The Canadian wood preservation industry is at a competitive disadvantage in both domestic and international markets because our major species (e.g. spruce and pine) are difficult to impregnate as they are characterized by a thin sapwood band and refractory heartwood which limits chemical penetration. Achieving adequate penetration may be even more challenging for the next generation of carbon-based and micronized preservatives. Biological incising with Dichomitus squalens, developed in Austria in the 1990s, increased permeability in European spruce. More recently, studies done on commercial sized pine and spruce with a Canadian isolate of D. squalens under non-sterile conditions showed through-treatment (19 mm penetration) with 1.7% ACQ after six weeks in spruce samples; however, strength loss was slightly higher in some samples than what is found with conventional incising. Penetration was more variable in pine. A shorter incubation time would be desirable in an industrial setting. The current study was undertaken in order to determine if the processing time could be shortened by optimizing incubation temperature. Three temperatures were tested on small pine blocks using a soil block test method. Samples were exposed for between two and six weeks. ACQ-D treatment was used in this experiment as a surrogate for carbon-based preservatives because copper is easier to detect than colourless carbon-based preservatives. Optimal temperature for incubation appeared to cover a wide range around 30 °C and no growth inhibition was encountered at 35 °C suggesting that temperature variability in an industrial setting should not inhibit a biological incising operation. Uptake and penetration was variable. After 5 weeks seventy percent of pine samples reached a minimum of 5 mm penetration, and 50 % of the samples reached a minimum of 10 mm penetration. Optimal incubation time still appears to be over six weeks in pine to reach minimum 10mm penetration requirements in over 80% of samples.