Lactic acid bacteria (LAB) are related to antimicrobial activity, improve animal health and productivity, protect against some intestinal infections, present anti-inflammatory properties, and improve immune status. The objective of this study was to evaluate the probiotic capacity of LAB isolated from the guinea pig (Cavia porcellus) intestine. The antibacterial capacity of LAB (molecularly identified) against Salmonella typhimurium, Klebsiella pneumoniae, Escherichia coli, and Staphylococcus aureus was evaluated. Exopolysaccharide production, adherence capacity, and antibiotic resistance of LAB were measured. The effects of pH (2.9, 5.0, 6.4, and 7.4), temperature (40, 50, and 60°C), and different NaCl concentrations (3.5 and 6.5% w/v), on LAB growth, were evaluated. Strains of the genera Ligilactobacillus, Weissella, Enterococcus, and Limosilactobacillus were molecularly identified. Ligilactobacillus salivarius strains (ACI1, GCI9, and GDE10) had the highest adherence capacity. L. salivarius strains (ACI1 and GCI9) produced the most exopolysaccharides (1.71 and 1.76 mg/mL, respectively), and were the best adapted to different pH and temperatures. Additionally, the strains of L. salivarius (GCI9 and GDE10) isolated from the cecum and intestine of guinea pigs could be used for the prevention or control of pathogen infections in guinea pig farms, due to their adherence capacity and antimicrobial activity against Salmonella typhimurium, Klebsiella pneumoniae, Escherichia coli and Staphylococcus aureus, pathogens responsible for individual or mixed bacterial infections of the digestive, respiratory and urinary systems in guinea pigs, which would reduce morbidity and mortality in this species that is part of the traditional gastronomy in South America. In addition, its high tolerance to temperatures up to 50°C makes this strain ideal for the development of food matrices directed to minor species such as guinea pigs.