Abstract
ObjectiveThe presence of Salmonella enterica serovar Typhimurium becomes a concern in relation to the safety of drinking water and ice. We detected and enumerated the bacteria from ice and beverages collected from several areas in Jakarta. Most Probable Number (MPN) and multiplex PCR method were used. Three sets of primers were used rfbJ, fliC, and fljB. Two Multiplex PCR’s were performed, the first is to detect the presence of Salmonella and the second is to confirm the positive isolate of S. enterica serovar Typhimurium.ResultsA total of 50 beverages and 50 ices were collected MPN result ranged from < 3 to > 11,000 MPN/ml. The highest MPN value > 11,000 MPN/ml. The first Multiplex PCR result from beverages, 58% positively contained Salmonella spp. with amplification of fliB gene and no amplification of rfbJ and fliC genes. For ice samples, 2% positively contained Salmonella spp. with rfbJ gene amplification, 62% fliB gene and no amplification of fliC gene. The second Multiplex PCR results from beverages identified 21 positive isolates of S. enterica serovar Typhimurium. In which, 17 isolates contained fljB gene and 4 isolates contained both fljB and rfbJ genes. From ice, 17 isolates having both rfbJ and fljB genes.
Highlights
ResultsA total of 50 beverages and 50 ices were collected Most Probable Number (MPN) result ranged from < 3 to > 11,000 MPN/ml
The first Multiplex Polymerase Chain Reaction (PCR) result from beverages, 58% positively contained Salmonella spp. with amplification of fliB gene and no amplification of rfbJ and fliC genes
Isolation of S. enterica Serovar Typhimurium and second multiplex PCR To confirm the presence of Salmonella, positive RV broth culture were diluted from 100 to 10−4 in 0.85% NaCl spread onto Xylose Lysine Desoxycholate agar (XLD) (OXOID, Hamsphire, England) and incubated at 37 °C for 24 h
Summary
Ice samples from North Jakarta, East Jakarta, Central Jakarta, and South Jakarta showed the same number of Salmonella spp, which was 2400 MPN/ml. Ice samples from West Jakarta showed a different number of Salmonella spp., which was > 11,000 MPN/ml (I.WJ. isolate). The highest MPN value > 11,000 MPN/ ml was found in sample from East Jakarta (B.EJ.). One sample from East Jakarta (I.EJ.2) was found to carry two amplicons from fljB and rfbJ genes. A total of 568 suspected colonies of S. enterica serovar Typhimurium were obtained from 31 ice positive samples. One isolate (2.5%) was obtained from I.EJ. sample and 16 positive isolates (66.67%) were obtained from I.EJ. sample All of these isolates were obtained from ice samples of East Jakarta (Table 2). The highest percentage of resistance was shown by streptomycin (10 μg) and erythromycin (15 μg)
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