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The aim of this study was to identify the effect of ambient temperature and season of the year on conception difficulties in Holstein cattle. The experiment was conducted on the School farm in Žabčice. Data were collected from July 2016 to January 2017. Experimental population counted 263 breeding Holstein cows, which were inseminated during the observation period. Results of this study confirmed the effect of temperature and season on conception rates in dairy cows. In winter season, successful conception was recorded in 56.36 % of cows in average ambient temperature 1.48 °C, in summer season the conception rate was only 40.43 % in average daily temperature 18.96 °C. A more detailed analysis revealed the highest conception rates in the dairy cows in January (88.24 %) and February (91.67 %). On the contrary, the worst results were recorded in September (22.73 %) and October (28.57 %). Evaluation of conception rate according to ambient temperature intervals showed best results for the temperatures below 5 °C. A part of the study was also evaluation of an association between daily milk yield and conception rate. This hypothesis cannot be conclusively confirmed, because milk yield was mostly similar in both groups (pregnant and non-pregnant) on the days of insemination. The effect of lactation order and mean daily milk yield on the conception rate in cows was not confirmed. [ABSTRACT FROM AUTHOR]