The composting of lignocellulosic oil palm empty fruit bunch (OPEFB) with continuous addition of palm oil mill (POME) anaerobic sludge which contained nutrients and indigenous microbes was studied. In comparison to the conventional OPEFB composting which took 60-90 days, the rapid composting in this study can be completed in 40 days with final C/N ratio of 12.4 and nitrogen (2.5%), phosphorus (1.4%), and potassium (2.8%), respectively. Twenty-seven cellulolytic bacterial strains of which 23 strains were closely related to Bacillus subtilis, Bacillus firmus, Thermobifida fusca, Thermomonospora spp., Cellulomonas sp., Ureibacillus thermosphaericus, Paenibacillus barengoltzii, Paenibacillus campinasensis, Geobacillus thermodenitrificans, Pseudoxanthomonas byssovorax which were known as lignocellulose degrading bacteria and commonly involved in lignocellulose degradation. Four isolated strains related to Exiguobacterium acetylicum and Rhizobium sp., with cellulolytic and hemicellulolytic activities. The rapid composting period achieved in this study can thus be attributed to the naturally occurring cellulolytic and hemicellulolytic strains identified.
Periphyton dominated by the cellulose-rich filamentous green alga Cladophora forms conspicuous growths along rocky marine and freshwater shorelines worldwide, providing habitat for diverse epibionts. Bacterial epibionts have been inferred to display diverse functions of biogeochemical significance: N-fixation and other redox reactions, phosphorus accumulation, and organic degradation. Here, we report taxonomic diversity of eukaryotic and prokaryotic epibionts and diversity of genes associated with materials cycling in a Cladophora metagenome sampled from Lake Mendota, Dane Co., WI, USA, during the growing season of 2012. A total of 1,060 distinct 16S, 173 18S, and 351 28S rRNA operational taxonomic units, from which >220 genera or species of bacteria (~60), protists (~80), fungi (6), and microscopic metazoa (~80), were distinguished with the use of reference databases. We inferred the presence of several algal taxa generally associated with marine systems and detected Jaoa, a freshwater periphytic ulvophyte previously thought endemic to China. We identified six distinct nifH gene sequences marking nitrogen fixation, >25 bacterial and eukaryotic cellulases relevant to sedimentary C-cycling and technological applications, and genes encoding enzymes in aerobic and anaerobic pathways for vitamin B12 biosynthesis. These results emphasize the importance of Cladophora in providing habitat for microscopic metazoa, fungi, protists, and bacteria that are often inconspicuous, yet play important roles in ecosystem biogeochemistry.
This paper critically reviews the impacts of supplementing trace elements on the anaerobic digestion performance. The in-depth knowledge of trace elements as micronutrients and metalloenzyme components justifies trace element supplementation into the anaerobic digestion system. Most of the earlier studies reported that trace elements addition at (sub)optimum dosages had positive impacts mainly longer term on digester stability with greater organic matter degradation, low volatile fatty acids (VFA) concentration and higher biogas production. However, these positive impacts and element requirements are not fully understood, they are explained on a case to case basis because of the great variance of the anaerobic digestion operation. Iron (Fe), nickel (Ni) and cobalt (Co) are the most studied and desirable elements. The right combination of multi-elements supplementation can have greater positive impact. This measure is highly recommended, especially for the mono-digestion of micronutrient-deficient substrates. The future research should consider the aspect of trace element bioavailability.
Large volumes of untreated palm oil mill effluent (POME) pose threat to aquatic environment due to the presence of very high organic content. The present investigation involved two pilot-scale anaerobic expanded granular sludge bed (EGSB) reactors, continuously operated for 1 year to treat POME. Setting HRT at 9.8 d, the anaerobic EGSB reactors reduced COD from 71179 mg/L to 12341 mg/L and recycled half of sludge by a dissolved air flotation (DAF). The average effluent COD was 3587 mg/L with the consistent COD removal efficiency of 94.89%. Adding cationic polymer (PAM) dose of 30 mg/L to DAF unit and recycling its half of sludge caused granulation of anaerobic sludge. Bacilli and small coccid bacteria were the dominant microbial species of the reactor. The reactor produced 27.65 m(3) of biogas per m(3) of POME which was utilized for electricity generation.
Treatment of ammonia- and nitrate-rich wastewater, such as that generated in the aquaculture industry, is important to prevent environmental pollution. The anaerobic ammonium oxidation (anammox) process has been reported as a great alternative in reducing ammoniacal nitrogen concentration in aquaculture wastewater treatment compared to conventional treatment systems. This paper will highlight the impact of the anammox process on aquaculture wastewater, particularly in the regulation of ammonia and nitrogen compounds. The state of the art for anammox treatment systems is discussed in comparison to other available treatment methods. While the anammox process is viable for the treatment of aquaculture wastewater, the efficiency of nitrogen removal could be further improved through the proper use of anammox bacteria, operating conditions, and microbial diversity. In conclusion, a new model of the anammox process is proposed in this review.
The world currently obtains its energy from the fossil fuels such as oil, natural gas and coal. However, the international crisis in the Middle East, rapid depletion of fossil fuel reserves as well as climate change have driven the world towards renewable energy sources which are abundant, untapped and environmentally friendly. Malaysia has abundant biomass resources generated from the agricultural industry particularly the large commodity, palm oil. This paper will focus on palm oil mill effluent (POME) as the source of renewable energy from the generation of methane and establish the current methane emission from the anaerobic treatment facility. The emission was measured from two anaerobic ponds in Felda Serting Palm Oil Mill for 52 weeks. The results showed that the methane content was between 35.0% and 70.0% and biogas flow rate ranged between 0.5 and 2.4 L/min/m(2). Total methane emission per anaerobic pond was 1043.1 kg/day. The total methane emission calculated from the two equations derived from relationships between methane emission and total carbon removal and POME discharged were comparable with field measurement. This study also revealed that anaerobic pond system is more efficient than open digesting tank system for POME treatment. Two main factors affecting the methane emission were mill activities and oil palm seasonal cropping.
Human gastrointestinal microbiota (HGIM) incorporate a large number of microbes from different species. Anaerobic bacteria are the dominant organisms in this microbial consortium and play a crucial role in human health. In addition to their functional role as the main source of many essential metabolites for human health, they are considered as biotherapeutic agents in the regulation of different human metabolites. They are also important in the prevention and in the treatment of different physical and mental diseases. Bifidobacteria are the dominant anaerobic bacteria in HGIM and are widely used in the development of probiotic products for infants, children and adults. To develop bifidobacteria-based bioproducts, therefore, it is necessary to develop a large-scale biomass production platform based on a good understanding of the ideal medium and bioprocessing parameters for their growth and viability. In addition, high cell viability should be maintained during downstream processing and storage of probiotic cell powder or the final formulated product. In this work we review the latest information about the biology, therapeutic activities, cultivation and industrial production of bifidobacteria.
Biological treatments, such as activated sludge process, are common methods to treat municipal and industrial wastewaters. However, they produce huge amounts of waste activated sludge (WAS). The excess sludge treatment and disposal are a challenge for wastewater treatment plants due to economic, environmental, and regulatory factors. In this study, photo-Fenton pretreatment (oxidation using hydrogen peroxide and iron catalyst aided with UV light) was optimized using response surface methodology (RSM) and central composite design (CCD) to determine the effects of three operating parameters (H2O2 dosage, H2O2/Fe2+ molar ratio, and irradiation time) on disintegration and dewaterability of WAS. MLVSS removal, capillary suction time (CST) reduction, sCOD, and EPS were obtained as 70%, 25%, 12,000 mg/L, and 500 mg/L, respectively, at the optimal conditions, i.e., 725 g H2O2/kg TS, H2O2/Fe2+ molar ratio 80, and irradiation time 40 min. Two batch-fed completely mixed mesophilic anaerobic digesters were then operated at 15-day solid retention time (SRT) and 37 ± 0.5 °C to compare the digestibility of untreated and photo-Fenton pretreated sludge in terms of volatile solids (VS) reduction, COD removal, and biogas production at steady-state operations. Photo-Fenton pretreatment followed by anaerobic digestion of WAS was very effective and yielded 75.7% total VS reduction, 81.5% COD removal, and 0.29-0.31 m3/kg VSfed·d biogas production rate, compared to 40.7% total VS solid reduction, 54.7% COD removal, and 0.12-0.17 m3/kg VSfed·d biogas production rate for control. Thus, photo-Fenton can be a useful pretreatment step in sludge management.
Hydrazine is an intermediate product of the anaerobic ammonium oxidation (Anammox) process where both ammonium and nitrite in wastewater are converted to nitrogen gas by bacteria. In this study the effect of external hydrazine addition (5, 10, 15, and 20 mg/L) on the start-up period of the Anammox process was studied using sequencing batch reactors (SBRs). The SBR with an addition of 10 mg/L hydrazine took only 7 weeks to stabilize and achieve the maximum removal of ammonium and nitrite, whereas the SBR without the addition of hydrazine took 12 weeks. The amount of Heme C extracted from the biomass indicated that externally added hydrazine accelerated the growth of Anammox bacteria and reduced the release of nitrous oxide gas from the reactors.
Sago hampas is a starch-based biomass from sago processing industries consisted of 58% remaining starch. This study has demonstrated the bioconversion of sago hampas to volatile fatty acids (VFAs) by Clostridium beijerinckii SR1 via anaerobic digestion. Higher total VFAs were obtained from sago hampas (5.04 g/L and 0.287 g/g) as compared to commercial starch (5.94 g/L and 0.318 g/g). The physical factors have been investigated for the enhancement of VFAs production using one-factor-at-a-time (OFAT). The optimum condition; 3% substrate concentration, 3 g/L of yeast extract concentration and 2 g/L of ammonium nitrate enhanced the production of VFAs by 52.6%, resulted the total VFAs produced is 7.69 g/L with the VFAs yield of 0.451 g/g. VFAs hydrolysate produced successfully generated 273.4 mV of open voltage circuit and 61.5 mW/m2 of power density in microbial fuel cells. It was suggested that sago hampas provide as an alternative carbon feedstock for bioelectricity generation.
The formation of struvite crystals or magnesium ammonium phosphate (MgNH4PO4) in palm oil mill effluent (POME) occurs as early as in the secondary stage of POME treatment system. Its growth continues in the subsequent tertiary treatment which reduces piping diameter, thus affecting POME treatment efficiency. Hypothesis. The beneficial use of the crystal is the motivation. This occurrence is rarely reported in scientific articles despite being a common problem faced by palm oil millers. The aim of this study is to characterize struvite crystals found in an anaerobic digester of a POME treatment facility in terms of their physical and chemical aspects. The compositions, morphology and properties of these crystals were determined via energy dispersive spectroscopy (EDS), elemental analysis, scanning electron microscopy (SEM) and x-ray diffraction (XRD). Solubility tests were carried out to establish solubility curve for struvite from POME. Finally, crystal growth experiment was done applying reaction crystallization method to demonstrate struvite precipitation from POME. Results showed that high phosphorous (P) (24.85 wt%) and magnesium (Mg) (21.33 wt%) content was found in the struvite sample. Elemental analysis detected carbon (C), hydrogen (H), nitrogen (N) and sulfur (S) below 4 wt%. The crystals analysed by XRD in this study were confirmed as struvite with 94.8% struvite mineral detected from its total volume. Having an orthorhombic crystal system, struvite crystals from POME recorded an average density of 1.701 g cm-3. Solubility curve of struvite from POME was established with maximum solubility of 275.6 mg L-1 at pH 3 and temperature 40 °C. Minimum solubility of 123.6 mg L-1 was recorded at pH 7 and temperature 25 °C. Crystal growth experiment utilizing POME as the source medium managed to achieve 67% reduction in phosphorous content. This study concluded that there is a potential of harnessing valuable nutrients from POME in the form of struvite. Struvite precipitation technology can be adapted in the management of POME in order to achieve maximum utilization of the nutrients that are still abundant in POME. At the same time maximization of nutrient extractions from POME will also reduce pollutants loading in the final discharge.
A study of the kinetics and performance of solvent-yielding batch fermentation of individual sugars and their mixture derived from enzymic hydrolysis of sago starch by Clostridium acetobutylicum showed that the use of 30 g/L gelatinized sago starch as the sole carbon source produced 11.2 g/L total solvent, i.e. 1.5-2 times more than with pure maltose or glucose used as carbon sources. Enzymic pretreatment of gelatinized sago starch yielding maltose and glucose hydrolyzates prior to the fermentation did not improve solvent production as compared to direct fermentation of gelatinized sago starch. The solvent yield of direct gelatinized sago starch fermentation depended on the activity and stability of amylolytic enzymes produced during the fermentation. The pH optima for alpha-amylase and glucoamylase were found to be at 5.3 and 4.0-4.4, respectively. alpha-Amylase showed a broad pH stability profile, retaining more than 80% of its maximum activity at pH 3.0-8.0 after a 1-d incubation at 37 degrees C. Since C. acetobutylicum alpha-amylase has a high activity and stability at low pH, this strain can potentially be employed in a one-step direct solvent-yielding fermentation of sago starch. However, the C. acetobutylicum glucoamylase was only stable at pH 4-5, maintaining more than 90% of its maximum activity after a 1-d incubation at 37 degrees C.
In an attempt to reduce costs, the role of Bactec anaerobic blood culture in the detection of bacteraemia and fungaemia in children was evaluated. Results from 3167 sets of aerobic and anaerobic blood cultures from children admitted to the University Hospital, Kuala Lumpur during a one year period, were analysed. Four hundred and eight (12.9%) sets of blood cultures were positive, of which 348 sets (11.0%) from 201 patients were clinically significant. Of the 348 significant positive sets, organisms were isolated on 177 (50.9%) occasions from both aerobic and anaerobic bottles, on 136 (39.1%) occasions from the aerobic bottle only and 35 (10.0%) occasions from the anaerobic bottle only. No strict anaerobes were isolated, but clinically significant isolates recovered from the anaerobic bottle only included Klebsiella pneumoniae, Salmonella species, Enterobacter cloacae, Staphylococcus aureus, coagulase negative staphylococci, Streptococcus pneumoniae and Group B streptococcus. Patients with bacteraemia diagnosed solely by anaerobic culture were distributed evenly across the various paediatric subspecialities. When results from the anaerobic bottles were excluded, the overall isolation rate was reduced from 11% to 9.9%. Potential financial savings resulting from omission of anaerobic cultures must be balanced against the small number of bacteraemic episodes that could be missed. Undiagnosed bacteraemia may result in increased morbidity and mortality with its own attendant financial implications.
Isozymes of 23 cultures of the anaerobic rumen fungi and seven cultures of aerobic chytridiomycete fungi were analysed by PAGE. A total of 14 isozyme loci were successfully typed by PAGE. They were peptidase A & C-1, peptidase A & C-2, peptidase D-1, peptidase D-2, malate dehydrogenase-1, malate dehydrogenase-2, esterase-1, esterase-2, malic enzyme-1, malic enzyme-2, isocitrate dehydrogenase, shikimate dehydrogenase, phosphoglucomutase and 6-phosphogluconate dehydrogenase. Isozyme analysis can be used for studying the genetic relationships among the different anaerobic rumen fungi and the aerobic chytridiomycete fungi and the isozyme characteristics can serve as additional taxonomic criteria in the classification of the anaerobic rumen fungi. A dendrogram based on the isozyme data demonstrated that the anaerobic rumen fungi formed a cluster, indicating a monophyletic group, distinctly separated from the aerobic chytridiomycete fungi. Piromyces communis and P. minutus showed a close relationship but P. spiralis showed a more distant relationship to both P. communis and P. minutus. Piromyces as a whole was more related to Caecomyces than to Neocallimastix. Orpinomyces was also found to be more related to Piromyces and Caecomyces than to Neocallimastix. Orpinomyces intercalaris C 70 from cattle showed large genetic variation from O. joyonii, indicating that it is a different species.
The recalcitrant landfill leachate was anaerobically digested at various mixing ratios with labile synthetic wastewater to evaluate the degradation properties of recalcitrant wastewater. The proportion of leachate to the digestion system was increased in three equal steps, starting from 0% to 100%, and later decreased back to 0% with the same steps. The chemical oxygen demand (COD) for organic carbon and other components were calculated by analyzing the COD and dissolved organic carbon (DOC), and the removal efficiencies of COD carbon and COD others were evaluated separately. The degradation properties of COD carbon and COD others shifted owing to changing of substrate degradability, and the removal efficiencies of COD carbon and COD others were improved after supplying 100% recalcitrant wastewater. The UV absorptive property and total organic carbon (TOC) of each molecular size using high performance liquid chromatography (HPLC)-size exclusion chromatography (SEC) with UVA and TOC detectors were also investigated, and the degradability of different molecular sizes was determined. Although the SEC system detected extracellular polymeric substances (EPS), which are produced by microbes in stressful environments, during early stages of the experiment, EPS were not detected after feeding 100% recalcitrant wastewater. These results suggest that the microbes had acclimatized to the recalcitrant wastewater degradation. The high removal rates of both COD carbon and COD others were sustained when the proportion of labile wastewater in the substrate was 33%, indicating that the effective removal of recalcitrant COD might be controlled by changing the substrate's degradability.
This study involves the production of short-chain organic acids from kitchen wastes as intermediates for the production of biodegradable plastics. Flasks, without mixing were used for the anaerobic conversion of the organic fraction of kitchen wastes into short-chain organic acids. The influence of pH, temperature and addition of sludge cake on the rate of organic acids production and yield were evaluated. Fermentations were carried out in an incubator at different temperatures controlled at 30 degrees C. 40 degrees C, 50 degrees C, 60 degrees C and uncontrolled at room temperature. The pH was also varied at pH 5, 6, 7, and uncontrolled pH. 1.0 M phosphate buffer was used for pH control, and 1.0 M HCl and 1.0 M NaOH were added when necessary. Sludge cake addition enhanced the rate of maximum acids production from 4 days to 1 day. The organic acids produced were maximum at pH 7 and 50 degrees C i.e., 39.84 g/l on the fourth day of fermentation with a yield of 0.87 g/g soluble COD consumed, and 0.84 g/g TVS. The main organic acid produced was lactic acid (65-85%), with small amounts of acetic (10-30%), propionic (5-10%), and butyric (5-20%) acids. The results of this study showed that kitchen wastes could be fermented to high concentration of organic acids, which could be used as substrates for the production of biodegradable plastics.
Rubber latex effluent is a polluting source that has a high biochemical oxygen demand (BOD). It is estimated that about 100 million liters of effluent are discharged daily from rubber processing factories. Utilization of this effluent such as the use of a coupled system not only can reduce the cost of treatment but also yield a fermentation feedstock for the production of bioplastic. This study initially was carried out to increase the production of organic acids by anaerobic treatment of rubber latex effluent. It was found that through anaerobic treatment the concentration of organic acids did not increase. Consequently, separation of organic acids from rubber latex effluent by anion exchange resin was examined as a preliminary study of recovering acetic and propionic acids. However, the suspended solids (SS) content in the raw effluent was rather high which partially blocked the ion-exchange columns. Lime was used to remove the SS in the rubber latex effluent. After the lime precipitation process, organic acids were found to adsorb strongly onto the anion exchange resin. Less adsorption of organic acids onto the resin was observed before the lime precipitation. This was probably due to more sites being occupied by colloidal particles on the resin thus inhibiting the adsorption of organic acids. The initial concentration of organic acids in the raw effluent was 3.9 g/L. After ion exchange, the concentration of the organic acids increased to 27 g/L, which could be utilized for production of polyhydroxyalkanoates (PHA). For PHA accumulation stage, concentrated rubber latex effluent obtained from ion exchange resins and synthetic acetic acid were used as the carbon source. Quantitative analyses from fed batch culture via HPLC showed that the accumulation of PHA in Alcaligenes eutrophus was maximum with a concentration of 1.182 g/L when cultivated on synthetic acetic acid, corresponding to a yield of 87% based on its cell dry weight. The dry cell weight increased from 0.71 to 1.67 g/L. On the other hand, using concentrated rubber latex effluent containing acetic and propionic acids resulted in reduced PHA content by dry weight (14%) but the dry cell weight increased from 0.49 to 1.30 g/L. The results clearly indicated that the cells grow well in rubber latex effluent but no PHA was accumulated. This could be due to the high concentration of propionic acid in culture broth or other factors such as heavy metals. Thus further work is required before rubber latex effluent can be utilized as a substrate for PHA production industrially.
Isolates of anaerobic fungi obtained from the rumen, duodenum and faeces of sheep were identified as Piromyces mae based on their morphological characteristics observed using light microscopy. There was no significant morphological variation among the isolates of P. mae from the rumen, duodenum and faeces. Isozymes of 12 isolates of P. mae (one each from the rumen, duodenum and faeces from 4 different sheep) were analysed by PAGE. A total of 12 isozymes were studied and 5 isozyme loci were successfully typed. They were malic enzyme, malate dehydrogenase, shikimate dehydrogenase, alpha-esterase and beta-esterase. All the isolates of P. mae regardless of whether they were from the rumen, duodenum or faeces or from different animals produced very similar isozyme banding patterns for each of the enzyme systems. The similar isozyme profiles of the isolates indicate that they are of the same species although they exist in different regions of the alimentary tract.
Conventional aerobic and low-rate anaerobic processes such as pond and open-tank systems have been widely used in wastewater treatment. In order to improve treatment efficacy and to avoid greenhouse gas emissions, conventional treatment can be upgraded to a high performance anaerobic granular-sludge system. The anaerobic granular-sludge systems are designed to capture the biogas produced, rendering a potential for claims of carbon credits under the Kyoto Protocol for reducing emissions of greenhouse gases. Certified Emission Reductions (CERs) would be issued, which can be exchanged between businesses or bought and sold in international markets at the prevailing market prices. As the advanced anaerobic granular systems are capable of handling high organic loadings concomitant with high strength wastewater and short hydraulic retention time, they render more carbon credits than other conventional anaerobic systems. In addition to efficient waste degradation, the carbon credits can be used to generate revenue and to finance the project. This paper presents a scenario on emission avoidance based on a methane recovery and utilization project. An example analysis on emission reduction and an overview of the global emission market are also outlined.
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.