Identification of Shewanella baltica as the Most Important H 2 S-Producing Species during Iced Storage of Danish Marine Fish

Abstract

ABSTRACT Shewanella putrefaciens has been considered the main spoilage bacteria of low-temperature stored marine seafood. However, psychrotropic Shewanella have been reclassified during recent years, and the purpose of the present study was to determine whether any of the new Shewanella species are important in fish spoilage. More than 500 H 2 S-producing strains were isolated from iced stored marine fish (cod, plaice, and flounder) caught in the Baltic Sea during winter or summer time. All strains were identified as Shewanella species by phenotypic tests. Different Shewanella species were present on newly caught fish. During the warm summer months the mesophilic human pathogenic S. algae dominated the H 2 S-producing bacterial population. After iced storage, a shift in the Shewanella species was found, and most of the H 2 S-producing strains were identified as S. baltica . The 16S rRNA gene sequence analysis confirmed the identification of these two major groups. Several isolates could only be identified to the genus Shewanella level and were separated into two subgroups with low (44%) and high (47%) G+C mol%. The low G+C% group was isolated during winter months, whereas the high G+C% group was isolated on fish caught during summer and only during the first few days of iced storage. Phenotypically, these strains were different from the type strains of S. putrefaciens , S. oneidensis , S. colwelliana , and S. affinis , but the high G+C% group clustered close to S. colwelliana by 16S rRNA gene sequence comparison. The low G+C% group may constitute a new species. S. baltica , and the low G+C% group of Shewanella spp. strains grew well in cod juice at 0°C, but three high G+C Shewanella spp. were unable to grow at 0°C. In conclusion, the spoilage reactions of iced Danish marine fish remain unchanged (i.e., trimethylamine-N-oxide reduction and H 2 S production); however, the main H 2 S-producing organism was identified as S. baltica .