Kinetic Study of Biopolymer (PHB) Synthesis in <Emphasis Type="Italic">Alcaligenes</Emphasis> sp. in Submerged Fermentation Process Using TEM

Poly-β-hydroxybuyrate (PHB) is a carbon—energy storage material which is accumulated as intracellular granule in variety of microorganism under nutrient starved conditions. Solid PHB is a biodegradable thermoplastic polymer and is utilizable in various ways similar to many conventional plastics. Ralstonia eutropha (Alcaligenes sp.), a gram negative bacteria accumulates PHB as insoluble granules inside the cells when nutrients other than carbon are limited. In this report effort has been made to analyze PHB granule synthesis inside Alcaligenes sp. NCIM 5085 by transmission electron microscopy and qualitative estimation of PHB was carried out by fourier transform infrared spectroscopy which provide better precision compared to other conventional techniques previously applied for PHB determination. Maximum PHB concentration of 2.20 ± 0.40 g/L and cell biomass of 3.42 ± 0.20 g/L was obtained after 48.0 h of fermentation. Leudking-Piret equation deduced mixed growth associated product formation which varies from earlier reports.     

Cultivation of Biogranules in a Continuous Flow Reactor at Low Dissolved Oxygen

This study investigated sludge granulation inoculated with various mixtures of aerobic and anaerobic sludge at low dissolved oxygen (DO; 0.3–0.6 mg/l) or aerobic (>2.5 mg/l) conditions in four parallel flow reactor systems. Formation of high-density coupled granules was achieved in the reactor system inoculated with anaerobic and aerobic sludge seeds (1:1 mass ratio) at low DO concentrations, with a mean size of 2.5 mm after only 27 days of cultivation. The highest ratio of protein (PN) to polysaccharide (PS; 3.3) was observed for the coupled sludge compared to granules cultivated under aerobic conditions. The PN/PS ratio correlated well with high hydrophobicity, low sludge volumetric index, and compact granular structure. Activity tests of the specific anaerobic and aerobic biomass confirmed that anaerobes and aerobes coexisted in the same coupled granule. Based on the optical microscopic and SEM observations, the process of coupled granule formation was proposed.     

The variability in nutrient composition of Anaerobic Digestate granules produced from high shear granulation

This study investigates the production of organic fertilizer using Anaerobic Digestate (as a nutrient source) and limestone powder as the raw materials. A two-level factorial experimental design was used to determine the influence of process variables on the nutrient homogeneity within the granules. Increasing the liquid-to-solid ratio during granulation resulted in increased granule nutrient homogeneity. Increasing the processing time and the impeller speed were also found to increase the nutrient homogeneity. In terms of nutrients release into deionized water, the granules effectively released both potassium and phosphate into solution.     

Production of artificial aggregates by granulation and carbonation of recycled concrete fines

There is a growing need to find ways to reuse fine concrete waste from the construction industry. In this study, recycled concrete fines were granulated and used as lightweight aggregates. Ladle slag, a steel industry residue, was used as a co-binder in different ratios (0, 10, 20, and 30%). The materials were blended and granulated, and then the granules were cured in three conditions: ambient condition, humidity chamber, and carbonation chamber. The results showed that the ladle slag content of 30% cured in a humidity chamber produced the strongest granules, with a crushing strength of 127 N, which was 135% greater than a commercial lightweight aggregate. The granules generally had satisfactory density and water absorption with a higher ladle slag content. Carbonation increased the granule strength with a low ladle slag content and decreased the granules’ water absorption. The improved physical and mechanical properties of carbonated granules are attributed to the formation of calcium carbonate during the carbonation process. The granules produced in this study show good potential for use as lightweight aggregates in the construction industry.

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Leaching of wood ash products aimed for spreading in forest floors--influence of method and L/S ratio

Use of biofuels in the form of logging residues is increasing in the European countries. This intensive forestry, where entire trees are removed from the felling sites, may contribute to a negative nutrient balance in the forest soil. Recycling of ash from the combustion of clean wood fuel, sometimes in combination with limestone or additives/binders, back into the forest soil could maintain the soil nutrient reservoir intact. Before spreading ash, it is important to determine its contents and, particularly, its decomposition pattern using reliable laboratory leaching tests. In this study, mineralogy and the leaching of Na, Ca, K, Mg, Mn, Al, Cu, Fe, P, and Zn from wood ash pellets and granules, produced both from green liquor sludge and fly ash, are examined by XRD and by subjecting these substances to three different laboratory leaching tests: upflow percolation (CEN/TS 14405), batch leaching (SS-EN12457), and a new Swedish leaching test using a magnetic stirrer. Mineral phases such as quartz, ettringite, calcite, gehlenite, and aphtitalite were identified in the ash granules and in the ash/green liquor sludge granules, by means of XRD. Six additional minerals were detected in the granules of ash only, and another six in the ash/green liquor sludge granules. At L/S 2, the batch leaching test resulted in the highest amounts of elements leached and the upflow percolation test the lowest. At L/S 10, both the batch leaching test and the upflow percolation test resulted in high amounts of elements leached. The batch leaching test at L/S 10 complies quite well with the percolation test and could be suitable for ash/green liquor sludge granule evaluation in daily practice. The magnetic stirrer test seems to underestimate the release potential of elements from granules. The batch test is simple to perform, and has the ability to dissolve 70-80% of the elements with the highest mobility from the materials under study.     

Formation of polyester blends by a recombinant strain ofPseudomonas oleovorans: Different poly(3-hydroxyalkanoates) are stored in separate granules

WhenPseudomonas oleovorans (GPo1) is grown on sodium octanoate under ammonium limiting conditions, it is able to accumulate a copolyester consisting of medium chain length 3-hydroxyalkanoic acids (PHA_m). 3-Hydroxybutyrate is only incorporated in trace amounts. WhenP. oleovorans is equipped with the PHB biosynthetic genes ofAlcaligenes eutrophus (GPo1[pVK101::PP1]), it forms a polyester containing major amounts of 3-hydroxybutyrate. The resulting polymer however is a blend of PHA_m and PHB, rather than a copolymer of 3-hydroxybutyrate and medium chain length 3-hydroxyalkanoic acids [11]. To establish whether PHA_m and PHB molecules are stored in the same or separate granules by this recombinantP. oleovorans strain, we studied polymer forming cells by freeze-fracture electron microscopy. This approach is possible because previous freeze-fracture electron microscopy studies on PHA_m and PHB accumulating strains have shown that PHA_m and PHB granules can be distinguished from each other: PHA_m granules from mushroom-like structures, whereas PHB granules from needle structures during freeze-fracturing. In this paper we show that stationary phase cells of GPo1[pVK101::PP1] contained both mushroom and needle-like structures, indicating that PHA_m and PHB chains were stored in separate granules. To be able to determine whether the separation of PHA_m and PHB is complete, the respective granules were separated on sucrose gradients. A total cell extract of GPo1[pVK101::PP1] which was subjected to sucrose gradient centrifugation revealed two white bands of different densities: the upper band with a density of 1.05 g/mL consisted exclusively of PHA_m granules, while the lower band with a density of 1.19 g/mL consisted of PHB granules only. Thus, when bacteria synthesize both PHA_m and PHB, the resulting polymer chains are segregated completely and stored in separate granules.     

Extruded Cornstarch-Glycerol-Polyvinyl Alcohol Blends: Mechanical Properties, Morphology, and Biodegradability

Elongation properties of extruded cornstarch were improved by blending with glycerol. Further blending of starch-glycerol with polyvinyl alcohol (PVOH) resulted in significant improvements in both tensile strength (TS) and elongation at break. Samples of starch-glycerol without PVOH equilibrated at 50% relative humidity had a TS of 1.8 MPa and elongation of 113%, whereas those containing PVOH had a TS and elongation of 4 MPa and 150%, respectively. Dynamic mechanical analysis (DMA) of starch-glycerol-PVOH blends showed that decreases in glass transition temperatures (T _g values) were proportional to glycerol content. Scanning electron microscopy (SEM) of fractured surfaces revealed numerous cracks in starch-glycerol (80:20) samples. Cracks were absent in starch-glycerol (70:30) samples. In both blends, many starch granules were exposed at the surface. No exposed starch granules were visible in blends with added PVOH. Starch-glycerol samples incubated in compost lost up to 70% of their dry weight within 22 days. Addition of PVOH lowered both the rate and extent of biodegradation.     

Solid State Reactions of Potato Starch with Urea and Biuret

Reaction of granular potato starch with urea and biuret resulted in the formation of products, which were soluble neither in cold nor boiling water. The net reaction was a monosubstitution of the hydrogen atom in one hydroxyl group in each D-glucose unit of starch with the either CO–NH₂ or CO–NH–CO–NH₂ moiety, respectively. Properties of the products, particularly these with urea, depended on the mode of reaction. Reactions were carried out in the microwave oven as well as with convection heating. The products retained the granular form of starch but a vast majority of granules were damaged. -Amylolysis of those materials revealed that their susceptibility to the enzyme increasing in the order: starch-amylolysis with simultaneous insolubility in water make these products suitable as ruminant fodder and, eventually, biodegradable material.     

Enhancing Reductive Dechlorination With Nutrient Addition

Enhanced reductive bioremediation (ERB) of halogenated organics has become widely accepted and implemented to reduce risks to human health and the environment posed by these compounds. The family of chlorinated ethenes (e.g., trichloroethene) is among the most common contaminants treated using ERB techniques. The number of microbial species able to fully dechlorinate ethenes is small, and many times their population numbers are low. As a result, many ERB practitioners have turned to the injection of designer bacteria within treatment zones as a means to accelerate the process. Otherwise, many remediation projects encounter “cis‐stall,'' or the accumulation of the cis‐1,2‐dichloroethene and vinyl chloride intermediates. This article discusses three unique case studies where the addition of a balanced macro‐ and micro‐nutrient source substantially accelerated the chlorinated ethene ERB process, independent of the electron‐donor substrate applied. © 2013 Wiley Periodicals, Inc.     

The Effect of Starch Composition on Structure of Foams Prepared by Microwave Treatment

A microwave technique was used to prepare foams from different potato starches in granular form, with varying amounts of amylose content, and water. In addition to native potato starch (PN), high amylose potato starch (HAP) and potato amylopectin (PAP) were used, as well as mixtures thereof. In all cases the native crystallinity of starch granules was lost upon microwave treatment and an amorphous material was created. An increased concentration of starch in the initial water dispersion resulted in a less dense foam structure. The potato amylopectin formed open cell foams, whereas increased amylose content, as in native potato starch, yielded a more compact structure with irregular pore shapes. The high amylose potato starch yielded a structure with hardly any porosity. Foaming experiments were done to compare pre-gelatinized and granular starches dispersed in water. The pre-gelatinisation did not affect the pore formation process. These experiments indicated that the molecular architecture of starch polymers is more important for foam formation than starch polymer organization in the granules. Studies of temperature profile and dry matter content during microwave treatment showed that water evaporates more rapidly from a high amylose starch solution than native potato starch and potato amylopectin solutions. Rheological measurements showed that the amylose solution had much lower viscosity than starch and amylopectin. This confirms that polymer – water interaction, such as in amylopectin solution, favours stabilization of bubbles formed upon boiling and evaporation of water, which yields high porosity materials.     

Thermoplastic Starch (TPS)/Polylactic Acid (PLA) Blending Methodologies: A Review

Polylactic acid (PLA) and thermoplastic starch (TPS) are biodegradable polymers of biological origin, and the mixture of these polymers has been studied due to the desirable mechanical properties of PLA and the low processing cost of TPS. However, the TPS/PLA combination is thermodynamically immiscible due to the poor interfacial interaction between the hydrophilic starch granules and the hydrophobic PLA. To overcome these limitations, researchers studied the modification, processing, and properties of the mixtures as a strategy to increase the compatibility between phases. This review highlights recent developments, current results, and trends in the field of TPS/PLA-based compounds during the last two decades, with the main focus of improving the adhesion between the two components. The TPS/PLA blends were classified as plasticized, compatible, reinforced and with nanocomposites. This article presents, based on published research, TPS/PLA combinations, considering different methods with significant improvements in mechanical properties, with promising developments for applications in food packaging and biomedicine.

     

Technology review and evaluation of different chemical oxidation conditions on treatability of PFAS

Per‐ and polyfluoroalkyl substances (PFAS) are a class of stable compounds widely used in diverse applications. These emerging contaminants have unique properties due to carbon–fluorine (C–F) bonds, which are some of the strongest bonds in chemistry. High energy is required to break C–F bonds, which results in this class of compounds being recalcitrant to many degradation processes. Many technologies studied that have shown treatment effectiveness for PFAS cannot be implemented in situ. Chemical oxidation is a demonstrated remediation technology for in situ treatment of a wide range of organic environmental contaminants. An overview of relevant literature is presented, summarizing the use of single or combined reagent chemical oxidation processes that offer insight into oxidation–reduction chemistries potentially capable of PFAS degradation. Based on the observations and results of these studies, bench‐scale treatability tests were designed and performed to establish optimal conditions for the formation of specific free radical species, including superoxide and sulfate radicals, via various combinations of oxidants, catalysts, pH buffers, and heat to assess PFAS treatment by chemical oxidants. The study also suggests the possible abiotic transformations of some PFAS when chemical oxidation is or was used for treatment of primary organic contaminants (e.g., petroleum or chlorinated organic compounds) at a site. The bench‐scale tests utilized field‐collected samples from a firefighter training area. Much of the available data related to chemical oxidation of PFAS has only been reported for one or both of the two more commonly discussed PFAS (perfluorooctane sulfonic acid and/or perfluorooctanoic acid). In contrast, this treatability study evaluates oxidation of a diverse list of PFAS analytes. The results of this study and published literature conclude that heat‐activated persulfate is the oxidation method with the best degradation of PFAS. Limited reduction of reported PFAS concentrations in this study was observed in many oxidation reactors; however, unknown mass of PFAS (such as precursors of perfluoroalkyl acids) that cannot be identified in a field collected sample complicated quantification of how much oxidative destruction of PFAS actually occurred.     

Recent Developments in Cleanup Technologies

Although the reaction mechanics are somewhat mysterious, the use of iron for in situ groundwater treatment has recently gained considerable attention and respect in the remediation industry. The basic scientific principles of both applications of iron have been known for over a century; however, both were nearly unheard of as remediation technologies five years ago. Both technologies have a strong potential for widespread use. They are commercially available, have been proven in field studies, are less expensive than traditional pump and treat technologies, and, in many types of groundwater systems, may be able to meet difficult-to-achieve groundwater treatment standards. As these technologies continue to undergo development, there could be considerably more aggressive applications used to treat ground-water containing high concentrations of chlorinated organics and DNAPLs.     

Biodegradable Blends Based on Starch and Poly(Lactic Acid): Comparison of Different Strategies and Estimate of Compatibilization

Finding plastic substitutes based on sustainability, especially for short-term packaging and disposable applications has aroused scientific interest for many years. Starch may be a substitute for petroleum based plastics but it shows severe limitations due to its water sensitivity and rather low mechanical properties. To overcome these weaknesses and to maintain the material biodegradability, one option is to blend plasticized starch with another biodegradable polymer. To improve both the compatibility between the main phases and the performance of the final blend, different compatibilization strategies are reported in literature. However, the relative efficiency of each strategy is not widely reported. This paper presents three different strategies: in situ (i) formation of urethane linkages; (ii) coupling with peroxide between starch and PLA, and (iiii) the addition of PLA-grafted amylose (A-g-PLA) which has been elaborated ex situ and carefully analyzed before blending. This study compares the effect of each compatibilization strategy by investigating mechanical and thermal properties of each blend. Compatibilizing behavior of the A-g-PLA is demonstrated, with a significant increase (up to 60%) in tensile strength of starch/PLA blend with no decrease in elongation at failure.     

Field methods and example applications for the Min-Trap® mineral sampler

Important reactive minerals are commonly created during in situ groundwater remediation activities; for example, iron sulfides formed during enhanced reduction approaches can abiotically degrade many chlorinated solvents. However, cost-effective tools to evaluate these treatment processes in field applications are limited and the collection of samples to evaluate in situ mineral formation is costly due to drilling requirements. The new passive Min-Trap sampler is a simple and cost-effective tool that can directly measure the formation of reactive minerals in situ without the need for additional drilling or soil core collection. The methods presented here describe how Min-Traps deployed in conventional monitoring wells can measure reactive minerals and how these minerals can be identified through commercially available analytical methods. Several examples are presented that show how Min-Traps can be used to characterize the rate and spatial variability of reactive mineral precipitation and these data may support operation and optimization decisions.     

Nuclear magnetic resonance logging: Example applications of an emerging tool for environmental investigations

Nuclear magnetic resonance (NMR) geophysical tools have been widely used in the petroleum exploration industry since the 1960s and have improved significantly in the last two decades. These tools can provide estimates of bulk porosity and fluid content, quantification of bound versus mobile fluids, and estimates of hydraulic conductivity (K). Although the size and cost of oil‐field tools historically limited their use for near‐surface applications, smaller and more economical downhole NMR logging tools are now available for detecting and characterizing the formation water content and K to support environmental and groundwater resource investigations. These tools can be deployed using direct‐push drilling techniques or they can be lowered into existing open borings or wells with nonconductive polyvinyl chloride casings and screens. In many cases, using the tool in existing wells offers a safer and more cost‐effective alternative compared to drilling new boreholes. For environmental investigations, NMR can provide useful high‐resolution quantitative hydrostratigraphic information that can provide additional valuable data to further inform and refine the conceptual site model. This paper highlights several NMR field investigations that demonstrate the viability of this technology as a site characterization tool for near‐surface investigations. NMR measurements were compared to data from lithologic logs, cone penetrometer testing data, and prior field hydraulic tests. Use of NMR to detect vadose zone water, including previously unidentified perched groundwater zones, provided hydrostratigraphic details that could not be gleaned from historical well drilling logs and were used to evaluate drainable pore water versus pore water bound in small pores by capillary forces or electrochemically clay‐bond water. NMR also produced K estimates similar to those from conventional hydraulic tests, but the improved vertical resolution from NMR provided additional information regarding the vertical heterogeneity of the formation along the entire length of the well or borehole. Additionally, bench‐scale tests are presented that confirm the capability for NMR to reliably detect and quantify light nonaqueous phase liquid saturation (specifically diesel fuel and weathered gasoline) in situ. The field tests combined with bench‐scale testing results affirm the applicability and potential for NMR as a practical characterization tool that should increasingly be utilized in environmental investigations.     

Utilisation of Supernatants of Pure Cultures of Streptomyces thermocarboxydus NH50 to Reduce Chromium Toxicity and Mobility in Contaminated Soils

Chromium is a heavy metal used in various industrial sectors. Improper handling and storage of chromium-laden effluents or wastes can lead to the pollution of the environment. The most toxic form is the more mobile one: hexavalent chromium Cr(VI). The reduction of Cr(VI) results in the immobilisation of chromium into its less toxic trivalent form Cr(III). This phenomenon may prevent the contamination of groundwater when the soil in the vadose zone is contaminated. Many bacteria have been isolated from contaminated soils and described to reduce Cr(VI) into Cr(III). A new Cr(VI)-reducing strain, identified as a Streptomyces thermocarboxydus,has been isolated and studied in our laboratories for its ability to reduce Cr(VI). This aerobic bacterium, in contrast to other genera described which mediate reduction via enzymes, produces reducing agents into the culture supernatants. Cr(VI) reduction by these substances is accelerated by the presence of small concentration of cupric ions (Cu²⁺). The reducing agent(s) can be easily recovered from the bacterial cultures and used as cell-free solution to treat contaminated soils by an in situ or ex situ processes.     

Morphological characteristics of waste polyethylene/polypropylene plastics during pyrolysis and representative morphological signal characterizing pyrolysis stages

In this work, the morphological characteristics of waste polyethylene (PE)/polypropylene (PP) plastics during their pyrolysis process were investigated, and based on their basic image changing patterns representative morphological signals describing the pyrolysis stages were obtained. PE and PP granules and films were used as typical plastics for testing, and influence of impurities was also investigated. During pyrolysis experiments, photographs of the testing samples were taken sequentially with a high-speed infrared camera, and the quantitative parameters that describe the morphological characteristics of these photographs were explored using the “Image Pro Plus (v6.3)” digital image processing software. The experimental results showed that plastics pyrolysis involved four stages: melting, two stages of decomposition which are characterized with bubble formation caused by volatile evaporating, and ash deposition; and each stage was characterized with its own phase changing behaviors and morphological features. Two stages of decomposition are the key step of pyrolysis since they took up half or more of the reaction time; melting step consumed another half of reaction time in experiments when raw materials were heated up from ambient temperatures; and coke-like deposition appeared as a result of decomposition completion. Two morphological signals defined from digital image processing, namely, pixel area of the interested reaction region and bubble ratio (BR) caused by volatile evaporating were found to change regularly with pyrolysis stages. In particular, for all experimental scenarios with plastics films and granules, the BR curves always exhibited a slowly drop as melting started and then a sharp increase followed by a deep decrease corresponding to the first stage of intense decomposition, afterwards a second increase – drop section corresponding to the second stage of decomposition appeared. As ash deposition happened, the BR dropped to zero or very low values. When impurities were involved, the shape of BR curves showed that intense decomposition started earlier but morphological characteristics remained the same. In addition, compared to parameters such as pressure, the BR reflects reaction stages better and its change with pyrolysis process of PE/PP plastics with or without impurities was more intrinsically process correlated; therefore it can be adopted as a signal for pyrolysis process characterization, as well as offering guide to process improvement and reactor design.     

Preparation and Properties of Thermoplastic Starch/Montmorillonite Nanocomposite Using N-(2-Hydroxyethyl)formamide as a New Additive

N-(2-Hydroxyethyl)formamide (HF) was synthesized efficiently and used as a new additive to prepare thermoplastic starch/montmorillonite nanocomposite (TPS/MMT). Here, HF acted as both plasticizer for TPS and swelling agent for MMT. The hydrogen bond interaction among HF, starch and MMT was proven by Fourier-transform infrared (FT-IR) spectroscopy. By scanning electron microscope (SEM), starch granules were completely disrupted. Atomic force microscopy demonstrated that partially exfoliated TPS/MMT nanocomposite was formed. The crystallinity of corn starch, MMT, HF-plasticized TPS (HTPS) and TPS/MMT nanocomposite was characterized by X-ray diffraction (XRD). Thermal stability of HTPS and TPS/MMT was determined by thermogravimetric analysis (TGA). The water resistance of TPS/MMT nanocomposite increased compared with that of pure HTPS. Tensile strengths of TPS/MMT nanocomposites were higher than those of HTPS, but just the reverse for elongations at break.     

Low‐concentration hydrogen peroxide decontamination for Bacillus spore contamination in buildings

Remediation and recovery efforts after a release of Bacillus anthracis (anthrax) spores may be difficult and costly. In addition, response and recovery technologies may be focused on critical resources, leaving the small business or homeowner without remediation options. This study evaluates the efficacy of relatively low levels of hydrogen peroxide vapor (HPV) delivered from off‐the‐shelf equipment for the inactivation of Bacillus spores within an indoor environment. Decontamination evaluations were conducted in a house using both Bacillus atrophaeus var. globigii (Bg; as surrogates for B. anthracis) inoculated on the carpet and galvanized metal as coupons and Geobacillus stearothermophilus (Gs) as biological indicators on steel. The total decontamination time ranged from 4 to 7 days. Using the longer exposure times, low concentrations of HPV (average levels below 20 parts per million) effectively inactivated Bg and Gs spores on the materials tested. The HPV was generated with commercial humidifiers and household‐strength hydrogen peroxide solutions. The presence of home furnishings did not have a significant impact on HPV efficacy. This simple, inexpensive, and effective decontamination method could have significant utility for remediation following a B. anthracis spore release, such as following a terrorist attack.