Ticks constitute one of the major health challenges to livestock production in Nigeria. They are particularly important due to their blood-feeding activity causing anaemia and as vectors of pathogens causing many livestock diseases. Accurate identification of ticks infesting livestock is fundamental for effective control of tick and tick-borne diseases. We integrated morphological identification and DNA barcoding to unambiguously identify the species of ticks and assessed their prevalence on cattle and sheep in Nsukka, Nigeria. All the animals in the farm: 37 cattle and 22 sheep were examined. All the cattle examined (100%) and 9 (40.09%) of the sheep were infested with ticks. The ticks Amblyomma variegatum and Rhipicephalus microplus were found on cattle and sheep in Nsukka. A total of 317 ticks were collected out of which 272 were on cattle and 45 on sheep. Of the total number of ticks collected, 152 (47.95%) were A. variegatum while 165 (52.05%) were R. microplus. Tick infestation was significantly higher on cattle than sheep (p<0.05, χ 2 = 3.902, p = 0.0482, n=59). The barcode region of COI gene was successfully amplified and sequenced in all the samples of ticks analysed. Samples of A. variegatum were identified at 99.70-100%. Similarly, samples of R. microplus were identified at 100%. Phylogenetic analysis clustered A. variegatum and R. microplus with their respective species from GenBank on phylogenetic tree. The ticks exhibited morphological ambiguity and sexual dimorphism, but DNA barcoding resolved the identities of the ticks. Integrative taxonomy is the best for tick identification.