Cellulase production by Aspergillus fumigatus A4112 and the potential use of the enzyme in cooperation with surfactant to enhance floating oil recovery and methane production from palm oil mill effluent.

This research performed cellulase production by Aspergillus fumigatus A4112 and evaluated its potential use in palm oil mill effluent (POME) hydrolysis to recover oil simultaneously with the generation of fermentable sugar useful for biofuel production under non-sterilized conditions. Empty fruit bunch (EFB) without pretreatment was used as carbon source. The combination of nitrogen sources facilitated CMCase production. The maximum activity (3.27 U/mL) was obtained by 1.0 g/L peptone and 1.5 g/L (NH4)2SO4 and 20 g/L EFB at 40 °C for 7 days. High level of FPase activity (39.51 U/mL) was also obtained. Interestingly, the enzyme retained its cellulase activities more than 60% at ambient temperature over 15 days. In enzymatic hydrolysis, Triton X-100 was an effective surfactant to increase total oil recovery in the floating form. High yield of reducing sugar (50.13 g/L) and 21% (v/v) of floating oil was recoverable at 65 °C for 48 h. Methane content of the raw POME increased from 41.49 to 64.94% by using de-oiled POME hydrolysate which was higher than using the POME hydrolysate (59.82%). The results demonstrate the feasibility of the constructed process for oil recovery coupled with a subsequent step for methane yield enhancement in biogas production process that benefits the palm oil industry.

Antifungal and antiaflatoxigenic mechanism activity of freeze-dried culture filtrate of Streptomyces philanthi RL-1-178 on the two aflatoxigenic fungi and identification of its active component.

AIMS: The study reports the antifungal and antiaflatoxigenic mechanism activity of freeze-dried culture filtrate of Streptomyces philanthi RL-1-178 (DCF RL-1-178) against two aflatoxigenic strains (Aspergillus parasiticus and A. flavus) and identification of its active component. METHODS AND RESULTS: Significant inhibition in ergosterol biosynthesis by the DCF RL-1-178 appeared on the plasma membrane. Moreover, the DCF RL-1-178 showed dose-dependent inhibition of methylglyoxal (MG) (an aflatoxin inducer) biosynthesis and exhibited a novel antiaflatoxigenic action mechanism. Significant impairments in enzymatic [superoxide dismutase (SOD) and catalase (CAT)] and nonenzymatic [oxidized and reduced glutathione (GSH) and ratio of oxidized and reduced glutathione (GSSG)] anti-oxidative defense molecules were observed in the two aflatoxigenic cells. The active component of the DCF RL-1-178 was identified as natamycin. The natamycin exhibited against A. parasiticus and A. flavus with the minimum inhibitory concentration (MIC) values of 0.5 and 1.0 µg ml-1, respectively, while the minimum fungicidal concentration values were the same (4.0 µg ml-1). CONCLUSIONS: The DCF RL-1-178 containing natamycin exhibited the following effects: (1) inhibition of cellular ergosterol biosynthesis on plasma membrane, (2) reduction in MG (aflatoxin inducer) confirmed novel antiaflatoxigenic mechanism of action, and (3) caused remarkable debasement in antioxidant defense enzymes (SOD and CAT) and nonenzymatic defense molecules (GSH and GSSG) revealing biochemical mechanism of action.

Application of antifungal metabolites from Streptomyces philanthi RL-1-178 for maize grain coating formulations and their efficacy as biofungicide during storage.

The major safety risk of maize grain is contamination with mycotoxins. In this study, a maize-coating formulation containing freeze-dried culture filtrate of Streptomyces philanthi RL-1-178 (DCF RL-1-178) was developed and evaluated to prevent the growth of mycotoxins during maize grain storage. In vitro studies using confrontation tests on PDA plates indicated that S. philanthi RL-1-178 inhibited the growth of Aspergillus parasiticus TISTR 3276 (89.0%) and A. flavus PSRDC-4 (95.0%). The maize grain coating formulations containing the DCF RL-1-178 (0, 5, 10, and 15% (v/v)) and the polymer polyvinylpyrrolidone (PVP-K90, 4.0% (w/v)) were tested for their efficacy in In vitro and during 5 months storage. In In vitro assay, maize coating formular containing the optimum concentration (15.0%, v/v) of the DCF RL-1-178 exhibited 54.80% and 54.17% inhibition on the growth of A. parasiticus TISTR 3276 and A. flavus PSRDC-4 respectively. The inhibition was also illustrated by the microstructures of interactions between the coated maize grains with or without the DCF RL-1-178 and the fungal pathogens observed under microscope and SEM. Incorporating the DCF RL-1-178 or fungicidal Metalaxyl® into the polymer PVP-K90 maize grains coating resulted in the complete inhibition of the production of aflatoxin B1 (analysed by HPLC) by the two aflatoxigenic pathogens after 5 months storage at room temperature. However, the shelf-life was shortened to only 3 months during storage at room temperature with 90% relative humidity. Overall, the application of the 10-15% DCF RL-1-178 into the maize grain coating formular provides a new alternative measure to control the mycotoxins during storage for at least 5 months. The In vitro cell cytotoxicity study showed that a concentration of 15% (v/v) or 1000 µg/mL of the DCF RL-1-178 had a strong cytotoxic effect on Vero cells. These findings indicate that DCF RL-1-178 is a potential biofungicide for controlling mycotoxins contamination in maize seed storage for planting, but not maize grain storage for animal feed.

Utilization of palm oil mill effluent as a novel substrate for the production of antifungal compounds by Streptomyces philanthi RM-1-138 and evaluation of its efficacy in suppression of three strains of oil palm pathogen.

AIMS: This study aimed to use palm oil mill effluent (POME) as a renewable resource for the production of antifungal compounds by Streptomyces philanthi RM-1-138 against Ganoderma boninense, Ceratocystis paradoxa and Curvularia oryzae. METHODS AND RESULTS: The efficacy of antifungal compounds RM-1-138 against the three strains of fungal oil palm pathogen was evaluated both in vitro and on oil palm leaf segments. In vitro studies using confrontation tests on glucose yeast-malt extract (GYM) agar plates indicated that the strain RM-1-138 inhibited the growth of all three fungal pathogenic strains. The antifungal compounds produced in the GYM medium exhibited significantly higher inhibition (79%-100%) against the three fungal pathogens than using the diluted POME (50%) medium (80%-83% inhibition). The optimum condition for the production of antifungal compounds from the strain RM-1-138 was as following: POME of 47,966 mg L-1 chemical oxygen demand (COD), the initial pH at 7.0 and supplemented with yeast extract (0.4%). Meanwhile, severe morphological and internal abnormalities in C. oryzae hyphae were observed under a scanning electron microscope and transmission electron microscope. In vivo experiment on oil palm leaf segments indicated that the efficacy of the antifungal compounds RM-1-138 (DSI = 1.3) were not significantly difference in the suppression of Curvularia leaf spot compared with the two commercial chemical fungicides of mancozeb® (DSI = 1.0) and tetraconazole® (DSI = 1.3). CONCLUSIONS: Antifungal compounds produced by S. philanthi RM-1-138 grown in POME have the potential to inhibit fungal pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: The POME (about 47 mg L-1 COD) with the initial pH of 7.0 and supplementation of 0.4% nitrogen could be used as a culture medium for the growth and production of antifungal compounds of S. philanthi RL-1-138. In addition, the antifungal compound RM-1-138 could suppress the three strains of oil palm fungal pathogen tested on oil palm leaf segment.

Impact of environmental factors on Streptomyces spp. metabolites against Botrytis cinerea.

Botrytis cinerea is an economically important disease on numerous vegetables including tomato. From our previous studies, a spore suspension of Streptomyces philanthi RL-1-178 and RM-1-138 and Streptomyces mycarofaciens SS-2-243 showed strong inhibition against B. cinerea. In this study, the efficacy of their antifungal metabolites against B. cinerea was investigated after enhancing the production through the optimum culture medium and environmental conditions (temperature, light/dark cycle). In vitro studies indicated that glucose yeast-malt (GYM) agar and incubation at 28°C were optimal for growth and mass spore production of all three Streptomyces strains. Moreover, light/dark conditions had a positive effect on the growth and spore production of S. philanthi RM-1-138 and RL-1-178 but not on S. mycarofaciens SS-2-243. Both strains of S. philanthi possessed an antifungal activity against B. cinerea (100% inhibition) while S. mycarofaciens showed different results on PDA (83% inhibition) and GYM (88% inhibition) at the optimum incubation temperature at 21°C. The antifungal compounds from S. philanthi RM-1-138 exhibited the highest protection efficacy against B. cinerea on tomato leaves (82.89% and 0.33 cm2 lesion areas symptoms). The antifungal compounds RM-1-138, identified by GC-MS, were greatly altered based on components concentration under various temperatures and light/dark conditions. The anti-B. cinerea of S. philanthi RM-1-138 was established at a higher level in several metabolic compounds in the dark condition (11 and 32 antifungal compounds after incubation at 21°C and 28°C, respectively) than in the light condition (11 and 19 antifungal compounds after incubation at 21°C and 28°C, respectively). At 21°C, the dominant component was acetic acid (67.41% and 68.77% in light and dark conditions, respectively) while at 28°C, benzeneacetamide (43.58% in light) and propanamide (20.68% in the dark) were dominant. The results clearly demonstrated the significant influence of environmental factors on the production of antifungal metabolites of Streptomyces spp.

Efficacy of the antifungal metabolites of Streptomyces philanthi RL-1-178 on aflatoxin degradation with its application to prevent aflatoxigenic fungi in stored maize grains and identification of the bioactive compound.

Aflatoxin B1 is a potent carcinogen produced by Aspergillus flavus (A. flavus) and Aspergillus. parasiticus (A. parasiticus), mainly during grain storage. The efficacy of the freeze-dried culture filtrate of Streptomyces philanthi (S. philanthi) strain RL-1-178 (DCF) on degradation of aflatoxin B1 (AFB1) were evaluated and its bioactive compounds were identified. The DCF at a concentration of 9.0% (w/v) completely inhibited growth and AFB1 production of A. parasiticus TISTR 3276 and A. flavus PSRDC-4 after 7 days tested in yeast-extract sucrose (YES) medium and on stored maize grains after 28 and 14 days incubation, respectively. This indicated the more tolerance of A. parasiticus over A. flavus. The DCF and bacterial cells of S. philanthi were capable to degrade AFB1 by 85.0% and 100% for 72 h and 8 days, respectively. This confirmed the higher efficacy of the DCF over the cells. After separation of the DCF on thin-layer chromatography (TLC) plate by bioautography bioassay, each active band was identified by liquid chromatography-quadrupole time of flight mass spectrometer (LC-Q-TOF MS/MS). The results revealed two compounds which were identified as azithromycin and an unknown based on mass ions of both ESI+ and ESI- modes. The antifungal metabolites in the culture filtrate of S. philanthi were proved to degrade aflatoxin B1. It could be concluded that the DCF may be applied to prevent the growth of the two aflatoxin-producing fungi as well as the occurrence of aflatoxin in the stored maize grains.

The Degradation of Phenanthrene, Pyrene, and Fluoranthene and Its Conversion into Medium-Chain-Length Polyhydroxyalkanoate by Novel Polycyclic Aromatic Hydrocarbon-Degrading Bacteria.

Screening of high-efficient polycyclic aromatic hydrocarbon (PAH)-degrading bacteria is important due to environmental contamination by PAHs. In this study, sediment contaminated with phenanthrene (Phe), pyrene (Pyr), and fluoranthene (Fluo) was used as a source of bacteria. The ability of these isolated bacteria to convert PAHs into valuable products was determined. Based on a primary screening, 20 bacterial isolates were obtained; however, only three strains showed a good PAH-degrading ability, and were identified as Pseudomonas aeruginosa, Pseudomonas sp., and Ralstonia sp. PAH-degrading genes were detected in all isolates. Notably, all selected strains could degrade PAHs using the ortho or meta cleavage pathways due to the presence of catechol dioxygenase genes. The ability of isolated strains to convert PAHs into polyhydroxyalkanoate (PHA) was also evaluated in both single and mixed cultures. Single cultures of P. aeruginosa PAH-P02 showed 100% degradation of PAHs, with the highest biomass (1.27 ± 0.02 g l-1) and PHA content (38.20 ± 1.92% dry cell weight). However, degradative ability and PHA production were decreased when mixtures of PAHs were used. This study showed that P. aeruginosa, Pseudomonas sp., and Ralstonia sp. were able to degrade PAHs and convert them into medium-chain-length (mcl)-PHA. A high content of 3-hydroxydecanoate (3HD, C10) was observed in this study. The formation of mcl-PHA with high 3HD content from Pyr and Fluo, and the assessment of mixed cultures converting PAHs to mcl-PHA, were novel contributions.

Bacillus thermoamylovorans-Related Strain Isolated from High Temperature Sites as Potential Producers of Medium-Chain-Length Polyhydroxyalkanoate (mcl-PHA).

Thermotolerant bacteria producing medium-chain-length polyhydroxyalkanoate (mcl-PHA) were isolated from various thermal sites, including palm oil mill effluent, textile wastewater, and hot spring water, in Thailand. Fifteen strains were isolated at 45 °C using nutrient-rich (NR) medium. However, only six isolates produced mcl-PHA at 0.41 ± 0.01 g/L to 0.80 ± 0.01 g/L, representing a mcl-PHA content of 29.44% to 50.77% of the dry cell weight (DCW). The six strains of bacterial isolates could utilise a variety of substrates; all were identified as Bacillus thermoamylovorans. The highest mcl-PHA content (50.77% of the DCW) was accumulated by the B. thermoamylovorans strain PHA005 isolated from palm oil mill effluent. The mcl-PHA from strain PHA005 was composed of five different monomers, 3-hydroxyoctanoate (3HO), 3-hydroxydecanoate (3HD), 3-hydroxytetradecanoate (3HTD), 3-hydroxyhexadecanoic acid (3HHD), and 3-hydroxyoctadecanoic (3HOD), with a monomer content of 24.12, 15.50, 13.00, 39.25, and 8.13 mol%, respectively. The optimum temperature for B. thermoamylovorans strain PHA005 growth is 45 °C, and it can survive at up to 60 °C. This is a first report of PHA synthesis by a thermotolerant B. thermoamylovorans. Moreover, the high content of 3HHD monomers (39.25 mol%) has never been reported in Bacillus.

A rapid method for harvesting and immobilization of oleaginous microalgae using pellet-forming filamentous fungi and the application in phytoremediation of secondary effluent.

A rapid method for harvesting and immobilization of oleaginous microalgae using pellet-forming filamentous fungi was developed. The suitable conditions for pellet formation by filamentous fungi were determined. Among the strains tested, Trichoderma reesei QM 9414 showed superior pellet forming ability. Its pellets were used to harvest oleaginous microalga Scenedesmus sp. With increasing volume ratio of fungal pellets to microalgae culture up to 1:2, >94% of microalgal cells were rapidly harvested within 10 min. The ratio of fungal pellets could manipulate both harvesting time and initial concentration of microalgal cells in the pellets. The microalgae-fungal pellets were successfully used as immobilized cells for effective phytoremediation of secondary effluent from seafood processing plants under nonsterile condition. The chemical oxygen demand, total nitrogen, and total phosphorus removal were >74%, >44%, and >93%, respectively. The scanning electron microscopy showed that the microalgal cells were not only entrapped in the pellets but also got attached to the fungal hyphae with sticky exopolysaccharides, possibly secreted by the fungi. The extracted lipids from the pellets were mainly composed of C16-C18 (>83%) with their suitability as biodiesel feedstocks. This study has shown the promising strategy to rapidly harvest and immobilize microalgal cells and the possible application in phytoremediation of industrial effluent.

Inhibitory effects of acetophenone or phenylethyl alcohol as fumigant to protect soybean seeds against two aflatoxin-producing fungi.

The antifungal activity of acetophenone and phenylethyl alcohol prevents seed contamination by Aspergillus flavus TISTR 3041 and A. parasiticus TISTR 3276. Their effects on seed germination were investigated. In vitro results showed that 100 µL L-1 acetophenone completely inhibited (by 100%) growth, conidial germination, and sporulation of the two aflatoxin-producing fungi, while phenylethyl alcohol showed only weak inhibitory activity even at 1000 µL L-1. Exposure to acetophenone at 100 µL L-1 for 6 h could completely kill (100% death) both fungal strains, while phenylethyl alcohol showed much lower efficacy (53.12%). In vivo results revealed that fumigation with 100 µL L-1 acetophenone for 24 h completely controlled (100%) A. flavus TISTR 3041 on soybean seeds during a 14-day test but exhibited weak efficacy on A. parasiticus TISTR 3276 (31.77%). Phenylethyl alcohol (1000 µL L-1) demonstrated weak inhibitory effect against both strains. The two volatile compounds had no adverse effects on seed germination. SEM confirmed that acetophenone could completely inhibit conidia germination, and abnormal growth of both fungal strains was observed. Thus, acetophenone has high potential to protect soybean seeds against aflatoxin-producing fungi.

Factors affecting antifungal activity of Streptomyces philanthi RM-1-138 against Rhizoctonia solani.

Sheath blight disease of rice caused by Rhizoctonia solani Kühn is economically important disease in most of the world's rice growing areas. The disease causes severe yield losses of >20% of rice in Thailand. Our previous investigation reported the antifungal activity of Streptomyces philanthi RM-1-138 against R. solani PTRRC-9. In this study, glucose yeast-malt extract medium, initial pH of 7.5 and a temperature of 30 °C were found to be optimum for both cell growth and antifungal activity of S. philanthi RM-1-138. The inhibition of 94 and 100% on the growth of R. solani PTRRC-9 were achieved from the antifungal metabolites of the 6 and 9-days-old culture filtrates of S. philanthi RM-1-138, respectively. Heat treatment on the culture filtrate had slight effect on its antifungal activity. The culture broth demonstrated higher antifungal activity on growth of R. solani PTRRC-9 (90.4%) than the culture filtrate (31.5%) and its effective dose was at 0.1% (v/v). The present results indicated the possibilities of using either the culture broth or culture filtrate of S. philanthi RM-1-138 to inhibit growth of R. solani PTRRC-9.

Biosynthesis of 1,3-propanediol from recombinant E. coli by optimization process using pure and crude glycerol as a sole carbon source under two-phase fermentation system.

The environmental and nutritional condition for 1,3-propanediol (1,3-PD) production by the novel recombinant E. coli BP41Y3 expressing fusion protein were first optimized using conventional approach. The optimum environmental conditions were: initial pH at 8.0, incubation at 37 °C without shaking and agitation. Among ten nutrient variables, fumarate, (NH4)2HPO4 and peptone were selected to study on their interaction effect using the response surface methodology. The optimum medium contained modified Riesenberg medium (containing pure glycerol as a sole carbon source) supplemented with 63.65 mM fumarate, 3.80 g/L (NH4)2HPO4 and 1.12 g/L peptone, giving the maximum 1,3-PD production of 2.43 g/L. This was 3.5-fold higher than the original medium (0.7 g/L). Two-phase cultivation system was conducted and the effect of pH control (at 6.5, 7.0 and 8.0) was investigated under anaerobic condition by comparing with the no pH control condition. The cultivation system without pH control (initial pH of 8.0) gave the maximum values of 1.65 g/L 1,3-PD, the 1,3-PD production rate of 0.13 g/L h and the yield of 0.31 mol 1,3-PD/mol crude glycerol. Hence, using crude glycerol as a sole carbon source resulted in 32 % lower 1,3-PD production from this recombinant strain that may be due to the presence of various impurities in the crude glycerol of biodiesel plant. In addition, succinic acid was found to be a major product during fermentation by giving the maximum concentration of 11.92 g/L after 24 h anaerobic cultivation.

Potential for the integration of biological and chemical control of sheath blight disease caused by Rhizoctonia solani on rice.

Biological control using antagonistic microbes to minimize the use of chemical pesticides has recently become more prevalent. In an attempt to find an integrated control system for sheath blight, caused by Rhizoctonia solani in rice, Streptomyces philanthi RM-1-138, commercial formulations of Bacillus subtilis as Larminar® and B. subtilis strain NSRS 89-24+MK-007 as Biobest® and chemical fungicides including carbendazim®, validamycin®, propiconazole® and mancozeb® were applied alone and in combination with S. philanthi RM-1-138. In vitro experiments showed that all treatments tested did provide some control against mycelial growth and sclerotia production by R. solani PTRRS-9. In addition, the four chemical fungicides had no detrimental effects on S. philanthi RM-1-138 even at high concentrations (up to 100 µg/ml). The efficacy of S. philanthi RM-1-138, the commercial formulations of B. subtilis, chemical fungicides alone or in combination with S. philanthi RM-1-138 was also tested in a greenhouse experiment against sheath blight disease on rice plants. All treatments showed some protection of rice for sheath blight by 47-60 % when carbendazim® was applied alone and up to 74 % when combined with S. philanthi RM-1-138.

Exploring the potential of Crotalaria juncea flower extracts as a source of antioxidants, antimicrobials, and cytoprotective agents for biomedical applications.

Plants provide an unlimited source of bioactive compounds, possessing tremendous applications in the pharmaceutical industry. In the search for sources of antioxidants and antimicrobial agents against human pathogens, ethanol extracts of Crotalaria juncea flowers (CJ flower extract) were evaluated. The highest total phenolic (5.65 µg GAE/ml) and flavonoid (0.43 µg QE/ml) contents were observed in the 100 µg/ml CJ flower extract. To assess antioxidant activity, three in vitro antioxidant tests were employed: DPPH radical-scavenging, ABTS+ radical-scavenging, and hydroxyl radical-scavenging assay. The CJ flower extract demonstrated significant (P < 0.05) antioxidant activity, dependent on the percentage of solvent extraction and the specific assays utilized. The highest antioxidant activity was obtained with 100% ethanol extraction and using the hydroxyl radical-scavenging assay (56.63%). Antimicrobial activity was assessed against six human pathogens, including the fungi Microsporum gypseum and five Gram-positive bacteria (Propionibacterium acnes, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, and Streptococcus mutans), as well as one Gram-negative bacterium (Escherichia coli ). The CJ flower extract inhibited the growth of both fungal and bacterial pathogens. The cytotoxicity of the CJ flower extract was measured using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the highest concentration of the extract (100 µg/ml) did not affect L929 cell viability. Moreover, the CJ flower extract demonstrated the ability to suppress H2O2-induced toxicity in L929 cells. Overall, the CJ flower extract has potential as an alternative source for exploring new antioxidants, antimicrobial agents, and cytoprotectants that could prove valuable for biomedical applications.

Expression and applications of recombinant crustacean hyperglycemic hormone from eyestalks of white shrimp (Litopenaeus vannamei) against bacterial infection.

Crustacean hyperglycemic hormone (CHH) has many functions to regulate carbohydrate metabolism, ecdysis and reproduction including ion transport in crustaceans. The cDNA encoding CHH peptides containing 369 bp open reading frame encoding 122 amino acids was cloned from eyestalk of white shrimp (Litopenaeus vannamei) and was produced by a bacterial expression system. The biological activity of recombinant L. vannamei crustacean hyperglycemic hormone (rLV-CHH) was tested. The hemolymph glucose level of shrimp increased two-fold at 1h after the rLV-CHH injection and then returned to normal after 3h. In addition to the effect of rLV-CHH administration (25 µg/shrimp) on immunological responses of white shrimp against pathogenic bacteria, Vibrio harveyi was studied. Results showed that the blood parameters of shrimp injected with rLV-CHH; the THC, PO activity, serum protein level and clearance ability to V. harveyi, were also higher than those of Neg-protein and PBS-injected shrimp. The survival of shrimp injected with rLV-CHH was significantly higher (66.0%) than shrimp that injected with Neg-protein (33.3%) and PBS (28.9%) after 14 days. It is possible that the administration of rLV-CHH in L. vannamei exhibited a higher immune response related to resistance against V. harveyi infection.

Improvement of empty palm fruit bunches biodegradability and biogas production by integrating the straw mushroom cultivation as a pretreatment in the solid-state anaerobic digestion.

Empty fruit bunches (EFB) have low biodegradability and restrict their commercial utilization in biogas plants. Integration of straw mushroom (Volvariella volvacea) cultivation as a function of bio-pretreatment on EFB to improve biodegradability and methane production by solid-state anaerobic digestion (SS-AD) was investigated. The mushroom yield was 47.3 kg·tonne-1 EFB with remaining weight in spent mushroom-EFB (S-mEFB) of 82%. The cellulose, hemicellulose, and lignin of EFB were degraded by 3.3%, 21.3%, and 17.6%, respectively, with an increased surface area of S-mEFB. The biodegradability of S-mEFB (62.7%) was 2 times higher than raw EFB (33.5%) with the highest methane yield and production of 281 mL CH4·g-1 VS and 50.6 m3·tonne-1 S-mEFB, respectively. The co-digestion of S-mEFB with 5% v/w POME had highest methane yield of 405 mL CH4·g-1 VS with biodegradability of 90.8%. Integrating straw mushroom cultivation with SS-AD is a promising strategy for achieving an environmentally friendly and economically feasible process.

Strategies for recovery of imbalanced full-scale biogas reactor feeding with palm oil mill effluent.

BACKGROUND: Full-scale biogas production from palm oil mill effluent (POME) was inhibited by low pH and highly volatile fatty acid (VFA) accumulation. Three strategies were investigated for recovering the anaerobic digestion (AD) imbalance on biogas production, namely the dilution method (tap water vs. biogas effluent), pH adjustment method (NaOH, NaHCO3, Ca(OH)2, oil palm ash), and bioaugmentation (active methane-producing sludge) method. The highly economical and feasible method was selected and validated in a full-scale application. RESULTS: The inhibited sludge from a full-scale biogas reactor could be recovered within 30-36 days by employing various strategies. Dilution of the inhibited sludge with biogas effluent at a ratio of 8:2, pH adjustment with 0.14% w/v NaOH, and 8.0% w/v oil palm ash were considered to be more economically feasible than other strategies tested (dilution with tap water, or pH adjustment with 0.50% w/v Ca(OH)2, or 1.25% NaHCO3 and bioaugmentation) with a recovery time of 30-36 days. The recovered biogas reactor exhibited a 35-83% higher methane yield than self-recovery, with a significantly increased hydrolysis constant (kH) and specific methanogenic activity (SMA). The population of Clostridium sp., Bacillus sp., and Methanosarcina sp. increased in the recovered sludge. The imbalanced full-scale hybrid cover lagoon reactor was recovered within 15 days by dilution with biogas effluent at a ratio of 8:2 and a better result than the lab-scale test (36 days). CONCLUSION: Dilution of the inhibited sludge with biogas effluent could recover the imbalance of the full-scale POME-biogas reactor with economically feasible and high biogas production performance.

Phototropic H2 production by a newly isolated strain of Rhodopseudomonas palustris.

Rhodopseudomonas palustris TN1 was isolated from Songkhla Lake, Thailand. It phototrophically generates H(2) from the predominant volatile fatty acids (VFAs) produced from microbial dark-fermentations of palm oil milling effluent; yields from 20 mM butyrate, acetate and propionate were 4.7, 2.5, and 1.7 mol H(2) mol VFA(-1) with light efficiencies of 1.8, 1, and 0.2%, respectively. Optimum conditions were pH 7 and 3000 lux, although production was reduced by only 33% at 1000 lux. CO(2) evolution never exceeded 9 mmol mol VFA(-1).

Comparative assessment of single-stage and two-stage anaerobic digestion for biogas production from high moisture municipal solid waste.

BACKGROUND: Anaerobic digestion (AD) is a suitable process for treating high moisture MSW with biogas and biofertilizer production. However, the low stability of AD performance and low methane production results from high moisture MSW due to the fast acidify of carbohydrate fermentation. The effects of organic loading and incineration fly ash addition as a pH adjustment on methane production from high moisture MSW in the single-stage AD and two-stage AD processes were investigated. RESULTS: Suitable initial organic loading of the single-stage AD process was 17 gVS L-1 at incineration fly ash (IFA) addition of 0.5% with methane yield of 287 mL CH4 g-1 VS. Suitable initial organic loading of the two-stage AD process was 43 gVS L-1 at IFA addition of 1% with hydrogen and methane yield of 47.4 ml H2 g-1 VS and 363 mL CH4 g-1 VS, respectively. The highest hydrogen and methane production of 8.7 m3 H2 ton-1 of high moisture MSW and 66.6 m3 CH4 ton-1 of high moisture MSW was achieved at organic loading of 43 gVS L-1 at IFA addition of 1% by two-stage AD process. Biogas production by the two-stage AD process enabled 18.5% higher energy recovery than single-stage AD. The 1% addition of IFA into high moisture MSW was useful for controlling pH of the two-stage AD process with enhanced biogas production between 87-92% when compared to without IFA addition. Electricity production and energy recovery from MSW using the coupled incineration with biogas production by two-stage AD process were 9,874 MJ ton-1 MSW and 89%, respectively. CONCLUSIONS: The two-stage AD process with IFA addition for pH adjustment could improve biogas production from high moisture MSW, as well as reduce lag phase and enhance biodegradability efficiency. The coupled incineration process with biogas production using the two-stage AD process was suitable for the management of MSW with low area requirement, low greenhouse gas emissions, and high energy recovery.

Influence of precursors and inhibitor on the production of extracellular 5-aminolevulinic acid and biomass by Rhodopseudomonas palustris KG31.

5-Aminolevulinic acid (ALA) and the biomass of photosynthetic bacteria, Rhodopseudomonas palustris KG31, have very high potential for development and exploitation as bioherbicide and biofertilizer respectively. In this work, the effects of two precursors and an inhibitor of aminolevulinic dehydratase (ALAD) added to the VFA culture medium on the production of ALA and biomass were investigated. The experimental runs were carried out according to a Box-Behnken design. The precursors were added to the medium at the beginning of cultivation, while the inhibitor was added after 24 h. Statistical analysis indicated that levulinic acid (LA) has a positive effect on ALA production while glycine has a negative effect on biomass production. In order to enhance both ALA and biomass products, the most suitable medium was VFA medium supplemented with 3.0 mM glycine and 10 mM LA, giving ALA and biomass of 182.91 microM and 3.1 gDCW/l within 54 h.