# Potential Prebiotic and Anti-Obesity Effects of _Codium fragile_ Extract
## 3 Results
### Effect of Fermentation on Cell Viability Count and Acidity of CFE  
CFE fermentation was performed, and cell viability count and acidity observed over time. The changes to cell viability during fermentation are shown in Figure 1. The viable cell counts of _Lp_ in CFE supplemented media were 9.26 and 7.39 log CFU/mL at 24 and 48 h, respectively. For _Lh_, the viability counts in CFE supplemented medium were 9.41 and 7.81 log CFU/mL at 24 and 48 h, respectively. For fermentation with glucose, the viability counts for _Lp_ and _Lh_ were 9.35 and 9.41 log CFU/mL at 24 h and were 8.06 and 8.89 log CFU/mL at 48 h, respectively. The changes in acidity of the CFE fermentation solution are shown in Figure 2. For both _Lp_ and _Lh_, in CFE supplemented media, fermentation led to gradual increases in acidity. For these lactic acid bacteria, growth results in lactic acid production, which explains the increase in acidity from 0.2 % to 2.0 % after 48 h fermentation. For _Lp_, the \begin{table}
\begin{tabular}{c c c c c} \hline \hline  & & \multicolumn{3}{c}{**Absorbance of Culture (OD600)**} \\ \cline{3-5}
**Strains** & & **Glucose** & **GOS** & **CFE** \\ \hline \multirow{5}{*}{Probiotics} & _Bifidobacterium bifidum_ KTCT 3202 & 1.45 \(\pm\) 0.05 8* & 1.60 \(\pm\) 0.01 h & 1.53 \(\pm\) 0.01 h \\  & _Bifidobacterium animalis_ subsp. _lactis_ Bb-12 & 1.26 \(\pm\) 0.07 e & 1.36 \(\pm\) 0.07 f & 1.16 \(\pm\) 0.05 d \\  & _Lactobacillus rhamns_ GG ATCC 53103 & 1.26 \(\pm\) 0.05 e & 1.38 \(\pm\) 0.03 f & 1.19 \(\pm\) 0.01 d \\  & _Lactobacillus helictivacs_ ATCC 15807 & 1.18 \(\pm\) 0.01 d & 1.26 \(\pm\) 0.03 e & 1.33 \(\pm\) 0.01 f \\  & _Lactobacillus plantarum ATCC 14917_ & 1.26 \(\pm\) 0.01 e & 1.15 \(\pm\) 0.01 d & 1.34 \(\pm\) 0.01 f \\  & _Lactobacillus brevis_ KTCT 3498 & 1.41 \(\pm\) 0.01 s & 1.47 \(\pm\) 0.01 s & 1.45 \(\pm\) 0.01 s \\ \hline \multirow{2}{*}{Pathogens} & _Bacillus cereus_ ATCC 14579 & 1.04 \(\pm\) 0.04 c & 0.85 \(\pm\) 0.07 b & 0.72 \(\pm\)0.05 a \\  & _Escherichia coli_ ATCC 25922 & 0.98 \(\pm\) 0.04 c & 0.92 \(\pm\) 0.03 c & 0.65 \(\pm\) 0.06 a \\  & _Salmonella Typhii_ ATCC 19430 & 1.05 \(\pm\) 0.06 c & 0.77 \(\pm\) 0.06 b & 0.59 \(\pm\) 0.05 a \\ \hline \multicolumn{5}{c}{Values are mean \(\pm\) SD of three independent experiments. Sample means with different lower-case letters for the same index are significantly different at \(p\) \(<\) 0.05.} \\ \end{tabular}
\end{table} 
Table 2: Growth (OD600) of probiotics and food-born pathogenic bacteria grown in brain heart infusion (BHI) medium supplemented with CFE. Figure 1: Changes in cell viability counts of _Lp_ (**a**) and _Lh_ (**b**) with glucose or CFE. Values are expressed as mean \(\pm\) SD. acidity level was similar between the CFE and glucose-supplemented media, with an increase from 0.2 % to 2.0% over the course of fermentation. For _Lh_, the acidity level was lower after fermentation in CFE-supplemented medium when compared with that of glucose-supplemented medium. Table 3 shows SCFAs content after in vitro fermentation of CFE by mouse feces. When CFE was employed as the carbon source, acetic acid concentration was significantly higher than that of the mouse fecal-fermented sample. Butyric acid and propionic acid were not detected. Table 4 shows TPC, TFC, and DPPH activity in glucose or CFE supplemented media before and after fermentation by probiotics. Fermentation cultures grown in CFE-supplemented medium showed higher TPC, TFC, and DPPH activity compared to cultures grown in glucose-supplemented medium. After fermentation, TPC, TFC, and DPPH activity had significantly increased compared to non-fermentation.