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从污水中分离出的噬菌体可消除并防止生物膜的形成。

Bacteriophage Isolated from Sewage Eliminates and Prevents the Establishment of Biofilm.

作者信息

Ribeiro Karla Veloso Gonçalves, Ribeiro Cleberson, Dias Roberto Sousa, Cardoso Silvia Almeida, de Paula Sergio Oliveira, Zanuncio Jose Cola, de Oliveira Leandro Licursi

机构信息

Núcleo de Microscopia e Microanálise, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brasil.

Departamento de Biologia Geral, Universidade Federal de Viçosa, 36570-900, Viçosa, Minas Gerais, Brasil.

出版信息

Adv Pharm Bull. 2018 Mar;8(1):85-95. doi: 10.15171/apb.2018.011. Epub 2018 Mar 18.

Abstract

Biofilm growth exerts a negative impact on industry and health, necessitating the development of strategies to control. The objective of this work was study the lytic activity of the phage isolated from the sewage network in the formation and degradation of Escherichia coli biofilms. E. coli cultures were incubated in 96-well polystyrene microplates under controlled conditions to evaluate the biofilm formation. The E. coli cultures and established biofilms were treated with the suspensions of the vB_EcoM-UFV017 (EcoM017) bacteriophage obtained from sewage for 24 hours. The E. coli bacterial density was measured using absorbance at 600 nm and the biofilms were measured by crystal violet staining. Polystyrene coupons were used as support for Scanning Electron Microscopy and Confocal Microscopy to evaluate biofilm formation. The E. coli strains formed biofilms in polystyrene microplates after 48 hours' incubation. The highest EcoM017 phage titer, in the prevention and degradation experiments, reduced the bacterial growth and the quantity of biofilm formed by E. coli in 90.0% and 87.5%, respectively. The minimum dose capable of reducing the biofilms of this bacterium was 10 PFU/mL after 24 hours. The preformed E. coli biofilm mass was reduced 79% post exposure to the phage in the degradation assay. Microscopic analysis confirmed the results obtained in the plates assays. The EcoM017 phage prevented biofilm formation and degraded the E. coli-established ones. The EcoM017 phage isolated from sewage can reduce bacterial attachment and lyse the E. coli associated biofilm cells, offering biotechnological potential applicability for this phage.

摘要

生物膜生长对工业和健康产生负面影响,因此有必要制定控制策略。这项工作的目的是研究从污水管网中分离出的噬菌体对大肠杆菌生物膜形成和降解的裂解活性。将大肠杆菌培养物在96孔聚苯乙烯微孔板中于受控条件下孵育,以评估生物膜的形成。用从污水中获得的vB_EcoM-UFV017(EcoM017)噬菌体悬浮液处理大肠杆菌培养物和已形成的生物膜24小时。使用600nm处的吸光度测量大肠杆菌的细菌密度,并用结晶紫染色测量生物膜。使用聚苯乙烯试片作为扫描电子显微镜和共聚焦显微镜的支撑物,以评估生物膜的形成。孵育48小时后,大肠杆菌菌株在聚苯乙烯微孔板中形成生物膜。在预防和降解实验中,EcoM017噬菌体的最高滴度分别使大肠杆菌的细菌生长和生物膜形成量减少了90.0%和87.5%。24小时后,能够减少这种细菌生物膜的最小剂量为10 PFU/mL。在降解试验中,预先形成的大肠杆菌生物膜质量在暴露于噬菌体后减少了79%。显微镜分析证实了平板试验中获得的结果。EcoM017噬菌体可防止生物膜形成并降解已形成的大肠杆菌生物膜。从污水中分离出的EcoM017噬菌体可减少细菌附着并裂解与大肠杆菌相关的生物膜细胞,为该噬菌体提供了生物技术潜在应用价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ac8/5896399/4f4edba4e572/apb-8-85-g001.jpg

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