The aims of the presented study were (1) to examine the relationship between foetal measurements and gestational age (GA), (2) to generate GA formulas, and (3) to investigate the estimation of GA by transabdominal ultrasonography in buffaloes. Thirteen pregnant buffaloes were used in the study. Transrectal ultrasonography was performed between Day (D) 28 and 112 of gestational age, whereas transabdominal ultrasonography was between 126 and 294. The diameters of embryonic (EVD) and amniotic (AVD) vesicles, crown-rump length (CRL), occipito-nasal length (ONL), biparietal diameter (BPD), orbit diameter (OD), cervical, thoracic, lumbar and coccygeal vertebrae lengths (CVL, TVL, LVL, CcVL), abdominal diameter (AD), chest diameter (CD), umbilical cord diameter (UCD), scapula, humerus, radius-ulna, metacarpus, femur, tibia, and metatarsus lengths (SL, HL, RUL, McL, FL, TL, MtL), diameters of transversal heart (THD), stomach (SD), kidney (KD), and outer diameter, circumference and area of placentomas (OPD, OPC, OPA) were measured by ultrasonography. All 26 parameters were highly correlated with GA (r = 0.968 - 0.999). The observation of the foetus was evident in all animals via transabdominal ultrasonography, and all parameters except EVD, AVD, and CRL could be measured on D 126. In addition, heartbeats, the sign of foetal vitality, could be observed in 11 of 13 living foetuses. This study is the first to demonstrate that transabdominal ultrasonography can be used to estimate GA in buffaloes. In addition, GA formulas related to ONL, CVL, TVL, LVL, CcVL, extremity bone lengths, THD, UCD, PC, and PA measurements were created for the first time in buffaloes.