This paper aims to characterize and interpret the trends in reserves, resources, and mine production of diatomite in the Czech Republic in last two decades. With more than 2.4 million tonnes of total reserves, 1.6 million tonnes of exploitable (recoverable) reserves, and average annual production of 35 kt, diatomite is not one of the key industrial minerals of the Czech Republic, which ranks among the top 10 European producers. Historical diatomite deposits were situated within the Cheb Basin, where the Holocene Hájek diatomite deposit was abandoned in 1955 because of the establishment of the Soos National Natural Monument. The group of Tertiary diatomite deposits situated in the Central Bohemian Upland ceased extraction when the last deposit (Kučlín) was abandoned in 1966 after depletion of reserves. The last group of diatomite deposits is located within the Southern Bohemian basins, where the last productive deposit, Borovany-Ledenice, is situated. Miocene diatomites are extracted by open pit mining there. Production of crude diatomite varied from 0 to 83 kt, with an average of 35 kt, between 1999 and 2018 according to stockpiles. Raw diatomite is classified into two groups according to the chemical-technological properties. Better-quality diatomite (SiO2 ≥ 72%, Al2O3 ≤ 15%, Fe2O3 < 2.4%, bulk density 450 kg/m3, loss on ignition < 8%) is processed for filtration in the food industry (brewery, wine, and raw fruit juices). Material with lower quality is used in combination with bentonite to prepare cat litter products.