Testing for terrestrial and freshwater microalgae productivity under elevated CO2 conditions and nutrient limitation.

BACKGROUND: Microalgae CO2 fixation results in the production of biomass rich in high-valuable products, such as fatty acids and carotenoids. Enhanced productivity of valuable compounds can be achieved through the microalgae's ability to capture CO2 efficiently from sources of high CO2 contents, but it depends on the species. Culture collections of microalgae offer a wide variety of defined strains. However, an inadequate understanding of which groups of microalgae and from which habitats they originate offer high productivity under increased CO2 concentrations hampers exploiting microalgae as a sustainable source in the bioeconomy. RESULTS: A large variety of 81 defined algal strains, including new green algal isolates from various terrestrial environments, were studied for their growth under atmospheres with CO2 levels of 5-25% in air. They were from a pool of 200 strains that had been pre-selected for phylogenetic diversity and high productivity under ambient CO2. Green algae from terrestrial environments exhibited enhanced growth up to 25% CO2. In contrast, in unicellular red algae and stramenopile algae, which originated through the endosymbiotic uptake of a red algal cell, growth at CO2 concentrations above 5% was suppressed. While terrestrial stramenopile algae generally tolerated such CO2 concentrations, their counterparts from marine phytoplankton did not. The tests of four new strains in liquid culture revealed enhanced biomass and chlorophyll production under elevated CO2 levels. The 15% CO2 aeration increased their total carotenoid and fatty acid contents, which were further stimulated when combined with the starvation of macronutrients, i.e., less with phosphate and more with nitrogen-depleted culture media. CONCLUSION: Green algae originating from terrestrial environments, Chlorophyceae and Trebouxiophyceae, exhibit enhanced productivity of carotenoids and fatty acids under elevated CO2 concentrations. This ability supports the economic and sustainable production of valuable compounds from these microalgae using inexpensive sources of high CO2 concentrations, such as industrial exhaust fumes.

Rapid detection of φX-174 virus based on synchronous fluorescence of tryptophan.

The development of rapid methods for the detection of virus particles based on their intrinsic fluorescence (the native auto-fluorescence that originates from the non-labeled analyte) is challenging. Pure viruses may be detected in filtered solutions, based on the strong fluorescence of the amino acid tryptophan (Trp) in their proteins. Nevertheless, Trp also exists in high quantities in the hosts and host cultivation media. In this work, we developed a new method for the detection of the naked φX-174 virus. We show that a separation of φX-174 from its Escherichia coli host (grown on the standard cultivation medium nutrient agar) by simple extraction and filtration is not sufficient for its detection based on the intrinsic fluorescence since ~ 70% of the Trp fluorescence is derived from impurities. We formulate a new cultivation medium with a very low Trp concentration. We apply synchronous fluorescence measurements to show that no Trp fluorescence is detected in the extract solution upon incubation of this medium substrate with ammonium acetate extraction buffer. Finally, we apply synchronous fluorescence to detect φX-174 based on the spectral fingerprint of its native Trp content. Such a method is more rapid than usual traditional separation and detection methods which can take several hours and does not require any addition of labeling agents such as fluorescent dyes or antibodies for the detection. As other virus species contain Trp as one of the amino acids presents in their proteins, this method has the potential to apply to the detection of other viral species.

From the Titanic and other shipwrecks to biofilm prevention: The interesting role of polyphenol-protein complexes in biofilm inhibition.

Bacteria attach themselves either reversibly or irreversibly onto practically any surface in aqueous and other environments in order to reproduce, while generating extracellular polymeric substances (EPS) as a supportive structure for biofilm formation. Surfaces with a potential to prevent cellular attachment and aggregation (biofilm) would be extremely useful in environmental, biotechnological, medical and industrial applications. The scientific community is currently focusing on the design of micro- and nano-scale textured surfaces with antibacterial and/or antifouling properties (e.g., filtration membranes). Several serum and tissue proteins promote bacterial adhesion (for example, albumin, fibronectin and fibrinogen), whereas polyphenols form complexes with proteins which change their structural, functional and nutritional properties. For example, tannic acid, a compound composed of polygalloyl glucoses or polygalloyl quinic acid esters and several galloyl moieties, inhibits the growth of many bacterial strains. The present review is based on different nautical archaeology research data, and asks a simple but as yet unanswered question: What is the chemistry that prevents leather biodegradation by environmental bacteria and/or formation of biofilms? Future research should answer these questions, which are highly important for biofilm prevention.

Facilitated enumeration of the silicate bacterium Paenibacillus mucilaginosus comb. nov. (formerly Bacillus mucilaginosus) via tetrazolium chloride incorporation into a double agar-based solid growth medium.

Accurate enumeration of Paenibacillus mucilaginosus (formerly Bacillus mucilaginosus) bacterium from environmental samples on solid medium is challenging owing to its extensive extracellular polysaccharides (EPS) excretion. In the present study, P. mucilaginosus enumeration has been facilitated by a simple modification: addition of triphenyl tetrazolium chloride (TTC) to growth medium and application of a second soft agar layer. Results show distinctively better and accurate colonies' count. This method can be applied to all bacterial species that produce excessive EPS that may interfere with direct count.

Encapsulated Pseudomonas putida for phenol biodegradation: Use of a structural membrane for construction of a well-organized confined particle.

Phenols are toxic byproducts from a wide range of industry sectors. If not treated, they form effluents that are very hazardous to the environment. This study presents the use of a Pseudomonas putida F1 culture encapsulated within a confined environment particle as an efficient technique for phenol biodegradation. The innovative encapsulation technique method, named the "Small Bioreactor Platform" (SBP) technology, enables the use of a microfiltration membrane constructed as a physical barrier for creating a confined environment for the encapsulated culture. The phenol biodegradation rate of the encapsulated culture was compared to its suspended state in order to evaluate the effectiveness of the encapsulation technique for phenol biodegradation. A maximal phenol biodegradation rate (q) of 2.12/d was exhibited by encapsulated P. putida at an initial phenol concentration of 100 mg/L. The biodegradation rate decreased significantly at lower and higher initial phenol concentrations of 50 and up to 3000 mg/L, reaching a rate of 0.1018/d. The results also indicate similar and up to double the degradation rate between the two bacterial states (encapsulated vs. suspended). High resolution scanning electron microscopy images of the SBP capsule's membrane morphology demonstrated a highly porous microfiltration membrane. These results, together with the long-term activity of the SBP capsules and verification that the culture remains pure after 60 days using 16S rRNA gene phylogenetic affiliation tests, provide evidence for a successful application of this new encapsulation technique for bioaugmentation of selected microbial cultures in water treatment processes.

Co-cultivation of microalgae and nitrifiers for higher biomass production and better carbon capture.

The aim of this work was to study co-cultivation of nitrifiers with microalgae as a non-intrusive technique for selective removal of oxygen generated by microalgae. Biomass concentration was, at least, 23% higher in mixed-cultures where nitrifiers kept the dissolved oxygen concentration below 9.0µLL(-1) than in control Chlorella vulgaris axenic-cultures where the concentration of dissolved oxygen was higher than 10.0µLL(-1). This approach to eliminating oxygen inhibition of microalgal growth could become the basis for the development of advanced microalgae reactors for removal of CO2 from the atmosphere, and concentrated CO2 streams. CO2 sequestration would become a chemically and geologically safer and environmentally more sound technology provided it uses microalgal, or other biomass, instead of CO2, for carbon storage.

Preparation of biodegradable xanthan-glycerol hydrogel, foam, film, aerogel and xerogel at room temperature.

Polymers, hence hydrogels, pollute waters and soils accelerating environmental degradation. Environmentally benign hydrogels were made in water from biodegradable xanthan (X) and glycerol (G) at 22.5±2.5°C. Molar ratio [G]/[X]<3.0 was used to maximize crosslinking by mono-glycerol instead by poly-glycerol. XG-hydrogels were transformed into: XG-foams, XG-films, and XG-aerogel. Anionic character of XG-materials changes with changing [G]/[X] ratio. XG-films made from XG-hydrogels absorb up to 40 times more water than XG-films made from XG-foams. The films made from XG-foams and HCl do not dissolve in water during 48h. Making XG-materials is a no-waste process which decreases pollution, eliminates waste disposal costs, and minimizes energy expenses. XG-materials are suitable for both industrial and environmental applications including slow release and concentration of cations. XG-materials, made of xanthan, microbial polysaccharide, could also be used in applications targeting populations that do not consume meat or meat based products.

A simple medium modification for isolation, growth and enumeration of Acidithiobacillus thiooxidans (syn. Thiobacillus thiooxidans) from water samples.

High concentrations of H(2)S in groundwater are commonly removed using Biological Trickling Filter (BTF) that contains high numbers of biofilm immobilized sulfur oxidizing bacteria (mainly Thiobacillus thiooxidans). BTF performance requires continuous monitoring of these bacteria at several sampling points. The Most Probable Number (MPN) technique is at the moment the method of choice to enumerate viable T. thiooxidan cells under the above conditions. However, this method is extremely time-consuming (7-10days) and not always suitable for environmental monitoring. In the present study, Thiobacillus agar recommended for isolation and cultivation of Thiobacillus species by Spread plate method was modified by addition of bromocresol green (BCG) in order obtain a clear-cut resolution of the growing colonies resulting in similar or higher numbers compared to other methods. Visual emergence of bacterial colonies on the 3rd and 4th days, from the initial plating, was associated with sulfuric acid production, resulting in an unambiguous color change from blue to yellow, around each colony. This study revealed that BCG modified Thiobacillus agar is substantially time saving and much easier to infer compared to MPN technique.

Improvement of water quality using constructed wetland systems.

Constructed wetlands are among the recently proven efficient technologies for wastewater treatment. Compared with conventional treatment systems, constructed wetlands are low in cost, easily operated and maintained, and have a strong potential for application in developing countries, particularly by small rural communities. Nevertheless, the use of constructed wetlands for the improvement of drinking water quality (such as the purification of river water for drinking purposes) is still uncommon. Treatment technologies that use natural processes and/or passive components continue to be of interest to many segments of society for a wide variety of applications. This article summarizes information on the current methods used for water treatment using constructed wetland systems and presents several case studies.

Efficiency of phenol biodegradation by planktonic Pseudomonas pseudoalcaligenes (a constructed wetland isolate) vs. root and gravel biofilm.

In the last two decades, constructed wetland systems gained increasing interest in wastewater treatment and as such have been intensively studied around the world. While most of the studies showed excellent removal of various pollutants, the exact contribution, in kinetic terms, of its particular components (such as: root, gravel and water) combined with bacteria is almost nonexistent. In the present study, a phenol degrader bacterium identified as Pseudomonas pseudoalcaligenes was isolated from a constructed wetland, and used in an experimental set-up containing: plants and gravel. Phenol removal rate by planktonic and biofilm bacteria (on sterile Zea mays roots and gravel surfaces) was studied. Specific phenol removal rates revealed significant advantage of planktonic cells (1.04 × 10(-9) mg phenol/CFU/h) compared to root and gravel biofilms: 4.59 × 10(-11)-2.04 × 10(-10) and 8.04 × 10(-11)-4.39 × 10(-10) (mg phenol/CFU/h), respectively. In batch cultures, phenol biodegradation kinetic parameters were determined by biomass growth rates and phenol removal as a function of time. Based on Haldane equation, kinetic constants such as µ(max) = 1.15/h, K(s) = 35.4 mg/L and K(i) = 198.6 mg/L fit well phenol removal by P. pseudoalcaligenes. Although P. pseudoalcaligenes planktonic cells showed the highest phenol removal rate, in constructed wetland systems and especially in those with sub-surface flow, it is expected that surface associated microorganisms (biofilms) will provide a much higher contribution in phenol and other organics removal, due to greater bacterial biomass. Factors affecting the performance of planktonic vs. biofilm bacteria in sub-surface flow constructed wetlands are further discussed.

Whole cell imprinting in sol-gel thin films for bacterial recognition in liquids: macromolecular fingerprinting.

Thin films of organically modified silica (ORMOSILS) produced by a sol-gel method were imprinted with whole cells of a variety of microorganisms in order to develop an easy and specific probe to concentrate and specifically identify these microorganisms in liquids (e.g., water). Microorganisms with various morphology and outer surface components were imprinted into thin sol-gel films. Adsorption of target microorganism onto imprinted films was facilitated by these macromolecular fingerprints as revealed by various microscopical examinations (SEM, AFM, HSEM and CLSM). The imprinted films showed high selectivity toward each of test microorganisms with high adsorption affinity making them excellent candidates for rapid detection of microorganisms from liquids.

Combined chemical-biological treatment for prevention/rehabilitation of clogged wells by an iron-oxidizing bacterium.

Groundwater wells containing large concentrations of ferrous iron face serious clogging problems as a result of biotic iron oxidation. Following a short time after their start off, wells get clogged, and their production efficiency drop significantly up to a total obstruction, making cleanup and rehabilitation an economic burden. The present study was undertaken to test an experimental combined treatment (chemical and biological) for future prevention or rehabilitation of clogged wells. Sphaerotilus natans (an iron-oxidizing bacterium) freshly isolated from a deep well was grown to form biofilms on two systems: coupons and sand buried miniature wedge wire screen baskets. A combined chemical-biological treatment, applied at laboratory scale by use of glycolic acid (2%) and isolated bacteriophages against Sphaerotilus natans (SN1 and ER1-a newly isolated phage) at low multiplicity of infection (MOI), showed inhibition of biofilm formation and inactivation of the contaminant bacteria. In addition to complete inactivation of S. natans planktonic bacteria by the respective phages, earlier biofilm treatment with reduced glycolic acid concentration revealed efficient exopolysaccharide (EPS) digestion allowing phages to be increasingly efficient against biofilm matrix bacteria. Utilization of this combined treatment revealed clean surfaces of a model stainless steel wedge wire screen baskets (commonly used in wells) for up to 60 days.

The quality of drinking water stored in canteens of field soldiers as a potential source of enteric diseases.

Israel Defense Forces (IDF) guidelines for drinking water require the use of water only from sources that have been inspected and authorized by a medical expert. This study aimed to compare canteen water quality of two military units (infantry and armoured corps), to search for sources of possible microbial contamination and to look for any impact on gastrointestinal symptoms. Statistical analysis revealed that canteens of armoured corp soldiers were significantly more contaminated compared to those of infantry soldiers. Outdoor taps and water in trailers were found to harbour significantly higher numbers of microbial indicators compared to showers/lavatory sources; however, the numbers were much lower compared to canteens. Canteen water retention for more than one day revealed significantly increased numbers of examined microbial parameters, possibly due to secondary contamination or regrowth. Gastrointestinal symptoms were not significantly different between the two units despite the significant canteen water quality difference. An odds ratio evaluation was conducted on 45 exposure-illness combinations based on gastrointestinal symptoms, exposure and soldiers affiliation. Out of these 45 combinations only 14 resulted in odds ratio > 1, where 3 had high values (7.44, 7.46 and 11.2) suggesting a possible connection between diarrhoea and/or vomiting versus coliphages and faecal coliforms.

Sol-gel-based poliovirus-1 detector.

Hybrid sol-gel films were used to grow Buffalo Green Monkey kidney cell tissues, which were used for poliovirus-1 detection. It is shown that the sol-gel approach allows cutting the standard EPA procedure from 48 to 24h of detection time; that better visualization of the plaques is obtained; that a variety of stains, including fluorescence, can be used; and that the shelf life of the resulting plaques system is well over a year.

Photocatalytic inactivation of Flavobacterium and E. coli in water by a continuous stirred tank reactor (CSTR) fed with suspended/immobilised TiO2 medium.

A photocatalytic continuous stirred tank reactor (CSTR) was built at laboratory scale to inactivate two environmental bacteria strains (Flavobacterium and E. coli) in tap water. Several parameters were found to impact reactor efficiency. Bacterial initial concentration is an important factor in inactivation rate. After 30 minutes of irradiation at 10(8)-10(9) CFU mL(-1) starting concentration, a >5 log reduction was achieved while at 10(4)-10(6) CFU mL(-1) only a 2 log reduction was observed. Water hardness and pH have an important influence on the photocatalytic inactivation process. Soft water, with low Ca(+2) and Mg(+2) at low pH approximately 5.3 resulted in increased inactivation of Flavobacterium reaching >6 orders of magnitude reduction. E. coli and Flavobacterium at pH 5 were inactivated by 3 logs more as compared to pH 7 under similar conditions. pH below TiO2 isoelectric point (approximately 5.6) supports better contact between bacteria and anatase particles resulting in superior inactivation. TiO2 powder suspension was compared with immobilised powder in sol-gel coated glass beads in order to exclude the need for particles separation from the treated water. TiO2 suspension was more effective by 3 orders of magnitude when compared to coated glass beads. An interesting observation was found between the two bacterial strains based on their hydrophobicity/hydrophilicity balance. The more hydrophobic Flavobacterium compared to E. coli was inactivated photocatalytically by >3 logs more then E. coli in the first 30 minutes of irradiation interval. The results indicate the importance of the parameters involved in the contact between TiO2 particles and microorganisms that govern the successful inactivation rate in CSTR.

Bacteriophage ecology in a small community sewer system related to their indicative role in sewage pollution of drinking water.

In view of various studies looking for the merit of coliphages as indicators of water pollution with viruses originating from faecal material, a small agricultural community (population of approximately 1500 inhabitants of all ages, 2-3 km from Haifa) was selected in order to understand these bacteriophage ecology (F-RNA and somatic coliphages) in its sewer and oxidation pond system. Along the sewer lines, it was possible to isolate constantly both bacteriophage types (F-RNA and somatic coliphages) at 10(2)-10(4) plaque-forming units (pfu) ml(-1). The average numbers of somatic and F-RNA phages isolated from oxidation pond were 10(3)-10(4) pfu ml(-1); however, somatic coliphages were undetectable for several months (April-August). Significant high correlation (0.944 < R(2) < 0.99) was found between increased anionic detergent concentrations and F-RNA coliphage numbers. Infants less than 1 year old excreted both phage types and few only F-RNA coliphages (at high numbers > 10(5) pfu g(-1)) for up to 1 year. The excretion of F-RNA coliphages was highly linked to Escherichia coli F(+) harborage in the intestinal track as found in their faecal content. Finally, three bacterial hosts E. coli F(+), F(-) and CN(13) tested for survivability in sewage filtrate revealed that E. coli F(+) had the highest survivability under these conditions. Presence of somatic and F male-specific phages in sewer lines of a small community are influenced by several factors such as: anionic detergents, nutrients, temperature, source (mainly infants), shedding and survival capability of the host strain. Better understanding of coliphages ecology in sewer systems can enhance our evaluation of these proposed indicator/index microorganisms used in tracking environmental pollution of water, soil and crop contamination with faecal material containing enteric viruses.

Monitoring of effluent DOM biodegradation using fluorescence, UV and DOC measurements.

The potential of effluent DOM to undergo microbial degradation was assessed in batch experiments. Effluent samples from Haifa wastewater treatment plant and Qishon reservoir (Greater Haifa wastewater reclamation complex, Israel) were incubated either with effluent or soil microorganisms for a period of 2-4 months and were characterized by dissolved organic carbon contents (DOC), UV(254) absorbance and by fluorescence excitation-emission matrices. Three main fluorescence peaks were identified that can be attributed to humic/fulvic components and "protein-like" structures. During biodegradation, specific fluorescences (F/DOC) of the three peaks were increased at various extents, suggesting selective degradation of non-fluorescing constituents. In some cases increase in the effluent fluorescence (F) was observed thus proposing (i) the formation of new fluorescing material associated with DOM biodegradation and/or (ii) degradation of certain organic components capable of quenching DOM fluorescence. Based on the ratio between fluorescence intensity and UV(254), different biodegradation dynamics for fluorescent DOM constituents as compared with other UV-absorbing molecules was delineated. Overall, about 50% of the total DOM was found to be readily degradable such that residual resistant DOC levels were between 8 and 10 mg l(-1). Enhanced levels of residual DOM in effluent-irrigated soils may contribute to the DOM pool capable of carrying pollutants to groundwater.

A new sono-electrochemical method for enhanced detoxification of hydrophilic chloroorganic pollutants in water.

A new method for detoxification of hydrophilic chloroorganic pollutants in effluent water was developed, using a combination of ultrasound waves, electrochemistry and Fenton's reagent. The advantages of the method are exemplified using two target compounds: the common herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) and its derivative 2,4-dichlorophenol (2,4-DCP). The high degradation power of this process is due to the large production of oxidizing hydroxyl radicals and high mass transfer due to sonication. Application of this sono-electrochemical Fenton process (SEF) treatment (at 20 kHz) with quite a small current density, accomplished almost 50% oxidation of 2,4-D solution (300 ppm, 1.2 mM) in just 60 s. Similar treatments ran for 600 s resulted in practically full degradation of the herbicide; sizable oxidation of 2,4-DCP also occurs. The main intermediate compounds produced in the SEF process were identified. Their kinetic profile was measured and a chemical reaction scheme was suggested. The efficiency of the SEF process is tentatively much higher than the reference degradation methods and the time required for full degradation is considerably shorter. The SEF process maintains high performance up to concentrations which are higher than reference methods. The optimum concentration of Fe2+ ions required for this process was found to be of about 2 mM, which is lower than that in reference techniques. These findings indicate that SEF process may be an effective method for detoxification of environmental water.