Lipingzhangella rawalii sp. nov., a novel halophile isolated from Sambhar Salt Lake, Rajasthan, India.

A haloalkaliphilic actinobacterial strain LS1_29T, isolated from an inland hypersaline Sambhar Salt Lake, situated in Rajasthan, India, was subjected to taxonomic characterisation using the polyphasic approach. Cells of the strain were Gram stain positive and aerobic, having reticulate and aerial hyphae. The major fatty acids detected were iso C16:0 (38.23%), anteiso C17:0 (20.52%), iso C18:0 10 methyl (8.09%), iso C18:0 (7.74%) and iso C17:0 (7.48%). The major polar lipids identified were diphosphatidyl glycerol, phosphatidylglycerol, phosphatidylcholine and phospholipids. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the strain belonged to the genus Lipingzhangella with the highest similarity of 94.09% to Lipingzhangella halophila EGI 80537 T and formed a separate clade within the genus Lipingzhangella. The DNA G + C content of genomic DNA of strain LS1_29T was 67.99 mol%. The strain showed the highest orthologous average nucleotide identity (OrthoANI) value of 72.80% and digital DNA-DNA hybridization (dDDH) value of 20.3% with Lipingzhangella halophila EGI 80537 T. Based on the phenotypic, chemotaxonomic and phylogenetic characteristics, strain LS1_29T represents a novel species within the genus Lipingzhangella, for which the name Lipingzhangella rawalii sp. nov. is proposed. The type strain of the type species is LS1_29T (= KCTC 49199 T = JCM 32979 T = MCC 3420 T).

Genomic analysis of a novel species Halomonas shambharensis isolated from hypersaline lake in Northwest India.

Genome analysis of Halomonas shambharensis, a novel species, was performed to understand the osmoprotectant strategies used by the strain to overcome the salinity stress and to explore the prospective industrial uses. It will also help to better understand the ecological roles of Halomonas species in hypersaline habitats. Ultrastructure of the cell was determined by using transmission electron microscopy. Standard microbiological methods were used to find out growth parameters and heterotrophic mode of nutrition. For Genome analysis, complete bacterial genome sequencing was performed using the Oxford Nanopore MinION DNA Sequencer. Assembly, annotation and finishing of the obtained sequence were done by using a Prokaryotic Genome Annotation Pipeline (PGAP) (SPAdes v. 3.10.1). Predicted Coading sequences (CDSs) obtained through the PGAP were used for functional annotation using Clusters of Orthologous Groups and Kyoto Encyclopedia of Genes and Genomes (KEGG) platforms. The H. shambharensis was found to be a Gram-stain-negative, rod-shaped bacterium, motile with a peritrichous flagella. The H. shambharensis bacterium can grow in a wide range of temperature (from 25 to 65 °C), pH (pH 4 to pH 12.0) and salt concentration (5.0% NaCl to 30.0% NaCl). After annotation and assembly, the total genome size obtained was 1,533,947 bp, which revealed 146 subsystems, 3847 coding sequences, and 19RNAs with G+C content of 63.6%. Gene annotation identified the genes related to various metabolic pathways, including carbohydrate metabolism, fatty acid metabolism and stress tolerance. The genomic dataset of H. shambharensis will be useful for analysis of protein-coding gene families and how these coding genes are significant for the survival and metabolism among the different species of Halomonas. The complete genome sequence presented here will help to unravel the biotechnological potential of H. shambharensis for production of the high-value products such as betaine, or as a source of gene-mining for individual enzymes.

Betaine accumulation suppresses the de-novo synthesis of ectoine at a low osmotic concentration in Halomonas sp SBS 10, a bacterium with broad salinity tolerance.

The study aims to find out osmoadaptive mechanism used to overcome the salinity stress by Halomonas sp SBS 10 isolated from the saltern crystallizer ponds of the Sambhar Salt Lake and its taxonomic position using neighbor-joining algorithm. The strain SBS 10 was tested for accumulation of two major compatable solutes betaine and ectoine and was observed that osmoprotection in the strain SBS 10 is achieved by the accumulation of betaine or by the de-novo synthesis of betaine or ectoine. Amount of endogenous content of the betaine and ectoine per milligram of cell biomass was estimated to be 581 µg, 587 µg, 588 µg, 617 µg, and 761 µg for betaine and 1.52 µg, 2.74 µg, 3.14 µg, 3.50 µg, and 52.67 µg for ectoine, when exposed to 5, 10, 15, 20 and 25% of NaCl concentration. Results obtained from HPLC analysis showed that the betaine accumulation suppresses the de-novo synthesis of ectoine partially at low NaCl concentration in the growth medium. However, at a high NaCl concentration, the ectoine concentration increases abruptly as compared to the betaine. This indicates that the ectoine accumulation is transcriptionally up-regulated by the salinity stress. Phylogenetic analysis based on the neighbor-joining algorithm included the strain SBS 10 in the genus Halomonas of the family Halomonadaceae belonging to the class Gammaproteobacteria. Most closely related type strain was found to be Halomonas gudaonensis SL014B-69T (98.2% similarity). Ultrastructure characteristics showed the strain to be non-spore forming rod, 0.3-0.4 × 0.75-1.65 µm in size and motile with the help of peritrichous flagella.

Keratinophilic fungi from the vicinity of salt pan soils of Sambhar lake Rajasthan (India).

Forty soil samples were collected from seven sites in the vicinity of Sambhar lake Rajasthan, India and screened for the presence of keratinophilic fungi using hair baiting techniques for isolation. Seventeen isolates were recovered and identified. The cultures were identified by recognition of their macro- and micro- morphological features. Their identification was confirmed by BLAST using ITS1-5.8S-ITS2 rDNA region and sequences have been deposited in NCBI data base. A total of 34 species belonging to 29 genera were isolated. Among the dermatophytes and related keratinophilic fungi Chrysosporium indicum was predominant followed by Ctenomyces serratus, C. tropicum, Keratinophyton durus, Auxarthron conjugatum, Gymnascella dankaliensis, Gymnoascoideus petalosporus and Uncinocarpus reesii. Twenty-six species belonging to 22 genera represented other species. Our study indicates that keratinophilic fungi and species are found in the soils at the vicinity of the Sambhar Lake, and human activities can be the potential source of pathogenic fungi.