Shewanella　putrefaciens　is　a　typical　spoilage　bacteria　organism　in　seafood.　The　adhesion　ability　of　three　S.　putrefaciens　strains　(HR-15,　JR-18,　HC-71)　isolated　from　putrefied　tilapia　were　evaluated　by　mucus　adhesion　in　vitro　and　intestinal　adhesion　in　vivo.　The　results　of　the　spoilage　of　the　inoculated　fish　fillets　and　spoilage　of　the　refrigerated　fish　both　showed　that　the　adhesion　ability　of　S.　putrefaciens　was　positively　correlated　with　the　spoilage　ability.　High-throughput　sequencing　and　GC-MS　results　showed　that　S.　putrefaciens　with　high　adhesion　ability　also　significantly　changed　the　intestinal　flora　of　fish,　causing　an　increase　in　the　intestinal　bacteria　such　as　Plesionomas,　Macellibacteroides,　Acinetobacter,　and　Legionella,　which　then　led　to　the　increase　in　volatile　substances　such　as　low-grade　aldehydes,　alcohols,　and　ketones　in　the　fish,　serious　fatty　acid　oxidation,　and　excitement　of　the　fishy　smell.　©　2022　by　the　authors.　Licensee　MDPI,　Basel,　Switzerland.