Objective To compare the impact of two anticoagulants (EDTA-3K and heparin lithium) on blood physiological parameters in miniature pigs. Methods Blood samples from 22 adult miniature pigs were collected and tested for physiological values using a MEK-7222K hematology analyzer. Differences in the values determined by using the two anticoagulants were analyzed. Results There were significant differences in values of some blood physiological parameters determined by the two anticoagulant groups, such as MON,MCV,MCHC,PLT,PCT,MPV,LYM,EOS,BAS,LYM %,MON%,EOS% and BAS%. There were significant differences in the latter 12 indexes (P<0.01) except for MON (P<0.05). Meanwhile, there were sex differences in values of 7 blood physiological indexes determined by EDTA-3K: PLT (P<0.05), NEUT (P<0.05),NEUT% (P<0.05),MON (P<0.01),EOS (P<0.01),MON% (P<0.01) and EOS% (P<0.01). There were significant differences in 8 indexes determined by heparin lithium: PDW (P<0.05), NEUT (P<0.05), LYM% (P<0.05), MON (P<0.01), EOS (P<0.01), MON% (P<0.01), NEUT% (P<0.01) and EOS% (P<0.01). Conclusion It is essential to choose proper anticoagulant before starting relevant tests, because there are significant differences in leukocyte and platelet counts in miniature pigs, determined with different anticoagulants.