The objective of this study was to optimize interrupted slow-freezing protocols for African catfish semen. Semen diluted with methanol and extender was frozen in 1-ml vials in a programmable freezer. The temperatures of the freezer (Tchamber) and of the semen (Tsemen) were measured simultaneously. We first tested two-step freezing protocols with different cooling rates (−2, −5, and −10°C/min) and different temperatures at plunging into liquid N2. The difference between Tsemen and Tchamber increased with faster cooling rates. In all programs, survival of spermatozoa, expressed as hatching rates, increased from near zero when Tsemen at plunging was higher than −30°C to values equal to those of control when Tsemen at plunging was equal to or lower than −38°C. The inclusion of an isothermal holding period before plunging into liquid N2 (three-step freezing protocols) resulted in an equilibration between Tsemen and Tchamber and improved semen survival. Semen could be plunged at temperatures as high as −36°C when cooled at −5 or −10°C/min, without compromising postthaw semen survival. Cooling at −2°C/min in combination with a 5-min holding period reduced postthaw survival. We conclude that with slow cooling rates of −2 to −5°C/min, hatching rates can be maximized by plunging as soon as Tsemen reaches −38°C. The isothermal holding period is beneficial when faster rates are used. A simple and efficient protocol for freezing African catfish semen can be obtained by cooling at a rate of −5 to −10°C/min combined with a 5-min holding period in the freezer, at −40°C