Application of the galactomannan gel from Cassia grandis seeds for biomedical purposes: Study of the incorporation of collagenases and their release profile.

The galactomannan-based gel from Cassia grandis seeds was used to incorporate Penicillium sp. UCP 1286 and commercial collagenases. Experiments were carried out according to a 23-full factorial design to identify the most significant parameters for the incorporation process. The pH of the incorporation solution (pHi), stirring time (t), and initial protein concentration in the crude extract (PCi) were selected as the three independent variables, and the efficiency of collagenase incorporation (E) and collagenolytic activity (CA) after 360 min as the responses. pHi and PCi showed positive statistically significant effects on E, while CA was positively influenced by pHi and t, but negatively by PCi. The fungi collagenase was released from the gel following a pseudo-Fickian behavior. Additionally, no <76 % of collagenase was efficiently incorporated into the gel retaining a high CA (32.5-69.8 U/mL). The obtained results for the commercial collagenase (E = 93.88 %, CA = 65.8 U/mL, and n = 0.10) demonstrated a pseudo-Fickian behavior similar to the fungi-collagenase. The results confirm the biotechnological potential of the gel as an efficient matrix for the incorporation of catalytic compounds; additionally, the incorporation of collagenases was achieved by retaining the proteases CA and releasing them in a controlled manner.

Drug Extended-Release System Utilizing Micelle Formation of Highly Water-Soluble Drugs and a Counter Polymer.

The objective of this study is to clarify the mechanism of extending release of highly water-soluble drugs via counter polymer (CP) utilization in poly(ethylene oxide) (PEO)/polyethylene glycol (PEG) matrix tablets. Carbomer, poly(acrylic acid), was used as a CP, which has the opposite charges to the drugs. The in vitro release of several highly water-soluble drugs from PEO/PEG tablet with or without CP were tested, the relationship between the sustained release effect by a CP (SRE) and the physicochemical properties of the drugs was investigated. The results demonstrated that the utilization of CP can extend the release of some highly water-soluble drugs by effectively controlling the drug diffusion through matrices. On the other hand, the effectiveness of CP was different depending on the drugs applied. There were not statistical correlations between SRE and physicochemical properties such as solubility, molecular weight, and charge intensity of the drugs, while a micelle forming property of the drugs played an important role in SRE by CP. It was concluded that CP, Carbomer, having negative charges could effectively interact with opposite charges on the surface of stable drug micelles, which could result in a significant decrease in drug diffusion leading to extended drug release. It is considered that the system utilizing CP is a promising approach to achieve extended release of highly water-soluble drugs with a reasonable tablet size, especially in the case of large drug loading.

Luminescent Hybrid BPA.DA-NVP@Eu2L3 Materials: In Situ Synthesis, Spectroscopic, Thermal, and Mechanical Characterization.

A series of homogeneous hybrid BPA.DA-NVP@Eu2L3 materials were obtained through an in situ approach where the luminescent dopant was formed at the molecular level with different contents (0.1; 0.2; 0.5; 1; and 2% by weight). A Europium(III) complex (Eu2L3) with quinoline-2,4-dicarboxylic acid was applied as a luminescence additive while a polymer matrix consisted of a combination of bisphenol A diacrylate (BPA.DA) and N-vinylpyrrolidone (NVP) monomers. Synthesis steps and the final materials were monitored by NMR and Fourier transform infrared spectroscopy (FTIR). The emission, excitation spectra, lifetime, and quantum yield measurements were applied for the determination of the photophysical characteristics. The thermal and mechanical properties of the obtained materials were tested via thermal analysis methods (TG/DTG/DSC and TG-FTIR) in air and nitrogen atmospheres, dynamic mechanical analysis (DMA), and hardness and bending measurements. Generally, even a small addition of the metal complex component causes changes in the thermal, mechanical, and luminescent properties. Hybrid materials with a greater europium complex content are characterized by a lower stiffness and hardness while the heterogeneity and the flexibility of the samples increase. A very small amount of an Eu2L3 admixture (0.1% wt.) in a hybrid material causes an emission in the red spectral range and the luminescence intensity was reached for the BPA-DA-NVP@1%Eu2L3 material. These materials may be potentially used in chemical sensing, security systems, and protective coatings against UV.

Encapsulation of Methanotrophs within a Polymeric Matrix Containing Copper- and Iron-Based Nanoparticles to Enhance Methanol Production from a Simulated Biogas.

The production of renewable energy or biochemicals is gaining more attention to minimize the emissions of greenhouse gases such as methane (CH4) and carbon dioxide for sustainable development. In the present study, the influence of copper (Cu)- and iron (Fe)-based nanoparticles (NPs), such as Cu, Fe3O4, and CuFe2O4, was evaluated during the growth of methanotrophs for inoculum preparation and on the development of a polymeric-matrix-based encapsulation system to enhance methanol production from simulated biogas (CH4 and CO2). The use of simulated biogas feed and the presence of NP-derived inoculums produce a remarkable enhancement in methanol production up to 149% and 167% for Methyloferula stellata and Methylocystis bryophila free-cells-based bioconversion, respectively, compared with the use of pure CH4 as a control feed during the growth stage. Furthermore, these methanotrophs encapsulated within a polymeric matrix and NPs-based systems exhibited high methanol production of up to 156%, with a maximum methanol accumulation of 12.8 mmol/L over free cells. Furthermore, after encapsulation, the methanotrophs improved the stability of residual methanol production and retained up to 62.5-fold higher production potential than free cells under repeated batch reusability of 10 cycles. In the presence of CH4 vectors, methanol production by M. bryophila improved up to 16.4 mmol/L and retained 20% higher recycling stability for methanol production in paraffin oil. These findings suggest that Cu and Fe NPs can be beneficially employed with a polymeric matrix to encapsulate methanotrophs and improve methanol production.

Poly(butylene adipate-co-terephthalate)/Poly(lactic acid) Polymeric Blends Electrospun with TiO2-R/Fe3O4 for Pollutant Photodegradation.

This work aimed to use the electrospinning technique to obtain PBAT/PLA polymer fibers, with the semiconductors rutile titanium dioxide (TiO2-R) and magnetite iron oxide (Fe3O4), in order to promote the photocatalytic degradation of environmental contaminants. The parameters used in the electrospinning process to obtain the fibers were distance from the needle to the collecting target of 12 cm, flow of 1 mL h-1 and voltage of 14 kV. The best mass ratio of semiconductors in the polymeric fiber was defined from a 22 experimental design, and the values obtained were 10% TiO2-R, 1% Fe3O4 at pH 7.0. Polymer fibers were characterized by Scanning Electron Microscopy (SEM), Differential Scanning Calorimetry (DSC), X-ray Diffraction (XRD), Thermogravimetric Analysis (TGA) and Fourier Transform Infrared (FTIR) techniques. SEM measurements indicated a reduction in fiber diameter after the incorporation of semiconductors; for the PBAT/PLA fiber, the average diameter was 0.9466 ± 0.2490 µm, and for the fiber with TiO2-R and Fe3O4 was 0.6706 ± 0.1447 µm. In the DSC, DRX, TGA and FTIR analyses, it was possible to identify the presence of TiO2-R and Fe3O4 in the fibers, as well as their interactions with polymers, demonstrating changes in the crystallinity and degradation temperature of the material. These fibers were tested against Reactive Red 195 dye, showing an efficiency of 64.0% within 24 h, showing promise for photocatalytic degradation of environmental contaminants.

One-pot facile approach to design an efficient macro-porous polymeric matrix to remediate Hg(II)and Pb(II) from aqueous medium and its performance evaluation study by mathematical modelling.

In the present scenario discharge of heavy-metal ions into water bodies is a global threat that is causing serious health hazards even in low concentrations. Thus, in order to remediate the heavy-metal [Hg(II) and Pb(II)] toxicity, an organic-inorganic hybrid functional porous metallo-polymeric network i. e, poly(Zirconyl methacrylate-co-1-vinyl imidazole) (pZrVIm) was fabricated via one-pot facile synthesis approach. The pZrVIm architecture has shown high removal efficiency for Hg(II) and Pb(II) aqueous medium even in extremely low quantities. Advanced instrumental techniques were used to characterize the structural and morphological characteristics of pZrVIm. Different experimental variables i.e., reaction time, pH, initial feed concentration, co-ion effects etc. were explored to examine adsorption behaviour. The maximum adsorption capacities (qmax) of pZrVIm5 were calculated as 168.06 and 162.34 mg g-1 for Hg(II) and Pb(II) respectively by the Langmuir isotherm model. Data from isotherms showed that monolayer adsorption on a homogeneous surface is the rate-limiting stage and followed pseudo-second-order kinetic process. The Artificial Neural Network (ANN) modelling was used to validate kinetics and isotherm data which revealed high accuracy of the model with correlation coefficient values (R = 0.99). Various types of isotherm models such as Langmuir, Freundlich, Dubinin-Radushkevich, Temkin, Redlich-Peterson, Toth and Koble-Corigen have been studied to determine the adsorption phenomena. The pore diffusion model revealed breakthrough time of 91 h and 84 h, Hg(II) and Pb(II) with the feed concentration of 15 mg L-1 respectively. The study revealed that pZrVIm5 has great potential for heavy metal ions remediation for water treatment.

A Selective Review on the Novel Approaches and Potential Control Agents of Anti-biofouling and Anti-biofilming.

Specific aggregates of bacterial colonies, which, when embedded in a self-produced matrix, are known to cause biofouling. These complex structures are highly resistant to extreme stress conditions like antibiotics, complex cleaning strategies, or even the human immune system. The formation of biofilm matrices and biofouling is chosen as the preferred microbial environment owing to the advantages offered for the embedded cells for their long-term survival. This increased resistance and virulence emphasizes the urgency of finding newer alternatives for its complete eradication. Combined approaches using enzymes, biomimetic surface modification, and a fusion of physical and chemical methods are gaining more prominence. In brief, this review discusses the structure and biology of biofilms, their mechanism of action, and the new-age anti-biofilm and biofouling agents in food industries, bioelectric, and medical devices. Current anti-biofouling methods, including engineered polymers, surface coatings to antimicrobials, and antibiotics, have also been described.

Toward Thermochromic VO2 Nanoparticles Polymer Films Based Smart Windows Designed for Tropical Climates.

Thermochromic smart windows have been extensively investigated due to two main benefits: first, the comfort for people in a room through avoiding high temperatures resulting from solar heating while taking advantage of the visible light, and second, the energy efficiency saving offered by using those systems. Vanadium dioxide (VO2) is one of the most used materials in the development of thermochromic devices. The countries located in the tropics show little use of these technologies, although studies indicate that due to their characteristics of solar illumination and temperature, they could benefit greatly. To optimize and achieve maximum benefit, it is necessary to design a window that adjusts to tropical conditions and at the same time remains affordable for extensive implementation. VO2 nanoparticles embedded in polymeric matrices are an option, but improvements are required by means of studying different particle sizes, dopants and polymeric matrices. The purpose of this review is to analyze what has been regarding toward the fabrication of smart windows based on VO2 embedded in polymeric matrices for tropical areas and provide a proposal for what this device must comply with to contribute to these specific climatic needs.

Recent advancements on hyperthermia driven controlled drug delivery from nanotherapeutics.

Previous reviews of the works on magnetic nanoparticles for hyperthermia induced treatment concentrated mostly on magnetic fluid hyperthermia (MFH) employing monometallic/metal oxide nanocomposites. In the literature, the word "hyperthermia" was also limited to the use of heat for medicinal purposes. A number of publications have recently been published demonstrating that magnetic nanoparticle-based hyperthermia may produce restricted high temperatures, resulting in the release of medicines that are either connected to the magnetic nanoparticles or encased in polymer matrices. In this debate, we propose broadening the concept of "hyperthermia" to encompass temperature-based treatment as well as magnetically controlled medication delivery. The review also addresses core-shell magnetic nanomaterials, particularly nanoshells made by stacked assembly, for the use of hyperthermia-based treatment and precise administration of drugs. The primary objective of this review article is to demonstrate how the combination of hyperthermia-induced therapy and 'on demand' drug release models may lead to effective applications in personalized medicine.

Numerical Study of Drift Influence on Diffusion Transport through the Hybrid Membrane.

Sodium alginate membranes filled with iron oxide nanoparticles consist of a mixture of organic and inorganic phases. This design offers the possibility to combine the polymer's easy processability and superior separation performance. For a better understanding of the mechanisms of mixture separation, we analyze the diffusion motion of a particle in the hybrid membrane environment. We model structures of two-dimensional heterogenic membranes, which resemble real membrane structures, and then we simulate a random walk on them. We investigate how the additional action of drift changes the motion properties of the diffusing particles through the polymeric membrane filled with inorganic powder. We test the effect of two parameters: the distribution of obstacles (filling) in the membrane and the value of drift on the nature of diffusion. It appears that the synergy between drift, the diffusion, and the membrane structure affect the occurrence of the superdiffusive and subdiffusive character of particle motion as measured by the time-averaged mean square displacement. An important point is the observation that the strong drift supports subdiffusive motion as it increases the chances of particle trapping. Moreover, there exists the optimal value of drift, for which the transport through a membrane speeds up and does not cause trapping.

Preparation and Characterization of Hydrophilic Polymer Based Sustained-Release Matrix Tablets of a High Dose Hydrophobic Drug.

The objective of this study was the preparation and characterization of a sustained-release matrix tablet containing a high-dose hydrophobic drug and its comparison with marketed products. In the present study, HPMC was applied as the matrix-forming polymer for the sustained release of clarithromycin (500 mg). The compatibility of clarithromycin and excipients was studied using a binary mixture approach and compatible excipients were selected. Matrix tablets were prepared using the high-shear wet granulation technique. Tablets were compressed using oblong (19 mm), shallow concave punches, under a compression weight of 900 mg/tablet. The flow of granules was evaluated by determining their bulk density, tapped density, angle of repose, Hausner ratio, and Car's index. Compressed tablets were tested for their physical parameters, mechanical characteristics, drug content, and in vitro drug release, as per United States Pharmacopeia (USP). Clarithromycin is a drug having poor water solubility and showed compatibility with all the excipients used in the formulation of polymeric matrix tablets. FTIR spectra of clarithromycin, before and after being subjected to the stress conditions, confirmed the compatibility of clarithromycin and other ingredients of the matrix tablets. All the formulations exhibited good rheological characteristics and all the parameters related to flow showed results in the acceptable range. Physically, matrix tablets were smooth and shiny, without any surface defects. Weight variation (±5%) and drug content of the tablets (95-102%) were within the pharmacopeial limits. HPMC successfully sustained the drug release for 24 h. It is concluded from the study that dissolution rate of clarithromycin can be sustained using hydrophilic polymer (HPMC) as a release-controlling agent.

The Effects of Three-Dimensional Ligand Immobilization on Kinetic Measurements in Biosensors.

The field of biosensing is in constant evolution, propelled by the need for sensitive, reliable platforms that provide consistent results, especially in the drug development industry, where small molecule characterization is of uttermost relevance. Kinetic characterization of small biochemicals is particularly challenging, and has required sensor developers to find solutions to compensate for the lack of sensitivity of their instruments. In this regard, surface chemistry plays a crucial role. The ligands need to be efficiently immobilized on the sensor surface, and probe distribution, maintenance of their native structure and efficient diffusion of the analyte to the surface need to be optimized. In order to enhance the signal generated by low molecular weight targets, surface plasmon resonance sensors utilize a high density of probes on the surface by employing a thick dextran matrix, resulting in a three-dimensional, multilayer distribution of molecules. Despite increasing the binding signal, this method can generate artifacts, due to the diffusion dependence of surface binding, affecting the accuracy of measured affinity constants. On the other hand, when working with planar surface chemistries, an incredibly high sensitivity is required for low molecular weight analytes, and furthermore the standard method for immobilizing single layers of molecules based on self-assembled monolayers (SAM) of epoxysilane has been demonstrated to promote protein denaturation, thus being far from ideal. Here, we will give a concise overview of the impact of tridimensional immobilization of ligands on label-free biosensors, mostly focusing on the effect of diffusion on binding affinity constants measurements. We will comment on how multilayering of probes is certainly useful in terms of increasing the sensitivity of the sensor, but can cause steric hindrance, mass transport and other diffusion effects. On the other hand, probe monolayers on epoxysilane chemistries do not undergo diffusion effect but rather other artifacts can occur due to probe distortion. Finally, a combination of tridimensional polymeric chemistry and probe monolayer is presented and reviewed, showing advantages and disadvantages over the other two approaches.

Immobilized enzymes and cell systems: an approach to the removal of phenol and the challenges to incorporate nanoparticle-based technology.

The presence of phenol in wastewater poses a risk to ecosystems and human health. The traditional processes to remove phenol from wastewater, although effective, have several drawbacks. The best alternative is the application of ecological biotechnology tools since they involve biological systems (enzymes and microorganisms) with moderate economic and environmental impact. However, these systems have a high sensitivity to environmental factors and high substrate concentrations that reduce their effectiveness in phenol removal. This can be overcome by immobilization-based technology to increase the performance of enzymes and bacteria. A key component to ensure successful immobilization is the material (polymeric matrices) used as support for the biological system. In addition, by incorporating magnetic nanoparticles into conventional immobilized systems, a low-cost process is achieved but, most importantly, the magnetically immobilized system can be recovered, recycled, and reused. In this review, we study the existing alternatives for treating wastewater with phenol, from physical and chemical to biological techniques. The latter focus on the immobilization of enzymes and microorganisms. The characteristics of the support materials that ensure the viability of the immobilization are compared. In addition, the challenges and opportunities that arise from incorporating magnetic nanoparticles in immobilized systems are addressed.

Fabrication of substituted hydroxyapatite-starch-clay bio-composite coated titanium implant for new bone formation.

The clay/polymeric matrices have much attention from researchers in bio-medical applications due to their numerous uses. This study introduces new orthopedic titanium (Ti) implant with increasing bio-activity by treating the surface of the Ti implant with bio-compatible composite coating. Wollastonite (WST) clay combined minerals (Mg2+and Gd3+) substituted hydroxyapatite (HAP)/Starch composite was prepared using in-situ co-precipitation method. It was successfully coated on the orthopedic grade Ti plate by the Electrophoretic Deposition (EPD) method. The functionality, phase, morphology, and bio-activity analysis of the composite were evaluated by FT-IR, XRD, HR-TEM, and SEM analysis, respectively. The mechanical property, i.e., Vickers microhardness value of the MHAP/Starch/WST composite coated Ti plate, showed 242 ± 1.92 Hv. The in-vitro MG-63 osteoblast cells viability, differentiation, and Ca mineralization of MHAP/Starch/WST composite suggests that this new implant will be used for bone regeneration application after careful evaluation of in-vivo and clinical studies.

A Brief Review on Additive Manufacturing of Polymeric Composites and Nanocomposites.

In this research article, a mini-review study is performed on the additive manufacturing (AM) of the polymeric matrix composites (PMCs) and nanocomposites. In this regard, some methods for manufacturing and important and applied results are briefly introduced and presented. AM of polymeric matrix composites and nanocomposites has attracted great attention and is emerging as it can make extensively customized parts with appreciably modified and improved mechanical properties compared to the unreinforced polymer materials. However, some matters must be addressed containing reduced bonding of reinforcement and matrix, the slip between reinforcement and matrix, lower creep strength, void configurations, high-speed crack propagation, obstruction because of filler inclusion, enhanced curing time, simulation and modeling, and the cost of manufacturing. In this review, some selected and significant results regarding AM or three-dimensional (3D) printing of polymeric matrix composites and nanocomposites are summarized and discuss. In addition, this article discusses the difficulties in preparing composite feedstock filaments and printing issues with nanocomposites and short and continuous fiber composites. It is discussed how to print various thermoplastic composites ranging from amorphous to crystalline polymers. In addition, the analytical and numerical models used for simulating AM, including the Fused deposition modeling (FDM) printing process and estimating the mechanical properties of printed parts, are explained in detail. Particle, fiber, and nanomaterial-reinforced polymer composites are highlighted for their performance. Finally, key limitations are identified in order to stimulate further 3D printing research in the future.

Bacteriophage-Based Biosensing of Pseudomonas aeruginosa: An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains.

During the last decennium, it has become widely accepted that ubiquitous bacterial viruses, or bacteriophages, exert enormous influences on our planet's biosphere, killing between 4-50% of the daily produced bacteria and constituting the largest genetic diversity pool on our planet. Currently, bacterial infections linked to healthcare services are widespread, which, when associated with the increasing surge of antibiotic-resistant microorganisms, play a major role in patient morbidity and mortality. In this scenario, Pseudomonas aeruginosa alone is responsible for ca. 13-15% of all hospital-acquired infections. The pathogen P. aeruginosa is an opportunistic one, being endowed with metabolic versatility and high (both intrinsic and acquired) resistance to antibiotics. Bacteriophages (or phages) have been recognized as a tool with high potential for the detection of bacterial infections since these metabolically inert entities specifically attach to, and lyse, bacterial host cells, thus, allowing confirmation of the presence of viable cells. In the research effort described herein, three different phages with broad lytic spectrum capable of infecting P. aeruginosa were isolated from environmental sources. The isolated phages were elected on the basis of their ability to form clear and distinctive plaques, which is a hallmark characteristic of virulent phages. Next, their structural and functional stabilization was achieved via entrapment within the matrix of porous alginate, biopolymeric, and bio-reactive, chromogenic hydrogels aiming at their use as sensitive matrices producing both color changes and/or light emissions evolving from a reaction with (released) cytoplasmic moieties, as a bio-detection kit for P. aeruginosa cells. Full physicochemical and biological characterization of the isolated bacteriophages was the subject of a previous research paper.

Polysaccharides-Based Complex Particles' Protective Role on the Stability and Bioactivity of Immobilized Curcumin.

The curcumin degradation represents a significant limitation for its applications. The stability of free curcumin (FC) and immobilized curcumin in complex particles (ComPs) based on different polysaccharides was studied under the action of several factors. Ultraviolet-visible (UV-VIS) and Fourier-transform infrared (FTIR) spectroscopy proved the FC photodegradation and its role as a metal chelator: 82% of FC and between 26% and 39.79% of curcumin within the ComPs degraded after exposure for 28 days to natural light. The degradation half-life (t1/2) decreases for FC when the pH increases, from 6.8 h at pH = 3 to 2.1 h at pH = 9. For curcumin extracted from ComPs, t1/2 was constant (between 10 and 13 h) and depended on the sample's composition. The total phenol (TPC) and total flavonoids (TFC) content values increased by 16% and 13%, respectively, for FC exposed to ultraviolet light at λ = 365 nm (UVA), whereas no significant change was observed for immobilized curcumin. Antioxidant activity expressed by IC50 (µmoles/mL) for FC exposed to UVA decreased by 29%, but curcumin within ComPs was not affected by the UVA. The bovine serum albumin (BSA) adsorption efficiency on the ComPs surface depends on the pH value and the cross-linking degree. ComPs have a protective role for the immobilized curcumin.

Appreciating the First Line of the Human Innate Immune Defense: A Strategy to Model and Alleviate the Neutrophil Elastase-Mediated Attack toward Bioactivated Biomaterials.

Biointerface engineering is a wide-spread strategy to improve the healing process and subsequent tissue integration of biomaterials. Especially the integration of specific peptides is one promising strategy to promote the regenerative capacity of implants and 3D scaffolds. In vivo, these tailored interfaces are, however, first confronted with the innate immune response. Neutrophils are cells with pronounced proteolytic potential and the first recruited immune cells at the implant site; nonetheless, they have so far been underappreciated in the design of biomaterial interfaces. Herein, an in vitro approach is introduced to model and analyze the neutrophil interaction with bioactivated materials at the example of nano-bioinspired electrospun surfaces that reveals the vulnerability of a given biointerface design to the contact with neutrophils. A sacrificial, transient hydrogel coating that demonstrates optimal protection for peptide-modified surfaces and thus alleviates the immediate cleavage by neutrophil elastase is further introduced.

Biofilm Spreading by the Adhesin-Dependent Gliding Motility of Flavobacterium johnsoniae. 1. Internal Structure of the Biofilm.

The Gram-negative bacterium Flavobacterium johnsoniae employs gliding motility to move rapidly over solid surfaces. Gliding involves the movement of the adhesin SprB along the cell surface. F. johnsoniae spreads on nutrient-poor 1% agar-PY2, forming a thin film-like colony. We used electron microscopy and time-lapse fluorescence microscopy to investigate the structure of colonies formed by wild-type (WT) F. johnsoniae and by the sprB mutant (ΔsprB). In both cases, the bacteria were buried in the extracellular polymeric matrix (EPM) covering the top of the colony. In the spreading WT colonies, the EPM included a thick fiber framework and vesicles, revealing the formation of a biofilm, which is probably required for the spreading movement. Specific paths that were followed by bacterial clusters were observed at the leading edge of colonies, and abundant vesicle secretion and subsequent matrix formation were suggested. EPM-free channels were formed in upward biofilm protrusions, probably for cell migration. In the nonspreading ΔsprB colonies, cells were tightly packed in layers and the intercellular space was occupied by less matrix, indicating immature biofilm. This result suggests that SprB is not necessary for biofilm formation. We conclude that F. johnsoniae cells use gliding motility to spread and maturate biofilms.

The microalga Haematococcus lacustris (Chlorophyceae) forms natural biofilms in supralittoral White Sea coastal rock ponds.

MAIN CONCLUSION: Haematococcus lacustris inhabits supralittoral rock ponds and forms, under natural conditions, biofilms including layered cyanobacterial and fermentative microbial mats. Dry mats, formed under extremely stressful conditions, contained only haematocysts. Under favorable growth conditions, modeled for dry biofilms in vitro, microalgal free-living stages were detected. Haematococcus lacustris is the microalga known for its high potential to survive under a wide range of unfavorable conditions, particularly in the supralittoral temporal rock ponds of the White Sea. Previously, we described microbial communities containing H. lacustris in this region. In many cases, they were organized into systems exhibiting complex three-dimensional structure similar to that of natural biofilms. In this study, for the first time, we clarify structural description and provide microscopic evidence that these communities of H. lacustris and bacteria are assembled into the true biofilms. There are (1) simple single layer biofilms on the surface of rocks and macrophytic algae, (2) floccules (or flocs) not attached to a surface, (3) as well as stratified (layered) biofilms, wet, and dehydrated in nature. Being involved into primary organic production, H. lacustris and cyanobacteria are located exclusively in the upper layers of stratified biofilms, where they are capable to absorb sufficient for photosynthesis amount of light. The presence of acidic polysaccharides in the extracellular matrix revealed by specific staining with ruthenium red in the H. lacustris-containing microbial communities is a biochemical evidence of biofilm formation. Meanwhile, the presence of bacterial L-form is an ultrastructural confirmation of that fact. Under favorable conditions, modeled in vitro, H. lacustris from the dry microbial mats moves to the free-living states represented by vegetative palmelloid cells and motile zoospores. Owing to the fact that inside biofilms cells of microorganisms exist under stable conditions, we consider the biofilm formation as an additional mechanism that contributes to the survival of H. lacustris in the supralittoral zone of the White Sea.