The zoonotic potential of Cryptosporidium parvum was studied in an extensive cattle farming region of northern Tunisia. Seventy fecal samples from pre-weaning calves and 403 fecal samples from children were examined by microscopy after modified Ziehl-Neelsen (MZN) staining. Positive Cryptosporidium specimens were identified at a species level using an 18S rRNA nested polymerase chain reaction (PCR) followed by an Restriction Fragment Length Polymorphism (RFLP) analysis. C. parvum isolates were subgenotyped by sequence analysis of the glycoprotein 60 (gp60) gene. Among calf samples, 14 samples were positive by MZN method. C. parvum was identified in all cases. Twelve parvum isolates (85.7%) belonged to family subtype IIa. Subtype IIaA15G2R1 was more prevalent (50%). Two C. parvum isolates corresponded to the IIdA16G1 subtype. Seven human samples were positive by MZN method. C. parvum and C. meleagridis were identified in four and three cases, respectively. Intraspecific characterization of C. parvum identified two subtypes, the IIaA15G2R1 and the IIdA16G1, also found in calves.