Antimicrobial property and identification of bacteria in the mucus from the cloaca of green turtles(Chelonia mydas)

Abstract

This study was conducted in Chagar Hutang, Redang Island. Mucus samples secreted by the green turtle, Chelonia mydas during the oviposition process was collected using sterile universal bottles. The sediment samples surrounds the nesting area was also co!Jected. Mucus samples were brought back to the laboratory for screening of antimicrobial properties and also for identification of bacteria present. While the sediment sample was only analyse for the purpose of bacteria identification. Kirby-Bauer method was used to screen the antimicrobial properties in the mucus samples. No inhibition zone was observed meaning that there was no antimicrobial properties in the green turtle cloaca. On the other hand, the identification of bacteria present in the samples was first screened based on their morphological characteristics such as the shape, elevation, margin and also pigmentation. Once identified, the single pure colony was isolated for genotypic identification using the 16S rRNA (primers 63F and l 389R) cloning via Polymerase Chain Reaction. PCR products were then run in 1.5% of agarose gel for the formation of band at 1400Kb. Purified PCR products were sent for sequencing at the First BASE Laboratories Sdn Bhd. Sequences obtained were analyze using Chromas, BioEdit Sequence Alignment Editor and search for sequence similarity using NCBI BLAST. The two different bacteria identified from the mucus samples are Shewanella haliotis and Pseudoalteromonas spongiae. While the bacteria that identified from the sediment was Vibrio hepatarius.