Survival of Local Isolates of Enterobacter sakazakii (Cronobacter spp.) in Skim Milk during Spray Drying, Storage and Reconstitution

Abstract

Enterobacter sakazakii (Cronobacter spp.) is an emerging opportunistic pathogen associated with necrotizing enterocolitis, septicemia, bacteremia, and meningitis in neonates, particularly those who are premature or immunocompromised. Contaminated powder infant formula (PIF) has been linked to such outbreaks. Cronobacter spp. is relatively more resistant to osmotic stress and desiccation than other Enterobacteriaceae. This pathogen is known to survive at least for two years in PIF having low water activity (aw). The objective of this study was to determine the effects of spray drying and storage humidity (50%, 70%, and 90%) on the survival of a heat resistant local isolate Cronobacter spp. in skim milk and their viability upon reconstitution at various water temperatures. Survival of Cronobacter spp. during spray drying was determined by comparing the number of bacteria before and after spray drying. Viability of Cronobacter spp. in spray dried milk was observed for 3 month, during which Cronobacter spp. were periodically enumerated on TSA-YE agar. At the same intervals, spray dried milk containing Cronobacter spp. was reconstituted with water at room temperature (27 °C) and 50 °C. The number of Cronobacter spp. after reconstitution was enumerated and compared to the number of surviving Cronobacter spp. in milk prior to drying. In addition water activity (aw) and moisture content of the dried skim milk were also determined. Spray drying process resulted in 4.19 log CFU/g reduction of Cronobacter sp.YRc3a. Decrease in aw of skim milk after drying and during storage resulted in an increased survival of Cronobacter sp.YRc3a upon reconstitution at 50 °C. The logarithmic reduction of Cronobacter sp.YRc3a during reconstitution before drying were significantly higher (0.64 log) than after drying (0.35 log). Increase in survival upon reconstitution was also observed during storage at various humidity, ranging from 0.161 to 0.203 log. During 3 month storage period, Cronobacter sp. YRc3a survived better in dried skim milk at 70% humidity with only 0.21 log reduction in viable counts. Storage of dried skim milk at 50% and 90% humidity accelerated the rate of Cronobacter sp.YRc3a, resulting in decline of viable count for approximately 3 log cycles.