Liaqat Iram, Mirza Safdar Ali, Iqbal Riffat, Ali Nazish Mazhar, Saleem Gulbeena, Majid Samia, Shahid Maryam
Department of Zoology, Govt. College University, Lahore, Pakistan.
Department of Botany, Govt. College University, Lahore, Pakistan.
Pak J Pharm Sci. 2018 Sep;31(5(Supplementary)):2047-2052.
Bacteria live either independently as planktonic cells or in organized surface associated colonies called as biofilms. Biofilms play an important role in increased pathogenesis of bacteria and it is assumed that motility is one of the contributing factors towards biofilm initiation. This study was planned to identify the role of flagella in biofilm formation by constructing flagellated (wild type) and physically disrupted variants (non-motile). Total 10 clinical bacterial strains were isolated and characterized. Morphological and biochemical study identified these strains as Enterobacter spp., Pseudomonas spp., Yersinia spp., Escherichia spp., Salmonella spp., Proteus spp., Staphylococcus spp., Streptococcus spp., Lactobacillus spp. and Bacillus spp. Among all strains, two strains including Yersinia spp and Bacillus spp. showed higher antibiotic resistance, hence studied at molecular and physiological level. Biofilm formation capacity of strains was analyzed using three methods including Congo red assay, Test tube assay and Liquid-interface coverslip assay. Afterwards, flagellar disintegration was induced by blending and centrifugation for 5, 10 and 15 minutes. 16S rRNA sequencing showed two strains as Bacillus cereus and Yersinia enterocolitica. Both strains produced significant biofilm by all three above mentioned methods. A motility test of these blended variants showed partial/diminished motility with increased blending time. The significant loss in biofilm formation after 15 minutes blending confirmed the important flagellar contribution to the initiation of biofilm formation. This biofilm defect observed in flagella paralysed/minus variants presumably may be due to defects in attachments to surface at early stages. This study indicated that flagellar motility is crucial initially for surface attachment and subsequently for biofilm formation.
细菌要么作为浮游细胞独立生存,要么形成称为生物膜的有组织的表面相关菌落。生物膜在细菌致病性增强中起重要作用,并且据推测运动性是生物膜形成的促成因素之一。本研究旨在通过构建有鞭毛的(野生型)和物理破坏的变体(不运动型)来确定鞭毛在生物膜形成中的作用。总共分离并鉴定了10株临床细菌菌株。形态学和生化研究将这些菌株鉴定为肠杆菌属、假单胞菌属、耶尔森菌属、埃希氏菌属、沙门氏菌属、变形杆菌属、葡萄球菌属、链球菌属、乳杆菌属和芽孢杆菌属。在所有菌株中,包括耶尔森菌属和芽孢杆菌属在内的两株菌株表现出较高的抗生素抗性,因此在分子和生理水平上进行了研究。使用刚果红试验、试管试验和液-界面盖玻片试验三种方法分析了菌株的生物膜形成能力。之后,通过搅拌和离心5、10和15分钟诱导鞭毛解体。16S rRNA测序显示两株菌株为蜡样芽孢杆菌和小肠结肠炎耶尔森菌。这两株菌株通过上述所有三种方法都产生了显著的生物膜。这些搅拌变体的运动性测试表明,随着搅拌时间的增加,运动性部分/减弱。搅拌15分钟后生物膜形成的显著损失证实了鞭毛对生物膜形成起始的重要贡献。在鞭毛麻痹/缺失变体中观察到的这种生物膜缺陷可能是由于早期表面附着缺陷所致。这项研究表明,鞭毛运动性最初对于表面附着至关重要,随后对于生物膜形成也至关重要。