Long-term studies (1975–2023) in the Central Laboratory of the Ecology of the Wolf Population Group using original methods confirm that the family consists of parents and only young individuals (up to 1 year). A different social structure is typical of packs with wolf-dog hybrids. On the basis of publications about stray (feral) dogs and their hybrids with wolves, the author proposed to identify hybrid packs as wolf-dog (female wolf) and dog-wolf (female dog): the former inherit the life strategy and lifestyle of the wolf, and the latter inherit that of dogs; therefore, they live in packs (parcels) of different ages all year round and can unite more than 18 individuals in a pack. Analysis of the social, spatial, and behavioral niches (“subniches”) of feral dogs, the wolf, and its hybrids showed that they occupy different ecological niches. The author not only proposed a new approach to studying the fundamental and realized niches of other animal species but also demonstrated it using the example of studies of the width of the trophic niche of wolf and elk populations.

An important component of animal life is behavior. The article examines two forms of behavior: cognitive, genetically fixed, and intellectual, acquired as an adaptation to the environment. The scientific novelty of the study lies in the interpretation of these forms using the example of nest-building, territorial and foraging behavior of families in the wolf population group. The conservatism of the cognitive and the plasticity of the intellectual are shown, which together ensure high vitality and stability of wolf populations in the biogeocenosis. As a result of a comparative analysis, it was found that with increasing contacts between a wolf and a moose (Central Forest Biosphere Reserve), and a wolf and a deer (Yellowstone National Park and southwestern Montana), changes in behavior occurred in the prey. In moose, this manifested itself in increased alertness, which made it difficult for the wolf to approach the prey at a distance for a successful attack, and deer left feeding areas, but with an increased risk of attack by a predator, and preferred safer habitats with better visibility to detect wolves. These are the consequences of turning on the natural caution reflex. Increased alertness or vigilance was a barrier against the predator eating the prey. But the wolf cannot change the genetically fixed process of killing the victim by changing the hunting strategy: moving from a short-term pursuit in case of an unsuccessful attack to a long one.

Structural and functional vital activity of each animal species organically fits into the life of the biosphere. To understand their place and role, a new approach to study methods is needed. Based on many years of research (1975–2022) on the ecology and behavior of the wolf, an attempt was made to understand the features of the structural and functional hierarchy of the life strategy of this species. It has been shown with argumentation that he strategy of the Individual is to adapt to living in the environment; the couple’s strategy is to form a habitat and fit into the general structure of the population; the family’s strategy is to raise and educate wolf cubs; the population’s strategy is to preserve genetic diversity, established adaptations, and sustainability in the environment; the strategy of the species is to preserve the habitat; the strategy of the community (biocenosis) consists in participation in the conservation and formation of the biodiversity of certain species; and the strategy of the ecosystem (biogeocoenosis) consists in participation in structural, functional, and energy processes.

The organization of the living space of animal populations is diverse, but in the generalized variant it is characterized by two concepts – “territory” and “habitat area”. The first provides for the protection of the “territory” by single individuals, married couples, families, flocks using acoustic signals and olfactory labels to mark boundaries. The second does not provide for such strict habitat protection. It is generally recognized that wolf families are characterized by lifestyle according to the “territory” type. But long-term studies of the ecology and behavior of wolf families (1975–2022) in the Central Forest Biosphere Reserve do not confirm this opinion: reasoned doubts about the “territoriality” of wolves are presented. The nature of the change in the size of the “territories”, the features of the placement of labels, the absence of conflict situations suggest the organization of living space according to the type of “habitat”.

Исследовано распространение антибиотикоустойчивых микроорганизмов и генов резистентности к антибиотикам в очистных сооружениях г. Пущино. Обнаружены и охарактеризованы плазмиды резистентности к антибиотикам как возможные векторы для распространения генов устойчивости в окружающей среде. Продемонстрирована роль катаболических плазмид в биодеградации углеводородов нефти. Исследован горизонтальный перенос плазмид в лабораторных условиях и в открытой окружающей среде в процессе утилизации полициклических ароматических углеводородов. Исследованы ризосферные бактерии рода Pseudomonas, относящиеся к уникальной группе PGPR. Штаммы продуцируют широкий спектр биологически активных метаболитов, включая сидерофоры, феназины, 2,4-диацетилфлороглюцин, пиолютеорин, пирролнитрин, оомицин А, цианогенные гликозиды, синтезируют фитогормоны, включая индолил-3-уксусную кислоту (ИУК) и стимулируют корнеобразование у растений. На основе одного из таких штаммов P. chlororaphis (ранее P. aureofaciens) BS1393 разработан биопрепарат «Псевдобактерин-2», обладающий высокой эффективностью против целого ряда заболеваний растений. The dissemination of antibiotic-resistant microorganisms and antibiotic resistance genes in treatment facilities of Pushchino was studied. The antibiotic resistance plasmids were isolated and characterized as possible vectors for the spread of resistance genes in the environment. The role of catabolic plasmids in the biodegradation of oil hydrocarbons was demonstrated. The horizontal transfer of plasmids was studied under laboratory conditions and in an open environment during the utilization of polycyclic aromatic hydrocarbons. The rhizospheric bacteria of the genus Pseudomonas belonging to the unique PGPR group were studied. The strains produce a wide range of biologically active metabolites, including siderophores, phenazines, 2,4-diacetylphloroglucin, pyoluteorin, pyrrolnitrin, oomycin A, hydrocyanic acid, synthesize phytohormones, including indolyl-3-acetic acid (IAA), and stimulate root formation in plants. Based on one of these strains of P. chlororaphis (formerly P. aureofaciens) BS1393, the biopreparation Pseudobacterin-2 was developed, which is highly effective against a number of plant diseases.

Mass spectrometry was applied to identify metabolites and estimate the efficiency of phenanthrene biodegradation and transformation by rhizosphere bacteria Pseudomonas aureofaciens BS1393. Strains P. aureofaciens BS1393(pOV17) and P. aureofaciens BS1393(NPL-41) bearing various naphthalene biodegradation plasmids were used in the work. The strain BS1393(pOV17) contains the pOV17 wild type naphthalene biodegradation plasmid that determines the oxidation of naphthalene to Krebs cycle metabolites. The strain BS1393(NPL-41) contains the mutant plasmid NPL-41 governing the initial stages of naphthalene oxidation into salicylic acid. The limiting stages of phenanthrene biodegradation in bacteria with various plasmids have been identified according to the accumulation of intermediates. When bacteria were grown on phenanthrene, the main metabolites were as follows: (a) 2-hydroxy-2H-benzo[h]chromene-2-carboxylic acid/trans-4-(1-hydroxynaph-2-yl)-2-oxobut-3-enoic acid, (b) 1-hydroxy-2-naphthoic acid, and (c) salicylic acid. In the strain BS1393(pOV17), metabolite (а) was observed during 1–14 days of cultivation. Unlike it, in the strain BS1393(NPL-41), an insignificant amount of this metabolite was found after only 14 days. The availability of metabolite (b) in the growth of both strains was an evidence of the limited rate of its further decarboxylation. Metabolite (c) as a final product was found in the growth of the strain BS1393(NPL-41). Contrastingly, in the strain BS1393(pOV17), this metabolite was not found, which indicates the complete oxidation of phenanthrene.

The dynamics of wolf attacks on domestic ungulates at the levels of population group (the region of Central Forest Nature Reserve) and population (Tver oblast) have been analyzed over the period of 1971 to 2015. It has been found that the frequency of attacks increases during the phase of predator population growth under the effect of qualitative changes within the population. An increase in the frequency of attacks and the number of sheep and calves killed per attack also takes place in the summer–autumn period, as a consequence of training wolf cubs in methods of killing prey. Therefore, domestic animals are not only a source of food for the wolf but also an object for training the young.

Major metabolites formed by Pseudomonas bacteria bearing various naphthalene degradation plasmids were identified using high performance liquid chromatography and mass spectrometry in combination with electrospray ionization. During the growth of P. aureofaciens BS1393(NPL-41) bacteria bearing the plasmid NPL-41, partial oxidation of naphthalene was shown to occur to form the major exometabolite (salicylate) and minor exometabolite (phenol). The growth of P. aureofaciens BS1393(pOV17) bacteria bearing the plasmid pOV17 was accompanied by complete oxidation of naphthalene and formation of trace amounts of salicylate and epoxybenzene. Based on the data of molecular mass spectrometry, we determined the dynamics of CO2 production and oxygen uptake in the process of bacterial growth. The specific quantity of CO2 per unit of cell amount for P. aureofaciens BS1393(NPL-41) bacteria was shown to exceed fivefold that for P. aureofaciens BS1393(pOV17). The respiratory quotient (RQ) representing the ratio of the molar concentrations of metabolic CO2 to consumed oxygen depends on the type of plasmids that determine naphthalene degradation. The value of RQ during the growth of P. aureofaciens BS1393(pOV17) bacteria was almost 1.5 times greater than in the case of P. aureofaciens BS1393(NPL-41). The ratio of O2 uptake to CO2 formed by the first culture was 4.5 mol O2/mol CO2; by the second culture, 6.5 mol O2/mol CO2.

This paper studies the manifestation of philopatry and dispersal in wolves at the level of family, population group (the Central Forest Reserve), and population (Tver oblast). The importance of a den area for a wolf in the formation of the territorial structure is shown. The “philopatric” behavior of 35 wolves from seven families in a completing circle of flags is analyzed: 32 individuals (16 adult and 16 young wolves from different families) crossed flags toward a den, and only three chose a different direction. This behavior was typical for the entire population group of wolves both for young individuals born and raised in this place and for adult wolves (both resident wolves and their children who have created here new pairs and then families). Attention is focused on the role of “dispersant” wolves in 1972–1982: a rapid increase in the population was caused by internal reserves due to the dispersal of juveniles for short distances. Analysis of the available field and literature data shows that dispersers “transfer” ecological, behavioral, spatial functional, and structural features of a family; i.e., they are bearers of its traditions.

By the example of glucose uptake by the soil bacteria Pseudomonas aureofaciens BS1393(pBS216) and Rhodococcus sp. 3–30 immobilized on a solid-phase surface (quartz sand), their growth parameters were determined: growth rate (doubling time), total CO2 production, CO2 production per cell, lag period with respect to substrate uptake, respiratory quotient. The growth of P. aureofaciens and Rhodococcus sp. on glucose revealed (1) differences of the lag period with respect to substrate (lag time of ∼4 h for P. aureofaciens and ∼26 h for Rhodococcus sp.); (2) differences between the maximal rates of CO2 production (∼50 μg C-CO2 g−1 sand h−1 for P. aureofaciens and ∼8.5 μg C-CO2 g−1 sand h−1 for Rhodococcus sp.); (3) differences in CO2 production per cell (∼1.94 × 10−9 μM CO2/CFU for P. aureofaciens and more than ∼3.4 × 10−9 μM CO2/CFU for Rhodococcus sp.). The kinetics of the metabolic CO2 isotopic composition was shown to be determined by the difference in the carbon isotopic characteristics of products in the cell. Upon introduction of glucose into the medium (the preparatory stage of the metabolism), the uptake of intracellular 13C-depleted products (lipids) is noted; at the stage of the maximal cell growth rate, introduced glucose is mainly metabolized; and at the final stage, upon exhaustion of substrate, the “stored” products—the lipid fraction—get involved in the metabolism. At the maximal rate of glucose uptake, the CO2 carbon isotopic fractionation coefficient relative to organic products of microbial biosynthesis was determined to be α = 1.009 ± 0.002.

Using molecular and isotopic mass spectrometry, we investigated the toxic effect of naphthalene as a representative of polycyclic aromatic hydrocarbons (PAHs) on plants growing under sterile conditions and plants inoculated with microorganisms capable and incapable of naphthalene degradation. Tobacco plants of the Samsun variety were grown in a closed gas-nutrient system on a mineral medium with sucrose as a carbon source. Naphthalene used as a toxicant at a concentration of 5.2 × 10 −4 % contained 13 C isotope whose amount was characterized by the value δ 13 C = +281.4 ± 0.6‰ relative to the PDB standard and differed from that of sucrose, the main source of carbon (δ 13 C = −12.0 ± 0.1‰). Degradation of naphthalene was determined by the inclusion of its carbon in metabolic CO 2 and plant tissues (the root, stem, leaves). The effect of naphthalene on plants was indicated by the rates of O 2 production and CO 2 uptake during the light period as compared with the dark period of exposure. A decrease of the toxic effect of naphthalene on plants was observed only at the inoculation of plants with Pseudomonas aureofaciens BS1393 rhizosphere bacteria bearing plasmid pBS216, which controls the naphthalene biodegradation ability. The occurrence of other heterotrophic microorganisms incapable of naphthalene degradation had no similar protective effect.

На примере потребления глюкозы почвенными бактериями Pseudomonas aureofaciens BS1393(pBS216) и Rhodococcus sp. 3-30, иммобилизованными на твердофазной поверхности (кварцевый песок), определены их ростовые показатели: скорость роста бактерий (время удвоения клеток), суммарная продукция СО2 и продукция из расчета на бактериальную клетку, лаг-период относительно потребления субстрата, дыхательный коэффициент. При росте бактерий P. aureofaciens и Rhodococcus sp. на глюкозе выявлены различия штаммов по параметрам: (1) лаг-периодов относительно внесенного субстрата (лаг-период около 4 ч для P. aureofaciens и около 26 ч для Rhodococcus sp.); (2) максимальной скорости продукции СО2 (около 50 мкг СО2 г-1 песка ч-1 для P. aureofaciens и около 8.5 мкг СО2 г-1 песка ч-1 для Rhodococcus sp.); (3) продукции СО2 из расчета на одну клетку (около 1.94 ? 10-9 мкмоль СО2 для P. aureofaciens и около 3.4 ? 10-9 мкмоль СО2 для Rhodococcus sp.). Показано, что кинетика изотопного состава метаболической СО2 определяется различием в изотопных характеристиках углерода продуктов в клетке: после внесения глюкозы в среду (подготовительная стадия метаболизма) отмечено использование внутриклеточных обедненных 13 продуктов (липидов), на стадии максимальной скорости роста клеток метаболизируется, главным образом, внесенная глюкоза, и на завершающей стадии после исчерпания субстрата в обмен включаются “запасенные” продукты – липидная фракция. При максимальной скорости потребления глюкозы определен коэффициент фракционирования изотопов углерода СО2 относительно органических продуктов микробного биосинтеза, который составляет величину = 1.009 ± 0.002.

The in vivo and in vitro interactions between tobacco (Nicotiana tabacum L.), tomato (Lycopersicon esculentum Mill.), cabbage (Brassica oleracea var. capitata L.), rape (B. napus L.), and the common ice plant (Mesembryanthemum crystallinum L.) and bacteria Pseudomonas aureofaciens, P. putida, and Methylovorus mays were studied. Stable associations of these microorganisms with plants are demonstrated. Colonized plants were characterized by accelerated growth, more efficient rooting, better adaptation to in vivo conditions, and enhanced resistance to bacterial and fungal phytopathogens (Erwinia carotovora, Sclerotinia sclerotiorum and Phytophthora infestans). Plants colonized by bacteria resistant to kanamycin and naphthalene can grow steadily on the medium containing these compounds. The results obtained indicate a promising usage of beneficial associative microorganisms for the development of technologies for plant protection against biotic and abiotic stressors.

Исследован углеводородокисляющий потенциал почвенной микробиоты и интродуцированных в почву углеводородокисляющих микроорганизмов на основе количественных и изотопных характеристик углерода продуктов, образующихся при микробной деградации нефти. Из сравнения скоростей продукции СО2 в нативной почве и почве, загрязненной сырой нефтью, обнаружено, что интенсивность микробной минерализации почвенного органического вещества (ПОВ) в присутствии нефти выше по сравнению с незагрязненной почвой, т.е., обнаруживается затравочное влияние (прайминг-эффект) углеводородов нефти. Показано, что количество углерода вновь синтезированных органических продуктов за счет потребленной нефти (биомасса клеток и экзометаболиты) значительно превосходит количество ПОВ, израсходованное на продукцию СО2. Обнаружено, что в результате микробиологических процессов в почве, загрязненной нефтью, наблюдается мощный поток углекислоты, поступающей в атмосферу.

The hydrocarbon-oxidizing potential of soil microbiota and hydrocarbon-oxidizing microorganisms introduced into soil was studied based on the quantitative and isotopic characteristics of carbon in products formed in microbial degradation of oil hydrocarbons. Comparison of CO2 production rates in native soil and that polluted with crude oil showed the intensity of microbial mineralization of soil organic matter (SOM) in the presence of oil hydrocarbons to be higher as compared with non-polluted soil, that is, revealed a priming effect of oil. The amount of carbon of newly synthesized organic products (cell biomass and exometabolites) due to consumed petroleum was shown to significantly exceed that of SOM consumed for production of CO2. The result of microbial processes in oil-polluted soil was found to be a potent release of carbon dioxide to the atmosphere.

The complete sequence of the 46,267 bp genome of the lytic bacteriophage tf specific to Pseudomonas putida PpG1 has been determined. The phage genome has two sets of convergently transcribed genes and 186 bp long direct terminal repeats. The overall genomic architecture of the tf phage is similar to that of the previously described Pseudomonas aeruginosa phages PaP3, LUZ24 and phiMR299-2, and 39 out of the 72 products of predicted tf open reading frames have orthologs in these phages. Accordingly, tf was classified as belonging to the LUZ24-like bacteriophage group. However, taking into account very low homology levels between tf DNA and that of the other phages, tf should be considered as an evolutionary divergent member of the group. Two distinguishing features not reported for other members of the group were found in the tf genome. Firstly, a unique end structure - a blunt right end and a 4-nucleotide 3'-protruding left end - was observed. Secondly, 14 single-chain interruptions (nicks) were found in the top strand of the tf DNA. All nicks were mapped within a consensus sequence 5'-TACT/RTGMC-3'. Two nicks were analyzed in detail and were shown to be present in more than 90% of the phage population. Although localized nicks were previously found only in the DNA of T5-like and phiKMV-like phages, it seems increasingly likely that this enigmatic structural feature is common to various other bacteriophages.