Biomonitoring of wastewaters in treatment plants using ciliates

The aim of this project has been to study and compare the ciliate populations present in roptating biological reactors treating three different wastewaters. Wastewaters chosen were a maize mill (nejayote), a sugarcane/ethyl alcohol plant (vinasses) and a recycled paper mill (whitewaters). The initial dissolved organic contents, measured as soluble chemical oxygen demand (COD) and biochemical oxygen demand in five days (BOD₅), were 2040±150 mg COD L^–1 and 585±5 mg BOD₅L^–1 for nejayote; 2000±20 mg COD L^–1 and 640±5 mg BOD₅ L^–1 for vinasses and 960±200 mg COD L^–1 and 120±10 mg BOD₅ L^–1 for whitewaters. Results obtained indicate that ciliated protozoa proliferated in the different chambers of each rotating biological reactor (RBR). Saprobity indices, as a quantitative evaluation parameter, indicate that there are no universal species of ciliates associated with specific BOD₅ concentrations. Therefore, the number of species of ciliates present in the effluent indicate qualitatively the efficiency of removal of pollution from the wastewaters during treatment in the rotating biological reactors.     

Novel engineering controls to increase leachate contaminant degradation by refuse: From lab test to in situ engineering application

Here we provide direct evidence through a series laboratory and field-scale experiments using different age refuse to treat landfill leachate that aged refuse exhibits increased leachate contaminants removal ability with refuse stabilization time addition. Ten-years aged refuse showed best contaminant removal in a laboratory-scale test, removing 70.0% (8340.0-2540.0 mg/L) chemical oxygen demand (COD) and 75.0% (910.0-215.0 mg/L) ammonium-N, as well as removing 61.5-67.0% COD and 50.4-58.1% ammonium-N with variable COD (9948.0-12286.0 mg/L) and NH₃-N (780.0-1184.0 mg/L) in a field-scale test, respectively. When the 10-years aged refuse was disinfected by 20% NaClO (wt%), COD, biochemical oxygen demand (BOD₅), total nitrogen (TN), and ammonium-N removal showed a dramatic decrease throughout operation time from 84.4-86.2% to 15.2-34.5%, 94.4-99.8% to 26.2-54.4%, 31.2-33.9% to 2.1-10.1%, and 88.5-90.1% to 1.5-14.5%, respectively, suggesting biodegradation is the dominant contaminant removal. Based on this finding, a 3-stages (8 years) age refuse bioreactor (ARB) was constructed to treat leachate and ARB efficiently reduced chemical oxygen demand (COD) from 5478.0-10842.0 mg/L to 261.0-1020 mg/L (87.8-96.2% removal), ammonium-N from 811.4-1582.0 mg/L to 8.5-43.3 mg/L (96.9-99.4%), respectively, in 18 months running. In summary, the present studies suggest that increased leachate contaminant biodegradation ability of aged refuse could be used directly to create an engineering approach to treat leachate with operational and economic advantages.     

Treatment of coal gasification wastewater by a two-continuous UASB system with step-feed for COD and phenols removal

A two-continuous mesophilic (37 ± 2 °C) UASB system with step-feed was investigated as an attractive optimization strategy for enhancing COD and total phenols removal of the system and improving aerobic biodegradability of real coal gasification wastewater. Through the step-feed period, the maximum removal efficiencies of COD and total phenols reached 55-60% and 58-63% respectively in the system, at an influent flow distribution ratio of 0.2 and influent COD concentration of 2500 mg/L; the corresponding efficiencies were at low levels of 45-50% and 43-50% respectively at total HRT of 48 h during the single-feed period. The maximum specific methanogenic activity and substrate utilization rate were 592 ± 16 mg COD-CH₄/(gVSS d) and 89 ± 12 mg phenol/(gVSS d) during the step-feed operation. After the anaerobic digestion with step-feed, the aerobic effluent COD concentration decreased from 270 ± 9 to 215 ± 10 mg/L. The results suggested that step-feed enhanced the degradation of refractory organics in the second reactor.     

Thermophilic anaerobic digestion of Lurgi coal gasification wastewater in a UASB reactor

Lurgi coal gasification wastewater (LCGW) is a refractory wastewater, whose anaerobic treatment has been a severe problem due to its toxicity and poor biodegradability. Using a mesophilic (35 ± 2 °C) reactor as a control, thermophilic anaerobic digestion (55 ± 2 °C) of LCGW was investigated in a UASB reactor. After 120 days of operation, the removal of COD and total phenols by the thermophilic reactor could reach 50-55% and 50-60% respectively, at an organic loading rate of 2.5 kg COD/(m³ d) and HRT of 24 h; the corresponding efficiencies were both only 20-30% in the mesophilic reactor. After thermophilic digestion, the wastewater concentrations of the aerobic effluent COD could reach below 200 mg/L compared with around 294 mg/L if mesophilic digestion was done and around 375 mg/L if sole aerobic pretreatment was done. The results suggested that thermophilic anaerobic digestion improved significantly both anaerobic and aerobic biodegradation of LCGW.     

Removal of nitrate and phosphorus from hydroponic wastewater using a hybrid denitrification filter (HDF)

A laboratory-scale hybrid-denitrification filter (HDF) was designed by combining a plant material digester and a denitrification filter into a single unit for the removal of nitrate and phosphorus from glasshouse hydroponic wastewater. The carbon to nitrate (C:N) ratio for efficient operation of the HDF was calculated to be 1.93:1 and the COD/BOD₅ ratio was 1.2:1. When the HDF was continuously operated with the plant material replaced every 2 days and 100% internal recirculation of the effluent, a high level of nitrate removal (320–5 mg N/L, >95% removal) combined with a low effluent sBOD₅ concentration (<5 mg/L) was consistently achieved. Moreover, phosphate concentrations in the effluent were maintained below 7.5 mg P/L (>81% reduction). This study demonstrates the potential to combine a digester and a denitrification filter in a single unit to efficiently remove nitrate and phosphate from hydroponic wastewater in a single unit.     

Anaerobic digestion of whole stillage from dry-grind corn ethanol plant under mesophilic and thermophilic conditions

Anaerobic digestion of whole stillage from a dry-grind corn-based ethanol plant was evaluated by batch and continuous-flow digesters under thermophilic and mesophilic conditions. At whole corn stillage concentrations of 6348 to 50,786 mg total chemical oxygen demand (TCOD)/L, at standard temperature (0 °C) and pressure (1 atm), preliminary biochemical methane potential assays produced 88 ± 8 L (49 ± 5 L CH₄) and 96 ± 19 L (65 ± 14 L CH₄) biogas per L stillage from mesophilic and thermophilic digesters, respectively. Continuous-flow studies for the full-strength stillage (TCOD = 254 g/L) at organic loadings of 4.25, 6.30 and 9.05 g TCOD/L days indicated unstable performance for the thermophilic digester. Among the sludge retention times (SRTs) of 60, 45 and 30 days tested, the mesophilic digestion was successful only at 60 days-SRT which does not represent a practical operation time for a large scale bioethanol plant. Future laboratory studies will focus on different reactor configurations to reduce the SRT needed in the digesters.     

Textiles wastewater treatment using anoxic filter bed and biological wriggle bed-ozone biological aerated filter

In this study, the performance of the anoxic filter bed and biological wriggle bed-ozone biological aerated filter (AFB-BWB-O₃-BAF) process treating real textile dyeing wastewater was investigated. After more than 2 month process operation, the average effluent COD concentration of the AFB, BWB, O₃-BAF were 704.8 mg/L, 294.6 mg/L and 128.8 mg/L, with HRT being 8.1-7.7 h, 9.2 h and 5.45 h, respectively. Results showed that the effluent COD concentration of the AFB decreased with new carriers added and the average removal COD efficiency was 20.2%. During operation conditions, HRT of the BWB and O₃-BAF was increased, resulting in a decrease in the effluent COD concentration. However, on increasing the HRT, the COD reduction capability expressed by the unit carrier COD removal loading of the BWB reactor increased, while that of the O₃-BAF reactor decreased. This study is a beneficial attempt to utilize the AFB-BWB-O₃-BAF combine process for textile wastewater treatment.     

Effect of physical pretreatment on dilute acid hydrolysis of water hyacinth (Eichhornia crassipes)

Effects of different physical pretreatments on water hyacinth for dilute acid hydrolysis process (121 ± 3 °C, 5% H₂SO₄, 60 min) were comparatively investigated. Untreated sample had produced 24.69 mg sugar/g dry matter. Steaming (121 ± 3 °C) and boiling (100 ± 3 °C) for 30 min had provided 35.9% and 52.4% higher sugar yield than untreated sample, respectively. The highest sugar yield (132.96 mg sugar/g dry matter) in ultrasonication was obtained at 20 min irradiation using 100% power. The highest sugar production (155.13 mg sugar/g dry matter) was obtained from pulverized samples. Hydrolysis time was reduced when using samples pretreated by drying, mechanical comminution and ultrasonication. In most methods, prolonging the pretreatment period was ineffective and led to sugar degradations. Morphology inspection and thermal analysis had provided evidences of structure disruption that led to higher sugar recovery in hydrolysis process.     

Partial nitrification and nutrient removal in intermittently aerated sequencing batch reactors treating separated digestate liquid after anaerobic digestion of pig manure

The performance of an intermittently aerated sequencing batch reactor (IASBR) technology was investigated in achieving partial nitrification, organic matter removal and nitrogen removal from separated digestate liquid after anaerobic digestion of pig manure. The wastewater had chemical oxygen demand (COD) concentrations of 11,540 ± 860 mg/L, 5-day biochemical oxygen demand (BOD₅) concentrations of 2,900 ± 200 mg/L and total nitrogen (TN) concentrations of 4,041 ± 59 mg/L, with low COD:N ratios (2.9) and BOD₅:COD ratios (0.25). Synthetic wastewater, simulating the separated digestate liquid with similar COD and nitrogen concentrations but BOD₅ of 11,500 ± 100 mg/L, was also treated using the IASBR technology. At a mean organic loading rate of 1.15 kg COD/(m³ d) and a nitrogen loading rate of 0.38 kg N/(m³ d), the COD removal efficiency was 89.8% in the IASBR (IASBR-1) treating digestate liquid and 99% in the IASBR (IASBR-2) treating synthetic wastewater. The IASBR-1 effluent COD was mainly due to inert organic matter and can be further reduced to less than 40 mg/L through coagulation. The partial nitrification efficiency of 71–79% was achieved in the two IASBRs and one cause for the stable long-term partial nitrification was the intermittent aeration strategy. Nitrogen removal efficiencies were 76.5 and 97% in IASBR-1 and IASBR-2, respectively. The high nitrogen removal efficiencies show that the IASBR technology is a promising technology for nitrogen removal from low COD:N ratio wastewaters. The nitrogen balance analysis shows that 59.4 and 74.3% of nitrogen removed was via heterotrophic denitrification in the non-aeration periods in IASBR-1 and IASBR-2, respectively.     

Heat tolerance, behavioural temperature selection and temperature-dependent respiration in larval Octopus huttoni

This study reports temperature effects on paralarvae from a benthic octopus species, Octopus huttoni, found throughout New Zealand and temperate Australia. We quantified the thermal tolerance, thermal preference and temperature-dependent respiration rates in 1-5 days old paralarvae. Thermal stress (1 °C increase h⁻¹) and thermal selection (∼10-24 °C vertical gradient) experiments were conducted with paralarvae reared for 4 days at 16 °C. In addition, measurement of oxygen consumption at 10, 15, 20 and 25 °C was made for paralarvae aged 1, 4 and 5 days using microrespirometry. Onset of spasms, rigour (CT_max) and mortality (upper lethal limit) occurred for 50% of experimental animals at, respectively, 26.0±0.2 °C, 27.8±0.2 °C and 31.4±0.1 °C. The upper, 23.1±0.2 °C, and lower, 15.0±1.7 °C, temperatures actively avoided by paralarvae correspond with the temperature range over which normal behaviours were observed in the thermal stress experiments. Over the temperature range of 10 °C-25 °C, respiration rates, standardized for an individual larva, increased with age, from 54.0 to 165.2 nmol larvae⁻¹ h⁻¹ in one-day old larvae to 40.1-99.4 nmol h⁻¹ at five days. Older larvae showed a lesser response to increased temperature: the effect of increasing temperature from 20 to 25 °C (Q₁₀) on 5 days old larvae (Q₁₀=1.35) was lower when compared with the 1 day old larvae (Q₁₀=1.68). The lower Q₁₀ in older larvae may reflect age-related changes in metabolic processes or a greater scope of older larvae to respond to thermal stress such as by reducing activity. Collectively, our data indicate that temperatures >25 °C may be a critical temperature. Further studies on the population-level variation in thermal tolerance in this species are warranted to predict how continued increases in ocean temperature will limit O. huttoni at early larval stages across the range of this species.     

Contaminant removal efficiency depending on primary treatment and operational strategy in horizontal subsurface flow treatment wetlands

This study aimed to evaluate the contaminant removal efficiency of shallow horizontal subsurface flow treatment wetlands (SSF TWs) as a function of (1) primary treatment (hydrolytic upflow sludge blanket (HUSB) reactor vs. conventional settling) and (2) operation strategy (alternation of saturated/unsaturated phases vs. permanently saturated). An experimental plant was constructed, operated and surveyed for the main water quality parameters over a period of 2.5 years. The plant had 3 treatment lines: a control line (settler-wetland permanently saturated), a batch line (settler-wetland operated with saturated/unsaturated phases) and an anaerobic line (HUSB reactor-wetland permanently saturated). In each line wetlands had a surface area of 2.80 m², a water depth of 25 cm and a granular medium D₆₀ = 7.3 mm, and were planted with common reed. During the study period the wetlands were operated at a hydraulic and organic load of 28.5 mm/d and about 4.7 g BOD/m² d, respectively. Effluent average redox potential was lower for the anaerobic line (−45 ± 78 mV) than for the other two lines (3 ± 92.7 and −5 ± 71 mV for control and batch, respectively). Overall, chemical oxygen demand (COD), biochemical oxygen demand (BOD₅) and ammonium mass removal efficiencies were slightly greater for the batch line (88%, 96% and 87%, respectively) than for the control line (83%, 94% and 80%) and the anaerobic line (80%, 87% and 73%). During cold seasons, COD and ammonium removal in the batch line was around 30% and 50% higher than in the control line, respectively. The results of this study indicate that the implementation of a HUSB reactor as primary treatment did not enhance the treatment capacity of the system (in comparison with a conventional settler). The efficiency of treatment wetland systems with horizontal subsurface flow can be improved using a batch operation strategy.     

Effect of storage temperature on survival and infectivity of Steinernema rarum (OLI strain) (Rhabditida: Steinernematidae)

Nematode strains of the entomopathogenic family Steinernematidae differ in their ability to infect insects at different temperatures. Survival and infectivity of infective juveniles (IJs) of Steinernema rarum (OLI) were studied after their storage at 23 ± 2 °C and at 5 ± 1 °C. Survival at 23 ± 2 °C was always above 95%. At 5 ± 1 °C, survival decreased at week 5, but infectivity did the same after week 2. Unlike other steinernematids, both infectivity and survival of IJs would be higher for S. rarum (OLI) when stored at 23 ± 2 °C.     

Decolorization of indigo carmine by laccase displayed on Bacillus subtilis spores

Blue multicopper oxidases, laccases displayed on the surface of Bacillus spores were used to decolorize a widely used textile dyestuff, indigo carmine. The laccase-encoding gene of Bacillus subtilis, cotA, was cloned and expressed in B. subtilis DB104, and the expressed enzyme was spontaneously localized on Bacillus spores. B. subtilis spores expressing laccase exhibited maximal activity for the oxidation of 2,2′-azino-bis (3-ethylthiazoline-6-sulfonate) (ABTS) at pH 4.0 and 80 °C, and for the decolorization of indigo carmine at pH 8.0 and 60 °C. The displayed enzyme retained 80% of its original activity after pre-treatment with organic solvents such as 50% acetonitrile and n-hexane for 2 h at 37 °C. The apparent K_m of the enzyme displayed on spores was 443 ± 124 μM for ABTS with a V_max of 150 ± 16 U/mg spores. Notably, 1 mg of spores displaying B. subtilis laccase (3.4 × 10² U for ABTS as a substrate) decolorized 44.6 μg indigo carmine in 2 h. The spore reactor (0.5 g of spores corresponding to 1.7 × 10⁵ U in 50 mL) in a consecutive batch recycling mode decolorized 223 mg indigo carmine/L to completion within 42 h at pH 8.0 and 60 °C. These results suggest that laccase displayed on B. subtilis spores can serve as a powerful environmental tool for the treatment of textile dye effluent.     

The treatment of cheese whey wastewater by sequential anaerobic and aerobic steps in a single digester at pilot scale

The treatment of reconstituted whey wastewater was performed in a 400 L digester at 20 °C, with an anaerobic digestion step, followed by a step of aerobic treatment at low oxygen concentration in the same digester. In a first set of 48 cycles, total cycle time (T_C) of 2, 3 and 4 days were tested at varying organic loading rates (OLR). The COD removal reached 89 ± 4, 97 ± 3 and 98 ± 2% at T_C of 2, 3 and 4 days and OLR of 0.56, 1.04 and 0.78 gCOD L⁻¹ d⁻¹, respectively. The activity of the biomass decreased for the methanogenic population, while increasing by 400% for the acidogens, demonstrating a displacement in the predominant trophic group in the biomass bed. A second set of 16 cycles was performed with higher soluble oxygen concentration in the bulk liquid (0.5 mg L⁻¹) during the aerobic treatment at a T_C of 2 days and an OLR of 1.55 gCOD L⁻¹ d⁻¹, with a soluble COD removal of 88 ± 3%. The biomass specific activities showed a compartmentalization of the trophic group with methanogenic activity maintained in the biomass bed and a high acidogenic activity in the suspended flocs.     

A kinetic evaluation of anaerobic treatment of swine wastewater at two temperatures in a temperate climate zone

A static granular bed reactor (SGBR) was used to treat swine wastewater at 24 and 16 °C. At 24 °C, the organic loading rate (OLR) was 0.7-5.4 kg COD/m³ day and the average chemical oxygen demand (COD) removal efficiency was 88.5%, respectively. Meanwhile, at 16 °C, the OLR was 1.6-4.0 kg COD/m³ day and the average COD removal efficiency was 68.0%, respectively. The SGBR acted as a bioreactor as well as a biofilter. After backwashing, the recovery of COD removal was not a function of an OLR but recovery time, while that of TSS removal was not a function of either recovery time or the OLR. The maximum substrate utilization rate (k_max) ratio was 1.89 between 24 and 16 °C, and the half velocity constant (K_s) ratio was 1.22, and the maximum specific growth rate (μ_max) ratio was 4.71. In addition, the temperature-activity coefficient in this study was determined to be 1.09.     

Electricity generation in single-chamber microbial fuel cells using a carbon source sampled from anaerobic reactors utilizing grass silage

Production of electricity from samples obtained during anaerobic digestion of grass silage was examined using single-chamber air-cathode mediator-less microbial fuel cells (MFCs). The samples were obtained from anaerobic reactors at start-up conditions after 3 and 10 days of operation under psychrophilic (15 °C) and mesophilic (37 °C) temperatures. Electricity was directly produced from all samples at a concentration of 1500 mg COD L⁻¹. Power density obtained from the samples, as a sole carbon source, ranged from 56 ± 3 W m⁻³ to 31 ± 1 W m⁻³ for the mesophilic and psychrophilic samples, respectively. Coulombic efficiencies ranged from 18 ± 1% to 12 ± 1% for the same samples. The relationship between the maximum voltage output and initial COD concentration appeared to follow saturation kinetics at the external resistance of 217 Ω. Chemical oxygen demand (COD) removal was over 90% and total phenolics removal was in the range of 30-75% for all samples tested, with a standard amount of 60 mg L⁻¹ total phenolics removed for every sample. Our results indicate that generating electricity from solution samples of anaerobic reactors utilizing grass silage is possible, opening the possibility for combination of anaerobic digestion with MFC technology for energy generation.     

Sulfidogenic fluidized bed treatment of real acid mine drainage water

The treatment of real acid mine drainage water (pH 2.7-4.3) containing sulfate (1.5-3.34 g/L) and various metals was studied in an ethanol-fed sulfate-reducing fluidized bed reactor at 35 °C. The robustness of the process was tested by increasing stepwise sulfate, ethanol and metal loading rates and decreasing feed pH and hydraulic retention time. Highest sulfate reduction rate (4.6 g/L day) was obtained with feed sulfate concentration of 2.5 g/L, COD/sulfate ratio of 0.85 and HRT of 12 h. The corresponding sulfate and COD removal efficiencies were about 90% and 80%, respectively. The alkalinity produced in sulfidogenic ethanol oxidation neutralized the acidic mine water. Highest metal precipitation efficiencies were observed at HRT of 24 h, the percent metal removal being over 99.9% for Al (initial concentration 55 mg/L), Co (9.0 mg/L), Cu (49 mg/L), Fe (435 mg/L), Ni (3.8 mg/L), Pb (7.5 mg/L) and Zn (6.6 mg/L), and 94% for Mn (7.21 mg/L).     

Bio-kinetic analysis on treatment of textile dye wastewater using anaerobic batch reactor

An anaerobic digestion technique was applied to textile dye wastewater aiming at the colour and COD removal. Pet bottles of 5 L capacity were used as reactor which contains methanogenic sludge of half a liter capacity which was used for the treatment of combined synthetic textile dye and starch wastewater at different mixing ratios of 20:80, 30:70, 40:60, 50:50 and 60:40 with initial COD concentrations as 3520, 3440, 3360, 3264 and 3144 mg L⁻¹, respectively. The reactor was maintained at room temperature (30 ± 3 °C) with initial pH of 7. The maximum COD and colour removal were 81.0% and 87.3% at an optimum mixing ratio of 30:70 of textile dye and starch wastewaters. Both Monod’s and Haldane’s models were adopted in this study. The kinetic constants of cell growth under Haldane’s model were satisfactory when compared to Monod’s model. The kinetic constants obtained by Haldane’s model were found to be in the range of μ_max = 0.037-0.146 h⁻¹, K_s = 651.04-1372.88 mg L⁻¹ and K_i = 5681.81-18727.59 mg L⁻¹.     

Performance of sulfidogenic anaerobic baffled reactor (ABR) treating acidic and zinc-containing wastewater

The applicability of anaerobic baffled reactor (ABR) was investigated for the treatment of acidic (pH 4.5–7.0) wastewater containing sulfate (1000–2000 mg/L) and Zn (65–200 mg/L) at 35 °C. The ABR consisted of four equal stages and lactate was supplemented (COD/SO₄²⁻ = 0.67) as carbon and energy source for sulfate reducing bacteria (SRB). The robustness of the system was studied by decreasing pH and increasing Zn, COD, and sulfate loadings. Sulfate-reduction efficiency quickly increased during the startup period and reached 80% within 45 days. Decreasing feed pH, increasing feed sulfate and Zn concentrations did not adversely affect system performance as sulfate reduction and COD removal efficiencies were within 62–90% and 80–95%, respectively. Although feed pH was steadily decreased from 7.0 to 4.5, effluent pH was always within 6.8–7.5. Over 99% Zn removal was attained throughout the study due to formation of Zn-sulfide precipitate.