The objective of this work was to study the effect of mixed culture solid substrate fermentation of C. tropicalis ZD-3 with A. niger ZD-8 on detoxification of cottonseed meal (CSM), and to investigate the effect of fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment on the reduction of free gossypol levels during mixed culture solid substrate fermentation of CSM. Experiment 1: Three groups of disinfected CSM substrate were incubated for 48 h after

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... noculation with either of the fungi C. tropicalis ZD-3, A. niger ZD-8 or mixed culture (C. tropicalis ZD-3 with A. niger ZD-8). One non-inoculated group was used as the control. Levels of initial and final free gossypol (FG), CP and in vitro CP digestibility were assayed. The results indicated that mixed culture fermentation was far more effective than single strain fermentation, which not only had higher detoxification rate, but also had higher CP content and in vitro digestibility. Experiment 2: CSM substrates were treated according to experimental variables including fermentation period, proportion of CSM in substrate, sodium carbonate, minerals and heat treatment, Then, the treated CSM substrates were inoculated with mixed culture (C. tropicalis ZD-3 with A. niger ZD-8) and incubated at 30°C for 36 h in a 95% relative humidity chamber. After fermentation ended, FG and CP content of fermented CSM substrate was assayed. The results showed that the appropriate fermentation period was 36 h, and the optimal proportion of CSM in substrate was 70%. Addition of sodium carbonate to CSM substrate was beneficial for fermentative detoxification. Heat treatment could facilitate fermentative detoxification, and supplementation with minerals was instrumental in reducing gossypol levels during mixed culture solid substrate fermentation of CSM. (