This article presents the abilities and efficiencies of five different strains of Agrobacterium rhizogenes (strain ATCC 31798, ATCC 43057, AR12, A4 and A13) to induce hairy roots on Solanum mammosum through genetic transformation. There is significant difference in the transformation efficiency (average number of days of hairy root induction) and transformation frequency for all strains of A. rhizogenes (P < 0.05). Both A. rhizogenes strain AR12 and A13 were able to induce hairy root at 6 days of co-cultivation, which were the fastest among those tested. However, the transformation frequencies of all five strains were below 30 %, with A. rhizogenes strain A4 and A13 showing the highest, which were 21.41 ± 10.60 % and 21.43 ± 8.13 % respectively. Subsequently, the cultures for five different hairy root lines generated by five different strains of bacteria were established. However, different hairy root lines showed different growth index under the same culture condition, with the hairy root lines induced by A. rhizogenes strain ATCC 31798 exhibited largest increase in fresh biomass at 45 days of culture under 16 h light/8 h dark photoperiod in half-strength MS medium. The slowest growing hairy root line, which was previously induced by A. rhizogenes strain A13, when cultured in optimized half-strength MS medium containing 1.5 times the standard amount of ammonium nitrate and potassium nitrate and 5 % (w/v) sucrose, had exhibited improvement in growth index, that is, the fresh biomass was almost double as compared to its initial growth in unmodified half-strength MS medium.
Over the years, microalgae have been identified to be a potential source of commercially important products such as pigments, polysaccharides, polyunsaturated fatty acids and in particular, biofuels. Current demands for sustainable fuel sources and bioproducts has led to an extensive search for promising strains of microalgae for large scale cultivation. Prospective strains identified for these purposes were among others, mainly from the genera Hematococcus, Dunaliella, Botryococcus, Chlorella, Scenedesmus and Nannochloropsis. Recently, microalgae from the Selenastraceae emerged as potential candidates for biodiesel production. Strains from the Selenastraceae such as Monoraphidium sp. FXY-10, M. contortum SAG 47.80, Ankistrodesmus sp. SP2-15 and M. minutum were high biomass and lipid producers when cultivated under optimal conditions. A number of Selenastraceae strains were also reported to be suitable for cultivation in wastewater. This review highlights recent reports on potential strains from the Selenastraceae for biodiesel production and contrasts their biomass productivity, lipid productivity as well as fatty acid profile. Cultivation strategies employed to enhance their biomass and lipid productivity as well as to reduce feedstock cost are also discussed in this paper.
In this study, the effects of limited and excess nitrate on biomass, lipid production, and fatty acid profile in Messastrum gracile SE-MC4 were determined. The expression of fatty acid desaturase genes, namely stearoyl-ACP desaturase (SAD), omega-6 fatty acid desaturase (ω-6 FAD), omega-3 fatty acid desaturase isoform 1 (ω-3 FADi1), and omega-3 fatty acid desaturase isoform 2 (ω-3 FADi2) was also assessed. It was found that nitrate limitation generally increased the total oil, α-linolenic acid (C18:3n3) and total polyunsaturated fatty acid (PUFA) contents in M. gracile. The reduction of nitrate concentration from 1.76 to 0.11 mM increased the total oil content significantly from 32.5 to 41.85% (dry weight). Palmitic (C16:0) and oleic (C18:1) acids as the predominant fatty acids in this microalgae constituted between 82 and 87% of the total oil content and were relatively consistent throughout all nitrate concentrations tested. The expression of SAD, ω-6 FAD, and ω-3 FADi2 genes increased under nitrate limitation, especially at 0.11 mM nitrate. The ω-3 FADi1 demonstrated a binary up-regulation pattern of expression under both nitrate-deficient (0.11 mM) and -excess (3.55 mM) conditions. Thus, findings from this study suggested that limited or excess nitrate could be used as part of a cultivation strategy to increase oil and PUFA content following media optimisation and more efficient culture methodology. Data obtained from the expression of desaturase genes would provide valuable insights into their roles under excess and limited nitrate conditions in M. gracile, potentially paving the way for future genetic modifications.
Aerobic granular sludge (AGS) has been applied to treat a broad range of industrial and municipal wastewater. AGS can be developed in a sequencing batch reactor (SBR) with alternating anaerobic-aerobic conditions. To provide anaerobic conditions, the mixed liquor is allowed to circulate in the reactor without air supply. The circulation flow rate of mixed liquor in anaerobic condition is the most important parameter of operation in the anaerobic-AGS processes. Therefore, this study investigates the effect of circulation rate on the performance of the SBR with AGS. Two identical reactors namely R1 and R2 were operated using fermented soy sauce wastewater at circulation rate of 14.4 and 36.0 l/h, respectively. During the anaerobic conditions, the wastewater was pumped out from the upper part of the reactor and circulated back into the bottom of the reactor for 230 min. A compact and dense AGS was observed in both reactors with a similar diameter of 2.0 mm in average, although different circulation rates were adopted. The best reactor performance was achieved in R2 with chemical oxygen demand removal rate of 89%, 90% total phosphorus removal, 79% ammonia removal, 10.1 g/l of mixed liquor suspended solids and a sludge volume index of 25 ml/g.
A study has been carried out to define the effect of drastic temperature changes on the performance of lab-scale hollow-fibre MBR in treating municipal wastewater at a flux of 10 L m(-2) h(-1) (LMH). The objectives of the study were to estimate the activated sludge properties, the removal efficiencies of COD and NH(3)-N and the membrane fouling tendency under critical conditions of drastic temperature changes (23, 33, 42 & 33 °C) and MLSS concentration ranged between 6,382 and 8,680 mg/L. The study exhibited that the biomass reduction, the low sludge settleability and the supernatant turbidity were results of temperature increase. The temperature increase led to increase in SMP carbohydrate and protein, and to decrease in EPS carbohydrate and protein. The BRE of COD dropped from 80% at 23 °C to 47% at 42 °C, while the FRE was relatively constant at about 90%. Both removal efficiencies of NH(3)-N trended from about 100% at 33 °C to less than 50% at 42 °C. TMP and BWP ascended critically with temperature increase up to 336 and 304 mbar respectively by the end of the experiment. The values of suspended solids (SS) and the turbidity in the final effluent were negligible. The DO in the mixed liquor was varying with temperature change, while the pH was within the range of 6.7-8.3.
Aerobic granular sludge has a number of advantages over conventional activated sludge flocs, such as cohesive and strong matrix, fast settling characteristic, high biomass retention and ability to withstand high organic loadings, all aspects leading towards a compact reactor system. Still there are very few studies on the strength of aerobic granules. A procedure that has been used previously for anaerobic granular sludge strength analysis was adapted and used in this study. A new coefficient was introduced, called a stability coefficient (S), to quantify the strength of the aerobic granules. Indicators were also developed based on the strength analysis results, in order to categorize aerobic granules into three levels of strength, i.e. very strong (very stable), strong (stable) and not strong (not stable). The results indicated that aerobic granules grown on acetate were stronger (high density: >150 g T SSL(-1) and low S value: 5%) than granules developed on sewage as influent. A lower value of S indicates a higher stability of the granules.
Cr(VI) biosorption and bioreduction ability of locally isolated Cr-resistant bacteria was investigated using the shake-flask technique. A mixture of S. epidermidis and B. cereus showed the highest minimum inhibitory concentration (MIC) level at 750 mg/L Cr(VI) followed by S. aureus and Bacillus sp. of 250 mg/L, and A. haemolyticus of 70 mg/L. From the Langmuir adsorption isotherm, the treatment of cells with heat-acid resulted in the highest amount of Cr(VI) adsorped (78.25 mg/g dry wt. for S. epidermidis) compared to heat-acetone (67.93 mg/g dry wt. Bacillus sp.), heat only (36.05 mg/g dry wt. S. epidermidis) or untreated cells (45.40 mg/g dry wt. S. epidermidis and B. cereus). FTIR analysis showed the involvement of amine groups in Cr(VI) adsorption. In the bioreduction study, A. haemolyticus was able to completely reduce Cr(VI) up to 50 mg/L.
Aerobic granular sludge (AGS) technology has been extensively studied recently to improve sludge settling and behaviour in activated sludge systems. The main advantage is that aerobic granular sludge (AGS) can settle very fast in a reactor or clarifier because AGS is compact and has strong structure. It also has good settleability and a high capacity for biomass retention. Several experimental works have been conducted in this study to observe the settling behaviours of AGS. The study thus has two aims: (1) to compare the settling profile of AGS with other sludge flocs and (2) to observe the influence of mechanical mixing and design of the reactor to the settleability of AGS. The first experimental outcome shows that AGS settles after less than 5 min in a depth of 0.4 m compared to other sludge flocs (from sequencing batch reactor, conventional activated sludge and extended aeration) which takes more than 30 min. This study also shows that the turbulence from the mixing mechanism and shear in the reactor provides an insignificant effect on the AGS settling velocity.
Biofouling control is important for effective process of membrane bioreactor (MBR). In this study, phenomena of biofouling for immersed type extended aeration MBR with two different anti-fouling aeration intensities were studied through a laboratory set up. The objectives of this study were (a) to observe biofouling phenomena of MBR that operates under different anti-fouling bubbling intensity, and simultaneously monitors performance of the MBR in organic carbon and nutrients removal; (b) to compare effectiveness of detergent and detergent-enzyme cleaning solutions in recovering biofouled membranes that operated in the extended aeration MBR. For MBR, which operated under continuous anti-fouling aeration, deposition and accumulation of suspended biomass on membrane surface were prohibited. However, flux loss was inescapable that biofilm layer was the main problem. Membrane cleaning was successfully carried out with detergent-enzyme mixture solutions and its effectiveness was compared with result from cleaning with just detergent solution.
The objective of this study was to assess the feasibility of the Sequencing Batch Reactor (SBR) system for implementation in Malaysia. Theoretical, field, laboratory investigations, and modelling simulations have been carried out. The results of the study indicated that the SBR system was robust, relatively cost-effective, and efficient under Malaysian conditions. However, the SBR system requires highly skilled operators and continuous monitoring. This paper also attempted to identify operating conditions for the SBR system, which optimise both the removal efficiencies and the removal rates. The removal efficiencies could reach 90-96% for COD, up to 92% for TN, and 95% for SS. An approach to estimate a full operational cycle time, to estimate the de-sludging rate, and to control the biomass in the sludge has also been developed. About 4 hours react time was obtained, as 2.25 hours of nitrification with aerated slow fill and 1.75 hour of denitrification with HAc addition as an additional carbon source. Inefficient settling was one of the problems that affect the SBR effluent quality. The settling time was one hour for achieving Standard B (effluent quality) and 2 hours for Standard A.
In this study, a response surface methodology (RSM) approach using central composite design (CCD) was investigated to develop a mathematical model and to optimize the effects of pH, adsorbent amount and temperature related to the hexavalent chromium removal by biosorption on peanut shells (PSh). The highest removal percentage of 30.28% was found by the predicted model under the optimum conditions (pH of 2.11, 0.73 g of PSh and 37.2 °C) for a 100 mg/L initial Cr(VI) concentration, which was very near to the experimental value (29.92%). The PSh was characterized by SEM, EDX, FTIR, BET, XRD analyses. Moreover, a Langmuir isotherm fitted well (R2 = 0.992) with the experimental data, and the maximum adsorption capacity was discovered to be 2.48 and 3.49 mg/g respectively at 25 and 45 °C. Kinetic data were well foreseen by pseudo second order. Thermodynamic study depicted that biosorption of Cr(VI) onto PSh was spontaneous and endothermic. Regeneration of the PSh using NaOH showed a loss <5% in the Cr(VI) removal efficiency up to three recycle runs. In summary, the Cr(VI) removal onto economic, sensitive and selective biosorbent (PSh) was optimized using CCD to study biosorption behaviors.
In this study, the operational factors affecting the bioregeneration of AO7-loaded MAMS particles in batch system, namely redox condition, initial acclimated biomass concentration, shaking speed and type of acclimated biomass were investigated. The results revealed that with the use of mixed culture acclimated to AO7 under anoxic/aerobic conditions, enhancement of the bioregeneration efficiency of AO7-loaded MAMS and the total removal efficiency of COD could be achieved when the bio-decolorization and bio-mineralization stages were fully aerated with dissolved oxygen above 7 mg/L. Shorter duration of bioregeneration was achieved by using relatively higher initial biomass concentration and lower shaking speed, respectively, whereas variations of biomass concentration and shaking speed did not have a pronounced effect on the bioregeneration efficiency. The duration and efficiency of bioregeneration process were greatly affected by the chemical structures of mono-azo dyes to which the biomasses were acclimated.
Reserve lipids of microalgae are promising for biodiesel production. However, economically feasible and sustainable energy production from microalgae requires optimization of cultivation conditions for both biomass yield and lipid production of microalgae. Biomass yield and lipid production in microalgae are a contradictory problem because required conditions for both targets are different. Simultaneously, the mass cultivation of microalgae for biofuel production also depends extremely on the performance of the microalgae strains used. In this study a green unicellular microalgae Chlorella sorokiniana (DS6) isolated from the holding tanks of farm wastewater treatment plant using multi-step screening and acclimation procedures was found high-lipid producing facultative heterotrophic microalgae strain capable of growing on dairy farm effluent (DFE) for biodiesel feedstock and wastewater treatment. Morphological features and the phylogenetic analysis for the 18S rRNA identified the isolated strains. A novel three stage cultivation process of facultative strain of C. sorokiniana was examined for lipid production.
The physical characteristics, microbial activities and kinetic properties of the granular sludge biomass were investigated under the influence of different hydraulic retention times (HRT) along with the performance of the system in removal of color and COD of synthetic textile wastewater. The study was conducted in a column reactor operated according to a sequential batch reactor with a sequence of anaerobic and aerobic reaction phases. Six stages of different HRTs and different anaerobic and aerobic reaction time were evaluated. It was observed that the increase in HRT resulted in the reduction of organic loading rate (OLR). This has caused a decrease in biomass concentration (MLSS), reduction in mean size of the granules, lowered the settling ability of the granules and reduction of oxygen uptake rate (OUR), overall specific biomass growth rate (ìoverall), endogeneous decay rate (kd) and biomass yield (Yobs, Y). When the OLR was increased by adding carbon sources (glucose, sodium acetate and ethanol), there was a slight increase in the MLSS, the granules mean size, ìoverall, and biomass yield. Under high HRT, increasing the anaerobic to aerobic reaction time ratio caused an increase in the concentration of MLSS, mean size of granules and lowered the SVI value and biomass yield. The ìoverall and biomass yield increased with the reduction in anaerobic/aerobic time ratio. The HRT of 24 h with anaerobic and aerobic reaction time of 17.8 and 5.8 h respectively appear to be the best cycle operation of SBR. Under these conditions, not only the physical properties of the biogranules have improved, the highest removal of color (i.e. 94.1±0.6%) and organics (i.e. 86.5±0.5%) of the synthetic textile dyeing wastewater have been achieved.
Microbial granular sludge that is capable to treat textile wastewater in a single reactor under intermittent anaerobic and aerobic conditions was developed in this study. The granules were cultivated using mixed sewage and textile mill sludge in combination with anaerobic granules collected from an anaerobic sludge blanket reactor as seed. The granules were developed in a single sequential batch reactor (SBR) system under alternating anaerobic and aerobic condition fed with synthetic textile wastewater. The characteristics of the microbial granular sludge were monitored throughout the study period. During this period, the average size of the granules increased from 0.02 +/- 0.01 mm to 2.3 +/- 1.0 mm and the average settling velocity increased from 9.9 +/- 0.7 m h(-1) to 80 +/- 8 m h(-1). This resulted in an increased biomass concentration (from 2.9 +/- 0.8 g L(-1) to 7.3 +/- 0.9 g L(-1)) and mean cell residence time (from 1.4 days to 8.3 days). The strength of the granules, expressed as the integrity coefficient also improved. The sequential batch reactor system demonstrated good removal of COD and ammonia of 94% and 95%, respectively, at the end of the study. However, only 62% of color removal was observed. The findings of this study show that granular sludge could be developed in a single reactor with an intermittent anaerobic-aerobic reaction phase and is capable in treating the textile wastewater.
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 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.
Dairy cattle treated wastewaters are potential resources for production of microalgae biofuels. A study was conducted to evaluate the capability of Arthrospira platensis cultivated in dairy farm wastewater for biodiesel production. The biomass of Arthrospira platensis was found to be 4.98 g L-1 and produced 30.23 wt% lipids to dry biomass cultivated in wastewater which was found nitrogen stressed in photo bioreactor. The extracted lipid displayed a suitable fatty acid profile for biodiesel, although the content of linolenic acid was found a little higher than the standard EN14214. It was found that nitrogen stressed medium increase the total lipid content but temperature and intensities of light were the most important factors to control the quantity of linolenic acid and hence the quality of biodiesel, while the optimum CO2 helped to achieve maximum biomass and triacylglycerols. The Arthrospira platensis offer a good option for the treatment of wastewater before final discharge.
Hermetia illucens larvae by nature are a decomposer which fed on organic wastes. This study explores the potential of producing biodiesel using lipids from H. illucens larvae. Three types of organic wastes (sewage sludge, fruit waste and palm decanter cake from oil palm mill) were selected based on considerable generation and disposal concern in the area of study as well as lack of investigations as feed for Hermetia illucens larvae in current literatures. Growth rate of the larvae was determined with studying the changes in the biomass per day. H. illucens larvae fed with fruit waste and palm decanter cake have shown growth rates of 0.52±0.02 and 0.23±0.09 g d(-1), respectively. No positive sign of growth were observed in the larvae fed with treated sewage sludge (-0.04±0.01 g d(-1)). Biodiesel as fatty acid methyl ester (FAME) was synthesized by transesterification of the larvae lipid using sulphuric acid as catalyst in methanol. FAME produced was ascertained using ATR-FTIR spectroscopy and GC-MS. The main compositions of fatty acid were found to be C12:0, C16:0 and C18:1n9c. Fatty acid composition of C12:0 fed with fruit waste, sewage sludge and palm decanter was found to be most abundant in the larvae lipid. The amount of C12:0 obtained was 76.13%, 58.31% and 48.06%, respectively. In addition, fatty acid of C16:0 was attained at 16.48% and 25.48% fed with sewage sludge and palm decanter, respectively. Based on the findings, FAME derived from larvae lipids is feasible to be used for biodiesel production.
Microwave assisted acid hydrolysis (H2SO4 and HCl with >0.5 mol/L) to produce bioethanol from sago pith waste (SPW) was studied. The energy consumption for microwave hydrolysis at different energy inputs and acid concentration were calculated. The overall energy consumption for bioethanol fuel production from SPW was assessed. A maximum of 88% glucose yield and 80% ethanol yield (3.1 g ethanol per 10 g SPW) were obtained using 1.0 mol/L H2SO4. Microwave hydrolysis using 1.0 mol/L H2SO4 consumed the minimum energy of 8.1 kJ to produce 1 g glucose from SPW when energy input was fixed at 54 kJ (900 W for 1 min). In general, 1 g glucose can produce 16 kJ. The overall energy consumption for fuel grade bioethanol production from SPW was 31.77 kJ per g ethanol, which was slightly higher than the lower heating values of ethanol (26.74 kJ/g ethanol).