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Introduction. The study of the ability of Bacillus anthracis strains with different phenotypic properties to spore germination, reproduction and sporulation on a medium based on an aqueous soil extract can help assess the significance of these processes in the formation and maintenance of soil anthrax foci. Aim. The analysis of individual characteristics of the development of a vegetative culture of anthrax pathogen strains with different phenotypes in a soil medium model. Materials and methods. On a group of anthrax microbe strains with different plasmid composition and virulence, the possibility of spore germination, reproduction of bacilli and, at least in some of them, productive spore formation on the soil medium was studied. Results. Three variants of culture development of B. anthracis strains were identified: 1 spores remain intact, not germinating; 2 after germination of spores, bacilli are formed, which multiply with different intensity, passing into involutional forms without spore formation; 3 the passage of a complete physiological cycle "sporebacillusspore". The presence of 2% blood in the soil environment contributed to the germination of spores and reproduction of the culture, but inhibited the process of sporulation during the observation period of 3 days. No correlation was found between a certain phenotype of the studied strains of B. anthracis and the ability to germinate and vegetate on soil media. Conclusion. The data obtained that only 17% of CFU gives rise to the formation of colonies on the soil medium suggest the heterogeneity of the properties of the population of the studied strains. Isolation of such cultures and their further detailed study will make it possible to identify the most significant complexes of biological properties for the realization of a complete physiological cycle under soil-simulating conditions.

Clostridioides difficile (C. difficile) is a Gram-positive, spore-forming, toxin-producing, obligate anaerobic bacterium. C. difficile infection (CDI) is the leading cause of healthcare-associated infective diarrhoea. The infection is mediated by the spore, a metabolically inactive form of C. difficile. The spore coat acts as a physical barrier to defend against chemical insults from hosts and natural environments. The composition of spore coat has already been revealed; therefore, the interactive networks of spore coat proteins and the dynamic process of coat assembly are the keys to design strategies to control and cure CDI. This review gives a brief discussion of the signal processing and transcriptional regulation of C. difficile sporulation initiation. Following the discussion, the spore formation is also introduced. Finally, this review mainly focuses on the spore coat assembly, a poorly understood process in C. difficile, and important proteins that have been studied.

Introduction. The study of the protein composition of the causative agent of anthrax — Bacillus anthracis, allows you to identify both general species and individual characteristics of strains that differ in phenotypic properties, manifested mainly in the vegetative form and which are important for virulence, immunogenicity and the ability to adapt to different vegetation conditions.Purpose of the work. In the group of anthrax microbe strains having different plasmid composition and virulence, different methods of extraction of the total proteome from vegetative bacillus cells have been tested.Results. In the course of the work, it was shown that the rate of spore formation varies significantly between individual strains of the anthrax microbe and can have a significant impact on the efficiency of extraction and the composition of the protein complex. Preliminary treatment with lysozyme, which affects the cell membrane, promotes a more complete lysis of cells, and ultramicrocentrifuge filtration provides complete specific sterility of the obtained samples.Conclusion. A culture preparation scheme was developed for B. anthracis, which allows one to obtain a culture in the vegetative phase of the life cycle and to efficiently extract proteins in combination with reliable disinfection of samples.

Streptomyces griseus S4-7, a well-characterized keystone taxon among strawberry microbial communities, shows exceptional disease-preventing ability. The whole-genome sequence, functional genes, and bioactive secondary metabolites of the strain have been described in previous studies. However, proteomics studies of not only the S4-7 strain, but also the Streptomyces genus as a whole, remain limited to date. Therefore, in the present study, we created a proteomics reference map for S. griseus S4-7. Additionally, analysis of differentially expressed proteins was performed against a wblE2 mutant, which was deficient in spore chain development and did not express an antifungal activity-regulatory transcription factor. We believe that our data provide a foundation for further in-depth studies of functional keystone taxa of the phytobiome and elucidation of the mechanisms underlying plant-microbe interactions, especially those involving the Streptomyces genus.

Ulva prolifera, the main causative species of green tide, has drawn much attention. Spore reproduction was one of the main reproduction strategies that could be induced by changing environmental factors, but the mechanism of spore formation remains obscure. Here, we cultured U. prolifera (segments) under the optimal sporulation condition, and four states in total from vegetative to reproductive were defined. Additionally, the chlorophyll fluorescence and transcriptome analysis were determined at these four states. The results showed that: (1) Compared with state I, the photosynthetic capacity (Fv/Fm, Fv′/Fm′, rETRmax) and chlorophyll content (Chl a, Chl b, carotenoids) were enhanced in state II, whereas it decreased in state III and IV (the spore formation period); (2) a total of 41,058 unigenes were expressed during the spore formation process; (3) compared with state I, the genes related with photosynthesis, terpenoid backbone biosynthesis, and carotenoid biosynthesis were significantly upregulated in states II, III, and IV whereas glycolysis was downregulated in state I; (4) some genes of the transcription factors families, such as the C3H family, may be one of the key factors that regulate genes in the spore formation; (5) 574 of the differentially expressed genes (DEGs) associated with flagella biosynthesis were annotated according to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes Orthology, including 20 unigenes encoding intraflagellar transport proteins (IFTs) which had not been reported in previous transcriptome analysis in U. prolifera. This study provides a new perspective of spore formation at the gene transcriptional level, although the detailed transcription regulatory spore formation network remains to be unveiled.

A decrease in therapeutic effect of some live lacto- and bifdobacteria-based drugs for veterinary use has been observed for the last 20 years that urges scientists to search for new microorganisms possessing probiotic properties. Many studies in this feld are focused on Bacillus subtilis that is widespread in the environment and non-pathogenic for animals and humans. Results of tests of Bacillus subtilis for its biological properties and antagonistic activity aimed at optimization of methodical approaches for detection of strain with the highest antagonistic effect on some opportunistic microorganisms and their further use as probiotics are described. Cultural morphological and biochemical characteristics of the tested strains conformed to the species characteristics of Bacillus subtilis. Tested strains were nonpathogenic for white mice. Tests showed that spore biomass could be prepared both in liquid and on solid nutrient media. Methodically, spore biomass preparation in liquid nutrient medium is preferable. The tests showed that spores emerged from anabiosis non-uniformly and it depended on original seed spore storage period. Spore cultures stored less than one year emerged from anabiosis more quickly. It was found that the spores formed more readily when the cultures were aerated with oxygen as well as that lag-phase culture medium had a stimulating effect on Bacillus subtilis spore germination. Bacillus subtilis strains were found to have antagonistic effect on Escherichia coli, Salmonella and Staphylococcus. Area of growth inhibition of the said bacteria was 15–20 mm. Tested Bacillus subtilis strains could be proposed for use as probiotics.

Anthrax toxins and capsules, which are encoded by genes located on pXO1 and pXO2, respectively, are major virulence factors of Bacillus anthracis. Our previous studies demonstrated that exposure to high-temperatures is unable to abolish the pXO1 plasmid of the Pasteur II strain, but the growth of the strain was obviously slower than that of the Sterne strain and wild-type virulent strain. To elucidate a potential regulatory mechanism of slowing growth, we employed comparative genome and bioinformatic analysis and revealed a unique SNP (G to T) at the 143135 bp position in pXO1 that is possibly involved in the mediation of growth of Pasteur II. However, the T to G mutation in groR did not result in any change of the amino acid sequence. A predominant nucleotide G existed at the 143135 bp in pXO1 of 100 wild-type B. anthracis isolates and 9 isolates documented in GenBank, whereas T replaced G in pXO1 of the Pasteur II strain. Further analysis indicate that the SNP is located in a gene between 143042 and 143173 bp, and that it encodes a small protein of 43 amino acids and is termed as a growth regulator (GroR). Site-directed mutagenesis and gene deletion demonstrates that groR regulates the growth and spore formation of B. anthracis. Our results indicate that the pXO1 plasmid is involved in the regulation of growth and spore formation in B. anthracis.