# Enhanced Enzymatic Degradability of Livestock Blood Pretreated with Ultrasonic Technique
## 3 Results and Discussion
### Enzymatic Degradation of Untreated Livestock Blood
\begin{table}
\begin{tabular}{l c c c c c c} \hline \hline \multicolumn{2}{c}{**Parameters**} & **Case 1 \({}^{\text{c}}\)** & **Case 2 \({}^{\text{c}}\)** & **Case 3 \({}^{\text{c}}\)** & **Case 4 \({}^{\text{c}}\)** & **Case 5 \({}^{\text{c}}\)** \\ \hline \multirow{2}{*}{Protein (mg/mL)} & Raw \({}^{\text{a}}\) & 83.3 & 61.3 & 263.0 & 104.9 & 130.4 \\  & Ultrasonic-treated \({}^{\text{b}}\) & 12.5 & 13.8 & 78.4 & 27.8 & 33.8 \\ \hline \multirow{2}{*}{Amino acid (mg/mL)} & Raw \({}^{\text{a}}\) & 6.3 & 8.1 & 4.6 & 13.1 & 13.7 \\  & Ultrasonic-treated \({}^{\text{b}}\) & 15.1 & 13.4 & 56.9 & 54.6 & 47.5 \\ \hline \multirow{2}{*}{Protein Reduction Rate (\%) \({}^{\text{d}}\)} & Raw \({}^{\text{a}}\) & 78.5 & 84.2 & 32.1 & 72.9 & 66.3 \\  & Ultrasonic-treated \({}^{\text{b}}\) & 96.1 & 95.7 & 75.6 & 91.3 & 89.5 \\ \hline \multirow{2}{*}{Amino acid  Conversion Rate (\%) \({}^{\text{e}}\)} & Raw \({}^{\text{a}}\) & 1.6 & 2.1 & 1.19 & 3.34 & 3.5 \\  & Ultrasonic-treated \({}^{\text{b}}\) & 4.7 & 4.2 & 17.7 & 17.0 & 14.8 \\ \hline \multirow{6}{*}{
\begin{tabular}{} \end{tabular} } & \({}^{\text{a}}\) Livestock blood after grinding treatment (grinding speed: 10,000 rpm; grinding duration: 10 min); \({}^{\text{b}}\) Livestock blood after grinding and ultrasonic treatment (sample: 1 L; ultrasonic frequency: 20 kHz; ultrasonic density: 0.5 W/mL; ultrasonic duration: 30 mirr agitation: 200 rpm); \({}^{\text{c}}\) treatment conditions: sample: 100 mL; temperature: 55 \({}^{\text{c}}\)C; reaction time: 4 h; \({}^{\text{d}}\) ratio of reduced protein compared to supplied protein concentration: (initial protein concentration – final protein concentration)/initial protein concentration) \(\times\) 100; \({}^{\text{e}}\) amino acid conversion rate compared to supplied protein concentration: (initial amino acid concentration – final amino acid concentration)/initial protein concentration) \(\times\) 100. \\ \hline \end{tabular}
\end{table} 
Table 4: Enzymatic degradability of untreated and ultrasonic-treated Livestock blood. cost-saving effect of the combined use of enzymes, it is considered most desirable to use a combination of both enzyme types for this process. The comparison of the results before and after ultrasound treatment revealed that the untreated blood samples exhibited a maximum amino acid conversion rate of only 13.7 mg/mL in all enzymatic reactions, while that of the ultrasound-treated blood samples increased to 56.9 mg/mL, which is more than four times higher than the amino acid conversion rate of the untreated blood. The comparison of the concentration changes over reaction time revealed that the ultrasound-treated samples had significantly higher protein reduction and amino acid conversion rates than the untreated samples. This finding sug  
Figure 4: Change in enzymatic degradability of raw and ultrasonic-treated Livestock blood according to reaction time; (**a**) Case 1: 2% Savinase®; (**b**) case 2: 2% Alcalase®; (**c**) Case 3: 2% Flavourzyme®; (**d**) Case 4: 1% Savinase® + 1% Flavourzyme®; (**e**) Case 5: 1% Alcalase® + 1% Flavourzyme® (untreated sample conditions—grinding speed: 10,000 rpm; grinding duration: 10 min; treated sample conditions—sample: 1 L; ultrasonic frequency: 20 kHz; ultrasonic density: 0.5 W/mL; ultrasonic duration: 30 min; agitation: 200 rpm). gests that ultrasonic treatment induces hemolysis in RBCs, facilitating enzymatic reactions and altering the protein structure, thus accelerating the enzyme hydrolysis reaction rate. This result is consistent with a report of [PERSON] et al. [20], who investigated a study on ultrasonic pretreatment process before the enzymatic hydrolysis of caprine milk samples, and reported that the concentration of water-soluble proteins in goat milk increased through ultrasonic pretreatment. They reported that ultrasonication enhanced the production of protein hydrolysates, which may be due to the evolution of complex cellular structures into simpler soluble matrices due to the cavitation effect of sonication.