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Comparative biomonitoring study of workers at a waste disposal site using cytogenetic tests and the comet (single-cell gel) assay.

Workers exposed to environmental pollutants at a waste disposal site were studied for genotoxic effects with cytogenetic tests and the comet (alkaline single-cell gel) assay. Analyses were performed on peripheral blood samples of 44 workers at a waste disposal site (DM) and 47 subjects of a control group (VE) matched for gender, age, and smoking habits. Chromosomal aberrations were evaluated in 1,000 lymphocytes per individual, sister chromatid exchanges in 50 cells, and DNA migration (tail moment) was determined in 100 leukocytes. Structural chromosome aberrations were more frequent in DM than in VE, but only the frequency of acentric fragments and the percentage of aberrant cells (excluding gaps) was significantly increased. No significant difference was found for the mean frequency of SCE. A statistically significant difference was also seen with the comet assay. The mean tail moment was higher in DM than in VE. However, no correlation was found between cytogenetic data and the effects in the comet assay. The results of our study indicate that DNA effects in the comet assay represent an independent endpoint which might be useful for the biomonitoring of genotoxic effects in addition to established tests.

Adult↗

Health effects among employees in sewage treatment plants: A literature survey.

BACKGROUND: Over the years, there have been regular reports in the media of work-related symptoms among employees at sewage treatment plants. Concern has spread among employees over the lack of knowledge of risk agents' symptoms, and diseases in that environment. This paper reviews the investigations of health risks in employees working in the sewage treatment plants. METHODS: A literature search was performed with the search terms; sewage, waste water, health effects, infection, and health hazards. Articles on health effects in relation to sewage were selected. RESULTS: Work in sewage water plants can involve exposure to different types of microorganisms and chemicals. The bacterial exposure is dominated by bacteria that naturally occur in nature. However, different bacteria and viruses that give rise to infections can be present in this environment and thus there exists a risk of infection, especially of hepatitis A. Investigations suggest that gastrointestinal tract symptoms are more common among employees at sewage treatment plants than among controls. Respiratory symptoms, fatigue, and headache have also been reported in several investigations. The cause of the symptoms is unknown, although certain data suggest that they are caused by inflammation. The results suggest that endotoxin in Gram-negative bacteria may be one of the causative agents. As regards cancer, some studies report an increased risk of stomach cancer and a few studies report an increased risk of cancer in the larynx, liver or, prostate or of leukemia. The spread of the cancers over a multitude of organs does not support a hypothesis of causality with agents commonly found in sewage treatment plants. CONCLUSIONS: Further investigations are needed to determine the work-related effects and ascertain the causal agents.

Cause of Death↗

Dynamical model development and parameter identification for an anaerobic wastewater treatment process.

This paper deals with the development and the parameter identification of an anaerobic digestion process model. A two-step (acidogenesis-methanization) mass-balance model has been considered. The model incorporates electrochemical equilibria in order to include the alkalinity, which has to play a central role in the related monitoring and control strategy of a treatment plant. The identification is based on a set of dynamical experiments designed to cover a wide spectrum of operating conditions that are likely to take place in the practical operation of the plant. A step by step identification procedure to estimate the model parameters is presented. The results of 70 days of experiments in a 1-m(3) fermenter are then used to validate the model.

Bacteria, Anaerobic↗

Maximization of acetic acid production in partial acidogenesis of swine wastewater.

Swine wastewater was biologically treated to produce short-chain volatile organic acids (VOAs) in laboratory-scale continuously stirred tank reactors. The maximum production rates of acetic and butyric acids associated with simultaneous changes in pH and hydraulic retention time (HRT) were investigated, in which the degree of acidification of swine wastewater to the short-chain VOAs was <25% of influent chemical oxygen demand (COD) concentration. A constant inoculum system was used to minimize the experimental error due to the use of inconsistent inoculum. The inoculum system was operated with synthetic wastewater at 6000 mg soluble chemical oxygen demand per liter (pH 6.0) and 35 degrees C at 0.5 day hydraulic retention time. Response surface methodology was applied successfully to determine the optimum physiological condition for which the maximum rate of acetic acid production occurred, which was pH 5.90 and 0.88 day hydraulic retention time at 35 degrees C. The partial acidification process to manage swine waste should be operated in the optimum condition for acetic acid production because the optimum operating condition for butyric acid production approached the washout point.

Acetic Acid↗

Bioreduction and biocrystallization of palladium by Desulfovibrio desulfuricans NCIMB 8307.

The reduction of Pd(II) to Pd(0) was accelerated by using the sulfate-reducing bacterium Desulfovibrio desulfuricans NCIMB 8307 at the expense of formate or H(2) as electron donors at pH 2-7. With formate no reduction occurred at pH 2, but with H(2) 50% of the activity was retained at pH 2, with the maximum rate (1.3-1.4 micromol min(-1) mg dry cells(-1)) seen at pH 3-7, which was similar to the rate with formate at neutral pH. Excess nitrate was inhibitory to Pd(II) reduction using formate, but not H(2). Chloride ion was inhibitory as low as 100 mM using formate but with H(2) only ca. 25% inhibition was observed at 500 mM Cl(-) and H(2) was concluded to be the electron donor of choice for the potential remediation of industrial wastes. Deposited Pd was visible on the cells using transmission and scanning electron microscopy and analysis by energy dispersive X-ray microanalysis (EDAX) identified the deposit as Pd, confirmed as Pd(0) by X-ray powder diffraction analysis (XRD). The crystal size of the biodeposited Pd(0) was determined to be only 50% of the size of Pd(0) crystals manufactured chemically from Pd(II) at the expense of H(2) and, unlike the chemically manufactured material, the biocrystal size was independent of the pH. The "biological" Pd(0) functioned as a superior chemical catalyst in a test reaction which liberated hydrogen from hypophosphite. Pd, and also Pt and Rh, could be recovered by resting cell suspensions under H(2) from an industrial processing wastewater, suggesting a possible future application of bioprocessing technology for precious metals.

Catalysis↗

Glutamic acid removal and PHB storage in the activated sludge process under dynamic conditions.

Glutamic acid removal in the activated sludge process is studied herein, primarily the formation of storage polymers under dynamic conditions. The activated sludge process was operated by using a sequencing batch reactor (sludge age of 6 d) fed with a synthetic mixture of readily available carbon sources, including glutamic acid. Removal of glutamic acid as the only carbon sources was studied in batch tests, along with oxygen consumption, ammonia uptake-release, and formation of storage polymers. It was found that poly-3-hydroxybutyrate (PHB) was stored and that the storage also occurred simultaneously to biomass growth. PHB storage accounted for 16% of the overall solids that were formed from glutamic acid, as the average value of nine batch tests. Neither other Polyhydroxyalkanoates nor polyglutamic acid were detected. Nuclear magnetic resonance analysis, performed on biomass extracts, allowed us to clarify the main metabolic pathways involved in glutamic acid removal and, in particular, the pathways involved in PHB storage. It was found that glutamic acid enters the Krebs cycle as alpha-ketoglutaric acid and exits to form pyruvic acid and then acetyl-CoA, which is the starting point of PHB production pathway.

Ammonia↗

Relationships between hydrodynamics and rheology of flocculating yeast suspensions in a high-cell-density airlift bioreactor.

In this article a hydrodynamic and rheological analysis of a continuous airlift bioreactor with high-cell-density system is presented. A highly flocculating recombinant strain of Sacharomyces cerevisiae containing genes for lactose transport (lactose permease) and hydrolysis (beta-galactosidase) was exploited to ferment lactose from cheese whey to ethanol. The magnetic particle-tracer method was used to assess the effect of operational conditions (air-flow rate, biomass concentration) on hydrodynamic behavior of an airlift bioreactor during the fermentation process. Measurements of liquid circulation velocity showed the existence of a critical value of biomass concentration at which a dramatic deceleration of net liquid flow appeared with increasing biomass quantity. Rheological analysis revealed exponential increase of viscosity of the yeast floc suspension at the same biomass concentration of about 73 g/dm3 corresponding to 42.8% v/v of solid fraction. These facts have a particular importance for the successful processing of a high-cell-density airlift bioreactor as only a circulated flow regime will be favorable to keep the solid particles in suspension state and evenly distributed throughout the bioreactor.

Bioreactors↗

Development and assessment of an innovative soil-washing process based on the use of cholic acid-derivatives as pollutant-mobilizing agents.

Surfactant-aided soil washing is often proposed for the restoration of aged organic pollutant-contaminated soils. As many of commercial surfactants have been found to be toxic and recalcitrant, the opportunity to use in this process cheap, non-toxic, and biodegradable pollutant-mobilizing agents, such as deoxycholic acid (DA), bovine bile (BB), and the residue resulting from DA extraction from BB (BBR), was studied in this work. A soil historically contaminated by chlorinated anilines and benzenes, thiophenes, and several polycyclic aromatic hydrocarbons was suspended at 15% w/v and washed in water or water amended at 1.0% (w/v) with DA, BB, BBR, or Triton X-100 (TX). The resulting effluents were supplemented with nutrients and subjected to aerobic bioremediation. The biogenic agents enhanced the water pollutant elution potential by 230/440%. TX enhanced the same parameter by about 540%; however, it mediated a lower depletion of the initial soil ecotoxicity and a more extensive mobilization of soil constituents with respect to the biogenic agents. Furthermore, TX adversely affected the biotreatability of resulting effluents, by adversely affecting the growth of cultivable bacterial biomass and the structure of eubacterial community of the effluent. On the contrary, the biogenic agents, and in particular DA and BB, enhanced the effluents bioremediation, by sustaining the growth and increasing the complexity of the effluent eubacterial communities. Thus, DA and BB are very promising additives for an effective and environmental friendly soil washing treatment of aged (chloro)organics contaminated soils.

Animals↗

Recovery of scrap iron metal value using biogenerated ferric iron.

The utility of employing biogenerated ferric iron as an oxidant for the recycling of scrap metal has been demonstrated using continuously growing cells of the extremophilic organism Acidithiobacillus ferrooxidans. A ferric iron rich (70 mol%) lixiviant resulting from bioreactor based growth of A. ferrooxidans readily solubilized target scrap metal with the resultant generation of a leachate containing elevated ferrous iron levels and solubilized copper previously resident in the scrap metal. Recovery of the copper value was easily accomplished via a cementation reaction and the clarified leachate containing a replenished level of ferrous iron as growth substrate was shown to support the growth of A. ferrooxidans and be fully recyclable. The described process for scrap metal recycling and copper recovery was shown to be efficient and economically attractive. Additionally, the utility of employing the E(h) of the growth medium as a means for monitoring fluctuations in cell density in cultures of A. ferrooxidans is demonstrated.

Acidithiobacillus↗

Event-driven time-optimal control for a class of discontinuous bioreactors.

Discontinuous bioreactors may be further optimized for processing inhibitory substrates using a convenient fed-batch mode. To do so the filling rate must be controlled in such a way as to push the reaction rate to its maximum value, by increasing the substrate concentration just up to the point where inhibition begins. However, an exact optimal controller requires measuring several variables (e.g., substrate concentrations in the feed and in the tank) and also good model knowledge (e.g., yield and kinetic parameters), requirements rarely satisfied in real applications. An environmentally important case, that exemplifies all these handicaps, is toxicant wastewater treatment. There the lack of online practical pollutant sensors may allow unforeseen high shock loads to be fed to the bioreactor, causing biomass inhibition that slows down the treatment process and, in extreme cases, even renders the biological process useless. In this work an event-driven time-optimal control (ED-TOC) is proposed to circumvent these limitations. We show how to detect a "there is inhibition" event by using some computable function of the available measurements. This event drives the ED-TOC to stop the filling. Later, by detecting the symmetric event, "there is no inhibition," the ED-TOC may restart the filling. A fill-react cycling then maintains the process safely hovering near its maximum reaction rate, allowing a robust and practically time-optimal operation of the bioreactor. An experimental study case of a wastewater treatment process application is presented. There the dissolved oxygen concentration was used to detect the events needed to drive the controller.

Biomass↗

Engineering TCE-degrading rhizobacteria for heavy metal accumulation and enhanced TCE degradation.

Many superfund sites are currently co-contaminated with organic pollutants such as trichloroethene (TCE) and heavy metals. A promising strategy to address these mixed-waste situations is the use of TCE-degrading rhizobacteria that will survive and thrive in soil heavily polluted with heavy metals. In this work, a gene coding for the metal-binding peptide, EC20, was introduced into rhizobacteria engineered for TCE degradation, resulting in strains with both metal accumulation and TCE degradation capabilities. EC20 was displayed onto the cell surface of Pseudomonas strain Pb2-1 and Rhizobium strain 10320D using an ice-nucleation protein (INP) anchor. Expression of EC20 was confirmed by Western blot analysis and cells with EC20 expression showed sixfold higher cadmium accumulation than non-engineered strains in the presence of 16 microM CdCl(2). As expected, the TCE degradation rate was reduced in the presence of cadmium for cells without EC20 expression. However, expression of EC20 (higher cadmium accumulation) completely restored the level of TCE degradation. These results demonstrated that EC20 expression enhanced not only cadmium accumulation but also reduced the toxic effect of cadmium on TCE degradation. We expect that similar improvements will be observed when these engineered rhizobacteria are inoculated onto plant roots.

Biodegradation, Environmental↗

Sorption of chromium(VI) from aqueous solution by cassava (Manihot sculenta Cranz.) waste biomass.

The sorption of highly toxic Cr(VI) ions by cassava waste biomass was quantitatively investigated. The sorption was found to be influenced by several physico-chemical factors such as agitation speed, temperature, contact time, pH, and sorbent/sorbate ratio. The adsorption data at equilibrium were fitted to Freundlich and Langmuir isotherms. The monolayer sorption capacity was found to be 61.79 mg of Cr(VI) per gram of biomass. The kinetics of Cr(VI) adsorption to pure cassava-tuber-bark wastes were determined based on a pseudo-second-order-rate model using the batch-sorption technique at a temperature of 30 degrees. The kinetics data suggest that the adsorption process is exothermic, and that the rate-limiting step is physisorption. Negative DeltaG(ads) values indicate that the adsorption is spontaneous and exothermic in nature. Also, under optimal conditions (in agitated 1M H(2)SO(4) at 30 degrees), the cassava waste biomass appears to be recyclable.

Absorption↗

Interlaboratory study for the validation of an ecotoxicological procedure to monitor the quality of septic sludge received at a wastewater treatment plant.

Septic tank sludge is regularly hauled to the Montreal Urban Community (MUC) wastewater treatment plant. It is then discharged and mixed with the wastewater inflow before entering the primary chemical treatment process. An ecotoxicological procedure integrating chemical and toxicological analyses has been recently developed and applied to screen for the illicit discharge of toxic substances in septic sludge. The toxicity tests used were the Microtox, the bacterial-respiration, and the lettuce (Lactuca sativa) root elongation tests. In order to validate the applicability of the proposed procedure, a two-year interlaboratory study was carried out. In general, the results obtained by two independent laboratories (MUC and the Centre d'expertise en analyse environnementale du Quebec) were comparable and reproducible. Some differences were found using the Microtox test. Organic (e.g., phenol and formaldehyde) and inorganic (e.g., nickel and cyanide) spiked septic sludge were detected with good reliability and high efficiency. The relative efficiency to detect spiked substances was > 70% and confirms the results of previous studies. In addition, the respiration test was the most efficient toxicological tool to detect spiked substances, whereas the Microtox was the least efficient (< 15%). Efficiencies to detect spiked contaminants were also similar for both laboratories. These results support previous data presented earlier and contribute to the validation of the ecotoxicological procedure used by the MUC to screen toxicity in septic sludge.

Environmental Monitoring↗

Polarography as a tool in peptide and protein analysis: studies on metal-chelating substances induced by cadmium in the algae Pheodactylum tricornutum and the graminae Agrostis capillaris.

Methods to determine characteristics and quantities of metal-binding substances are needed in order to identify phytochemical systems for use in agriculture and waste treatment. A metal-binding polypeptide (phytochelatin) was prepared by liquid chromatography from the algae Pheodactylum tricornutum and roots of the graminae Agrostis capillaris after exposure to cadmium. Polarographic (voltammetric) analysis indicated binding of cadmium by the polypeptide at different stabilities. No catalytic hydrogen peak at -1.7 V appeared in the polarogram of plant and algae metal-binding substance, but did appear in the polarogram of the metal-chelating protein metallothionein. In all polarograms, two significant peaks were detected. One at approximately -520 mV probably represents the reduction of single cadmium-sulfur bonds, and one at -930 mV the reduction of cadmium-thiolate complexes. A dose-response relationship between cadmium concentration and peak current at -930 mV was observed after exposure of roots of Agrostis at 5 and 10 microM.

Cadmium↗

Estimating the removal and biodegradation potential of radiolabeled organic chemicals in activated sludge.

A two-step procedure is described to characterize the removal and biodegradation potential of nonvolatile 14C-labeled organic compounds in activated sludge. In the first step, trace concentrations of radiolabeled test materials are dosed in influent wastewater to continuous-flow activated sludge (CAS) systems which have been previously exposed or acclimated to unlabeled test material. Radiolabel is quantified in influent, effluent, and activated sludge mixed liquor to determine total 14C removal and partitioning of radiolabel in solid and liquid compartments. The 14C data are used to calculate the amount of removal due to sorption and biodegradation and to estimate the apparent sorption coefficients for 14C activity to activated sludge solids. The 14C-labeled CAS studies are followed by biodegradation studies in batch-activated sludge (BAS) systems using sludge derived from the CAS system. The kinetics of biodegradation (defined as mineralization to 14CO2) are measured in the BAS system to confirm the CAS biodegradation results and generate mineralization rate constants for kinetic modeling. The two-step procedure was applied to radiolabeled anionic (linear alkylbenzene sulfonate) and cationic (dodecyltrimethylammonium chloride, distearyldimethylammonium chloride) surfactants which differed greatly in their biodegradation and sorption profiles. Laboratory removal figures for these materials were comparable to values measured in full-scale wastewater treatment systems, although the amount of removal due to sorption and biodegradation varied significantly for the different surfactants. In general, the 14C method has several advantages over standard methods used in the United States and Europe which employ unlabeled materials. These advantages include the use of realistic concentrations and test conditions for acclimating and dosing activated sludge microorganisms and the ability to generate partitioning and kinetic constants that can be used more broadly in environmental fate and exposure models.

Biodegradation, Environmental↗

Design and construction of wetlands for aqueous transfers and transformations of selected metals.

Two pilot-scale wetland cells (6.1 x 30.5 m) were integratively designed and constructed to emphasize and enhance transfers and transformations of selected metals (Cu, Pb, and Zn) in an aqueous matrix. A series of preliminary experiments and analyses were conducted to select macrofeatures (hydroperiod, hydrosoil, and vegetation) of the constructed wetland system. These wetland cells were designed to operate in series or parallel with nominal hydraulic retention times of 24-48 hr, respectively. With water at a depth of 30 cm, both wetland cells had hydrosoil (45 cm) planted with Scirpus californicus. After 250 days of wetland operation, average hydrosoil redox potentials in each wetland cell decreased from +90 mV to -165 mV, and average plant height increased from 0.3 to 2.7 m. Aqueous samples were collected over a 4.5-month period at the inflow and outflow sites of the wetland cells. Average inflow concentrations of total recoverable Cu, Pb, and Zn were 22.4, 10.5, and 565.9 micrograms/L, respectively. After a 46-hr HRT, average outflow concentrations of total recoverable Cu, Pb, and Zn were 15, 2.2, and 85.9 micrograms/L, resulting in removals of 33, 79, and 85%, respectively. Initial results suggest that these constructed wetlands can be designed to remove targeted metals in wastewater.

Industrial Waste↗

Biological degradation of cyclophosphamide and its occurrence in sewage water.

The mutagenic and cancerogenic antineoplastic agent cyclophosphamide (CP) is released into sewage water by cancer patient excretion. To assess the biological degradability of CP two standardized test systems, the Zahn-Wellens/EMPA test (OECD 302B) and a laboratory scale sewage treatment plant, were used. In both test systems the agent exhibited only poor degradability. To verify the expected occurrence of CP in hospital sewage, water samples were analyzed for CP with GC/MS after enrichment by solid-phase extraction. CP could be detected in concentrations ranging from 20 ng/L to 4.5 micrograms/L. The occurrence of the agent could also be proved in samples from the influent and the effluent of the communal sewage treatment plant into which the hospital's sewage water is shed. Concentrations ranged from 7 to 143 ng/L. In an attempt to assess the contribution of CP to the genotoxicity detected in hospital waste water in a recent study, the effects of CP in the umuC test, a bacterial genotoxicity assay, were investigated. However, no genotoxic effects of CP were found up to concentrations of 1 g/L.

Antineoplastic Agents, Alkylating↗

Environmental fate assessment of two synthetic polycarboxylate polymers.

In order to estimate the proportion which may enter the aquatic environment and characterize exposure potential, the environmental fate properties of two polycarboxylate polymers in activated sludge were determined. The polycarboxylates studied were a polymer emulsion (PE), a high-molecular-weight polymer (MW 50,000-60,000), and a resin polymer (RP), a low-molecular-weight polymer (MW 4500 to 9000). Both polymers exhibited a high degree of sorption to sludge solids in a series of sorption/ desorption studies, indicating a high potential for removal during wastewater treatment. In isotherm tests, the adsorption coefficient, Koc, was 1060 and 2730 for the RP and the PE, respectively. The biodegradation of the two polymers was measured under aerobic conditions in a 60-day semicontinuous activated sludge test and in a 60-day modified Sturm test using 14C-labeled compounds. In both studies, polymers accumulated in sludge solids over time and there was no evidence of significant biodegradation, although small amounts of radiolabeled CO2 were produced, suggesting the potential for biodegradation. Testing confirmed that the lack of biodegradation was not due to inhibition of microbial respiration by the polymers. High removals of PE and RP during wastewater treatment were confirmed in a field study at a facility receiving polymer inputs from households and manufacturing plants.

Biodegradation, Environmental↗