Study of microbial degradation of nonionic surface-active agents in designing technologies for purifying waste water

Studies of degradation of non-ionic surfactants (NISA) in a model purification plant of an original design demonstrated an high rate and depth of degradation processes compared with periodic cultivation of free or immobilized degrading strains. A virtually complete primary degradation (99-99.5%), with destruction of the oxyethyl moiety of the molecule, was observed. In addition, NISA molecules were degraded to a greater extent, including considerable degradation of the hydrocarbon radical, partial degradation of aromatic structures in Neonol, and utilization of biologically "unyielding" fractions of commercial NISA preparations: polyethylene glycol (PEG) and long-chain fractions of polymer homologues.     

Size Distribution and Amino Acid Chemistry of Base-extractable Proteins from Washington Coast Sediments

Proteinaceous components from four Washington coast margin sediments were extracted with base, fractionated into one of four size classes (<3 kDa, 3–10 kDa, 10–100 kDa, >100 kDa), and analyzed for their amino acid contents. Base-extracted material accounts for ~30% of the total hydrolyzable amino acids (THAA) and each size fraction has a unique composition, regardless of where the sediment was collected (shelf or upper slope). The <3 kDa size fraction (~10% of base-extractable THAA) is relatively enriched in glycine (~30 mol%), lysine (~5 mol%), and non-protein amino acids (~5 mol%). Glycine and non-protein amino acids are common degradation products, and lysine is very surface active. We suggest that the <3 kDa size fraction, therefore, represents a diagenetic mixture of fragments produced during the degradation of larger proteins. The 3–10 and 10–100 kDa size fractions (~10% and 42% of base-extractable THAA, respectively) have similar amino acid distributions dominated by aspartic acid (~30 mol%). Enrichments in Asp is likely due to both preservation of Asp-rich proteins and the production of Asp during degradation. The >100 kDa size fraction (~38% of base-extractable THAA) is not dominated by any particular amino acid and can not be modeled by mixing the amino acid compositions of the other size fractions. We propose that the larger size fractions (10–100 kDa and >100 kDa) represent intact, or near intact, proteins. Estimates of isoelectric points and relative hydrophobicity suggest the base-extractable proteins are primarily acidic and have globular structures. Statistical comparisons to several known proteins indicates that the base-extractable component is most similar to planktonic cytoplasmic proteins.     

Study of antioxidant activities of sulfated polysaccharides from Laminaria japonica

The composition, molecular weight and in vitro antioxidant activity of various sulfated polysaccharides obtained by anion exchange chromatography, acid hydrolysis and radical process degradation of the crude sulfated polysaccharide extracted from Laminaria japonica were compared. The low sulfated F-A2, with a peak-molecular weight (Mp) of 5–15 kDa, 14.5% sulfated ester and 21.8% glucuronic acid, exhibited a very strong antioxidant activity on superoxide and hydroxyl radicals, with activity even higher than that of large molecular weight fractions F-A and F-B. However, highly sulfated fractions with a peak-molecular weight below 15 kDa had much lower antioxidant activities than other fractions. These results indicated that the sulfate group of the low molecular weight fractions represents a physical block for the reaction with oxygen radicals. The chemical properties and antioxidant activities of sulfated polysaccharide fractions obtained by radical process degradation of crude sulfated polysaccharide were quite different from those obtained by acid hydrolysates. By radical process degradation, the high molecular weight was decreased to give LM2 (Mp 8 kDa) and LM1 (Mp 1.5 kDa), with a yield of 40% and 15%, respectively. LM2 was enriched with fucose and sulfated ester, while containing low amounts of glucuronic acid. The antioxidant activity showed that LM2 was unable to scavenge either superoxide or hydroxyl radical, which suggested that radical process degradation targeted mainly ascopyllan-like species rich in glucuronic acid, while the fraction rich in sulfated l-fucose remained unchanged. However, LM1 with Mp 1.5 kDa still retained apparent scavenging ability for superoxide radical, although it contained no glucuronic acid and certain amounts of galactose and mannose as main neutral sugars. These result suggest that the antioxidant activity of sulfated polysaccharides is apparently related not only to molecular weight and sulfated ester content, as previously determined, but also to glucuronic acid and fucose content.     

Urea degradation by picophytoplankton in the euphotic zone of Lake Biwa

The ability of photoautotrophic picoplankton Synechococcus to degrade urea was examined in the euphotic zone of Lake Biwa. Samples were divided into pico (0.2–2.0 μm) and larger (>2.0 μm) size fractions by filtration. The rates of urea degradation (the sum of the rates of incorporation of carbon into phytoplankton cells and of liberation of CO₂ into water) measured by radiocarbon urea were 8 and 17 μmol urea m⁻³ day⁻¹ in June and July, respectively, for the picophytoplankton in the surface water, and 196 and 96 μmol urea m⁻³ day⁻¹, respectively for the larger phytoplankton. The rates decreased with depth, somewhat similar to the vertical profiles of the photosynthetic rate. The urea degradation rates were obviously high under light conditions. In daylight, urea was degraded into two phases, carbon incorporation and CO₂ liberation, whereas in the dark it was degraded only into the CO₂ liberation phase. The contribution of picophytoplankton to total phytoplankton in urea degradation was high in the subsurface to lower euphotic layer. Urea degradation activity was higher in the picophytoplankton fraction than in the larger phytoplankton fraction. Shorter residence times of urea were obtained in the upper euphotic zone. The contribution of picophytoplankton to urea cycling was 4% to 35%. The present results suggest that the picophytoplankton Synechococcus is able to degrade urea and effectively makes use of regenerated urea as a nitrogen source in the euphotic layer, and that picophytoplankton play an important role in the biogeochemical nitrogen cycle in Lake Biwa. Received: June 25, 1998 / Accepted: February 10, 1999     

Green krill, the indicator of micro- and nano-size phytoplankton availability to krill

Size-fractionated chlorophyll-a concentrations of surface seawater were measured for pico-, nano-, and micro-size fractions (<2 μm, 2–10 μm, and >10 μm respectively) during commercial krill fishery operations in the waters north of the South Shetland Islands. The proportion of green krill (individuals discoloured due to active feeding on phytoplankton) had significant regressions with chlorophyll-a concentrations in micro- and nano-size fractions. Between these two fractions, chlorophyll-a concentration in the micro-size fraction showed the higher partial regression coefficient. This result shows the importance of phytoplankton larger than nano-phytoplankton, especially micro-phytoplankton, in terms of a phytoplanktonic food source for Antarctic krill in the natural environment. Accepted: 6 February 1999     

Microbial and cytoplasmic membrane-based potentiometric biosensors for direct determination of organophosphorus insecticides

Potentiometric biosensors for the determination of organophosphorus (OP) insecticides were developed by applying either immobilized whole cells or cytoplasmic membrane fractions of wild-type Flavobacterium sp. on the surface of a glass pH electrode. The ability of Flavobacterium sp. to degrade OP compounds as sole carbon source was demonstrated for parathion with a degradation rate of almost 100% after 30 min and for chlorpyrifos of 33% after 48 h incubation. The products of hydrolysis of these compounds, p-nitrophenol and 3,5,6-trichloro-2-pyridinol, were accumulated in the medium and not used as substrates for growth by Flavobacterium sp. In the course of hydrolysis, which is catalyzed by organophosphorus hydrolase, two protons are released for each substrate molecule hydrolyzed. This stoichiometry forms the electrochemical basis of the potentiometric biosensors. Direct determination without previous extraction of OP was carried out in a stirred measuring cell with a pH electrode as transducer. Poly(carbamoyl sulfonate) (PCS) prepolymer, a hydrogel with good adhesive properties, was used for immobilization of whole cells and membrane-associated organophosphorus hydrolase. The sensor with cytoplasmic membrane fractions was superior to the one with whole cells and showed a linear range for paraoxon from 0.01 to 0.47 mM and 3 weeks' working stability. Received: 11 February 2000 / Received revision: 25 May 2000 / Accepted: 26 May 2000     

Mechanical Properties of Mammalian and Fish Gelatins as a Function of the Contents of α-Chain, β-Chain,and Low and High Molecular Weight Fractions

Small-strain oscillatory measurements and size-exclusion chromatography coupled to multiangle laser light scattering were used to study the mechanical properties and the molecular weight distribution, respectively, of acid porcine skin gelatins (type A), lime bovine bone gelatins (type B), and cold water fish gelatins, while principal component analysis (PCA) and partial least squares regression were used to relate the mechanical properties with the molecular weight distribution. The present study suggests a linear relationship between the mechanical properties and the fractions of low molecular weight (LMW) molecules, α-chains, β-chains, and high molecular weight (HMW) molecules. The Bloom value for mammalian gelatin was positively correlated with the fractions of α-chains, β-chains, and HMW molecules and negatively correlated with the fraction of LMW molecules. The dynamic storage modulus for cold water fish gelatin was positively correlated with the fractions of β-chains and HMW molecules and negatively correlated with the fractions of LMW molecules and α-chains.     

Novel evidence of cytochrome P450-catalyzed oxidation of phenanthrene in <Emphasis Type="Italic">Phanerochaete chrysosporium</Emphasis> under ligninolytic conditions

The presence of cytochrome P450 and P450-mediated phenanthrene oxidation in the white rot fungus Phanerochaete chrysosporium under ligninolytic condition was first demonstrated in this study. The carbon monoxide difference spectra indicated induction of P450 (130 pmol mg⁻¹ in the microsomal fraction) by phenanthrene. The microsomal P450 degraded phenanthrene with a NADPH-dependent activity of 0.44 ± 0.02 min⁻¹. One of major detectable metabolites of phenanthrene in the ligninolytic cultures and microsomal fractions was identified as phenanthrene trans-9,10-dihydrodiol. Piperonyl butoxide, a P450 inhibitor which had no effect on manganese peroxidase activity, significantly inhibited phenanthrene degradation and the trans-9,10-dihydrodiol formation in both intact cultures and microsomal fractions. Furthermore, phenanthrene was also efficiently degraded by the extracellular fraction with high manganese peroxidase activity. These results indicate important roles of both manganese peroxidase and cytochrome P450 in phenanthrene metabolism by ligninolytic P. chrysosporium.     

Matrix effects in PIXE analysis of aerosols and ashes

A comparison of instrumental neutron activation analysis (INAA) and proton-induced X-ray emission (PIXE) results for sizefractionated atmospheric aerosols (“coarse” and “fine” fractions with an equivalent aerodynamic diameter of 2–10 Μm and < 2 Μm, respectively, or the PM10 fraction) showed that PIXE yielded significantly lower results for the PM10 and coarse fractions, especially for elements with a low Z resulting from a particle size effect. Somewhat lower PIXE results were also obtained for the fine fraction of atmospheric aerosols. A correction is also needed for irregularly shaped deposits of combustion aerosols collected by a cascade impactor in 11 size fractions ranging from 0.016 to 14.3 Μm, as well as for thick samples of fly and bottom ashes. An equivalent layer thickness (ELT) model is proposed to correct the matrix effects in PIXE. The approaches for the calculation of ELT using a comparison of PIXE and INAA results or by comparing PIXE results obtained using two different incident proton beam energies (1.31 and 2.35 MeV) are described. The correction for the ash pellets and irregular deposits are also discussed.     

Biodegradation of petroleum sludge and petroleum polluted soil by a bacterial consortium: a laboratory study

This article presents a study of the efficiency and degradation pattern of samples of petroleum sludge and polluted sandy soil from an oil refinery. A bacterial consortium, consisting of strains from the genera Pseudomonas, Achromobacter, Bacillus and Micromonospora, was isolated from a petroleum sludge sample and characterized. The addition of nitrogen and phosphorus nutrients and a chemical surfactant to both the samples and bioaugmentation to the soil sample were applied under laboratory conditions. The extent of biodegradation was monitored by the gravimetric method and analysis of the residual oil by gas chromatography. Over a 12-week experiment, the achieved degree of TPH (total petroleum hydrocarbon) degradation amounted to 82–88% in the petroleum sludge and 86–91% in the polluted soil. Gas chromatography–mass spectrometry was utilized to determine the biodegradability and degradation rates of n-alkanes, isoprenoids, steranes, diasteranes and terpanes. Complete degradation of the n-alkanes and isoprenoids fractions occurred in both the samples. In addition, the intensities of the peaks corresponding to tricyclic terpenes and homohopanes were decreased, while significant changes were also observed in the distribution of diasteranes and steranes.     

Microzooplankton and macrozooplankton glutamate dehydrogenase as indices of the relative contribution of these fractions to ammonium regeneration in the Gulf of Maine

Ammonium regeneration by micro- (35–153 µm) andmacrozooplankton (> 153 µm) was determined in the Gulfof Maine by measuring the activity of the excretory enzyme glutamatedehydrogenase (GDH) in various size fractions. GDH maxima weregenerally observed to correspond to the depth of the chlorophyllmaximum as previously reported in the Gulf of Mexico and inthe vicinity of the Nantucket Shoals. GDH activity of the microzooplanktonwas considerably lower than the macrazooplankton, suggestingthe microzooplankton made only a minor contribution (1–11%) to the total ammonium regenerated. These results were confirmedby biomass estimates made from counts of individual species.Ammonium excretion by both zooplankton fractions was estimatedto supply 5–31 % of the nitrogen requirements for primaryproduction, with an estimated 59–63% supplied by the verticaltransport of nitrate (new nitrogen) into the euphoric zone.     

Origin and cycling of riverine inorganic carbon in the Sava River watershed (Slovenia) inferred from major solutes and stable carbon isotopes

The Sava River and its tributaries in Slovenia represent waters strongly influenced by chemical weathering of limestone and dolomite. The carbon isotopic compositions of dissolved inorganic carbon (DIC) and suspended organic carbon (POC) fractions as well as major solute concentrations yielded insights into the origin and fluxes of carbon in the upper Sava River system. The major solute composition was dominated by carbonic acid dissolution of calcite and dolomite. Waters were generally supersaturated with respect to calcite, and dissolved CO₂ was about fivefold supersaturated relative to the atmosphere. The δ¹³C of DIC ranged from −13.5 to −3.3‰. Mass balances for riverine inorganic carbon suggest that carbonate dissolution contributes up to 26%, degradation of organic matter ∼17% and exchange with atmospheric CO₂ up to 5%. The concentration and stable isotope diffusion models indicated that atmospheric exchange of CO₂ predominates in streams draining impermeable shales and clays while in the carbonate-dominated watersheds dissolution of the Mesozoic carbonates predominates.     

Correlations of rates of insulin release from islets and plateau fractions for β-cells

Pancreatic β-cells in intact islets of Langerhans perfused with various glucose concentrations exhibit periodic bursting electrical activity (BEA) consisting of active and silent phases. The fraction of the time spent in the active phase is called the plateau fraction and appears to be strongly correlated with the rate of release of insulin from islets as glucose concentration is varied. Here this correlation is quantified and a theoretical development is presented in detail. Experimental rates of insulin release are correlated with “effective” plateau fractions over a range of glucose concentrations. There are a number of different models for BEA in pancreatic β-cells and a method is developed here to quantify the dependence of a glucose dependent parameter on glucose concentration. As an example, the plateau fractions computed from the Sherman-Rinzel-Keizer model are matched with experimental plateau fractions to obtain a relationship between the model's glucose-dependent parameter, β, and glucose concentration. Knowledge of the relationships between β and glucose concentration and between experimental measurements of rates of insulin release and plateau fractions permits the determination of theoretical rates of insulin release from the model.     

Size-fractionated chlorophyll a biomass and picophytoplankton cell density along a longitudinal axis of a temperate estuary (Southampton Water)

Size-fractionated chlorophyll a biomass and picophytoplanktoncell number distributions were investigated along a longitudinalaxis of Southampton Water estuary during autumn. Chlorophylla concentration in the >5µm and the 1–5 µmsize fractions was highest midway down the estuary, and decreasedboth in the landward and seaward directions. In contrast, chlorophylla biomass in the 0.2–1 µm size fraction showed nodecline towards the seaward end of the estuary. In agreementwith this observation, phycoerythrin-containing picocyanobacteriacell concentration showed a positive exponential-like relationshipwith salinity and eukaryotic picophytoplankton were also highestat high salinities. Expressed as a percentage of total, chlorophylla standing stock in both the 1–5 µ.m (4.4–28.7%)and the 0.2–1 µm size fractions (1.7–8.6%)was inversely correlated with total chlorophyll a concentration.Both these two fractions made a greater input to the total phaeopigmentconcentration than to the total pool of active chlorophyll a.     

A Comparison Between the ATPase and Proton Pumping Activities of Plasma Membranes Isolated from the Stele and Cortex of Zea mays Roots

Plasma membrane vesicles of high purity, determined by markerenzyme assays, were obtained by phase partitioning microsomalfractions from stelar and cortical tissues of Zea mays (cv.LG11) roots. ATP hydrolytic activities in both of the plasmamembrane fractions were inhibited by vanadate, SW26 and erythrosinB, but were insensitive to nitrate. Activity in both fractionsexhibited a marked pH optimum of 6·5 and displayed typicalMichaelis-Menten kinetics. A high substrate specificity wasapparent in both the stele and cortex plasma membrane fractions,while the lower fractions, after phase partitioning, showedlower specificity for nucleotide substrates. Specific activitiesof the stele (67·8 µmol Pi mg^–1 h^–1)and cortex (78·4 µmol Pi mg^–1 h–¹)plasma membrane H⁺ -ATPases were very similar. Proton pumping activities in microsomal membrane fractions fromstele and cortex were inhibited by nitrate and insensitive tovanadate. Homogenization of stele and cortex tissue in the presenceof 250 mol m^–3 KI resulted in microsomal fractions exhibitingvanadate-sensitive, nitrate-insensitive proton pumping activity,suggesting a plasma membrane origin for this activity. SW26was also an effective inhibitor of proton pumping activity,although results indicated an interaction between SW26 and thefluorescent probes quinacrine and acridine orange. The results are discussed in relation to models for the transportof ions into the stele and are consistent with a role for theH⁺ -ATPase activity in this process. Key words: ATPase, cortex, plasma membrane, stele, Zea mays     

Identification of Antihypertensive Peptides from Peptic Digest of Two Microalgae, Chlorella vulgaris and Spirulina platensis

The peptidic fractions that inhibited angiotensin I–converting enzyme (ACE) were separated from the peptic digests of 2 microalgae, Chlorella vulgaris and Spirulina platensis, by ion exchange chromatography and gel filtration. Oral administration of peptidic fractions into spontaneously hypertensive rats at 200 mg/kg of body weight resulted in marked antihypertensive effects. Further separation of the peptidic fractions by ODS high-performance liquid chromatography furnished the following active peptides: Ile-Val-Val-Glu (inhibitory against ACE with an IC₅₀ of 315.3 μM), Ala-Phe-Leu (63.8 μM), Phe-Ala-Leu (26.3 μM), Ala-Glu-Leu (57.1 μM), and Val-Val-Pro-Pro-Ala (79.5 μM) from C. vulgaris; Ile-Ala-Glu (34.7 μM), Phe-Ala-Leu, Ala-Glu-Leu, Ile-Ala-Pro-Gly (11.4 μM), and Val-Ala-Phe (35.8 μM) from S. platensis. Received July 7, 2000; accepted January 15, 2001     

Different forms of nitrogen in particle size fractions separated from two soils

Summary The possible use of particle size separation of soils in water was tested for preparing fractions rich in ‘unknown’-N, which constitutes between 41 and 46% of the total N. Ten particle size fractions ranging from <0.2 to>1,000.0 μm were separated from the A horizon of the Bainsville soil, an Orthic Humic Gleysol, and from the Bh horizon of the Armadale soil, a poorly drained Podzol. The distribution of N (hydrolyzable-, amino acid-, protein-, amino sugar-, ammonia-, ‘unknown’-and unidentified-N) in the initial soils and in each size fraction was determined. While particle size separation did not yield fractions which contained essentially only ‘unknown’-N and few known N-components, it was possible to isolate fractions which were either rich in amino acid-or protein-N or in NH₃-N. In general, the finer size fractions tended to be enriched in amino acid-N while NH₃-N was more prominent in the coarser fractions. The amino sugar-N content of all size fractions was low.     

Anaerobic degradation pathway and kinetics of domestic wastewater at low temperatures

The effect of temperatures below 20 °C (20, 15 and 10 °C) on the anaerobic degradation pathway and kinetics of domestic wastewater fractionated at different sizes was studied in a fluidized-bed batch reactor. The overall degradation pathway was characterized by a soluble fraction degrading according to zero-order kinetics and a colloidal fraction (between 0.45 and 4.5 μm) that first disintegrates into a particulate fraction smaller than 0.45 μm before finally degrading. The colloidal degradation processes follow a first-order kinetic. In contrast, suspended solids (bigger than 4.5 μm) degrade to soluble and colloidal fractions according to first-order kinetics. The colloidal fraction originating from suspended solids further degrades into soluble fraction. These soluble fractions have the same degradation kinetics as the original soluble fraction. The suspended solids degradation was highly affected by temperature, whereas the soluble fraction slightly affected and the colloidal fraction was not affected at all. On the other hand, the colloidal non-degradable fraction increased significantly with the decrease in temperature while the suspended solids slowly increased. The soluble non-degradable fraction was little affected by temperatures changes.     

Dialysability of Magnesium and Calcium from Hospital Duplicate Meals: Influence Exerted by Other Elements

Total and dialysable magnesium and calcium levels and corresponding dialysabilities were measured in duplicate meals (n = 108) during 36 consecutive days. The interaction exerted by other nutrients and energy on them was also performed. Total mean magnesium and calcium fractions of 113.9 ± 98.3 and 337.2 ± 278.9 mg/meal respectively, were found. The Mg and Ca levels supplied by meals are positively (p < 0.05) correlated with macronutrient contents (carbohydrates and proteins). The mean dialysable Mg and Ca fractions were 56.9 ± 36.3 and 127.4 ± 112.3 mg/meal (50.4 ± 13.2 and 37.8 ± 10.7% as dialysabilities, respectively). Total Mg and Ca levels are significantly correlated with corresponding element dialysabilities (p < 0.05). For both minerals, significant correlations between their total and dialysable fractions and between their dialysable level and dialysabilities were noted (p < 0.01). The mean Mg and Ca daily dietary intakes (DDI) were 341.7 ± 68.0 and 1,011.6 ± 424.4 mg/day, respectively. For Ca and Mg the existence of similarities in their behaviour in meals and absorptive processes has been found. Duplicate meals with raw vegetables are good sources of bioaccessible Mg. High Ca dialysability has been found in the analysed meals. The fish and products constitute a good source of bioaccessible Ca. Mg, Ca, zinc, and chromium levels enhanced significantly the Mg dialysability. The Ca dialysability rose significantly with dialysable Ca and chromium fractions (p < 0.05).     

The use of silica gel prepared by sol-gel method and polyurethane foam as microbial carriers in the continuous degradation of phenol

A mixed microbial culture was immobilized by entrapment into silica gel (SG) and entrapment/ adsorption on polyurethane foam (PU) and ceramic foam. The phenol degradation performance of the SG biocatalyst was studied in a packed-bed reactor (PBR), packed-bed reactor with ceramic foam (PBRC) and fluidized-bed reactor (FBR). In continuous experiments the maximum degradation rate of phenol (q _s ^max) decreased in the order: PBRC (598 mg l⁻¹h⁻¹) > PBR (PU, 471 mg l⁻¹h⁻¹) > PBR (SG, 394 mg l⁻¹h⁻¹) > FBR (PU, 161 mg l⁻¹h⁻¹) > FBR (SG, 91 mg l⁻¹ h⁻¹). The long-term use of the SG biocatalyst in continuous phenol degradation resulted in the formation of a 100–200 μm thick layer with a high cell density on the surface of the gel particles. The abrasion of the surface layer in the FBR contributed to the poor degradation performance of this reactor configuration. Coating the ceramic foam with a layer of cells immobilized in colloidal SiO₂ enhanced the phenol degradation efficiency during the first 3 days of the PBRC operation, in comparison with untreated ceramic packing. Received: 2 December 1999 / Revision received: 2 February 2000 / Accepted: 4 February 2000