Metagenomic Investigation of the Microbial Community Structure and Diversity for Sentinel Coral Reefs and Urbanized Coastal Waters in Southeast Florida, and Molecular Microbial Source Tracking to Characterize Potential LBSP Microbial Contaminant Influences

Coral reefs and recreational beaches provide critical ecosystem services. However, coastal waters of the Southeast Florida region receive anthropogenic discharges from highly urbanized watersheds via runoff, canals, coastal inlets, and treated wastewater outfalls. There is concern regarding the biological contaminants that enter the coastal zone from land-based sources, especially for viable pathogens and genetic elements that could confer virulence or resistance. Targeted molecular microbial source tracking (MST) by quantitative PCR allows the measurement of specific microbial contaminants such as host-specific fecal indicators. These fecal source markers can help track specific fecal contamination of public health concern in the coastal zone and may also help track exposure of coral reefs to such contamination. A range of pathogens associated with sewage/septic contamination have shown detrimental impact to coral communities, including changes to the biodiversity of coral microbiomes. High-throughput Next-Generation-Sequencing (NGS) and community genomic analysis can provide a comprehensive, culture-independent approach to investigate microbial community diversity in complex environmental samples. The combination of host-specific microbial source tracking by qPCR and metagenomic NGS can provide substantial enhancement to traditional methods of water quality assessment to better protect both environmental biodiversity and human health.

Reported here is a multifaceted water quality assessment study of three coastal inlets, two treated wastewater outfalls, and four sentinel coral reef communities in the Southeast Florida coastal zone offshore of Miami-Dade and Broward Counties. This study utilized a combination of bi-monthly sampling for nutrients, fecal indicator bacteria, and human-source molecular source tracking to measure specific contaminants of ecosystem and public health concern. In addition, 16S metagenomic analysis using Illumina Next-Generation Sequencing was used to establish a record of the microbial diversity from inlets, municipal wastewater outfalls, surface waters, and coral reef microbiomes, and to screen the community sequencing data for detection of additional pathogens not specifically targeted by the other methods.