Background: The widespread indiscriminate application of antibiotics to food crops to control plant disease represents a potential human health risk. In this study, the presence of antibiotic-resistant staphylococci associated with workers and orange orchard environments was determined. A total of 20 orchards (orange and other fruits) were enrolled in the study. Trees in the orange orchards were treated with ampicillin on a pre-determined schedule. Environmental samples (n = 60) included soil, water, and oranges; 152 hand and nasal samples were collected from 76 healthy workers. Antibiotic susceptibility profiles were determined for all staphylococcal isolates. Results: This investigation revealed that of the total Staphylococcus spp. recovered from the orange orchard, 30% (3/10) were resistant to erythromycin, 20% (2/10) were resistant to ampicillin, and 20% (2/10) resistant to both erythromycin and ampicillin. Conclusion: The application of antibiotics to orange trees in open production environments to halt the spread of bacterial disease presents risks to the environment and creates health concerns for Thai farmers using those agents. ARB on crops such as oranges may enter the global food supply and adversely affect public health.