PURPOSE The goal of this analysis was to determine the effectiveness of ozonated water as a hard surface sanitizer, knife dip sanitizer, and microbial reducer on meat cuts. These tests used 3M petrifilm to acquire data on microbial load reduction. This media is non-selective and allow for enumeration of all aerobic viable microorganisms present. Samples were analyzed following 10-15 second spray washing. The testing was performed from 8/21/01 through 3/13/02. MATERIALS Aqua Clean Mobile Sanitizer Sterile Latex Gloves 3M Aerobic Plate Count Petri Film Automatic Pipetter Permanent Marking Pen Micro 90 Cleaner Incubator, 34ºC Sterile 2ml Pipettes Sterile Transport Swabs with      10ml Letheen Broth       Hach Test Kit Steel Gloves Arm Guard Yellow Apron Processing Belts Cutting Knives Mezzanine Equipment (Final Grinder 5B, 6,        6B, 7, 8, 9 Hopper and Body) Contact Plates Boots Green Glove Air Knife Wizard Knife Hock Cutter Split Saw Brisket Saw 180ºF Water Tap Water Pig Foot Pig Ear Pig Hide Back Fat Jowls Belly Ham Shoulder Arm Pit METHODS All experiments were conducted in present working environments consisting of both kill floor and production areas. Samples were categorized as PPE equipment (steel glove, arm guard, yellow apron, green glove), cutting knives, organic material (foot, ear, hide, meat cuts), Mezzanine equipment, and cutting equipment (air knife, wizard knife, hock cutter, split saw, brisket saw). Samples were sprayed with ozonated water for approximately 10-15 seconds with an ozone concentration of 1.1 ppm to 1.4 ppm. Comparisons of microbial counts were performed on samples: Before and after ozonation Ozone vs. 180ºF water Ozone vs. 180ºF water and cold water Ozone alone. Swabbing was performed on all sample materials and transported to the microbiology laboratory. All samples were aseptically transferred to APC petrifilm and incubated for 48 hours. Counts were determined using a 10-1 dilution factor. Microbial counts 100 and greater were considered unacceptable. RESULTS PPE Equipment All PPE equipment that was tested showed a significant microbial reduction compared to before and after ozonation. A comparison of ozone vs. hot water on steel gloves showed that ozone performed better than 180ºF water (Tables 1, 3, 5) Cutting Knives All cutting knives showed a significant microbial reduction compared to before and after ozonation. For ozone vs. 180ºF water, both tests showed acceptable microbial reductions (Tables 2, 5, 8, 9). Organic material The contamination of pig ear, feet, and hide is extremely high, comparison of microbial counts before and after ozonation show a considerable reduction with some reduction to acceptable numbers. Various meat cuts taken from whole carcass showed significant acceptable microbial reduction (Tables 2, 3, 5, 6, 7,). Mezzanine Equipment Ozonated water sprayed on hoppers and grinders in the mezzanine area showed similar results of microbial counts compared to standard methods of sanitation (Table 4). Cutting Equipment - For comparison of ozone vs. 180ºF water, ozone performed as good as or better in reducing microbial load than 180ºF water. Microbial reduction was not observed for hock cutters for 180ºF water but significant reductions were observed for ozone (Tables 10, 11, 12). DISCUSSION All areas, equipment, and samples tested for this evaluation were chosen based on the criteria of heavy microbial load. An overall trend of reduction was observed. The aqua clean mobile sanitizer did have a significant effect on reducing the microorganism load of all areas tested and sterilization of some areas was achieved. From these results ozone can be used as a substitution of 180ºF water. Further experimentation will continue and the utilization of a permanent commercial ozone system enabling higher concentrations of ozone up to 3ppm will be used. Prediction of greater microbial reductions from higher ozone concentrations will be expected.   Table 1 - Testing of PPE w/ Ozone  (08/21/2001) Samples  Control  Test  Steel Glove  TNTC  130 Arm Guard  TNTC  230 Apron  TNTC  <10 Blue Arm Guard  TNTC  70 Belt 1   <10 Belt 2   <10 Belt 3   <10 Grease Fat on rail TNTC  <10   Table 2 - Ozone Test  (08/31/2001) Sample  Biotrace  Control  180 Water  Ozone Knife  500000  TNTC  4   Knife  25034  TNTC  0   Knife  44506  TNTC    0 Knife  36101  TNTC    3 Knife  4307  TNTC    *2 Steel Glove    TNTC  TNTC   Steel Glove    TNTC    2100 Pig foot    TNTC    **TNTC *Swabs taken 10 minute after spray with ozonated water ** too numerous to count but significantly less than control     Table 3 - Testing of PPE with Ozone (08/31/2001) Samples Control Test Steel Glove TNTC 290 Steel Glove TNTC 50 Yellow Apron TNTC 0 Yellow Apron TNTC 60 Arm Guard TNTC 10 Green Glove TNTC 100 Pigs Foot TNTC 10 Pigs Ear TNTC 1060 Hand   0     Date 8/15 8/17 8/20 8/28 8/31 9/5 9/7 9/11 9/14 9/19 31 Hopper 5 <10 <10 <10 <10 <10 290 <10 50 570 10 32 Grinder 6 400 TNTC <10 20 290 <10 TNTC <10 40 <10 33 Hopper 7 <10 <10 <10 <10 <10 <10 <10 <10 <10 <10 34 Grinder 8 <10 <10 <10 250 TNTC <10 TNTC <10 <10 <10 35 Link blend belt <10 <10 <10 <10 <10 TNTC TNTC <10 360 <10     = Ozone    = Sanitizer

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