• Azhar Rafique

• M. Luqman

• Zeeshan Nawaz

• Asma Ashraf

• Shabab Nasir

• Akhtar Rasool Asif

• M. Shahid Mahmood

• Rao Zahid Abbas

• Farhat Jabeen

• Tayyaba Sultana

• Salma Sultana

• Shabana Naz

• Farkhanda Asad

Abstract

Bacillus cereus is a gram positive, spore forming bacteria. It produces hemolytic and non-hemolytic enterotoxins, which lead to diarrhea, food poisoning and self-limiting gastrointestinal track (GIT) infections. The organism occurs worldwide, mostly the outbreaks due to B. cereus are linked to fried rice and milk products. The present study focused on detection of hemolytic (hblA, hblC) and non-hemolytic (nheA) toxin encoding gene subunits of B. cereus isolated form different milk samples. A total of 100 milk samples (refrigerated pasteurized, raw, heat treated and packed milk) were tested for the presence of B. cereus. Isolates were confirmed by biochemical testing. DNA was isolated through phenol chloroform method and confirmed by gel electrophoresis. For the detection of hemolytic and non-hemolytic subunits multiplex polymerase chain reaction (PCR) was performed. Out of 100 samples, 20% were found positive to B. cereus by biochemical testing. Among the 20% positive milk samples 50, 40 and 10% were heat treated refrigerated milk, raw milk and packaged milk, respectively. Results of multiplex PCR showed 100, 30 and 9% positive nheA, hblC and hblA genes subunits, respectively in positive isolates of B. cereus. Prevalence of B. cereus in milk could be a potential threat for milk consumers. Appropriate pasteurization and storage conditions must be followed to evade theB. cereus contamination.

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