BOD biosensors for pulp and paper industry wastewater analysis

Introduction

Two semi-specific microbial biosensors were constructed for the analysis of biochemical oxygen demand (BOD) in high-cellulose-content pulp and paper industry wastewaters. The biosensors were based on living cells of Bacillus subtilis and Paenibacillus sp. immobilized in an agarose gel matrix. Semi-specific microorganisms were isolated from various samples (decaying sawdust and rabbit manure) and were chosen based on their ability to assimilate cellulose.

Materials &; methods

The biosensors were calibrated with the Organization for Economic Cooperation and Development synthetic wastewater, and measurements with different wastewaters were conducted.

Results

The response time of biosensors using the steady-state method was 20?C25 min, and the service life of immobilized microorganisms was 96 days. Detection limit was 5 mg/l of BOD₇ while linear ranges extended up to 55 and 50 mg/l of the BOD₇ for B. subtilis- and Paenibacillus sp.-based biosensors, respectively. Repeatability and reproducibility of both biosensors were within the limits set by APHA??less than 15.4%. In comparison, both biosensors overestimated the BOD₇ values in paper mill wastewaters and underestimated the BOD₇ in aspen pulp mill wastewater.

Conclusions

The semi-specific biosensors are suitable for the estimation of organic pollution derived from cellulose, while the detection of pollution derived from tannins and lignins was minor. Better results in terms of accuracy and repeatability were gained with Paenibacillus sp. biosensor.     

A comparative analysis of the characteristics of a range of real and synthetic wastewaters

Synthetic wastewaters are widely used in many fields of wastewater research and operational management. However, few comparative studies have been conducted on the large number of published formulations. Eleven synthetic formulations simulating municipal wastewaters were selected based on their frequency of use, relative complexity, ease of formulation and cost and compared to two real municipal wastewaters. Synthetic wastewaters vary significantly in their compositions and characteristics, especially in terms of alkalinity, BOD K ₁, SOUR, BOD_U, COD/BOD and C/N/P ratio, although they are portrayed as ‘typical’ in terms of characteristics and suitability for use. The pH, alkalinity and the presence of Ca and Mg should be considered in combination with the diluent used. Where the diluent is tap water, then the presence of treatment chemicals should also be considered. The effects of the micronutrients present are also an important factor. The study found that no single formulation is appropriate for all situations. Both the Syntho and Synthes formulations attempt to simulate real wastewater, whereas other formulations primarily act as readily biodegradable vectors for toxicity analyses, characterisation studies and treatment process evaluations. The criteria for choosing a particular synthetic wastewater very much depend on its intended application and require careful selection.     

Bioelectrochemical anaerobic sewage treatment technology for Arctic communities

This study describes a novel wastewater treatment technology suitable for small remote northern communities. The technology is based on an enhanced biodegradation of organic carbon through a combination of anaerobic methanogenic and microbial electrochemical (bioelectrochemical) degradation processes leading to biomethane production. The microbial electrochemical degradation is achieved in a membraneless flow-through bioanode–biocathode setup operating at an applied voltage below the water electrolysis threshold. Laboratory wastewater treatment tests conducted through a broad range of mesophilic and psychrophilic temperatures (5–23 °C) using synthetic wastewater showed a biochemical oxygen demand (BOD₅) removal efficiency of 90–97% and an effluent BOD₅ concentration as low as 7 mg L^?1. An electricity consumption of 0.6 kWh kg^?1 of chemical oxygen demand (COD) removed was observed. Low energy consumption coupled with enhanced methane production led to a net positive energy balance in the bioelectrochemical treatment system.     

Synthetic olive mill wastewater treatment by Fenton’s process in batch and continuous reactors operation

Degradation of total phenol (TPh) and organic matter, (expressed as total organic carbon TOC), of a simulated olive mill wastewater was evaluated by the Fenton oxidation process under batch and continuous mode conditions. A mixture of six phenolic acids usually found in these agro-industrial wastewaters was used for this purpose. The study focused on the optimization of key operational parameters of the Fenton process in a batch reactor, namely Fe²⁺ dosage, hydrogen peroxide concentration, pH, and reaction temperature. On the assessment of the process efficiency, > 99% of TPh and > 56% of TOC removal were attained when [Fe²⁺] = 100 ppm, [H₂O₂] = 2.0 g/L, T = 30 °C, and initial pH = 5.0, after 300 min of reaction. Under those operational conditions, experiments on a continuous stirred-tank reactor (CSTR) were performed for different space-time values (τ). TOC and TPh removals of 47.5 and 96.9%, respectively, were reached at steady-state (for τ = 120 min). High removal of COD (> 75%) and BOD₅ (> 70%) was achieved for both batch and CSTR optimum conditions; analysis of the BOD₅/COD ratio also revealed an increase in the effluent’s biodegradability. Despite the high removal of lumped parameters, the treated effluent did not met the Portuguese legal limits for direct discharge of wastewaters into water bodies, which indicates that coupled chemical-biological process may be the best solution for real olive mill wastewater treatment.     

Flow-rate and pollution characteristics of domestic wastewater

The characteristics of the flow-rate and pollution parameters of the domestic wastewater of a medium-size city (Sivas, Turkey) have been investigated. The design and the operating parameters of a wastewater treatment plant appropriate to such a settlement were also studied. Daily and monthly flow-rate measurements and wastewater samples were taken at the outlet. Peak values of the flow-rate were observed between the hours 09:00 and 14:00. Domestic wastewater production per capita was determined as 170.6 litres per day. The wastewater biological reaction ratio coefficient (k), the BOD₅/COD ratio and the BOD₅/N ratio varied in the ranges 0.113–0.312, 0.44–0.67 and 3.1–9.5 per day, respectively. Daily pollution load parameters per capita were determined as follows: 47.3 g for BOD₅, 85.3 g for COD, 34.9 g for suspended solids, 7.9 g for nitrogen and 1.32 g for phosphorus.     

Applying Kohonen self-organizing map as a software sensor to predict biochemical oxygen demand.

The 5 days at 20 degrees C biochemical oxygen demand (BOD5) is an important parameter for monitoring organic pollution in water and assessing the biotreatability of wastewater. Moreover, BOD5 is used for wastewater treatment plant discharge consents and other water pollution control purposes. However, the traditional bioassay method for estimating the BOD5 involves the incubation of sample water for 5 days. It follows that BOD5 is not available for real-time decisionmaking and process control purposes. On the other hand, previous efforts to solve this problem by developing more rapid biosensors had limited success. This paper reports on the development of Kohonen self-organizing map (KSOM)-based software sensors for the rapid prediction of BOD5. The findings indicate that the KSOM-based BOD5 estimates were in good agreement with those measured using the conventional bioassay method. This offers significant potential for more timely intervention and cost savings during problem diagnosis in water and wastewater treatment processes.     

A full-scale sequencing batch reactor system for swine wastewater treatment

A full-scale sequencing batch reactor (SBR) system was evaluated for its ability to remove carbon and nitrogen from swine wastewater. The SBR was operated on four, six-hour cycles each day, with each cycle consisting of 4.5 hours of “React,” 0.75 hours of “Settling”, 0.75 hours for “Draw” and “Fill.” Within each cycle, an amount of wastewater equivalent to about 5% of the reactor volume (5,500 litres) was removed and added. The SBR system was able to remove 82% of biochemical oxygen demand (BOD) and more than 75% of nitrogen. Even though the SBR effluent, with an average effluent BOD₅ of about 588 mg L^{? 1}, did not meet the discharge criteria, it enabled a reduction of the land base required for land application of swine wastewater by about 75%. Results indicated that the SBR system was a viable method for the treatment of swine wastewater.     

Health care industries: potential generators of genotoxic waste

Health care waste includes all the waste generated by health care establishments, research facilities, and laboratories. This constitutes a variety of chemical substances, such as pharmaceuticals, radionuclides, solvents, and disinfectants. Recently, scientists and environmentalists have discovered that wastewater produced by hospitals possesses toxic properties due to various toxic chemicals and pharmaceuticals capable of causing environmental impacts and even lethal effects to organisms in aquatic ecosystems. Many of these compounds resist normal wastewater treatment and end up in surface waters. Besides aquatic organisms, humans can be exposed through drinking water produced from contaminated surface water. Indeed, some of the substances found in wastewaters are genotoxic and are suspected to be potential contributors to certain cancers. The aim of this study was to evaluate the genotoxic and cytotoxic potential of wastewaters from two hospitals and three clinical diagnostic centers located in Jaipur (Rajasthan State), India using the prokaryotic Salmonella mutagenicity assay (Ames assay) and the eukaryotic Saccharomyces cerevisiae respiration inhibition assay. In the Ames assay, untreated wastewaters from both of the health care sectors resulted in significantly increased numbers of revertant colonies up to 1,000–4,050 as measured by the Salmonella typhimurium TA98 and TA100 strains (with and without metabolic activation) after exposure to undiluted samples, which indicated the highly genotoxic nature of these wastewaters. Furthermore, both hospital and diagnostic samples were found to be highly cytotoxic. Effective concentrations at which 20 % (EC₂₀) and 50 % (EC₅₀) inhibition of the respiration rate of the cells occurred ranged between ～0.00 and 0.52 % and between 0.005 and 41.30 % (calculated with the help of the MS excel software XLSTAT 2012.1.01; Addinsoft), respectively, as determined by the S. cerevisiae assay. The results indicated that hospital wastewaters contain genotoxic and cytotoxic components. In addition, diagnostic centers also represent small but significant sources of genotoxic and cytotoxic wastes.     

Integrated Bacillus sp. immobilized cell reactor and Synechocystis sp. algal reactor for the treatment of tannery wastewater

The wastewater discharged from leather industries lack biodegradability due to the presence of xenobiotic compounds. The primary clarification and aerobic treatment in Bacillus sp. immobilized Chemo Autotrophic Activated Carbon Oxidation (CAACO) reactor removed considerable amount of pollution parameters. The residual untreated organics in the wastewater was further treated in algal batch reactor inoculated with Synechocystis sp. Sodium nitrate, K₂HPO₄, MgSO₄.7H₂O, NH₄Cl, CaCl₂·2H₂O, FeCl₃ (anhydrous), and thiamine hydrochloride, rice husk based activated carbon (RHAC), immobilization of Bacillus sp. in mesoporous activated carbon, sand filter of dimensions diameter, 6 cm and height, 30 cm; and the CAACO reactor of dimensions diameter, 5.5 cm and height, 30 cm with total volume 720 ml, and working volume of 356 ml. In the present investigation, the CAACO treated tannery wastewater was applied to Synechocystis sp. inoculated algal batch reactor of hydraulic residence time 24 h. The BOD₅, COD, and TOC of treated wastewater from algal batch reactor were 20?±?7, 167?±?29, and 78?±?16 mg/l respectively. The integrated CAACO system and Algal batch reactor was operated for 30 days and they accomplished a cumulative removal of BOD₅,COD, TOC, VFA and sulphide as 98 %, 95 %, 93 %, 86 %, and 100 %, respectively. The biokinetic constants for the growth of algae in the batch reactor were specific growth rate, 0.095(day^?1) and yield coefficient, 3.15 mg of algal biomass/mg of COD destructed. The degradation of xenobiotic compounds in the algal batch reactor was confirmed through HPLC and FT-IR techniques. The integrated CAACO–Algal reactor system established a credible reduction in pollution parameters in the tannery wastewater. The removal mechanism is mainly due to co-metabolism between algae and bacterial species and the organics were completely metabolized rather than by adsorption.     

发酵类抗生素废水处理工艺及运行效果

介绍了内蒙古某抗生素废水处理厂的处理工艺流程、处理单元类型及其详细设计参数,通过对该污水处理厂半年内运行效果的调研,分析评价该污水处理系统的处理效果及出水指标;该水厂工艺流程对COD、TOC、氨氮、总氮、总磷、BOD和急性毒性的平均去除率分别为96.52%、95.83%、50.24%、55.20%、62.05%、98.52%和99.14%,COD、氨氮、总氮、总磷出水水质分别为477、438、556.7和7.69 mg/L,达到排放到城区下水道要求,园区内所有废水汇集到园区污水处理厂进行集中深度处理达标后排放水体。通过对该水厂工艺的介绍和水质分析,为我国抗生素生产企业面临的废水处理工艺类型选择及工程改造升级提供一定参考。     

Comparison of naphthalene bioavailability determined by whole-cell biosensing and availability determined by extraction with Tenax

A rapid biological method for the determination of the bioavailability of naphthalene was developed and its value as an alternative to extraction-based chemical approaches demonstrated. Genetically engineered whole-cell biosensors are used to determine bioavailable naphthalene and their responses compared with results from Tenax extraction and chemical analysis. Results show a 1:1 correlation between biosensor results and chemical analyses for naphthalene-contaminated model materials and sediments, but the biosensor assay is much faster. This work demonstrates that biosensor technology can perform as well as standard chemical methods, though with some advantages including the inherent biological relevance of the response, rapid response time, and potential for field deployment. A survey of results from this work and the literature shows that bioavailability under non-equilibrium conditions nonetheless correlates well with K_oc or K_d. A rationale is provided wherein chemical resistance is speculated to be operative.     

Variation of raw wastewater microbiological quality in dry and wet weather conditions

The microbiological quality of urban wastewaters presents important environmental, sanitary, and political challenges. However, the variability of untreated wastewater quality is seldom known when it comes to microbial parameters. This study aims to evaluate the variability of microbiological quality in wastewater influents from different wastewater treatment plants connected to combined and partially separate sewer networks in the Parisian area and to evaluate the impact of this variability on the treatment efficiency and on the accuracy of wastewater effluent monitoring. The densities of fecal indicator bacteria (FIB), Escherichia coli and intestinal enterococci, and their partitioning on settleable particles were analyzed at the inlet of two wastewater treatment plants during dry weather (130 composite samples and 7 days sampled every 2 hours) and storm events (39 composite samples, and 7 rain courses) from 2008 to 2012. The results showed that fecal indicator densities vary according to the network characteristics and according to the meteorological conditions. During storm events, a significant dilution of E. coli and enterococci was observed, as well as a decrease in the settleable fraction of E. coli during the maximal impact of the storm. However, storm events did not significantly impact the regular FIB monitoring. FIB removals by primary and secondary treatment were significantly correlated with FIB densities in influent wastewater; however, meteorological conditions also influenced the removal of FIB.     

Construction and comparison of fluorescence and bioluminescence bacterial biosensors for the detection of bioavailable toluene and related compounds

Environmental pollution with petroleum products such as benzene, toluene, ethylbenzene, and xylenes (BTEX) has garnered increasing awareness because of its serious consequences for human health and the environment. We have constructed toluene bacterial biosensors comprised of two reporter genes, gfp and luxCDABE, characterized by green fluorescence and luminescence, respectively, and compared their abilities to detect bioavailable toluene and related compounds. The bacterial luminescence biosensor allowed faster and more-sensitive detection of toluene; the fluorescence biosensor strain was much more stable and thus more applicable for long-term exposure. Both luminescence and fluorescence biosensors were field-tested to measure the relative bioavailability of BTEX in contaminated groundwater and soil samples. The estimated BTEX concentrations determined by the luminescence and fluorescence bacterial biosensors were closely comparable to each other. Our results demonstrate that both bacterial luminescence and fluorescence biosensors are useful in determining the presence and the bioavailable fractions of BTEX in the environment.     

Mutagenicity and genotoxicity assessment of industrial wastewaters

The genotoxicity of industrial wastewaters from Jajmau (Kanpur), was carried out by Ames Salmonella/microsome test, DNA repair-defective mutants, and Allium cepa anaphase–telophase test. Test samples showed maximum response with TA98 strain with and without metabolic activation. Amberlite resins concentrated wastewater samples were found to be more mutagenic as compared to those of liquid–liquid extracts (hexane and dichloromethane extracts). The damage in the DNA repair defective mutants in the presence of Amberlite resins concentrated water samples were found to be higher to that of liquid–liquid-extracted water samples at the dose level of 20 μl/ml culture. Among all the mutants, polA exhibited maximum decline with test samples. Mitotic index (MI) of root tip meristematic cells of A. cepa treated with 5, 10, 25, 50, and 100 % (v/v) wastewaters were significantly lower than the control. Complementary to the lower levels of MI, the wastewaters showed higher chromosomal aberration levels in all cases investigated.     

A simplified headspace biochemical oxygen demand test protocol based on oxygen measurements using a fiber optic probe.

Batch respirometric tests have many advantages over the conventional biochemical oxygen demand (BOD) method for analysis of wastewaters, including the use of nondiluted samples, a more rapid exertion of oxygen demand, and reduced sample preparation time. The headspace biochemical oxygen demand (HBOD) test can be used to obtain oxygen demands in 2 or 3 days that can predict 5-day biochemical oxygen demand (BOD5) results. The main disadvantage of the HBOD and other respirometric tests has been the lack of a simple and direct method to measure oxygen concentrations in the gas phase. The recent commercial production of a new type of fiber optic oxygen probe, however, provides a method to eliminate this disadvantage. This fiber optic probe, referred to here as the HBOD probe, was tested to see if it could be used in HBOD tests. Gas-phase oxygen measurements made with the HBOD probe took only a few seconds and were not significantly different from those made using a gas chromatograph (t test: n = 15, R2 = 0.9995, p < 0.001). In field tests using the HBOD probe procedure, the probe greatly reduced sample analysis time compared with previous HBOD and BOD protocols and produced more precise results than the BOD test for wastewater samples from two treatment plants (University Area Joint Authority [UAJA] Wastewater Treatment Plant in University Park, Pennsylvania, and The Pennsylvania State University [PSU] Wastewater Treatment Plant in University Park). Headspace biochemical oxygen demand measurements on UAJA primary clarifier effluent were 59.9 +/- 2.4% after 2 days (HBOD2) and 73.0 +/- 3.1% after 3 days (HBOD) of BOD, values, indicating that BOD5 values could be predicted by multiplying HBOD2 values by 1.67 +/- 0.07 or HBOD3 by 1.37 +/- 0.06. Similarly, tests using PSU wastewater samples could be used to provide BOD5 estimates by multiplying the HBOD2 by 1.24 +/- 0.04 or by multiplying the HBOD3 by 0.97 +/- 0.03. These results indicate that the HBOD fiber optic probe can be used to obtain reliable oxygen demands in batch respirometric tests such as the HBOD test.     

Airborne desert dust and aeromicrobiology over the Turkish Mediterranean coastline

Between 18 March and 27 October 2002, 220 air samples were collected on 209 of 224 calendar days, on top of a coastal atmospheric research tower in Erdemli, Turkey. The volume of air filtered for each sample was 340 liters. Two hundred fifty-seven bacterial and 2598 fungal colony forming units (CFU) were enumerated from the samples using a low-nutrient agar. Ground-based dust measurements demonstrated that the region is routinely impacted by dust generated regionally and from North Africa and that the highest combined percent recovery of total CFU and African dust deposition occurred in the month of April (93.4% of CFU recovery and 91.1% of dust deposition occurred during African dust days versus no African dust present, for that month). A statistically significant correlation was observed (peak regional African dust months of March, April and May; r_s=0.576, P=0.000) between an increase in the prevalence of microorganisms recovered from atmospheric samples on dust days (regional and African as determined by ground-based dust measurements), versus that observed on non-dust days. Given the prevalence of atmospherically suspended desert dust and microorganisms observed in this study, and that culture-based studies typically only recover a small fraction (<1.0%) of the actual microbial population in any given environment, dust-borne microorganisms and other associated constituents (organic detritus, toxins, etc.) may play a significant role in the regional human and ecosystem health.     

Combination of in vitro bioassays for the determination of cytotoxic and genotoxic potential of wastewater, surface water and drinking water samples

In this study we evaluated genotoxicity and cytotoxicity of native samples of wastewaters (15 samples), surface waters (28 samples) and potable waters (8 samples) with the SOS/umuC assay with Salmonella typhimurium TA1535/pSK1002 and MTT assay with human hepatoma HepG2 cells. The genotoxicity of selected samples was confirmed with the comet assay with HepG2 cells. In the SOS/umuC assay 13 out of the 51 samples were genotoxic: two effluent samples from chemical industry; one sample of wastewater treatment plant effluent; two hospital wastewater samples; three river water samples and four lake water samples. Six samples were cytotoxic for HepG2 cells: both effluent samples of chemical industry, two wastewater treatment plant effluent samples, and two river water samples, however, only the chemical industry effluent samples were genotoxic and cytotoxic, indicating that different contaminants are responsible for genotoxic and toxic effects. Comparing genotoxicity of river and lake water samples with the chemical analytical data of the presence of the residues of pharmaceutical and personal care products (non-steroidal anti-inflammatory drugs, UV filters and disinfectants) in these samples, indicated that the presence of UV filters might be linked to the genotoxicity of these samples. The results showed that the application of the bacterial SOS/umuC assay and mammalian cell assays (MTT and comet assay) with HepG2 cells was suitably sensitive combination of assays to monitor genotoxicity and cytotoxicity of native samples of wastewaters and surface waters. With this study we also confirmed that the toxicity/genotoxicity bioassays should be an integral tool in the evaluation of toxicity of complex wastewaters before the release into environment, as well as for the monitoring of surface water quality, providing data useful in risk assessment.     

MBBR/BAF工艺降解腈类有机物的性能研究

采用移动床生物膜反应器(MBBR)和曝气生物滤池(BAF)结合工艺对有机腈类废水处理进行了小试研究,以考察有机物的降解规律。经过稳定运行2个月,在两级硝化液回流比R=200%,pH=7.5,HRT=3 d的情况下,进水COD、CN-、NH3-N、TN及BOD5浓度平均值分别为3 024、38、185、305和845 mg/L时,出水COD、CN-、NH3-N、TN及BOD5平均浓度为50、0.5、10、45和17 mg/L,去除率分别为98.2%、98.7%、94.2%、85.2%和98.0%,BOD/COD比值由0.29提高到0.34,达到了《江苏省化学工业主要水污染物排放标准》(DB32/939-2006)一级排放标准。     

BOD5 measurements of water presenting inhibitory Cu2+. Implications in using of BOD to evaluate biodegradability of industrial wastewaters

In industrial effluents, the presence of an infinite number of possible mixtures of substances and the high variability of chemical conditions ask for an evaluation of biodegradability by a global and simple method. Biological oxygen demand after five days (BOD5) using synthetic wastewater was studied by two different ways: dilution and manometric methods. It can therefore be established that BOD5 obtained by adding manufactured inocula to the synthetic medium (effluent containing known and easily biodegradable substances) is close to the values obtained with inocula taken from the treated effluent of an urban and a rural purification plant. It was found that BOD5 measurement of effluents presenting factors affecting biodegradation, similar of those found in industrial effluents, is very questionable. The BOD is in this case influenced by the synergic and antagonist interactions between numerous and variable parameters like as pH, nature and concentration of inoculum, concentration of nutriments, amount and nature of assimilable substances, presence of toxicants, and presence of nitrification inhibitors, which are typical of real industrial wastewaters.     

Treatment of tannery wastewater in a pilot-scale hybrid constructed wetland system in Bangladesh

This paper reports the pollutant removal performances of a hybrid wetland system in Bangladesh for the treatment of a tannery wastewater. The system consisted of three treatment stages: a subsurface vertical flow (VF) wetland, followed by a horizontal flow (HF) and a VF wetland. The wetlands were planted with common reed (Phragmites australis), but employed different media, including organic coco-peat, cupola slag and pea gravel. In the first stage, experimental results demonstrated significant removal of ammonia (52%), nitrate (54%), BOD (78%), and COD (56%) under high organics loading rate (690 g COD m⁻² d⁻¹); simultaneous nitrification, denitrification, and organics degradation were attributed to the unique characteristics of the coco-peat media, which allowed greater atmospheric oxygen transfer for nitrification and organic degradation, and supply of organic carbon for denitrification. The second stage HF wetland produced an average PO₄ removal of 61%, primarily due to adsorption by the iron-rich cupola slag media. In the third treatment stage, which was filled with gravel media, further BOD removal (78%) from the tannery wastewater depleted organic carbon, causing the accumulation of NO₃ in the wastewater. Overall, the average percentage removals of NH₃-N, NO₃-N, BOD, COD, and PO₄ were 86%, 50%, 98%, 98% and 87%, respectively, across the whole hybrid system. The results provided a strong evidence to support widespread research and application of the constructed wetland as a low-cost, energy-efficient, wastewater treatment technology in Bangladesh.