Introgressive hybridization is the main method of broadening the genetic diversity of bread wheat. Wild barley Hordeum marinum ssp. gussoneanum Hudson (2n = 4x = 28) has useful agronomical traits, such as high resistance to stress factors, that could be a potential source of new genes for bread wheat improvement. This study aimed to evaluate the possibility of introgression of H. marinum chromosomes into the genome of bread wheat using an incomplete amphiploid H. marinum ssp. gussoneanum (4x)–T. aestivum (Pyrotrix 28) (2n = 54) carrying the cytoplasm of wild barley. For this purpose, we crossed the line of bread wheat variety Pyrotrix 28 with an incomplete amphiploid, and then selected cytogenetically stable 42­chromosome plants with a high level of fertility in hybrid progeny. Genomic in situ hybridization (GISH) revealed a pair of H. marinum chromosomes in the genome of these plants. C­ banding analysis confirmed that bread wheat chromosome 4B was replaced by wild barley chromosome 4H^mar. SSR markers Xgwm368 and Xgwm6 confirmed the absence of chromosome 4B, and EST markers BAWU808 and BAW112 identified chromosome 4Hmar in the genome of the isolated disomic wheat­barley substitution line. The study of this line showed that the substitution of chromosome 4B with chromosome 4H^mar resulted in a change of some morphological traits. It included intense anthocyanin coleoptile coloration, specific for H. marinum, as well as a lack of purple coloration of the ears in the leaf sheath, specific for Pyrotrix 28. Line 4Hmar(4B) showed increased performance for several traits, including plant height, number of spikes and tillers per plant, spikelet and grain number in the main spike, grain number per plant, but it had decreased values of 1000­grain weight compared to wheat. Cytogenetic stability and fertility of line 4H^mar(4B) indicated a high compensation ability of barley 4H^mar for wheat chromosome 4B and confirmed their homeology.

Wild and domesticated emmer (ВВАА, 2n = 28) are of significant interest for expanding the genetic diversity of common wheat as sources of a high protein and microelement grain content, resistance to many biotic and abiotic factors. Particular interest in these species is also determined by their close relationship with Triticum aestivum L., which facilitates interspecific hybridization. The objective of this work was to analyze the nature of alien introgressions in hybrid lines from crossing common wheat varieties with T. dicoccoides and T. dicoccum, and to assess the effect of their genome fragments on the cytological stability of introgression lines. A C-banding technique and genotyping with SNP and SSR markers were used to determine localization and length of introgression fragments. Assessment of cytological stability was carried out on the basis of chromosome behavior in microsporogenesis. A molecular cytogenetic analysis of introgression wheat lines indicated that the inclusion of the genetic material of wild and domesticated emmer was carried out mainly in the form of whole arms or large fragments in the chromosomes of the B genome and less extended inserts in the A genome. At the same time, the highest frequency of introgressions of the emmer genome was observed in chromosomes 1A, 1B, 2B, and 3B. The analysis of the final stage of meiosis showed a high level of cytological stability in the vast majority of introgression wheat lines (meiotic index was 83.0–99.0 %), which ensures the formation of functional gametes in an amount sufficient for successful reproduction. These lines are of interest for the selection of promising material with agronomically valuable traits and their subsequent inclusion in the breeding process.

Charcoal rot (CR) caused by the fungal pathogen Macrophomina phaseolina is a devastating disease affecting soybean (Glycine max (L.) Merrill.) worldwide. Identifying the genetic factors associated with resistance to charcoal rot is crucial for developing disease-resistant soybean cultivars. In this research, we conducted a genome-wide association study (GWAS) using different models and genotypic data to unravel the genetic determinants underlying soybean resistance to сharcoal rot. The study relied on a panel of 252 soybean accessions, comprising commercial cultivars and breeding lines, to capture genetic variations associated with resistance. The phenotypic evaluation was performed under natural conditions during the 2021–2022 period. Disease severity and survival rates were recorded to quantify the resistance levels in the accessions. Genotypic data consisted of two sets: the results of genotyping using the Illumina iSelect 6K SNP (single-nucleotide polymorphism) array and the results of whole-genome resequencing. The GWAS was conducted using four different models (MLM, MLMM, FarmCPU, and BLINK) based on the GAPIT platform. As a result, SNP markers of 11 quantitative trait loci associated with CR resistance were identified. Candidate genes within the identified genomic regions were explored for their functional annotations and potential roles in plant defense responses. The findings from this study may further contribute to the development of molecular breeding strategies for enhancing CR resistance in soybean cultivars. Marker-assisted selection can be efficiently employed to accelerate the breeding process, enabling the development of cultivars with improved resistance to сharcoal rot. Ultimately, deploying resistant cultivars may significantly reduce yield losses and enhance the sustainability of soybean production, benefiting farmers and ensuring a stable supply of this valuable crop.

Apple clonal rootstocks are the basis of modern intensive horticulture, providing a rapid increase in yield and convenience of fruit trees cultivation. Production of clonal rootstocks under high humidity often causes powdery mildew infection caused by the pathogenic fungus Podosphaera leucotricha Salm., which significantly reduces the productivity of stoolbed. Growing powdery mildew resistant genotypes is the most appropriate way to combat this disease and allows reducing the use of fungicides. To accelerate the search for resistant forms, molecular markers associated with resistance genes have been developed. However, these markers have not been used to study clonal rootstocks. The aims of the work were the field assessment of powdery mildew resistance of apple clonal rootstocks from the collection of the Michurinsk State Agrarian University and the screening of the collection for Pl-1, Pl-2, Pl-w and Pl-d resistance genes. The results of a three-year field evaluation of powdery mildew resistance of 80 rootstocks allowed us to distinguish five main groups ranging from very low to highly resistant. A group of 57 accessions was classified as powdery mildew resistant. The search for resistance genes was performed using the AT20 SCAR (Pl-1 gene), OPU02 SCAR (Pl- 2 gene), EM DM01 (Pl-d gene), and EM M02 (Pl-w gene) markers. The Pl-d and Pl-1 genes identified in 33 (41.25 %) and 31 (38.75 %) accessions, respectively, were the most common in the collection. The Pl-w gene was detected only in two accessions. Identification of the Pl-2 gene with the OPU02 SCAR marker did not reveal a fragment of the expected size. Thirty accessions with different powdery mildew resistance scores had two genes, Pl-1 and Pl-d, and highly resistant forms G16 and 14-1 had a combination of the Pl-d and Pl-w genes. These accessions can be used as donors of powdery mildew resistance for breeding new apple clonal rootstocks.

Septoria is one of the harmful diseases of wheat cultivars cultivated in the Saratov region. This infectious disease of fungal etiology limits yield indicators and rapidly progresses in many regions of the Russian Federation. The aim of the research was to assess the resistance of winter and spring wheat cultivars that are referred to as promising and recommended for cultivation in the Low Volga region of the Russian Federation to pathogens of Septoria, to study the populations of Parastagonospora nodorum and P. pseudonodorum in the territory of the Saratov region in order to detect the presence of effector genes. Using molecular markers, we performed the identification of genes encoding NEs in 220 Parastagonospora spp. fungal isolates obtained from 7 cultivars of soft winter wheat, 6 taken from the winter triticale, 5 from soft spring wheat, 3 from durum spring wheat and 1 from spring oats. Among the P. nodorum isolates studied, there were both single genes Tox1, Tox3, and ToxA, and combinations of two genes in one genotype. The presence of the ToxA gene was not noted in the genotype of P. pseudonodorum isolates. During 2020–2022, a collection of winter and spring wheat cultivars was studied to detect resistance to Septoria blotch in field conditions (13 cultivars of winter wheat and 7 cultivars of spring wheat accordingly). The resistance of the cultivars was proven by laboratory evaluation. Three inoculums were used, including the isolates of Z. tritici, P. nodorum (ToxA, Tox1, Tox3), P. pseudonodorum (ToxA, Tox1, Tox3) mainly obtained from Saratov populations of 2022 (except for P. pseudonodorum with the ToxA gene). The tested cultivars were characterized using the Xfcp623 molecular marker, diagnostic for Tsn1/ tsn1 genes, which controls sensitivity to the fungal toxin of PtrToxA. Of greatest interest are 11 wheat genotypes that showed resistance to one, two and three species which served as causative agents of Septoria blotch (Zymoseptoria tritici, P. nodorum, P. pseudonodorum). These are the soft winter wheat cultivars Gostianum 237 (tsn1), Lutescens 230 (Tsn1), Guberniya (Tsn1), Podruga (Tsn1), Anastasia (Tsn1), Sosedka (Tsn1) and the soft spring wheat cultivars Favorit (tsn1), Prokhorovka (tsn1), Saratovskaya 70 (tsn1), Saratovskaya 73 (tsn1), Belyanka (tsn1). The results obtained are of interest as they might increase the efficiency of selection based on the elimination of genotypes with dominant Tsn1 alleles sensitive to PtrToxA. In addition to the economic value of the cultivars studied, it is recommended to use them in breeding for resistance to Septoria blotch.

The article outlines a brief historical background on the introduction to cultivation, distribution and breeding of spring durum wheat in the steppe and forest-steppe regions of Eurasia (the countries of the former USSR: Russia, Ukraine, and Kazakhstan). The approaches and methodology for improving durum wheat during certain scientific selection periods are given. The features of the selection program implementation and the breeding scale expansion during the creation of breeding stations at the beginning of the XX century, after the end of the Great Patriotic War, in the second half of the XX century, and at present are considered. A characteristic according to the main features and properties of varieties created in different periods is given. The achievements of the classical breeding method by comparing old and new varieties are analyzed. The efficiency and rate of wheat selection by periods in different regions of Russia is estimated. The results and methods of breeding for yield, resistance to drought, leaf diseases (Stagonospora nodorum Berk., Septoria tritici (Roeb. et Desm.), Bipolaris sorokiniana (Sacc.) Shoemaker, Pyrenophora tritici repentis (Died.) Drechs., Fusarium sp., Puccinia titicina Eriks., Puccinia graminis Pers. f. sp. tritici Eriks., Blumeria graminis (DC.) f. sp. tritici Em. Marchal), grain pathogens Ustilago tritici (Pers.) Rostr.) and pathogens causing darkening of the corcule and endosperm (Bipolaris sorokiniana (Sacc.) Shoemaker, Alternaria tenuis (Nees et Fr.), Аlternaria triticina (Prasada & Prabhu)), pests (Cephus pygmeus Lens, Osinosoma frit L., Mayetiola destructor (Say)), grain quality (protein content, amount of yellow pigments, dough rheology, sprouting resistance) and end products are presented. The prospects for the molecular marker application for a number of traits in breeding in the near future are given.

Wheat (Triticum aestivum L.) is a staple food and major source of dietary calories in Pakistan. Improving wheat varieties with higher grain yield and disease resistance is a prime objective. The knowledge of genetic behaviour of germplasm is key. To achieve this objective, elite wheat varieties were crossed in 4 by 3, line × tester design, and tested in 2019 in a triplicate yield trial to estimate genetic variance, general and specific combining ability, mid-parent heterosis and stripe rust (Puccinia striiformis L.). High grain 3358 kg·ha^–1 was recorded in F₁ hybrid (ZRG-79 × PAK-13). Analysis of variance (ANOVA) revealed significant genotypic variance in grain yield. Broad sense heritability (H²) was recorded in the range of 28 to 100 %. General combining ability (GCA) significant for grain yield in parents except FSD-08 and PS-05 was recorded, while specific combining ability (SCA) was recorded to be highly significant for grain yield only in two crosses (ZRG-79 × NR-09 and ZRG-79 × PAK-13). Mid-parent heterosis was estimated in the range of –28 to 62.6 %. Cross combinations ZRG-79 × PAK-13 depicted highly significant mid-parent heterosis (62.6 %). Highly significant correlation was observed among spike length, spikelets per spike, plant height and 1000-grain weight. Rust resistance index was recorded in the range of 0 to 8.5. These findings suggest exploitation of GCA for higher grain yield is important due to the presence of additive gene action and selection in the filial generations will be effective with improved rust resistance, while cross combinations ZRG-79 × PAK-13 high GCA are best suited for hybrid development.

Triticum timopheevii Zhuk. attracts the attention of bread wheat breeders with its high immunity to the leaf rust pathogen. However, introgressions from this species in Triticum aestivum L. are little used in practical breeding. In the presented study, the agronomic value of T. aestivum/T. timopheevii line L624 was studied in comparison with the parent cultivars Saratovskaya 68, Dobrynya and the standard cultivar Favorit during 2017–2022. Introgressions from T. timopheevii in L624 were detected by the FISH method with probes pSc119.2, pAs1 and Spelt1, as well as micro satellite markers Xgwm312, Xgpw4480 and Xksum73. Translocations of 2AS.2AL-2A^tL and on 2DL were detected as well. Line L624 is highly resistant to Puccinia triticina both under the background of natural epiphytotics and under laboratory conditions. PCR analysis with the DNA marker of the LrTt1 gene (Xgwm312) revealed that it is not identical to the Lr gene(s) in L624. According to a five-year study, the grain yield of L624 was, on average, higher than that of Favorit and Dobrynya, but lower than that of Saratovskaya 68. Line L624 had a lower weight of 1000 grains than the recipients, and was at the same level with the standard cultivar Favorit. Introgressions from T. timopheevii in L624 increased the grain protein content by comparison with Saratovskaya 68 and Favorit, but it was at the same level as in Dobrynya. As for parameters of flour and bread, L624 was not inferior to the recipient cultivars, but by volume and porosity of bread, it surpassed Saratovskaya 68. Moreover, L624 surpassed Favorit by the elasticity of the dough, the ratio of the elas ticity of the dough to the extensibility and the strength of the flour. Thus, the results obtained suggest that introgressions in chromosomes 2A and 2D in L624 do not impair baking properties.

Orthotopic transplantation of glioblastoma cells in the brain of laboratory mice is a common animal model for studying brain tumors. It was shown that 1H magnetic resonance spectroscopy (MRS) enables monitoring of the tumor’s occurrence and its development during therapy based on the ratio of several metabolites. However, in studying new approaches to the therapy of glioblastoma in the model of orthotopic xenotransplantation of glioma cells into the brain of mice, it is necessary to understand which metabolites are produced by a growing tumor and which are the result of tumor cells injection along the modeling of the pathology. Currently, there are no data on the dynamic metabolic processes in the brain that occur after the introduction of glioblastoma cells into the brain of mice. In addition, there is a lack of data on the delayed effects of invasive brain damage. Therefore, this study investigates the long-term dyna mics of the neurometabolic profile, assessed using ¹H MRS, after intracranial injection of a culture medium used in orthotopic modeling of glioma in mice. Levels of N-acetylaspartate, N-acetylaspartylglutamic acid, myoinositol, taurine, glutathione, the sum of glycerophosphocholine and phosphocholine, glutamic acid (Glu), glutamine (Gln), and gamma aminobutyric acid (GABA) indicate patterns of neurometabolites in the early stage after intracranial injection similar to brain trauma ones. Most of the metabolites, with the exception of Gln, Glu and GABA, returned to their original values on day 28 after injection. A progressive increase in the Glu/Gln and Glu/GABA ratio up to 28 days after surgery potentially indicates an impaired turnover of these metabolites or increased neurotransmission. Thus, the data indicate that the recovery processes are largely completed on day 28 after the traumatic event in the brain tissue, leaving open the question of the neurotransmitter system impairment. Consequently, when using animal models of human glioma, researchers should clearly distinguish between which changes in neurometabolites are a response to the injection of cancer cells into the brain, and which processes may indicate the early development of a brain tumor. It is important to keep this in mind when modeling human glioblastoma in mice and monitoring new treatments. In addition, these results may be important in the development of approaches for non-invasive diagnostics of traumatic brain injury as well as recovery and rehabilitation processes of patients after certain brain surgeries.

Germline-restricted chromosomes (GRCs) are present in the genomes of germline cells and absent from somatic cells. A GRC is found in all species of the songbirds (Passeri) and in none of the other bird orders studied to date. This indicates that GRC originated in the common ancestor of the songbirds. The germline-restricted chromosome is permanently absent from somatic cells of the songbird, while female germline cells usually contain two copies of GRC and male ones have one copy. In females, GRCs undergo synapsis and restricted recombination in their terminal regions during meiotic prophase. In males, it is almost always eliminated from spermatocytes. Thus, GRC is inherited almost exclusively through the maternal lineage. The germline-restricted chromosome is a necessary genomic element in the germline cells of songbirds. To date, the GRC genetic composition has been studied in four species only. Some GRC genes are actively expressed in female and male gonads, controlling the development of germline cells and synthesis of the proteins involved in the organization of meiotic chromosomes. Songbird species vary in GRC size and genetic composition. The GRC of each bird species consists of amplified and modified copies of genes from the basic genome of that species. The level of homology between GRCs of different species is relatively low, indicating a high rate of genetic evolution of this chromosome. Transmission through the maternal lineage and suppression of the recombination contribute significantly to the accelerated evolution of GRCs. One may suggest that the rapid coordinated evolution between the GRC genes and the genes of the basic genome in the songbirds might be responsible for the explosive speciation and adaptive radiation of this most species-rich and diverse infraorder of birds.

The hippocampus plays the key role in stress response regulation, and stress response appears to be weakened in domesticated animals compared to their wild relatives. The hippocampus is functionally heterogeneous along its dorsoventral axis, with its ventral compartment being more closely involved in stress regulation. An earlier series of experiments was conducted with a unique breeding model of animal domestication, the farm silver fox (Vulpes vulpes), which included tame, aggressive, and unselected animals. A decrease in many indices of the hypothalamic–pituitary–adrenal activity was observed in tame animals. Also, adult hippocampal neurogenesis was more intense in tame foxes, and this fact may relate to reduced stress levels in this experimental population of foxes. Nevertheless, the molecular mechanisms responsible for the reduced stress response in tame animals remain obscure. In this study, serum cortisol levels and the mRNA levels of 13 genes in the dorsal and ventral hippocampus have been measured and compared in tame, aggressive, and unselected foxes. At the current stage of domestication, stress-induced cortisol levels in tame, aggressive, and unselected animals differ significantly from each other: tame foxes show the lowest levels, and aggressive ones, the highest. Twelve genes tested demonstrate significant gene expression differences between the dorsal and ventral hippocampi. These differences are mainly consistent with those found in rodents and humans. In tame foxes, significantly elevated mRNA levels were recorded for several genes: CYP26B1 for cytochrome P450 26B1 and ADRA1A for α_1A adrenergic receptor in the dorsal hippocampus, whereas the level of NR3C2 mRNA for mineralocorticoid receptor was higher in the ventral. It is presumed that these genes constitute an important part of the mechanism reducing stress induced by contacts with humans and contribute to linking stress regulation with adult neurogenesis in tame foxes and domesticated animals in general.

Single nucleotide polymorphisms (SNPs) are the most common type of variation in the human genome. The vast majority of SNPs identified in the human genome do not have any effect on the phenotype; however, some can lead to changes in the function of a gene or the level of its expression. Most SNPs associated with certain traits or pathologies are mapped to regulatory regions of the genome and affect gene expression by changing transcription factor binding sites. In recent decades, substantial effort has been invested in searching for such regulatory SNPs (rSNPs) and understanding the mechanisms by which they lead to phenotypic differences, primarily to individual differences in susceptibility to di seases and in sensitivity to drugs. The development of the NGS (next-generation sequencing) technology has contributed not only to the identification of a huge number of SNPs and to the search for their association (genome-wide association studies, GWASs) with certain diseases or phenotypic manifestations, but also to the development of more productive approaches to their functional annotation. It should be noted that the presence of an association does not allow one to identify a functional, truly disease-associated DNA sequence variant among multiple marker SNPs that are detected due to linkage disequilibrium. Moreover, determination of associations of genetic variants with a disease does not provide information about the functionality of these variants, which is necessary to elucidate the molecular mechanisms of the development of pathology and to design effective methods for its treatment and prevention. In this regard, the functional analysis of SNPs annotated in the GWAS catalog, both at the genome-wide level and at the level of individual SNPs, became especially relevant in recent years. A genome-wide search for potential rSNPs is possible without any prior knowledge of their association with a trait. Thus, mapping expression quantitative trait loci (eQTLs) makes it possible to identify an SNP for which – among transcriptomes of homozygotes and heterozygotes for its various alleles – there are differences in the expression level of certain genes, which can be located at various distances from the SNP. To predict rSNPs, approaches based on searches for allele-specific events in RNA-seq, ChIP-seq, DNase-seq, ATAC-seq, MPRA, and other data are also used. Nonetheless, for a more complete functional annotation of such rSNPs, it is necessary to establish their association with a trait, in particular, with a predisposition to a certain pathology or sensitivity to drugs. Thus, approaches to finding SNPs important for the development of a trait can be categorized into two groups: (1) starting from data on an association of SNPs with a certain trait, (2) starting from the determination of allele-specific changes at the molecular level (in a transcriptome or regulome). Only comprehensive use of strategically different approaches can considerably enrich our knowledge about the role of genetic determinants in the molecular mechanisms of trait formation, including predisposition to multifactorial diseases.

Ectodermal dysplasia (ED) is a heterogeneous group of hereditary diseases of the skin and its appendages, which are characterized by impaired development and/or homeostasis of two or more ectoderm derivatives, including: hair, teeth, nails, sweat glands and their modifications (mammary glands, for instance). The overall prevalence of ectodermal dysplasia remains precisely unknown not only in Russia, but also in the world, nor is known the contribution of individual genes to its structure. This complicates the DNA diagnosis establishment of this disease due to the lack of an accurate diagnostic algorithm and a universal cost-effective method of analysis. To date, the most highly-researched genes involved in the development of anhydrous or hypohidrotic forms of ED are EDA, EDAR, EDARADD and WNT10A. The ectodysplasin A (EDA) gene is the cause of the most common X-linked form of ED, a gene from the Wnt family (WNT10A) is responsible for the autosomal recessive form of the disease, and two other genes (EDAR and EDARADD) can cause both autosomal recessive and autosomal dominant forms. This review provides the characteristics of the genes involved in ED, their mutation spectra, the level of their expression in human tissues, as well as the interrelation of the aforementioned genes. The domain structures of the corresponding proteins are considered, as well as the molecular genetic pathways in which they are involved. Animal models for studying this disorder are also taken into consideration. Due to the cross-species genes conservation, their mutations cause the disruption of the development of ectoderm derivatives not only in humans, but also in mice, cows, dogs, and even fish. It can be exploited for a better understanding of the etiopathogenesis of ectodermal dysplasias. Moreover, this article brings up the possibility of recurrent mutations in the EDA and WNT10A genes. The review also presents data on promising approaches for intrauterine ED treatment.

The review describes the main methods for assessing directional selection in human populations. These include bioinformatic analysis of DNA sequences via detection of linkage disequilibrium and of deviations from the random distribution of frequencies of genetic variants, demographic and anthropometric studies based on a search for a correlation between fertility and phenotypic traits, genome-wide association studies on fertility along with genetic loci and polygenic risk scores, and a comparison of allele frequencies between generations (in modern samples and in those obtained from burials). Each approach has its limitations and is applicable to different periods in the evolution of Homo sapiens. The main source of error in such studies is thought to be sample stratification, the small number of studies on nonwhite populations, the impossibility of a complete comparison of the associations found and functionally significant causative variants, and the difficulty with taking into account all nongenetic determinants of fertility in contemporary populations. The results obtained by various methods indicate that the direction of human adaptation to new food products has not changed during evolution since the Neolithic; many variants of immunity genes associated with inflammatory and autoimmune diseases in modern populations have undergone positive selection over the past 2–3 thousand years owing to the spread of bacterial and viral infections. For some genetic variants and polygenic traits, an alteration of the direction of natural selection in Europe has been documented, e. g., for those associated with an immune response and cognitive abilities. Examination of the correlation between fertility and educational attainment yields conflicting results. In modern populations, to a greater extent than previously, there is selection for variants of genes responsible for social adaptation and behavioral phenotypes. In particular, several articles have shown a positive correlation of fertility with polygenic risk scores of attention deficit/hyperactivity disorder.

The diversity of macroinvertebrates, the structure of their communities in Bolshiye Koty Bay (Lake Baikal) was studied by a DNA metabarcoding approach using an Illumina MiSeq system. Internal primer mlCOIintF in combination with jgHCO2198 of the Folmer fragment of the COI gene were used for macroinvertebrate metabarcoding. A total of 118009 reads of the COI gene fragment (at least 313 bp in length) were obtained. The correlation of the Spearman coefficient (S = 0.6, p < 0.05) with the abundance of macroinvertebrates in the samples before DNA extraction showed that the number of reads can serve as an indirect characteristic of the abundance of a species (operational taxonomic unit, OTU). 115 OTUs belonging to the higher taxa of macroinvertebrates were identified: Porifera, 1; Platyhelminthes, 3; Annelida, 38; Arthropoda, 55; Mollusca, 18. At a high level of resolution (with homology with GenBank reference sequences ≥ 95 %, coverage ≥ 90 %), 46 taxa of macroinvertebrates comprising three communities were registered: one dominated by molluscs (Choanomphalus conf. maacki) and two dominated by chironomids (Orthocladius grega rius Linev., Sergentia baicalensis Tshern.). Communities are characterized by low species diversity according to Shannon (from 0.7 to 1.2 bits), high concentration of dominance according to Simpson (from 0.5 to 0.7) and low evenness according to Pielou (from 0.3 to 0.4). Dominants and subdominants in the communities account for 91 to 96 % of COI gene fragment reads. The spatial distribution of the dominant species identified in the communities is influenced by the geomorphological features of the bottom and the composition of sediments in the area studied. The approach proposed for studying the structure of macroinvertebrate communities based on DNA metabarcoding and next generation sequencing can be recommended for express assessment of the state of aquatic ecosystems in the monitoring.

Microeukaryotes are vital for maintaining soil quality and ecosystem functioning, however, their communities are less studied than bacterial and fungal ones, especially by high throughput sequencing techniques. Alveolates are important members of soil microbial communities, being consumers and/or prey for other microorganisms. We studied alveolate diversity in soil under the undisturbed steppe (US) and cropped for wheat using two tillage practices (conventional, CT, and no-till, NT) by amplifying the ITS2 marker with ITS3_KYO2/ITS4 primers and sequencing amplicons using Illumina MiSeq. A total of 198 Alveolata OTUs were identified, with 158 OTUs attributed to the Ciliophora phylum, containing five classes: Litostomatea, Spirotrichea and Oligohymenophorea, Nassophorea and Phyllopharyngea. Litostomatea and Phyllopharyngea were more abundant in US as compared with CT and NT. The observed OTU richness was higher in US than in CT and NT. The β-biodiversity of soil ciliates also very distinctly differentiated the US field from CT and NT. In the US, Nassophorea and Spirotrichea correlated positively with sand and negatively with clay, silt and SOM contents. This is the first report about soil ciliates diversity in Siberia as assessed by metabarcoding technique. The revealed clear effect of land use on the relative abundance of some taxa and a lack of tillage effect suggest the importance of the quantity and quality of plant material input for shaping the prey for ciliates. The ITS-metabarcoding technique was used for the first time in the research of ciliates diversity; further studies, embracing diverse aspects of soil ciliates by combining -omics methodology with the traditional one, are needed to get a better insight on the ecological roles of the main ciliate taxa in the complex soil system.

The monkeypox epidemic, which became unusually widespread among humans in 2022, has brought awareness about the necessity of smallpox vaccination of patients in the risk groups. The modern smallpox vaccine variants are introduced either intramuscularly or by skin scarification. Intramuscular vaccination cannot elicit an active immune response, since tissues at the vaccination site are immunologically poor. Skin has evolved into an immunologically important organ in mammals; therefore, intradermal delivery of a vaccine can ensure reliable protective immunity. Historically, vaccine inoculation into scarified skin (the s.s. route) was the first immunization method. However, it does not allow accurate vaccine dosing, and high-dose vaccines need to be used to successfully complete this procedure. Intradermal (i.d.) vaccine injection, especially low-dose one, can be an alternative to the s.s. route. This study aimed to compare the s.s. and i.d. smallpox immunization routes in a mouse model when using prototypic second- and fourth-generation low-dose vaccines (104 pfu). Experiments were conducted using BALB/c mice; the LIVP or LIVP-GFP strains of the vaccinia virus (VACV) were administered into the tail skin via the s.s. or i.d. routes. After vaccination (7, 14, 21, 28, 42, and 56 days post inoculation (dpi)), blood samples were collected from the retro-orbital venous sinus; titers of VACV-specific IgM and IgG in the resulting sera were determined by ELISA. Both VACV strains caused more profound antibody production when injected via the i.d. route compared to s.s. inoculation. In order to assess the level of the elicited protective immunity, mice were intranasally infected with a highly lethal dose of the cowpox virus on 62 dpi. The results demonstrated that i.d. injection ensures a stronger protective immunity in mice compared to s.s. inoculation for both VACV variants.