comparison of giemsa, leishman, and leishman-giemsa staining methods for evaluating fish blood smears

Background: the hematological panel provides valuable information about the physiologic status and health of fish in aquatic environments through optimal staining of the nucleus, cytoplasm, and cytoplasmic granules. objectives: our research focused on creating a new leishman-giemsa dye mixture specifically for fish blood smears and evaluating its effectiveness compared to standard staining techniques.methods: blood samples were taken from 20 healthy common carp to produce three groups of peripheral blood smears: one for leishman-giemsa dye and two for leishman’s and giemsa’s stains. two experienced clinical pathologists extensively examined all three types of blood smears based on four staining characteristics: nuclear characteristics of erythrocytes and leukocytes, cytoplasmic characteristics, and leukocyte granulation. the mean ratings from the two experts for each staining technique were compared. results: regarding the characteristics of rbc and wbc nuclei, the two conventional staining methods, leishman and giemsa, yielded substantially lower mean scores than the new leishman-giemsa staining method (p<0.05). leishman-giemsa staining enhanced the clarity of rbc and wbc nuclear characteristics. the new leishman-giemsa staining technique resulted in a statistically significant (p<0.05) difference in the cytoplasmic characteristics of fish wbc compared to the other two methods. conclusion: the results of this study demonstrated for the first time that fish blood cells stained with the novel leishman-giemsa method are more desirable. in addition, its nuclear and cytoplasmic staining is superior to that of conventional giemsa and leishman stains when used separately. the leishman-giemsa cocktail has a high index for air-dried smear discoloration.