Methane potential and biodegradability of rice straw, rice husk and rice residues from the drying process

Agricultural solid residues are a potential renewable energy source. Rice harvesting and production in Sancti Spíritus province, Cuba, currently generates residues without an environmentally sustainable disposal route. Rice residues (rice straw, rice husk and rice residues from the drying process) are potentially an important carbon source for anaerobic digestion. For this paper, rice residues were placed for 36 days retention time in anaerobic batch reactor environments at both mesophilic (37 °C) and thermophilic (55 °C) conditions. Biogas and methane yield were determined as well as biogas composition. The results showed that rice straw as well as rice residues from the drying process had the highest biogas and methane yield. Temperature played an important role in determining both biogas yield and kinetics. In all cases, rice straw produced the highest yields; under mesophilic conditions the biogas yield was 0.43 m(3) kg(VS)(-1), under thermophilic conditions biogas yield reached 0.52 m(3) kg(VS)(-1). In the case of the rice husk, the biodegradability was very low. Methane content in all batches was kept above 55% vol. All digested material had a high carbon:nitrogen (C:N) ratio, even though significant biodegradation was recorded with the exception of rice husk. A first-order model can be used to describe the rice crop residues fermentation effectively.     

Ammonia, a selective agent for methane production by syntrophic acetate oxidation at mesophilic temperature.

In biogas processes, methane production from acetate proceeds by either aceticlastic methanogenesis or through syntrophic acetate oxidation (SAO). In the present study, the pathway for methane production from acetate was analysed; i) during a gradual increase of the ammonia concentration (final concentration 7 g NH(4)(+) -N/L) in a semi-continuous lab-scale anaerobic digester (4.3 L), operating at mesophilic temperature (37 degrees C) or ii) in diluted enrichment cultures (100 ml) experiencing a gradual increase in ammonia, sodium, potassium and propionic acid. The pathway for methane formation was determined by calculating the (14)CO(2)/(14)CH(4) ratio after incubating samples with (14)C-2-acetate. In the anaerobic digester, as well as in the enrichment cultures, the (14)CO(2)/(14)CH4 ratio clearly increased with increasing ammonium-nitrogen concentration, i.e. as the ammonia concentration increased, a shift from the aceticlastic mechanism to the syntrophic pathway occurred. The shift was very distinct and occurred as the NH(4)(+) -N concentration rose above 3 g/l. No shift in pathway was seen during increasing concentrations of sodium, potassium or propionic acid. The shift to SAO in the biogas digester resulted in a twofold decrease in the specific gas and methane yield.     

Intracellular biopolymer productions using mixed microbial cultures from fermented POME.

This study aimed to produce polyhydroxyalkanoates (PHAs) from organic wastes by mixed bacterial cultures using anaerobic-aerobic fermentation systems. Palm oil mill effluent (POME) was used as an organic source, which was cultivated in a two-step-process of acidogenesis and acid polymerization. POME was operated in a continuous flow anaerobic reactor to access volatile fatty acids (VFAs) for PHAs production. During fermentation, VFA concentration was produced in the range of 5 to 8 g/L and the COD concentration reduced up to 80% from 65 g/L. The VFA from anaerobic fermentation was then utilised for PHA production using a mixed culture in availability of aerobic bioreactor. Production of PHAs was recorded high when using a high volume of substrates because of the higher VFA concentration. Even though the maximum PHA content was observed at only 40% of the cell dried weight (CDW), their production and performance are significant in mixed microbial culture.     

Degradation characteristics of polylactide in thermophilic anaerobic digestion with hyperthermophilic solubilization condition

To test whether hyperthermophilic treatment promotes polylactide (PLA) dissolution and methane conversion under anaerobic digestion conditions, a single thermophilic control reactor (55 °C) and a two-phase system consisting of a hyperthermophilic reactor (80 °C) and a thermophilic reactor (55 °C) were continuously fed with a mixture of PLA and artificial kitchen garbage. In Runs 1 and 2, the PLA dissolution ratios in the two-phase system were 79.2 ± 6.5% and 85.2 ± 7.0%, respectively, higher than those of the control. Batch experimental results indicated that hyperthermophilic treatment could promote PLA dissolution to a greater degree as compared with single thermophilic treatment and that ammonia addition also had a promotional effect on PLA dissolution. In the two-phase system, after hyperthermophilic treatment, dissolved PLA was converted to methane gas under the subsequent thermophilic condition.     

Maize mono-digestion efficiency: results from laboratory tests

A laboratory experimental campaign was carried out in order to assess the optimal configuration for the anaerobic digestion of a mixture of sweet corn and ensiled maize. Batch hydrolysis tests were conducted at 35 and 55 °C and at four different particle sizes (2, 5, 20 and 50 mm) obtained by manual chopping and sieving. Chemical pre-treatment by 24 h incubation at various acid and alkaline pH was also considered for its potential to increase the maize methane yield. Results suggest that the hydrolytic phase proceeds significantly faster under thermophilic conditions. Significant differences in the solubilization rate were also observed when comparing coarse (20-50 mm) with fine (2-5 mm) particles, while 2 and 5 mm particles were solubilized at similar rates. No advantages from the chemical pre-treatment, in terms of solubilization efficiency and biomethanization potential were observed. According to these preliminary results, a two-stage semi-continuous laboratory plant consisting of a thermophilic hydrolytic reactor followed by a mesophilic methanogenic reactor was operated for 110 days. Steady state loading parameters were: influent concentration (maize mixture diluted in tap water) of 46 g VS/L, hydraulic retention time of 31 d, organic loading rate of 1.5 g VS/L/d. Alkalinity was dosed to the methanogenic reactor to avoid pH drops. Collected data allowed the average biodegradation efficiency to be estimated at around 60-65%.     

Syntrophic acetate oxidation in two-phase (acid-methane) anaerobic digesters

The microbial processes involved in two-phase anaerobic digestion were investigated by operating a laboratory-scale acid-phase (AP) reactor and analyzing two full-scale, two-phase anaerobic digesters operated under mesophilic (35 °C) conditions. The digesters received a blend of primary sludge and waste activated sludge (WAS). Methane levels of 20% in the laboratory-scale reactor indicated the presence of methanogenic activity in the AP. A phylogenetic analysis of an archaeal 16S rRNA gene clone library of one of the full-scale AP digesters showed that 82% and 5% of the clones were affiliated with the orders Methanobacteriales and Methanosarcinales, respectively. These results indicate that substantial levels of aceticlastic methanogens (order Methanosarcinales) were not maintained at the low solids retention times and acidic conditions (pH 5.2-5.5) of the AP, and that methanogenesis was carried out by hydrogen-utilizing methanogens of the order Methanobacteriales. Approximately 43, 31, and 9% of the archaeal clones from the methanogenic phase (MP) digester were affiliated with the orders Methanosarcinales, Methanomicrobiales, and Methanobacteriales, respectively. A phylogenetic analysis of a bacterial 16S rRNA gene clone library suggested the presence of acetate-oxidizing bacteria (close relatives of Thermacetogenium phaeum, 'Syntrophaceticus schinkii,' and Clostridium ultunense). The high abundance of hydrogen consuming methanogens and the presence of known acetate-oxidizing bacteria suggest that acetate utilization by acetate oxidizing bacteria in syntrophic interaction with hydrogen-utilizing methanogens was an important pathway in the second-stage of the two-phase digestion, which was operated at high ammonium-N concentrations (1.0 and 1.4 g/L). A modified version of the IWA Anaerobic Digestion Model No. 1 (ADM1) with extensions for syntrophic acetate oxidation and weak-acid inhibition adequately described the dynamic profiles of volatile acid production/degradation and methane generation observed in the laboratory-scale AP reactor. The model was validated with historical data from the full-scale digesters.     

Anaerobic pond treatment of wastewater containing sulphate.

Anaerobic ponds are usually used for treatment of industrial and agricultural wastes which contain high organic matter and sulphate. Competition for substrate between sulphate reducing bacteria and methane producing archaea, and the inhibitory effects of sulphide produced from microbial sulphate reduction reported in the literature varied considerably. In this research, a laboratory scale column-in-series anaerobic pond reactor, consisting of five cylindrical columns of acrylic tubes, was operated to evaluate the effect of COD and sulphate ratio on pond performance treating wastewater containing high organic matter and sulphate from a tapioca starch industry. The result depicted that no adverse effect of COD:SO4 ratios between 5 and 20 on overall COD removal performance of anaerobic pond operated with organic loading rate (OLR) of 150 to 600 g COD/m3d. Sulphate reducing bacteria could out-compete methane producing archaea for the same substrate at COD:SO4 ratio equal to or lower than 5 and OLR greater than 300 g COD/m3d. Sulphide inhibition was not observed on overall performance of pond up to an influent sulphate concentration of 650 mg/L.     

Methanotrophic biodegradation of cis-1,2-dichloroethylene in a continuously fed fixed-film bioreactor.

Co-metabolic biodegradation of cis-dichloroethylene (cis-DCE) was investigated in a bench-scale fixed-film bioreactor inoculated with a mixed culture of methane oxidising bacteria. The aim of this work was to identify factors that affect the cis-DCE biodegradation. It was observed that the presence of methane was necessary to enhance the biodegradation of cis-DCE, but an excess of methane inhibited the cis-DCE removal. cis-DCE did not inhibit the methane biodegradation at concentrations up to 300 microg/L. Maximum cis-DCE removal was observed with a methane bulk concentration ranging from 0.2 to 0.7 mg/L. It was found that the activity of the biofilm was located in the upper 100 microm of the biofilm. On the basis of this study it is concluded that careful control of the oxygen and methane concentrations as well as of the biofilm thickness is necessary in order to optimise the biodegradation of cis-DCE in fixed film bioreactors.     

Anaerobic biodegradability of Tween surfactants used as a carbon source for the microbial reductive dechlorination of hexachlorobenzene.

Three structurally-related, nonionic, polysorbate surfactants (Tween 60, 61, and 65) were used as the sole carbon source to sustain the microbial, sequential reductive dechlorination of hexachlorobenzene (HCB) in a mixed, methanogenic culture derived from a contaminated estuarine sediment. The surfactants were partially degraded and fermented to methane with no measurable accumulation of volatile fatty acids, indicating that methanogenesis was rapid relative to the rates of hydrolysis and acidogenesis. Addition of the methanogenesis inhibitor 2-bromoethanesulfonic acid resulted in acetate accumulation without impact on the sequential dechlorination of HCB. An anaerobic biodegradability assay was performed and the following data were obtained for the Tween 60, 61, and 65, respectively: 53, 62, and 62% COD destruction; 35, 57, and 48% COD to methane conversion; and 38, 38, and 45% COD to acetate conversion. These data suggest that the hydrophobic moiety (stearate) of the surfactants was preferentially degraded, most likely through beta-oxidation, to acetate and ultimately to methane and carbon dioxide. Between 38 and 47% of the initial surfactant COD remained after 46 d incubation, which most likely corresponds to the hydrophilic polyoxyethylene moiety. An anaerobic biodegradation pathway of the Tween surfactants is proposed.     

Detection of fatty acid beta-oxidizing syntrophic bacteria by fluorescence in situ hybridization.

A new 16S rRNA-targeted oligonucleotide probe, specific for the cluster of fatty acid beta-oxidizing syntrophic bacteria of the family Syntrophomonadaceae was designed for fluorescence in situ hybridization. This probe was evaluated with target as well as non-target cultures. Moreover this probe was assessed with butyrate and oleate degrading enrichment cultures and methanogenic sludges from full-scale plants. The results showed that the probe revealed the presence of fatty acid beta-oxidizing syntrophic bacteria in some of the samples analyzed. However, cell quantification was possible only in enrichment cultures and in a flocculent sludge from a reactor that treats lipid-rich wastewaters, but not in methanogenic granular sludges from upflow anaerobic sludge blanket reactors.     

First data on lipids and microorganisms of deepwater endemic sponge Baikalospongia intermedia and sediments from hydrothermal discharge area of the Frolikha Bay (North Baikal,Siberia)

In this report, we study lipid components (fatty acids with various degrees of unsaturation, aldehydes and sterols) of deep-water sponges Baikalospongia intermedia and sediments sampled using deep-water manned submersibles “Mir” at a hydrothermal vent of Frolikha Bay at depths of 400–450 m. It was found that unsaturated fatty acids predominate in the FA-composition of the sponges: very long chain demospongic acid 26:3, as well as monounsaturated 24:1 and isomers of acids 16:1 and 18:1. Among the saturated fatty acids in addition to the stearic 18:0, palmitic 16:0 and heptadecanoic 17:0 acids, significant amounts of acids of microbial origin (iso-palmitic i16:0, isostearic i18:0, oxy-stearic 10 h18) were detected. Using the method of mass spectrometry of microbial markers, we conclude, that the main microbial components of sponges B. intermedia and sediments from Frolikha Bay are methanotrophic microorganisms.     

Monitoring biodegradation of diesel fuel in bioventing processes using in situ respiration rate.

An in situ measuring system of respiration rate was applied for monitoring biodegradation of diesel fuel in a bioventing process for bioremediation of diesel contaminated soil. Two laboratory-scale soil columns were packed with 5 kg of soil that was artificially contaminated by diesel fuel as final TPH (total petroleum hydrocarbon) concentration of 8,000 mg/kg soil. Nutrient was added to make a relative concentration of C:N:P = 100:10:1. One soil column was operated with continuous venting mode, and the other one with intermittent (6 h venting/6 h rest) venting mode. On-line O2 and CO2 gas measuring system was applied to measure O2 utilisation and CO2 production during biodegradation of diesel for 5 months. Biodegradation rate of TPH was calculated from respiration rate measured by the on-line gas measuring system. There were no apparent differences between calculated biodegradation rates from two columns with different venting modes. The variation of biodegradation rates corresponded well with trend of the remaining TPH concentrations comparing other biodegradation indicators, such as C17/pristane and C18/phytane ratio, dehydrogenase activity, and the ratio of hydrocarbon utilising bacteria to total heterotrophic bacteria. These results suggested that the on-line measuring system of respiration rate would be applied to monitoring biodegradation rate and to determine the potential applicability of bioventing process for bioremediation of oil contaminated soil.     

Calibration of a simple model for waste stabilisation pond performance in seasonal climates

The paper describes the calibration of a model for waste stabilisation pond (WSP) performance in seasonal climates, based on the use of readily available climate data sets. Calibration data were taken from a wide geographical area of Canada and the USA, including coastal and moderately seasonal sites. Good agreement with measured values was shown using a biochemical oxygen demand (BOD) decay constant of 0.3 day(-1) for facultative ponds and 0.07-0.1 day(-1) for storage/maturation ponds with a temperature-related Arrhenius constant of 1.05, and a fixed BOD decay constant of 0.007 day(-1) at water temperatures below 0 °C. The results suggested that such models could potentially be used as the basis for WSP design guidelines tailored to a wide range of climatic conditions.     

Co-digestion of manure and biowaste according to the EC animal by-products regulation and Finnish national regulations.

The objective of this study was to compare methane production and characteristics of digested material in anaerobic digestion concepts according to the Animal By-Products Regulation (ABP-Regulation) of the EC (hygienisation of biowaste for 1 hour at 70 degrees C, particle size < 12 mm) and Finnish national regulations (treatment temperature 55 degrees C, feeding interval 24 h, hydraulic retention time (HRT) 20 d, particle size < 40 mm) and with small variations in treatment methods for treating manure and biowaste. Moreover, the survival of three different salmonella bacteria in these processes was studied. Hygienisation of biowaste prior to digestion at 35 degrees C enhanced methane production by 14-18% compared to similar treatment without hygienisation. The differences in treatment temperature, HRT and hygienisation of biowaste prior to digestion did not significantly affect the characteristics of digested material. The concepts according to the ABP-Regulation and Finnish national regulations were effective in destroying salmonella bacteria to an undetectable level.     

Inhibitory effects of nitrate reduction on methanogenesis in the presence of different electron donors.

The preferential utilization of different electron donors and their effects on the nitrate reduction and methanogenesis in a mixed, mesophilic (35 degrees C) methanogenic culture were investigated. Batch methanogenic cultures were fed with dextrin/peptone (D/P), propionate, acetate, and H(2)/CO(2) at an initial COD of 500 mg/L and an initial nitrate concentration of 50 mg N/L. Immediate cessation of methane production was observed in all nitrate-amended cultures. Methane production completely recovered in the D/P- and acetate-fed cultures, and partially recovered or did not recover in the propionate- and H(2)/CO(2)-fed, nitrate-amended cultures, respectively. Accumulation of denitrification intermediates was observed in both the propionate- and H(2)/CO(2)-fed cultures, which resulted in inhibition of fermentation and/or methanogenesis. The fastest and the slowest nitrate reduction were observed in the acetate- and propionate-fed cultures, respectively.     

Methane production and solids destruction in an anaerobic solid waste reactor due to post-reactor caustic and heat treatment.

This study was undertaken to determine the feasibility of caustic and heat treatment of sludge from a dry anaerobic reactor (DAR) with respect to increased methane production and solids destruction. The DAR was operated semi-continuously at 55 degrees C on sized-reduced municipal solid waste at a solids retention time of 15 days. A respirometer was employed to monitor the extent and rate of methane production from anaerobic biodegradation of DAR sludge that was treated with caustic and heat. Results indicate that caustic and heat treatment at 50 degrees C and 175 degrees C increased methane production by 22% and 52%, respectively. Also, volatile solids destruction increased from 46% to 58% and 83%, respectively. Based on these results, economic analysis for a full-scale 10(5) kg/d facility suggests that annual project revenue for 50 degrees C and 175 degrees C treatment is estimated at $21,000 and $445,000, respectively.     

Anaerobic digestion of corn ethanol thin stillage in batch and by high-rate down-flow fixed film reactors

Thin stillage (CTS) from a dry-grind corn ethanol plant was evaluated as a carbon source for anaerobic digestion (AD) by batch and high rate semi-continuous down-flow stationary fixed film (DSFF) reactors. Biochemical methane potential (BMP) assays were carried out with CTS concentrations ranging from approximately 2,460-27,172 mg total chemical oxygen demand (TCOD) per litre, achieved by diluting CTS with clean water or a combination of clean water and treated effluent. High TCOD, SCOD and volatile solids (VS) removal efficiencies of 85 ± 2, 94 ± 0 and 82 ± 1% were achieved for CTS diluted with only clean water at an organic concentration of 21,177 mg TCOD per litre, with a methane yield of 0.30 L methane per gram TCOD(removed) at standard temperature and pressure (STP, 0 °C and 1 atmosphere). Batch studies investigating the use of treated effluent for dilution showed promising results. Continuous studies employed two mesophilic DSFF anaerobic digesters treating thin stillage, operated at hydraulic retention times (HRT) of 20, 14.3, 8.7, 6.3, 5 and 4.2 d. Successful digestion was achieved up to an organic loading rate (OLR) of approximately 7.4 g TCOD L(-1)d(-1) at a 5 d HRT with a yield of 2.05 LCH(4) L(-1)d(-1) (at STP) and TCOD and VS removal efficiencies of 89 ± 3 and 85 ± 3%, respectively.     

Anaerobic microbial LCFA degradation in bioreactors

This paper reviews recent results obtained on long-chain fatty acids (LCFA) anaerobic degradation. Two LCFA were used as model substrates: oleate, a mono-unsaturated LCFA, and palmitate, a saturated LCFA, both abundant in LCFA-rich wastewaters. 16S rRNA gene analysis of sludge samples submitted to continuous oleate- and palmitate-feeding followed by batch degradation of the accumulated LCFA demonstrated that bacterial communities were dominated by members of the Clostridiaceae and Syntrophomonadaceae families. Archaeal populations were mainly comprised of hydrogen-consuming microorganisms belonging to the genus Methanobacterium, and acetate-utilizers from the genera Methanosaeta and Methanosarcina. Enrichment cultures growing on oleate and palmitate, in the absence or presence of sulfate, gave more insight into the major players involved in the degradation of unsaturated and saturated LCFA. Syntrophomonas-related species were identified as predominant microorganisms in all the enrichment cultures. Microorganisms clustering within the family Syntrophobacteraceae were identified in the methanogenic and sulfate-reducing enrichments growing on palmitate. Distinct bacterial consortia were developed in oleate and palmitate enrichments, and observed differences might be related to the different degrees of saturation of these two LCFA. A new obligately syntrophic bacterium, Syntrophomonas zehnderi, was isolated from an oleate-degrading culture and its presence in oleate-degrading sludges detected by 16S rRNA gene cloning and sequencing.     

Evaluation of a two-stage hydrothermal process for enhancing acetic acid production using municipal biosolids

A two-stage hydrothermal process aimed at improving acetic acid production using municipal biosolids was evaluated against thermal hydrolysis and conventional wet oxidation process in a 600 ml Parr batch reactor. Thermal hydrolysis was conducted at 140 °C, wet oxidation at 220 °C and the two-stage process, which acted as a series combination of thermal hydrolysis and wet oxidation, at 220 °C. Initial pressure of 1 MPa was maintained in all the three processes. The results indicated that the highest acetic acid production of up to 58 mg/g dry solids feed was achieved in the wet oxidation process followed by the two-stage process with 36 mg/g dry solids feed and 1.8 mg/g dry solids feed for thermal hydrolysis. The acetic acid yield obtained by the thermal processes increased from 0.4% in the thermal hydrolysis process to 12% during the single stage wet oxidation, with the two-stage process achieving 8%. The purity of the acetic acid improved from 1% in thermal hydrolysis to 38% in the wet oxidation process. The two-stage process achieved acetic acid purity of 25%. This work demonstrated no enhancement of acetic acid production by the two-stage concept compared with the single stage wet oxidation process. This is in contrast to similar work by other researchers, investigated on carbohydrate biomass and vegetable wastes using hydrogen peroxide as the oxidant. However, the data obtained revealed that substrate specificity, reaction severity or oxidant type is clearly important in promoting reaction mechanisms which support enhanced acetic acid production using municipal biosolids.     

CFD (computational fluid dynamics) modelling of baffles for optimizing tropical waste stabilization pond systems.

CFD modelling of the incorporation of two baffles equally spaced along the longitudinal axis of the pond and with a length equal to 70% of the pond breadth, indicated a potential improvement in the removal of E. coli in a 4-day secondary facultative pond at 25degrees C from 5 x 10(6) per 100 ml in the effluent from a 1-day anaerobic pond to 4 x 10(4) per 100 ml; the reduction in an un-baffled pond was an order of magnitude less effective. The addition of a similarly baffled 4-day primary maturation pond reduced the effluent E. coli count to 340 per 100 ml; the reduction in an un-baffled series was two orders of magnitude less effective. Well designed baffles thus have considerable potential for reducing pond area requirements and hence costs in the hot tropics. These very promising results highlight the need for field studies on baffled pond systems to validate (or allow calibration) of the CFD model used in this study.