Milk production is a significant area of the Czech agrarian sector. A long-term sustainability of dairy cattle operations is difficult to accomplish without an adequate level of profitability, which is influenced by a number of factors. For the total of 106 dairy farms in 2017, the observed total cost was CZK 73 521 per cow per year, i.e. CZK 8.90 per liter of milk at the milk production of 8 262 liters sold per cow per year. The costs per cow tended to increase over the last period. In 2017, total costs were reduced by CZK 0.47 per liter of milk as a result of deducting the price of calves and manure sold. The milk price of CZK 8.64 per liter and government subsidies of CZK 0.84 per liter resulted in the profit of 1.05 CZK per liter, i.e. overall profitability of 12.4%. The analysis confirmed the dependence of economic indicators on marketed milk production and the size of herd. In addition, the level of profitability was also different according to the breed used and the type of production area. The break-even point without subsidies in 2017 for milk price was CZK 8.43 per liter and for marketed milk yield of 7,866 liters per cow per year, which is 5.0% lower than the actual average of dairy farms included in analysis due to the positive profitability of milk production