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<p>Abstract</p> <p>Background</p> <p>Toxoplasmosis is a zoonotic parasitic disease caused by the protozoan <it>Toxoplasma gondii,</it> which infects almost all warm-blooded animals, including humans, with a worldwide distribution. However, little is known of <it>T. gondii</it> seroprevalence in pet dogs in Kunming, the capital of Yunnan Province, southwest China. The objective of this investigation was to estimate the seroprevalence of <it>T. gondii</it> infection in pet dogs in this area.</p> <p>Methods</p> <p>A total of 611 serum samples were collected from 7 pet hospitals in Kunming, and assayed for <it>T. gondii</it> antibodies by the indirect haemagglutination (IHA) using a commercially-marked kit.</p> <p>Results</p> <p>132 (21.6%) pet dogs were positive for <it>T. gondii</it> antibodies, and the seroprevalence ranged from 17.3% to 34.7% among different sampling regions, the difference was statistically significant (<it>P</it> < 0.05). The <it>T. gondii</it> seroprevalence in female and male dogs were 20.8% and 22.4%, respectively, the difference was not statistically significant (<it>P</it> > 0.05). The seroprevalence ranged from 17.5% to 23.6% among different age groups, but the difference was not statistically significant (<it>P</it> > 0.05), and there were no interactions in statistics (<it>P</it> > 0.05) between gender and age of pet dogs in the region.</p> <p>Conclusions</p> <p>The findings of the present survey indicate high <it>T. gondii</it> seroprevalance in pet dogs in Kunming, southwest China, posing significant public health concern. It is necessary to enhance integrated strategies and measures to prevent and control <it>T. gondii</it> infection in pet dogs in this area.</p>