Dave Bachoon

1. Evaluating the Presence of Pathogenic Bacteria in Fecal Samples of
Feral Pigs and Their Transport to Surface Water
Dave Bachoon, Jyoti Lama, Brennan Pookwong,
Biology & Environmental Sciences, Georgia College & State University, Georgia
Abstract Methods
Results
Acknowledgements
References
• The primary goal for this project was to improve understanding of
pathogenic bacteria harbored by feral pigs and their impact on water
quality. Fecal samples of feral pigs from five counties in Georgia were
collected and the DNA was extracted. qPCR screening of fecal
samples for the presence of Brucella suis, Campylobacter jejuni, and E.
coli O157:H7 indicated that B. suis was present in 25% of the feral pigs
and E. coli O157:H7 was present in 5.5% of the pigs but Campylobacter
jejuni was not detected in any of the feral pig fecal samples. The
majority of majority of pigs carrying zoonotic pathogens came from
Morgan County. Phylotyping analysis of E. coli isolates from the feral
pigs indicated that these pigs carry a high proportion of virulent strains of
E. coli (phylogroups B2 and D) compared to cattle and farm raised pigs.
DNA samples are being processed for metagenomic analysis of Brucella
and Escherichia groups. Water samples are being collected for analysis
of pathogenic bacteria and the presence of fecal bacteria from feral pigs.
This project gives Georgia College, a small university steadily building
its research portfolio, a valuable opportunity for expanding the
institution’s research capabilities and funding potential.
Results
dave.bachoon@gcsu.edu
• Figure 2. Feral pigs on farmlands. Photos provided by M.
Ondovchik USDA-APHIS, Athens Ga.
• This project is funded through The National Institute of Food and Agriculture
(NIFA), USDA. Sustaining water quantity, quality, and availability for agricultural
use while maintaining environmental quality through 2050.
(2015-68007-23137).
• We appreciate the collaboration of Dr. Marirosa Molina at USEPA in Athens,
GA.
• We appreciate the assistance of Rob Pinkston of Yager Pro and USDA-APHIS
in Athens for help with sample.
Conclusion
Introduction
Specific objectives: Summer sabbatical
1. To evaluate the presence of pathogenic bacteria in fecal samples from feral
pigs on cattle farms in relation BMP’s and transport to surface waters.
2. Develop my expertise in metagenomic analysis of bacterial population of
fecal samples from feral pigs.
Trapped feral pigs
Figure 3. Water and fecal sampling sites in areas where feral
pigs are active. BC, Beef cattle, DC, dairy cattle, NC, no cattle.
Table 1. QPCR and MST primers used
Grant number: 2015-68007-23137
Figure 3. Dichotomous decision tree to determine the phylogenetic group of an E. coli
strain (Clermont et al., 2000).
Dick, L. K., and Field, K. G. (2004). Rapid Estimation of Numbers of Fecal Bacteroidetes by Use of a
Quantitative PCR Assay for 16S rRNA Genes. Applied and Environmental Microbiology 70(9),
5695-5697.
Guion, Chase E., Theresa J. Ochoa, Christopher M. Walker, Francesca Barletta, and Thomas G.
Cleary. "Detection of diarrheagenic Escherichia coli by use of melting-curve analysis and real-time
multiplex PCR." Journal of clinical microbiology 46, no. 5 (2008): 1752-1757.
Haugland, R.A., Varma, M., Sivaganesan, M., Kelty, C., Peed, L., and Shanks, O.C., (2010).
Evaluation of genetic markers from the 16S rRNA V2 region for use in quantitative detection of
selected Bacteroidales species in human fecal waste by qPCR. Systematic and Applied
Microbiology, 22, 348-357
B.J. Kildare, C.M. Leutenegger, B.S. McSwain, D.G. Bambic, V.B. Rajal and S. Wuertz, 16S r RNA-
based assays for quantitative detection of universal, human-, cow-, and dog-specific fecal
Bacteroidales: a Bayesian approach, Water Res. 41 (16) (2007), pp. 3701–3715
Mieszkin, S., Furet, J. P., Corthier, G., and Gourmelon, M. (2009). Estimation of Pig Fecal
Contamination in a River Catchment by Real-Time PCR Using Two Pig-Specific Bacteroidales 16S
rRNA Genetic Markers. Applied and Environmental Microbiology 75(10), 3045-3054.
• qPCR screening of fecal samples for the presence of Brucella suis,
Campylobacter jejuni, and E. coli O157:H7 indicated that B. suis
was present in 25% of the feral pigs and E. coli O157:H7 was
present in 5.5% of the pigs.
• Campylobacter jejuni was not detected in any of the feral pig fecal
samples.
• Phylotyping analysis of E. coli isolates from the feral pigs indicated
that these pigs carry a high proportion of virulent strains of E. coli.
• With the increasing numbers of feral pigs in the country, more
studies are warranted on the diversity of potential pathogenic
microbes carried by feral pigs.
Figure 1—In 1998, feral swine could be found in parts of all the counties shown here in green. By
2004, wild pigs had increased their range to counties shown in yellow, while remaining in the
green-colored areas as well. Note that wild pigs are now found in two counties in Iowa—America’s
#1 swine-producing State. (Map adapted from originals created by the Southeastern Cooperative
Wildlife Disease Study, Athens, GA.)