of ammonia was measured for each of the test conditions as shown in Figure 10 (individual data are shown in Table 5). 

Table 5 - Ammonia (mM) measured in TIM-2 samples after adding the test products sugar cane bagasse (3.5g and 7g) and cellulose for all TIM-2 runs 

The production of ammonia was highest for 3.5 g (125.3 – 1.7 mmol), followed by 7 g sugar cane bagasse (122.3 – 3.4 mmol) and cellulose (1 19.3 – 1.7 mmol). ANOVA indicated significantly different ammonia values between the treatment groups. A Tukey’s posthoc test indicated furthermore a significant difference between 3.5 g and cellulose after 32h of experiment, between 3.5 g and cellulose as well as between 7 g and 3.5 g after 56h and between 3.5 g and cellulose after 80h. 

## Microbiota composition 

To test the effect of the test products on the microbiota composition, 16S sequencing was performed in TIM-2 luminal samples after 80 h of experimental period. 

Bacteroides are the most abundant bacteria present from all test conditions in TIM-2 after 80h. Bacteroides are known for their capability of degrading complex carbohydrates. The changes in relative abundances of bacteria for sugar cane bagasse compared to cellulose were calculated as fold changes and presented in Figure 13 (see equation 1). 

A fold change above 2.0 indicates a higher relative abundance for the sugar cane compared to cellulose after 80h in TIM-2. On the other hand, a fold change below 0.5 indicates lower relative abundance for sugar cane compared to cellulose and fold changes between 0.5 and 2.0 indicate that there is no significant change. 

