Metagenomic profiling of fresh water lakes at different altitudes in Pakistan

Freshwater lakes play a crucial role in sustainability of ecosystem in the favour of communities flourishing around it. In this study, differential metagenomic evaluation of three freshwater ecosystems in Pakistan was carried out. These included Lakes Keenjhar, Rawal and Saif-ul-Muluk situated at the altitudes 21, 527, 3224 m respectively. Former two lakes serve as water reservoirs for three major cities of Pakistan, while the high-altitude Lake Saif-ul-Muluk is located in a pristine environment at the meeting point of Himalayan and Karakoram ranges. Next generation sequencing (NGS) of metagenomic DNA of microbial communities was carried out using surface water samples from these lakes by Illumina Hi-Seq 2500 sequencing technology. Bioinformatics analysis identified Proteobacteria as the most dominant phylum (58–79%) followed by Planctomycetes (34%), Cyanobacteria (12%) and Bacteroidetes (15%). Lake Saif-ul-Muluk contained highest abundance of bacterial genera including Vibrio, Bordetella, Pseudomonas, Burkholderia and Escherichia; while Microcystis were found to be abundant in Lakes Keenjhar and Rawal. Non-parametric multi-dimensional scaling (NMDS) principle component analysis on genus level revealed that these ecosystems were divergent from each other and from metagenomes of lakes in the Americas. Characterization of virulence factors and antibiotics resistant genes in metagenomic contigs were also identified. Chemical profiling of Lakes Keenjhar and Rawal showed higher concentration of phosphorus which indicated eutrophication in these lakes might be due to Microcystis infestation.