Comparison of Cronobacter sakazakii from Agra region grown in biofilms, agar surface associated and planktonic mode by proteomic analysis

Context: Cronobacter sakazakii is an emerging food borne pathogen that causes severe meningitis, meningoencephalitis, sepsis, and necrotizing enterocolitis in neonates and infants, with a high fatality rate. Aims: The present paper is for the rapid detection of C. sakazakii from milk and milk products of Agra region via PCR method and comparison of C. sakazakii in biofilm, on agar surface and planktonic cells by proteomic analysis. Materials and Methods: In the present study, 55 samples of milk and milk products of the Agra region were analyzed. 200 isolates were obtained of which 11 were biochemically detected as C. sakazakii . The PCR targeting the ompA gene was used to amplify a 496 bp DNA segment unique to C. sakazakii , in order to confirm C. sakazakii isolates. The proteome was investigated to study the differential protein pattern expressed by biofilm, agar surface-associated and planktonic bacteria employing SDS-PAGE. Statistical Analysis: UN-SCAN-6.1 gel analysis software. Results: The primer pair ESSF and ESSR was successfully used to amplify a 469 bp DNA unique to C. sakazakii . Whole cell protein profiles of planktonic, biofilm and agar surface associated were characteristic. Conclusion: The cultural procedure for detection of C. sakazakii is laborious, taking up to 7 days for completion, whereas PCR combined with enrichment culturing can detect C. sakazakii in about 12 hours and thus has the potential to be used as a rapid tool for detecting its presence. Differential protein pattern of C. sakazakii cultivated in biofilm versus agar-surface-associated and planktonic cells were observed. Further understanding the role of specific proteins during the biofilm development should permit a better understanding of the mechanisms sustaining the proliferation and the resistance of bacteria on biotic surfaces.