Microbial diversity and dominant bacteria causing spoilage during storage and processing of the Antarctic krill, Euphausia superba
Tóm tắt
The Antarctic region is known for its ecological conditions and the presence of some of the rarest microorganisms on earth. Antarctic krill rapidly degrades while at rest or during transport due to high-activity enzymes in its flesh or from microbes. In this study, we analyzed the microbial diversity of the Antarctic krill, Euphausia superba, and identified the dominant bacteria that cause spoilage during storage and transport using culture-independent high-throughput sequencing and microbial pure culture methods. Antarctic krill samples were incubated at temperatures of 0 °C, 4 °C, 16 °C, and 25 °C to determine the microbial diversity and abundance in decaying marine krill samples. Metagenome high-throughput sequencing indicated that, under incubation at 4 °C and 0 °C for 24 h, the microbial diversity of the Antarctic krill samples was high, with high species richness of microbes adapted to the low-temperature environment, as indicated by the overwhelming dominance of Enterococcus and Bacillus. On the other hand, Psychrobacter became the dominant bacteria at storage temperatures of 16 °C and 25 °C. Quantitative PCR results demonstrated that the microbial copy number increased as the temperature increased from 0 to 25 °C, and the quantity of bacteria was much greater than that of fungi under the same storage conditions. Microbial pure culture methods demonstrated that Psychrobacter was the dominant genus at each incubation temperature after 24 h of storage. Psychrobacter sp. and Psychrobacter-like strains isolated from the Antarctic krill samples exhibited protease activity. Our laboratory results indicated that Psychrobacter was the primary cause of spoilage of the Antarctic krill, E. superba. The results from ITS rDNA fungal sequencing showed that unclassified Saccharomycetes were the predominant microbes in all the Antarctic krill samples.
Tài liệu tham khảo
Anagnostopoulos GD (1979) Microbial life in extreme environments. Nature 279:658
Anheller JE, Hellgren L, Karlstam B, Vincent J (1989) Biochemical and biological profile of a new enzyme preparation from Antarctic krill (E. superba) suitable for debridement of ulcerative lesions. Arch Dermatol Res 281:105–110
Auerswald L, Meyer B, Teschke M, Hagen W, Kawaguchi S (2015) Physiological response of adult Antarctic krill, Euphausia superba, to long-term starvation. Polar Biol 38:763–780
Benjamin DC, Kristjánsdóttir S, Gudmundsdóttir A (2001) Increasing the thermal stability of euphauserase A cold-active and multifunctional serine protease from Antarctic krill. Eur J Biochem 1:127–131
Bucht A, Karlstam B (1991) Isolation and immunological characterization of three highly purified serine proteinases from Antarctic krill (Euphausia superba). Polar Biol 11:495–500
Caporaso JG, Kuczynski J, Stombaugh J, Bittinger K, Bushman FD, Costello EK, Fierer N, Peña AG, Goodrich JK, Gordon JI, Huttley GA, Kelley ST, Knights D, Koenig JE, Ley RE, Lozupone CA, McDonald D, Muegge BD, Pirrung M, Reeder J, Sevinsky JR, Turnbaugh PJ, Walters WA, Widmann J, Yatsunenko T, Zaneveld J, Knight R (2010) QIIME allows analysis of high-throughput community sequencing data. Nat Methods 5:335–336
Cappuccino JG, Sherman N (1983) Microbiology: a laboratory manual. Addison Wesley, Canada
Cui XQ, Zhu GL, Liu HS, Jiang GL, Wang Y, Zhu WM (2016) Diversity and function of the Antarctic krill microorganisms from Euphausia superba. Sci Rep 6:36496
Denner EB, Mark B, Busse HJ, Turkiewicz M, Lubitz W (2001) Psychrobacter proteolyticus sp. nov., a psychrotrophic, halotolerant bacterium isolated from the Antarctic krill Euphausia superba Dana, excreting a cold-adapted metalloprotease. Syst Appl Microbiol 24:44–53
Donachi SP, Zdanowski MK (1998) Potential digestive function of bacteria in krill Euphausia superba stomach. Aquat Microb Ecol 14:129–136
Ellingsen TE, Mohr V (1987) Biochemistry of the autolytic processes in Antarctic krill post mortem. Biochem J 246:295–305
Fevolden SE (2010) Krill: biology, ecology and fisheries (Fish and aquatic resources series 6). Fish Fish 3:57–57
Friedmann EI, Thistle AB (1993) Antarctic microbiology. Wiley-Liss, New York
Ikeda T, Dixon P (1982) Body shrinkage as a possible over-wintering mechanism of the Antarctic krill, Euphausia superba Dana. J Exp Mar Biol Ecol 62:143–151
Kawamura Y, Nishimura K, Matoba T, Yonezawa D (1984) Effects of protease inhibitors on the autolysis and protease activities of Antarctic krill. Agric Biol Chem 48:923–930
Kraft AA (1992) Psychrotrophic bacteria in foods: disease and spoilage. CRC, Boca Raton
Li L, Li ZQ, Li HQ, Bi SS, Xu JJ, Li B, Zhou TY, Lu W (2011) Absolute quantification of induced mRNA expression of CYP3A1 and CYP3A2 in rat liver using quantitative real time PCR assay. J Chin Pharm Sci 6:597–603
Liu JT, Lu XL, Liu XY, Gao Y, Hu B, Jiao BH, Zheng H (2013) Bioactive natural products from the Antarctic and Arctic organisms. Mini Rev Med Chem 13:617–626
Logares R, Sunagawa S, Salazar G, Cornejo-Castillo FM, Ferrera I, Sarmento H, Hingamp P, Ogata H, Vargas C, Lima-Mendez G, Raes J, Poulain J, Jaillon O, Wincker P, Kandels-Lewis S, Karsenti E, Bork P, Acinas SG (2014) Metagenomic 16S rDNA Illumina tags are a powerful alternative to amplicon sequencing to explore diversity and structure of microbial communities. Environ Microbiol 16:2659–2671
Meyer B (2012) The overwintering of Antarctic krill, Euphausia superba, from an ecophysiological perspective. Polar Biol 35:15–37
Nadeem F, Oves M, Qari HA, Ismail IMI (2015) Red sea microbial diversity for antimicrobial and anticancer agents. J Mol Biomark Diagn 7:267–271
Nicol S, Foster J, Kawaguchi S (2012) The fishery for Antarctic krill recent developments. Fish Fish 13:30–40
Nishimura K, Kawamura Y, Matoba T, Yonezawa D (1983) Deterioration of Antarctic krill muscle during freeze storage. Agric Biol Chem 47:2881–2888
Rakusa-Suszczewski S, Zdanowski MK (1989) Bacteria in krill (Euphausia superba Dana) stomach. Acta Protozool 28:87–90
Rao S, Chan Y, Bugler-lacap DC, Bhatnagar A, Bhatnagar M, Pointing SB (2016) Microbial diversity in soil, sand dune and rock substrates of the Thar Monsoon Desert, India. Indian J Microbiol 56:1–11
Rastogi G, Sani RK (2011) Molecular techniques to assess microbial community structure, function, and dynamics in the environment. Microbes Microbial Technol 2:29–57
Rasuk MC, Fernandez AB, Kurth D, Contreras M, Novoa F, Poire D, Farias ME (2016) Bacterial diversity in microbial mats and sediments from the Atacama Desert. Microb Ecol 71:44–56
Siegel V (2000) Krill (Euphausiacea) life history and aspects of population dynamics. Can J Fish Aquat Sci 57:130–150
Siegel V (2010) The Antarctic krill: resource and climate indicator, 35 years of German krill research. J Appl Ichthyol 26:41–46
Sjödahl J, Emmer A, Karlstam B, Vincent J, Roeraade J (1998) Separation of proteolytic enzymes originating from Antarctic krill (Euphausia superba) by capillary electrophoresis. J Chromatogr B 705:231–241
Sjödahl J, Emmer A, Vincent J, Roeraade J (2002) Characterization of proteinases from Antarctic krill (Euphausia superba). Protein Expr Purif 26:153–161
Tou J, Jaczynski J, Chen YC (2007) Krill for human consumption: nutritional value and potential health benefits. Nutr Rev 65:63–77
Vick-Majors T, Achberger A, Santibanez P, Dore JE, Hodson T, Michaud AB, Christner BC, Mikucki J, Skidmore ML, Powell R (2016) Biogeochemistry and microbial diversity in the marine cavity beneath the McMurdo Ice Shelf, Antarctica. Limnol Oceanogr 61:572–586
Wang YC, Wang SS, Wang JF, Xue CH, Chang YG, Xue Y (2015) Preparation and anti-osteoporotic activities in vivo of phosphorylated peptides from Antarctic krill (Euphausia superba). Peptides 68:239–245
Wu ZQ, Wang JR, Shang XM, Yang ZQ, Jiang GL (2014) Purification and characterization of cold adapted trypsins from Antarctic Krill (Euphausia superba). Int J Pept Res Ther 20:531–543
Xia GH, Zhao Y, Yu Z, Tian Y, Wang Y, Wang S, Wang J, Xue C (2015) Phosphorylated peptides from Antarctic krill (Euphausia superba) prevent estrogen deficiency induced osteoporosis by inhibiting bone resorption in ovariectomized rats. Agric Food Chem 63:9550–9557
Zhang XY, Bao J, Wang GH, He F, Xu XY, Qi SH (2012) Diversity and antimicrobial activity of culturable fungi isolated from six species of the South China sea gorgonians. Microb Ecol 64:617–627