results:

Product Log Reduction Percent Reduction
Bona Power Plus  Antibacterial Surface Cleaner 7.3500 100.0000
Bona Power Plus Hard-Surface Floor Cleaner 6.9275 100.0000
Earth Essentials Multi-purpose Disinfectant Cleaner 7.1400 100.0000
Homesolv by Citra Solv Multi-purpose disinfectant cleaner 7.3167 100.0000
Honest Disinfecting Spray (Without chlorine Bleach) 7.1833 100.0000
Kaboom (with the power of oxi-clean) 6.0943 99.9412
Libman Multisurface Disinfecting Cleaner 7.3133 100.0000
Lysol with Hydrogen Peroxide Multi-Purpose Cleaner  6.5900 100.0000
Oxivir 1 Disinfectant Cleaner 7.3675 89.7584
PERdiem® General Purpose Cleaner with Hydrogen Peroxide (Concentrate)  7.4233 100.0000
PERdiem General Purpose Cleaner with Hydrogen Peroxide (1:64 Dilution - Spray Bottle) 0.3626 55.6196
Puracy Disinfecting Surface Cleaner 6.8900 100.0000

Success Rating:

Results successful using TACT (time, agitation, concentration, and temperature, as well as rinsing and drying) and/or other cleaning chemistries examined.

Conclusion:

Bona Power Plus Antibacterial Surface cleaner, Bona power plus hard surface floor cleaner, earth essentials, Homesolv, Honest, Libman, Lysol, concentrated PERdiem, and Puracy had 100% MS2 reduction. Diluted PERdiem had a 55.6196% reduction, Oxivir had a 89.7574% reduction, and Kaboom had a 99.9412% reduction. 

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