# Enhanced Enzymatic Degradability of Livestock Blood Pretreated with Ultrasonic Technique
## 3 Results and Discussion  
### Protein Solubilization by Ultrasonic Pretreatment  
The results of ultrasonic treatment for the solubilization of proteins in Livestock blood are shown in Figure 2. At the irradiation densities of 0.1 and 0.3 W/mL, the maximum solubilization rate was approximately 91-93%. At the irradiation density of \(\geq\)0.5 W/mL, a maximum solubilization rate of 96-97% was achieved, although the density-dependent solubilization effect did not vary significantly. While higher ultrasonic irradiation densities are generally known to yield better ultrasound efficiency [32], our experiments did not yield any substantial difference in ultrasound efficiency with varying densities. Instead, we noticed that the solubilization rate increased as the ultrasonic duration increased. This difference may be attributed to the specific characteristics of the samples or the ultrasonic treatment methodology. In particular, blood can be easily hemolyzed even with minimal physical force, indicating a Limited impact of ultrasonic density [33]. Our results revealed that an ultrasonic density of 0.5 W/mL and a duration of 30 min are the optimal ultrasonic treatment conditions for the solubilization of Livestock blood. Table 3 presents the changes in the characteristics of Livestock blood before and after ultrasonic treatment. The untreated samples were not subjected to any processing, while the ultrasonic-treated samples were subjected to ultrasonic treatment at the optimal density and duration conditions of 0.5 W/mL for 30 min. The parameters analyzed were pH, electrolytes, chemical components, plasma hemoglobin content, protein solubilization rate, particle size, and microorganisms. The analysis revealed that the hemoglobin concentration in the plasma of the grinding treated samples was 5.59%, while that of the ultrasonic-treated samples was 14.63%, which corresponds to a solubilization rate of 95.91%. This result indicates that during the ultrasonic treatment, most of the hemoglobin RBCs were hemolyzed into plasma, resulting in an increased protein content in the plasma. Therefore, blood proteins can be solubilized without additional chemical treatments, making it conducive to efficient enzymatic hydrolysis in future biological processes. The analysis of particle size distribution in the blood samples before and after ultrasound treatment revealed that the average particle size of the untreated samples was 131.16 \(\upmu\)m, while that of the post-treated samples was 44.32 \(\upmu\)m. This result indicates that both grinding and ultrasonic treatments contributed to the destruction of RBCs and breakdown of high molecular weight substances in the blood. This is further supported by the protein content in the untreated and post-treated blood samples, which were 387.0 g/L and 319.2 g/L, respectively. As mentioned earlier, this decrease in protein content is attributed to the disruption of protein chains caused by ultrasonic treatment. The protein  
Figure 2: Changes in solubilization rate with different ultrasonic density and ultrasonic time (sample \(-1\) L/batch, homogenization at 10,000 rpm for 10 min, ultrasonic frequency at 20 kHz, agitation at 200 rpm). content in the plasma of the post-treated samples was 2.5 times higher than that of the untreated samples. These results confirm that ultrasonic treatment disrupts RBCs, thereby enhancing the solubilization of proteins. The reduced particle size in the post-treated blood samples can be attributed to the decrease in protein size, which, in turn, enhances enzymatic degradability.