EFFECT OF DIFFERENT CONCENTRATION OF TREHALOSE ON DOG SPERMATOZOA MORPHOLOGICAL CHANGES

Abstract

The aim of our work was to evaluate the large dog breeds ejaculate diluted with addition of trehalose with a focus on morphological changes. We used following breeds: German boxer, Greyhound, Leonberger, Border Collie. In our experiment different concentration of trehalose and cultivation temperature were used: 20 mg.ml-1 – TA; 5 mg.ml-1 – TB; 1.25 mg.ml-1 – TC; 0.312 mg.ml-1 – TD; control group – TK at 5°C and 20 mg.ml-1 – TAT; 5 mg.ml-1 – TBT; 1.25 mg.ml-1 – TCT; 0.312 mg.ml-1 – TDT; control group – TKT at 37°C. The occurrence of morphologically abnormal spermatozoa was quantified microscopically at 1000x magnification, and the following abnormal morphological changes were evaluated: SH – separated heads, AC – acrosomal changes, KTF – knob-twisted flagellum, FT – flagellum torso, FB – flagellum ball, LH – large head, SH – small head, PPS – % of physiological spermatozoa and PMC – % of morphological changed spermatozoa.
A higher number of normal spermatozoa (PPS) was found in the control group (TK) (84.33%) and the lowest number was detected in the TB group (79.92%). Morphological analysis showed similar occurrence of morphological abnormalities in all groups at 5°C with no significant differences (p>0.05).
The highest percentage of normal spermatozoa was found in the control group (TKT) (87.75%). The percentage of abnormal (morphologically changed – PMC) spermatozoa was the highest in the TAT group (18.33%). Significant differences were observed only in one parameter – acrosomal changes, between groups: TAT (5.41%) and TBT (3.25%) (p<0.05); TAT (5.41%) and TCT (2.83%) (p<0.01); group TAT (5.41%) compared to TDT (2.16%) and control group (TKT; 2.16%) (p<0.001). In other observed parameters no significant differences were found, but percentage of other parameters correlated with increasing concentration of trehalose.
Results of our study showed that morphological changes of spermatozoa were elevated in case of higher trehalose. Only a few of our results were statistically significant, therefore further research is needed.