| dc.description.abstract | E.sakazakii is classified as a pathogenic bacterium that causes illness mostly to infants
and immunocompromised individuals. This bacterium belongs to the group of Gram negative
Enterobacteriaceae species, which was re-classified as Cronobacter spp. based on DNA
hybridization and phenotyping. E. sakazakii has been widely implicated in a number of foodborne
diseases, which are characterized by illnesses like meningitis and enteritis especially in
pre-termed neonates and or with low birth weight infants. The bacterium has been isolated from
various foods including fresh vegetables and fruits and also survives refrigeration temperatures.
This research aim was to isolate E.sakazakii from a number of fresh vegetables and fruits
samples available in local market surrounding Darmaga area, Bogor, Indonesia. The isolation
followed FDA (2002) method modified by Iversen and Forsythe (2004). Rapid biochemical kit,
API 20E, was used for biochemical characterization and identification. Molecular confirmation
consisted of isolation of DNA and PCR to amplify the gene encoding for 16S rRNA using two
different pair of primers, ESA-1/16SUNI-(R) and 16SUNI-L/Saka 2b. A total number of 19
samples tested positive to pre-enrichment and enrichment broth, in BPW (buffered peptone
water) and EE (Enterobacteriaceae enrichment) broth, respectively. Isolation in VRBG (Violet
Red Bile Glucose) agar yielded all positive results for all samples. However test on DFI
(Druggan–Forsythe–Iversen) agar showed only four isolates producing typical blue/green
colony, i.e. those isolated from cabbage, chaisin, apple “fuji” and apple “manalagi”. The
biochemical test kit API 20E (Analytical Profile Index; bioMe´rieux) showed that none of the
samples were positive for E.sakazakii. The PCR results showed that only one isolate i.e. from
cabbage, successfully produced an amplicon of the 16S rRNA. This study suggests that isolation
rate of E.sakazakii in vegetable is low at 5.2 %. | id |
