| Summary: | Deep spoilage is a cyclical and costly problem for the meat industry. Mianning ham is a famous dry-cured meat product in Sichuan, China. The aim of this work was to investigate the physicochemical characteristics, sources of odor, and associated microorganisms that cause spoilage of Mianning ham. High-throughput sequencing and solid-phase microextraction–gas-chromatography (SPME-GC-MS) techniques were used to characterize the physicochemical properties, microbial community structure, and volatile compounds of spoiled Mianning ham and to compare it with normal Mianning ham. The results showed that spoiled ham typically had higher moisture content, water activity (a<sub>w</sub>), and pH, and lower salt content. The dominant bacterial phylum detected in deeply spoiled ham was Firmicutes (95.4%). The dominant bacterial genus was <i>Clostridium_sensu_stricto_2</i> (92.01%), the dominant fungal phylum was Ascomycota (98.48%), and the dominant fungal genus was <i>Aspergillus</i> (84.27%). A total of 57 volatile flavor substances were detected in deeply spoiled ham, including 11 aldehydes, 2 ketones, 6 alcohols, 10 esters, 20 hydrocarbons, 6 acids, and 2 other compounds. Hexanal (279.607 ± 127.265 μg/kg) was the most abundant in deeply spoiled ham, followed by Butanoic acid (266.885 ± 55.439 μg/kg) and Nonanal (165.079 ± 63.923 μg/kg). <i>Clostridium_sensu_stricto_2</i> promoted the formation of five main flavor compounds, Heptanal, (E)-2-Octenal, 2-Nonanone, Hexanal, and Nonanal, in deeply spoiled ham by correlation analysis of microbial and volatile flavor substances.
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