[논문]A report of 12 unrecorded prokaryotic species isolated from gastrointestinal tracts and feces of various endangered animals in Korea

Kim, Pil Soo; Lee, Ki-Eun; Tak, Euon Jung; Kang, Myung-Suk; Bae, Jin-Woo

doi:10.12651/jsr.2020.9.1.035

A report of 12 unrecorded prokaryotic species isolated from gastrointestinal tracts and feces of various endangered animals in Korea 원문보기

Journal of species research, v.9 no.1, 2020년, pp.35 - 45

Kim, Pil Soo (Department of Biology and Department of Life and Nanopharmaceutical Sciences, Kyung Hee University) , Lee, Ki-Eun (Microorganism Resources Division, National Institute of Biological Resources) , Tak, Euon Jung (Department of Biology and Department of Life and Nanopharmaceutical Sciences, Kyung Hee University) , Kang, Myung-Suk (Biological Resources Utilization Department, National Institute of Biological Resources) , Bae, Jin-Woo (Department of Biology and Department of Life and Nanopharmaceutical Sciences, Kyung Hee University)

Abstract ▼ AI-Helper

In 2016 and 2017, as part of a comprehensive investigation to identify the prokaryotic species in Korea, a total of 12 bacterial strains were isolated from the gastrointestinal tract and/or fecal samples of four endangered species, including reptile, bird, and marine and terrestrial mammals. Phylogenetic analysis with the 16S rRNA gene sequence was used to assign these strains to the phyla, Firmicutes, Actinobacteria or Proteobacteria. Furthermore, most of the strains Firmicutes belonged to the order Lactobacillales. Interestingly, 12 of the isolated strains have not been previously reported from the Korean Peninsula. Also, based on their high 16S rRNA gene sequence similarities(>98.7%) and formation of strong monophyletic clades with the closest type species, each isolated strain of isolates was assigned to an independent, predefined bacterial species. Gram-stain reaction, colony and cell morphology, biochemical characteristics, isolation source, and NIBR IDs are described in the species description section.

주제어

표/그림 (6)

표 Table 1. Summary of the isolated strains from the gastrointestinal tract of the endangered species in Korea and their taxonomic affiliations.
그림 Fig. 1. Transmission electron micrographs of the isolated strains.
그림 Fig. 2. Phylogenetic tree based on 16S rRNA gene sequence comparisons, showing the relationship between the isolated strains in this study and the notable species from phylum Firmicutes (a) order Lactobacillales (In particular Enterococcus, Lactobacillus and Vagococcus), phylum Actinobacteria (c) and phylum Proteobacteria (d) and.
그림 Fig. 2. Continued.
그림 Fig. 2. Continued.
그림 Fig. 2. Continued.

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제안 방법

Biochemical analysis and gram-staining were performed using API kits (API 20NE, API ZYM and API ID 32GN; bioMerieux), GEN III microplates(Biolog) and a Gram-stain kit according to manufacturer’s instructions.

대상 데이터

Strain AE4-1 (=NIBRBAC000503069) was isolated from the feces of a green sea turtle (Chelonia mydas), Chungcheongnam Province, Korea.
Strain M1T305 ( =NIBRBAC000499685) was isolated from the feces of a Siberian musk deer (Moschus moschiferus), Hwacheon, Gangwon Province, Korea. The DNA G+C content of the type strain is 35.
Strain VM2501 (=NIBRBAC000499830) was isolated from the feces of an Andean condor (Vultur gryphus), Gwacheon, Gyeonggi Province, Korea. The DNA G+C content of the type strain is 67.
Strain VM3408 (=NIBRBAC000499831) was isolated from the feces of an Andean condor (Vultur gryphus), Gwacheon, Gyeonggi Province, Korea. The DNA G+C content of the type strain is 48.
Strain VT2412 (=NIBRBAC000499827) was isolated from the feces of an Andean condor (Vultur gryphus), Gwacheon, Gyeonggi Province, Korea. The DNA G+C content of the type strain is 65.
Twelve unreported bacterial strains were isolated from the gastrointestinal tract(GIT) and/or feces of various endangered species(green sea turtle, finless porpoise, Siberian musk deer, and Andean condor) found on the Korean Peninsula, which were collected by National Institute of Biological Resources and Seoul Grand Zoo. These isolates were assigned to the phyla Firmicutes, Bacteroidetes, or Proteobacteria, and of the Firmicutes, most belong to the order Lactobacillales(Table 1).

이론/모형

Homogenized GIT tissue and fecal samples were serially diluted with sterile phosphate buffered saline (PBS) and spread onto different culture media (Reasoner’s 2A Agar [R2A], Marine Agar 2216 [MA], Tryptic Soy Agar [TSA], and Brain heart infusion [BHI] agar], supplemented with 5% sheep blood, and incubated for at 25-30℃ for 1 week. Isolated, single bacterial colonies were prepared using the streak-plating method. The colony morphology and cell size were recorded after incubation on the appropriate growth media (R2A, TSA or MA) at 25-30℃ for 2 days.
The 16S rRNA gene sequences of the bacterial isolates were aligned with the corresponding sequences of bacterial reference strains using the BioEdit software with the multiple alignment algorithm [CLUSTAL W; (Thompson et al., 1994; Hall, 1999)]. Phylogenetic trees, using the 16S rRNA gene sequences of the isolates and the closely related bacterial species were constructed using the MEGA 7 software (Kumar et al.

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