The bioconversion of domestic wastewater sludge by immobilized mixed culture of filamentous fungi was investigated in a laboratory. The potential mixed culture of Penicillium corylophilum WWZA1003 and Aspergillus niger SCahmA103 was isolated from its local habitats (wastewater and sludge cake) and optimized on the basis of biodegradability and dewaterability of treated sludge. The observed results in this study showed that the sludge treatment was highly influenced by the effect of immobilized mixed fungi using liquid state bioconversion (LSB) process. The maximum production of dry filter cake (DFC) was enriched with fungal biomass to about 20.05 g/kg containing 23.47 g/kg of soluble protein after 4 days of fungal treatment. The reduction of COD, TSS, turbidity (optical density against distilled water, 660 nm), reducing sugar and protein in supernatant and filtration rate of treated sludge were influenced by the fungal mixed culture as compared to control (uninnoculated). After these processes, 99.4% of TSS, 98.05% of turbidity, 76.2% of soluble protein, 98% of reducing sugar and 92.4% of COD in supernatant of treated sludge were removed. Filtration time was decreased tremendously by the microbial treatment after 2 days of incubation. The effect of fungal strain on pH was also studied and presented. Effective bioconversion was observed after 4 days of fungal treatment.
Ten filamentous fungi adapted to domestic wastewater sludge (DWS) were further studied to evaluate their potential in terms of adaptation to higher sludge supplemented growing media and phytopathogenicity (induction of diseases to plants) to three germinating crop (Corn: Zea mays, Mung bean: Phaseolus aureus and Mustard: Brassica napus) seeds. The performances of the fungi in seed germination were evaluated based on percent germination index (GI) and infected/spotted seeds on direct fungal biomass (FBM) and fungal metabolite (FM). Significantly the highest biomass production was achieved with RW-P1 512 and Penicillium corylophilum (WW-P1003) at the highest (25%) sludge supplemented growing media that implied its excellent potentiality of adaptation and multiplication to domestic wastewater sludge. Significantly encouraging results of percent GI and spotted/infected seedlings were observed in FM than FBM by all fungi except the strain Aspergillus niger. A. niger gave the poorest percent of GI (24.30, 26.98 and 00.00%) and the highest percent of infected/spotted seeds (70, 100, and 100%) using FBM for corn, mung bean and mustard, respectively. On the other hand, comparatively the highest percent of GI (107.99, 106.25 and 117.67%) and the lowest percent of spotted/infected seedlings (3.3, 3.3 and 3.3%) were achieved with the isolate RW-P1 512 using FM. In FBM, the superior results of percent GI (86.61, 95.92 and 83.87%) and spotted/infected seedlings (3.3, 63.3 and 43.3%) were obtained by A. versicolor. Several crop seeds were responded differently for different fungal treatments. Hundred percent infected/spotted seeds in FM were recorded only for mustard with Trichoderma family that implied its strong sensitiveness to its metabolites.
Bioconversion of higher strength of domestic wastewater biosolids (sludge) (4% w/w of TSS) by mixed fungal culture of Aspergillus niger and Penicillium corylophilum was studied in a laboratory. The effect of potential mixed fungi on domestic wastewater sludge accelerated the liquid state bioconversion (LSB) process. The highest production of dry sludge cake (biosolids) was enriched with fungal biomass to about 85.66 g/kg containing 25.23 g/kg of protein after 8 days of treatment. The results presented in this study revealed that the reduction of chemical oxygen demand (COD), total suspended solid (TSS), and specific resistance to filtration (SRF) of treated sludge were highly influenced by the fungal culture as compared to control (uninnoculated). The maximum removal rates in treated sludge (biosolids) supernatant recorded were 92% of COD and 98.8% of TSS. Lower SRF (1.08 x 10(12) m/kg) was perceived in microbially treated sludge after 6 days of fermentation. The observed parameters were highly influenced after 8 days of treatment. The influence of pH was also studied and presented in the paper.
Effects of agitation and aeration rate on microbial treatment of domestic wastewater sludge were investigated in a batch fermenter using mixed culture of Penicillium corylophilum and Aspergillus niger. It was found that liquid state bioconversion (LSB) of wastewater sludge was highly influenced by the effects of agitation and aeration. The maximum production of sludge cake and reduction of organic substances in treated sludge were recorded at 150-200 rpm of agitation speed and 0.5 vvm of aeration rate after 72 h of treatment. No effective results were observed at higher rate of agitation (300 rpm) and aeration (1.5 vvm) as compared to optimum values. The results showed that the minimum level of air saturation (pO2) was adequate to maintain the bioconversion process.
A study was conducted to evaluate the settleability and dewaterability of fungal treated and untreated sludge using liquid state bioconversion process. The fungal mixed culture of Aspergillus niger and Penicillium corylophilum was used for fungal pretreatment of wastewater sludge. The fungal strains immobilized/entrapped on sludge particles with the formation of pellets and enhanced the separation process. The results presented in this study showed that the sludge particles (pellets) size of 2-5mm of diameter were formed with the microbial treatment of sludge after 2 days of fermentation that contained maximum 33.7% of total particles with 3-3.5mm of diameter. The settling rate (measured as total suspended solids (TSS) concentration, 130 mg/l) was faster in treated sludge than untreated sludge (TSS concentration, 440 mg/l) after 1 min of settling time. In 1 min of settling operation, 86.45% of TSS was settled in treated sludge while 4.35% of TSS settled in raw sludge. Lower turbidity was observed in treated sludge as compared to untreated sludge. The results to specific resistance to filtration (SRF) revealed that the fungal inoculum had significant potentiality to reduce SRF by 99.8% and 98.7% for 1% w/w and 4% w/w of TSS sludge, respectively. The optimum fermentation period recorded was 3 days for 1% w/w sludge and 6 days for 4% w/w sludge, respectively, for dewaterability test.
The biosolids accumulation and biodegradation of domestic wastewater treatment plant (DWTP) sludge by filamentous fungi have been investigated in a batch fermenter. The filamentous fungi Aspergillus niger and Penicillium corylophilum isolated from wastewater and DWTP sludge was used to evaluate the treatment performance. The optimized mixed inoculum (A. niger and P. corylophilum) and developed process conditions (co-substrate and its concentration, temperature, initial pH, inoculum size, and aeration and agitation rate) were incorporated to accelerate the DWTP sludge treatment process. The results showed that microbial treatment of higher strength of DWTP sludge (4% w/w of TSS) was highly influenced by the liquid state bioconversion (LSB) process. In developed bioconversion processes, 93.8 g/kg of biosolids was enriched with fungal biomass protein of 30 g/kg. Enrichment of nutrients such as nitrogen (N), phosphorous (P), potassium (K) in biosolids was recorded in 6.2% (w/w), 3.1% (w/w) and 0.15% (w/w) from its initial values of 4.8% (w/w), 2.0% (w/w) and 0.08% (w/w) respectively after 10 days of fungal treatment. The biodegradation results revealed that 98.8% of TSS, 98.2% of TDS, 97.3% of turbidity, 80.2% of soluble protein, 98.8% of reducing sugar and 92.7% of COD in treated DWTP sludge supernatant were removed after 8 days of microbial treatment. The specific resistance to filtration (SRF) in treated sludge (1.4x10(12) m/kg) was decreased tremendously by the microbial treatment of DWTP sludge after 6 days of fermentation compared to untreated sample (85x10(12) m/kg).
A study on liquid state bioconversion of sewage treatment plant (STP) sludge was assisted to evaluate the performance of batch fermenter compared to shake flask in a laboratory. Bioconversion of STP sludge was highly influenced by the mixed fungal culture of Penicillium corylophilum and Aspergillus niger after 4 days of treatment. The results showed that about 24.9 g kg(-1) dry sludge cake (DSC) was produced with enrichment of fungal biomass protein in fermenter while 20.1 g kg(-1) in shake flask after 4 days of fungal treatment. The effective biodegradation of STP sludge was recorded in both fermenter and shake flask experiment compared to control (uninnoculated sample). The results presented in this study revealed that the overall performance of fermenter in terms of sludge cake (biosolids) accumulation and biodegradation of STP sludge was higher than the shake flask.
A laboratory-scale study was undertaken to evaluate the liquid state bioconversion (LSB) in terms of biodegradation of microbially treated domestic wastewater sludge (biosolids) as well as its kinetics. The potential fungal strains and process factors developed from previous studies were used throughout the study. The results presented in this study showed that an effective biodegradation occurred with the biosolids (sludge cake) accumulated. The maximum biosolids (sludge cake) accumulated (93.8 g/kg of liquid sludge) enriched with the biomass protein (30.2 g/kg of dry biosolids), was achieved which improved the effluent quality by enhancing the removal of chemical oxygen demand (COD), reducing sugar (RS), soluble protein (SP), total dissolved solids (TDS), and total suspended solids (TSS). The higher reduction of specific resistance to filtration (SRF) was observed during bioconversion process. The kinetics results showed that the experimental data were better fitted for the biodegradation efficiency, and biosolids accumulation and biodegradation rate.
This study was undertaken to screen the filamentous fungi isolated from its relevant habitats(wastewater, sewage sludge and sludge cake) for the bioconversion of domestic wastewater sludge. A total of 35 fungal strains were tested against wastewater sludge (total suspended solids, TSS 1%-5% w/w) to evaluate its potentiality for enhancing the biodegradability and dewaterability using liquid state bioconversion(LSB) process. The strains were divided into five groups i.e. Penicillium, Aspergillus, Trichoderma, Basidiomycete and Miscellaneous, respectively. The strains WWZP1003, SCahmA103, SCahmT105 and PC-9 among their respective groups of Penicillium, Aspergillus, Trichoderma and Basidiomycete played potential roles in terms of separation (formation of pellets/flocs/filaments), biodegradation(removal of COD) and filtration (filterability) of treated domestic wastewater sludge. The Miscellaneous group was not considered due to its unsatisfactory results as compared to the other groups. The pH value was also influenced by the microbial treatment during fermentation process. The filterability of treated sludge was improved by fungal treatment, and lowest filtration time was recorded for the strain WWZP1003 and SCahmA103 of Penicillium and Aspergillus groups respectively compared with other strains.
The present study was designed to evaluate the potential of microbial adaptation and its affinity to biodegradation as well as bioconversion of soluble/insoluble (organic) substances of domestic wastewater treatment plant (DWTP) sludge (activated domestic sludge) under natural/non-sterilized conditions. The two filamentous fungi, Penicillium corylophilum (WWZP1003) and Aspergillus niger (SCahmA103) were used to achieve the objectives. It was observed that P. corylophilum (WWZP1003) was the better strain compared to A. niger (SCahmA103) for the bioconversion of domestic activated sludge through adaptation. The visual observation in plate culture showed that about 95-98% of cultured microbes (P. corylophilum and A. niger) dominated in treated sludge after 2 days of treatment. In this study, it was also found that the P. corylophilum was capable of removing 94.40% of COD and 98.95% of turbidity of filtrate with minimum dose of inoculum of 10% v/v in DWTP sludge (1% w/w). The pH level was lower (acidic condition) in the fungal treatment and maximum reduction of COD and turbidity was observed (at lower pH). The results for specific resistance to filtration (SRF) showed that the fungi played a great role in enhancing the dewaterability and filterability. In particular, the strain Penicillium had a more significant capability (than A. niger) of reducing 93.20% of SRF compared to the uninoculated sample. Effective results were observed by using fungal inoculum after 2 days of treatment. The developed LSB process is a new biotechnological approach for sludge management strategy.
A laboratory-scale study of bioconversion of local lignocellulosic material, oil palm biomass (OPB) was conducted by evaluating the enzyme production through microbial treatment in solid state bioconversion (SSB). OPB in the form of empty fruit bunches (EFB) was used as a solid substrate and treated with the white-rot fungus, Phanerochaete chrysosporium, to produce ligninase. The results showed that the highest ligninase activity of 400.27 U/liter was obtained at day 12 of fermentation. While the optimum study indicated the enzyme production of 1472.8 U/liter with moisture content of 50%, 578.7 U/liter with 10% v/w of inoculum size, and 721.8 U/liter with co-substrate concentration of 1% (w/w) at days 9, 9 and 12 of fungal treatment, respectively. The parameters glucosamine and reducing sugar were observed to evaluate the growth and substrate utilization in the experiment.
The study was attempted to produce activated carbons from palm oil mill effluent (POME) sludge. The adsorption capacity of the activated carbons produced was evaluated in aqueous solution of phenol. Two types of activation were followed, namely, thermal activation at 300, 500 and 800 degrees C, and physical activation at 15 degrees C (boiling treatment). A control (raw POME sludge) was used to compare the adsorption capacity of the activated carbons produced. The results indicated that the activation temperature of 800 degrees C showed maximum absorption capacity by the activated carbon (POME 800) in aqueous solution of phenol. Batch adsorption studies showed an equilibrium time of 6 h for the activated carbon of POME 800. It was observed that the adsorption capacity was higher at lower values of pH (2-3) and higher value of initial concentration of phenol (200-300 mg/L). The equilibrium data were fitted by the Langmuir and Freundlich adsorption isotherms. The adsorption of phenol onto the activated carbon POME 800 was studied in terms of pseudo- first and second order kinetics to predict the rate constant and equilibrium capacity with the effect of initial phenol concentrations. The rate of adsorption was found to be better correlation for the pseudo-second order kinetics compared to the first order kinetics.
Activated carbons derived from oil palm empty fruit bunches (EFB) were investigated to find the suitability of its application for removal of phenol in aqueous solution through adsorption process. Two types of activation namely; thermal activation at 300, 500 and 800 degrees C and physical activation at 150 degrees C (boiling treatment) were used for the production of the activated carbons. A control (untreated EFB) was used to compare the adsorption capacity of the activated carbons produced from these processes. The results indicated that the activated carbon derived at the temperature of 800 degrees C showed maximum absorption capacity in the aqueous solution of phenol. Batch adsorption studies showed an equilibrium time of 6 h for the activated carbon at 800 degrees C. It was observed that the adsorption capacity was higher at lower values of pH (2-3) and higher value of initial concentration of phenol (200-300 mg/L). The equilibrium data fitted better with the Freundlich adsorption isotherm compared to the Langmuir. Kinetic studies of phenol adsorption onto activated carbons were also studied to evaluate the adsorption rate. The estimated cost for production of activated carbon from EFB was shown in lower price (USD 0.50/kg of activated carbon) compared the activated carbon from other sources and processes.
The optimization of process parameters for the bioconversion of activated sludge by Penicillium corylophilum was investigated using response surface methodology (RSM). The three parameters namely temperature of 33 degrees C, agitation of 150 r/min, and pH of 5 were chosen as center point from the previous study of fungal treatment. The experimental data on chemical oxygen demand (COD) removal (%) were fitted into a quadratic polynomial model using multiple regression analysis. The optimum process conditions were determined by analyzing response surface three-dimensional surface plot and contour plot and by solving the regression model equation with Design Expert software. Box-Behnken design technique under RSM was used to optimize their interactions, which showed that an incubation temperature of 32.5 degrees C, agitation of 105 r/min, and pH of 5.5 were the best conditions. Under these conditions, the maximum predicted yield of COD removal was 98.43%. These optimum conditions were used to evaluate the trail experiment, and the maximum yield of COD removal was recorded as 98.5%.
A two-level fractional factorial design (FFD) was used to determine the effects of six factors, i.e. substrate (domestic wastewater sludge - DWS) and co-substrate concentration (wheat flour - WF), temperature, initial pH, inoculum size and agitation rate on the production of cellulase enzyme by Trichoderma harzianum in liquid state bioconversion. On statistical analysis of the results from the experimental studies, optimum process conditions were found to be temperature 32.5 degrees C, substrate concentration (DWS) 0.75% (w/w), co-substrate (WF) concentration 2% (w/w), initial pH 5, inoculum size 2% (v/w) and agitation 175 rpm. Analysis of variance (ANOVA) showed a high coefficient of determination (R2) of 0.975. Cellulase activity reached 10.2 FPU/ml at day 3 during the fermentation process which indicated about 1.5-fold increase in production compared to the cellulase activity obtained from the results of design of experiment (6.9 FPU/ml). Biodegradation of DWS was also evaluated to verify the efficiency of the bioconversion process as a waste management method.
Optimization of decolorization of methylene blue (MB) dye by lignin peroxidase (LiP) enzyme produced by white-rot fungus Phanerochaete chrysosporium using sewage treatment plant (STP) sludge as a major substrate was carried out in the laboratory. Optimization by the one-factor-at-a-time (OFAT) and statistical approach was carried out to determine the process conditions on optimum decolorization of MB dye using LiP enzyme in static mode. The OFAT method indicated that the optimum conditions for decolorization of MB dye (removal: 14-40%) was at temperature 55 degrees C, pH 5.0 with hydrogen peroxide (H(2)O(2)) concentration 4.0mM, MB dye concentration 20mg/L and LiP activity 0.487U/ml. The addition of veratryl alcohol to the reaction mixtures did not contribute any further increases in decolorization. The initial concentration of MB and the activity of LiP enzyme were further optimized using response surface methodology (RSM). The contour and surface plots suggested that the optimum initial concentration of MB and LiP activity predicted were 15mg/L and 0.687U/ml, respectively for the removal of 65%. The validation of the model showed that the decolorization process gave the higher removal of 90% in agitation mode compared to the static mode with 65% for 60min of incubation time by LiP enzyme.
A laboratory-scale study was carried out to produce lignin peroxidase (ligninase) by white rot fungus (Phanerochaete chrysosporium) using sewage-treatment-plant (STP) sludge as the major substrate. The optimization was done using full-factorial design (FFD) with agitation and aeration as the two parameters. Nine experiments indicated by the FFD were fermented in a stirred-tank bioreactor for 3 days. A second-order quadratic model was developed using the regression analysis of the experimental results with the linear, quadratic, and interaction effects of the parameters. Analysis of variance (ANOVA) showed a high coefficient of determination (R (2)) value of 0.972, thus indicating a satisfactory fit of the quadratic model with the experimental data. Using statistical analysis, the optimum aeration and agitation rates were determined to be 2.0 vvm and 200 rpm, respectively, with a maximum activity of 225 U l(-1) in the first 3 days of fermentation. The validation experiment showed the maximum activity of lignin peroxidase was 744 U l(-1) after 5 days of fermentation. The results for the tests of the stability of lignin peroxidase showed that the activity was more than 80% of the maximum for the first 12 h of incubation at an optimum pH of 5 and temperature of 55 degrees C.
A sequential optimization based on statistical design and one-factor-at-a-time (OFAT) method was employed to optimize the media constituents for the improvement of citric acid production from oil palm empty fruit bunches (EFB) through solid state bioconversion using Aspergillus niger IBO-103MNB. The results obtained from the Plackett-Burman design indicated that the co-substrate (sucrose), stimulator (methanol) and minerals (Zn, Cu, Mn and Mg) were found to be the major factors for further optimization. Based on the OFAT method, the selected medium constituents and inoculum concentration were optimized by the central composite design (CCD) under the response surface methodology (RSM). The statistical analysis showed that the optimum media containing 6.4% (w/w) of sucrose, 9% (v/w) of minerals and 15.5% (v/w) of inoculum gave the maximum production of citric acid (337.94 g/kg of dry EFB). The analysis showed that sucrose (p<0.0011) and mineral solution (p<0.0061) were more significant compared to inoculum concentration (p<0.0127) for the citric acid production.
The purpose of this study was to evaluate the feasibility of producing bioethanol from palm-oil mill effluent generated by the oil-palm industries through direct bioconversion process. The bioethanol production was carried out through the treatment of compatible mixed cultures such as Thrichoderma harzianum, Phanerochaete chrysosporium, Mucor hiemalis, and yeast, Saccharomyces cerevisiae. Simultaneous inoculation of T. harzianum and S. cerevisiae was found to be the mixed culture that yielded the highest ethanol production (4% v/v or 31.6 g/l). Statistical optimization was carried out to determine the operating conditions of the stirred-tank bioreactor for maximum bioethanol production by a two-level fractional factorial design with a single central point. The factors involved were oxygen saturation level (pO(2)%), temperature, and pH. A polynomial regression model was developed using the experimental data including the linear, quadratic, and interaction effects. Statistical analysis showed that the maximum ethanol production of 4.6% (v/v) or 36.3 g/l was achieved at a temperature of 32 degrees C, pH of 6, and pO(2) of 30%. The results of the model validation test under the developed optimum process conditions indicated that the maximum production was increased from 4.6% (v/v) to 6.5% (v/v) or 51.3 g/l with 89.1% chemical-oxygen-demand removal.
In this study, biodiesel was produced from sludge palm oil (SPO) using tolune-4-sulfonic monohydrate acid (PTSA) as an acid catalyst in different dosages in the presence of methanol to convert free fatty acid (FFA) to fatty acid methyl ester (FAME), followed by a transesterification process using an alkaline catalyst. In the first step, acid catalyzed esterification reduced the high FFA content of SPO to less than 2% with the different dosages of PTSA. The optimum conditions for pretreatment process by esterification were 0.75% (w/w) dosage of PTSA to SPO, 10:1 M ratio, 60 °C temperature, 60 min reaction time and 400 rpm stirrer speed. The highest yield of biodiesel after transesterification and purification processes was 76.62% with 0.07% FFA and 96% ester content. The biodiesel produced was favorable as compared to EN 14214 and ASTM 6751 standard. This study shows a potential exploitation of SPO as a new feedstock for the production of biodiesel.