Biocenosis of the natural foci of sapronotic infections (the results of 15-year observations)

The summarized results of original studies, carried out in 1988-2003 by scientists of the Gamaleya Research Institute of Epidemiology and Microbiology, are discussed. The part dealing with the regularity and mechanisms of the circulation of pathogenic bacteria in the biocenosis of soil and water reservoirs includes the following subjects: population and intracellular interactions with protozoa; the evaluation of different representatives of hydrobios as hosts and the transmission along trophic biocenotic chains; the effect of algae and their metabolic products on bacterial populations; the possibility of the colonization of higher plants from the soil and the subsequent infection of rodents; the variability and clonal structure of bacterial population in its interaction with protozoa and plants; the chain of Yersinia circulation in agrocenosis. The part dealing with the mechanisms of the prolonged reservation of causative agents includes such subjects as the reversible transition of bacteria into the latent (uncultivable) state in soils and water reservoirs, as well as the biological inductors of this process; the prolonged preservation of latent bacteria in the cysts of protozoa and blue-green algae; the indication of causative agents in the natural foci of plague and pseudotuberculosis. On the basis of the original investigations and the data of literature new theoretical review in the field of the natural foci and the epidemiology of sapronotic infections have been formulated.     

An extracellular Leptospira interrogans leucine‐rich repeat protein binds human E‐ and VE‐cadherins

Pathogenic Leptospira bacteria are the causative agents of leptospirosis, a zoonotic disease affecting animals and humans worldwide. These pathogenic species have the ability to rapidly cross host tissue barriers by a yet unknown mechanism. A comparative analysis of pathogens and saprophytes revealed a higher abundance of genes encoding proteins with leucine‐rich repeat (LRR) domains in the genomes of pathogens. In other bacterial pathogens, proteins with LRR domains have been shown to be involved in mediating host cell attachment and invasion. One protein from the pathogenic species Leptospira interrogans, LIC10831, has been previously analysed via X‐ray crystallography, with findings suggesting it may be an important bacterial adhesin. Herein we show that LIC10831 elicits an antibody response in infected animals, is actively secreted by the bacterium, and binds human E‐ and VE‐cadherins. These results provide biochemical and cellular evidences of LRR protein‐mediated host–pathogen interactions and identify a new multireceptor binding protein from this infectious Leptospira species.     

Planktonic food chains of a highly humic lake

The development and metabolism of the plankton of a highly humic lake were followed over the vernal primary production maximum. The study was made in a mesocosm in which large filter feeders, typical of this lake in summer, were absent. During the rising phase of phytoplankton, the community was predominantly autotrophic. The most important constituents in the algal biomass were a dinoflagellate, Gymnodinium sp. (40–50%), and a prasinophycean, Scourfieldia cordiformis (7%). The biomasses of Chlamydomonas spp. and Chrysococcus spp. reached their maxima a few days later and Cryptomonas sp. became most abundant at the end of the experiment. After the phytoplankton maximum, about one week from the beginning ofthe experiment, grazing of algae by phagotrophic protozoans and phosphate depletion led to a rapid decrease of algal biomass and the community became predominantly heterotrophic. In spite of a large variation in algal biomass and primary production, the biomass of bacteria remained of the same order of magnitude as in algae both before and after the algal maximum. Bacteria were mostly responsible for the plankton respiration, which also showed no dependence on primary production. Since exudation by phytoplankton was also low, the nutrition of bacterioplankton was probably mainly based on allochthonous dissolved organic matter rather than or primary production. Thus the production of bacteria was an additional food source for higher trophic levels along with phytoplankton. Because filter feeding zooplankton was absent in the experiment, protozoans were the only grazers utilizing algae and bacteria. Essentially all growth of bacteria was used by bacterivores.     

Natural-focus infections: the key questions and new vantage points

A short history of the concept of natural focal infections is presented: the idea put forward by D. K. Zabolotny?, E. I. Pavlovski?'s teaching, 3 stages of its development. A number of fundamental questions and the modern content of the concept are considered. The natural foci of infections are a combination of surface, soil and/or water ecosystems, including the population of the causative agent of infection. In contrast to surface ecosystems, in soil and water ecosystems the hosts of the causative agents of sapronotic infections ae soil invertebrates and hydrobios, in which these agents may circulate in biocenotic trophic chains. The circulation of the causative agents in natural foci is a discrete process; the mechanisms and forms of the existence of pathogenic bacteria during seasonal and prolonged periods between epidemics is considered. Special attention is given to latent (nonculturable) forms of bacteria. The complex character of the status of the causative agents of natural focal infections is discussed.     

Role of lactic acid bacteria in the spread of antibiotic resistant bacteria among healthy persons

The wide use of antibiotics in livestock raising has contributed to the selection and accumulation of representatives of commensal microflora, as well as pathogenic bacteria, colonizing livestock and poultry. For this reason the problem of the possible transfer of antibiotic-resistance genes along the chain from bacteria, autochthonous for agricultural animals, to bacteria used for the production of foodstuffs, which are incorporated into normal microflora and may thus participate in the exchange of these genes with bacteria, enteropathogenic for humans, is a highly important task of medical microbiology. The article deals with the review of experimental data, indicative the possibility of the appearance of antibiotic-resistant pathogenic bacteria due to the transfer of antibiotic-resistance genes via alimentary chains.     

Animal infection models using non-mammals

The use of non-human animal models for infection experiments is important for investigating the infectious processes of human pathogenic bacteria at the molecular level. Mammals, such as mice and rabbits, are also utilized as animal infection models, but large numbers of animals are needed for these experiments, which is costly, and fraught with ethical issues. Various non-mammalian animal infection models have been used to investigate the molecular mechanisms of various human pathogenic bacteria, including Staphylococcus aureus, Streptococcus pyogenes, and Pseudomonas aeruginosa. This review discusses the desirable characteristics of non-mammalian infection models and describes recent non-mammalian infection models that utilize Caenorhabditis elegans, silkworm, fruit fly, zebrafish, two-spotted cricket, hornworm, and waxworm.     

Omnivory and the stability of simple food webs

Traditional ecological theory predicts that the stability of simple food webs will decline with an increasing number of trophic levels and increasing amounts of omnivory. These ideas have been tested using protozoans in laboratory microcosms. However, the results are equivocal, and contrary to expectation, omnivory is common in natural food webs. Two recent developments lead us to re-evaluate these predictions using food webs assembled from protists and bacteria. First, recent modelling work suggests that omnivory is actually stabilizing, providing that interactions are not too strong. Second, it is difficult to evaluate the degree of omnivory of some protozoan species without explicit experimental tests. This study used seven species of ciliated protozoa and a mixed bacterial flora to assemble four food webs with two trophic levels, and four webs with three trophic levels. Protist species were assigned a rank for their degree of omnivory using information in the literature and the results of experiments that tested whether the starvation rate of predators was influenced by the amount of bacteria on which they may have fed and whether cannibalism (a form of omnivory) occurred. Consistent with recent modelling work, both bacterivorous and predatory species with higher degrees of omnivory showed more stable dynamics, measured using time until extinction and the temporal variability of population density. Systems with two protist species were less persistent than systems with one protist species, supporting the prediction that longer food chains will be less stable dynamically. Received: 28 December 1997 / Accepted: 22 June 1998     

The role of quorum sensing mediated developmental traits in the resistance of Serratia marcescens biofilms against protozoan grazing

Resistance against protozoan grazers is a crucial factor that is important for the survival of many bacteria in their natural environment. However, the basis of resistance to protozoans and how resistance factors are regulated is poorly understood. In part, resistance may be due to biofilm formation, which is known to protect bacteria from environmental stress conditions. The ubiquitous organism Serratia marcescens uses quorum sensing (QS) control to regulate virulence factor expression and biofilm formation. We hypothesized that the QS system of S. marcescens also regulates mechanisms that protect biofilms against protozoan grazing. To investigate this hypothesis, we compared the interactions of wild-type and QS mutant strains of S. marcescens biofilms with two protozoans having different feeding types under batch and flow conditions. Under batch conditions, S. marcescens forms microcolony biofilms, and filamentous biofilms are formed under flow conditions. The microcolony-type biofilms were protected from grazing by the suspension feeder, flagellate Bodo saltans, but were not protected from the surface feeder, Acanthamoeba polyphaga. In contrast, the filamentous biofilm provided protection against A. polyphaga. The main findings presented in this study suggest that (i) the QS system is not involved in grazing resistance of S. marcescens microcolony-type biofilms; (ii) QS in S. marcescens regulates antiprotozoan factor(s) that do not interfere with the grazing efficiency of the protozoans; and (iii) QS-controlled, biofilm-specific differentiation of filaments and cell chains in biofilms of S. marcescens provides an efficient mechanism against protozoan grazing.     

Response of Heterotrophic Planktonic Bacteria to the Zebra Mussel Invasion of the Tidal Freshwater Hudson River

Abstract Invasions of aquatic ecosystems by exotic bivalves are known to cause dramatic changes in phytoplankton and some other groups, but their effect on the microbial component is unknown. The invasion of the tidal freshwater Hudson River by the exotic zebra mussel (Dreissena polymorpha) has caused large changes in several components of the Hudson's food web. Planktonic bacteria in the tidal freshwater Hudson are a major part of the food web, and mediate important processes in the carbon budget. We used a long-term data set, spanning four years prior to the zebra mussel (ZM) invasion and four years post-invasion, to describe ZM effects on planktonic bacteria. Small and meso-scale experiments were conducted to specifically examine direct consumption of bacteria by ZM, as well as effects on protozoans. Bacterial abundances in the Hudson have increased roughly 2× since the ZM arrived, making it clear that direct consumption by Dreissena is a minor process. Experiments show that ZM do not remove bacteria from Hudson River water, but are very effective at clearing flagellated protozoans, the major predator of bacteria. The observed changes in bacterial abundance have not been accompanied by equally large changes in bacterial productivity, suggesting growth is primarily limited by carbon supply. Bacterial production has not declined despite a dramatic decline of phytoplankton, confirming previous suggestions that bacteria and phytoplankton are not strongly linked in the Hudson. As a result of the increase in bacterial abundance and removal of phytoplankton, the absolute and relative contributions of bacterial carbon to living particulate organic carbon (POC) standing stocks have increased dramatically. The maintenance of the bacterial component of the Hudson River's food web may be one mechanism whereby consumers are ``insulated' from effects of zebra mussel consumption of phytoplankton carbon. Received: 24 October 1997; Accepted: 9 February 1998     

<Emphasis Type="Italic">Coxiella</Emphasis> Symbionts in the Cayenne Tick <Emphasis Type="Italic">Amblyomma cajennense</Emphasis>

Members of the Coxiella genus are intracellular bacteria that can infect a variety of animals including humans. A symbiotic Coxiella was recently described in Amblyomma americanum ticks in the Northern Hemisphere with no further investigations of other Amblyomma species in other geographic regions. These ixodid ticks represent a group of important vectors for human infectious agents. In the present work, we have demonstrated that symbiotic Coxiella (SCox) are widespread, occurring in South America and infecting 100% of all life stages and eggs of the Cayenne ticks Amblyomma cajennense from Brazil and the USA. Using light microscopy, in situ hybridization, and PCR, we demonstrated SCox in salivary glands, ovaries, and the intestines of A. cajennense. These symbionts are vertically and transtadially transmitted in laboratory reared A. cajennense, and quantitative PCR analyses indicate that SCox are more abundant in adult female ticks, reaching values corresponding to an 11×, 38×, and 200× increase in SCox 16S rRNA gene copy number in unfed females, compared to unfed nymphs, larvae, and eggs, respectively. Phylogenetic analyses showed distinct SCox subpopulations in the USA and Brazil and demonstrated that SCox bacteria do not group with pathogenic Coxiella burnetii.     

Comparison of pelagic food webs in lakes along a trophic gradient and with seasonal aspects: influence of resource and predation

Composition and seasonal dynamics of phytoplankton, bacteria,and zooplankton (including heterotrophic flagellates, ciliates,rotifers and crustaceans) were studied in 55 lakes in NorthernGermany with different trophic status, ranging from mesotrophicto hypertrophic. Mean abundance and biomass of all groups increasedsignificantly with trophic level of the lake, but bacteria andmetazooplankton showed only a weak correlation and a slightincrease with chlorophyll concentration. Composition of phytoplanktonshowed a dominance of cyanobacteria in hypertrophic lakes, whereasthe importance of chrysophytes and dinophytes decreased withan increase in trophic status. Protozoans (heterotrophic flagellatesand ciliates) made up 24% (mesotrophic lakes) to 42% (hypertrophiclakes) of total zooplankton biomass on average, and were dominatedby ciliates (62–80% of protozoan biomass). Seasonally,protozoans can build up to 60% of zooplankton biomass in spring,when heterotrophic flagellates can contribute     

Relationship between bacteria, phytoplankton and heterotrophic nanoflagellates along the trophic gradient

Bacterial and heterotrophic nanoflagellates (HNF) abundance, as well as bacterial production and chlorophylla levels, were measured at five sites extending from the coastal zone toward the open Adriatic in the period from March to October 1995. The investigated areas were grouped into trophic categories according to concentrations of chlorophylla. All the biotic-para-meters increased along the trophic gradient, leading to eutrophy, but they did not increase at the same rate. The bacterial biomass: phytoplankton biomass (BB: chla) ratio decreased from about 10 in the very oligotrophic area to 0.8 at the eutrophic site. In contrast, the bacterial abundance: HNF abundance ratio (B: HNF) increased from 1000 bacteria per 1 flagellate in the oligotrophic system to 1700 bacteria flagellate⁴ in the eutrophic area. Decreasing BB: chla and increasing B: HNF ratios along the trophic gradient might reflect the different structures of the microbial food web. Relationships between bacterial abundance and production, and chla and HNF showed that bacterial abundance along the trophic gradient was regulated by the interplay between nutrient supply and grazing pressure. But in the oligotrophic system, bacterial abundance was more closely related to bacterial production and chla than in the eutrophic system, suggesting stronger control of bacterial abundance by substrate supply. On the other hand, the coupling between bacteria and HNF, and uncoupling between bacterial abundance and production in the eutrophic system, showed that the importance of bacteriovory increased in richer systems.     

Antimicrobial peptides from marine invertebrates: challenges and perspectives in marine antimicrobial peptide discovery

The emergence of pathogenic bacteria resistance to conventional antibiotics calls for an increased focus on the purification and characterization of antimicrobials with new mechanisms of actions. Antimicrobial peptides are promising candidates, because their initial interaction with microbes is through binding to lipids. The interference with such a fundamental cell structure is assumed to hamper resistance development. In the present review we discuss antimicrobial peptides isolated from marine invertebrates, emphasizing the isolation and activity of these natural antibiotics. The marine environment is relatively poorly explored in terms of potential pharmaceuticals, and it contains a tremendous species diversity which evolved in close proximity to microorganisms. As invertebrates rely purely on innate immunity, including antimicrobial peptides, to combat infectious agents, it is believed that immune effectors from these animals are efficient and rapid inhibitors of microbial growth.     

On the Importance of Planktonic Protozoans in the Eutrophication Process of the Baltic Sea

Analysing the results of various authors recent studies in the pelagic region of the Baltic revealed that protozoan biomass is in the same range or even higher than metazooplankton biomass. The dominant groups of planktonic protozoans are heterotrophic pico- and nanoflagellates (various taxonomic groups), large heterotrophic flagellates (mainly dinoflagellates) and ciliates. Regularly the spring bloom of phytoplankton is accompanied by a maximum of protozoan biomass which declines in early summer as a result of intensive grazing pressure by metazooplankton and changing food conditions. The analysis of results from different stations indicated that biomasses of protozoans increase with an increasing degree of eutrophication. Several trophic levels within the microbial web should be added to the traditional view on the pelagic food web of the Baltic. Our knowledge regarding the quantitative aspect of the microbial matter flux of the Baltic is very limited up to now and complex ecological (and taxonomical) studies using standardized methods including all protozoan components are necessary. Protozoans (various trophic groups and levels), besides bacteria, should be viewed as the metabolically most active heterotrophic component in the pelagic region of the Baltic, their activity should increase with an increasing degree of eutrophication.     

Culturability and Persistence of <Emphasis Type="Italic">Francisella noatunensis</Emphasis> subsp. <Emphasis Type="Italic">orientalis</Emphasis> (syn. <Emphasis Type="Italic">Francisella asiatica</Emphasis>) in Sea- and Freshwater Microcosms

Francisella noatunensis subsp. orientalis (syn. Francisella asiatica), the causative agent of franciselliosis in warm-water fish, is a Gram-negative facultative intracellular bacterium. Although it has been characterized as one of the most pathogenic bacteria in fish, the water conditions that allow for its survival and infectious capacities outside the fish host are not known. Data obtained in this project indicate that both temperature and salinity are important factors in the culturability and persistence of F. noatunensis subsp. orientalis in both sea- and freshwater microcosms. These results indicate that culturable F. noatunensis subsp. orientalis persist for longer periods of time and at higher numbers in seawater, and its persistence is inversely related to water temperature. Moreover, the pathogenic properties of the bacteria suspended in water microcosms appear to decrease after only 24 h and become non-infective after 2 days in the absence of the fish host.     

Migration of N-Alkanes from Diesel Oil along the Bacteria-Ciliates Trophic Chain

The data on the dynamics of the content and qualitative composition of intracellular alkanes in Paramecium caudatum Ehrenberg, 1838 cells feeding on bacteria with different levels of exposure to diesel oil are presented. The possibility of migration of alkanes from diesel oil along the trophic chain from bacteria to ciliates was demonstrated.__________Translated from Izvestiya Akademii Nauk, Seriya Biologicheskaya, No. 3, 2005, pp. 375–379.Original Russian Text Copyright © 2005 by Karetnikova, Zhirkova.     

Repair of damage caused by UV- and X-irradiation in the amoeboflagellateNaegleria gruberi

Summary Cysts ofNaegleria gruberi have a normal UV- and an extremely high X-ray resistance compared to other protozoans. Caffeine and 3-aminobenzamide applied to excysting amoeba after irradiation in the encysted state (UV and X-rays) by feeding with drug-containing bacteria increased lethality, while fractionated irradiation (UV) and liquid-holding (UV and X-rays) increased survival. Illumination with visible light after UV-irradiation restored almost 100% viability. The results are discussed in regard to the activity of repair mechanisms.     

Bacteria-mediated disease therapy

Laboratory bacteria have been model systems for studying gene function or developing molecular biotechnologies; industrial bacteria have been used to produce drugs, foods, fuel etc.; while pathogenic bacteria have been the subject of infectious diseases studies with aim of controlling them. Recently, with our increasing knowledge in molecular biology, bacterial genetics, and immunology, bacteria have gained increased interest in therapeutic applications. This review will summarize recent advances toward applying bacteria in treatment of diseases, such as inflammatory disease, cancer, and virus infection. Probiotics have long been appreciated for their immunomodulatory effect, which are discussed in the anti-inflammatory section. Anaerobic bacteria have been applied for destructing tumor cells or used as a magic bullet to specifically deliver therapeutic agents to tumor tissue. Examples and strategies of applying those tumor-targeting bacteria are discussed in the antitumor section. Finally, the pioneer study of applying Salmonella in delivering RNase P that was specifically engineered to target essential mRNAs of virus is summarized.     

Unpacking brown food‐webs: Animal trophic identity reflects rampant microbivory

Detritivory is the dominant trophic paradigm in most terrestrial, aquatic, and marine ecosystems, yet accurate measurement of consumer trophic position within detrital (=“brown”) food webs has remained unresolved. Measurement of detritivore trophic position is complicated by the fact that detritus is suffused with microbes, creating a detrital complex of living and nonliving biomass. Given that microbes and metazoans are trophic analogues of each other, animals feeding on detrital complexes are ingesting other detritivores (microbes), which should elevate metazoan trophic position and should be rampant within brown food webs. We tested these hypotheses using isotopic (¹⁵N) analyses of amino acids extracted from wild and laboratory‐cultured consumers. Vertebrate (fish) and invertebrate detritivores (beetles and moths) were reared on detritus, with and without microbial colonization. In the field, detritivorous animal specimens were collected and analyzed to compare trophic identities among laboratory‐reared and free‐roaming detritivores. When colonized by bacteria or fungi, the trophic positions of detrital complexes increased significantly over time. The magnitude of trophic inflation was mediated by the extent of microbial consumption of detrital substrates. When detrital complexes were fed to vertebrate and invertebrate animals, the consumers registered similar degrees of trophic inflation, albeit one trophic level higher than their diets. The wild‐collected detritivore fauna in our study exhibited significantly elevated trophic positions. Our findings suggest that the trophic positions of detrital complexes rise predictably as microbes convert nonliving organic matter into living microbial biomass. Animals consuming such detrital complexes exhibit similar trophic inflation, directly attributable to the assimilation of microbe‐derived amino acids. Our data demonstrate that detritivorous microbes elevate metazoan trophic position, suggesting that detritivory among animals is, functionally, omnivory. By quantifying the impacts of microbivory on the trophic positions of detritivorous animals and then tracking how these effects propagate “up” food chains, we reveal the degree to which microbes influence consumer groups within trophic hierarchies. The trophic inflation observed among our field‐collected fauna further suggests that microbial proteins represent an immense contribution to metazoan biomass. Collectively, these findings provide an empirical basis to interpret detritivore trophic identity, and further illuminate the magnitude of microbial contributions to food webs.