Efficacy of gaseous chlorine dioxide in inactivating Bacillus cereus spores attached to and in a biofilm on stainless steel

Biofilm formation by Bacillus cereus 038-2 on stainless steel coupons, sporulation in the biofilm as affected by nutrient availability, temperature, and relative humidity, and the resistance of vegetative cells and spores in biofilm to sanitizers were investigated. Total counts in biofilm formed on coupons immersed in tryptic soy broth (TSB) at 12 and 22°C consisted of 99.94% of vegetative cells and 0.06% of spores. Coupons on which biofilm had formed were immersed in TSB or exposed to air with 100, 97, 93, or 85% relative humidity. Biofilm on coupons immersed in TSB at 12°C for an additional 6 days or 22°C for an additional 4 days contained 0.30 and 0.02% of spores, respectively, whereas biofilm exposed to air with 100 or 97% relative humidity at 22°C for 4 days contained 10 and 2.5% of spores, respectively. Sporulation did not occur in biofilm exposed to 93 or 85% relative humidity at 22°C. Treatment of biofilm on coupons that had been immersed in TSB at 22°C with chlorine (50 μg/ml), chlorine dioxide (50 μg/ml), and a peroxyacetic acid–based sanitizer (Tsunami 200, 40 μg/ml) for 5 min reduced total cell counts (vegetative cells plus spores) by 4.7, 3.0, and 3.8 log CFU per coupon, respectively; total cell counts in biofilm exposed to air with 100% relative humidity were reduced by 1.5, 2.4, and 1.1 log CFU per coupon, respectively, reflecting the presence of lower numbers of vegetative cells. Spores that survived treatment with chlorine dioxide had reduced resistance to heat. It is concluded that exposure of biofilm formed by B. cereus exposed to air at high relative humidity (≥97%) promotes the production of spores. Spores and, to a lesser extent, vegetative cells embedded in biofilm are protected against inactivation by sanitizers. Results provide new insights to developing strategies to achieve more effective sanitation programs to minimize risks associated with B. cereus in biofilm formed on food contact surfaces and on foods.

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Evaluation of Chlorine, Chlorine Dioxide, and a Peroxyacetic Acid–Based Sanitizer for Effectiveness in Killing Bacillus cereus and Bacillus thuringiensis Spores in Suspensions, on the Surface of Stainless Steel, and on Apples

Journal of Food Protection ◽

10.4315/0362-028x-69.8.1892 ◽

2006 ◽

Vol 69 (8) ◽

pp. 1892-1903 ◽

Cited By ~ 36

Author(s):

AUDREY C. KRESKE ◽

JEE-HOON RYU ◽

LARRY R. BEUCHAT

Keyword(s):

Stainless Steel ◽

Bacillus Thuringiensis ◽

Bacillus Cereus ◽

Chlorine Dioxide ◽

Food Processing ◽

Horse Serum ◽

Peroxyacetic Acid ◽

Food Processing Industry ◽

Bacillus Spores ◽

Food Contact Surface

Chlorine (10 to 200 μg/ml), chlorine dioxide (10 to 200 μg/ml), and a peroxyacetic acid–based sanitizer (40 and 80 μg/ml) were evaluated for effectiveness in killing spores of Bacillus cereus and Bacillus thuringiensis in suspensions and on the surface of stainless steel and apples. Water and 5% horse serum were used as carriers for spore inoculum applied to the surface of stainless steel coupons, and 5% horse serum was used as a carrier for inoculum applied to apples. Inocula were dried on stainless steel for 5 h and on apples for 22 to 24 h before treating with sanitizers. At the concentrations of sanitizers tested, sensitivities of planktonic B. cereus and B. thuringiensis spores were similar. A portion of the spores surviving treatment with chlorine and, more markedly, chlorine dioxide had decreased tolerance to heat. Planktonic spores of both species were more sensitive to sanitizers than were spores on the surface of stainless steel or apples. At the same concentrations, chlorine was more effective than chlorine dioxide in killing spores in suspension and on stainless steel. The lethality of chlorine dioxide was markedly reduced when inoculum on stainless steel coupons was suspended in 5% horse serum as a carrier rather than water. Chlorine and chlorine dioxide at concentrations of 10 to 100 μg/ml were equally effective in killing spores on apples. Significant reductions of ≥3.8 to 4.5 log CFU per apple were achieved by treatment with 100 μg/ml of either of the two sanitizers. The peroxyacetic acid sanitizer (40 and 80 μg/ml) was ineffective in killing Bacillus spores in the test systems investigated. Results provide information on the effectiveness of sanitizers commonly used in the food processing industry in killing Bacillus spores in suspension, on a food-contact surface, and on a ready-to-eat food.

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Lethality of Chlorine, Chlorine Dioxide, and a Commercial Produce Sanitizer to Bacillus cereus and Pseudomonas in a Liquid Detergent, on Stainless Steel, and in Biofilm

Journal of Food Protection ◽

10.4315/0362-028x-69.11.2621 ◽

2006 ◽

Vol 69 (11) ◽

pp. 2621-2634 ◽

Cited By ~ 29

Author(s):

AUDREY C. KRESKE ◽

JEE-HOON RYU ◽

CHARLES A. PETTIGREW ◽

LARRY R. BEUCHAT

Keyword(s):

Stainless Steel ◽

Bacillus Cereus ◽

Chlorine Dioxide ◽

Total Population ◽

Horse Serum ◽

Effective Strategies ◽

Food Contact Surfaces ◽

Dishwashing Detergent ◽

Contact Surfaces ◽

Spoilage Microorganisms

Many factors that are not fully understood may influence the effectiveness of sanitizer treatments for eliminating pathogens and spoilage microorganisms in food or detergent residues or in biofilms on food contact surfaces. This study was done to determine the sensitivities of Pseudomonas cells and Bacillus cereus cells and spores suspended in a liquid dishwashing detergent and inoculated onto the surface of stainless steel to treatment with chlorine, chlorine dioxide, and a commercial produce sanitizer (Fit). Cells and spores were incubated in a liquid dishwashing detergent for 16 to 18 h before treatment with sanitizers. At 50 μg/ml, chlorine dioxide killed a significantly higher number of Pseudomonas cells (3.82 log CFU/ml) than did chlorine (a reduction of 1.34 log CFU/ml). Stainless steel coupons were spot inoculated with Pseudomonas cells and B. cereus cells and spores, with water and 5% horse serum as carriers. Chlorine was more effective than chlorine dioxide in killing cells and spores of B. cereus suspended in horse serum. B. cereus biofilm on stainless steel coupons that were treated with chlorine dioxide or chlorine at 200 μg/ml had total population reductions (vegetative cells plus spores) of ≥4.42 log CFU per coupon; the number of spores was reduced by ≥3.80 log CFU per coupon. Fit (0.5%) was ineffective for killing spot-inoculated B. cereus and B. cereus in biofilm, but treatment with mixtures of Fit and chlorine dioxide caused greater reductions than did treatment with chlorine dioxide alone. In contrast, when chlorine was combined with Fit, the lethality of chlorine was completely lost. This study provides information on the survival and sanitizer sensitivity of Pseudomonas and B. cereus in a liquid dishwashing detergent, on the surface of stainless steel, and in a biofilm. This information will be useful for developing more effective strategies for cleaning and sanitizing contact surfaces in food preparation and processing environments.

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Disinfection of Fusarium-infected rice seeds by gaseous chlorine dioxide

Seed Science and Technology ◽

10.15258/sst.2014.42.3.02 ◽

2014 ◽

Vol 42 (3) ◽

pp. 322-331 ◽

Cited By ~ 2

Author(s):

Y.-A. Jeon ◽

S. Lee ◽

Y. Lee ◽

H.-S. Lee ◽

J.S. Sung ◽

...

Keyword(s):

Chlorine Dioxide ◽

Gaseous Chlorine Dioxide ◽

Rice Seeds

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Gaseous chlorine dioxide technology for improving microbial safety of spices

Innovative Food Science & Emerging Technologies ◽

10.1016/j.ifset.2021.102783 ◽

2021 ◽

pp. 102783

Author(s):

Xinyao Wei ◽

Tushar Verma ◽

Mary-Grace C. Danao ◽

Monica A. Ponder ◽

Jeyamkondan Subbiah

Keyword(s):

Chlorine Dioxide ◽

Microbial Safety ◽

Gaseous Chlorine Dioxide

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Evaluation of Gaseous Chlorine Dioxide as a Sanitizer for Killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and Yeasts and Molds on Fresh and Fresh-Cut Produce

Journal of Food Protection ◽

10.4315/0362-028x-68.6.1176 ◽

2005 ◽

Vol 68 (6) ◽

pp. 1176-1187 ◽

Cited By ~ 112

Author(s):

KAYE V. SY ◽

MELINDA B. MURRAY ◽

M. DAVID HARRISON ◽

LARRY R. BEUCHAT

Keyword(s):

Escherichia Coli ◽

Listeria Monocytogenes ◽

Chlorine Dioxide ◽

Foodborne Pathogens ◽

Escherichia Coli O157 ◽

Gaseous Chlorine Dioxide ◽

Sensory Qualities ◽

A Cell ◽

Fresh Cut ◽

Yeasts And Molds

Gaseous chlorine dioxide (ClO2) was evaluated for effectiveness in killing Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on fresh-cut lettuce, cabbage, and carrot and Salmonella, yeasts, and molds on apples, peaches, tomatoes, and onions. Inoculum (100 μl, ca. 6.8 log CFU) containing five serotypes of Salmonella enterica, five strains of E. coli O157:H7, or five strains of L. monocytogenes was deposited on the skin and cut surfaces of fresh-cut vegetables, dried for 30 min at 22°C, held for 20 h at 4°C, and then incubated for 30 min at 22°C before treatment. The skin surfaces of apples, peaches, tomatoes, and onions were inoculated with 100 μl of a cell suspension (ca. 8.0 log CFU) containing five serotypes of Salmonella, and inoculated produce was allowed to dry for 20 to 22 h at 22°C before treatment. Treatment with ClO2 at 4.1 mg/liter significantly (α = 0.05) reduced the population of foodborne pathogens on all produce. Reductions resulting from this treatment were 3.13 to 4.42 log CFU/g for fresh-cut cabbage, 5.15 to 5.88 log CFU/g for fresh-cut carrots, 1.53 to 1.58 log CFU/g for fresh-cut lettuce, 4.21 log CFU per apple, 4.33 log CFU per tomato, 1.94 log CFU per onion, and 3.23 log CFU per peach. The highest reductions in yeast and mold populations resulting from the same treatment were 1.68 log CFU per apple and 2.65 log CFU per peach. Populations of yeasts and molds on tomatoes and onions were not significantly reduced by treatment with 4.1 mg/liter ClO2. Substantial reductions in populations of pathogens on apples, tomatoes, and onions but not peaches or fresh-cut cabbage, carrot, and lettuce were achieved by treatment with gaseous ClO2 without markedly adverse effects on sensory qualities.

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