Antibiotic resistance in the environment occurs naturally in bacteria, and is a growing problem due to the use of antibiotics both in human and animal medicine. β-lactamases are the most studied group of antibiotic resistant genes, as these enzymes hydrolyze penicillin, ampicillin, and other β-lactam antibiotics. The purpose of this study was to determine if there was a statistically significant difference between numbers of copies of β-lactamase genes (bla1, bla-TEM, bla-SHV) from ampicillin-resistant bacteria in soil samples from a horse pasture and a crop field. Colony counts suggested there was a significant difference (p < 0.05) in ampicillin-resistant bacteria between the two soil samples. Upon completion of PCR and gel electrophoresis, there was no statistically significant difference in the presence of bla1 and bla-TEM (p > 0.05) in bacteria from each soil sample, but there was a significant difference in the presence of bla-SHV (p = 0.0061). A significant difference in ampicillin-resistant colonies and bacteria with bla-SHV suggests that ampicillin used in the horses may be selecting for ampicillin resistance in the soil bacteria. There may also be high concentrations of other ampicillin-resistant genes present in agricultural rich soils, particularly in animal pastures. Future studies need to be undertaken to document the level of antibiotic resistance found in animal pastures to understand the impact this may have on animal and human health.