Canis familiaris is a domestic animal which plays an important role as a pet; however, it is very likely to develop diseases characterized by high morbidity and mortality. In this sense, there are some Ancylostomatidae species that can lead to anemia, weight loss, and abdominal pain. Therefore, the treatment with allopathic drugs is essential for eliminating the parasitism. However, the increasing occurrence of resistance to allopathic treatments stimulates the search for new therapeutic inputs, among which the probiotics stand out and are able to positively alter the intestinal microbiota and exert immunomodulatory effect. Therefore, the present study aimed to evaluate the applicability of using species of Lactobacillus ( L. acidophilus ATCC 4536, L. plantarum ATCC 8014, and L. delbrueckii UFV H2B20) to control canine ancylostomiasis. A probiotic preparation containing 1 × 10 6 CFU of each strain was administered to 10 naturally infected animals, on alternate days for 28 days. It was observed that the treatment with the probiotic preparation led to a significant reduction in EPG of feces (88.83%/ P < 0.05), as well as an increase in leukocyte and lymphocyte counts, highlighting the potential use of Lactobacillus species in the control of canine ancylostomiasis.