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In recent years, there has been a more prudent use of drugs on livestock farms and alternative products have been considered, with a view of reducing the risk of the onset of antibiotic resistance. <i>Salmonella enterica</i> subsp. <i>enterica</i> serovar Enteritidis (<i>S.</i> Enteritidis) may cause disease in poultry, and it is also responsible for human food poisoning. The aim of this study was to evaluate the efficacy of garlic against <i>S.</i> Enteritidis and to define its Minimal Inhibitory Concentration (MIC)₉₀ and MIC₅₀ values. The study was carried out in vitro, testing 26 <i>S.</i> Enteritidis strains identified in laying hens from various farms in Southern Italy. A preliminary efficacy trial was carried out on two strains, <i>S.</i> Enteritidis and <i>Escherichia coli</i>, using a garlic concentration of 10 mg/mL (1%). Later, 26 strains of <i>S.</i> Enteritidis at 10⁶ Colony Forming Unit (CFU) and 10⁴ CFU were tested with different concentrations of garlic, ranging from 10 mg/mL to 1 mg/mL. Based on the results, intermediate concentrations of garlic, from 5 mg/mL to 4 mg/mL and 4 mg/mL to 3 mg/mL, were used to test 10⁶ CFU and 10⁴ CFU, respectively. The data were statistically analyzed. The MIC₉₀ was 4.75 mg/mL for strains tested at 10⁶ CFU and 4 mg/mL for strains tested at 10⁴ CFU. The results highlight garlic’s potential to inhibit the growth of <i>Salmonella enterica</i> ser. Enteritidis in vitro. Efficacy was dependent on the microbial concentration used. In vivo efficacy trials will be crucial to confirm the efficacy of garlic against <i>S.</i> Enteritidis and to assess whether garlic can be used in poultry flocks to prevent the spread of the bacterium in the field.