REDPROM Centre (Recombinant DNA and Recombinant Protein Research Centre), Aydin Adnan Menderes University, Aydın, Turkey.
Aydın Vocational School of Health Services, Aydin Adnan Menderes University, Aydın, Turkey.
Arch Microbiol. 2022 May 30;204(6):353. doi: 10.1007/s00203-022-02962-2.
Phage DNA analysis gives opportunity to understand living ecosystem of the environment where the samples are taken. In the present study, we analyzed phage DNA obtained from wastewater sample of university hospital sewage. After filtration, long high-speed centrifugation was done to collect phages. DNA was extracted from pellet by phenol chloroform extraction and used for NGS sequencing. The host profile, taxonomic and functional analyses were performed using MG-RAST, and ResFinder program was used for resistance gene detection. High amounts of reads belong to bacteriophage groups (~ 95%) from our DNA sample were obtained and all bacteriophage reads were found belonging to Caudovirales order and Myoviridae (56%), Siphoviridae (43%), and Podoviridae (0.02%) families. The most common host genera were Escherichia (88.20%), Salmonella (5.49%) and Staphylococcus (5.19%). SEED subsystems hits were mostly structural parts and KEGG Orthology hits were nucleotide- and carbohydrate metabolism-related genes. No anti-microbial resistance genes were detected. Our bacteriophage DNA purification method is favorable for phage metagenomic studies. Dominance of coliphages may explain infrequent Podoviridae. Dominancy of structural genes and auxiliary genes is probably due to abundance of lytic phages in our sample. Absence of antibiotic resistance genes even in hospital environment phages indicates that phages are not important carrier of resistance genes.
噬菌体 DNA 分析使我们有机会了解样本采集环境中的微生物生态系统。在本研究中,我们分析了从大学医院污水废水样本中获得的噬菌体 DNA。经过过滤后,进行长时间高速离心以收集噬菌体。通过酚氯仿抽提从沉淀中提取 DNA,并用于 NGS 测序。使用 MG-RAST 进行宿主谱、分类和功能分析,使用 ResFinder 程序检测耐药基因。从我们的 DNA 样本中获得了大量属于噬菌体的序列(~95%),所有噬菌体序列都属于尾噬菌体目(Caudovirales order)和肌尾噬菌体科(Myoviridae)(56%)、长尾噬菌体科(Siphoviridae)(43%)和短尾噬菌体科(Podoviridae)(0.02%)。最常见的宿主属是大肠杆菌(Escherichia)(88.20%)、沙门氏菌(Salmonella)(5.49%)和葡萄球菌(Staphylococcus)(5.19%)。SEED 亚系统命中主要是结构部分,KEGG Orthology 命中与核苷酸和碳水化合物代谢相关的基因。未检测到抗微生物耐药基因。我们的噬菌体 DNA 纯化方法有利于噬菌体宏基因组学研究。柯斯噬菌体的优势可能解释了短尾噬菌体的罕见性。结构基因和辅助基因的优势可能是由于我们样本中溶菌噬菌体的丰度较高。即使在医院环境的噬菌体中也不存在抗生素耐药基因,表明噬菌体不是耐药基因的重要载体。