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Objective:To investigate the infection ofFasciola gigantica(F. gigantica) in domestic cattle from ChiangMai province and molecular confirmation usingITS-2 region.Methods:The liver and gall bladder ofBubalus bubalis(B. bubalis) andBos taurus(B. taurus) from slaughterhouses were examined adult worms and prevalence investigation.The species confirmation with phylogenetic analysis usingITS-2 sequences was performed by maximum likelihood andUPGMA methods. Results:The total prevalences of infection inB. bubalis andBubalus taurus(B. taurus) were 67.27% and52.94% respectively.The respective prevalence in bothB. bubalis andB. taurus were acquired fromDoi-Saket,Muang, andSanpatong districts, with81.25%,62.50% and60.00% for B. bubalis and62.50%,50.00% and47.06% forBos taurus respectively.The species confirmation ofF. gigantica and some related species by basing on maximum likelihood andUPGMA methods used,4 groups of trematodes weregenerated, firstF. gigantica group including specimen ofChiang Mai, second2 samples of F. hepatica, third group of3 rumen flukes;Orthocoelium streptocoelium, F. elongatus andParamphistomum epliclitum and fourth group of3 minute intestinal flukes;Haplorchis taichui, Stellantchasmu falcatus,Haplorchoides sp. and liver fluke;Opisthorchis viverrini respectively.Conclusions:These results can be confirmed theGiant liver fluke which mainly caused fascioliasis inChiangMai was identified asF. gigantica and specimens were the same as those ofF. gigantica recorded in other different countries.Nucleotide sequence of ITS-2 region has been proven as effective diagnostic tool for the identification ofF. gigantica.