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Biodegradation of phenol: a comparative study with and without applying magnetic fields.

The objective of this work was to study the effect of magnetic fields on the rate of phenol biodegradation using immobilized activated sludge. A recirculation flow bioreactor employing immobilized bacterial beads was used with phenol as the substrate to study the biodegradation process. This study was conducted by applying separately the north pole and the south pole magnetic fields to the bioreactor. Rate of dissolved oxygen consumption, phenol concentration and extracellular protein concentration were the parameters monitored during the process. It was observed that by applying a magnetic south pole to the process, biodegradation in the form of biological oxidation was enhanced. A 30% increase in biodegradation rate was obtained by applying a magnetic south pole of strength of 0.45 Tesla to the bioreactor with immobilized microbial beads as compared to the control. Magnetic north pole irradiation inhibited this type of biooxidation. This process has potential for biological treatment of organic wastes.

Bacteria↗

Antibiotic release from an experimental biodegradable bone cement.

An experimental biodegradable bone cement [poly(propylene fumarate)-methylmethacrylate] (PPF-MMA) has been compared in vivo with polymethylmethacrylate (PMMA) as a carrier agent for local release of antibiotics. This approach is potentially applicable to the treatment of chronic osteomyelitis where the clinical goal is to achieve sustained high concentrations of antibiotics locally in the infected bone. In our experiments, gentamicin- and vancomycin-impregnated cylindrical PMMA and PPF-MMA cement specimens were implanted subcutaneously in rats, and blood and wound fluid samples were obtained over a 2-week period. Antibiotic levels were determined using immunoassays, and microbiologic activity was confirmed with agar diffusion techniques. The biodegradable PPF-MMA cement achieved and maintained considerably higher wound antibiotic levels than did PMMA cement. Vancomycin levels for the PPF-MMA cement were greater than 20 times those for the PMMA cement at all sampling times from 24 h to 14 days. For both cements, the serum antibiotic concentrations remained safely below maximum levels recommended for parenteral therapy. Mechanical testing of the PPF-MMA cement showed that admixture of 3% by weight of antibiotic did not adversely affect material properties. We conclude that this experimental biodegradable bone cement (PPF-MMA) can be used as a carrier to achieve high sustained local levels and low serum levels of antibiotics. Because it is biodegradable and thus does not require a secondary procedure for removal, it has special potential for use in treatment of chronic osteomyelitis.

Administration, Topical↗

The crucial role of calcium interacting with soil pH in enhanced biodegradation of metam-sodium.

Enhanced biodegradation of soil-applied pesticides has long been correlated with soil pH above ca 6.5-7.5, but the possibility of confounding or interdependence with calcium, given that soil calcium concentration increases exponentially as pH rises above that range, has not previously been studied. Enhanced biodegradation of the broad-spectrum biocide metam-sodium was readily induced de novo in a naturally acid sandy soil (pH 4.2 measured in 0.01 M CaCl2) by multiple treatments, but only when the pH and calcium concentration were raised simultaneously using calcium carbonate (lime). Enhanced biodegradation was not induced when soil pH alone was raised with magnesium carbonate, nor when calcium alone was raised using calcium chloride. In limed sand treated monthly for 12 months, the degradation rate increased to where dissipation was complete within 24 h of application after the fifth metam-sodium treatment at pH 7.8 and after the eighth metam-sodium treatment at pH 6.8. Pesticide concentration was reduced, but not eliminated, at pH 5.8 and was unchanged at pH 4.8. When metam-sodium was applied bi- and tri-monthly, the degradation rate also increased when soil pH was raised with calcium carbonate, but to a lesser extent than with monthly applications. In an acid loam soil amended to the same pH values with calcium carbonate and treated monthly, there was no correlation between soil pH or calcium concentration and degradation. The results reveal the crucial interdependence of pH and calcium concentration in enhancement of biodegradation of soil-applied pesticides, but confirm that the phenomenon ultimately depends on interaction with soil type and frequency of application factors, all of which probably together act to affect the abundance, composition and activity of the soil microbial biomass.

Biodegradation, Environmental↗

Acute toxicity, mutagenicity, and estrogenicity of biodegradation products of bisphenol-A.

Biodegradation of bisphenol-A (BPA), which is known as an estrogenic chemical, proceeds via complicated metabolic routes and leads to formation of several kinds of biodegradation products. Through the major route BPA can be completely mineralized; however, p-hydroxyacetophenone (p-HAP), p-hydroxybenzaldehyde (p-HBAL), and p-hydroxybenzoic acid (p-HBA) are transiently accumulated at relatively high concentrations. On the other hand, degradation of BPA through the minor route tends to cause the accumulation of 2,3-bis(4-hydroxyphenyl)-1,2-propanediol and p-hydroxyphenacyl alcohol as the dead-end products. To fully assess the impact of BPA discharge into the environment, the considerable BPA degradation products p-HAP, p-HBAL, and p-HBA and the mixture of the dead-end products were examined for their acute toxicity, mutagenicity, and estrogenicity using the Daphtoxkit (Creasel Ltd.), umu test system, and yeast two-hybrid system, respectively. BPA was moderately toxic to Daphnia magna (48-h EC(50) was 10 mg/L) and weakly estrogenic, with activity that was 5 orders of magnitude lower than that of 17beta-estradiol in the yeast screen, though no mutagenicity was observed. All the tested BPA biodegradation products showed very low acute toxicity compared with BPA, and none was mutagenic. A slight estrogenic activity was detected only for p-HAP among the tested degradation products. It was concluded that biodegradation can remarkably reduce the toxic effects of BPA.

Animals↗

Classification of biodegradability by growth kinetic parameters.

On the basis of classical Monod equations for microbial growth, kinetic parameter values are assessed to enable the characterization of biodegradability as a substance property. Although growth is a microbial property, the substance selects those species from a mixed inoculum that can profit at most under the prevailing conditions. Hence it is concluded that the combination of observed cell yield Y approximately 0.5 g solids/g COD, maximum growth rate mu max > 0.3/day, and half-saturation constant Ks < 0.6 mg COD/liter provides a suitable set of borderline criteria for classification of chemicals as readily biodegradable. Of these parameters mu max is suitable for differentiating chemicals, whereas Y seems more or less equal for all readily degradable substances and Ks is not easily measurable with existing standard test methods. In this approach the density of specific microbes in the inoculum or during the test is excluded from the substance characterization and thus adaptation phenomena have no influence. From correlation studies and model calculations it can be expected that if the above parameters are estimated on the basis of standard tests for ready biodegradability, these tests will not produce false positives. Provided that the above parameters can be estimated, any other method than just the standard methods can be used to classify a substance as readily biodegradable.

Biodegradation, Environmental↗

Xenobiotic biodegradation test using attached bacteria in synthetic seawater.

The aerobic biodegradability of aniline, used as reference chemical, has been performed in synthetic seawater with attached biomass in a continuously fed reactor (biofilm chemostat reactor, BCR). Marine bacteria inocula came from local marine fish aquarium filters to limit the geographic and seasonal variations in quality. A pretreatment of these inocula combining 5-microns filtration and centrifugation was used to concentrate bacteria and remove organic carbon contamination of the test. The performances of the BCR were tested in comparison with simple shake flask tests. Among the different variables tested, the ratio S0/X0 (initial concentration of xenobiotic to initial density of the inoculum), the presence of dissolved oxygen, and the hydraulic residence time appear to be the key parameters controlling the length of the biodegradation process. On the other hand, the addition of a cosubstrate (easily biodegradable compound) does not provide advantages. Thus, marine biofilm chemostat reactors with a high density of attached bacteria (around 10(7) cells cm-2) and fed with synthetic seawater plus nitrogen provide good tools for screening biodegradability of chemicals in the marine environment.

Aniline Compounds↗

Shake-flask test for determination of biodegradation rates of (14)C-labeled chemicals at low concentrations in surface water systems.

A simple shake-flask surface water biodegradability die away test with (14)C-labeled chemicals added to microgram per liter concentrations (usually 1-100 microg/L) is described and evaluated. The aim was to provide information on biodegradation behavior and kinetic rates at environmental (low) concentrations in surface water systems. The basic principle of measurement was to determine evolved CO(2) indirectly from measurements of total organic activity in subsamples after stripping off their content of CO(2). Used with surface water alone the test simulates a pelagic environment and amended with sediments (0.1-1 dry weight/L) the test is intended to simulate a water environment with suspended solids (e.g., resuspended sediments). A protocol of the test used with the (14)C technique or with specific chemical analysis was recently developed by the International Organization for Standardization. Practical experience with the method is presented for a set of reference substances. These substances could be ranked in five groups of decreasing biodegradability: aniline>p-nitrophenol, 2, 4-dichlorophenoxyacetic acid>4-chloroaniline>maleic hydrazide, pentachlorophenol>atrazine. It was found that degradation rates and lag periods varied considerably among sampling sites and sometimes also among samples from the same site. No significant correlation could be established between degradation rates and microbial biomass estimates. Even small portions of added sediments greatly enhanced biodegradation of the absorbable compound pentachlorophenol, probably by providing sites for microbial attachment. Repeated tests indicated consistent degradation behavior for the readily degradable substances, whereas degradation sometimes stopped or failed with the more recalcitrant substances. A preadaptation step involving regular reinoculation with freshly collected surface water could, however, overcome the problems of false-negative results.

2,4-Dichlorophenoxyacetic Acid↗

Biodegradability of metronidazole, olaquindox, and tylosin and formation of tylosin degradation products in aerobic soil--manure slurries.

The use of veterinary drugs (primarily antibiotics) in animal husbandry harbors the risk that these compounds end up in the farmland when manure is used as fertilizer. The biodegradability of three compounds, olaquindox (OLA), metronidazole (MET), and tylosin (TYL), was simulated in soil--manure slurries with 50 g of soil per liter. Supplemental batch sorption tests revealed that insignificant amounts of OLA and MET were located in the soil phase, whereas only 0.1 to 10% of the added amounts of TYL remained in the liquid phase. This may reduce the bioavailability and thus biodegradation rates of TYL. Unidentified metabolites of OLA and TYL and four known TYL metabolites were detected using HPLC. However, none of these substances were seen to persist in the biodegradation experiments, indicating that OLA and TYL most likely were mineralized in the experiments. Neither the use of sandy or clayey soil nor the use of 0, 1, or 10% (V/V) of manure added to these soils had a significant effect on the degradation rates. Degradation half-lives for the primary degradation were 3.3--8.1 days for TYL, 5.8--8.8 days for OLA, and 13.1--26.9 days for MET. Based on comparisons of results obtained with the benchmark chemical aniline and degradation half-lives of this compound in nature, it was assessed that results obtained with the current test method slightly overestimate real-world biodegradation rates.

Aerobiosis↗

Responses of embryonic and larval inland silversides, Menidia beryllina, to a water-soluble fraction formed during biodegradation of artificially weathered Alaska North Slope crude oil.

Weathered Alaska North Slope crude oil (ANS 521) was stirred for 2 and 14 days in 20 per thousand salinity sterile seawater or for 14 days in seawater with nutrients and a group of three (GO3) microorganisms from Prince William Sound, Alaska, that were capable of biodegrading hydrocarbons. A total of 0.65 and 0.69 mg/L water soluble fraction (WSF) of neutral fraction hydrocarbons was recovered from the 2- and 14-day stirred sterile systems, respectively. In comparison, a total of 7.5 mg/L WSF neutral fraction hydrocarbons was recovered from systems containing ANS 521 that were stirred and biodegraded by the GO3 microbes for 14 days. Toxicity/teratogenicity tests were conducted with neutral fraction hydrocarbons recovered from the sterile and biodegraded systems using embryonic inland silversides, Menidia beryllina. Hydrocarbons from the sterile systems did not cause statistically significant teratogenic responses at concentrations of 1%, 10%, and 100% (w/v) of recovered fractions (redissolved in 20 per thousand salinity sterile seawater). Counts of heart contraction rates were significantly lower (alpha</=0.05) at the 100% WSF concentration of hydrocarbons on days 5 and 6 of embryogenesis. In contrast, recovered and redissolved neutral fraction hydrocarbons from ANS 521, that were biodegraded for 14 days, caused statistically significant (alpha</=0.05) teratogenic responses at the 1, 10, and 100% WSF concentrations. Measurement of heart contraction rates showed statistically significant (alpha</=0.05) reductions at the 100% WSF concentration on days 2 through 6 of embryogenesis, compared to controls.

Analysis of Variance↗

Effects of oxygen, nitrogen, and temperature on gasoline biodegradation in soil.

Biodegradation was considered to be a feasible approach to remediate petroleum hydrocarbon-contaminated soil from a site at the University of Idaho. Before a full-scale treatment process was designed, the biodegradative capacity of the soil's indigenous microorganisms was tested. Gas chromatography was used to measure gasoline vapor components in the headspace above the contaminated soils held in closed containers. In a study of biodegradation kinetics, gasoline degradation rates under various conditions (different soil cores, temperatures, oxygen concentrations, and nutrient concentrations) were tested. It was found that gasoline hydrocarbons could be biodegraded at relatively high rates after appropriate nutrient additions. An unexpected observation was that the optimal concentration of oxygen for the gasoline-degrading microorganisms in these soils was only 10%.

Biodegradation, Environmental↗

Posterolateral lumbar fusion using facet joint fixation with biodegradable rods: a pilot study.

Roentgen stereophotogrammetric analysis (RSA) was used to assess whether there is a potential for biodegradable rods crossing the denuded facet joints to increase the stability and healing rate of lumbar posterolateral fusions. Eleven consecutive patients with lumbosacral disc/facet joint degeneration had a posterolateral fusion augmented with 2- or 3.2-mm biodegradable rods passing perpendicularly through the center of the denuded facet joints. The patients were followed-up with RSA in supine and erect positions monthly from the 2nd to the 6th postoperative month, and again 1 year postoperatively. All seven L5-S1 fusions healed. Four cases were stable as defined by RSA within 3 months, two within 6 months, and one within 1 year. One L4-S1 fusion could not be evaluated by RSA. None of the remaining three L4-S1 fusions fully healed. In all three cases 1- to 3-mm intervertebral translations remained at 1 year. None of the 11 fusions showed any radiographic signs of osteolysis around the biodegradable rods. The promising results of this pilot study indicate that posterolateral L5-S1 fusion augmented with transarticular biodegradable rods crossing the denuded facet joints may yield rapid intervertebral stabilization and a high healing rate without any adverse rod effects. This may be due to enhanced initial fusion stabilization and/or increased ossification induced by the rods.

Adult↗

Primary biodegradation of a series of alkyl sulfosuccinates by mixed bacterial culture.

A mixed bacterial culture capable of primary biodegradation of sodium alkyl sulfosuccinates R1-OOC-CH(SO3Na)-CH2-COO-R2 was obtained from soil microorganisms by enrichment cultivation and adaptation in the presence of mono-n-dodecyl sulfosuccinate. Gram-negative psychrophilic bacteria with proteolytic, lipolytic and ammonifying activities were prevalent in the culture. The process of primary biodegradation of alkyl sulfosuccinates can be described by first-order reaction kinetics. The rate constants for linear esters were ascending in the order C4 < C5 < C6 (45 mumol/min per g cell protein) and further descending with increasing length of the carbon chain C6 > C8 >> C13. Substitution of cyclohexyl for n-hexyl group resulted in fourfold decrease in biodegradation rate. Terminal branching of alkyl chain does not affect the rate of primary biodegradation.

Adaptation, Biological↗

Respirometric analysis of the biodegradation of organic contaminants in soil and water.

Client-funded bench-scale investigations concerning the likelihood of successfully applying biological remediation to hazardous wastes must be cost-effective, and they usually need only determine if biodegradation is likely to occur on site. To assess the potential for stimulating biodegradation, biochemical oxygen demand (BOD) was used to continuously monitor bacterial respiration during growth on mixed organic wastes from contaminated water and soil. Continuously collected oxygen-consumption data provided information on the overall metabolic activity of the resident bacterial population and permitted direct observation of the cessation of microbial respiratory activity and, thus, the termination of aerobic degradation. The correlation of biological oxygen utilization with biodegradation was confirmed using independent analytical methods. Continuous, long-term BOD analysis was applied to bench-scale studies to assess the biodegradation of mixed organic wastes from contaminated sites and industrial waste effluents. This information was used to make an initial determination regarding the need to further explore bioremediation as a potential remedial-action technology using on-site, pilot-scale testing.

Bacteria↗

The binary treatment of aqueous metribuzin using anodic fenton treatment and biodegradation.

An advanced oxidation process, anodic Fenton treatment (AFT), and a mixed microbial culture were used to degrade metribuzin [4-amino-6-tert-butyl-3-methylthio-1,2,4- triazin-5(4H)-one], a broad-use triazinone herbicide. Complete and rapid removal of metribuzin was demonstrated. The appearance and subsequent degradation of metribuzin oxidation products--deaminated metribuzin (DA), diketo metribuzin (DK), as well as the production of deaminated diketo metribuzin (DADK)--were observed. To support the use of AFT as a chemical pretreatment, the ratio of 5-day biochemical oxygen demand (BOD5) as measured in the standard test to chemical oxygen demand (COD) was investigated, and an increase from 0.03, a nonbiodegradable solution, to 0.35, a biodegradable solution, was observed. This increase in biodegradability was associated with decreased metribuzin, DA, and DK concentrations and increased DADK concentration. AFT effluent was inoculated with either an enriched microbial culture or Polyseed, a commercially available inoculum. Although there was minimal biodegradation of the remaining metribuzin, there was a significant decrease in DA concentration in inoculated incubations compared with sterile controls after 5- and 10-minute AFT treatment. The enrichment inoculate appeared more adapted toward the less-oxidized, 5-minute-treated effluent, whereas the Polyseed culture, developed to degrade complex waste solutions, appeared to be more effective in a moreoxidized, 10-minute-treated, and potentially more complex effluent. This research supports the continued investigation of AFT and biodegradation as a binary treatment of aqueous pesticide wastes.

Agriculture↗

Estimation of contaminant depletion in unsaturated soils using a reduced-order biodegradation model and carbon dioxide measurement.

The objective of this study was to develop a reduced-order model of biodegradation in unsaturated soils that allows the estimation of contaminant depletion, using available on-line measurements. Hexadecane was chosen as a model compound for petroleum hydrocarbons. A two-compartment model was developed, decoupling the intrinsic biodegradation kinetics from limiting factors imposed by field conditions, such as oxygen transfer and contaminant bioavailability. Two new experimental protocols (one for the liquid phase and the other for the solid phase) were developed to monitor hexadecane depletion, hexadecane mineralization, total mineralization, and evolution of the degraders. Using the liquid-phase experiment, parameters of a Haldane kinetic model and yield coefficients were identified and used in the complete model of biodegradation in soil. Using the carbon dioxide production curve, a biocontact kinetic model was identified so that, despite the high sensitivity of the model outputs to variations in the parameters, hexadecane depletion could be correctly predicted with an average error on the entire time trajectory of about 8%. Moreover, the ratio between hexadecane mineralization and total mineralization remained constant after a brief transient period, indicating that hexadecane mineralization could be deduced from the total carbon dioxide measurement. Finally, the new model developed in this study allows real-time monitoring of contaminant biodegradation, using on-line carbon dioxide measurement.

Alkanes↗

Biodegradation of phenanthrene, spatial distribution of bacterial populations and dioxygenase expression in the mycorrhizosphere of Lolium perenne inoculated with Glomus mosseae.

Interactions between the plant and its microbial communities in the rhizosphere control microbial polycyclic aromatic hydrocarbons (PAH) biodegradation processes. Arbuscular mycorrhizal (AM) fungi can influence plant survival and PAH degradation in polluted soil. This work was aimed at studying the contribution of the mycorrhizosphere to PAH biodegradation in the presence of ryegrass (Lolium perenne L., cv. Barclay) inoculated with Glomus mosseae (BEG 69) by taking into account the structure and activity of bacterial communities, PAH degrading culturable bacteria as a function of the distance from roots. Ryegrass was grown in compartmentalized systems designed to harvest successive sections of rhizosphere in lateral compartments polluted or not with phenanthrene (PHE). Colonization of roots by G. mosseae (BEG 69) modified the structure and density of bacterial populations in the mycorrhizosphere, compared to the rhizosphere of non-mycorrhizal plants. G. mosseae increased the density of culturable heterotrophic and PAH degrading bacteria beyond the immediate rhizosphere in the presence of PHE, and increased the density of PAH degraders in the absence of the pollutant. Biodegradation was not significantly increased in the mycorrhizosphere, compared to control non-mycorrhizal plants, where PHE biodegradation already reached 92% after 6 weeks. However, dioxygenase transcriptional activity was found to be higher in the immediate mycorrhizosphere in the presence of G. mosseae (BEG 69).

Bacteria↗

Depolymerisation and biodegradation of a synthetic tanning agent by activated sludges, the bacteria Arthrobacter globiformis and Comamonas testosteroni, and the fungus Cunninghamella polymorpha.

Degradation of a synthetic tanning agent CNSF (a condensation product of 2-naphthalenesulfonic acid (2-NSA) and formaldehyde) by four activated sludges, two previously characterised bacterial strains, Arthrobacter sp. 2AC and Comamonas sp. 4BC, and the fungus Cunninghamella polymorpha, was studied in batch culture at 25 degrees C by determining the changes in the concentrations of CNSF and its component monomers and oligomers (n2-n11). The loss of individual oligomers was correlated with the length of the NSA-CH2 chain. Approximately 25% of the total CNSF was degraded (i.e. mineralised) by the microbes contained in the four activated sludges and by the two bacterial isolates but with different lag phases and at different overall rates. The decline in CNSF concentration was due almost entirely to the biodegradation of the monomers (34.3% of CNSF) and, in particular, 2-NSA (27% of CNSF). There was no change in the n2-n11 components. The growth of C. polymorpha, on the other hand, arose from extracellular depolymerisation of CNSF oligomers and the biodegradation of the lower molecular mass products. Between 38% and 42% of total CNSF was degraded by C. polymorpha at 25 degrees C. The order of oligomer degradation was inversely related to degree of polymerisation. Eighty percent and 90% of the n4 and n5 and 100% oligomers n6-n11 were degraded after 120 h. At a higher temperature (37 degrees C) oligomers n4-n11 were degraded completely after 120 h. A combination of biodegradation (75%) and sorption to fungal biomass (25%) accounted for the measured loss of all oligomers from the solution phase. The CNSF degradation rates and the volume of fungal biomass produced (and therefore the extent of biosorption) were dependent on the presence of a second carbon source (both optimum at glucose 5 g/l). This is the first report that identifies and distinguishes between depolymerisation, sorption and biodegradation processes in the removal of CNSF and its component oligomers. The use of combinations of the depolymerising fungus C. polymorpha, and the monomer-degrading bacteria, Arthrobacter sp. 2AC and Comamonas sp. 4BC, have potential for wastewater treatment.

Adsorption↗

Biodegradation of hexachlorocyclohexane (HCH) by microorganisms.

The organochlorine pesticide Lindane is the gamma-isomer of hexachlorocyclohexane (HCH). Technical grade Lindane contains a mixture of HCH isomers which include not only gamma-HCH, but also large amounts of predominantly alpha-, beta- and delta-HCH. The physical properties and persistence of each isomer differ because of the different chlorine atom orientations on each molecule (axial or equatorial). However, all four isomers are considered toxic and recalcitrant worldwide pollutants. Biodegradation of HCH has been studied in soil, slurry and culture media but very little information exists on in situ bioremediation of the different isomers including Lindane itself, at full scale. Several soil microorganisms capable of degrading, and utilizing HCH as a carbon source, have been reported. In selected bacterial strains, the genes encoding the enzymes involved in the initial degradation of Lindane have been cloned, sequenced, expressed and the gene products characterized. HCH is biodegradable under both oxic and anoxic conditions, although mineralization is generally observed only in oxic systems. As is found for most organic compounds, HCH degradation in soil occurs at moderate temperatures and at near neutral pH. HCH biodegradation in soil has been reported at both low and high (saturated) moisture contents. Soil texture and organic matter appear to influence degradation presumably by sorption mechanisms and impact on moisture retention, bacterial growth and pH. Most studies report on the biodegradation of relatively low (< 500 mg/kg) concentrations of HCH in soil. Information on the effects of inorganic nutrients, organic carbon sources or other soil amendments is scattered and inconclusive. More in-depth assessments of amendment effects and evaluation of bioremediation protocols, on a large scale, using soil with high HCH concentrations, are needed.

Aerobiosis↗