## 3 Results and Discussion
### NGS Analysis
\begin{table}
\begin{tabular}{l l l l l l l l l l l l l l l l l l l l l l} \hline \multirow{3}{*}{} & \multirow{3}{*}{**Muscle**} & \multirow{3}{*}{**Type**} & \multirow{3}{*}{**0 Day**} & \multicolumn{6}{c}{**3\({}^{\circ}\)C**} & \multicolumn{6}{c}{**0\({}^{\circ}\)C**} \\ \cline{3-14}  & & & & & & & & & & & & & & & & & & & & & & & & & & \\ \cline{3-14}  & & & & & & & & & & & & & & & & & & & & & & & & & \\ \hline TVBN & OM & 15.5 & \(\uparrow\) & 16.2 & 16.2 & 17.7 & 20.3 & * & 16.2 & 17.1 & 17.1 & 17.1 & 16.3 & 17.1 & 16.9 & 17.2 & 19.7 & * &  
\begin{table}
\begin{tabular}{c c c c c c c c c c c c c c c c c c} \hline \hline  & \multirow{2}{*}{**Muscle**} & \multicolumn{6}{c}{**3 \({}^{\circ}\)C**} & \multicolumn{6}{c}{**0 \({}^{\circ}\)C**} & \multicolumn{6}{c}{**-3 \({}^{\circ}\)C**} \\ \cline{3-14}  & **Type** & \multicolumn{3}{c}{**0 Day**} & \multicolumn{3}{c}{**Storage Periods (Days)**} & \multicolumn{3}{c}{**Storage Periods (Days)**} & \multicolumn{3}{c}{**Storage Periods (Days)**} \\ \cline{3-14}  & & & & **3** & **6** & **10** & **14** & **3** & **7** & **14** & **21** & **7** & **17** & **27** & **37** & **50** \\ \hline \multirow{2}{*}{Observed OTUs} & OM & 117.0 & 73.0 \({}^{\circ}\) & 66.3 \({}^{\circ}\) & 26.3 \({}^{\circ}\) & 25.0 \({}^{\circ}\) & 65.3 \({}^{\circ}\) & 42.7 \({}^{\circ}\) & 28.7 \({}^{\circ}\) & 23.0 \({}^{\circ}\) & 60.7 \({}^{\circ}\) & 76.0 \({}^{\circ}\) & 46.0 \({}^{\circ}\) & 27.3 \({}^{\circ}\) & 21.0 \({}^{\circ}\) \\  & DM & 121.0 & 74.7 & 60.7 \({}^{\circ}\) & 26.3 \({}^{\circ}\) & 22.0 \({}^{\circ}\) & 80.7 & 34.0 \({}^{\circ}\) & 37.3 \({}^{\circ}\) & 21.7 \({}^{\circ}\) & 82.0 \({}^{\circ}\) & 69.3 \({}^{\circ}\) & 50.7 \({}^{\circ}\) & 33.3 \({}^{\circ}\) & 18.3 \({}^{\circ}\) \\ Shannon & OM & 54.3 & 52.5 & 4.66 & 35.4 \({}^{\circ}\) & 2.89 \({}^{\circ}\) & 52.7 & 37.5 \({}^{\circ}\) & 3.4 \({}^{\circ}\) & 3.19 \({}^{\circ}\) & 3.70 \({}^{\circ}\) & 4.73 \({}^{\circ}\) & 3.82 \({}^{\circ}\) & 3.21 \({}^{\circ}\) & 2.71 \({}^{\circ}\) \\  & DM & 51.5 & 47.9 & 4.11 & 3.29 \({}^{\circ}\) & 2.97 \({}^{\circ}\) & 5.34 \({}^{\circ}\) & 3.55 \({}^{\circ}\) & 34.1 \({}^{\circ}\) & 31.2 \({}^{\circ}\) & 4.94 \({}^{\circ}\) & 4.37 \({}^{\circ}\) & 3.41 \({}^{\circ}\) & 3.05 \({}^{\circ}\) & 25.3 \({}^{\circ}\) \\  & OM & 119.0 & 73.7 \({}^{\circ}\) & 67.0 \({}^{\circ}\) & 26.7 \({}^{\circ}\) & 25.0 \({}^{\circ}\) & 65.3 \({}^{\circ}\) & 43.3 \({}^{\circ}\) & 29.3 \({}^{\circ}\) & 23.3 \({}^{\circ}\) & 61.7 \({}^{\circ}\) & 76.7 \({}^{\circ}\) & 48.0 \({}^{\circ}\) & 29.0 \({}^{\circ}\) & 21.3 \({}^{\circ}\) \\  & DMFigure 1: Relative abundance (%) of bacterial genes in raw yellowish muscles during storage at different temperatures based on the classification of partial 1680 \(\mu\)TDNA sequences. OM, dorsal part of ordinary muscle; DM, dark muscle. Storage time is indicated by # day. The predominance of _Pseudomonas_ aligns with previous findings that associate this genus with the spoilage of refrigerated muscle products, such as crayfish and beef, owing to its ability to thrive at low temperatures and generate volatile metabolites responsible for sensory deterioration [45; 50]. [PERSON] et al. reported that _Psychrobacter_ and _Pseudomonas_ dominated the bacterial communities in red seabream after 12 days of ice storage [51], whereas [PERSON] et al. observed similar trends in thawed common cuttlefish stored at 2 \({}^{\circ}\)C in Greece [52]. [PERSON] et al. reported the dominance of _Pseudomonas_ in partially frozen European crucial carp after storage [53]. In the present study, _Pseudomonas_ emerged as the predominant spoilage bacterium across various temperatures and muscle types, which is consistent with previous findings. This hardy gram-negative bacterium thrives in various storage environments for aquatic products, leveraging its ability to metabolize lipids and proteins [54]. In addition to _Pseudomonas_, other bacteria identified after spoilage in this study included _Acinetobacter_, _Brochothrix_, and _Shewanella_.