1887

Abstract

A novel Gram-stain-positive, strictly aerobic, short rod-shaped bacterium, designated 2C, was isolated from freshly packaged microfiltered milk. This strain was able to grow within the NaCl concentration range of 0–5 % (w/v), temperature range of 8–37 °C (optimally at 30 °C) and at pH 6.0–10.0. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain 2C was closely related to species of the genus , with the highest sequence similarity (99.2 %) to DSM 20427 as well as DSM 18909 (=YM18-098). The phylogenetic tree based on 16S rRNA genes showed that strain 2C clustered with DSM 18909. However, the phylogenetic tree based on concatenated 16S rRNA and four housekeeping genes showed that strain 2C clustered with DSM 20427. Furthermore, the phylogenomic tree showed that strain 2C clustered with DSM 20427 and DSM 18909. The major respiratory quinones were MK-10, MK-11 and MK-12. The predominant cellular fatty acids were anteiso-C, iso-C and anteiso-C. The polar lipid composition of strain 2C consisted of diphosphatidylglycerol, phosphatidylglycerol, three unidentified glycolipids and two unidentified lipids. The cell-wall peptidoglycan type was a variant of B1 {Gly} [-Lys] -Glu--Lys, with the amino acids lysine, glycine, alanine and glutamic acid. The whole-cell sugars consisted of galactose, glucose, ribose and minor amounts of rhamnose. In addition, strain 2C showed a glycolyl-type cell wall. The genomic DNA G+C content was 69.8mol%, while the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values with the closely related species were below the recognized thresholds of 95–96 % ANI and 70 % DDH for species definition. Based on the phenotypic and genotypic data, strain 2C (=LMG 32277=CECT 30329) is considered to represent a new species, for which the name sp. nov. is proposed.

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2021-11-22
2024-04-19
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References

  1. Orla-Jensen S. The Lactic Acid Bacteria Høst: Copenhagen; 1919 pp 1–196
    [Google Scholar]
  2. Suzuki K, Hamada M. Microbacterium. In Bergey’s Manual of Systematics of Archaea and Bacteria Wiley; 2015 pp 1–52 [View Article]
    [Google Scholar]
  3. Salam N, Jiao J-Y, Zhang X-T, Li W-J. Update on the classification of higher ranks in the phylum Actinobacteria. Int J Syst Evol Microbiol 2020; 70:1331–1355 [View Article]
    [Google Scholar]
  4. Zlamala C, Schumann P, Kämpfer P, Valens M, Rosselló-Mora R et al. Microbacterium aerolatum sp. nov., isolated from the air in the’Virgilkapelle’in Vienna. Int J Syst Evol Microbiol 2002; 52:1229–1234 [View Article]
    [Google Scholar]
  5. Vaz-Moreira I, Lopes AR, Falsen E, Schumann P, Nunes OC et al. Microbacterium luticocti sp. nov., isolated from sewage sludge compost. Int J Syst Evol Microbiol 2008; 58:1700–1704 [View Article]
    [Google Scholar]
  6. Vaz-Moreira I, Lopes AR, Faria C, Sproeer C, Schumann P et al. Microbacterium invictum sp. nov., isolated from homemade compost. Int J Syst Evol Microbiol 2009; 59:2036–2041 [View Article]
    [Google Scholar]
  7. Chen X, Li G-D, Li Q-Y, Xu F-J, Jiang C-L et al. Microbacterium faecale sp. nov., isolated from the faeces of Columba livia. Int J Syst Evol Microbiol 2016; 66:4445–4450 [View Article]
    [Google Scholar]
  8. Chen X, Li Q-Y, Li G-D, Xu F-J, Jiang Y et al. Microbacterium gilvum sp. nov., isolated from civet faeces. Antonie van Leeuwenhoek 2016; 109:1177–1183 [View Article]
    [Google Scholar]
  9. Bellassi P, Cappa F, Fontana A, Morelli L. Phenotypic and genotypic investigation of two representative strains of Microbacterium species isolated from micro-filtered milk: growth capacity and spoilage-potential assessment. Front Microbiol 2020; 11:2555 [View Article]
    [Google Scholar]
  10. Madhaiyan M, Poonguzhali S, Lee J-S, Lee K-C, Saravanan VS et al. Microbacterium azadirachtae sp. nov., a plant-growth-promoting Actinobacterium isolated from the rhizoplane of neem seedlings. Int J Syst Evol Microbiol 2010; 60:1687–1692 [View Article]
    [Google Scholar]
  11. Gao M, Wang M, Zhang Y-C, Zou X-L, Xie L-Q et al. Microbacterium neimengense sp. nov., isolated from the rhizosphere of maize. Int J Syst Evol Microbiol 2013; 63:236–240 [View Article]
    [Google Scholar]
  12. Yan Z-F, Lin P, Won K-H, Yang J-E, Li C-T et al. Microbacterium hibisci sp. nov., isolated from rhizosphere of mugunghwa (Hibiscus syriacus L.). Int J Syst Evol Microbiol 2017; 67:3564–3569 [View Article]
    [Google Scholar]
  13. Kim KK, Lee KC, Oh H-M, Lee J-S. Microbacterium aquimaris sp. nov., isolated from seawater. Int J Syst Evol Microbiol 2008; 58:1616–1620 [View Article]
    [Google Scholar]
  14. Mawlankar RR, Mual P, Sonalkar VV, Thorat MN, Verma A et al. Microbacterium enclense sp. nov., isolated from sediment sample. Int J Syst Evol Microbiol 2015; 65:2064–2070 [View Article]
    [Google Scholar]
  15. Kook M, Son H-M, Yi T-H. Microbacterium kyungheense sp. nov. and Microbacterium jejuense sp. nov., isolated from salty soil. Int J Syst Evol Microbiol 2014; 64:2267–2273 [View Article]
    [Google Scholar]
  16. Lee I, Chalita M, Ha SM, Na SI, Yoon SH et al. ContEst16S: An algorithm that identifies contaminated prokaryotic genomes using 16S RNA gene sequences. Int J Syst Evol Microbiol 2017; 67:2053–2057 [View Article]
    [Google Scholar]
  17. Chun J, Oren A, Ventosa A, Christensen H, Arahal DR et al. Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 2018; 68:461–466 [View Article]
    [Google Scholar]
  18. Yoon S-H, Ha S-M, Kwon S, Lim J, Kim Y et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol 2017; 67:1613 [View Article]
    [Google Scholar]
  19. Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Mol Biol Evol 2018; 35:1547–1549 [View Article]
    [Google Scholar]
  20. Fidalgo C, Riesco R, Henriques I, Trujillo ME, Alves A. Microbacterium diaminobutyricum sp. nov., isolated from Halimione portulacoides, which contains diaminobutyric acid in its cell wall, and emended description of the genus Microbacterium. Int J Syst Evol Microbiol 2016; 66:4492–4500 [View Article]
    [Google Scholar]
  21. Seemann T. Prokka: Rapid prokaryotic genome annotation. Bioinformatics 2014; 30:2068–2069 [View Article]
    [Google Scholar]
  22. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994; 22:4673–4680 [View Article]
    [Google Scholar]
  23. Kim OS, Cho YJ, Lee K, Yoon SH, Kim M et al. Introducing EzTaxon-e: A prokaryotic 16s rRNA gene sequence database with phylotypes that represent uncultured species. Int J Syst Evol Microbiol 2012; 62:716–721 [View Article]
    [Google Scholar]
  24. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M et al. SPAdes: A new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012; 19:455–477 [View Article]
    [Google Scholar]
  25. Gurevich A, Saveliev V, Vyahhi N, Tesler G. QUAST: quality assessment tool for genome assemblies. Bioinformatics 2013; 29:1072–1075 [View Article]
    [Google Scholar]
  26. Parks DH, Imelfort M, Skennerton CT, Hugenholtz P, Tyson GW. CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes. Genome Res 2015; 25:1043–1055 [View Article]
    [Google Scholar]
  27. Richter M, Rosselló-Móra R. Shifting the genomic gold standard for the prokaryotic species definition. Proc Natl Acad Sci USA 2009; 106:19126–19131 [View Article]
    [Google Scholar]
  28. Auch AF, von Jan M, Klenk H-P, Göker M. Digital DNA-DNA hybridization for microbial species delineation by means of genome-to-genome sequence comparison. Stand Genomic Sci 2010; 2:117–134 [View Article]
    [Google Scholar]
  29. Meier-Kolthoff JP, Göker M, Spröer C, Klenk H-P. When should a DDH experiment be mandatory in microbial taxonomy?. Arch Microbiol 2013; 195:413–418 [View Article]
    [Google Scholar]
  30. Meier-Kolthoff JP, Auch AF, Klenk H-P, Göker M. Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 2013; 14:60 [View Article]
    [Google Scholar]
  31. Page AJ, Cummins CA, Hunt M, Wong VK, Reuter S et al. Roary: Rapid large-scale prokaryote pan genome analysis. Bioinformatics 2015; 31:3691–3693 [View Article]
    [Google Scholar]
  32. Belahbib H, Summers ZM, Fardeau ML, Joseph M, Tamburini C et al. Towards a congruent reclassification and nomenclature of the thermophilic species of the genus Pseudothermotoga within the order Thermotogales. Syst Appl Microbiol 2018; 41:555–563 [View Article]
    [Google Scholar]
  33. Darriba D, Posada D, Kozlov AM, Stamatakis A, Morel B et al. ModelTest-NG: a new and scalable tool for the selection of DNA and protein evolutionary models. Mol Biol Evol 2020; 37:291–294 [View Article]
    [Google Scholar]
  34. Bassi D, Cappa F, Cocconcelli PS. A combination of a SEM technique and X-ray microanalysis for studying the spore germination process of Clostridium tyrobutyricum. Res Microbiol 2009; 160:322–329 [View Article]
    [Google Scholar]
  35. Claus D. A standardized Gram staining procedure. World J Microbiol Biotechnol 1992; 8:451–452 [View Article]
    [Google Scholar]
  36. Tittsler RP, Sandholzer LA. The use of semi-solid agar for the detection of bacterial motility. J Bacteriol 1936; 31:575–580 [View Article]
    [Google Scholar]
  37. Anagnostopoulos GD, Seaman A, Woodbine M. An attempt to increase the heat resistance of Microbacterium lacticum. J Appl Bacteriol 1966; 29:207–212 [View Article]
    [Google Scholar]
  38. Bulut S, Waites WM, Mitchell JR. Effects of combined shear and thermal forces on destruction of Microbacterium lacticum. Appl Environ Microbiol 1999; 65:4464–4469 [View Article]
    [Google Scholar]
  39. Robinson K. Some observations on the taxonomy of the genus Microbacterium. I. Cultural and physiological reactions and heat resistance. J Appl Bacteriol 1966; 29:607–615 [View Article]
    [Google Scholar]
  40. Sasser M. Identification of bacteria by gas chromatography of cellular fatty acids. Tech Note 2001; 101:1–6 [View Article]
    [Google Scholar]
  41. Minnikin DE, O’Donnell AG, Goodfellow M, Alderson G, Athalye M et al. An integrated procedure for the extraction of bacterial isoprenoid quinones and polar lipids. J Microbiol Methods 1984; 2:233–241 [View Article]
    [Google Scholar]
  42. Hamada M, Yamamura H, Komukai C, Tamura T, Suzuki KI et al. Luteimicrobium album sp. nov., a novel Actinobacterium isolated from a lichen collected in Japan, and emended description of the genus Luteimicrobium. J Antibiot 2012; 65:427–431 [View Article]
    [Google Scholar]
  43. Uchida K, Kudo T, Suzuki KI, Nakase T. A new rapid method of glycolate test by diethyl ether extraction, which is applicable to a small amount of bacterial cells of less than one milligram. J Gen Appl Microbiol 1999; 45:49–56 [View Article]
    [Google Scholar]
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