Serum samples from 726 Ecuadorian children who underwent natural rotavirus (RV) exposure were tested for neutralizing serum antibodies against two serotype 6 (ST6) human RV (HRV) isolates from Italy, PA151 and PA169, and two ST6 bovine RV (BRV) isolates, NCDV and UK. Gene 4 was distinct in all four ST6 strains. Ninety-one, 56, 67, and 65 serum samples neutralized HRV PA151 (13%), HRV PA169 (8%), BRV NCDV (9%), and BRV UK (9%), respectively. A total of 44 of the 91 serum samples which neutralized HRV PA151 did not neutralize the other three ST6 RV strains. In addition, we identified three serum samples that neutralized HRV PA151 but none of the six human or four animal RV STs. However, we failed to identify serum samples that neutralized HRV PA169 without neutralizing at least one of the major HRV STs. With a hospital-based serum collection from German children (excluding gastroenteritis patients), we identified 3 out of 197 serum samples tested that neutralized HRV PA151 specifically, whereas none neutralized HRV PA169 exclusively. None of the 71 German infants hospitalized with primary RV gastroenteritis showed a PA151- or a PA169-specific antibody response.