Male tigers (Panthera tigris altaica) in captivity copulate alternatively with an estrous female, suggesting a potential for heteropaternity as an effective reproductive strategy to maximize genetic diversity of offspring. We analyzed microsatellites to test and compare the genetic output of multiple male mating (simultaneous polyandry) and single male mating (monogamy) with a female in a captive population. Simultaneous polyandry resulted in heteropaternity in 66.7% observed litters. No significant differences between parental populations and between offspring populations were detected in the number of alleles (A), expected heterozygosity (H e ), number of effective alleles (N e ) per locus and standard individual heterozygosity (SH) (P>0.05 for all 4 indexes). Comparisons showed no significant reduction of A, H o , H e and SH from parental population to offspring population for the two mating modes (P>0.05) except for SH in polyandrous families (P=0.029). However, such reduction was equivalent to single mating families when the influence of relatedness was eliminated using effective SH (E SH ) (P>0.05). These results highlight an alternative strategy for managing captive populations of tiger and other wild felids in which animals are combined at one location allowing for copulation by multiple males to encourage heteropaternity in favor of maintained genetic diversity among offspring.