Within the general aim of developing a Welfare Quality system for monitoring dairy buffalo welfare, this study focused on prevalence and interobserver reliability of the animal-related variables to be included in the scheme. As most of the measures were developed for cattle, the study also aimed to verify their prevalence for buffaloes. Thirty animal-based measures (22 clinical and 8 behavioral measurements) and 20 terms used for qualitative behavior assessment were assessed in 42 loose-housed buffalo farms. All farms were located in central-southern Italy. Two assessors were used (1 male and 1 female). The time needed to record all measures (animal-, resource-, and management-based) was 5.47 ± 0.48 h (mean ± SD). Interobserver reliability of animal-based measures was evaluated using Spearman rank correlation coefficient test (rs). If 0.7 is considered as threshold for high interobserver reliability, all animal-based measures were above this level. In particular, most of the coefficients were above 0.85, with higher values observed for prevalence of animals that can be touched (rs = 0.99) and prevalence of animals with iatrogenic abscess (rs = 0.97), whereas lower coefficients were found for the prevalence of vulvar discharge (rs = 0.74) and dewlap edema (rs = 0.73). Twelve out of the 20 terms used for the qualitative behavior assessment reached a satisfactory interobserver reliability (rs = 0.65). Principal component analysis of qualitative behavior assessment scores was conducted for each assessor. Both principal component 1 and principal component 2 showed high interobserver reliability (rs = 0.80 and 0.79, respectively). In addition, relevant proportions of animals were affected by welfare issues specific to buffaloes, such as overgrown claws (median = 34.1%), withers hygroma (median = 13.3%), and vulvar or uterine prolapse (median = 9.3%). We concluded that most of the investigated measures could be reliably included in the final scheme, which can be used as such to monitor buffalo welfare. However, to inform consumers about the welfare status of the animals, the data should be integrated into a single overall assessment of animal welfare, as already performed in the Welfare Quality project for dairy cattle.