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The present study was conducted to assess the seroprevalence of toxoplasmosis in small ruminants in Cholistan desert, Rahim Yar Khan and Rajan Pur districts of Southern Punjab, Pakistan to compare the rates of infections at species level and also to evaluate the relationship of T. gondii infection with age, gender and flock size of animals. The blood samples were collected randomly from 1200 small ruminants and examined for the prevalence of Toxoplasma gondii infection by using latex agglutination test. Out of total of 1200 blood samples collected from small ruminants (31.41%) were found positive for anti-T. gondii antibodies. The infection were found higher in sheep (37.31%) than in goats (29.13%), whereas the infection were almost the same in male and female small ruminants. A positive correlation was found in the age and flock size of small ruminants and prevalence of toxoplasmosis.