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ABSTRACT The objective of this research was to determine the influence of heat stress, using the temperature-humidity index (THI), on the production and quality of native sperm of bulls. The effect of heat stress on the quantity of semen (mL), density of ejaculate (number of spermatozoa, 106/mL), gross sperm motility (1-5), number of frozen doses, and motility after freezing was analysed in 1,017 sperm samples taken from 32 Holstein-Friesian bulls, in the 2017-2019 period, at the Centre for Reproduction and Embryo Transfer in Serbia. The lowest amount of ejaculate (4.18±1.95 mL) and the lowest density of ejaculate (1,189.19±668.23 × 106/mL) were recorded under conditions of very strong heat stress on the day of semen collection. The level of heat stress measured on the day of semen collection did not affect sperm gross motility, number of frozen doses, and motility after freezing. The level of heat stress at the beginning of spermatogenesis, measured 60 days before semen collection, did not affect the amount of ejaculate and motility of spermatozoa after freezing, but at very strong stress, the lowest density of ejaculate (1,170.34±680.27 × 106/mL) and gross motility of spermatozoa were found (2.91±0.96). The lowest number of doses per ejaculate was recorded in conditions of moderate heat stress (396.6±157.71). Bulls older than 36 months had the best results according to all tested parameters of native sperm production and quality. The year in which the bulls produced semen did not affect density of ejaculate and sperm motility. The season of semen collection did not significantly affect the production and quality of native sperm, due to the practice of exploiting only bulls with the best sperm quality during the summer. temperaturehumidity temperature humidity THI, THI , (THI) mL, mL (mL) 106/mL, 106mL 106/mL 106 15, 15 1 5 (1-5) 1017 017 1,01 3 HolsteinFriesian Holstein Friesian 20172019 2017 2019 2017-201 period Serbia 4.18±1.95 418195 4 18 95 (4.18±1.9 1,189.19±668.23 11891966823 189 19 668 23 (1,189.19±668.2 spermatogenesis 6 1,170.34±680.27 11703468027 170 34 680 27 (1,170.34±680.2 2.91±0.96. 291096 2.91±0.96 . 2 91 0 96 (2.91±0.96) 396.6±157.71. 396615771 396.6±157.71 396 157 71 (396.6±157.71) summer (THI (mL 10 (1-5 101 01 1,0 2017201 201 2017-20 4.18±1.9 41819 9 (4.18±1. 1,189.19±668.2 1189196682 66 (1,189.19±668. 1,170.34±680.2 1170346802 17 68 (1,170.34±680. 29109 2.91±0.9 (2.91±0.96 39661577 396.6±157.7 39 7 (396.6±157.71 (1- 1, 201720 20 2017-2 4.18±1. 4181 (4.18±1 1,189.19±668. 118919668 (1,189.19±668 1,170.34±680. 117034680 (1,170.34±680 2910 2.91±0. (2.91±0.9 3966157 396.6±157. (396.6±157.7 (1 20172 2017- 4.18±1 418 (4.18± 1,189.19±668 11891966 (1,189.19±66 1,170.34±680 11703468 (1,170.34±68 291 2.91±0 (2.91±0. 396615 396.6±157 (396.6±157. ( 4.18± 41 (4.18 1,189.19±66 1189196 (1,189.19±6 1,170.34±68 1170346 (1,170.34±6 29 2.91± (2.91±0 39661 396.6±15 (396.6±157 4.18 (4.1 1,189.19±6 118919 (1,189.19± 1,170.34±6 117034 (1,170.34± 2.91 (2.91± 3966 396.6±1 (396.6±15 4.1 (4. 1,189.19± 11891 (1,189.19 1,170.34± 11703 (1,170.34 2.9 (2.91 396.6± (396.6±1 4. (4 1,189.19 1189 (1,189.1 1,170.34 1170 (1,170.3 2. (2.9 396.6 (396.6± 1,189.1 118 (1,189. 1,170.3 117 (1,170. (2. 396. (396.6 1,189. 11 (1,189 1,170. (1,170 (2 (396. 1,189 (1,18 1,170 (1,17 (396 1,18 (1,1 1,17 (39 1,1 (1, (3