Anti-Methicillin Resistant Staphylococcus aureus (MRSA) Activity of Pseudoalteromonas flavipulchra Isolated from Marine Waters of Batangas, Philippines

Arizaldo Enriquez Castro, Cristina C. Salibay
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Abstract

The ocean boasts untapped novel producers of antibiotic substances in the form of unicellular organisms. One of the newest bioproducers of pharmacologically-significant substances studied for its potential against clinically-significant pathogens is the genus Pseudoalteromonas, a gammaproteobacterial group. This study is a preliminary report detailing the isolation of a Pseudoalteromonas flavipulchra species from Philippine marine waters. The isolate coded as PAM-003 was identified as 100 % similar to P. flavipulchra strain NCIMB2033 through 16s rRNA gene amplification and sequencing. PAM-003 was allowed to produce bioactives for 12 days. Afterwards, non-polar products were isolated from the base medium through membrane filtration, organic solvent extraction and rotary evaporation. The crude solution of bioactives injected in sterile discs was used for disc-diffusion assay against Methicillin Resistant Staphylococcus aureus (MRSA). Results indicate that PAM-003 demonstrated visually-appreciable zones of inhibition with a mean value of 8 mm. To further describe the antibacterial activity of the isolate, minimum inhibitory concentration (MIC) of the bacterial extract was determined through broth microdilution technique. Results indicate that PAM-003 demonstrated a MIC of 1000 µg/mL against MRSA. Additional investigation on the bioactivity of Philippine isolates from the genus Pseudoalteromonas isolated from highly diverse regions of the country is a considerable initiative for increasing the pipeline of new molecular entities for drug discovery.
菲律宾八打雁海域黄假互变单胞菌抗甲氧西林耐药金黄色葡萄球菌(MRSA)活性研究
海洋以未开发的单细胞生物形式的新型抗生素物质生产者而自豪。一个最新的具有药理意义的物质的生物生产者研究其对临床重要病原体的潜力是假互变单胞菌属,一个γ变形菌群。本研究是在菲律宾海水中分离到一株黄斑假互变单胞菌的初步报道。通过16s rRNA基因扩增和测序,鉴定分离物PAM-003与P. flavipulchra菌株NCIMB2033同源性100%。PAM-003被允许产生生物活性12天。然后,通过膜过滤、有机溶剂萃取和旋转蒸发从基础介质中分离出非极性产物。采用无菌圆盘注射生物活性物质粗液,进行了耐甲氧西林金黄色葡萄球菌(MRSA)的圆盘扩散试验。结果表明,PAM-003具有明显的视觉抑制区,平均为8 mm。为了进一步描述菌株的抑菌活性,采用肉汤微量稀释法测定了细菌提取物的最低抑菌浓度(MIC)。结果表明,PAM-003对MRSA的MIC为1000µg/mL。从菲律宾高度多样化的地区分离的假互交单胞菌属分离物的生物活性的进一步调查是增加药物发现新分子实体管道的重要举措。
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