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Differences in rumen volatile fatty acids (VFAs), microbial population and community composition between water buffalo and Jersey cows were compared to explore the effects of animal species on ruminal microbiota. Ruminally fistulated buffalo (n = 4) and Jersey cows (n = 4) with similar body weights were selected and provided with similar diets. After feeding for 30 days, rumen fluid was collected to quantify the ruminal VFA, and determine microbial population and community composition. Results revealed that concentrations of total VFA, acetate and propionate in buffalo were higher (P > .05), while butyrate concentration and acetate/propionate ratio were lower than those in Jersey cows (P < .05). Populations of ruminal bacteria, protozoa, fungi and F. succinogenes were higher in buffalo (P < .05). The abundance of Firmicutes was higher (P < .05), while the abundance of Bacteroidetes was lower in buffalo (P < .05). The abundances of dominant bacterial genus Prevotella and dominant methanogen genus Methanobrevibacter were higher in Jersey cows (P < .05). Protozoal genus Metadinium was dominant in buffalo, while Entodinium was dominant in Jersey cows. Concluded that not only rumen microbial population, but also microbial community composition were different between two species. The higher bacterial, protozoal and fungal populations, and different microbial community can explain why buffalo are more efficient to digest fibre. [ABSTRACT FROM AUTHOR]