Bacterial Aggregations Research Articles

Streptokinase reduces Streptococcus dysgalactiae subsp. equisimilis biofilm formation

BackgroundStreptococcus dysgalactiae subspecies equisimilis (SDSE) is increasingly recognized as an emerging cause of invasive diseases including necrotizing soft tissue infections (NSTIs). In contrast to the closely related Streptococcus pyogenes, SDSE infections mainly affect older and comorbid patients. Biofilm formation has been demonstrated in soft tissue biopsies of S. pyogenes NSTI cases.ResultsHere, we show that bacterial aggregations indicative of biofilms are also present in SDSE NSTI. Although streptokinase (Ska) activity and biofilm formation did not correlate in a diverse set of clinical SDSE isolates, addition of exogenous Ska at an early time point prevented biofilm formation for selected strains. Deletion of ska in SDSE S118 strain resulted in increased biofilm forming capacity. Ska-deficient mutant strain was characterized by a higher metabolic activity and consequent metabolome profiling of biofilms identified higher deposition of a wide range of metabolites as compared to the wild-type.ConclusionsOur results argue that Ska suppresses biofilm formation in SDSE independent of its original plasminogen converting activity. However, the impact of biofilms and its consequences for patient outcomes in streptococcal NSTIs remain to be elucidated.

Design of Multi-Functional Bio-Safe Dental Resin Composites with Mineralization and Anti-Biofilm Properties.

This study aims to develop multi-functional bio-safe dental resin composites with capabilities for mineralization, high in vitro biocompatibility, and anti-biofilm properties. To address this issue, experimental resin composites consisting of UDMA/TEGDMA-based dental resins and low quantities (1.9, 3.8, and 7.7 vol%) of 45S5 bioactive glass (BAG) particles were developed. To evaluate cellular responses of resin composites, MC3T3-E1 cells were (1) exposed to the original composites extracts, (2) cultured directly on the freshly cured resin composites, or (3) cultured on preconditioned composites that have been soaked in deionized water (DI water), a cell culture medium (MEM), or a simple HEPES-containing artificial remineralization promotion (SHARP) solution for 14 days. Cell adhesion, cell viability, and cell differentiation were, respectively, assessed. In addition, the anti-biofilm properties of BAG-loaded resin composites regarding bacterial viability, biofilm thickness, and biofilm morphology, were assessed for the first time. In vitro biological results demonstrated that cell metabolic activity and ALP expression were significantly diminished when subjected to composite extracts or direct contact with the resin composites containing BAG fillers. However, after the preconditioning treatments in MEM and SHARP solutions, the biomimetic calcium phosphate minerals on 7.7 vol% BAG-loaded composites revealed unimpaired or even better cellular processes, including cell adhesion, cell proliferation, and early cell differentiation. Furthermore, resin composites with 1.9, 3.8, and 7.7 vol% BAG could not only reduce cell viability in S. mutans biofilm on the composite surface but also reduce the biofilm thickness and bacterial aggregations. This phenomenon was more evident in BAG7.7 due to the high ionic osmotic pressure and alkaline microenvironment caused by BAG dissolution. This study concludes that multi-functional bio-safe resin composites with mineralization and anti-biofilm properties can be achieved by adding low quantities of BAG into the resin system, which offers promising abilities to mineralize as well as prevent caries without sacrificing biological activity.

Anticorrosion and Antimicrobial Tannic Acid-Functionalized Ti-Metallic Glass Ribbons for Dental Abutment.

In this study, a recently reported Ti-based metallic glass (MG), without any toxic element, but with a significant amount of metalloid (Si-Ge-B, 18 atom %) and minor soft element (Sn, 2 atom %), was produced in ribbon form using conventional single-roller melt-spinning. The produced Ti60Zr20Si8Ge7B3Sn2 ribbons were investigated by differential scanning calorimetry and X-ray diffraction to confirm their amorphous structure, and their corrosion properties were further investigated by open-circuit potential and cyclic polarization tests. The ribbon's surface was functionalized by tannic acid, a natural plant-based polyphenol, to enhance its performance in terms of corrosion prevention and antimicrobial efficacy. These properties can potentially be exploited in the premucosal parts of dental implants (abutments). The Folin and Ciocalteu test was used for the quantification of tannic acid (TA) grafted on the ribbon surface and of its redox activity. Fluorescent microscopy and ζ-potential measurements were used to confirm the presence of TA on the surfaces of the ribbons. The cytocompatibility evaluation (indirect and direct) of TA-functionalized Ti60Zr20Si8Ge7B3Sn2 MG ribbons toward primary human gingival fibroblast demonstrated that no significant differences in cell viability were detected between the functionalized and as-produced (control) MG ribbons. Finally, the antibacterial investigation of TA-functionalized samples against Staphylococcus aureus demonstrated the specimens' antimicrobial properties, shown by scanning electron microscopy images after 24 h, presenting a few single colonies remaining on their surfaces. The thickness of bacterial aggregations (biofilm-like) that were formed on the surface of the as-produced samples reduced from 3.5 to 1.5 μm.

Discovery, structure and function of plasmid mediated shufflon

Antimicrobial resistance has become a major public health issue of global concern. Conjugation is an important way for fast spreading drug-resistant plasmids, during which the type Ⅳ pili plays an important role. Type Ⅳ pili can adhere on the surfaces of host cell and other medium, facilitating formation of bacterial biofilms, bacterial aggregations and microcolonies, and is also a critical factor in liquid conjugation. PilV is an adhesin-type protein found on the tip of type Ⅳ pili encoded by plasmid R64, and can recognize the lipopolysaccharid (LPS) molecules that locate on bacterial membrane. The shufflon is a clustered inversion region that diversifies the PilV protein, which consequently affects the recipient recognition and conjugation frequency in liquid mating. The shufflon was firstly discovered on an IncI1 plasmid R64 and has been identified subsequently in plasmids IncI2, IncK and IncZ, as well as the pathogenicity island of Salmonella typhi. The shufflon consists of four segments including A, B, C, and D, and a specific recombination site named sfx. The shufflon is regulated by its downstream-located recombinase-encoding gene rci, and different rearrangements of the shufflon region in different plasmids were observed. Mobile colistin resistance gene mcr-1, which has attracted substantial attentions recently, is mainly located in IncI2 plasmid. The shufflon may be one of the contributors to fast spread of mcr-1. Herein, we reviewed the discovery, structure, function and prevalence of plasmid mediated shufflon, aiming to provide a theoretical basis on transmission mechanism and control strategy of drug-resistant plasmids.

U (VI) tolerance affects Shewanella sp. RCRI7 biological responses: growth, morphology and bioreduction ability

Native Shewanella sp. RCRI7 is recently counted as an operative bacterium in the uranium bio-reduction. The aim of this study was to investigate the effects of uranium tolerance on the morphology and population of RCRI7, following its potential removal capacity in different time intervals. In this research, the bacterial growth and uranium removal kinetic were evaluated in aerobic TSB medium, uranium-reducing condition (URC), aerobic uranium-containing (AUC) and anaerobic uranium-free (AUF) solution, following evaluations of omcAB gene expressions. In addition, spectrophotometry analyses were performed in URC confirming the bio-reduction mechanism. It was found that the bacteria can grow efficiently in the presence of 0.5mM uranium anaerobically, unlike AUC and AUF solutions. Since the bacterium's adsorption capacity is quickly saturated, it can be deduced that uranium reduction should be dominant as incubation times proceed up to 84h in URC. In 92h incubation, the adsorbed uranium containing unreduced and reduced (U (IV) monomeric), was released to the solution due to either increased pH or bacterial death. In AUC and AUF, improper conditions lead to the reduced bacterial size (coccus-shape formation) and increased bacterial aggregations; however, membrane vesicles produced by the bacteria avoid the uranium incrustation in AUC. In overall, this study implies that Shewanella sp. RCRI7 are well tolerated by uranium under anaerobic conditions and the amount of regenerated uranium increases over time in the reduced form.

Bacterial Invasion of Pulp Blood Vessels in Teeth with Symptomatic Irreversible Pulpitis

IntroductionThis study described the degenerative changes and infection patterns of the pulp tissue associated with symptomatic irreversible pulpitis. MethodsThe material consisted of 32 extracted teeth with untreated deep caries that were clinically and histologically diagnosed with irreversible pulpitis and were part of the histopathologic collection of 1 of the authors. The controls consisted of intact teeth with normal uninflamed pulps and teeth with reversible pulpitis. Teeth were processed for histopathologic and histobacteriologic analyses. ResultsAll teeth with irreversible pulpitis showed areas of severe acute inflammation, necrosis, microabscesses, and bacterial infection in the pulp chamber. These areas were surrounded by a chronic inflammatory infiltrate, and, at the distance, the pulp tissue was often uninflamed. Bacteria were also observed in the areas surrounding the necrotic foci, both as scattered cells through the extravascular space and at varying numbers within the blood vessel lumen. The number of bacteria and the density of the intravascular bacterial aggregations varied considerably. In one third of the cases, bacteria occurred in the lumen of venules in areas at a considerable distance from the necrotic focus in the coronal third of the root. No intravascular bacteria were noted in the middle and apical segments of the canal. No bacteria were found in the pulps of any of the control specimens. ConclusionsBacterial invasion and colonization of necrotic areas were observed in the pulp of all teeth with caries exposure and symptomatic irreversible pulpitis. Bacterial penetration of blood vessels occurred in all cases, suggesting that this may be an important mechanism of spread of bacterial infection through the pulp tissue in an endodontic infection.

Pulp Response to Periodontal Disease: Novel Observations Help Clarify the Processes of Tissue Breakdown and Infection

IntroductionThis study evaluated the pulp response to periodontal disease of increasing severity. MethodsThe material comprised human teeth affected by moderate (n = 16) to severe (n = 48) periodontal disease and no clinically identified caries lesions. Specimens were obtained by extraction and were processed for histopathologic and histobacteriologic methods. ResultsIn 13 of 16 teeth with moderate periodontal disease and vital pulp, no frank accumulations of inflammatory cells were observed. In 22 of 32 teeth with severe periodontal disease and vital pulps, no distinct inflammatory cell accumulations were observed in any portion of the pulp when there was an intact or minimally damaged cementum layer in the corresponding areas. Intravascular bacterial aggregations were detected in pulp blood vessels in 6 teeth with symptomatic pulpitis and severe periodontal disease, which had not reached the root apex in 4 of them. Focal areas of infection and varying accumulations of acute and chronic inflammatory cells were observed throughout the pulp tissue and surrounding these infected blood vessels. When the periodontal pocket reached the apex of single-rooted teeth, the entire canal space was necrotic and colonized by bacteria, often forming biofilms. In multirooted teeth, the pulp response varied according to the root(s) affected. Intrapulpal calcifications were observed in 91% of the teeth. The pulp and cementum of the control teeth showed no pathologic changes or bacteria. ConclusionsThe pulp showed a significantly detectable response when the cementum coverage was lost or when the periodontal pocket reached the root apex. In the former condition, the pulp response was usually discrete, whereas in the latter, severe reactions usually developed. In some teeth, vessels with a compromised blood flow may serve as avenues for bacteria to invade the pulp via apical or lateral foramina. This indicates that in some teeth the pulp may undergo severe inflammation and necrosis even before the periodontal disease reaches the apical root segment.

Bacteriologic status of non-cavitated proximal enamel caries lesions. A histologic and histobacteriologic study

ObjectivesThis histologic and histobacteriologic study evaluated non-cavitated interproximal white spot lesions in human teeth for the 1) presence and morphology of bacterial aggregations at the enamel-dentinal junction (EDJ), and 2) reactions in the subjacent pulp tissue. MethodsThe material comprised 16 third molars diagnosed with early interproximal caries lesions obtained consecutively in a single clinical practice. Four third molars with clinically intact proximal surfaces served as controls. Caries activity was categorized as active or arrested. Teeth were prepared for histologic and histobacteriologic analyses. ResultsControl teeth exhibited normal tissue conditions with no bacteria. Macroscopic discoloration of the superficial dentin occurred in 14/16 teeth with early caries lesions. Bacterial aggregations resembling biofilms were observed in 10/16 teeth (six with active and four with arrested lesions). Bacterial cells were observed superficially in some of the underlying dentinal tubules, while in a few cases a deeper penetration was evident. Pathologic changes of varying degrees were observed in the pulps of all 16 teeth, regardless of the caries activity (active or arrested). Pulp reactions varied from disruption of the odontoblast layer, with loss of odontoblasts, to formation of tertiary dentin. ConclusionsBacterial biofilms associated with white-spot caries lesions may traverse the enamel and reach the underlying dentin in both active and arrested lesions. In all teeth with early lesions, the pulps showed changes in response to the very superficial biofilm challenge. Clinical significanceIn non-cavitated, active or arrested, enamel caries lesions, bacteria traverse the enamel and may establish structured biofilms at the enamel-dentinal junction, causing early pulp changes. These new findings may stimulate clinicians to rethink the rationale for treatment methods that are based on the assumption that bacteria are absent in white-spot lesions.

Generating Controlled, Dynamic Chemical Landscapes to Study Microbial Behavior.

We demonstrate a method for the generation of controlled, dynamic chemical pulses-where localized chemoattractant becomes suddenly available at the microscale-to create micro-environments for microbial chemotaxis experiments. To create chemical pulses, we developed a system to introduce amino acid sources near-instantaneously by photolysis of caged amino acids within a polydimethylsiloxane (PDMS) microfluidic chamber containing a bacterial suspension. We applied this method to the chemotactic bacterium, Vibrio ordalii, which can actively climb these dynamic chemical gradients while being tracked by video microscopy. Amino acids, rendered biologically inert ('caged') by chemical modification with a photoremovable protecting group, are uniformly present in the suspension but not available for consumption until their sudden release, which occurs at user-defined points in time and space by means of a near-UV-A focused LED beam. The number of molecules released in the pulse can be determined by a calibration relationship between exposure time and uncaging fraction, where the absorption spectrum after photolysis is characterized by using UV-Vis spectroscopy. A nanoporous polycarbonate (PCTE) membrane can be integrated into the microfluidic device to allow the continuous removal by flow of the uncaged compounds and the spent media. A strong, irreversible bond between the PCTE membrane and the PDMS microfluidic structure is achieved by coating the membrane with a solution of 3-aminopropyltriethoxysilane (APTES) followed by plasma activation of the surfaces to be bonded. A computer-controlled system can generate user-defined sequences of pulses at different locations and with different intensities, so as to create resource landscapes with prescribed spatial and temporal variability. In each chemical landscape, the dynamics of bacterial movement at the individual scale and their accumulation at the population level can be obtained, thereby allowing the quantification of chemotactic performance and its effects on bacterial aggregations in ecologically relevant environments.

Bacteria push the limits of chemotactic precision to navigate dynamic chemical gradients

Ephemeral aggregations of bacteria are ubiquitous in the environment, where they serve as hotbeds of metabolic activity, nutrient cycling, and horizontal gene transfer. In many cases, these regions of high bacterial concentration are thought to form when motile cells use chemotaxis to navigate to chemical hotspots. However, what governs the dynamics of bacterial aggregations is unclear. Here, we use an experimental platform to create realistic submillimeter-scale nutrient pulses with controlled nutrient concentrations. By combining experiments, mathematical theory, and agent-based simulations, we show that individual Vibrio ordalii bacteria begin chemotaxis toward hotspots of dissolved organic matter (DOM) when the magnitude of the chemical gradient rises sufficiently far above the sensory noise that is generated by stochastic encounters with chemoattractant molecules. Each DOM hotspot is surrounded by a dynamic ring of chemotaxing cells, which congregate in regions of high DOM concentration before dispersing as DOM diffuses and gradients become too noisy for cells to respond to. We demonstrate that V. ordalii operates close to the theoretical limits on chemotactic precision. Numerical simulations of chemotactic bacteria, in which molecule counting noise is explicitly taken into account, point at a tradeoff between nutrient acquisition and the cost of chemotactic precision. More generally, our results illustrate how limits on sensory precision can be used to understand the location, spatial extent, and lifespan of bacterial behavioral responses in ecologically relevant environments.

Denitrifying granules in a marine Upflow Anoxic Sludge Bed (UASB) reactor

Marine land-based Recirculating Aquaculture Systems (RAS) are generally perceived as environmentally friendly aquatic production systems. To promote their sustainability even further and reduce the discharge of nutrients, there is a need for cost-effective end-of-pipe treatment technologies for removing nutrients. This includes nitrate-nitrogen (NO3−-N) for which well-proven technologies for freshwater systems exists, while similar technologies for saltwater systems are less advanced. Granular technology has been developed since the 1970s in wastewater treatment under the upflow anaerobic sludge bed (UASB) concept. This concept is based on the enrichment of different bacterial aggregations into a compact granule, optimizing synergistic and syntrophic bacterial processes by reducing the diffusion distance of substrates between the different bacterial consortia forming the granule. The following study examined the: 1) granular formation; and 2) nitrate removal capacity of a marine Upflow Anoxic Sludge Bed (UASB) reactor operating at different up-flow velocities (0.40–2.11 m/h). The results showed that marine denitrifying granules developed within 27 days using preconditioned rainbow trout (Oncorhynchus mykiss) organic matter waste, and that the highest specific denitrification rate (321.9 ± 13.1 mg NO3−-N/g Total Volatile Suspended Solids (TVSS)/d) was found at an upflow velocity of 0.97 m/h. The marine UASB denitrifying granule reactor had a total capacity of removing 14.9 kg NO3−-N/m3 reactor volume per day at a hydraulic retention time of 1.9 h, making it a strong candidate for end-of-pipe denitrification of marine RAS effluent as well as for in-line treatment in marine systems.

Evaluation of chromocystoscopy in the diagnosis of cystitis in female donkeys.

Early detection of cystitis in equine is essential to improve the prognosis and outcome of therapy. However, the conventional white light endoscopy is not sufficiently accurate for this purpose. Hence, this study evaluated chromoendoscopy as a recent diagnostic tool for cystitis in female donkeys. For this purpose, 5 apparently normal donkeys (control group) and 5 female donkeys with cystitis (diseased group) were used. Physical and rectal examinations, urine analysis, white light cystoscopy, methylene blue-based chromoendoscopy and histopathology were performed in all animals. Turbid urine exhibiting an alkaline pH and a significant (P = .02) increase in the numbers of RBCs and WBCs was observed in the diseased group compared to the control one. In the control group, white light cystoscopy showed a smooth pale pink glistening mucosa with two openings of the ureters and visible submucosal blood vessels. During chromocystoscopy, faint bluish discoloration of the mucosal surface with clearly visible submucosal blood vessels was detectable. These findings were correlated with the histopathological findings of the biopsies collected from the urinary bladder. In the diseased group, white-light cystoscopy showed clearly visible blood vessels, mildly hyperaemic mucosa in focal or diffuse forms and small vesicle formation. Chromocystoscopy revealed dark bluish oedematous and irregular mucosa either in a focal form or a diffuse form (marbled appearance) with deeply stained submucosal blood vessels. Histopathologically, the urothelium was hyperplastic with squamous metaplasia and the lamina propria was infiltrated with few leukocytes and congested blood vessels. Small bluish dots representing the absorbed methylene blue dye were seen in the inflamed areas against the lightly stained mucosa of the bladder. Severe diffuse necrotic cystitis was also seen with bacterial aggregations on the surface. Gram’s staining revealed both gram positive bacilli and Gram positive coccobacilli. In conclusion, chromoendoscopy is a helpful tool for early diagnosis of cystitis in female donkeys and enables targeted biopsies, which improves the prognosis and outcome of therapy.

Assessing the effect of different shapes of glyco-gold nanoparticles on bacterial adhesion and infections.

Achieving selective and sensitive carbohydrate-protein interactions (CPIs) using nanotechnology is an intriguing area of research. Here we demonstrate that the different shapes of gold nanoparticles (AuNPs) functionalized with monosaccharides tune the bacterial aggregations. The mechanism of aggregation revealed that the large number of surface interactions of rod shaped mannose-AuNPs with E. coli ORN 178 compared with spherical and star-shaped AuNPs exhibited higher avidity and sensitivity. Moreover, such sensitive binding can be used for effective inhibition of bacterial infection of cells.

Extraradicular Infection as the Cause of Persistent Symptoms: A Case Series

IntroductionThis article describes 3 cases that presented persistent symptoms after appropriate endodontic treatment. Histopathologic and histobacteriologic investigation were conducted for determination of the cause. MethodsThree cases are reported that presented with persistent symptoms after endodontic retreatment (cases 1 and 2) or treatment (case 3). Periapical surgery was indicated and performed in these cases. The biopsy specimens, consisting of root apices and the apical periodontitis lesions, were subjected to histopathologic and histobacteriologic analyses. ResultsCase 1 was an apical cyst with necrotic debris, heavily colonized by ramifying bacteria, in the lumen. No bacteria were found in the apical root canal system. Case 2 was a granuloma displaying numerous bacterial aggregations through the inflammatory tissue. Infection was also present in the dentinal tubules at the apical root canal. Case 3 was a cyst with bacterial colonies floating in its lumen; bacterial biofilms were also seen on the external apical root surface, filling a large lateral canal and other apical ramifications, and between layers of cementum detached from the root surface. No bacteria were detected in the main root canal. ConclusionsDifferent forms of extraradicular infection were associated with symptoms in these cases, leading to short-term endodontic failure only solved by periapical surgery.

Computer 3D controlled bacterial transports and aggregations of microbial adhered nano-components

Bacteria can be used as microcarriers for the transport of nano-components. By exploiting specific taxes such as chemotaxis, aerotaxis or phototaxis, to name but a few examples, sensory-based transports of nano-components towards or away from a specific source of chemical, oxygen, or light for instance, can be achieved. Using microbial adhesion methods, such nano-components can be attached to the surface of magnetotactic bacteria (MTB), allowing for precise magnetotaxis computer controlled transport of these nano-components along a planned path on a planar surface or towards a specific region within a 3D space. Here, the possibility of attaching a large number of nano-components to the surface of magnetotactic bacteria combined with the possibility of aggregating a large number of cells in a 3D space suggests the possibility of delivering and aggregating a large number of nano-components at a specific region within a relatively large 3D space using relatively low electrical energy.

Physicochemical parameters aid microbial community? A case study from marine recreational beaches, Southern India

A total of 176 (water and sediment) samples from 22 stations belonging to four different (urban, semi-urban, rural, and holy places) human habitations of Tamil Nadu beaches were collected and analyzed for physiochemical and microbial parameters during 2008-2009. Bacterial counts were two- to tenfold higher in sediments than in water due to strong bacterial aggregations by dynamic flocculation and rich organic content. The elevated bacterial communities during the monsoon explain rainfalls and several other wastes from inlands. Coliform counts drastically increased at holy and urban places due to pilgrimage and other ritual activities. Higher values of the pollution index (PI) ratio (>1) reveals, human fecal pollutions affect the water quality. The averaged PI ratio shows a substantial higher microbial contamination in holy places than in urban areas and the order of decreasing PI ratios observed were: holy places > urban areas > semi-urban areas > rural areas. Correlation and factor analysis proves microbial communities were not related to physicochemical parameters. Principal component analysis indicates 55.32% of the total variance resulted from human/animal fecal matters and sewage contaminants whereas 19.95% were related to organic contents and waste materials from the rivers. More than 80% of the samples showed a higher fecal coliform and Streptococci by crossing the World Health Organization's permissible limits.

Air–liquid interface biofilm formation by psychrotrophic pseudomonads recovered from spoilt meat

The ability to colonise the surface of liquids has obvious advantages for bacteria and biofilm formation at the meniscus and air-liquid (A-L) interface is common amongst environmental pseudomonads. Bacteria from this genus also colonise raw meat and in this work the ability of these to produce biofilms was assessed. Sixty isolates were recovered from vacuum-packed venison, phenotypically characterised and shown by hierarchical cluster analysis to represent a diverse collection of psychrotrophic spoilt venison-associated pseudomonads. Of these, 12% were found to produce biofilms limited to the meniscus region of the microcosm walls, 31% produced attached biofilms with significant extensions across the A-L interface and 45% produced unattached 'floating' biofilms. A combined statistical analysis of growth, biofilm strength and attachment levels revealed that growth affected strength but not attachment, and that there was a significant relationship between attachment and strength. Some environmental pseudomonads are known to utilise cellulose as a biofilm matrix component and here 28% of the SVP isolates were found to express cellulose by epifluorescent microscopy. This survey suggests that biofilm formation may be more common in psychrotrophic meat-associated isolates than amongst the wider pseudomonad community from which spoilage bacteria might be recruited. This may reflect a selective advantage of bacterial aggregations such as biofilms in environments subject to high levels of physical disturbance. Aggregations may be more resistant to competition and dehydration stress than individual bacteria, whilst fragments of these aggregations may prove more effective in the colonisation of new habitats.

Biofilms-The Unforgiving Film in Dentistry (Clinical Endodontic Biofilms)

It is evident that oral microorganisms have the capacity to respond and adapt to changing environmental conditions. Individual microorganisms are able to sense and process the chemical information from the environment and thereby adjust their phenotypic properties. The term biofilm is used to indicate the presence of a film of condensed microorganisms on any surface. Bacterial condensations on the walls of infected root canals have been observed suggesting that mechanisms for biofilm formation also exist inside the root canal space. A mature biofilm is a metabolically active community of microorganisms where individuals share duties and benefits. A growing body of knowledge suggests that organisms in biofilms assume a stronger pathogenic potential than those in a planktonic state. From these aspects, the formation of biofilms carries particular clinical significance because not only host defense mechanisms, but also therapeutic efforts including chemical and mechanical anti-microbial treatment measures, have a most difficult task to deal with organisms that are gathered in a biofilm. Such bacterial aggregations have been thought to be the cause of therapy-resistant apical periodontitis. The aim of this communication is therefore to give an overview of the biofilm concept and to discuss how it may apply to endodontic infections.

Strange Tadpoles from the Lower Miocene of Turkey: Is Paedogenesis Possible in Anurans?

Fossil material from the lower Miocene collected in the basin lake of Beskonak (Turkey) included 19 slabs showing 19 amphibian anuran tadpoles of rather large size, at Gosner stages 36–38. These well preserved specimens show many morphological and skeletal characters. They are here tentatively referred to the genus Pelobates. Two of these tadpoles show an unusual group of black roundish spots in the abdominal region, and a third similar group of spots is present in another slab but we were unable to state if it was associated with a tadpole or not. Several hypotheses can be proposed to account for these structures: artefacts; intestinal content (seeds; inert, bacterial or fungal aggregations; eggs); internal or external parasites; diseases; eggs produced by the tadpole. The latter hypothesis is discussed in detail and is shown to be unlikely for several reasons. However, in the improbable case where these spots would correspond to eggs, this would be the first reported case of natural paedogenesis in anurans, a phenomenon which has been so far considered impossible mostly for anatomical reasons (e.g., absence of space in the abdominal cavity).

Asymmetry in the Expression of Bilateral Morphs as an Indicator Showing the Character of Interrelations Between Fleas and the Plague Pathogen

The level of fluctuating asymmetry (FA) of bilateral signs in fleas was shown to reflect a certain character of their interrelations with Yersinia pestis. Flea imagoes that had bacterial aggregations in their proventriculi demonstrated greater signs of FA chaetotaxy. Besides, in the experiments with groups of fleas originally differing in the intensity of FA expression, increased vector capacity was characteristic of individual fleas with higher levels of this index or those caught in the areas of the natural foci where epizootics had been registered. These peculiar interrelations manifested themselves both at intra- and inter-population levels in several species of Siphonaptera order, as well as in the imago hybrids originating from crosses between C.tesquorum altaicus and C.t.sungaris.