A Gram-positive, rod-shaped, endospore-forming Bacillus isolate, Bi.(FFUP1) (T), recovered in Portugal from a health product was subjected to a polyphasic study and compared with the type strains of Bacillus pumilus, Bacillus safensis, Bacillus altitudinis and Bacillus xiamenensis, the phenotypically and genotypically most closely related species. Acid production from cellobiose, D-glucose and D-mannose and absence of acid production from D-arabinose, erythritol, inositol, maltose, mannitol, raffinose, rhamnose, sorbitol, starch and L-tryptophan discriminated this new isolate from the type strains of the most closely related species. Additionally, a significant different protein and carbohydrate signature was evidenced by spectroscopic techniques, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and Fourier transform IR spectroscopy with attenuated total reflectance. Using a chemometric approach, the score plot generated by principal component analysis clearly delineated the isolate as a separate cluster. The quinone system for strain Bi.(FFUP1) (T) comprised predominantly menaquinone MK-7 and major polar lipids were diphosphatidylglycerol, an unidentified phospholipid and an unidentified glycolipid. Strain Bi.(FFUP1) (T) showed ? 99% 16S rRNA gene sequence similarity to B. safensis FO-036b(T), B. pumilus (7061(T) and SAFR-032), B. altitudinis 41KF2b(T) and B. xiamenensis HYC-10(T). Differences in strain Bi.FFUP1 (T) gyrB and rpoB sequences in comparison with the most closely related species and DNA-DNA hybridization experiments with Bi.FFUP1 (T) and B. pumilus ATCC 7061(T), B. safensis FO-036b(T), B. altitudinis 41KF2b(T) and B. xiamenensis HYC-10(T) gave relatedness values of 39.6% (reciprocal 38.0%), 49.9% (reciprocal 42.9%), 61.9% (reciprocal 52.2%) and 61.7% (reciprocal 49.2%), respectively, supported the delineation of strain Bi.(FFUP1) (T) as a representative of a novel species of the genus Bacillus, for which the name Bacillus invictae sp. nov. is proposed, with strain Bi.(FFUP1) (T) (=DSM 26896(T) =CCUG 64113(T)) as the type strain.

The aim of this study was to reclarify the taxonomic status of strain Bacillus invictae Bi.FFUP1T by performing comparative analyses with the other four type strains within the Bacillus pumilus group. The digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) values between strains B. invictae Bi.FFUP1T (= DSMZ 26896T = MCCC 1A07089T), B. altitudinis 41KF2bT (= DSMZ 21631T = MCCC 1A06452T), B. safensis FO-36bT (= DSMZ 19292T = MCCC 1A6451T), B. pumilus ATCC 7061T (= DSMZ 27T = MCCC 1A06453T) and B. xiamenensis HYC-10T (= MCCC 1A00008T) were, respectively, 82.90 % and 98.10 %, which are greater than the thresholds for bacterial species delineation, suggesting that they should belong to the same species, while the dDDH and ANI values between strain B. invictae DSMZ 26896T and the other three type strains within the B. pumilus group were below the respective thresholds of 70 % and 95 %. Meanwhile, B. invictae DSMZ 26896T and B. altitudinis 41KF2bT shared 98.7 % gyrB gene sequence similarity based on resequencing, whereas strain B. invictae DSMZ 26896T shared low similarities (< 95 %) with the other three type strains. In addition, in comparison with those from the other three type strains, phenotypic data of B. invictae DSMZ 26896T and B. altitudinis 41KF2bT, including API 20NE, API ZYM, Biolog GN2 and API 50CHB tests, showed slight differences. The data from these combined genotypic and phenotypic analyses suggest that Bacillus invictae Branquinho et al. 2014 should be regarded as a later heterotypic synonym of Bacillus altitudinisShivaji et al. 2006.

A novel bacterial strain NIO-1130(T) was isolated from sediment sample taken from Chorao Island, Goa Province, India, and subjected to a taxonomic investigation. The strain was Gram-positive, aerobic, and motile. Phylogenetic analysis based on 16S rRNA gene sequences placed the isolate within the genus Bacillus and strain NIO-1130(T) showed highest sequence similarity with Bacillus halosaccharovorans DSM 25387(T) (98.4%) and Bacillus niabensis CIP 109816(T) (98.1%), whereas other Bacillus species showed <97.0% similarity. Tree based on gyrB gene sequence revealed that strain bacillus group. The major menaquinone was MK-7 and the predominant cellular fatty acids were iso-C15:0, anteiso-C15:0, iso-C17:0, and anteiso-C17:0. The strain showed a DNA G+C content of 39.9 mol%. DNA-DNA hybridization studies revealed that strain NIO-1130(T) exhibits 70% similarity with Bacillus halosaccharovorans DSM 25387(T) and Bacillus niabensis CIP 109816(T). On the basis of physiological, biochemical, chemotaxonomic and phylogenetic analyses, we consider the isolate to represent a novel species of the genus Bacillus, for which the name Bacillus cellulasensis sp. nov., is proposed. The type strain is NIO-1130(T) (=NCIM 5461(T)=CCTCC AB 2011126(T)).

In the study by Mawlankar et al. in Arch Microbiol 198:83-89 (2016), the phylogenetic position of strain "Bacillus cellulasensis" NIO-1130(T) based on 16S rRNA and gyrB genes was inconsistent. Therefore, the aim of this study is to re-determine its taxonomic status using diverse genotypic approaches including single gene analysis, multilocus sequence analysis, and genomic analyses. The reconstructed phylogenetic trees based on 16S rRNA gene and six concatenated genes showed that "B. cellulasensis" NIO-1130(T) (=NCIM 5461(T) = CCTCC AB 2011126(T)) revealed the closest genetic relationship with type strain Bacillus altitudinis 41KF2b(T), with 98.6-100 % similarities of 16S rRNA gene, gyrB, pycA, pyrE, mutL, aroE, trpB, and six concatenated housekeeping genes. The high similarities for gene(s) sequences between "B. cellulasensis" NIO-1130(T) and B. altitudinis 41KF2b(T) indicated that they should be conspecific. The DNA G+C content for strain NIO-1130(T) was determined to be 41.3 mol% and identical to that of B. altitudinis 41KF2b(T). Moreover, 88.4 % of digital DNA-DNA hybridization and 98.7 % of average nucleotide identity values between two strains were much higher than the standard criteria for delineation of bacterial species, suggesting that they belonged to the same species. Therefore, the data from the combined genotypic analyses suggest that "Bacillus cellulasensis" should be classified as a member of Bacillus altitudinis.