The objective of this study was to investigate the effect of reduced fat content in buffalo milk on textural, chemical and microbiological characteristics of plain buffalo yoghurt during storage at 4 °C. Four yoghurt samples were manufactured with different fat ratios (1.5–6 g/100 g) and analysed on the first, 10th and 20th days of storage. Increasing the fat content of the yoghurt increased the L* value, viscosity, and cohesiveness and decreased the a* value, syneresis, hardness, and springiness. Fat reduction did not cause any significant changes in the adhesiveness, gumminess, titratable acidity, b* value, and Streptococcus thermophilus population of buffalo yoghurt samples. Storage time had no significant effect on adhesiveness, springiness, and L* value of buffalo yoghurts. A decrease in S. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus populations during storage was detected. It was concluded that more acceptable buffalo yoghurt could be produced from milk with 3 g/100 g fat.