Linguatulosis is one of the neglected parasitic zoonoses. Domestic ruminants are the important source for human infection. The main goal of the present investigation was to evaluate the prevalence of nymphal stages of L. serrata in domestic ruminant in Hamedan province, western Iran. The mesenteric and mediastinal lymph node of 1080 animals were taken randomly from different slaughterhouses of Hamedan province. All lymph nodes were examined for nymphal stage of L. serrata using parasitological methods (digestion technique). The nymph of L. serrata was found in 31.4% (69/220) of goats, 15% (60/400) of sheep, and 7.4% (34/460) of cattle (P=0.457). The infection rate in beef cattle, dairy cattle and native cattle was 1.3%, 6.4% and 12.6%, respectively (P=0.981). No evidence of correlation between gender, age groups, and L. serrata infection rate in sheep and cattle (P>0.05). In goats, the infection rate in >2yr old (48%) was higher than ≤2yr old (8.6%) (P<0.0001, OR=5.6); also the infection rate was 23.7% and 35.7% in male and female goats, respectively (P=0.065). This is the first report of linguatulosis in domestic animals from western Iran. The results indicate that linguatulosis may partly be responsible for economic losses in domestic ruminant husbandry in this region. Therefore, designing control strategies for a better management in animals farm to reduce the risk of zoonotic outbreaks is highly recommended.