Research in Greg Kleinheinz's Lab
Dr. Greg Kleinheinz’s interdisciplinary research group includes
over 12 student researchers (including both graduate and undergraduate)
working a variety of projects. Many students involved in Dr. Kleinheinz’s
research have served as both primary and secondary authors on numerous
professional presentations and publications. One of the most active
areas of research for Dr. Kleinheinz’s research group is the biology
of microbial indicator organisms present in recreational waters and
their relationship to public health. Traditionally, E. coli and/or
enterococci have been used as indicators of fecal contamination; however,
researchers have come to realize limitations regarding use of such indicators.
The limitations are no more apparent than when trying to determine the
source of the fecal contamination using these indicators. Thus, Kleinheinz’s
research group is exploring the sources of these indicator organisms
using antimicrobial resistance patterns, serological differences, and
the genetic uniqueness of each source of E. coli isolate. Understanding
the E. coli source is critical to understanding the risk posed
to human health by a certain quantity of these organisms when found
in recreational waters. As mentioned previously, E. coli is very
limited in its usefulness as an indicator organism, thus basic biological
aspects of novel indicator organisms need to be evaluated. One such
potential indicator that has not been researched in detail is the use
of the genus Bacteroides as an indicator of fecal contamination.
While there are some human-specific strains of Bacteroides, no
basic research has investigated the possible existence of an avian-
or bovine-specific strain to determine if Bacteroides could be
used as a reliable indicator of fecal contamination in water, as well
as tracking the source of the contamination.
After the conclusion of the essential proteomics and functional genomics techniques course, REU students will participate in a one-week field experience at one of Kleinheinz’s remote laboratories (Ashland, Eagle River, or Sturgeon Bay, WI). During this field experience the students will learn sample collection and handling techniques and will collect the field samples needed during the laboratory phase of the project. Once field samples are collected, the students will return to the laboratory to recover Bacteroides isolates from these samples. Once the isolates have been recovered, they will be catalogued. Genetic, biochemical, and morphological evaluation will commence. Each student will examine one or more techniques to evaluate each isolate. Specifically, isolates will be evaluated using tools contained within the NSF Proteomics and Functional Genomics Core Facility at UW Oshkosh. For example, students will obtain genetic fingerprints (using repetitive element PCR) and analyze them using advanced gel-imaging systems and software contained within the Core Facility. As an additional method of evaluation, students will investigate characterization of each isolate using MALDI mass spectrometry as has been previously described for other microorganisms. Once each isolate is fully evaluated, comparisons of the isolates within groups (i.e., human isolate versus human isolate will be conducted), as well as comparisons between groups of isolates (i.e., human versus seagull) will be made. Ultimately, students will have libraries of Bacteroides isolates from a variety of sources that will impart to them a comprehensive understanding of the uniqueness, or lack thereof, of each group of isolates. This work will serve as a foundation of basic knowledge essential to future applied studies looking at specific quantities of Bacteroides versus gastrointestinal disease of users of recreational waters and will serve as the first evaluation of quantity and diversity of Bacteroides isolates in recreational waters.
