Assessing the microbial quality of a tropical watershed with an urbanization gradient using traditional and alternate fecal indicators Tasha M. Santiago-Rodriguez, Gary A. Toranzos and Javier A. Arce-Nazario ABSTRACT Urbanization affects the microbial loading into tropical streams, but its impact on water quality varies across watersheds. Rainfall in tropical environments also complicates microbial dynamics due to high seasonal and annual variations. Understanding the dynamics of fecal contamination in tropical surface waters may be further hindered by limitations from the utilization of traditional microbial indicators. We measured traditional (Enterococcus spp. and Escherichia coli), as well as alternate (enterophages and coliphages) indicators of fecal contamination in a tropical watershed in Puerto Rico during a 1-year period, and examined their relationship with rainfall events across an urbanization gradient. Enterococcus spp. and E. coli concentrations were 4 to 5 logs higher in non-urbanized or pristine sites when compared to enterophages and coliphages, suggesting that traditional fecal indicator bacteria may be natural inhabitants of pristine tropical waters. All of the tested indicators were positively correlated with rainfall and urbanization, except in the most urbanized sites, where rainfall may have had a dilution effect. The present study indicates that utilizing novel indicators of microbial water quality may improve the assessment of fecal contamination and pathogen risk for tropical watersheds. Tasha M. Santiago-Rodriguez Department of Biology, California Polytechnic State University, San Luis Obispo, CA, 93407, USA and Center for Applications in Biotechnology, California Polytechnic State University, San Luis Obispo, CA 93407, USA Gary A. Toranzos Department of Biology, University of Puerto Rico, San Juan PR 00932, Puerto Rico Javier A. Arce-Nazario (corresponding author) Department of Biology, University of Puerto Rico, Cayey PR 00736, Puerto Rico and Instituto de Investigaciones Interdisciplinarias, University of Puerto Rico, Cayey PR 00736, Puerto Rico E-mail: javier.arce@upr.edu Key words | coliphages, Enterococcus spp., enterophages, Escherichia coli, rainfall, urbanization INTRODUCTION The microbial quality of tropical watersheds is traditionally monitored using Enterococcus spp. and Escherichia coli. These two indicators have an extensive history of being used for water quality regulatory purposes under United States laws, especially the Safe Drinking Water Act (NRC ). While these bacteria are found in the gut of warm-blooded ani- mals and may indicate fecal contamination, they are also natural inhabitants of pristine waters and originate from non- fecal sources; thus, their detection may not always indicate fecal contamination (Fujioka et al. , ; Solo-Gabriele et al. ; Noble et al. ; Byappanahalli et al. ; Boehm & Sassoubre ). In addition, traditional indicator bacteria-culturing methods provide results at 1824 h and additional conrmation steps are required, preventing immedi- ate actions to be taken to prevent potential risks to public health (Wade et al. ; Noble & Weisberg ). For these reasons, alternate indicators of fecal contamination may exhibit a higher source-specicity and may be more cost- and time-efcient (Sinton et al. ; Griffin et al. ; Bonilla et al. ; Purnell et al. ; Kent et al. ; Rusinol et al. ). For instance, polymerase chain reaction (PCR)-based methods, such as quan- titative PCR (qPCR), are increasingly being utilized to determine the microbial quality of waters. PCR-based methods have several advantages, including high-specicity and the reduced time to obtain results (Wade et al. ), but there are signicant drawbacks. These still primarily target 796 © IWA Publishing 2016 Journal of Water and Health | 14.5 | 2016 doi: 10.2166/wh.2016.041 Downloaded from https://iwaponline.com/jwh/article-pdf/14/5/796/393935/jwh0140796.pdf by guest on 30 December 2018