May 2009

Beating Bacteria

A new methodology for identifying and prioritizing water bodies with high concentrations of fecal coliform is helping Florida address the problem of elevated bacteria levels.

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Photo: Valerie J. Harwood

By Cheryl M. Wapnick, Thomas L. Singleton, Valerie J. Harwood

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Pertinent information related to the watershed in question is acquired and compiled via the contaminant source survey (CSS), which is a multifaceted approach designed to identify the most probable bacterial sources. In keeping with the phased-monitoring concept recommended by the NRC (2004), the intensity of an investigation—that is, a CSS—at a given site is based on its MWQA classification. Development of a CSS includes detailed reviews of existing data, including historical sampling results and geographic information system (GIS) data. Collaboration with local stakeholders occurs in the form of intensive one-on-one interviews, as well as public workshops addressing local water-quality issues and potential sources of fecal contamination. Field reconnaissance, and, if necessary, advanced microbial source tracking (MST) (step 3), also are used. For example, sites in MWQA categories D and E are slated for more intensive investigations, including MST sampling programs, than sites exhibiting less-frequent (and lower-magnitude) exceedances of bacterial standards. The relatively large expense associated with MST analyses emphasizes the importance of using available data as effectively as possible.

During field reconnaissance, probable sources and migration pathways are assessed, and potential sampling stations are identified. Participants also develop a sampling implementation plan. Once the study is completed and contamination sources are identified, appropriate actions for addressing the bacterial problem can be developed and initiated. Because local stakeholders participate in all aspects of the CSS, the results reflect a partnership focused on identifying the most probable sources.

Step 3: Microbial Source Tracking
As the most technically advanced component of the CSS, MST is typically conducted in those watersheds with the highest frequency and magnitude of exceedance (MWQA categories D and E). Specific MST tests for various types of human and/or animal contributors to contamination are chosen according to a decision-tree approach. To reduce costs and time, the sampling begins by using less expensive, more basic analytic methods first, including the combined use of different indicator bacteria (for example, fecal coliforms, E. coli, and enterococci), followed by more costly and sophisticated methods, including source-specific assays. Using multiple MST methods increases the confidence in source identification and expands the range of potential sources that can be investigated.

MST methods are used to detect a specific gene or genes found in a microorganism that is specific to a certain host or group of hosts. Polymerase chain reaction (PCR) is frequently used to detect the target gene(s). PCR is a process in which many copies of a specific DNA sequence are synthesized from a small amount of starting material, generating enough material to be visualized or manipulated for further investigation. The target genes may be present in fecal indicator bacteria, microbial pathogens, viruses, or even host cells that are shed in feces (Stoeckel and Harwood 2007).

Working with laboratories throughout Florida, particularly at USF, PBS&J employs a “toolbox” of MST methods that currently includes non-library-based PCR tests. One of the targets is a group called the Bacteroidales, which are anaerobic fecal bacteria. The DNA of some members of this group lends itself to three separate tests for detecting whether a source is human-, ruminant-, or horse-specific. Other non-library-based PCR and qualitative PCR tests use the Enterococcus faecium esp gene, a gene for virulence factor of a bacterium found in humans, and the human polyomavirus, a nonpathogenic virus shed in urine and, therefore, commonly found in sewage. As new cost-effective MST methods, including quantitative PCR, are developed, they are added to the toolbox.

Photo: PBS&J
Contaminated water from a small tributary to the Lower Hillsborough River flows downstream, eventually entering the main channel of the river.
Findings from the Decision-Support Tool can be used by the FDEP or local stakeholders to determine baseline conditions before implementing corrective actions to address sources of contamination. The results also help to focus an investigation, providing a framework for interpreting and responding to bacterial exceedances. Finally, the results of the Decision-Support Tool can be easily communicated by means of a simple “stoplight” approach that ranks sites based on the apparent risk that they pose to human health (Table 1).

Applying the Decision-Support Tool to the Lower Hillsborough River
Working with FDEP and local stakeholders in the Hillsborough River watershed, PBS&J employed the overall methodology to help guide implementation of a BMAP for several water bodies within the area that were verified as impaired as a result of elevated fecal coliform concentrations.

Although the effort involved an examination of six sub-basins, only the Lower Hillsborough River sub-basin is discussed here. Monitoring locations along the water body—a highly urbanized waterway that experiences significant recreational use—were classified using the Decision-Support Tool. The locations received MWQA classifications on the basis of long-term monitoring data collected by the Hillsborough County Environmental Protection Commission (EPCHC) as well as more recent MST data that tested for markers specific to humans, ruminants, and horses. Human-specific markers were found at almost every site tested. These results often coincided with exceedances of indicator organisms. Ruminant sources, by contrast, appeared to be present but at relatively insignificant levels.

Photo: PBS&J
Station DHR4A, which is located on the tributary, was found to be the most impaired site during the application of the Decision-Support Tool within the basin.
The first stage of the CSS, which was conducted before the MST testing, resulted in the identification of potential sources of fecal contamination and a thorough understanding of the characteristics of the Lower Hillsborough River. For example, several minor tributaries, spring-fed drains, and significant stormwater conveyance systems that join the main channel were identified. The most significant potential source of fecal contamination was determined to be human-associated, including impacts from the sewer system and associated large-scale and repetitive sanitary sewer overflows, septic systems, homeless populations, and live-aboard vessels docked at marinas along the river. The potential contribution of non-human-related impacts was found to be comparatively low, including stormwater, bird populations and other wildlife, dogs, and animals housed at the local zoo. Recognition of these potential sources of impairment and their locations played a critical role in designing the MST portion of the assessment, resulting in the use of lower-cost, more basic methods first, followed by higher-cost, more sophisticated methods. In this way, the costs and time associated with the MST were kept to a minimum.

MST results showed that human-related contamination was indeed the leading cause of fecal pollution identified within the Lower Hillsborough River. More importantly, the sampling program resulted in the ability to define the most probable sources for individual locations throughout the watershed (Figure 3). This allowed participants to effectively target additional investigation efforts—for example, infrastructure evaluations—and the required management actions to address the specific type of contamination at each particular site. Furthermore, sampling of the sediments at several sites throughout the basin showed high indicator bacteria levels, signifying periodic inputs or extended persistence of indicator bacteria. Each of these locations is suspected of being subject to recent or past wastewater leaks that may have contributed to significant reservoirs of bacteria in the sediments throughout the Lower Hillsborough River. These reservoirs have the potential to re-inoculate the water column and contribute to chronic and elevated levels of surface water contamination.

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The frequency and severity of indicator bacteria exceedances identified during the sampling aspect of the methodology prompted additional investigations by local stakeholders. For example, the city of Tampa, the Hillsborough County Health Department, and the EPCHC all made multiple visits to station DHR4A, which is located on a small tributary to the Lower Hillsborough River where human-specific markers and extremely elevated indicator bacteria counts were routinely detected. The collaborative efforts have determined that there are likely two separate sources that combine just upstream of station DHR4A. Local stakeholder efforts are currently underway to identify the exact sources of the contamination in this area.

Certain corrective actions were recommended as a result of the findings from the methodology. For example, predictive and preventative maintenance programs for wastewater infrastructure were suggested. Such programs are intended to identify problematic areas and determine the “root causes” of failures involving sewers and pump stations. Regular inspection and cleaning practices were also advised, along with efforts to address problems relating to fats, oils, and grease. To ensure greater accountability relating to private infrastructure, a comprehensive GIS database of such systems is to be developed. Next Page >

What Do You Think?

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bullardrr

May 6th, 2009 12:13 PM PT

For a great number of watersheds, the spectrum and quantities of microbial indicators may be highly episodic, significantly associated with major rain run-off events, in which case the grab samples are best timed to correspond around the time of concentration for the location of the sampling station. At other sites a rote interval of collection may accurately portray the situation.

ldwaters

April 16th, 2009 8:48 AM PT

It would be helpful to have a case study that showed how the decision-support tool method fared when neither humans nor other animals for which non-library methods of genetic identification are the source of fecal coliform, such as rodents, raccoons, etc. How does one use microbial source tracking to identify plant-based sources of fecal coliform bacteria, such as decomposing citrus or waste paper? More detail on the field survey part is needed since this portion of the tool will be the most informative in those cases where microbiology yields inconclusive results. L. D. Waters

edo

April 8th, 2009 11:23 AM PT

This is a highly useful paper and by its publication it has considerably boosted my respect for this journal. We need much more of this type of information in this profession. Dr Edo McGowan

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