Potential for using enriched cultures and thermotolerant bacterial isolates for production of biohydrogen from oil palm sap and microbial community analysis

A limited number of bacteria can convert oil palm (Elaeis guineensis) sap to hydrogen with satisfactory yield and productivity. In this study, a total of 18 fermentative enriched cultures and 36 newly isolated thermotolerant bacterial strains were compared for hydrogen production from oil palm (OP) sap. The new isolates were obtained from hot springs, palm oil mill effluent and oil palm sap. The test was conducted in three steps: (i) a test for hydrogen production from mixed substrates (cellulose, starch, xylose, and glucose) and OP sap; (ii) a test for substrate concentration tolerance; and (iii) a test for thermotolerance. Five enriched candidates for each of the hydrogen producers were selected according to the criteria defined for the screening test. The hydrogen production of these selected bacterial strains from hot springs were cultivated in batch fermentation of oil palm sap at room temperature (30 ± 2 °C). Five enriched cultures, namely 81RN1, OPS, 85RN5, 89SR3-2 and 112YL1 were found to give high cumulative hydrogen formation of 1085, 1009, 994, 983 and 778 mL H₂/L-OP sap, respectively, with the hydrogen content of 29.8, 29.4, 28.7, 27.1 and 27.5%, respectively. PCR–DGGE profiling showed that all these five enriched cultures consisted of species closely related to the genus Clostridium sp. based on the 16S rRNA gene. For pure cultures, the top five hydrogen producers were the isolates encoded as PS-3, PS-4, PS-5, PS-7 and PS-8 exhibiting the hydrogen production of 1973, 1774, 1335, 1170 and 1070 mL H₂/L-OP sap, respectively, with the hydrogen content of 33.7, 29.6, 32.5, 31.5 and 26.4%, respectively. Identification of these high hydrogen producers using 16S rRNA sequence matching showed that the isolates PS-3 and PS-8 belonged to Clostridium beijerinckii, while the isolate PS-7 belonged to Clostridium acetobutylicum and the isolates PS-4 and PS-5 belonged to Klebsiella sp. and Klebsiella pneumoniae, respectively. Therefore, the pure culture C. beijerinckii PS-3 exhibited 1.8 folds higher hydrogen production (1973 mL H₂/L-OP sap) than the enriched cultures of 81RN1 (1085 mL H₂/L-OP sap).     

Biohydrogen production from dual digestion pretreatment of poultry slaughterhouse sludge by anaerobic self-fermentation

Poultry slaughterhouse sludge from chicken processing wastewater treatment plant was tested for their suitability as a substrate and inoculum source for fermentation hydrogen production. Dual digestion of poultry slaughterhouse sludge was employed to produce hydrogen by batch anaerobic self-fermentation without any extra-seeds. The sludge (5% TS) was dual digested by aerobic thermophilic digestion at 55 °C with the varying retention time before using as substrate in anaerobic self-fermentation. The best digestion time for enriching hydrogen-producing seeds was 48 h as it completely repressed methanogenic activity and gave the maximum hydrogen yield of 136.9 mL H₂/g TS with a hydrogen production rate of 2.56 mL H₂/L/h. The hydrogen production of treated sludge at 48 h (136.9 mL H₂/g TS) was 15 times higher than that of the raw sludge (8.83 mL H₂/g TS). With this fermentation process, tCOD value in the activated sludge could be reduced up to 30%.     

Comparison of UASB and EGSB reactors performance, for treatment of raw and deoiled palm oil mill effluent (POME)

Anaerobic digestion of palm oil mill effluent (POME) and deoiled POME was investigated both in batch assays and continuous reactor experiments using up-flow anaerobic sludge blanket (UASB) and expanded granular sludge bed (EGSB) reactors. The methane potential determined from batch assays of POME and deoiled POME was 503 and 610 mL-CH(4)/gVS-added, respectively. For the treatment of POME in continuously fed reactors, both in UASB and EGSB reactors more than 90% COD removal could be obtained, at HRT of 5 days, corresponding to OLR of 5.8 gVS/(L-reactor.d). Similar methane yields of 436-438 mL-CH(4)/gVS-added were obtained for UASB and EGSB respectively. However, for treatment of deoiled POME, both UASB and EGSB reactors could operate at lower OLR of 2.6 gVS/(L-reactor.d), with the methane yield of 600 and 555 mL-CH(4)/gVS-added for UASB and EGSB, respectively. The higher methane yield achieved from the deoiled POME was attributed to lower portion of biofibers which are more recalcitrant compared the rest of organic matter in POME. The UASB reactor was found to be more stable than EGSB reactor under the same OLR, as could be seen from lower VFA concentration, especially propionic acid, compared to the EGSB reactor.     

Extreme-thermophilic biohydrogen production by an anaerobic heat treated digested sewage sludge culture

Biohydrogen production process from glucose using extreme-thermophilic H₂-producing bacteria enriched from digested sewage sludge was investigated for five cycles of repeated batch experiment at 70 °C. Heat shock pretreatment was used for preparation of hydrogen-producing bacteria comparing to an untreated anaerobic digested sludge for their hydrogen production performance and responsible microbial community structures. The results showed that the heat shock pretreatment completely repressed methanogenic activity and gave the maximum hydrogen production yield of 355-488 ml H₂/g COD in the second cycle of repeated batch cultivation with more stable gas production during the cultivation when compared with control. Hydrogen production was accompanied by production of acetic acid. The average specific hydrogen in five cycles experiment ranged from 150 to 200 ml H₂/g VSS. PCR-DGGE profiling showed that the extreme-thermophilic culture predominant species were closely affiliated to Thermoanaerobacter pseudethanolicus.     

High yield simultaneous hydrogen and ethanol production under extreme-thermophilic (70 °C) mixed culture environment

The effect of pH and medium composition on extreme-thermophilic (70 °C) dark fermentative simultaneous hydrogen and ethanol production (process performance and microbial ecology) was investigated. Hydrogen and ethanol yields were optimized with respect to glucose, peptone, FeSO₄, NaHCO₃, yeast extract, trace mineral salts, vitamins, and phosphate buffer concentrations as well as initial pH as independent variables. A combination of low levels of both glucose (≤2 g/L) and vitamin solutions (≤1 mL/L) and high levels of initial pH (≥7), mineral salts solution (≥5 mL/L) and FeSO₄ (≥100 mg/L) stimulated the hydrogen production, while high level of glucose (≥5 g/L) and low levels of both initial pH (≤5.5) and mineral salts solution (≤1 mL/L) enhanced the ethanol production. High yield of simultaneous hydrogen and ethanol production (1.58 mol H₂/mol glucose combined with an ethanol yield of 0.90 mol ethanol/mol glucose) was achieved under extreme-thermophilic mixed culture environment. Results obtained showed that the shift of the metabolic pathways favouring either hydrogen or ethanol production was affected by the change in cultivation conditions (pH and medium composition). The mixed culture in this study demonstrated flexible ability for simultaneous hydrogen and ethanol production, depending on pH and nutrients formulation. The microorganisms involved could be regarded as simultaneous hydrogen/ethanol producers, as hydrogen and ethanol fermentation under all conditions was carried out by a group of extreme-thermophilic bacterial species related to Thermoanaerobacter, Thermoanaerobacterium and Caldanaerobacter.     

Biohydrogen production from cassava starch processing wastewater by thermophilic mixed cultures

Natural microbial consortia from hot spring samples were used to developed thermophilic mixed cultures for biohydrogen production from cassava starch processing wastewater (CSPW). Significant hydrogen production potentials were obtained from three thermophilic mixed cultures namely PK, SW and PR with maximum hydrogen production yields of 249.3, 180 and 124.9 mL H₂/g starch, respectively from raw cassava starch and 252.4, 224.4 and 165.4 mL H₂/g starch, respectively from gelatinized cassava starch. Acetic acid-ethanol and acetic-lactic acid type fermentation were observed in cassava starch fermentation, based on three thermophilic mixed cultures performance. The thermophilic mixed cultures PK, SW and PR exhibited the maximum hydrogen yield of 287, 264 and 232 mL H₂/g starch in CSPW, respectively corresponding to 53%, 48.7% and 42.8% of the theoretical values. Phylogenetic analysis of thermophilic mixed cultures revealed that members involved cassava starch degrading bacteria and hydrogen producers in both raw cassava starch and CSPW were phylogenetically related to the Thermoanaerobacterium saccharolyticum, Thermoanaerobacterium thermosaccharolyticum, Anoxybacillus sp., Geobacillus sp. and Clostridium sp.     

Early development of a shaftless horizontal axis wind turbine for generating electricity from air discharged from ventilation systems

The air discharged from ventilation systems is a high potential wind resource for generating electricity in countries where wind speed is unreliable or weak, such as in Thailand. The air discharged from ventilation systems produces consistent and high-speed wind when benchmarked against natural wind. However, the limitations of conventional wind turbines are that they have negative impacts on the ventilation system and are inconvenient to install in many areas. The innovative shaftless horizontal axis wind turbine (SHWT) introduced in this article has been designed to close the gap between the wind source and the conventional wind turbines in this process. The concept design shows how it could be mounted next to sources of waste wind, requiring only a small space for installation. An open hole is provided to enable airflow to be discharged into the environment. This SHWT has high market potential for utilizing man-made wind to generate electricity from an alternative source which supports sustainable energy development. The purpose of this study is to demonstrate the concept design of a prototype SHWT used for energy recovery from the discharged air of a ventilation system. How the rotor and stator design of the SHWT optimize wind turbine performance and minimize the negative effects on the ventilation system efficiency are also addressed in this study. The performance of the SHWT is demonstrated in a lab-scale test using the type of propeller fan that is generally applied in many sectors in Thailand. The results showed that the SHWT was successful in generating electricity and produced minimal negative effects on the ventilation system's performance. The maximum power output of the prototype SHWT is 7.4 W at a rotational speed of 1644 rpm using eight sets of magnets and 5.1 m/s wind speed. The maximum wind turbine efficiency is 51%. However, it still requires further optimization to enhance the SHWT performance.     

Biological activity of rice extract and the inhibition potential of rice extract,rice volatile compounds and their combination against α-glucosidase, α-amylase and tyrosinase

Rice is produced for consumption and traditional medicine. Rice is also used as an ingredient in cosmetic products. In this study, the author investigated the biological activity and inhibition potential against α-glucosidase, α-amylase and tyrosinase activity of rice extract (black rice [BR], red rice [RR] and white rice [WR]), rice volatile compounds, rice extract combined with volatile compounds, rice extract combined with standard inhibitors and volatile compounds combined with standard inhibitors. The results revealed that the free-radical scavenging capacity of rice extract is related to the phenolic content and flavonoids. BR showed the highest potential to inhibit α-glucosidase and α-amylase activity, whereas WR showed the highest potential to inhibit tyrosinase activity. Among rice volatile compounds, vanillin and vanillyl alcohol had the highest inhibition potential against α-glucosidase and α-amylase, respectively, whereas guaiacol had the highest inhibitory activity against tyrosinase. Molecular docking supported by the high binding efficiency was also obtained from vanillin and guaiacol when located at the active site of these enzymes. The combination of RR with acarbose (AB) had the highest inhibition potential and showed a synergic effect on both α-glucosidase and α-amylase. Interestingly, the combination of rice extract (BR, RR and WR) and vanillin and vanillyl alcohol had a synergic effect on α-amylase. Moreover, the combination of WR and vanillyl alcohol had the highest inhibition potential and showed a synergic effect on tyrosinase, whereas rice volatile compounds had a synergic effect on tyrosinase obtained from 2-pentylfuran/kojic acid (KA), vanillin/KA and vanillyl alcohol/KA.     

Effect of initial pH,nutrients and temperature on hydrogen production from palm oil mill effluent using thermotolerant consortia and corresponding microbial communities

Thermotolerant consortia were obtained by heat-shock treatment on seed sludge from palm oil mill. Effect of the initial pH (4.5–6.5) on fermentative hydrogen production palm oil mill effluent (POME) showed the optimum pH at 6.0, with the maximum hydrogen production potential of 702.52 mL/L-POME, production rate of 74.54 mL/L/h. Nutrients optimization was investigated by response surface methodology with central composite design (CCD). The optimum nutrients contained 0.25 g urea/L, 0.02 g Na₂HPO₄/L and 0.36 g FeSO₄·7H₂O/L, giving the predicted value of hydrogen production of 1075 mL/L-POME. Validation experiment revealed the actual hydrogen production of 968 mL/L-POME. Studies on the effect of temperature (25–55 °C) revealed that the maximum hydrogen production potential (985.3 mL/L-POME), hydrogen production rate (75.99 mL/L/h) and hydrogen yield (27.09 mL/g COD) were achieved at 55, 45 and 37 °C, respectively. Corresponding microbial community determined by the DGGE profile demonstrated that Clostridium spp. was the dominant species. Clostridium paraputrificum was the only dominant bacterium presented in all temperatures tested, indicating that the strain was thermotolerant.