Rehman Saba, Mujtaba Ghauri Shahbaz, Sabri Anjum Nasim
Department of Microbiology and Molecular Genetics, Quaid-e-Azam Campus, University of the Punjab, Lahore, Pakistan; Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan.
Department of Ear, Nose and Throat, Mayo Hospital, King Edward Medical University, Lahore, Pakistan.
Jundishapur J Microbiol. 2016 Jan 2;9(1):e29483. doi: 10.5812/jjm.29483. eCollection 2016 Jan.
Otitis media can lead to severe health consequences, and is the most common reason for antibiotic prescriptions and biofilm-mediated infections. However, the increased pattern of drug resistance in biofilm forming bacteria complicates the treatment of such infections.
This study was aimed to estimate the biofilm formation potential of the clinical isolates of otitis media, and to evaluate the efficacy of antibiotics and plant extracts as alternative therapeutic agents in biofilm eradication.
The ear swab samples collected from the otitis media patients visiting the Mayo Hospital in Lahore were processed to isolate the bacteria, which were characterized using morphological, biochemical, and molecular (16S rRNA ribotyping) techniques. Then, the minimum inhibitory concentrations (MICs) of the antibiotics and crude plant extracts were measured against the isolates. The cell surface hydrophobicity and biofilm formation potential were determined, both qualitatively and quantitatively, with and without antibiotics. Finally, the molecular characterization of the biofilm forming proteins was done by amplifying the ica operon.
Pseudomonas aeruginosa (KC417303-05), Staphylococcus hemolyticus (KC417306), and Staphylococcus hominis (KC417307) were isolated from the otitis media specimens. Among the crude plant extracts, Acacia arabica showed significant antibacterial characteristics (MIC up to 13 mg/ml), while these isolates exhibited sensitivity towards ciprofloxacin (MIC 0.2 µg/mL). All of the bacterial strains had hydrophobic cellular surfaces that helped in their adherence to abiotic surfaces, leading to strong biofilm formation potential (up to 7 days). Furthermore, the icaC gene encoding polysaccharide intercellular adhesion protein was amplified from S. hemolyticus.
The bacterial isolates exhibited strong biofilm formation potential, while the extracts of Acacia arabica significantly inhibited biofilm formation among the isolates and, therefore, could be executed in the development of cost-effective biofilm inhibitor medicines.
中耳炎可导致严重的健康后果，是抗生素处方和生物膜介导感染的最常见原因。然而，形成生物膜的细菌耐药性增加模式使此类感染的治疗复杂化。
本研究旨在评估中耳炎临床分离株形成生物膜的潜力，并评估抗生素和植物提取物作为生物膜根除替代治疗剂的疗效。
从拉合尔梅奥医院就诊的中耳炎患者采集耳拭子样本，进行细菌分离，采用形态学、生化和分子（16S rRNA基因分型）技术对其进行鉴定。然后，测定抗生素和植物粗提物对分离株的最低抑菌浓度（MIC）。定性和定量测定有无抗生素时细胞表面疏水性和生物膜形成潜力。最后，通过扩增ica操纵子对形成生物膜的蛋白质进行分子鉴定。
从中耳炎标本中分离出铜绿假单胞菌（KC417303 - 05）、溶血葡萄球菌（KC417306）和人葡萄球菌（KC417307）。在植物粗提物中，阿拉伯金合欢显示出显著的抗菌特性（MIC高达13 mg/ml），而这些分离株对环丙沙星敏感（MIC 0.2 μg/mL）。所有菌株的细胞表面具有疏水性，有助于它们粘附于非生物表面，导致强大的生物膜形成潜力（长达7天）。此外，从溶血葡萄球菌中扩增出编码多糖细胞间粘附蛋白的icaC基因。
细菌分离株表现出强大的生物膜形成潜力，而阿拉伯金合欢提取物显著抑制分离株中的生物膜形成，因此可用于开发具有成本效益的生物膜抑制剂药物。
