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Title: TEMPERATURE AND NUTRIENT LIMITATION EFFECTS ON CAMPYLOBACTER JEJUNI ATTACHMENT AND SURVIVAL IN MULTISPECIES BIOFILMS ON STAINLESS STEEL

Author
item Sanders, Sheriase
item JOSEPH, FRANK - UGA-FOOD SCIENCE
item Arnold, Judy

Submitted to: Journal of Food Protection
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/31/2007
Publication Date: 2/1/2008
Citation: Sanders, S.Q., Joseph, F.F., Arnold, J.W. 2008. Temperature and nutrient limitation effects on campylobacter jejuni attachment and survival in multispecies biofilms on stainless steel. Journal of Food Protection. Vol 71:271-278.

Interpretive Summary: Campylobacter jejuni is a bacterium that causes food-borne illness in humans and is commonly found in the intestinal tract of chickens. In this study, attachment and survival of C. jejuni on stainless steel were assessed at different temperatures and with nutrient limitation. Bacterial isolates collected from a saline rinse of processed broiler chicken carcasses (WCR) were used to form communities of bacteria (biofilms). WCR biofilms were formed by incubation for 16 h at 13, 20, 37 and 42°C on stainless steel coupons in broth (TSB) for growth. The resulting biofilms were stained and visualized by using specialized microscopy. Biofilms formed at 13°C (47.6%) yielded the highest surface area coverage of cells with the lowest at 42°C (2.1%). C. jejuni was allowed to attach to 16-h biofilms at each of the previous temperatures and attached cells were counted. Attachment of C. jejuni did not significantly differ among the four temperatures. For nutrient limitation observations, WCR biofilms were produced in 1:10 and 1:50 diluted TSB at 20 and 37°C for 48 h. Surface area coverage was highest (approximately 2%) on coupons at 37°C for both TSB concentrations. Numbers of C. jejuni attached after 48 h to WCR biofilms at 20 and 37°C were determined. Attachment was different only for the 1:10 TSB (higher) at 20°C and 1:50 TSB at 37°C treatments. C. jejuni was cultured from coupons with biofilms incubated at 13 and 20°C, but not at 37 and 42°C. Also, C. jejuni was cultured from biofilms incubated at 20°C but not 37°C under nutrient limited conditions. These results show that the presence and survival of C. jejuni on stainless steel and in biofilms are affected by a combination of temperature and nutrient availability.

Technical Abstract: Campylobacter jejuni is a thermophilic, microaerophilic pathogen that is commonly found in the intestinal tract of chickens. In this study, attachment and survival of C. jejuni in biofilms on stainless steel were assessed at different temperatures and with nutrient limitation. Bacterial isolates collected from a saline rinse of processed broiler chicken carcasses (WCR) were used to form biofilms. WCR biofilms were formed by incubation for 16 h at 13, 20, 37 and 42°C on stainless steel coupons in tryptic soy broth (TSB). The resulting biofilms were stained with Hoechst (33258) and visualized by using epifluorescence microscopy. Biofilms formed at 13°C (47.6%) yielded the highest surface area coverage of cells with the lowest at 42°C (2.1%). C. jejuni, transformed to produce green fluorescent protein (gfp), was allowed to attach to 16-h biofilms at each of the previous temperatures and attached cells were enumerated. Attachment of C. jejuni did not significantly differ (P=0.05) among the four temperatures. For nutrient limitation observations, WCR biofilms were produced in 1:10 and 1:50 diluted TSB at 20 and 37°C for 48h. Surface area coverage was highest (approximately 2%) on coupons at 37°C for both TSB concentrations. Numbers of C. jejuni attached after 48h to WCR biofilms at 20 and 37°C, were determined. Attachment was different only for the 1:10 TSB (higher, P=0.05) at 20°C and 1:50 TSB at 37°C treatments. C. jejuni was cultured from coupons with biofilms incubated at 13 and 20°C, but not at 37 and 42°C. Also, C. jejuni was cultured from biofilms incubated at 20°C but not 37°C under nutrient limited conditions. These results show that the presence and survival of C. jejuni on stainless steel and in biofilms is affected by a combination of temperature and nutrient availability.