Recycling of waste products in the induction of biofilms to remediate the visual impact generated by quartz mining

The present study was undertaken to investigate the viability of the use of two waste products, cheese whey and composted organic waste, as nutrient sources in the induction of biological films on quartz surfaces, with the final aim of reducing the visual impact generated by quartz mining. Experiments were carried in laboratory in which quartz samples were colonized with microorganisms (mainly cyanobacteria) forming biofilms. Previous studies have shown that a nutritional supplement must be added for good development of biofilms and, therefore, application of the two waste products was compared with application of the chemical nutrient medium on which these types of microorganisms are usually cultivated. Both products provided better results than the culture medium, in terms of the speed of formation of the biofilm, faster with the waste products, and the degree of cover of the brilliant white colour of the quartz, better masked by the biofilms formed when the waste products were applied as a darker biofilm was obtained.     

The quartz crystal microbalance as a novel means to study cell-substrate interactions In situ

The quartz crystal microbalance (QCM) was first introduced as a mass sensor in gas phase and in vacuum. Since oscillator circuits capable of exciting shear vibrations of quartz resonators under liquid loading have been developed, the QCM became accepted as a new, powerful technique to follow adsorption processes at solid-liquid interfaces in chemical and biological research. Lately, the QCM technique has attracted considerable interest as a novel means to monitor cell-substrate interactions of mammalian cells in vitro. Because the establishment and modulation of cell-substrate contacts is important for many physiological processes, and potent techniques to measure these interactions noninvasively are rare, the present review highlights applications of the QCM technique in this field. The suitability of the QCM device to monitor attachment and spreading of mammalian cells in real time has been well established. The QCM response is dependent on the individual cell type that is examined. In order to identify the sources for these cell-type-specific results of QCM readings, and to understand the information content of the signal, attempts have been made to decompose the overall QCM response into subcellular contributions. The aforementioned subjects, together with a condensed introduction into the QCM technology, are included in this article.     

Development of engineered biofilms on poly- L-lysine patterned surfaces

A technique has been developed to selectively attach bacteria to solid supports using poly-l-lysine. The patterned biofilms were labeled with green fluorescent protein (GFP) or a nucleic acid stain and imaged using both confocal microscopy and GFP stereomicroscopy. E. coli DH10B, E. coli MC1061, and Pseudomonas sp. GJ1 were selectively attached to regions coated with poly-l-lysine but not to uncoated regions. In contrast, E. coli DH5, W3110 and 33456 attached indiscriminately to the coated and uncoated regions of the surface. Those organisms that selectively attached to the poly-l-lysine coated regions formed biofilms twice as thick as the organisms that attached indiscriminately to the surface. This technique can be used for selectively patterning surfaces with genetically engineered microorganisms for biosynthesis of secondary metabolites and biodegradation or for developing a bacterial-based microscale medical diagnostic tool.     

Microwave capillary torch as a means for modifying the electrophysical characteristics of metal surfaces

An experiment layout based on a pulsed capillary microwave torch and making it possible to excite an explosive emission microplasma on a metal surface in open air is implemented for the first time. It is shown that a microrelief in the form of micron-size microcraters forms on the initially smooth surface under the action of microsparks. As a result, the maximum secondary electron emission yield σ_max decreases from ～2 for the untreated surface to ～0.4 for the rough treated surface and remains low for a long time when exposed to atmospheric air.     

Limited acid hydrolysis as a means of fragmenting proteins isolated upon ProteinChip array surfaces

ProteinChip array technology enables protein purification, protein profiling, and biomarker discovery on a convenient biochip platform. Traditional proteomic approaches towards protein identification rely upon the generation of peptides through the use of specific proteases. However, for a variety of reasons, the digestion of proteins bound to planar arrays by specific proteases, such as trypsin, has proven to be difficult, at times providing little or no protein digestion at all. Additionally, should more than one protein be present on the array surface, the digestion product consists of peptides from different proteins, adding another dimension of complexity to database mining approaches. These factors have driven our group to explore alternative means of on-chip protein digestion. In this article, we describe an approach to generate peptide maps by limited acid hydrolysis. Depending upon the adsorbed protein, this method requires between 500 femtomole to 5 picomole of protein for on-chip hydrolysis. Besides generating several internal peptide fragments, limited acid hydrolysis also has the advantage of generating peptide ladders from the N- or C-terminus of the protein. From these ladders, partial primary sequence of the protein can be directly derived when analyzed by a simple laser desorption/ionization mass spectrometer. Furthermore, tandem mass spectrometry can be performed on several internal peptide fragments, thus facilitating the identification of several proteins within a mixture. Based upon the preliminary results of this work, we continue to explore the possibility of using limited acid hydrolysis to identify unknown proteins captured on ProteinChip array surfaces.     

Laboratory experiments on the impact of biofilms on the plankton of a large river

1. Benthic organisms can have a strong effect on the plankton in rivers, although normally only members of the macrofauna are considered as important consumers. In the present study we conducted experiments on four different dates (in December, March, June and September) to assess the potential role of periphytic heterotrophic flagellates (HF), ciliates and rotifers in the control of potamoplankton (bacteria, algae, HF and ciliates). 2. Natural periphyton was established on the walls of circular flow channels by exposing them to river water (River Rhine, Germany). The experimental channels (with periphyton) and control channels (without periphyton) were filled with riverine water and the increase rates of planktonic bacteria, algae, HF and oligotrich ciliates were determined for the two treatments. 3. The abundance of periphytic ciliates and rotifers at the beginning of the four experiments showed large differences with low values in December and March, and high values in June and September. Dominant potential consumers of plankton were the heterotrich ciliate Stentor sp. and bdelloid rotifers. 4. The rates of increase of planktonic algae, HF and ciliates were significantly smaller in the presence of periphyton compared with those in their absence. Significant interactions between the treatment (with and without periphyton) and the time of experiment were found for the planktonic HF and algae, indicating that the impact of the periphyton varied temporarily. The planktonic groups responded differently to the periphyton with the planktonic HF showing the highest loss rate. Significant differences were also found among the loss rates of different HF groups and different diatom size classes. 5. These laboratory experiments demonstrate that periphytic ciliates and rotifers are potentially important consumers of different planktonic groups. The quantitative impact of periphyton on plankton with respect to the selective feeding needs further attention.     

Site-specific variation in Antarctic marine biofilms established on artificial surfaces

The community structure and composition of marine microbial biofilms established on glass surfaces was investigated across three differentially contaminated Antarctic sites within McMurdo Sound. Diverse microbial communities were revealed at all sites using fluorescence in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) techniques. Sequencing of excised DGGE bands demonstrated close affiliation with known psychrophiles or undescribed bacteria also recovered from the Antarctic environment. The majority of bacterial sequences were affiliated to the Gammaproteobacteria, Cytophaga/Flavobacteria of Bacteroidetes (CFB), Verrucomicrobia and Planctomycetales. Principal components analysis of quantitative FISH data revealed distinct differences in community composition between sites. Each of the sites were dominated by different bacterial groups: Alphaproteobacteria, Gammaproteobacteria and CFB at the least impacted site, Cape Armitage; green sulfur and sulfate reducing bacteria near the semi-impacted Scott Base and Planctomycetales and sulfate reducing bacteria near the highly impacted McMurdo Station. The highest abundance of archaea was detected near Scott Base (2.5% of total bacteria). Multivariate analyses (non-metric multidimensional scaling and analysis of similarities) of DGGE patterns revealed greater variability in community composition between sites than within sites. This is the first investigation of Antarctic biofilm structure and FISH results suggest that anthropogenic impacts may influence the complex composition of microbial communities.     

Bacterial diversity in biofilms formed on condenser tube surfaces in a nuclear power plant

To elucidate the bacterial diversity in biofilms formed on a condenser tube from a nuclear power plant, 16S rRNA gene sequences were examined using a PCR-cloning-sequencing approach. Twelve operational taxonomic units were retrieved in the clone library, and the estimated species richness was low (13.2). Most of the clones (94.7%) were affiliated with α-Proteobacteria; Planctomycetes and γ-Proteobacteria were much rarer. Interestingly, except for one clone belonging to Pseudoalteromonas, most of the sequences displayed sequence similarities <97% of those of the closest type strains. Based on 16S rRNA phylogenetic analysis, most bacteria were assigned to novel taxa above the species level. The low species richness and unusual bacterial composition may be attributable to selective pressure from chlorine in the cooling water. To prevent or control bacterial biofilms in cooling circuits, additional studies of the physiology and ecology of these species will be essential.     

Uncoated quartz resonator as a universal biosensor

We studied dynamic processes in drying drops of model protein-salt solutions, using an uncoated quartz resonator as a biosensor. To measure these processes we developed a method based on recording the dynamics of the Acoustic-Mechanical Impedance (AMI) of a drop as it dried on the surface of a quartz resonator oscillating at a resonant frequency of 60 kHz. The aim of this work was to highlight the role of some components of serum in self-organization processes. Human serum albumin (HSA), fibronectin (Fn), immunoglobulin G (IgG), immunoglobulin M (IgM), bovine serum albumin (BSA), sodium chloride (NaCl), Potassium Chloride (KCl), and nonionic surfactant O(CH(2)CH(2))(n)CH(2)CH(2)OH were used as components of the tested solutions. It was shown that dynamics of the AMI in drying drops were closely related to liquid composition. This approach allowed us to distinguish with good accuracy solutions in which one or more components (proteins or salts) were replaced by other components with the same mass concentration. We assumed that these differences were due to different surface properties and native functions of proteins, and different positions of salts in the Hofmeister line. Our preliminary work demonstrated that the dynamics of phase transitions in drying drops of serum could be used as an informative parameter for medical diagnostics. In this study, we highlight some positions in this cause-effect chain.     

Campylobacter jejuni as a secondary colonizer of poultry biofilms

Aims: The objective of this study was to determine if survival of culturable Campylobacter jejuni outside the host was increased by entrapment in pre-established biofilms. Methods and Results: Campylobacter jejuni was inoculated into four biofilm populations isolated from poultry environments and cultured at three temperatures. Survival of culturable Camp. jejuni in some pre-established biofilms was extended vs survival of culturable Camp. jejuni in broth. But some biofilms were detrimental to survival of culturable Camp. jejuni. Denaturing gradient gel electrophoresis analysis indicated differences in bacterial profiles depending on initial source and temperature of culturing, which may have had impacts on survival of culturable Camp. jejuni. Further investigation showed no evidence of interspecies cell signalling indicating that secondary colonization was only physical. Conclusions: The results of this study show Camp. jejuni’s attachment to surfaces is facilitated by pre-established biofilms and survival of culturable Camp. jejuni may be extended in some pre-established biofilms, but these biofilms do not fully explain long-term survival of culturable Camp. jejuni outside hosts. Significance and Impact of the Study: This study provides new information concerning survival of culturable Camp. jejuni outside the host and shows biofilms may be important in transmission and prevalence of Camp. jejuni.     

Bacterial diversity in biofilms on external surfaces of historic buildings in Porto Alegre

Summary Bacteria, along with other microorganisms, are present on and in rocks or historic stone monuments and can cause biodeterioration. Gram-positive bacteria, although present in lower numbers, are principally responsible for the damage caused. Four churches and the Vice-Governor’s office in Porto Alegre, all buildings of historic importance were studied, using traditional microbiological methods, with the aim of assessing the microdiversity on their external surfaces. A large number of microorganisms was found in each biofilm. Cell morphology varied at different points and with season. Most of the gram-positive bacteria were of the Bacillus genus, which are readily able to survive the dry conditions on these exposed surfaces. The isolates with the highest deteriorating ability, producing acids and surfactants with autoemulsifing power, were Bacillus isolates B4, B6 and B10 from Priest groups II, I and III, respectively. These were evaluated.     

Detachment ofPseudomonas fluorescens from biofilms on glass surfaces in response to nutrient stress

The effects of glucose and nitrogen depletion on the colonization of glass Petri plates byPseudomonas fluorescens were studied in batch culture. Colonization of the surfaces was initiated before colonization of the bulk phase, and biofilm formation was observed. This resulted in an apparent lag in the batch growth curve for the cell suspension. The lag phase was an artifact caused by the partitioning of cells between the bulk and solid phase of the culture and was not due to a reduction in the growth rate of unattached cells. The specific growth rate of the unattached cells (0.331 hour^–1) was almost twice that determined for the total population (0.171 hour^–1). Consequently the growth rate of biofilm-forming bacteria cannot be determined in batch culture unless the growth of both attached and unattached cells is monitored, and batch growth curves may contain artifacts due to the formation and dispersion of biofilms. The depletion of either glucose or nitrogen led to the active detachment of cells from the biofilm. An increase in the hydrophobicity of unattached cells was noted on depletion of carbon. This increase was the result of emigration of cells from the surface into the bulk phase.Paper contribution number 128, Centre de Rechcrches Alimentaires de St Hyacinthe.     

Biofouling on polymeric heat exchanger surfaces with E. coli and native biofilms

The biofouling affinity of different polymeric surfaces (polypropylene, polysulfone, polyethylene terephthalate, and polyether ether ketone) in comparison to stainless steel (SS) was studied for the model bacterium Escherichia coli K12 DSM 498 and native biofilms originating from Rhine water. The biofilm mass deposited on the polymer surfaces was minimized by several magnitudes compared to SS. The cell count and the accumulated biomass of E. coli on the polymer surfaces showed an opposing linear trend. The promising low biofilm formation on the polymers is attributed to the combination of inherent surface properties (roughness, surface energy and hydrophobicity) when compared to SS. The fouling characteristics of E. coli biofilms show good conformity with the more complex native biofilms investigated. The results can be utilized for the development of new polymer heat exchangers when using untreated river water as coolant or for other processes needing antifouling materials.     

Behavior,activities, and effects of bacteria on synthetic quartz monocrystal surfaces

Two strains ofBacillus sp. and a strain ofBrevibacterium sp., originally isolated from a natural quartzite surface, were characterized and employed as test strains with several methods: acridine orange fluorochromation and epifluorescence microscopy were used for detection of individual cells; scanning and transmission microscopy for studying attachment behavior; replica techniques in combination with electron microscopy for following surface interaction effects; and chemical analysis of SiO₂ for detecting possible silica leaching activities. The experimental results clearly showed that the three test strains were able to attach to and grow on the precleaned quartz surfaces. Attachment modes were either by direct sorption mechanisms (Brevibacterium sp. S) or the production of adhesive polymers (Bacillus sp. U andBacillus sp. W). In short-term contact incubation experiments with rich media, neither quartz crystal surface structures nor bacterial cell surfaces appeared to be changed. Likewise, significant biochemical dissolution and mechanical dislocation of SiO₂ (which would have indicated rapid bacterial weathering activities) could not be detected. The importance of quartz purity and crystalline structure for the initiation of weathering processes is discussed.     

Gas-phase removal of biofilms from various surfaces using carbon dioxide aerosols

The present study evaluated the removal of Escherichia coli XL1-blue biofilms using periodic jets of carbon dioxide aerosols (a mixture of solid and gaseous CO₂) with nitrogen gas. The aerosols were generated by the adiabatic expansion of high-pressure CO₂ gas through a nozzle and used to remove air-dried biofilms. The areas of the biofilms were measured from scanning electron micrographs before and after applying the aerosols. The removal efficiency of the aerosol treatment was measured with various air-drying times of the biofilms before the treatment, surface materials, and durations of CO₂ aerosols in each 8-s aerosol–nitrogen cleaning cycle. Nearly 100% of the fresh biofilms were removed from the various surfaces very reliably within 90 s. This technique can be useful for removing unsaturated biofilms on solid surfaces and has potential applications for cleaning bio-contaminated surfaces.     

Ciliates as engineers of phototrophic biofilms

1. Phototrophic biofilms consist of a matrix of phototrophs, non‐photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the ‘engineering’ effects of these ciliates on the spatial heterogeneity of phototrophic biofilms are poorly studied. 2. We studied the potential engineering effects of two ciliates, Urostyla sp. and Paramecium bursaria, on the spatial heterogeneity of synthetic multispecies biofilms. Biomass of phototrophic organisms, EPS and bacteria was analysed three dimensionally using confocal laser scanning microscopy. Spatial heterogeneity and cover of the phototrophs, bacteria and EPS were determined at several depths within the biofilm. 3. Ciliate species did not interfere with the overall development of phototrophic microorganisms, because the thickness of the biofilm was equal whether the ciliates were present or not, even though their abundance did affect spatial heterogeneity of biofilm components. When Urostyla was present, it reduced aggregation in EPS and bacteria and increased EPS biovolume. This implies a local facilitating effect of ciliates on photosynthetic activity. Biofilms to which Paramecium was added did not differ from controls in terms of phototrophs, EPS cover and biovolume. Nevertheless, ciliates affected the spatial heterogeneity of these components as phototrophs and EPS became more evenly distributed. 4. This study shows that ecosystem engineering by organisms does not only occur at large spatial scales, as in grasslands and estuaries, but also plays a role at the microscopic scale of biofilms. This effect on spatial heterogeneity was not driven by substantial exploitation of biofilm components, but via the subtle engineering effects of ciliates.     

Gas-phase removal of biofilms from various surfaces using carbon dioxide aerosols

The present study evaluated the removal of Escherichia coli XL1-blue biofilms using periodic jets of carbon dioxide aerosols (a mixture of solid and gaseous CO(2)) with nitrogen gas. The aerosols were generated by the adiabatic expansion of high-pressure CO(2) gas through a nozzle and used to remove air-dried biofilms. The areas of the biofilms were measured from scanning electron micrographs before and after applying the aerosols. The removal efficiency of the aerosol treatment was measured with various air-drying times of the biofilms before the treatment, surface materials, and durations of CO(2) aerosols in each 8-s aerosol-nitrogen cleaning cycle. Nearly 100% of the fresh biofilms were removed from the various surfaces very reliably within 90?s. This technique can be useful for removing unsaturated biofilms on solid surfaces and has potential applications for cleaning bio-contaminated surfaces.