The adhesion ability related to biofilm in Acinetobacter species is very pivotal for spoilage capacity in food processing and preservation. The biofilm on the surfaces of chicken-processing tables and the chicken itself increases the risk of microbial contamination and meat spoilage. This is primarily due to the robust resistance of Acinetobacter within biofilms to sanitizers and disinfectants. The association among adhesion and biofilm formation ability in Acinetobacter and chicken spoilage is of paramount importance for the poultry industry, which must prioritize product safety and quality by clarifying the connection among these three factors. Here, two strains of Acinetobacter, namely Acinetobacter seifertii 5D-3 and Acinetobacter haemolyticus F-2, isolated from broiler slaughter, were examined to analyze the heterogeneity of biofilm. The investigation focused on the adhesion and motility capacity of these strains to identify the underlying principle of biofilm formation, while the impact on spoilage capacity was assessed through series assays both in vitro and in situ. The results indicated that strain F-2 exhibited greater detrimental effects on chicken freshness, attributable to its enhanced adhesion and biofilm capabilities. Precisely, the total colony count for F-2 reached 5.2 log (CFU/g) by the fifth day, with an adhesion rate of 93 %. Furthermore, a comparative analysis of volatile compounds revealed that three specific substances-heptanal, methyl-heptenone, and octan-3-ol may differentially affect spoilage capacity between the two strains. This research unlocks valuable theoretical insights into the relationship between biofilm formation capacity and spoilage effects in Acinetobacter, offering practical implications for enterprise managers seeking to mitigate contamination by Acinetobacter and improve chicken freshness.