Antibacterial and Antibiofilm Potential of Sea Anemone (Stichodactyla haddoni) Isolated Vibrio (V. parahaemolyticus and V. alginolyticus), and Pseudoalteromonas (P. gelatinilytica and P. piscicida) Against Staphylococcus aureus and Pseudomonas aeruginosa

N. Fazeli, Akram Sadat Naeemi, S. Jalali, H. Zamani
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Abstract

Background: Staphylococcus aureus and Pseudomonas aeruginosa are important human bacterial pathogens, which are resistant to several antibiotics. One of the main causes of their resistance is the ability of biofilm formation. Objectives: The present study aimed to evaluate the antibacterial and antibiofilm activity of the extracts of Vibrio parahaemolyticus, V. alginolyticus, Pseudoalteromonas gelatinilytica, and Pseudoalteromonas piscicida isolated from sea anemone (Stichodactyla haddoni) against S. aureus and P. aeruginosa. Methods: Four isolated bacteria were identified using biochemical and molecular identification methods, and their extracts were obtained by mixing the cell-free supernatants from their old broth culture using ethyl acetate and methanol as the solvents. The agar well-diffusion and micro-dilution methods were also applied to determine the antibacterial activity, minimum bactericidal concentration (MBC), and minimum inhibitory concentration (MIC) of the extracts. The ability of the extracts to inhibit biofilm formation and disrupt the preformed biofilm of the pathogens was attained through crystal violet staining in 96-well microtiter plates. To determine the nature of the extracts, they were exposed to protease enzyme, and the antibiofilm activity was compared with the untreated extracts. Results: The extracts of the four isolated bacteria inhibited bacterial growth and biofilm formation and disrupted the preformed biofilm of S. aureus (MIC = BIC = 600 µg/mL) and P. aeruginosa (MIC = BIC = 300 µg/mL). In addition, the active compounds of the extracts with antibiofilm activities were mainly proteases. Conclusions: According to the results, V. parahaemolyticus, V. alginolyticus, P. gelatinilytica, and P. piscicida had antibacterial and antibiofilm potential against S. aureus and P. aeruginosa, and their extract could also be further analyzed as an alternative to antibiotics.
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背景:金黄色葡萄球菌和铜绿假单胞菌是人类重要的致病菌,对多种抗生素具有耐药性。它们产生耐药性的主要原因之一是形成生物膜的能力。目的:研究从海葵中分离得到的副溶血性弧菌、溶藻弧菌、明胶性假异单胞菌和鱼绿假异单胞菌提取物对金黄色葡萄球菌和铜绿假单胞菌的抑菌活性和抗膜活性。方法:采用生化和分子鉴定方法对4株分离细菌进行鉴定,以乙酸乙酯和甲醇为溶剂,将菌体老肉汤培养的无细胞上清液混合,得到菌体提取物。采用琼脂孔扩散法和微稀释法测定提取物的抑菌活性、最低杀菌浓度(MBC)和最低抑菌浓度(MIC)。通过96孔微滴板结晶紫染色,获得了提取物抑制生物膜形成和破坏病原体预先形成的生物膜的能力。为了确定提取物的性质,将其暴露于蛋白酶中,并与未处理的提取物进行抗生素活性比较。结果:4株分离菌提取物对金黄色葡萄球菌(MIC = BIC = 600µg/mL)和铜绿假单胞菌(MIC = BIC = 300µg/mL)的细菌生长和生物膜形成有抑制作用,对预形成的生物膜有破坏作用。此外,具有抗膜活性的活性化合物主要为蛋白酶。结论:副溶血性弧菌、溶藻弧菌、明胶溶菌和鱼酸弧菌对金黄色葡萄球菌和铜绿假单胞菌均具有抗菌和抗生物膜潜力,其提取物也可作为抗生素的替代品进行进一步研究。
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