Survival of Campylobacter jejuni within mixed population biofilms of poultry isolates on stainless steel
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A method was developed to determine the attachment and survivability of Campylobacter jejuni in mixed biofilms on stainless steel. Confocal scanning laser microscopy (CSLM) and epifluorescence microscopy were used for visualization of C. jejuni transformed with a Pcgfp plasmid (C. jejuni1221gfp). The data in this study indicated that C. jejuni 1221gfp was able to form a biofilm on stainless steel when incubated with and without other bacteria at 37 °C up to 7 days. The numbers of C. jejuni1221gfp were enhanced on the stainless steel when incubated with mixed poultry isolates to form biofilms. C. jejuni1221gfp was visualized being attached to 16 hr biofilms formed on stainless steel at temperatures of 13, 20, 37 and 42 °C. The frequency of attachment at the four temperatures was statistically the same even though biofilm surface coverage was less at 37 and 42 °C when compared to 13 and 20 °C. Culturable C. jejuni1221gfp was recovered only from biofilms formed at 13 and 20 °C, however, according to epifluorescence microscopy cells were seen at 37 and 42 °C, attached to biofilm and stainless steel but they may be in the viable-but-nonculturable phase or nonviable. C. jejuni1221gfp was able to attach to stainless steel with preexisting biofilms and survive in nutrient limited environments. In diluted tryptic soy broth (1:10 and 1:50 TSB), biofilms of mixed culture were allowed to form and C. jejuni1221gfp was allowed to attach at 20 and 37 °C. Biofilm surface area coverage was low (approximately 2%) in both broths at both temperatures. Attached C. jejuni1221gfp differed in numbers between 1:10 TSB (20 °C) and 1:50 TSB (37 °C) with numbers from 1:10 TSB being the highest between the two samples. Counts of C. jejuni1221gfp were recovered only from 20 °C. Epifluorescent microscopy displayed C. jejuni1221gfp directly attaching to the coupon and not areas of biofilm in both broth and temperature treatment combinations in a 48 hr period.