A crude oil spill is a common issue during offshore oil drilling, transport and transfer to onshore. Second, the production of petroleum refinery effluent is known to cause pollution due to its toxic effluent discharge. Sea habitats and onshore soil biota are affected by total petroleum hydrocarbons (TPH) as a pollutant in their natural environment. Crude oil pollution in seawater, estuaries and beaches requires an efficient process of cleaning. To remove crude oil pollutants from seawater, various physicochemical and biological treatment methods have been applied worldwide. A biological treatment method using bacteria, fungi and algae has recently gained a lot of attention due to its efficiency and lower cost. This review introduces various studies related to the bioremediation of crude oil, TPH and related petroleum products by bioaugmentation and biostimulation or both together. Bioremediation studies mentioned in this paper can be used for treatment such as emulsified residual spilled oil in seawater with floating oil spill containment booms as an enclosed basin such as a bioreactor, for petroleum hydrocarbons as a pollutant that will help environmental researchers solve these problems and completely clean-up oil spills in seawater.
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).
The potential use of n-dodecane and n-hexadecane as oxygen vectors for enhancing hyaluronic acid (HA) biosynthesis by Streptococcus zooepidemicus ATCC 39920 was investigated using a 2-L stirred-tank bioreactor equipped with helical ribbon or Rushton turbine impellers. The volumetric fraction of the oxygen vector influenced the gas-liquid volumetric oxygen transfer coefficient (K(L)a) positively. Batch HA fermentation with 1% (v/v) n-dodecane or 0.5% (v/v) n-hexadecane addition was carried out at different impeller tip speeds. Even though cell growth was lower in the fermentation with oxygen vector addition, the HA productivity and molecular weight were higher when compared to the fermentation without oxygen vector at low impeller tip speed. The highest HA concentration (4.25 gHA/l) and molecular weight (1.54 × 10(7) Da) were obtained when 0.5% (v/v) n-hexadecane and 0.785 m/s impeller tip speed of helical ribbon were used.
Two-stage fermentation was normally employed to achieve a high poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] productivity with higher 4HB molar fraction. Here, we demonstrated single-stage fermentation method which is more industrial feasible by implementing mixed-substrate cultivation strategy. Studies on bioreactor scale show a remarkably high PHA accumulation of 73 wt%, contributing to a high PHA concentration and product yield of 8.6 g/L and 2.7 g/g, respectively. This fermentation strategy has resulted in copolymers with wider range of 4HB monomer composition, which ranges from 12 to 55 mol%. These copolymers show a broad range of weight average molecular weight (M w ) from 119.5 to 407.0 kDa. The copolymer characteristics were found to be predominantly affected by the nature of the substrates and the mixture strategies, regardless of the 4HB monomer compositions. This was supported by the determination of copolymer randomness using (13)C-NMR analysis. The study warrants significantly in the copolymer scale-up and modeling at industrial level.
Two cocoa bean fermentation methods (spontaneous fermentation and the use of starter culture) for 7 days fermentation were compared in terms of safety and quality fermented beans. Candida sp. was used as a starter culture in this study. The safety of the fermented cocoa beans were measured by the growth colonies of pathogenic microorganisms namely Bacillus cereus, Escherichia coli, Salmonella sp., Staphylococcus aureus, and Pseudomonas sp., on Bacillus cereus agar, eosin-methylene blue (EMB) agar, xylose lysine deoxycholate (XLD) agar, Baird-Parker agar (BPA), and Pseudomonas agar, respectively. B. cereus, E. coli and Salmonella sp. were early present in both fermentations. Candida sp.-fermentation showed detection of B. cereus at 5.34 log10 CFU/g and absence after 24 hours of fermentation while in spontaneous-fermentation B. cereus was too few to count. Moreover, the log10 E. coli number in Candida sp.-fermentation and spontaneous-fermentation were reduced from 5.72 to 3.66 and from 7.15 to 4.46 on day 1 to day 3, respectively. There were no presences of pathogenic microorganisms on day 5 and day 7 for both fermentations. In term of quality, proximate analysis of spontaneous-fermentation resulted that the content of moisture, ash, fat, crude protein, crude fibre and carbohydrate was 56.47%, 2.32%, 3.17%, 7.02%, 28.14% and 2.88%, meanwhile for the Candida sp.-fermentation was 53.96%, 2.19%, 3.44%, 8.25%, 25.46% and 6.70%, respectively. This study showed that both fermentations are considered to be safe and there is no significant difference in proximate value in fermented cocoa beans from spontaneous-fermentation and Candida sp.-fermentation.
In Malaysia, there has been interest in the utilization of palm oil and oil palm biomass for the production of environmental friendly biofuels. A biorefinery based on palm oil and oil palm biomass for the production of biofuels has been proposed. The catalytic technology plays major role in the different processing stages in a biorefinery for the production of liquid as well as gaseous biofuels. There are number of challenges to find suitable catalytic technology to be used in a typical biorefinery. These challenges include (1) economic barriers, (2) catalysts that facilitate highly selective conversion of substrate to desired products and (3) the issues related to design, operation and control of catalytic reactor. Therefore, the catalytic technology is one of the critical factors that control the successful operation of biorefinery. There are number of catalytic processes in a biorefinery which convert the renewable feedstocks into the desired biofuels. These include biodiesel production from palm oil, catalytic cracking of palm oil for the production of biofuels, the production of hydrogen as well as syngas from biomass gasification, Fischer-Tropsch synthesis (FTS) for the conversion of syngas into liquid fuels and upgrading of liquid/gas fuels obtained from liquefaction/pyrolysis of biomass. The selection of catalysts for these processes is essential in determining the product distribution (olefins, paraffins and oxygenated products). The integration of catalytic technology with compatible separation processes is a key challenge for biorefinery operation from the economic point of view. This paper focuses on different types of catalysts and their role in the catalytic processes for the production of biofuels in a typical palm oil and oil palm biomass-based biorefinery.
Tricalcium phosphate ceramic microcarrier has been developed and introduced to a new possibility for the culture of anchorage dependent animal cells of DF1. It was observed that the number of attached cells was increased with shorter time for both spinner vessel and stirred tank (ST) bioreactor. For those bioreactors, the total viable cell number that had been obtained is about 1.2 x 10(5) cell/ml.
Microbial flocs formed from raw textile wastewater in a prototype Aerobic Biofilm Reactor (ABR) system were characterised and studied for their potential use in the treatment of textile wastewater. After 90-100 days of operation, microbial flocs of loose irregular structures were obtained from the reactor with good settling velocity of 33 m/h and sludge volume index (SVI) of 48.2 mL/g. Molecular analysis of the flocs using PCR-amplified 16S rDNA sequence showed 98% homology to those of Bacillus sp, Paenibacillus sp and Acromobacter sp. Detection of Ca(2+)(131 mg/g) and Fe(2+)(131 mg/g) using atomic absorption spectrometer might be implicated with the flocs formation. In addition, presence of Co(2+) and Ni(2+) were indicative of the flocs ability to accumulate at least a fraction of the metals' present in the wastewater. When the flocs were used for the treatment of raw textile wastewater, they showed good removal of COD and colour about 55% and 70% respectively, indicating their potential application.
The influence of hydraulic retention time (HRT, 24, 12, and 6h) on the physical characteristics of granules and performance of a sequencing batch reactor (SBR) treating rubber wastewater was investigated. Results showed larger granular sludge formation at HRT of 6h with a mean size of 2.0±0.1mm, sludge volume index of 20.1mLg(-1), settling velocity of 61mh(-1), density of 78.2gL(-1) and integrity coefficient of 9.54. Scanning electron microscope analyses revealed different morphology of microorganisms and structural features of granules when operated at various HRT. The results also demonstrated that up to 98.4% COD reduction was achieved when the reactor was operated at low HRT (6h). Around 92.7% and 89.5% removal efficiency was noted for ammonia and total nitrogen in the granular SBR system during the treatment of rubber wastewater.
Textile wastewater, one of the most polluted industrial effluents, generally contains substantial amount of dyes and chemicals that will cause increase in the COD, colour and toxicity of receiving water bodies if not properly treated. Current treatment methods include chemical and biological processes; the efficiency of the biological treatment method however, remains uncertain since the discharged effluent is still highly coloured. In this study, granules consisting mixed culture of decolourising bacteria were developed and the physical and morphological characteristics were determined. After the sixth week of development, the granules were 3-10 mm in diameter, having good settling property with settling velocity of 70 m/h, sludge volume index (SVI) of 90 to 130 mL/g, integrity coefficient of 3.7, and density of 66 g/l. Their abilities to treat sterilised raw textile wastewater were evaluated based on the removal efficiencies of COD (initial ranging from 200 to 3,000 mg/L), colour (initial ranging from 450 to 2000 ADMI) of sterilised raw textile wastewater with pH from 6.8 to 9.4. Using a sequential anaerobic-aerobic treatment cycle with hydraulic retention time (HRT) of 24 h, maximum removal of colour and COD achieved was 90% and 80%, respectively.
The concept of de novo metabolic engineering through novel synthetic pathways offers new directions for multi-step enzymatic synthesis of complex molecules. This has been complemented by recent progress in performing enzymatic reactions using immobilized enzyme microreactors (IEMR). This work is concerned with the construction of de novo designed enzyme pathways in a microreactor synthesizing chiral molecules. An interesting compound, commonly used as the building block in several pharmaceutical syntheses, is a single diastereoisomer of 2-amino-1,3,4-butanetriol (ABT). This chiral amino alcohol can be synthesized from simple achiral substrates using two enzymes, transketolase (TK) and transaminase (TAm). Here we describe the development of an IEMR using His6-tagged TK and TAm immobilized onto Ni-NTA agarose beads and packed into tubes to enable multi-step enzyme reactions. The kinetic parameters of both enzymes were first determined using single IEMRs evaluated by a kinetic model developed for packed bed reactors. The Km(app) for both enzymes appeared to be flow rate dependent, while the turnover number kcat was reduced 3 fold compared to solution-phase TK and TAm reactions. For the multi-step enzyme reaction, single IEMRs were cascaded in series, whereby the first enzyme, TK, catalyzed a model reaction of lithium-hydroxypyruvate (HPA) and glycolaldehyde (GA) to L-erythrulose (ERY), and the second unit of the IEMR with immobilized TAm converted ERY into ABT using (S)-α-methylbenzylamine (MBA) as amine donor. With initial 60mM (HPA and GA each) and 6mM (MBA) substrate concentration mixture, the coupled reaction reached approximately 83% conversion in 20 min at the lowest flow rate. The ability to synthesize a chiral pharmaceutical intermediate, ABT in relatively short time proves this IEMR system as a powerful tool for construction and evaluation of de novo pathways as well as for determination of enzyme kinetics.
Understanding the relationship between microbial community and mechanism of aerobic granulation could enable wider applications of granules for high-strength wastewater treatment. The majority of granulation studies principally determine the engineering aspects of granules formation with little emphasis on the microbial diversity. In this study, three identical reactors namely R1, R2 and R3 were operated using POME at volumetric loadings of 1.5, 2.5 and 3.5 kg COD m(-3) d(-1), respectively. Aeration was provided at a volumetric flow rate of 2.5 cms(-1). Aerobic granules were successfully developed in R2 and R3 while bioflocs dominated R1 until the end of experiments. Fractal dimension (D(f)) averaged at 1.90 suggesting good compactness of granules. The PCR-DGGE results indicated microbial evolutionary shift throughout granulation despite different operating OLRs based on decreased Raup and Crick similarity indices upon mature granule formation. The characteristics of aerobic granules treating high strength agro-based wastewater are determined at different volumetric loadings.
Thiocyanate (SCN-) forms as a by-product of cyanidation during gold ore processing and can be degraded by a variety of microorganisms utilizing it as an energy, nitrogen, sulphur and/or carbon source. In complex consortia inhabiting bioreactor systems, a range of metabolisms are sustained by SCN- degradation; however, despite the addition or presence of labile carbon sources in most bioreactor designs to date, autotrophic bacteria have been found to dominate key metabolic functions. In this study, we cultured an autotrophic SCN--degrading consortium directly from gold mine tailings. In a batch-mode bioreactor experiment, this consortium degraded 22 mM SCN-, accumulating ammonium (NH4+) and sulphate (SO42-) as the major end products. The consortium consisted of a diverse microbial community comprised of chemolithoautotrophic members, and despite the absence of an added organic carbon substrate, a significant population of heterotrophic bacteria. The role of eukaryotes in bioreactor systems is often poorly understood; however, we found their 18S rRNA genes to be most closely related to sequences from bacterivorous Amoebozoa. Through combined chemical and phylogenetic analyses, we were able to infer roles for key microbial consortium members during SCN- biodegradation. This study provides a basis for understanding the behaviour of a SCN- degrading bioreactor under autotrophic conditions, an anticipated approach to remediating SCN- at contemporary gold mines.
Nine aerobic cellulolytic bacterial cultures were obtained from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Culture (DSMZ) and the American Type Culture Collection (ATCC). The objectives of this study were to characterize the cellulolytic bacteria and to determine the optimum moisture ratio required for solid state fermentation (SSF) of palm kernel cake (PKC). The bacteria cultures were grown on reconstituted nutrient broth, incubated at 30°C and agitated at 200 rpm. Carboxymethyl cellulase, xylanase, and mannanase activities were determined using different substrates and after SSF of PKC. The SSF was conducted for 4 and 7 days with inoculum size of 10% (v/w) on different PKC concentration-to-moisture ratios: 1 : 0.2, 1 : 0.3, 1 : 0.4, and 1 : 0.5. Results showed that Bacillus amyloliquefaciens 1067 DSMZ, Bacillus megaterium 9885 ATCC, Paenibacillus curdlanolyticus 10248 DSMZ, and Paenibacillus polymyxa 842 ATCC produced higher enzyme activities as compared to other bacterial cultures grown on different substrates. The cultures mentioned above also produced higher enzyme activities when they were incubated under SSF using PKC as a substrate in different PKC-to-moisture ratios after 4 days of incubation, indicating that these cellulolytic bacteria can be used to degrade and improve the nutrient quality of PKC.
Many developing countries, mostly situated in the tropical region, have incorporated a biological nitrogen removal process into their wastewater treatment plants (WWTPs). Existing wastewater characteristic data suggested that the soluble chemical oxygen demand (COD) in tropical wastewater is not sufficient for denitrification. Warm wastewater temperature (30 °C) in the tropical region may accelerate the hydrolysis of particulate settleable solids (PSS) to provide slowly-biodegradable COD (sbCOD) for denitrification. This study aimed to characterize the different fractions of COD in several sources of low COD-to-nitrogen (COD/N) tropical wastewater. We characterized the wastewater samples from six WWTPs in Malaysia for 22 months. We determined the fractions of COD in the wastewater by nitrate uptake rate experiments. The PSS hydrolysis kinetic coefficients were determined at tropical temperature using an oxygen uptake rate experiment. The wastewater samples were low in readily-biodegradable COD (rbCOD), which made up 3-40% of total COD (TCOD). Most of the biodegradable organics were in the form of sbCOD (15-60% of TCOD), which was sufficient for complete denitrification. The PSS hydrolysis rate was two times higher than that at 20 °C. The high PSS hydrolysis rate may provide sufficient sbCOD to achieve effective biological nitrogen removal at WWTPs in the tropical region.
Sugarcane vinasse has been reported as a high strength industrial wastewater that could cause severe environmental pollution due to its complex and bio-refractory compounds. Thus, the combined coagulation and sequencing batch biofilm reactor (SBBR) system was employed for the sugarcane vinasse treatment. This study aims to determine the recommended conditions of various parameters under coagulation and SBBR and investigate the effectiveness of combined processes. First, the approach of the coagulation process could achieve the maximum COD reduction and decolorization efficiencies of 79.0 ± 3.4% and 94.1 ± 1.9%, respectively, under the recommended conditions. Next, SBBR as an integrated biofilm reactor showed excellent synergistic biodegradability, removing 86.6 ± 4.3% COD concentration and 94.6 ± 3.8% color concentration at 3.0 g·COD/L of substrate loading concentration. The kinetic studies of SBBR revealed that the first-order kinetic model was the best fit for COD reduction efficiency. In contrast, the second-order kinetic model was the best fit for decolorization efficiency. The SBBR reaction was further investigated by ultraviolet-visible spectrophotometry (UV-Vis). In the combined processes, SBBR followed by the coagulation process (SBBR-CP) showed greater COD reduction and decolorization efficiencies (97.5 ± 0.3 and 99.4 ± 0.1%) when compared to the coagulation process followed by SBBR (CP-SBBR). This study demonstrated the removal performance and potential application of the combined sequential process to produce effluent that can be reused for bioethanol production and fertigation. This finding provides additional insight for developing effective vinasse treatment using combined chemical and biological processes.
Lack of sufficient nitrogenous substrate and buffering potential have been acknowledged as impediments to the treatment of palm oil mill effluent through co-digestion processes. In this study, ammonium bicarbonate was used to provide the nitrogenous substrate and buffering potential. To regulate the impact of ammonium bicarbonate toxicity on the anaerobic co-digestion system, dosages from 0 to 40 mg/L were supplemented. The biogas yield was used to indicate the effects of NH4+ toxicity. In a solar-assisted bioreactor, solar radiation was first collected by a solar panel and converted into electricity, which was then used to heat a mixture of palm oil mill effluent and cattle manure to maintain the reactor in the mesophilic temperature range. This co-digestion operation was performed semi-continuously and was analyzed at a 50:50 mixing ratio of palm oil mill effluent and cattle manure. The results indicate that the additional dosing of ammonium bicarbonate can significantly enhance biogas production. Maximum cumulative biogas and methane productions of 2034.00 mL and 1430.51 mL, respectively, were obtained with the optimum addition of 10 mg/L ammonium bicarbonate; these values are 29.80% and 42.30% higher, respectively, than that obtained in the control co-digestion operation without addition of ammonium bicarbonate. Utilization of a mathematical equation (G = Gmk/t) to describe a kinetic analysis of the biogas yield also indicated that the optimum ammonium bicarbonate dose was 10 mg/L. The results of this study suggest that supplementation with ammonium bicarbonate doses of up to 40 mg/L can be used to provide nitrogenous substrates and buffering potential in anaerobic co-digestion processes. The determination of the optimal dose provides an alternative and efficient option for enhanced biogas production, which will have obvious economic advantages for feasible industrial applications.
The complexity of residual toxic organics from biologically treated effluents of pulp and paper mills is a serious concern. To date, it has been difficult to choose the best treatment technique because each of the available options has advantages and drawbacks. In this study, two different treatment techniques using laboratory-scale aerobic sequencing batch reactors (SBRs) were tested with the same real recycled paper mill effluent to evaluate their treatment efficiencies. Two attached-growth SBRs using granular activated carbon (GAC) with and without additional biomass and a suspended-growth SBR were used in the treatment of real recycled paper mill effluent at a chemical oxygen demand (COD) level in the range of 800-1300 mg/L, a fixed hydraulic retention time of 24 h and a COD:N:P ratio of approximately 100:5:1. The efficiency of this biological treatment process was studied over a 300-day period. The six most important wastewater quality parameters, namely, chemical oxygen demand (COD), turbidity, ammonia (expressed as NH3-N), phosphorus (expressed as PO4(3)-P), colour, and suspended solids (SS), were measured to compare the different treatment techniques. It was determined that these processes were able to almost completely and simultaneously eliminate COD (99%) and turbidity (99%); the removals of NH3-N (90-100%), PO4(3)-P (66-78%), colour (63-91%), and SS (97-99%) were also sufficient. The overall performance results confirmed that an attached-growth SBR system using additional biomass on GAC is a promising configuration for wastewater treatment in terms of performance efficiency and process stability under fluctuations of organic load. Hence, this hybrid system is recommended for the treatment of pulp and paper mill effluents.
A comprehensive exergoeconomic performance analysis of a municipal solid waste digestion plant integrated with a biogas genset was conducted throughout this study in order to highlight its bottlenecks for further improvements. Exergoeconomic performance parameters of each component of the plant were determined by solving exergy and cost balance equations based on Specific Exergy Costing (SPECO) approach. The analysis was conducted to reveal the cost structure of the plant based on actual operating information and economic data. The exergy unitary cost of two main products of the plant, i.e., bioelectricity and biofertilizer were determined at 26.27 and 2.27 USD/GJ, respectively. The genset showed the highest overall cost rate (101.27 USD/h) followed by digester (68.41 USD/h). Furthermore, the net bioelectricity amounted to 67.81% of the overall cost rate of the products, while this value was 32.19% for both liquid and dewatered digestates. According to the results obtained, efforts should mainly focus on enhancing the efficiency of the genset in order to boost the overall performance of the system exergoeconomically. In addition, minimizing the investment-related cost of the digester could also substantially enhance the exergoeconomic performance of the plant.
This study aimed to develop an optimal continuous procedure of lipase-catalyzes transesterification of waste cooking palm oil in a packed bed reactor to investigate the possibility of large scale production further. Response surface methodology (RSM) based on central composite rotatable design (CCRD) was used to optimize the two important reaction variables packed bed height (cm) and substrate flow rate(ml/min) for the transesterification of waste cooking palm oil in a continuous packed bed reactor. The optimum condition for the transesterification of waste cooking palm oil was as follows: 10.53 cm packed bed height and 0.57 ml/min substrate flow rate. The optimum predicted fatty acid methyl ester (FAME) yield was 80.3% and the actual value was 79%. The above results shows that the RSM study based on CCRD is adaptable for FAME yield studied for the current transesterification system. The effect of mass transfer in the packed bed reactor has also been studied. Models for FAME yield have been developed for cases of reaction control and mass transfer control. The results showed very good agreement compatibility between mass transfer model and the experimental results obtained from immobilized lipase packed bed reactor operation, showing that in this case the FAME yield was mass transfer controlled.