This study was conducted to detect the presence of Listeria monocytogenes (L. monocytogenes)
and screen for its antibiotic susceptibility characteristic from wildlife and water samples at
Kubah National Park, Sarawak, Malaysia. Samples collected were incubated and streaked on
selective medium PALCAM agar to confirm the presence of Listeria spp. before they were
further tested using molecular analysis. Specific Polymerase Chain Reaction (PCR) assay were
performed to target specific virulence gene, haemolysin gene, hlyA to further distinguish the
presence of this pathogenic bacteria in the samples. Overall, out of the 30 samples tested, 10
samples were confirmed as to contain L. monocytogenes strains and selected to subsequent
antibiotic susceptibility test. Susceptibility patterns to 10 antibiotics were investigated
among the L. monocytogenes strains. All strains were uniformly resistant to tetracycline and
erythromycin. On the other hand, all strains were sensitive to gentamycin and tobramycin. The
multiple antibiotic resistance shown by the strains in this study indicate the potential health
hazard associated with the possible transmission between wildlife and water to its surrounding
environment especially visitors and workers of Kubah National Park, Sarawak, Malaysia.