Please cite this article in press as: Y. Shivani, et al., Spirochaeta lutea sp. nov., isolated from marine habitats and emended description of the genus Spirochaeta, Syst. Appl. Microbiol. (2014), http://dx.doi.org/10.1016/j.syapm.2014.11.002 ARTICLE IN PRESS G Model SYAPM-25663; No. of Pages 5 Systematic and Applied Microbiology xxx (2014) xxx–xxx Contents lists available at ScienceDirect Systematic and Applied Microbiology j ourna l h omepage: www.elsevier.de/syapm Spirochaeta lutea sp. nov., isolated from marine habitats and emended description of the genus Spirochaeta  Y. Shivani a , Y. Subhash b , L. Tushar b , Ch. Sasikala a , Ch.V. Ramana b,∗ a Bacterial Discovery Laboratory, Centre for Environment, Institute of Science and Technology, J. N. T. University, Kukatpally, Hyderabad 500 085, India b Department of Plant Sciences, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad 500 046, India a r t i c l e i n f o Article history: Received 15 July 2014 Received in revised form 5 November 2014 Accepted 17 November 2014 Keywords: Spirochaeta lutea Spirochaetes Whole genome sequencing Chemotaxonomic analysis a b s t r a c t Metagenome analysis of forty samples indicated a rich diversity of free living spirochaetes among Indian habitats. Only 2–3% of the total bacterial diversity was represented by spirochaetes with five distinct phylogenetic OTUs. A strain (JC230 T ) was brought to cultivation and its 16S rRNA gene sequence showed highest sequence similarity with that of Spirochaeta asiatica Z-7591 T (90.78%). Genome sequence analysis of strain JC230 T indicates the presence of 117 genes related to motility and chemotaxis. Strain JC230 T is an obligate anaerobe. It is helical shaped, Gram-stain-negative and catalase and oxidase negative. Strain JC230 T produces carotenoids and have all the genes related to carotenogenesis of spirilloxanthin series. Isoprenoid biosynthesis occurs through HMG-CoA (3-hydroxy-3-methylglutaryl-coenzyme A) pathway. Twelve bacterial haemoglobin and one flavohaemoglobin related genes are present in the genome of strain JC230 T . The genome of strain JC230 T showed the presence of heavy metal resistance related genes of cobalt, zinc, cadmium and arsenic. Six genes related to colicin V and bacteriocin production cluster are present. Major (>2%) fatty acids were C 12:0 , C 14:0 , iso-C 14:0 , iso-C 15:1 H/C 13:0 3OH, anteiso-C 15:0 , C 16:0 , and iso-C 16:0 . Strain JC230 T contains diphosphatylglycerol, phosphatydilglycerol, phosphatydilethanolamine, unidentified phospholipid (PL1) and unidentified lipids (L1-7). G + C mol% of strain JC230 T was 54.1%. Dis- tinct morphological, physiological and genotypic differences from the previously described taxa support the classification of strain JC230 T as a representative of a new species in the genus Spirochaeta, for which the name Spirochaeta lutea sp. nov. is proposed. The type strain is JC230 T (=KCTC 15387 T = DSM 29074 T ). © 2014 Elsevier GmbH. All rights reserved. The genus Spirochaeta is represented by a large number of free living non-pathogenic, helical shaped bacteria commonly found in the anoxic mud, marshes, marine sediments, soda lakes and microbial mats [1,2,4,8,9,13,17,25]. The genus Spirochaeta cur- rently has 17 validly published species names (including the recently described Spirochaeta psychrophila [16]) of which, genome sequences for 9 type strains are available. Only one species name added to the genus Spirochaeta was based on a strain isolated from India [26]. During a survey of free living spirochaetes, we have ana- lyzed a large number of samples from a few coastal habitats of India and isolated an obligately anaerobic spirochaete, designated it as strain JC230 T which was characterized using polyphasic taxonomy and through genome sequence.  The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequence and the draft genome sequence of strain JC230 T are HG965770 and JNUP00000000 (version JNUP01000000), respectively. ∗ Corresponding author. Tel.: +91 8179724006. E-mail addresses: sasi449@yahoo.ie (Ch. Sasikala), cvr449@yahoo.com, cvr449@gmail.com (Ch.V. Ramana). Soil samples collected from Gujarat (GPS positioning of the sample collection site; 23 ◦ 8 ′ 82 ′′ N and 69 ◦ 74 ′ E), Tamil Nadu (GPS positioning of the sample collection site is 9 ◦ 06 ′ 24 ′′ N, 79 ◦ 31 ′ 28 ′′ E; 8 ◦ 04 ′ 42 ′′ N, 77 ◦ 32 ′ 30 ′′ E) during December, 2012 were used for metagenome analysis. A total of 40 composite samples were made from the samples collected and soil DNA was extracted from all the samples using HiPurA TM soil DNA isolation kit (Himedia Lab- oratories Pvt. Ltd, Mumbai, India) according to the manufacturer’s protocol, with 0.25 g of soil (dry wt.). An average of 7 ng of DNA was recovered from 1 g of soil sample. The extracted DNA sam- ples were stored at -20 ◦ C. DNA samples were sent for bacterial tag-encoded FLX 454 pyrosequencing to Research and Testing Lab- oratory LLC (Lubbock, TX, USA). The samples underwent partial amplification of the V1–V3 region of the 16S rRNA gene as described previously [6]. The sequence data obtained after pyrosequencing were processed as described previously [7,19,20]. All taxonomic classification was performed using Mothur’s version of the RDP Bayesian classifier, using a RDP training dataset number 9 (available at http://www.mothur.org/wiki/RDP reference files). All sequence data are available under the following NCBI SRA accession nos. SRP034851, SRP040491 and SRP034851. http://dx.doi.org/10.1016/j.syapm.2014.11.002 0723-2020/© 2014 Elsevier GmbH. All rights reserved.