This project develops plasmid and phage systems for the genetic manipulation of microbes in the built environment. These tools will allow 1) upregulation/downregulation/knockout of individual genes within built environment microbes, and 2) delivery of genetic material to specific microbes in situ, paving the way for functional studies. As a representative and important “built environment” habitat, this project is focusing on premise plumbing, including sinks, drains, and toilets.

Traditionally, waterborne disease outbreaks have been prevented through centralized water treatment in utility water treatment plants. However, several CDC studies have shown that a significant number of waterborne disease outbreaks result from opportunistic pathogens that reside in premise plumbing environments as opposed to those associated with either water treatment plants or water supplies. This project will identify and characterize opportunistic pathogens in hospital and home premise plumbing systems. The project also considers how the environment as well as an individual’s oral and skin microbiome may contribute to the development of premise plumbing biofilms and investigates potential exposure pathways.

Premise plumbing comprises the complete hot and cold water systems in a building and includes everything from the hot water heater and HVAC to the showers, faucets, sinks, and toilets. These systems are composed of a variety of materials (e.g. copper, PVC, PEX) that result in an environment that varies both temporally and spatially in its physical and chemical properties. This project collects fine scale data on the materials commonly used in premise plumbing and uses it to create a set of known substrates of varying properties that can be used to grow and characterize biofilms under varying environmental conditions.

A fundamental challenge in microbiome science is to understand the mechanisms that determine fitness of individual community members in a given habitat. The objective of this project is to establish multiple complementary approaches to defining genes and traits that confer fitness to members of built environment microbiomes. Planned methods include transposon insertion, heterologous expression, and high-throughput sequencing.