Though world buffalo population is about 1/9 of cattle population, more human beings depend from buffaloes, especially in South-East Asia. Indeed, there are about 168 million water buffaloes in the world, mostly (161 million) raised in Asia. The River buffalo has received great attention from West countries which are particularly interested to both milk and meat production. For this reason, the genetic improvement of buffaloes still remains one of the most important goal in this species. Cytogenetic is one of the biotechnologies which supports the genetic improvement of buffaloes, especially for the selection of reproducers. In this chapter, an update of the latest results obtained in the cytogenetics of buffaloes is reported, starting from its cytotaxonomy and going through clinical and molecular cytogenetics, cytogenetic investigations and breeding objectives.