To compare the optimum recoveries of an inoculation of <i>Salmonella enterica</i> serotype Typhimurium, fluorescent microspheres (1.0 μm diameter, carboxylate-modified, crimson FluoSpheres®, Molecular Probes, Eugene, OR), or a combination of both from stainless steel, three recovery methods, including a standard rinse, a one-ply composite tissue (Kimwipe®) or a sonicating brush were used. Findings were used to assess the effectiveness of fluorescent microspheres as surrogates for <i>S.</i> Typhimurium. For each method, ten coupons (304 grade, 2.5 x 8 cm) were inoculated with either 100 μl of a <i>S.</i> Typhimurium culture, or a solution of fluorescent microspheres, or both, at approximate concentrations of 10⁶. After drying for one hour, coupons were sampled using either a rinse of 100 ml of phosphate buffered saline solution (PBS) for one min, a Kimwipe® tissue method, or submerged in PBS and subjected to a sonicating brush for one min. After treatments, PBS solutions were analyzed using duplicate plate counting (<i>Salmonella</i>) or hemacytometry (microspheres). For microspheres and <i>Salmonella</i>, recovery by sonicating brush > rinse > Kimwipe® method. Additionally, the retention of microspheres on the steel ranged from 16 to 25% (mean from five coupons each recovery method). Microspheres yielded a significantly higher recovery rate (11 – 60%) than <i>Salmonella</i> (~1%) for each recovery method, therefore the microspheres used in this study, are not appropriate surrogates for <i>S.</i> Typhimurium for future recovery studies on stainless steel. However, due to their low standard deviations for their mean percent recovery, they hold the opportunity to provide better accuracy and reproducibility. / Master of Science

The ability of Listeria monocytogenes, to attach to various food contact surfaces such as stainless steel, polypropylene, and rubber compounds is well documented. The retention of these or other pathogenic bacteria on food contact surfaces increases the risk of transmission to food products. The objective of this study was to compare several methods for quantitative recovery of Listeria monocytogenes from stainless steel surfaces. A cocktail of four serotypes of Listeria monocytogenes (Scott A (4b)), 1/2b, 3b, and 4b) were mixed in equivalent concentrations and inoculated onto type 304 stainless steel coupons in a 2cm x 2cm area. After a one hour exposure, coupons were sampled by one of the following methods: 1) swabbing using a pre-moistened Dacron swab, 2) rinsing with phosphate buffered saline, 3) direct contact onto a Tryptic Soy Agar containing 0.6% yeast extract (TSA+YE) plate surface for 10 seconds, 4) sonication in an ultrasonic water bath (40 kHz), 5) contact with the bristles of a sonicating brush head for 1 min, and 6) indirect contact (2-4 mm) with the bristles of a sonicating brush head for 1 min. Coupon rinses were plated onto TSA containing 0.6% yeast extract and incubated for 24 hours at 35°C. The three sonication methods yielded higher recovery than the other three methods (p < 0.05). Brushing the coupons with the sonicating brush head yielded a recovery level of 58% and indirect exposure to the sonicating brush head permitted a recovery level of 65% from the initial microbial load. The lowest cell recovery (~20%) was observed with the swab and direct agar contact methods. / Master of Science