Comparative Genome Analysis of Two Bacillus pumilus Strains Producing High Level of Extracellular Hydrolases.

Whole-genome sequencing of a soil isolate Bacillus pumilus, strain 7P, and its streptomycin-resistant derivative, B. pumilus 3-19, showed genome sizes of 3,609,117 bp and 3,609,444 bp, respectively. Annotation of the genome showed 3794 CDS (3204 with predicted function) and 3746 CDS (3173 with predicted function) in the genome of strains 7P and 3-19, respectively. In the genomes of both strains, the prophage regions Bp1 and Bp2 were identified. These include 52 ORF of prophage proteins in the Bp1 region and 38 prophages ORF in the Bp2 region. Interestingly, more than 50% of Bp1 prophage proteins are similar to the proteins of the phi105 in B. subtilis. The DNA region of Bp2 has 15% similarity to the DNA of the Brevibacillus Jimmer phage. Degradome analysis of the genome of both strains revealed 148 proteases of various classes. These include 60 serine proteases, 48 metalloproteases, 26 cysteine proteases, 4 aspartate proteases, 2 asparagine proteases, 3 threonine proteases, and 2 unclassified proteases. Likewise, three inhibitors of proteolytic enzymes were found. Comparative analysis of variants in the genomes of strains 7P and 3-19 showed the presence of 81 nucleotide variants in the genome 3-19. Among them, the missense mutations in the rpsL, comA, spo0F genes and in the upstream region of the srlR gene were revealed. These nucleotide polymorphisms may have affected the streptomycin resistance and overproduction of extracellular hydrolases of the 3-19 strain. Finally, a plasmid DNA was found in strain 7P, which is lost in its derivative, strain 3-19. This plasmid contains five coding DNA sequencing (CDS), two regulatory proteins and three hypothetical proteins.

Supplementation of Bacillus subtilis GM5 enhances broiler body weight gain and modulates cecal microbiota.

We investigated the effect of the strain Bacillus subtilis GM5 on growth, feed conversion, and the composition of cecum microbiota in broiler chickens. Half of which received a control diet, while the other half was fed a diet supplemented with GM5 spores. Cecal contents on days 1, 10, and 42 were subjected to metataxonomic analysis. Principal Component Analysis showed that the control and probiotic groups formed three separate clusters, indicating changes, which occurred gradually in microbial communities. On day 1, Firmicutes (53.87-57.61%) and Proteobacteria (43.77-38.93%) were prevalent in both groups, whereas samples of days 10 and 42 were predominantly occupied by Firmicutes (54.55-81.79%) and Bacteroidetes (26.94-30.45%). In the group of chickens treated with probiotic, the average daily gain in body weight was higher, while feed conversion decreased by 1.44%. A surge in the presence of beneficial bacteria of the Ruminococcaceae family was observed. The introduction of the probiotic led to an elevated Firmicutes/Bacteroidetes ratio, which positively correlated with chickens' bodyweight (Spearman ρ = 1.0, P < 0.05). Supplementing broiler feed with B. subtilis GM5 spores leads to improved feed intake and digestibility, which is paramount in reducing the cost of the final product. Thus, the probiotic strain GM5 modulates the cecal microbiota of broiler chickens and increases microbial diversity, which is well exhibited on the 42nd day. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02634-2.

Structure and variation of root-associated microbiomes of potato grown in alfisol.

Root-associated fungi and bacteria play a pivotal role in the plant-soil ecosystem by influencing both plant growth and immunity. The aim of this study was to unravel the biodiversity of the bacterial and fungal rhizosphere (RS) and rhizoplane (RP) microbiota of Zhukovskij rannij potato (Solanum tuberosum L.) cultivar growing in the Alfisol of Tatarstan, Russia. To assess the structure and diversity of microbial communities, we employed the 16S rRNA and internal transcribed spacer gene library technique. Overall, sequence analysis showed the presence of 3982 bacterial and 188 fungal operational taxonomic units (OTUs) in the RP, and 6018 bacterial and 320 fungal OTUs for in the RS. Comparison between microbial community structures in the RS and RP showed significant differences between these compartments. Biodiversity was higher in the RS than in the RP. Although members of Proteobacteria (RS-59.1 ± 4.9%; RP-54.5 ± 9.2%), Bacteroidetes (RS-23.19 ± 10.2%; RP-34.52 ± 10.4%) and Actinobacteria (RS-11.55 ± 4.9%; RP-7.7 ± 5.1%) were the three most dominant phyla, accounting for 94-98% of all bacterial taxa in both compartments, notable variations were observed in the primary dominance of classes and genera in RS and RP samples. In addition, our results demonstrated that the potato rhizoplane was significantly enriched with the genera Flavobacterium, Pseudomonas, Acinetobacter and other potentially beneficial bacteria. The fungal community was predominantly inhabited by members of the Ascomycota phylum (RS-81.4 ± 8.1%; RP-81.7 ± 5.7%), among which the genera Fusarium (RS-10.34 ± 3.41%; RP-9.96 ± 4.79%), Monographella (RS-7.66 ± 4.43%; RP-9.91 ± 5.87%), Verticillium (RS-4.6 ± 1.43%; RP-8.27 ± 3.63%) and Chaetomium (RS-4.95 ± 2.07%; RP-8.33 ± 4.93%) were particularly abundant. Interestingly, potato rhizoplane was significantly enriched with potentially useful fungal genera, such as Mortierella and Metacordiceps. A comparative analysis revealed that the abundance of Fusarium (a cosmopolitan plant pathogen) varied significantly depending on rotation variants, indicating a possible control of phytopathogenic fungi via management-induced shifts through crop rotational methods. Analysis of the core microbiome of bacterial and fungal community structure showed that the formation of bacterial microbiota in the rhizosphere and rhizoplane is dependent on the host plant.

Comparative Genome Analysis of Uropathogenic Morganella morganii Strains.

Morganella morganii is an opportunistic bacterial pathogen shown to cause a wide range of clinical and community-acquired infections. This study was aimed at sequencing and comparing the genomes of three M. morganii strains isolated from the urine samples of patients with community-acquired urinary tract infections. Draft genome sequencing was conducted using the Illumina HiSeq platform. The genomes of MM 1, MM 4, and MM 190 strains have a size of 3.82-3.97 Mb and a GC content of 50.9-51%. Protein-coding sequences (CDS) represent 96.1% of the genomes, RNAs are encoded by 2.7% of genes and pseudogenes account for 1.2% of the genomes. The pan-genome containes 4,038 CDS, of which 3,279 represent core genes. Six to ten prophages and 21-33 genomic islands were identified in the genomes of MM 1, MM 4, and MM 190. More than 30 genes encode capsular biosynthesis proteins, an average of 60 genes encode motility and chemotaxis proteins, and about 70 genes are associated with fimbrial biogenesis and adhesion. We determined that all strains contained urease gene cluster ureABCEFGD and had a urease activity. Both MM 4 and MM 190 strains are capable of hemolysis and their activity correlates well with a cytotoxicity level on T-24 bladder carcinoma cells. These activities were associated with expression of RTX toxin gene hlyA, which was introduced into the genomes by a phage similar to Salmonella phage 118970_sal4.

Draft genome sequence and analysis of Klebsiella oxytoca strain NK-1 isolated from ureteral stent.

Klebsiella oxytoca is a facultative aerobic, gram-negative, rod-shaped bacterium capable of causing nosocomial infections, in particular catheter-associated urinary tract infections (CAUTIs). Data on the possible roles of uncommon pathogens such as K. oxytoca in the pathogenesis of biofilm-associated infections such as CAUTIs have been already reported. Herein, we describe the draft genome sequence of K. oxytoca strain NK-1 isolated from the surface of ureteral stent retrieved from a Russian female. The genome comprises 6,232,464 bp, with a G + C content of 55.60% and an L 50 of 7. A total of 6246 putative protein-encoding genes were predicted, including considerable number of genes responsible for adhesion, invasion, drug resistance, iron acquisition and other genes relevant for virulence. The NK-1 strain was ascribed a sequence type (ST) as ST 216 (4, 6, 19, 10, 46, 24, 31). Data comparison of the recA gene sequences confirmed that the strain belongs to the species K. oxytoca. Minimal inhibitory concentration of different antibiotics have been determined. This whole genome shotgun project has been deposited at DDBJ/ENA/GenBank under the accession number QPKC00000000.1.

Draft genome sequence data of Lysinibacillus fusiformis strain GM, isolated from potato phyllosphere as a potential probiotic.

Here we present the morphological and physiological properties of isolated Lysinibacillus fusiformis strain GM, its draft genome sequence as well as annotation and analysis of its genome. Initial analysis of MALDI-TOF mass spectrometry, 16S rRNA gene analysis and in silico DNA-DNA hybridization revealed that the strain belongs to the species Lysinibacillus fusiformis. The 4,678,122 bp draft genome consist of 17 scaffolds encoding 4588 proteins and 137 RNAs. Annotation of the genome sequence revealed cellulase and protease encoding genes, genes of adhesion proteins and putative genes responsible for the biosynthesis of antimicrobial metabolites. The Whole Genome Shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number NTMQ00000000.1 (https://www.ncbi.nlm.nih.gov/nuccore/NZ_NTMQ00000000.1).