[The Proteome of Extracellular Membrane Vesicles from Bacillus pumilus 3-19].

Production of extracellular membrane vesicles plays an important role in communication in bacterial populations and in bacteria-host interactions. Vesicles as carriers of various regulatory and signaling molecules may be potentially used as disease biomarkers and promising therapeutic agents, including vaccine preparations. The composition of membrane vesicles has been deciphered for a limited number of Gram-negative and Gram-positive bacteria. In this work, for the first time, extracellular membrane vesicles of a streptomycin-resistant strain Bacillus pumilus 3-19, a producer of extracellular guanyl-preferring ribonuclease binase, are isolated, visualized, and characterized by their genome and proteome composition. It has been established that there is no genetic material in the vesicles and the spectrum of the proteins differs depending on the phosphate content in the culture medium of the strain. Vesicles from a phosphate-deficient medium carry 49 unique proteins in comparison with 101 from a medium with the high phosphate content. The two types of vesicles had 140 mutual proteins. Flagellar proteins, RNase J, which is the main enzyme of RNA degradosomes, phosphatases, peptidases, iron transporters, signal peptides, were identified in vesicles. Antibiotic resistance proteins and amyloid-like proteins whose genes are present in B. pumilus 3-19 cells are absent. Phosphate deficiency-induced binase was found only in vesicles from a phosphate-deficient medium.

Severe decline in abundance of Cyathostoma lari, a parasite of the nasal and orbital sinuses of gulls, at their central European nesting grounds.

Cyathostoma lari is a parasite of the nasal and orbital sinuses of gulls and other hosts in Europe and Canada. Here, we provide an overview of previously published data on the prevalence and infection intensity of C. lari in gulls. Furthermore, based on our data, we analyze the spatiotemporal trends in the prevalence and intensity of infection by C. lari in Chroicocephalus ridibundus in Czechia (central Europe; data from 1964 to 2014) and compare them with those obtained from five species of gulls in Karelia (Northwest Russia; data from 2012-2020). Based on our preliminary observations, we hypothesized that C. lari is subject to a decline in certain regions, but this decline is not necessarily applicable throughout its distribution range. We found that the C. lari population crashed in specific parts of its distribution range. The reasons are unknown, but the observed population changes correspond with the diet switch of their core host in Czechia, C. ridibundus. We previously observed a diet switch in Czech C. ridibundus from earthworms (intermediate hosts of C. lari) to other types of food. This diet switch affected both young and adult birds. Nevertheless, it may not necessarily affect populations in other regions, where they depend less on earthworms collected from agrocenoses affected by agrochemicals and trampling. Correspondingly, we found that these changes were limited only to regions where the gulls feed (or fed) on arable fields. In Karelia, where arable fields are scarce, gulls likely continue to feed on earthworms and still display high infection rates by C. lari. Therefore, C. lari, a parasite of the nasal and orbital sinuses of gulls, nearly disappeared from their central European nesting grounds but is still present in better-preserved parts of its distribution range.

[Microbiome of therapeutic muds used in Tatarstan]. / Mikrobiom lechebnykh gryazei, primenyaemykh v Tatarstane.

Therapeutic muds (peloids), which are widely used for body healing, improve metabolism and have antibacterial, anti-inflammatory and analgesic effects due to enrichment with necessary microelements and biological active substances. However, the microbiological component of these effects is not well studied. OBJECTIVE: To characterize the microbiome of therapeutic muds, used in the Tatarstan Republic, by identifying spectrum of cultivated microorganisms, using molecular analysis of bacterial communities, and by determining their biodiversity and functional potential based on revealed genetic determinants. MATERIAL AND METHODS: The study design of 5 peloids samples (local sapropels and peat deposits of swamp; 3 samples of Crimean sulfide muds) included three main techniques: isolation and taxonomic determination of cultivated microorganisms by time-of-flight mass-spectrometry; molecular analysis of peloids bacterial communities by 16S RNA high-throughput sequencing; identification of functional profiles of communities by their genetic determinant using Global Mapper tool on iVikodak platform. RESULTS: Experimental studies have confirmed the safety of examined peloids, where non-pathogenic cultivated bacteria belonging mainly to Bacillus and Rhodococcus genera were dominant. Metagenomic analysis showed that Firmicutes, Proteobacteria and Actinobacteria were predominant in all samples in different ratios. It has been established, that there is both the internal biodiversity of each sample and difference between them. The functional profile of microbial communities was determined based on the identification of bacterial genes. It has been revealed that all communities have an ability to synthesize antibiotics, as well as to decompose dangerous xenobiotics - polyaromatic hydrocarbons, cyclic compounds, and dioxins. CONCLUSION: Various microbial communities, which were identified in the therapeutic muds, contribute both to the clearance of toxicants in the peloids and to the antibacterial properties of the latter. The obtained priority results create a fundamental basis for the subsequent study of the role of peloids' microbiome of different origin in their healing action.

No strict host specificity: Brain metacercariae Diplostomum petromyzifluviatilis Müller (Diesing, 1850) are conspecific with Diplostomum sp. Lineage 4 of Blasco-Costa et al. (2014).

Metacercariae of Diplostomum petromyzifluviatilis (Digenea, Diplostomidae) from the brain of European river lamprey Lampetra fluviatilis from the Baltic Sea basin and Arctic lamprey Lethenteron camtschaticum from the White Sea basin were studied with the use of genetic and morphological methods. Phylogenetic analysis based on cox1 marker showed that the parasites of both lamprey species were conspecific with Diplostomum sp. Lineage 4 of Blasco-Costa et al. (2014). The name Diplostomum petromyzifluviatilis Müller (Diesing, 1850) has historical precedence as a species described from the brain of lampreys and should be used in genus nomenclature. There were no morphological qualitative differences between the metacercariae from the two lamprey species but those from L. fluviatilis were larger than those from L. camtschaticum. We expanded the data on the second intermediate hosts and the localization of D. petromyzifluviatilis, showing that its metacercariae occur not only in the brain of lampreys but also in the brain and the retina of three-spined stickleback Gasterosteus aculeatus and the vitreous humour of the perch Perca fluviatilis across the European part of the Palearctic.

The Effect of Silver and Copper Nanoparticles on the Wheat-Pseudocercosporella herpotrichoides Pathosystem.

The paper covers the study of the effects of silver (Ag) and copper (Cu) nanoparticles on wheat-Pseudocercosporella herpotrichoides pathosystem in general and, separately, on their interaction both with the plant and with the pathogen. Plants, treated with nonionic colloidal solutions of biogenic metal nanoparticles of Ag and Cu, have taken seed treatment as stress and have demonstrated the same changes in the dynamic patterns of thiobarbituric acid reactive substances (TBARS) content as a seedling infection or in its combination with a nanoparticle treatment. The wheat variety, which is sensitive to pathogen action, has showed a substantial (100%) increase in the TBARS contents, while the other varieties has shown lesser (40%) changes in the TBARS content as compared to the control. Besides, both silver and copper nanoparticles have not affected the growth and development of P. herpotrichoides, thus suggesting that the effect of nanoparticles is determined by the plant's responses to the pathogen rather than the phytotoxic action of the copper or silver nanoparticles, at least during the initial stages of the pathological process.

[PARASITES OF THE COMMON ROACH (RUTILUS RUTILUS L.) UNDER THE IMPACT OF INDUSTRIAL POLLUTION OF A LAKE].

The consequences of man-induced transformation of Lake Kostomukshskoe (tailings dump) related to increased mineralization and entry of highly dispersed dredge material not typical for northern lakes were studied using roach parasites as the example. It was found that the roach parasite fauna has been losing rare and scant parasite species of different taxonomic groups, mainly helminthes with an indirect life cycle. Common and dominant species of myxosporidians, monogeneans, trematodes of genera Diplostomum and Tylodelphys have been preserved. The chances of survival are higher for those parasites with direct life cycle. Also the parasite species whose larvae can actively penetrate the host have been survived. The species Ligula intestinalis, Philometra rishta, Pseudocapillaria tomentosa had reported very rare. Fish are infected with these parasites by feeding of Copepoda and oligochaetes.

[RELATIONSHIP BETWEEN THE PREVALENCE OF CHRONIC NONINFECTIOUS DISEASES AND ELECTROPHYSICAL STATE OF THE ENVIRONMENT].

In the paper there is evaluated the relationship of features of electronic state of the environment with a level of chronic, noninfectious diseases (CNID) in the regions of Russia, obtained on the basis of the monitoring measurements of the intensity of natural background electronic Bose condensate (BEBC) of natural ecosystems in a number of Russian regions and seas of the Arctic Ocean. The assessment of BEBC was implemented on results of measurements of redox state of distilled water being in the contact with natural water. The equilibrium redox state of distilled water, determined by the influx of electrons (quantum reduction) outside, is proportional to the intensity of BEBC. The obtained data attest to an increase in the intensity of the background of EBC in Siberia regions and, especially, within the limits ofwaters of Lake Baikal (the redox potential of the surface water in the lake ~ -70mV). Also there is observed a strong dependence of the background EBC in the latitudinal direction. Low levels of background EBC were noted in the Arkhangelsk region and the north-eastern Chukotka. Functioning of international systems of plasma sounding of ionosphere (such systems as HAARP) were established to have a detrimental effect on the background EBC in these regions. According to the results of measurements of the relative values of intensities of natural background of Bose condensate of electrons there was constructed the dependence reflecting the relationship of the prevalence of noninfectious diseases in the regions of Russia with the redox state of distilled water which can be characterized as a significant (regression coefficient R2 = 0.78). The relationship between noninfectious diseases (NID, %) with the intensity of the background of EBC (Ib rel. units) is estimated by the equation: NID [%] = 0.24Eh [mV]-25, where Eh ~ I/Ib. Numerical evaluations show that an increase in the biosphere redox potential of water by 90mV leads to an increase of the primary incidence by 20% (relatively to the average values for Russia). Analysis of results attests to the relationship of CNID with the electrophysical state of the environment that allows from different positions to arrive to true causes of their emergence, associated with changes in the electrophysical conditions of habitation and human activities that lead to the nascency of cellular metabolic disturbances.

[Electronic deficit as a possible health risk factor].

There is presented the analysis of medical-demographic situation in Russia, and the increase in population mortality is shown both to be associated with the degradation of the geosphere and alongside with other factors determined by the change in the electronic state of the environment. On the base of the interrelationship between the electronic saturation of the environment and an increase in population mortality and morbidity there is established a such risk factor for human health and life, which may currently become one out of significant, videlicet, the electronic deficit. In conditions of its appearance there are proposed options solving this problem by means of elaboration of the scientific rationale for the impact of the electronic deficit on the human organism and the creation of technologies providing environmental--medical safety of the population by virtue of the correction of the electronic state of the human habitat, food and drinking water and the implementation of the system for monitoring electronic abundance of the environment.

[Effects of quantum nonlocality in the water activation process].

The dynamic alterations of the magnetic flux density of the water volume, activated with structurally stressed calcium carbonate in micellar form have been investigated. The phase of the associated water was established to exhibit electrical and magnetic properties, recorded by in B&E meter in the frequency range of 5Hz - 2kHz. Alterations in water Eh (redox) potential and the magnetic flux density B testify to synchronous auto-oscillatory changes. This gives evidence of non-linearity of the relationship between auto-oscillatory processes excited in the water; and reflects the nonlocal in time the relationship between the states of water, manifesting in a change of water activity on the 1st and 2nd day in negative time. The mechanism of action of associated water phase is shown to be described by de Broglie concept of matter waves with taking into account delocalized in time states of phase of electron wave packet in accordance with the transactional interpretation of quantum physics.

[Ion-radical oxygen species--the main indicator reflecting of the electron-donating ability of water].

Experimental evidence of the electron-donor ability of drinking water with ion-molecular forms of active oxygen is reported The concentration limits of the content of peroxide ion-radicals (48 mkg /L) in the absence of molecular hydrogen peroxide in drinking water has been determined. The concentration of the peroxide ion-radical in drinking water has been proposed to be used as an index of the water biocatalytic activity.

Phosphate-independent expression of the carbon-phosphorus lyase activity of Escherichia coli.

On the basis of mutational analysis, the genes for phosphonate uptake and degradation in Escherichia coli were shown to be organized in a 10.9-kb operon of 14 genes (named phnC to phnP) and induced by phosphate (P(i)) starvation [Metcalf and Wanner (1993) J Bacteriol 175: 3430-3442]. The repression of phosphonate utilization by P(i) has hindered both the biochemical characterization of the carbon-phosphorus (C-P) lyase activity and the development of improved methods for phosphonate biodegradation in biotechnology. We have cloned the genes phnG to phnP (associated with C-P lyase activity) with the lac promoter to provide expression of C-P lyase in the presence of P(i). A number of strains lacking portions of the phn operon have been constructed. In vivo complementation of the strains, in which phnC to phnP (including both Pn transport and catalysis genes) or phnH to phnP (including only catalysis genes) was deleted, with plasmids carrying various fragments of the phn operon revealed that the expression of phnC-phnP gene products is essential to restore growth on minimal medium with phosphonate as the sole phosphorus source, while phnG-phnM gene products are required for C-P lyase activity as assessed by in vivo methane production from methylphosphonic acid. The minimum size of the DNA required for the whole-cell C-P lyase activity has been determined to be a 5.8-kb fragment, encompassing the phnG to phnM genes. Therefore, there is no requirement for the phn CDE-encoded phosphate transport system, suggesting that cleavage of the C-P bond may occur on the outer surface of the inner membrane of E. coli cells, releasing the carbon moiety into the periplasm. These data are in agreement with the observation that phosphonates cannot serve as the carbon source for E. coli growth.

Characterization of the methylenediphosphonate transport system in Arthrobacter sp. GLP-1 using the novel tritium-labelled derivative.

The novel tritium-labelled derivative of methylenediphosphonate (MDP) was used in uptake studies of Arthrobacter sp. GLP-1 capable of utilizing a wide range of organophosphonates as its sole source of phosphorus. The MDP uptake was greatly stimulated upon phosphate deprivation. The uptake obeys Michaelis-Menten kinetics with respective Km and Vmax values of 33 microM and 0.3 nmol.min-1.mg-1 fr.wt. Glyphosate and pyrophosphate were competitive inhibitors of MDP uptake. The effect of orthophosphate was more complex than a mere inhibition of MDP uptake since activation occurred at low concentrations. The uptake of MDP by Arthrobacter sp. strain GLP-1 appears to be mediated by a transport system different from the glyphosate uptake system operating in the same cells. The driving force for MDP uptake by Arthrobacter sp. GLP-1 may be a proton gradient across the cell membrane.