Investigation of Lysozyme–Antilysozyme Interactions in a Model Tetrahymena–EscherichiaCommunity

Lysozyme and antilysozyme activities present in a wide range of microorganisms determine the so-called lysozyme–antilysozyme system of hydrobionts, which greatly contribute to the formation of aquatic biocenoses. However, the mechanism of the functioning of this system in natural freshwater communities remains obscure. The experimental investigation of lysozyme–antilysozyme interactions in a model Tetrahymena–Escherichiacommunity showed that the antilysozyme activity of Escherichia colileads to incomplete phagocytosis, thus enhancing bacterial survival in a mixed culture with infusoria. The selection and reproduction of bacterial cells resistant to grazing by infusoria determine the character of host–parasite interactions and allow bacteria to survive. It was demonstrated that the antilysozyme activity of microorganisms, which is responsible for bacterial persistency in natural biocenoses, is involved in the maintenance of protozoa–bacteria communities in bodies of water.     

Enzyme-substrate mode of action for bacteria survival in water biotic community

The role of enzyme--substrate interactions for the survival of bacteria in natural biocenoses has been analyzed with the systems "lysozyme-antilysozyme", "histon-antihiston" used as models. The conception of a possible universal mechanism for supporting the circulation of pathogenic and opportunistic microorganisms among eukaryotes, irrespective of their evolutionary status and environment, has been formulated. This mechanism is ensured by the natural resistance of the eukaryotic cells and by persistence factors on the part of the pathogen; as a result, the dynamic system of interactions is formed which facilitates the survival of microorganisms due to their persistence potential. New knowledge thus obtained opens prospects in the study of sanitary and ecological aspects of water biocenoses.     

Significance of hydrobiont persistent properties for symbiotic interactions

Significance of symbiotic relations formed by associative symbiosis type for autochthonous and allochthonous microflora of natural water bodies is shown. Generality of symbiotic interaction mechanisms of symbionts in limnetic and halophilous communities provided by secreted factors of natural resistance from the side of the host, and by factors of persistence from the side of symbionts is proven based on a set of examples. Features of operation of lysozyme-antilysozyme, histon-antihiston, hydrogen peroxide-catalase functional systems in symbiotic interactions of autotrophic and heterotrophic components of hydrobiocenosis with dominant and associative microflora are presented. Associative microflora of allochthonous origin was shown to actively use the ecologically formed system of interaction between hydrobionts that facilitates survival of these microorganisms and preservation of their persistent potential, and as a result leads to biocenosis disorders. The knowledge obtained open new possibilities and perspectives of research of sanitary and ecological aspects of vital activity of aquatic biocenoses.     

Interaction of a causative agent with associative bacteria during salmonellosis

AIM: To determine a composition of gut microflora during salmonellosis and to study the modification of persistent characteristics (antilysozyme activity, ALA) of symbiotic microorganisms in associations. MATERIALS AND METHODS: Bacteriologic study of feces was performed in 90 patients aged 18-39 years, which were divided to three groups: patients with salmonellosis in acute phase, reconvalescent patients, and conditionally healthy persons. Condition of gut microflorawas determined; microorganisms associated with Salmonella infection were isolated, and their influence on ALA of Salmonella was studied. RESULTS: Gut microbiocenosis was more diverse in patients compared with healthy persons. Significant reduction of bifidobacteria quantity (to 10(7) CFU/g of feces and less), especially in reconvalescent period, was noted. Association between bifidoflora deficiency and excessive increase of quantity of yeast fungi was revealed. It was determined that exometabolites of indigenous anaerobic microflora (bifidobacteria) promoted decrease of ALA of Salmonella, whereas opportunistic facultatively anaerobic microorganisms (enterobacteria, staphylococci) rendered mainly stimulating effect on the ALA of Salmonella. CONCLUSION: Obtained data reveal characteristics of bacterial interactions in associative symbiosis and provide insights about mechanisms of formation of pathobiocenosis and state of bacterial carriage.     

Surface Water Linkages Regulate Trophic Interactions in a Groundwater Food Web

Groundwaters are increasingly viewed as resource-limited ecosystems in which fluxes of dissolved organic carbon (DOC) from surface water are efficiently mineralized by a consortium of microorganisms which are grazed by invertebrates. We tested for the effect of groundwater recharge on resource supply and trophic interactions by measuring physico-chemistry, microbial activity and biomass, structure of bacterial communities and invertebrate density at three sites intensively recharged with surface water. Comparison of measurements made in recharge and control well clusters at each site showed that groundwater recharge significantly increased fluxes of DOC and phosphate, elevated groundwater temperature, and diminished dissolved oxygen (DO). Microbial biomass and activity were significantly higher in recharge well clusters but stimulation of autochthonous microorganisms was not associated with a major shift in bacterial community structure. Invertebrate assemblages were not significantly more abundant in recharge well clusters and did not show any relationship with microbial biomass and activity. Microbial communities were bottom-up regulated by DOC and nutrient fluxes but trophic interactions between microorganisms and invertebrates were apparently limited by environmental stresses, particularly DO depletion and groundwater warming. Hydrological connectivity is a key factor regulating the function of DOC-based groundwater food webs as it influences both resource availability for microorganisms and environmental stresses which affect energy transfer to invertebrates and top-down control on microorganisms.     

The antilysozyme activity of meningococci

The antilysozyme activity of 184 meningococcal strains was studied. Such activity was found in all strains within the range 1-25 micrograms/ml, which was due to the heterogeneity of bacterial population. Strains varying in the total level of their lysozyme activity differed in the population structure by this sign. Preparations inhibiting antilysozyme activity could be differentiated according to the character of their action by the method of the population analysis. In the process of phagocytosis the selection of clones with a high level of antilysozyme activity was found to occur. The antilysozyme factor, heat resistant protein with a molecular weight of 10,000-30,000 D, was sensitive to trypsin.     

Circadian rhythms of antibacterial resistance, coagulase and antilysozyme activity of Staphylococcus aureus

AIM: To determine daily dynamics of antibacterial resistance as well as antilysozyme and coagulase activity of S. aureus strains. MATERIALS AND METHODS: On an example of clinical strains of S.aureus isolated from patients with surgical infections daily dynamics of biological characteristics of staphylococci was studied. After 12 hours of incubation strains were tested for coagulase activity by standard method (test tube method), antilysozyme activity by photometric method, and antibacterial resistance by method of serial dilutions in agar. Tests were repeated each 3-hours during a day. RESULTS: Variation of levels of studied biological characteristics of staphylococci during a day was revealed. Structures of coagulase and antilysozyme circadian rhythms had some differences in different S. aureus strains. Alongside with it, similarity in temporal expression of such biological characteristics of staphylococci as antibacterial resistance and antilysozyme activity was noted. CONCLUSION: Obtained data open prospect to use biorhythmological approach in study of biological characteristics of microorganisms during evaluation of their mechanisms of adaptation to changing environmental conditions. Chronobiological approach allows to reveal periods of maximal expression of S. aureus characteristics that could be used for increasing of effectiveness of antibacterial treatment by the choice of optimal time for administration of antibiotic.     

Method of determining the antilysozyme activity of microorganisms

The method for the determination of the antilysozyme activity of microorganisms is described. The method consists in the cultivation of the strains under study in a lysozyme-containing medium, and the effect of lysozyme inactivation is determined from the growth of Micrococcus luteu S indicator strain adjacent to active strains. The quantitative evaluation of this property is presented. The study of 1 296 strains belonging to 9 genera has disclosed that antilysozyme activity occurs most frequently among Gram-negative bacteria.     

Experimental evaluation of interaction between Yersinia pestis and soil infusoria and possibility of prolonged preservation of bacteria in the protozoan oocysts

The character and outcome of interactions between Y. pestis (vaccine strain and soil infusoria Tetrahymena pyriformis (axenic culture) were under experimental study. The parallel use of the bacteriological method and PCR test systems made it possible to follow the dynamics of Y. pestis cells (strain EV) with different plasmid profiles in their interaction with infusoria, as well as their passage into the protozoa cysts. The study revealed the complete utilization of Y. pestis cells lacking virulence factors by infusoria. The presence of plasmids of virulence influenced only the duration of complete bacterial phagocytosis. A drop in the temperature of cultivation to 2 degrees C induced the mass and rapid encystment of infusoria. In the PCR analysis specific DNA fragments of Y. pestis cells, preserved in the latent (uncultivable) state, were detected in the cysts of protozoa within the period of up to 14 months, while the results of bacteriological studies were negative. The data thus obtained are discussed with regard to the possible mechanisms of the existence and prolonged reservation of Y. pestis in the soils of natural foci with participation of protozoa.     

Features of soil nematode communities in various types of natural biocenoses: Effectiveness of assessment parameters

The relations of different characteristics of soil nematode communities with the type of natural (forest and meadow) biocenoses and the main parameters of assessment of the state of soil ecosystems were determined in the course of monitoring. It was found that the number of nematodes and number of taxa in the fauna are highly variable. It was noted that meadow biocenoses stand apart by the diversity of nematodes and the proportion of plant parasites in the community structure. A significant functional contribution in the distinguishing of the main types of biocenoses was determined only for the index reflecting the organic matter decomposition in the soil using discriminant analysis. It was shown that the complex analysis of the parameters of nematode communities most informatively characterizes the state of the soil ecosystem.     

Increasing antagonistic interactions cause bacterial communities to collapse at high diversity

Biodiversity is a major determinant of ecosystem functioning. Species-rich communities often use resources more efficiently thereby improving community performance. However, high competition within diverse communities may also reduce community functioning. We manipulated the genotypic diversity of Pseudomonas fluorescens communities, a plant mutualistic species inhibiting pathogens. We measured antagonistic interactions in vitro, and related these interactions to bacterial community productivity (root colonisation) and ecosystem service (host plant protection). Antagonistic interactions increased disproportionally with species richness. Mutual poisoning between competitors lead to a 'negative complementarity effect', causing a decrease in bacterial density by up to 98% in diverse communities and a complete loss of plant protection. The results emphasize that antagonistic interactions may determine community functioning and cause negative biodiversity-ecosystem functioning relationships. Interference competition may thus be an additional key for predicting the dynamics and performance of natural assemblages and needs to be implemented in future biodiversity models.     

Considerable Bacterial Community Structure Coupling with Extracellular Electron Transfer at Karst Area Stone in Yunnan,China

Nearly all karst stones have partial dark and light color sections under the same condition. In this study, 24662 operational taxonomic units (OTUs) were examined from karst stones surface samples. Dark samples showed higher abundance of 18,115 OTUs (73.5%) and containing clustered bacterial communities, as indicated by the results of principal component and cluster analyses. Additionally, heatmap analysis showed microorganisms distinction between different color samples and 19 genera of electroactive microorganisms gathered in the dark samples. Furthermore, iron manganese oxides were detected as the main mineral composition difference. Notably, dark samples exhibited remarkable photoelectrochemical activity in response to visible light. Under sunlight, both microorganisms and minerals displayed close relationship with extracellular electron transfer process, which enhanced the understanding for microorganism-–mineral interactions in natural karst environments.     

Persistent properties of protozoa-associated bacteria

The species structure and persistent properties (antilysozyme and antihistone activity) of bacteria forming associations with protozoa is revealed. Among them, 68.9% of the isolates were enterobacteria, the remaining organisms belonged to the families Aeromonas, Alcaligenes, Pseudomonas, Vibrio, etc. Within the family Enterobacteriaceae bacteria of the Escherichia group prevailed. 50.4% of the isolates were found to have antilysozyme activity and 97%--antihistone activity. The level of persistent properties in the representatives of allochthonous microflora was higher than that in the representatives of autochthonous microflora. In addition to antilysozyme activity antihistone activity was noted in protozoa-associated bacteria, which could be of importance for the formation of symbiotic links in natural associations. These data may be used in sanitary and hygienic practice for microecological monitoring of the environment.     

Effect of vaginal Lactobacillus metabolites on biological properties of Staphylococcus epidermidis and Escherichia coli

The influence of supernatants and cell extracts of vaginal lactic acid bacilli on the growth, catalase, antilysozyme and anticomplementary activity of S. epidermidis and E. coli was studied. Changes observed in the persistence characteristics of these microorganisms were found to develop in different directions: in S. epidermidis a decrease in antilysozyme activity was mainly observed, while in E. coli the suppression of anticomplementary activity was registered. The modifying influence of lactic acid flora, revealed in this investigation, is regarded as one of the mechanisms facilitating the formation of microbiocenosis and ensuring colonization resistance.     

General and molecular ecology of Legionella

The review is devoted to the general and molecular ecology of bacteria of the genus Legionella in natural and anthropogenic environments. Invasion of amoebae and infusoria by legionellae and their replication in these protozoa can be considered to be a pre-adaptation for invasion of the human immune system. Symbiosis of bacteria and protozoa as a promising model of cellular microbiology and the conception of bacterial ecological niches are discussed in relation to the low fidelity of most bacterial species to their habitats (biotopes). The necessity of elaboration of a similar conception for microbial consortia and associations is emphasized.     

General and molecular ecology ofLegionella

The review is devoted to the general and molecular ecology of bacteria of the genusLegionella in natural and anthropogenic environments. Invasion of amoebae and infusoria by legionellae and their replication in these protozoa can be considered to be a preadaptation for invasion of the human immune system. Symbiosis of bacteria and protozoa as a promising model of cellular microbiology and the conception of bacterial ecological niches are discussed in relation to the low fidelity of most bacterial species to their habitats (biotopes). The necessity of elaboration of a similar conception for microbial consortia and associations is emphasized.     

Persistence factors of Francisella tularensis

The study of the persistence potential of 64 F. tularensis strains isolated from different sources was carried out. The wide spread of the antilysozyme, antilactoferrin and anticomplementory activities of F. tularensis were detected. F. tularensis, isolated from ticks and water, were characterized by the highest level of the expression of antilysozyme activity, while anticomplementory and antilactoferrin activities of the infective agents were characteristic of those microorganisms which were isolated from rodents and their excrements.     

DNA-based monitoring of total bacterial community structure in environmental samples

Determining the structure of bacterial communities and their response to stimuli is key to understanding community function and the interactions that occur between microorganisms and the environment. However, bacterial communities often comprise complex assemblages of large numbers of different bacterial populations. An approach is presented which allows bacterial community structure to be determined by fractionation of the complex mixture of total bacterial community DNA using the DNA-binding dye bisbenzimidazole which imposes G + C-dependent changes in the buoyant density of DNA. Bacterial community structure presented as percentage of total DNA vs. percentage G + C content of DNA is an indication of the relative abundance of phylogenetic groups of bacteria. Changes in the composition of a soil bacterial community in response to perturbations in the form of carbon amendment and altered water status were monitored.