Biodegradation of hydrocarbon cuts used for diesel oil formulation

The biodegradability of various types of diesel oil (DO), such as straight-run DO, light-cycle DO, hydrocracking DO, Fischer–Tropsch DO and commercial DO, was investigated in biodegradation tests performed in closed-batch systems using two microflorae. The first microflora was an activated sludge from an urban wastewater treatment plant as commonly used in biodegradability tests of commercial products and the second was a microflora from a hydrocarbon-polluted soil with possible specific capacities for hydrocarbon degradation. Kinetics of CO₂ production and extent of DO biodegradation were obtained by chromatographic procedures. Under optimised conditions, the polluted-soil microflora was found to extensively degrade all the DO types tested, the degradation efficiencies being higher than 88%. For all the DOs tested, the biodegradation capacities of the soil microflora were significantly higher than those of the activated sludge. Using both microflora, the extent of biodegradation was highly dependent upon the type of DO used, especially its hydrocarbon composition. Linear alkanes were completely degraded in each test, whereas identifiable branched alkanes such as farnesane, pristane or phytane were degraded to variable extents. Among the aromatics, substituted mono-aromatics were also variably biodegraded.     

Application of the OECD 301F respirometric test for the biodegradability assessment of various potential endocrine disrupting chemicals

The biodegradability of several potential endocrine disrupting compounds, namely 4-n-nonylphenol (4-n-NP), nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), bisphenol A (BPA), triclosan (TCS), di-(2-ethylhexyl)-phthalate (DEHP), perfluorooctanoate (PFOA) and perfluorononanoate (PFNA) was evaluated in this study, using OECD method 301F (manometric respirometry test) and activated sludge as inoculum. According to the results, 4-n-NP and BPA meet the strict definition of ready biodegradability and they are not expected to be persistent during the activated sludge process. Partial biodegradation was observed for DEHP (58.7+/-5.7%, n=3), TCS (52.1+/-8.5%, n=3) and NP1EO (25.9+/-8.1%, n=3), indicating their possible biodegradation in wastewater treatment systems, while no biodegradation was observed for NP2EO, PFOA and PFNA. Experiments in the co-presence of a readily biodegradable compound showed the absence of co-metabolic phenomena during 4-n-NP, BPA and TCS biodegradation. Using first order kinetics to describe biodegradation of the target compounds, half-lives of 4.3+/-0.6, 1.3+/-0.2, 1.8+/-0.5, 6.9+/-2.6 days were calculated for 4-n-NP, BPA, TCS and DEHP, respectively. Toxicity tests using marine bacterium Vibrio fischeri showed that biodegradation of 4-n-NP, NP1EO, BPA and TCS is a simultaneous detoxification process, while possible abiotic or biotic transformations of NP2EO, DEHP, PFOA and PFNA during respirometric test resulted to significant increase of their toxicities.     

Modeling of activated sludge wastewater treatment processes

The performance of an activated sludge wastewater treatment process consisting of an aeration tank and a secondary settler has been studied. A tanks-in-series model with backflow was used for mathematical modeling of the activated sludge wastewater treatment process. Non-linear algebraic equations obtained from the material balances of MLSS (mixed liquor suspended solids or activated sludge), BOD (biological oxygen demand) and DO (dissolved oxygen) for the aeration tank and the settler and from the behavior of the settler were solved simultaneously using the modified Newton-Raphson technique. The concentration profiles of MLSS, BOD and DO in the aeration tank were obtained. The simulation results were examined from the viewpoints of mixing in the aeration tank and flow in the secondary settling tank. The relationships between the overall performance of the activated sludge process and the operating and design parameters such as hydraulic residence time, influent BOD, recycle ratio and waste sludge ratio were obtained.     

Interdependence between the aerobic degradation of BPA and readily biodegradable substrates by activated sludge in semi-continuous reactors

The objective of the present work was to analyze the interrelationship between the aerobic degradation of BPA and readily biodegradable substrates by activated sludge (AS) in semi-continuous reactors (SCRs). AS were obtained from three SCRs fed with glucose, acetate or peptone. AS from these reactors were used as inocula for three SCRs that were fed with each biogenic substrate, and for three SCRs that were fed with the biogenic substrate and BPA. In all cases, dissolved organic carbon (DOC), BPA, total suspended solids (TSS) and respirometric measurements were performed. Although BPA could be removed in the presence of all the tested substrates, AS grown on acetate exhibited the longest acclimation to BPA. Reactors fed with peptone attained the lowest TSS concentration; however, these AS had the highest specific BPA degradation rate. Specific DOC removal rates and respirometric measurements demonstrated that the presence of BPA had a negligible effect on the removal of the tested substrates. A mathematical model was developed to represent the evolution of TSS and DOC in the SCRs as a function of the operation cycle. Results suggest that the main effect of BPA on AS was to increase the generation of microbial soluble products. This work helps to understand the relationship between the biodegradation of BPA and readily biodegradable substrates.     

Production of acetone,butanol and ethanol from palm oil waste by Clostridium saccharoperbutylacetonicum N1-4

Summary A study of the feasibility of utilizing palm oil waste as a substrate for acetonebutanol-ethanol fermentation by Clostridium saccharoperbutylacetonicum N1-4 and, hence, to reduce the biochemical oxygen demand (BOD) of waste disposal was carried out. Among the two types of palm oil waste tested, separator sludge was the better substrate and it was able to support production of solvents by C. saccharoperbutylacetonicum N1-4 without a need for any mineral supplements. Enzymatic hydrolysis catalyzed by cellulase prior to fermentation was found to increase the yield of butanol by 75% (from 2.47g/l to 4.37g/l) and decreased the BOD by 66% (from 26,500 ppm to 8,900 ppm).     

Development and Evaluation of an Online CO2 Evolution Test and a Multicomponent Biodegradation Test System

Well-established biodegradation tests use biogenously evolved carbon dioxide (CO₂) as an analytical parameter to determine the ultimate biodegradability of substances. A newly developed analytical technique based on the continuous online measurement of conductivity showed its suitability over other techniques. It could be demonstrated that the method met all criteria of established biodegradation tests, gave continuous biodegradation curves, and was more reliable than other tests. In parallel experiments, only small variations in the biodegradation pattern occurred. When comparing the new online CO₂ method with existing CO₂ evolution tests, growth rates and lag periods were similar and only the final degree of biodegradation of aniline was slightly lower. A further test development was the unification and parallel measurement of all three important summary parameters for biodegradation—i.e., CO₂ evolution, determination of the biochemical oxygen demand (BOD), and removal of dissolved organic carbon (DOC)—in a multicomponent biodegradation test system (MCBTS). The practicability of this test method was demonstrated with aniline. This test system had advantages for poorly water-soluble and highly volatile compounds and allowed the determination of the carbon fraction integrated into biomass (heterotrophic yield). The integrated online measurements of CO₂ and BOD systems produced continuous degradation curves, which better met the stringent criteria of ready biodegradability (60% biodegradation in a 10-day window). Furthermore the data could be used to calculate maximal growth rates for the modeling of biodegradation processes.     

Influence of methanol/sorbitol co-feeding rate on pAOX1 induction in a <Emphasis Type="Italic">Pichia pastoris</Emphasis> Mut+ strain in bioreactor with limited oxygen transfer rate

High Pichia pastoris biomass density could be obtained using high co-feeding rate of methanol and sorbitol in a fed-batch or continuous culture, while further higher feeding rate finally leads to oxygen limitation in bioreactor. In the literature, there is lack of report about AOX1 promoter regulation with regard to dissolved oxygen level (DO). Therefore, in this work, chemostat cultures were performed to investigate the cell growth, metabolism and regulation of the AOX1 promoter (pAOX1) regarding co-feeding rate of optimized methanol/sorbitol mixture (methanol fraction 0.60 C-mol/C-mol) using a P. pastoris Mut+/pAOX1-lacZ strain. The oxygen transfer rates (OTR) in bioreactor were kept in the range of typical values of large bioreactor, i.e., 4–8 g/(L h) if DO equals 30 % saturation or 5–10 g/(L h) if DO nears zero. For DO >0, an increase of the carbon fed led to an increase of pAOX1 induction. By contrast, when dissolved oxygen was completely depleted, methanol accumulated, causing a 30 % decrease of pAOX1 induction. However, this decrease is more likely to be lined to methanol accumulation than to low level of dissolved oxygen (<4 % DO). Methanol/sorbitol co-feeding allowed cells to adapt to oxygen transient limitations that often occur at industrial scale with reduced effect on pAOX1 induction. The optimal feeding rate tested here was 6.6 mmol C (DCW h)^?1 at an OTR of 8.28 g O₂(L h)^?1 with over fivefold pAOX1 induction (probably directly associated with target protein productivity) compared with previous work.     

Degradation of bisphenol A and nonylphenol by nitrifying activated sludge

Nitrifier-enriched activated sludge was used in batch degradation of bisphenol A (BPA) and nonylphenol (NP) to verify if ammonium-oxidizing activity within nitrifiers is responsible for the biodegradation of those compounds. Decreases in both BPA and NP concentrations occurred simultaneously with ammonium (NH₄⁺) oxidation into nitrate (NO₃⁻) by nitrifying sludge. However, when ammonium was replaced by nitrite (NO₂⁻) in the medium, an acclimation period was required prior onset of significant degradation of BPA and NP. In the presence of inhibitors such as allylthiourea or Hg₂SO₄, BPA and/or NP reduction decreased significantly, implying that removal of BPA and NP was mostly mediated by biological activity rather than by physicochemical adsorption onto sludge flocs.     

Comparison of experimental methods for determination of toxicity and biodegradability of xenobiotic compounds

Different methods for determining the toxicity and biodegradability of hazardous compounds evaluating their susceptibility to biological treatment were studied. Several compounds including chlorophenols and herbicides have been evaluated. Toxicity was analyzed in terms of EC₅₀ and by a simple respirometric procedure based on the OECD Method 209 and by the Microtox^® bioassay. The values of EC₅₀ obtained from respirometry were in all the cases higher than those from the Microtox^® test. The respirometric inhibition values of chlorophenols were related well with the number of chlorine atoms and their position in the aromatic ring. In general, herbicides showed lower inhibition, being alachlor the less toxic from this criterion. For determination of biodegradability an easier and faster alternative to the OECD Method 301, with a higher biomass to substrate ratio is proposed. When this test was negative, the Zahn-Wellens one was performed in order to evaluate the inherent biodegradability. In the fast test of biodegradability, 4-chlorocatechol and 4-chlorophenol showed a complete biodegradation by an unacclimated sludge upon 48 h. These results together with their low respirometric inhibition, allow concluding that these compounds could be conveniently removed in a WWTP. Alachlor, 2,4-dichlorophenol, 2,4,6-trichlorophenol and MCPA showed a partial biodegradation upon 28 days by the Zahn-Wellens inherent biodegradability test.     

A rational approach for predicting the dissolved oxygen profile in receiving waters

A practical method for predicting the dissolved oxygen (DO) profile in a stream receiving biodegradable waste is presented. In this method the BOD (accumulated O₂ uptake) curve is obtained using an open jug technique. The accumulated O₂ uptake curve thus developed is employed in numerical integration with the physical reaeration data for the receiving stream to predict the DO profile in the stream. In the present study, the method was examined using 10-liter open jar reactors to obtain the O₂ uptake curves, and the receiving stream was a 670-liter simulated stream apparatus which has been employed in previous studies on stream reaeration. The method was found to provide a fairly good prediction of the actual dissolved oxygen profile observed in the receiving stream. The effect of the reaeration constant, K₂ )agitation effect(, on the kinetics of O₂ uptake was also investigated and it was found that increased agitation (higher K₂ value) caused some increase in the accumulated oxygen uptake (BOD) curve with most of the increase coming after the so-called “plateau” area in the O₂ uptake curve, i.e., in the general case, after the low point along the DO sag curve.     

A flow injection analysis system with encapsulated high-density <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> cells for rapid determination of biochemical oxygen demand

The biochemical oxygen demand (BOD) determination was studied using a novel flow injection analysis (FIA) system with encapsulated Saccharomyces cerevisiae cells and an oxygen electrode and was compared with conventional 5-day BOD tests. S. cerevisiae cells were packed in a calcium alginate capsule at a dry cell weight of 250 g/l of capsule core. The level of dissolved oxygen (DO) was reduced due to the enhanced respiratory activity of the microbial cells when the injected nutrient passed through the bioreactor. The decrease in DO (ΔDO) was intensified with the amount of microbial cells packed in the bioreactor. However, the specific ΔDO decreased as the amount of cells loaded in the bioreactor increased. The ΔDO value was dependent on the pH and temperature of the mobile phase and reached its maximum value at 35°C and pH 7–8. Also, ΔDO became larger at longer response times as the flow rate of the mobile phase decreased. The measurement of ΔDO was repeated more than six times consecutively using a 20-ppm standard glucose and glutamic acid solution, which confirmed the reproducibility with a standard deviation of 0.95%. A strong linear correlation between ΔDO and BOD was also observed. The 5-day BOD values of actual water and wastewater samples were in accordance with the BOD values obtained by this FIA method using encapsulated S. cerevisiae cells. Unlike the cell-immobilized bead system, there was no contamination of the bioreactor resulting from any leak of yeast cells from the sensor capsules during BOD measurements.     

Relationship between respirometric activity and community of entrapped nitrifying bacteria: Implications for partial nitrification

Activated sludge obtained from two municipal wastewater treatment facilities (WWTF) was used as seed sludge for enriched nitrifiers, which were later entrapped in polyvinyl alcohol. Seed sludge from one WWTF was acclimated to high ammonia level (1813 mg NH₃-N l^?1) through the return of sludge digester supernatant back to primary clarifier while seed sludge from the other WWTF was un-acclimated. To elucidate on how to control partial nitrification by entrapped cells, which could be different from suspended cells, kinetics of entrapped enriched nitrifiers were studied using a respirometric assay. The community of nitrifiers within the entrapment matrix, which was observed by fluorescence in situ hybridization (FISH) technique, was related to the nitritation and nitratation kinetics based on oxygen uptake rate. Maximum oxygen uptake rate, and substrate and oxygen affinities of both ammonia oxidizing bacteria (AOB) for nitritation and nitrite oxidizing bacteria (NOB) for nitratation in entrapped cells were lower than those of corresponding suspended cells. Under dissolved oxygen (DO) limiting conditions, nitratation was more suppressed than nitritation for suspended cells, while for the entrapped cells, the results were the contrary. A free ammonia (FA) inhibition affected only the un-acclimated sludge. Either FA inhibition or DO limitation might not be a sole effective control parameter to achieve partial nitrification by entrapped cells. FISH results revealed that Nitrosomonas europaea was the dominant AOB while Nitrobacter species was the dominant NOB in all cases. Heterotrophs were also present in the entrapment at 22.8 ± 18.6% and 41.5 ± 4.3% of total bacteria for acclimated and un-acclimated originated sludge. The availability of substrate and oxygen governed the distributions of AOB, NOB and heterotrophs within the entrapment and nitritation kinetics of entrapped nitrifiers.     

UASB/Flash aeration enable complete treatment of municipal wastewater for reuse

A simple, efficient and cost-effective method for municipal wastewater treatment is examined in this paper. The municipal wastewater is treated using an upflow anaerobic sludge bed (UASB) reactor followed by flash aeration (FA) as the post-treatment, without implementing aerobic biological processes. The UASB reactor was operated without recycle, at hydraulic retention time (HRT) of 8 h and achieved consistent removal of BOD, COD and TSS of 60-70% for more than 12 months. The effect of FA on UASB effluent post-treatment was studied at different HRT (15, 30 and 60 min) and dissolved oxygen (DO) concentrations (low DO = 1-2 mg/L and high DO = 5-6 mg/L). The optimum conditions for BOD, COD and sulfide removal were 30-60 min HRT and high DO concentration inside the FA tank. The final effluent after clarification was characterized by BOD and COD values of 28-35 and 50-58 mg/L, respectively. Sulfides were removed by more than 80%, but the fecal coliform only by ~2 log. The UASB followed by FA is a simple and efficient process for municipal wastewater treatment, except for fecal coliform, enabling water and nutrients recycling to agriculture.     

Comparison of oxygen consumption in freshwater mussels (Unionidae) from different habitats during declining dissolved oxygen concentration

The rate of oxygen consumption (OC) of 9 species of freshwater mussels was measured under declining dissolved oxygen (DO) concentrations. The effects of temperature for some species also was investigated. The pattern of the OC vs. DO curve for each species was used in a hyperbolic model to compare abilities to regulate OC under low oxygen conditions. At 24.5 °C, Pyganodon grandis (from lakes), Amblema plicata and Quadrula pustulosa (from mud or sand in large rivers), Elliptio complanata (from pool areas in rivers), and Elliptio fisheriana and Elliptio lanceolata (from bank margins of rivers) were better able to maintain OC under low DO than were Villosa iris and Villosa constricta (which inhabit riffles) and Pleurobema cordatum (found in rivers with moderate flow). Villosa iris was especially sensitive to low oxygen conditions. The ability to maintain normal OC at low DO was improved considerably at 16.5 °C for V. iris, P. grandis and E. complanata. It is concluded that oxygen regulation ability appears to be related to the degree of hypoxia a species normally experiences in its habitat type, and it is enhanced at low temperature. The measurement of OC vs. DO may be a useful technique for estimating DO water quality criteria for endangered species because it is noninvasive.     

Survey of the Anaerobic Biodegradation Potential of Organic Chemicals in Digesting Sludge

The degradation potential of 77 organic chemicals under methanogenic conditions was examined with an anaerobic digesting sludge from the United Kingdom. Degradation was assessed in terms of net total gas (CH₄ plus CO₂) produced, expressed as a percentage of the theoretical production (ThGP). The compounds tested were selected from various chemical groups and included substituted phenols and benzoates, pesticides, phthalic acid esters, homocyclic and heterocyclic ring compounds, glycols, and monosubstituted benzenes. The results obtained were in good agreement with published surveys of biodegradability in U.S. digesting sludges and other methanogenic environments. In general, the presence of chloro or nitro groups inhibited anaerobic gas production, while carboxyl and hydroxyl groups facilitated biodegradation. The relationship between substituent position and susceptibility to methanogenic degradation was compound dependent. The following chemicals were completely degraded (≥80% ThGP) at a concentration of 50 mg of carbon per liter: phenol, 2-aminophenol, 4-cresol, catechol, sodium benzoate, 4-aminobenzoic acid, 3-chlorobenzoic acid, phthalic acid, ethylene glycol, diethylene glycol, triethylene glycol, sodium stearate, and quinoline. 3-Cresol, 4-chlorobenzoic acid, dimethyl phthalate, and pyridine were partially degraded. Although the remaining chemicals tested were either persistent or toxic, their behavior may differ at more environmentally realistic chemical-to-biomass ratios. Our findings suggest that biodegradability assessments made with sludge from one source can be extrapolated to sludge from another source with a reasonable degree of confidence and should help in predicting the fate of an organic chemical during the anaerobic digestion of sewage sludge.     

Comparison of biological removal via nitrite with real-time control using aerobic granular sludge and flocculent activated sludge

The process of nitrification–denitrification via nitrite for nitrogen removal under real-time control mode was tested in two laboratory-scale sequencing batch reactors (SBRs) with flocculent activated sludge (R1) and aerobic granular sludge (R2) to compare operational performance and real-time control strategies. The results showed that the average ammonia nitrogen, total inorganic nitrogen (TIN), and chemical oxygen demand (COD) removal during aeration phase were 97.6%, 57.0%, and 90.1% in R2 compared with 98.6%, 48.7%, and 88.1% in R1. The TIN removed in both SBRs was partially due to the presence of simultaneous nitrification–denitrification via nitrite, especially in R2. The specific nitrification and denitrification rates in R2 were 0.0416 mgNH₄⁺–N/gSS-min and 0.1889 mgNO_X⁻–N/gSS-min, which were 1.48 times and 1.35 times that of R1. The higher rates for COD removal, nitrification, and denitrification were achieved in R2 than R1 with similar influent quality. Dissolved oxygen (DO), pH, and oxidization reduction potential, corresponding to nutrient variations, were used as diagnostic parameters to control the organic carbon degradation and nitrification–denitrification via nitrite processes in both SBRs. The online control strategy of granular SBR was similar to that of the SBR with flocculent activated sludge. However, a unique U-type pattern on the DO curve in granular SBR was different from SBR with flocculent activated sludge in aerobic phase.     

Effects of dilution on dissolved oxygen depletion and microbial populations in the biochemical oxygen demand determination

The biochemical oxygen demand (BOD) value is still a key parameter that can determine the level of organics, particularly the content of biodegradable organics in water. In this work, the effects of sample dilution, which should be done inevitably to get appropriate dissolved oxygen (DO) depletion, on the measurement of 5-day BOD (BOD₅), was investigated with and without seeding using natural and synthetic water. The dilution effects were also evaluated for water samples taken in different seasons such as summer and winter because water temperature can cause a change in the types of microbial species, thus leading to different oxygen depletion profiles during BOD testing. The predation phenomenon between microbial cells was found to be dependent on the inorganic nutrients and carbon sources, showing a change in cell populations according to cell size after 5-day incubation. The dilution of water samples for BOD determination was linked to changes in the environment for microbial growth such as nutrition. The predation phenomenon between microbial cells was more important with less dilution. BOD₅ increased with the specific amount of inorganic nutrient per microbial mass when the natural water was diluted. When seeding was done for synthetic water samples, the seed volume also affected BOD due to the rate of organic uptake by microbes. BOD₅ increased with the specific bacterial population per organic source supplied at the beginning of BOD measurement. For more accurate BOD measurements, specific guidelines on dilution should be established.     

Continuous fill intermittent decant type sequencing batch reactor application to upgrade the UASB treated sewage

The performance of continuous flow intermittent decant type sequencing batch (CFID) reactor treating the effluent of an UASB reactor treating domestic wastewater and operated at 8 h hydraulic retention time (HRT) was investigated. The CFID was operated at three different HRTs (22, 8 and 6 h) and three different dissolved oxygen (DO) patterns (<0.5, 2.5–3.5 and 3.5–4.5 mg/L). The highest effluent quality was observed at the 8 h HRT and 2.5–3.5 mg/L DO concentration. At this operational condition, the average BOD, TSS, ammonia nitrogen and fecal coliform removal efficiencies were 83, 90, 74 and 99 %, respectively. The CFID is a promising post-treatment option for existing UASB systems, with a final effluent quality that comply with receiving water and effluent reuse criteria.     

Towards an on-line biochemical oxygen demand analyser

A simple, and effective tool to measure BOD with a 98% correlation to theoretical BOD values has been developed using a technique of continuous short term measurements of O₂ consumption rates. Measurement times are in the range of 5 to 10 h depending on the initial BOD, instead of 5 days required in the conventional BOD₅ analysis. A bioreactor containing calcium alginate entrapped activated sludge microorganisms was used with phenol and hydroxybutyric acid (HBA) as model substrates.     

Analysis of factors affecting the performance of partial nitrification in a sequencing batch reactor

The objective of this study was to analyze the factors affecting the performance of partial nitrification in a sequencing batch reactor. During a 140-day long-term operation, influent pH value, dissolved oxygen (DO), and chemical oxygen demand/nitrogen (COD/N) ratio were selected as operating factors to evaluate the maintenance and recovery of nitrite accumulation. Results showed that high DO concentration (2–4 mg/L) could damage nitrite accumulation immediately. However, nitrite accumulation ratio (NAR) could be increased from 1.68?±?1.51 to 35.46?±?7.86 % when increasing the pH values from 7.5 to 8.3 due to the increased free ammonia concentration. Afterwards, stable partial nitrification and high NAR could be recovered when the reactor operated under low DO concentration (0.5–1.0 mg/L). However, it required a long time to recover the partial nitrification of the reactor when the influent COD/N ratios were altered. Fluorescence in situ hybridization analysis implied that ammonium oxidizing bacteria were completely recovered to the dominant nitrifying bacteria in the system. Meanwhile, sludge volumetric index of the reactor gradually decreased from 115.6 to 56.6 mL/g, while the mean diameter of sludge improved from74.57 to 428.8 μm by using the strategy of reducing settling time. The obtained results could provide useful information between the operational conditions and the performance of partial nitrification when treating nitrogen-rich industrial wastewater.