Telomeres are short and repetitive sequences at the ends of linear chromosomes which shorten with every cell division in vitro. Telomere length (TL) is reported to decrease with age and stress. The domesticated water buffalo (Bubalus bubalis) is the second important milk-producing animal worldwide. The productive lifespan of water buffalo cows is reported to be longer than that of dairy cows (Bos taurus). With this background, we aimed to compare in TL in blood leukocytes obtained from water buffalos across different ages. In addition, we tested the suitability of assessing TL in DNA derived from nasal and vaginal swabs as potential non-invasive alternatives for blood telomeres. The respective samples were collected from 20 calves (3 mo of age), 20 heifers (2 years old), 20 cows (first lactation, 3 years old age), and 13 cows (third lactation, about 5 years old) and TL was assessed by qPCR with β-globin as reference. One-way ANOVA with Bonferroni post-hoc tests was used to test for differences between the age groups. Pearson correlations were calculated to assess associations of TL obtained in different sample matrices (blood versus smears). The TL in blood leukocytes from water buffalo calves, heifers, and from cows in their first lactation was not different, but shorter telomeres were observed in cows in their third lactation. The results thus support an age-dependent decrease of TL in water buffalos. The TL recorded in leukocytes were weakly correlated with TL measured in DNA from nasal swabs (r = 0.327; P = 0.025), but not with TL from vaginal swabs. The weak and absent correlation of TL in nasal vaginal and nasal epithelial cells, respectively, and the unease of collecting nasal swabs from buffaloes, incapacitated these swabs as suitable alternatives for blood cells when assessing TL.