Christopher Sinigalliano

2015

2016

Both coastal inlets and treated wastewater outfalls are recognized as major pathways for microbial contaminants from Land-Based Sources of Pollution (LBSP) to enter Florida’s Southeastern coastal marine ecosystems (CRCP FY12 RFP, p.15), including an area containing reef tracks with an estimated annual economic value of $3.88 Billion (Johns et al, 2001). These LBSP discharges can contain a variety of microbial contaminants including fecal indicator bacteria, fecal host marker bacteria, and pathogens to both human health and coral ecosystem health. Previous studies by AOML have shown that both coastal inlets and treated wastewater outfalls can be a substantial source of microbial particulates and bacterial genetic material from LBSP discharges, although bacteria from treated wastewater outfalls were predominantly dead or dormant, whereas bacteria from inlets were predominantly live and potentially infectious. Previous studies by our laboratory with the NOAA FACE program have measured specific LBSP-associated bacterial contaminants discharged from the Boynton Inlet, Hillsboro Inlet, and Port Everglades Inlet using both culture-based methods and molecular microbial source tracking methods. These microbial contaminants have also been measured in surface reef waters from Broward to Palm Beach. Previous microbial source tracking studies have shown that these LBSP microbial contaminants can at least periodically be transported to coral reef waters of the region (Carsey et al 2011). However, these prior FACE projects did not provide an opportunity to actually sample coral nor to determine or measure the extent to which such microbial contaminants can incorporate into the bacterial populations of actual coral reef mucus or tissue. This represents a significant information gap. If such microbial contaminants are impacting corals, then molecular source tracking conducted on coral samples may help identify originating host sources, and help direct mitigation efforts. Likewise, quantification/characterization of coral symbiont communities can provide a diagnostic for coral health and bleaching far in advance of other metrics. The need to discriminate LBSP sources of contamination has been articulated by managers and by the 2010-2015 CRCP National Goals/Objectives. There is an opportunity to leverage against ongoing AOML field programs which will conduct bimonthly water quality cruises over FY 14 and FY 15 to sample waters of Port of Miami Inlet, Bakers Haulover Inlet, and Port Everglades Inlet, the two Miami area treated oceanic wastewater outfalls, and four regional coral reef sites potentially impacted by LBSP discharge. This field program will also include biometric surveys by divers of 3 coral sites for each of 4 regional reefs. However this field program does not currently have a microbiology component. We propose here to leverage against this field program to collected bimonthly water samples and quarterly coral samples for molecular microbial analysis of LBSP bacterial contaminants by both target specific qPCR methods and by total microbial community next-generation sequencing and metagenomic analysis, and to characterize coral symbiont populations by qPCR. Correlating patterns of pollutant exposure with microbial community composition may identify threshold levels of exposure, and PCR source tracking may identify originating host sources to discriminate between sewage/septic, agricultural, urban runoff, etc., thus aiding in targeted mitigation efforts.

This project is expected to accomplish the following: (1) Bimonthly microbial measurements from water quality cruises over two years for sites from 3 coastal inlets, 2 treated wastewater outfalls, and 4 coral reefs. Microbial measurements will include viable fecal enterococci plate counts, and the following microbial source tracking markers and pathogens by qPCR: total enterococci, total Bacteroidales, human-source Bacteroidales, dog-source Bacteroidales, cow-source Bacteroidales, gull/seabird-source Catellicoccus, Escherichia coli, Serratia marcescens (causative agent of coral white pox), and Staphylococcus aureus. (2) Quarterly microbial measurements of coral mucus and tissue for 3 different sites each from the 4 reefs for the same microbial source tracking markers and pathogens, plus Symbiodinum community characterization by qPCR. (3) Quarterly characterization of total microbial community structure in the waters of the 3 inlets, 2 outfalls, and in the tissues of the 12 coral sites described above, using Illumina-sequencing and community metagenomic analysis to compare microbial community structure between inlets, outfalls, and coral microbial populations. The community sequence data will also be screened for additional pathogens that might impact reef health besides the ones specifically targeted by qPCR. This work will help inform management as to what degree LBSP bacterial contaminants may be influencing coral reef microbial communities, how coral symbiont populations may be reacting to LBSP exposure, and help discriminate the potential LBSP sources for these contaminants, such as sewage/septic, agricultural runoff/canal discharge, urban runoff, seabird fecal deposition, etc., which may better direct management and mitigation resources and efforts, thus improving both reef and public health.

• Florida

• N/A

Land-based Sources of Pollution (LBSP)

Closed

Completed

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