The objective of this study is to investigate the effect of salts addition to fermentation medium on hydrogen production, under anaerobic batch culture system. In this study, batch experiments were conducted to investigate the inhibitory effect of both NaCl and sodium acetate on hydrogen production. The optimum pH and temperature for hydrogen production were at initial pH of 7.0 and 30 degrees C. Enhanced production of hydrogen, using glucose as substrate was achieved. In the absence of Sodium Chloride and Sodium Acetate enhanced hydrogen yield (Y(P/S)) from 350 mL g(-1) glucose utilized to 391 mL g(-1) glucose utilized with maximum hydrogen productivity of 77.5 ml/L/h. Results also show that sodium chloride and sodium acetate in the medium adversely affect growth. Hydrogen yield per biomass (Y(P/X)) of 254 ml/L/g, biomass per substrate utilized (Y(X/S)) of 0.268 and (Y(H2/S) of 0.0349. The results suggested that Sodium at any concentration resulted to inhibit the bacterial productivity of hydrogen.
Composting of Palm Oil Mill Sludge (POMS) with sawdust was conducted in natural aerated reactor. Composting using natural aerated reactor is cheap and simple. The goal of this study is to observe the potential of composting process and utilizing compost as media for growing Cymbopogun citratus, one of Malaysia herbal plant. The highest maximum temperature achieved is about 40 degrees C and to increase temperature bed, more biodegradable substrate needs to be added. The pH value decrease along the process with final pH compost is acidic (pH 5.7). The highest maximum organic losses are about 50% with final C/N ratio of the compost is about 19. Final compost also showed some fertilizing value but need to be adjusted to obtain an ideal substrate. Addition of about 70% sandy soil causes highest yield and excellent root development for C. citratus in potted media. Beside that, compost from POMS-sawdust also found to have fertilizer value and easy to handle. Composting of POMS with sawdust shows potential as an alternative treatment to dispose and recycle waste components.
Biological kinetic (bio-kinetic) study of the anaerobic stabilization pond treatment of palm oil mill effluent (POME) was carried out in a laboratory anaerobic bench scale reactor (ABSR). The reactor was operated at different feed flow-rates of 0.63, 0.76, 0.95, 1.27, 1.9 and 3.8l of raw POME for a day. Chemical oxygen demand (COD) as influent substrates was selected for bio-kinetic study. The investigation showed that the growth yield (Y(G)), specific biomass decay (b), maximum specific biomass growth rate (mu(max)), saturation constant (K(s)) and critical retention time (Theta(c)) were in the range of 0.990 g VSS/g COD(removed) day, 0.024 day(-1), 0.524 day(-1), 203.433 g COD l(-1) and 1.908 day, respectively.
The production of highly polluting palm oil mill effluent (POME) has resulted in serious environmental hazards. While anaerobic digestion is widely accepted as an effective method for the treatment of POME, anaerobic treatment of POME alone has difficulty meeting discharge limits due to the high organic strength of POME. Hence, subsequent post-treatment following aerobic treatment is vital to meet the discharge limits. The objective of the present study is to investigate the aerobic treatment of anaerobically digested POME by using a sequencing batch reactor (SBR). The SBR performance was assessed by measuring Chemical Oxygen Demand (COD), Biochemical Oxygen Demand (BOD) and Total Suspended Solids (TSS) removal as well as Sludge Volume Index (SVI). The operating pH and dissolved oxygen concentrations were found to be 8.25-9.14 and 1.5-6.4 mg/L, respectively, throughout the experiment. The experimental results demonstrate that MLVSS, OLR and sludge loading rate (SLR) play a significant role in the organic removal efficiency of SBR systems and therefore, further investigation on these parameters was conducted to attain optimum SBR performance. Maximum COD (95-96%), BOD (97-98%) and TSS (98-99%) removal efficiencies were achieved at optimum OLR, SLR and MLVSS concentration ranges of 1.8-4.2 kg COD/m(3)day, 2.5-4.6 kg TSS/m(3)day and 22,000-25,000 mg/L, respectively. The effluent quality remained stable and complied with the discharge limit. At the same time, the sludge showed good settling properties with average SVI of 65. It is envisaged that the SBR process could complement the anaerobic treatment to produce final treated effluent which meets the discharge limit.
In this study treatment of palm oil mill effluent (POME) was investigated using aerobic oxidation based on an activated sludge process. The effects of sludge volume index, scum index and mixed liquor suspended solids during the acclimatizing phase and biomass build-up phase were investigated in order to ascertain the reactor stability. The efficiency of the activated sludge process was evaluated by treating anaerobically digested and diluted raw POME obtained from Golden Hope Plantations, Malaysia. The treatment of POME was carried out at a fixed biomass concentration of 3900+/-200mg/L, whereas the corresponding sludge volume index was found to be around 105+/-5mL/g. The initial studies on the efficiency of the activated sludge reactor were carried out using diluted raw POME for varying the hydraulic retention time, viz: 18, 24, 30 and 36h and influent COD concentration, viz: 1000, 2000, 3000, 4000 and 5000mg/L, respectively. The results showed that at the end of 36h of hydraulic retention time for the above said influent COD, the COD removal efficiencies were found to be 83%, 72%, 64%, 54% and 42% whereas at 24h hydraulic retention time they were 57%, 45%, 38%, 30% and 27%, respectively. The effectiveness of aerobic oxidation was also compared between anaerobically digested and diluted raw POME having corresponding CODs of 3908 and 3925mg/L, for varying hydraulic retention time, viz: 18, 24, 30, 36, 42, 48, 54 and 60h. The dissolved oxygen concentration and pH in the activated sludge reactor were found to be 1.8-2.2mg/L and 7-8.5, respectively. The scum index was found to rise from 0.5% to 1.9% during the acclimatizing phase and biomass build-up phase.
Box-Wilson design (BWD) model was applied to determine the optimum values of influencing parameters in anaerobic fermentation to produce hydrogen using Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). The main focus of the study was to find the optimal relationship between the hydrogen yield and three variables including initial substrate concentration, initial medium pH and reaction temperature. Microbial growth kinetic parameters for hydrogen production under anaerobic conditions were determined using the Monod model with incorporation of a substrate inhibition term. The values of micro(max) (maximum specific growth rate) and K, (saturation constant) were 0.398 h(-1) and 5.509 g L(-1), respectively, using glucose as the substrate. The experimental substrate and biomass-concentration profiles were in good agreement with those obtained by the kinetic-model predictions. By varying the conditions of the initial substrate concentration (1-40 g L(-1)), reaction temperature (25-40 degrees C) and initial medium pH (4-8), the model predicted a maximum hydrogen yield of 3.24 mol H2 (mol glucose)(-1). The experimental data collected utilising this design was successfully fitted to a second-order polynomial model. An optimum operating condition of 10 g L(-1) initial substrate concentration, 37 degrees C reaction temperature and 6.0 +/- 0.2 initial medium pH gave 80% of the predicted maximum yield of hydrogen where as the experimental yield obtained in this study was 77.75% exhibiting a close accuracy between estimated and experimental values. This is the first report to predict bio-hydrogen yield by applying Box-Wilson Design in anaerobic fermentation while optimizing the effects of environmental factors prevailing there by investigating the effects of environmental factors.
Foaming problem which occurred occasionally during food waste (FW) anaerobic digestion (AD) was investigated with the Malaysian FW by stepwise increase in organic loading (OL) from 0.5 to 7.5 g VS/L. The FW feedstock with carbon to nitrogen (C/N) ratio of 17 was upgraded to C/N ratio of 26 and 30 by mixing with other wastes. The digestion which was carried out at 37 °C in 1-L batch reactors showed that foam formation initiated at OL of 1.5 g VS/L and was further enhanced as OL of feedstock was increased. The digestion foaming reached its maximum at OL of 5.5 g VS/L and did not increase further even when OL was increased to 7.5 g VS/Ld. Increase in the C/N ratio of feedstock significantly enhanced the microbial degradation activity, leading to better removal of foam causing intermediates and reduced foaming in the reactor by up to 60%.
Influence of the labile organic fraction (LOF) on anaerobic digestion of food waste was investigated in different S/I ratio of 0.33, 0.5, 1.0, 2.0 and 4.0g-VSsubstrate/g-VSinoculum. Two types of substrate, standard food waste (Substrate 1) and standard food waste with the supernatant (containing LOF) removed (Substrate 2) were used. Highest methane yield of 435ml-CH4g-VS(-1) in Substrate 1 was observed in the lowest S/I ratio, while the methane yield of the other S/I ratios were 38-73% lower than the highest yield due to acidification. The methane yields in Substrate 2 were relatively stable in all S/I conditions, although the maximum methane yield was low compared with Substrate 1. These results showed that LOF in food waste causes acidification, but also contributes to high methane yields, suggesting that low S/I ratio (<0.33) is required to obtain a reliable methane yield from food waste compared to other organic substrates.
This study was aimed to investigate the production of methane gas from three different types of food waste (vegetables waste, fruit waste and grain waste) using batch type anaerobic digestion method. The digestion process was conducted by using temperature range of 27 to 36 degrees C and pH 6.5 to 7.5 to yield an optimum condition for the digestion process. The digestion was continued for a period of two weeks with the aid of cow dung as the inoculums. It was found that the grain waste yielded the highest methane 2546 mL due to the high content of carbohydrate. At the mean time, the fruit waste produced the second highest methane gas with 2000 mL as well as the vegetable waste generated the lowest methane gas with volume of 1468 mL. The vegetable waste produced the lowest methane gas because the vegetables waste contains high fibres and cellulose walls but low in glucose amount. For the fertilization test, fruit waste demonstrated the best observation for the growth of plant due to high content of potassium and followed by vegetable waste. The least effective fertilizer was grain waste due to less content of nutrients essential for plants growth.
The effect of Leucaena leucocephala hybrid-Bahru (LLB), which contains a high concentration of condensed tannins, on cellulolytic rumen fungal population in goats was investigated using real-time PCR. The fungal population in goats fed LLB was inhibited during the first 10 days of feeding, but after 15 days of feeding, there was a tremendous increase of fungal population (157.0 μg/ml), which was about fourfold more than that in control goats (39.7 μg/ml). However, after this period, the fungal population decreased continuously, and at 30 days of feeding, the fungal population (50.6 μg/ml) was not significantly different from that in control goats (55.4 μg/ml).
Recently, soybean tempeh has received great attention due to many advantages such as higher nutritional value, lower production cost, and shorter fermentation time. In this study, the in vivo hepatoprotective and antioxidant effects of nutrient enriched soybean tempeh (NESTE) were determined. NESTE fermentation process which involved anaerobic incubation was previously proclaimed to increase the content of amino acids and antioxidant properties remarkably. The evaluation of histological sections, serum biochemical markers (aspartate aminotransferase (AST), alanine aminotransferase (ALT), and cholesterol and triglycerides (TG)), liver immune response level (nitric oxide (NO)) and liver antioxidant level (superoxide dismutase (SOD), ferric reducing antioxidant power (FRAP), and malondialdehyde (MDA)) was conducted in order to compare the effects of nonfermented soybean extract (SBE) and fermented soybean extract (NESTE) on alcohol-induced liver damage in mice. Results demonstrated that 1000 mg/kg of NESTE can significantly reduce the levels of AST, ALT, cholesterol, TG, MDA, and NO. On the other hand, it also raised the level of SOD and FRAP. Furthermore, the histological examination on 1000 mg/kg NESTE treatment group showed that this extract was capable of recovering the damaged hepatocytes to their normal structures. Thus, it can be concluded that NESTE produced through fermentation process was able to enhance hepatoprotective and antioxidant effects in vivo.
The metabolic role of 6-phosphogluconate dehydrogenase (gnd) under anaerobic conditions with respect to succinate production in Escherichia coli remained largely unspecified. Herein we report what are to our knowledge the first metabolic gene knockout of gnd to have increased succinic acid production using both glucose and glycerol substrates in E. coli. Guided by a genome scale metabolic model, we engineered the E. coli host metabolism to enhance anaerobic production of succinic acid by deleting the gnd gene, considering its location in the boundary of oxidative and non-oxidative pentose phosphate pathway. This strategy induced either the activation of malic enzyme, causing up-regulation of phosphoenolpyruvate carboxylase (ppc) and down regulation of phosphoenolpyruvate carboxykinase (ppck) and/or prevents the decarboxylation of 6 phosphogluconate to increase the pool of glyceraldehyde-3-phosphate (GAP) that is required for the formation of phosphoenolpyruvate (PEP). This approach produced a mutant strain BMS2 with succinic acid production titers of 0.35gl(-1) and 1.40gl(-1) from glucose and glycerol substrates respectively. This work further clearly elucidates and informs other studies that the gnd gene, is a novel deletion target for increasing succinate production in E. coli under anaerobic condition using glucose and glycerol carbon sources. The knowledge gained in this study would help in E. coli and other microbial strains development for increasing succinate production and/or other industrial chemicals.
This study assessed the efficacy of granular cylindrical periodic discontinuous batch reactors (GC-PDBRs) for produced water (PW) treatment by employing eggshell and waste activated sludge (WAS) derived Nickel (Ni) augmented biochar. The synthesized biochar was magnetized to further enhance its contribution towards achieving carbon neutrality due to carbon negative nature, Carbon dioxide (CO2) sorption, and negative priming effects. The GC-PDBR1 and GC-PDBR2 process variables were optimized by the application of central composite design (CCD). This is to maximize the decarbonization rate. Results showed that the systems could reduce total phosphorus (TP) and chemical oxygen demand (COD) by 76-80% and 92-99%, respectively. Optimal organic matter and nutrient removals were achieved at 80% volumetric exchange ratio (VER), 5 min settling time and 3000 mg/L mixed liquor suspended solids (MLSS) concentration with desirability values of 0.811 and 0.954 for GC-PDBR1 and GC-PDBR2, respectively. Employing four distinct models, the biokinetic coefficients of the GC-PDBRs treating PW were calculated. The findings indicated that First order (0.0758-0.5365) and Monod models (0.8652-0.9925) have relatively low R2 values. However, the Grau Second-order model and Modified Stover-Kincannon model have high R2 values. This shows that, the Grau Second Order and Modified Stover-Kincannon models under various VER, settling time, and MLSS circumstances, are more suited to explain the removal of pollutants in the GC-PDBRs. Microbiological evaluation demonstrated that a high VER caused notable rises in the quantity of several microorganisms. Under high biological selective pressure, GC-PDBR2 demonstrated a greater percentage of nitrogen removal via autotrophic denitrification and a greater number of nitrifying bacteria. The overgrowth of bacteria such as Actinobacteriota spp. Bacteroidota spp, Gammaproteobacteria, Desulfuromonas Mesotoga in the phylum, class, and genus, has positively impacted on granule formation and stability. Taken together, our study through the introduction of intermittent aeration GC-PDBR systems with added magnetized waste derived biochar, is an innovative approach for simultaneous aerobic sludge granulation and PW treatment, thereby providing valuable contributions in the journey toward achieving decarbonization, carbon neutrality and sustainable development goals (SDGs).
Despite efforts to address the composition of the microbial community during the anaerobic treatment of palm oil mill effluent (POME), its composition in relation to biodegradation in the full-scale treatment system has not yet been extensively examined. Therefore, a thorough analysis of bacterial and archaeal communities was performed in the present study using MiSeq sequencing at the different stages of the POME treatment, which comprised anaerobic as well as facultative anaerobic and aerobic processes, including the mixed raw effluent (MRE), mixing pond, holding tank, and final discharge phases. Based on the results obtained, the following biodegradation processes were suggested to occur at the different treatment stages: (1) Lactobacillaceae (35.9%) dominated the first stage, which contributed to high lactic acid production; (2) the higher population of Clostridiaceae in the mixing pond (47.7%) and Prevotellaceae in the holding tank (49.7%) promoted acetic acid production; (3) the aceticlastic methanogen Methanosaetaceae (0.6-0.8%) played a role in acetic acid degradation in the open digester and closed reactor for methane generation; (4) Syntrophomonas (21.5-29.2%) appeared to be involved in the degradation of fatty acids and acetic acid by syntrophic cooperation with the hydrogenotrophic methanogen, Methanobacteriaceae (0.6-1.3%); and (5) the phenols and alcohols detected in the early phases, but not in the final discharge phase, indicated the successful degradation of lignocellulosic materials. The present results contribute to a better understanding of the biodegradation mechanisms involved in the different stages of the full-scale treatment of POME.
This study determined the effect of light intensity and photoperiod on the dry cell weight and total amount of carotenoids in four isolates of purple non-sulfur bacteria obtained from shaded and exposed microhabitats of a mangrove ecosystem in Kota Kinabalu, Sabah, Malaysia. The initial isolation of the bacteria was carried out using synthetic 112 medium under anaerobic conditions (2.5 klx) at 30 ± 2°C. On the basis of colony appearance, cell morphology, gram staining, motility test, and 16S rRNA gene sequencing analyses, all four bacteria were identified as Afifella marina. One of the bacterial isolates, designated as Af. marina strain ME, which was extracted from an exposed mud habitat within the mangrove ecosystem, showed the highest yield in dry cell weight (4.32± 0.03 g/l) as well as total carotenoids (0.783 ± 0.002 mg/g dry cell weight). These values were significantly higher than those for dry cell weight (3.77 ± 0.02g/l ) and total carotenoid content (0.706 ± 0.008 mg/g) produced by the isolates from shaded habitats. Further analysis of the effect of 10 levels of light intensity on the growth characteristics of Af. marina strain ME showed that the optimum production of dry cell weight and total carotenoids was achieved at different light intensities and incubation periods. The bacterium produced the highest dry cell weight of 4.98 g/l at 3 klx in 72 h incubation, but the carotenoid production of 0.783 mg/g was achieved at 2.5 klx in 48 h incubation. Subsequent analysis of the effect of photoperiod on the production of dry cell weight and total carotenoids at optimum light intensities (3 and 2.5 klx, respectively) revealed that 18 and 24 h were the optimum photoperiods for the production of dry cell weight and total carotenoids, respectively. The unique growth characteristics of the Af. marina strain ME can be exploited for biotechnology applications.
The biodegradation characteristics of palm oil mill effluent (POME) and the related microbial community were studied in both actual sequential anaerobic ponds in Malaysia and enrichment cultures. The significant degradation of the POME was observed in the second pond, in which the temperature was 35-37 °C. In this pond, biodegradation of major long chain fatty acids (LCFA), such as palmitic acid (C16:0) and oleic acid (C18:1), was also confirmed. The enrichment culture experiment was conducted with different feeding substrates, i.e. POME, C16:0 and C18:1, at 35 °C. Good recovery of methane indicated biodegradation of feeds in the POME and C16:0 enrichments. The methane production rate of the C18:1 enrichment was slower than other substrates and inhibition of methanogenesis was frequently observed. Denaturing gradient gel electrophoresis (DGGE) analyses indicated the existence of LCFA-degrading bacteria, such as the genus Syntrophus and Syntorophomonas, in all enrichment cultures operated at 35 °C. Anaerobic degradation of the POME under mesophilic conditions was stably processed as compared with thermophilic conditions.
The degradation of (RS)-MCPP was investigated in an anaerobic membrane bioreactor (AnMBR) using nitrate as an available electron acceptor under different COD/NO(3)(-)-N ratios. Results showed high soluble COD removal efficiency (80-93%) when the reactor was operated at high COD/NO(3)(-)-N ratios. However, the COD removal started to decline (average 15%) at high nitrate concentrations coinciding with a drop in nitrate removal efficiency to 37%, suggesting that the denitrification activity dropped and affected the AnMBR performance when nitrate was the predominant electron acceptor. Additionally, the removal efficiency of (RS)-MCPP increased from 2% to 47% with reducing COD/NO(3)(-)-N ratios, whilst the (RS)-MCPP specific utilisation rate (SUR) was inversely proportional to the COD/NO(3)(-)-N ratio, suggesting that a lower COD/NO(3)(-)-N ratios had a positive influence on the (RS)-MCPP SUR. Although nitrate had a major impact on methane production rates, the methane composition was stable (approximately 80%) for COD/NO(3)(-)-N ratios of 23 or more.
A novel wastewater treatment system consisting of an up-flow anaerobic sludge blanket (UASB) reactor and a down-flow hanging sponge (DHS) reactor with sulfur-redox reaction was developed for treatment of municipal sewage under low-temperature conditions. In the UASB reactor, a novel phenomenon of anaerobic sulfur oxidation occurred in the absence of oxygen, nitrite and nitrate as electron acceptors. The microorganisms involved in anaerobic sulfur oxidation have not been elucidated. Therefore, in this study, we studied the microbial communities existing in the UASB reactor that probably enhanced anaerobic sulfur oxidation. Sludge samples collected from the UASB reactor before and after sulfur oxidation were used for cloning and terminal restriction fragment length polymorphism (T-RFLP) analysis of the 16S rRNA genes of the bacterial and archaeal domains. The microbial community structures of bacteria and archaea indicated that the genus Smithella and uncultured bacteria within the phylum Caldiserica were the dominant bacteria groups. Methanosaeta spp. was the dominant group of the domain archaea. The T-RFLP analysis, which was consistent with the cloning results, also yielded characteristic fingerprints for bacterial communities, whereas the archaeal community structure yielded stable microbial community. From these results, it can be presumed that these major bacteria groups, genus Smithella and uncultured bacteria within the phylum Caldiserica, probably play an important role in sulfur oxidation in UASB reactors.
To date, little information is known about the operation of the enhanced biological phosphorus removal (EBPR) process in tropical climates. Along with the global concerns on nutrient pollution and the increasing array of local regulatory requirements, the applicability and compliance accountability of the EBPR process for sewage treatment in tropical climates is being evaluated. A sequencing batch reactor (SBR) inoculated with seed sludge from a conventional activated sludge (CAS) process was successfully acclimatized to EBPR conditions at 28 °C after 13 days' operation. Enrichment of Candidatus Accumulibacter phosphatis in the SBR was confirmed through fluorescence in situ hybridization (FISH). The effects of operational pH and influent C:P ratio on EBPR were then investigated. At pH 7 or pH 8, phosphorus removal rates of the EBPR processes were relatively higher when operated at C:P ratio of 3 than C:P ratio of 10, with 0.019-0.020 and 0.011-0.012 g-P/g-MLVSS•day respectively. One-year operation of the 28 °C EBPR process at C:P ratio of 3 and pH 8 demonstrated stable phosphorus removal rate of 0.020 ± 0.003 g-P/g-MLVSS•day, corresponding to effluent with phosphorus concentration <0.5 mg/L. This study provides the first evidence on good EBPR activity at relatively high temperature, indicating its applicability in a tropical climate.
Enterococcus sp. strain C1 is a facultative anaerobe which was coisolated with Citrobacter sp. strain A1 from a sewage oxidation pond. Strain C1 could degrade azo dyes very efficiently via azo reduction and desulfonation in a microaerophilic environment. Here the draft genome sequence of Enterococcus sp. C1 is reported.