1. Microbiology of Kitchen Sponges Jim Abshire, Maggie Crenshaw May 10, 2006 BSCI 223H

2. Introduction Kitchen sponges can be effective growth environments for bacteria –Continually wet environment provides ideal growing conditions –Repeated contact with food and [mess] adds nutrients and new bacteria Most common bacteria found on sponges –E. coli (from fecal matter, meat) –S. aureus (normal flora on some people’s skin) –Salmonella enterica

3. Introduction Sponges are sometimes microwaved in order to reduce bacterial concentrations –Heat (boils water) –Steam from evaporating water –Dries out sponge Must microwave until sponge is dry –Remaining bacteria can survive and multiply in greater numbers when sponge is wet

4. Research Questions How “dirty” are communal sponges used in residence halls? –What kind of bacteria are found in kitchen sponges used in Anne Arundel Hall? –How many? What effect does microwaving sponges have on bacterial counts? Hypothesis: Sponges will contain Salmonella, E. coli, and Staphylococcus

5. Materials 2 used, wet sponges TSA SS Agar (Salmonella-Shigella) Vogel-Johnson Agar (Staphylococcus) MacConkey’s Agar Sterile saline Sterile flasks, test tubes

6. Methods 1.Squeeze out liquid into sterile flask 2.Serial dilutions on TSA for viable cell counts a.0.9 mL saline into each tube b.0.1 mL culture into first tube c.Serial dilutions to 10 -6 d.Plated 0.1 mL from each tube on TSA 3.Spread 0.1 mL original culture onto 4 of each type of plate

7. Methods 4.Microwaved sponges until dry (3.5 min) 5.Re-wetted with 30 mL sterile saline 6.Squeezed out into new flasks 7.Repeated dilution and plating procedure 8.Incubated all cultures for 48 hours at 37˚C

8. Results Before Microwaving (TSA)After Microwaving (TSA)

9. Results Growth on all media Viable Plate Counts (CFU / mL) BeforeAfterReduction Sponge 17.7 x 10 7 2.3 x 10 3 99.995% Sponge 21.2 x 10 8 5.6 x 10 3 99.997%

10. Results SS agar before and after microwaving sponges

11. Results MacConkey’s agar before and after microwaving sponges

12. Results VJ agar before and after microwaving sponges

13. Discussion Presence of Salmonella, E. coli, Staphylococci, and other enterobacteria on sponges Microwaving dramatically decreased concentrations of enterobacteria –Concentration of staphylococci increased –Contact with hands shortly before incubation

14. E. coli Member of enterobacteriaceae –Gram negative, lactose fermenting, –Found commonly in intestinal tracts of animals As a pathogen: best known for causing intestinal infections Most infectious strain: E. coli O157:H7 –73,000 cases of infection each year in the US –61 deaths per year –Symptoms include bloody diarrhea, abdominal cramps, occasionally kidney failure –Most people recover without antibiotics within 5-10 days

15. Salmonella Members of enterobacteriaceae –99% of diseases caused by S. enterica –Live in intestinal tracts of animals –Gram-negative, oxidase-negative, motile Cause self-limiting gastroenteritis –Rehydrate patient (diarrhea) –Only treat with antibiotics if spread outside intestines Each year 40,000 cases reported –Only most severe cases reported –About 600 fatal cases per year

16. Staphylococcus Gram-positive cocci Normal flora in humans Some are pathogenic: S. aureus –Gastrointestinal illness: nausea, vomiting, cramps, diarrhea –Fast-acting toxin: illness within 30 min –Self-limiting, toxin not affected by antibiotics –Cause > 90% of reported food poisoning cases in US

17. Concluding Comments Protocol was sound: may have introduced staphylococcus by hand-squeezing sponges –May have been out-competed on sponges before microwaving Results supported hypothesis Further research would evaluate pathogenicity of bacteria found Kitchen sponges should be replaced frequently –Contain enteric (possibly pathogenic) bacteria –Microwaving is useful technique to reduce bacterial concentrations

18. References Josephson, K.L., Rubino, J.R., and Pepper, I.L, 1997. Characterization and quantification of bacterial pathogens and indicator organisms in household kitchens with and without the use of a disinfectant cleaner. Journal of Applied Microbiology. 83(6): 737-750. Sharp, K., and Walker, H., 2003. A microbiological survey of communal kitchens used by undergraduate students. International Journal of Consumer Studies. 27(1): 11-16. Ikawa, J., and Rossen, J., 1999. Reducing Bacteria in Household Sponges. Journal of Environmental Health. 62(1): 18-22. http://www.cdc.gov http://medinfo.ufl.edu/year2/mmid/bms5300/bugs/esccoli.html http://textbookofbacteriology.net/e.coli.html