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Sumathy, “An overview of hydrogen production from biomass”, Fuel Processing Technology 87 (2006) 461 – 472
The phenomenon of biological hydrogen production was observed one century ago. When the oil crisis broke out in 1970s, the technology started receiving attention, especially in hydrogen production by photosynthetic process. However, these works are in laboratory scale and the practical applications still need to be demonstrated. Biological hydrogen production can be classified into five different groups: (i) direct biophotolysis, (ii) indirect biophotolysis, (iii) biological water–gas shift reaction, (iv) photo-fermentation and (v) darkfermentation . All processes are controlled by the hydrogen-producing enzymes, such as hydrogenase and nitrogenase. The major components of nitrogenase are MoFe protein and Fe protein. Nitrogenase has the ability to use magnesium adenosine triphosphate (MgATP) and electrons to reduce a variety of substrates (including protons). This chemical reaction yields hydrogen production by a nitrogenase-based system : where ADP and Pi refer to adenosine diphosphate and inorganic phosphate, respectively. Hydrogenases exist in most of the photosynthetic microorganisms and they can be classified into two categories: (i) uptake hydrogenases and (ii) reversible hydrogenases. Uptake hydrogenases, such as NiFe hydrogenases and NiFeSe hydro-genases, act as important catalysts for hydrogen consumption as follows: Reversible hydrogenases, as indicated by its name, have the ability to produce H2 as well as consume H2 depending on the reaction condition, Koku et al.  have reported that though a variety of substrates can be used for Rhodobacter sphaeroides growth, only some of them are suitable for hydrogen production. The properties of nitrogenase and hydrogenase are summarized in Table 4. 4.2. Direct biophotolysis Direct biophotolysis of hydrogen production is a biological process using microalgae photosynthetic systems to convert solar energy into chemical energy in the form of hydrogen:
Hallenbeck and Benemann  performed similar cost estimation and reported that the capital cost of US$100/m2. such as green algae and Cyanobacteria (blue-green algae). The works reported on mutants for hydrogen production are listed in Table 5 [70– 74]. using mutants for hydrogen production. The electrons are then transferred to the ferredoxin (Fd) using the solar energy absorbed by PSI. such as gas separation and handling. Since hydrogenase is sensitive to oxygen. microalgaes. This condition can be obtained by the use of green algae Chlamydomonas reinhardtii that can deplete oxygen during oxidative respiration . In their estimation. 5: (i) biomass production by photosynthesis. and (iv) conversion of 2 mol of acetates into hydrogen. mutants derived from microalgae were reported to have good O2 tolerance and thus higher hydrogen production. some practical factors were neglected. due to the lack of hydrogenase. have the ability to produce hydrogen. Cyanobacteria are used to produce hydrogen via the following reactions : .3. 4. Benemann  estimated the cost of direct biophotolysis for hydrogen production to be $20/GJ assuming that the capital cost is about US$60/m2 with an overall solar conversion efficiency of 10%. (ii) biomass concentration. only CO2 reduction takes place. In green plants. it is necessary to maintain the oxygen content at a low level under 0. contain hydrogenase and. Indirect biophotolysis According to Gaudernack . electrons are generated when PSII absorbs light energy. The hydrogenase accepts the electrons from Fd to produce hydrogen as shown in Fig. (iii) aerobic dark fermentation yielding 4 mol hydrogen/mol glucose in the algae cell. two photons from water can yield either CO2 reduction by PSI or hydrogen formation with the presence of hydrogenase. thus. In the biophotolysis process. Recently.1% so that hydrogen production can be sustained . along with 2 mol of acetates. the concept of indirect biophotolysis involves the following four steps as illustrated in Fig. It can be seen that.Two photosynthetic systems are responsible for photosynthesis process: (i) photosystem I (PSI) producing reductant for CO2 reduction and (ii) photosystem II (PSII) splitting water and evolving oxygen. the efficiency is low. On the contrary. In this process. In a typical indirect biophotolysis. the efficiency can be increased significantly. 4. However. due to the significant amount of substrate being respired and consumed during this process.
However.4. including CO dehydrogenase. The common objectives of these works were to identify suitable microorganisms that had high CO uptake and to estimate the hydrogen production rate. such as R. The estimated overall cost is US$10/GJ of hydrogen . An alternative new chemoheterotrophic bacterium Citrobacter sp. However.  investigated the indirect biophotolysis with Cyanobacterium anabaena variabilis exposed to light intensities of 45–55 Amol_1 m_2 and 170–180 Amol_1 m_2 in the first stage and second stage. The hydrogen production rate through indirect biophotolysis is comparable to hydrogenase-based hydrogen production by green algae. Biological water–gas shift reaction Some photoheterotrophic bacteria. . rubrum was less than 5 h by the oxidation of CO to CO2 coupled with the reduction of protons to hydrogen. and the gram-positive bacteria. Organisms growing at the expense of this process are the gramnegative bacteria. .3 yielded optimal hydrogen production.8 and 8. anaerobic conditions in the presence of sufficient nickel. the doubling time of R. rubrum and Rubrivax gelatinosus.5 ml H2/gcdw h (cdw: cell dry weight) was found. the dominating products are CO2 and H2. Biological water–gas shift reaction for hydrogen production is still under laboratory scale and only few works have been reported. Photoproduction of hydrogen at a rate of about 12. Under anaerobic conditions. R. this process is favorable for hydrogen production. Therefore.  observed that under dark.Markov et al. Fe–S protein and COtolerant hydrogenase. Kerby et al.2 atm. The estimated cost is subject to a significant change depending on the technological advancement. Y19 was tested by Jung et al. such as Rhodospirillum rubrum can survive in the dark by using CO as the sole carbon source to generate ATP by coupling the oxidation of CO to the reduction of H+ to H2 : In equilibrium. it was found that maintaining the medium at pH value between 6. 4. In the study on indirect biophotolysis with Cyanobacterium gloeocapsa alpicola by Troshina et al. respectively. CO induces the synthesis of several proteins. rubrum requires light to grow and hydrogen production is inhibited by medium CO partial pressure above 0. Increasing the temperature from 30 -C to 40 -C can increase the hydrogen production twice as much. it should be pointed out that indirect biophotolysis technology is still under active research and development. Electrons produced from CO oxidation are conveyed via the Fe–S protein to the hydrogenase for hydrogen production . such as Carboxydothermus hydrogenoformans .
5.8/GJ) for a methane concentration between 1% and 10%. for hydrogen production using water–gas shift reaction.6/GJ) to around US$2. the products of dark fermentation are mostly H2 and CO2 combined with other gases. Unlike a biophotolysis process that produces only H2. hydrogen can be produced by photo-fermentation of various types of biomass wastes. Their analysis showed that biological water–gas shift process was economically competitive when the methane concentration was under 3%. The hydrogen production cost from biological water– gas shift reaction ranged from US$1. 4. 4. such as green algae on carbohydrate-rich substrates. This process using dark fermentation for hydrogen production is illustrated in Fig.25/kg (US$18. the cost of biological water–gas shift processes are lower due to the elimination of reformer and associated equipment. Wolfrum et al. However. as shown in Fig. depending on the reaction process and the substrate used. some attempts have been made for hydrogen production from industrial and agricultural wastes to effect waste management. rubrum. Photo-fermentation Photosynthetic bacteria have the capacity to produce hydrogen through the action of their nitrogenase using solar energy and organic acids or biomass. photo-fermentation process is not a competitive method for hydrogen production.6. Dark fermentation Fermentation by anaerobic bacteria as well as some microalgaes. In recent years. 7.  have conducted a detailed study to compare the biological water–gas shift reaction with conventional water–gas shift processes. such as CH4 or H2S. Recently. With glucose as the model substrate. The maximum hydrogen production activity was found to be 27 mmol/g cell h. which is about three times higher than R. at the present time. maximum 4 mol H2 is produced per mole glucose when the end product is acetic acid: . 6. these processes have three main drawbacks: (i) use of nitrogenase enzyme with high-energy demand. As summarized in Table 6 [84–87]. Hence. This process is known as photofermentation. can produce hydrogen at 30 -C to 80 -C especially in a dark condition . (ii) low solar energy conversion efficiency and (iii) demand for elaborate anaerobic photobioreactors covering large areas .75/kg (US$14. Compared with thermochemical water– gas shift processes.
When hydrogen concentration increases. Chang b. hydrogen production rate was reduced from 198 to 34 mmol l_1 day_1. hydraulic retention time (HRT) and gas partial pressure.W Chang a. C. which in turn decrease the hydrogen production . Freezing and thawing and sterilization increased the specific hydrogen yield by 1.P. . C. C. sterilization. such as lactate. A much higher hydrogen yield than presented in the literature was obtained.C. Also. Hence. Partial pressure of H2 is yet another important parameter affecting the hydrogen production.5_/2.5 day to 3 days. This work examined the anaerobic digestion of wastewater sludge using a clostridium strain isolated from the sludge as inoculum. HRT also plays an important role in hydrogen production. Wang a. B. the 4 mol H2 production/mol glucose cannot be achieved because the end products normally contain both acetate and butyrate . For the optimal hydrogen production. butanol or alanine. the feasibility of the technology yields a growing commercial value. The amount of hydrogen production by dark fermentation highly depends on the pH value. Due to the fact that solar radiation is not a requirement. acidification. in practice.*. Ueno et al.5 day could effect maximum hydrogen production (14 mmol/g carbohydrate) from wastewater by anaerobic microflora in the presence of chemostat culture. D. Liao b „ Producing hydrogen from wastewater sludge by Clostridium Bifermentans „ . while adding an inhibitor and ultrasonication reduced the hydrogen yield.However.5 times to that of untreated sludge. the metabolic pathways shift to produce more reduced substrates. Besides the pH value and partial pressure. Chu a. freezing/thawing and adding methanogenic inhibitor*/on the production of hydrogen were examined. When HRT was increased from 0. while the carbohydrates in the wastewater were decomposed at an increasing efficiency from 70% to 97%. acetone. C.S. However.-V. the effects of five pre-treatments*/ultrasonication.J.  have reported that an optimal HRT of 0. Journal of Biotechnology 102 (2003) 83_/92 Excess wastewater sludge collected from the recycling stream of an activated sludge process is biomass that contains large quantities of polysaccharides and proteins. ethanol. hydrogen production by dark fermentation does not demand much land and is not affected by the weather condition. Lee a. relevant literature indicates that the bio-conversion of wastewater sludge to hydrogen is limited and therefore not economically feasible. pH should be maintained between 5 and 6 [90– 92].
On the other hand. Sweet sorghum biomass is rich in readily fermentable sugars and thus it can be considered as an excellent raw material for fermentative hydrogen production. Sweet sorghum is an annual C4 plant of tropical origin. Vol 9. containing the cellulose and hemicelluloses. Hydrogen productivity of sorghum residues fermented with R.86 mol hydrogen per mol of glucose consumed. ANGELOPOULOS. SKIADAS. an important. This corresponded to a hydrogen productivity of 10.. G. . albus.N.6 mol hydrogen per mol of glucose consumed.I. fibrolytic bacterium of the rumen. albus was as high as 2. at a hydraulic retention time of 12 hours. The highest hydrogen yield obtained from sorghum extract fermented with mixed microbial cultures in continuous system was 0. ANTONOPOULOU.1-2. pp 144-151. coming from the indigenous sorghum microflora and Ruminococcus albus. Extraction of free sugars from the sorghum stalks was achieved using water at 30°C. a liquid fraction (sorghum extract).6 mol hydrogen per mol of glucose consumed.4 l hydrogen per kg of sorghum biomass and was comparable with those obtained from batch experiments. „BIOHYDROGEN PRODUCTION FROM SWEET SORGHUM BIOMASS USING MIXED ACIDOGENIC CULTURES AND PURE CULTURES OF RUMINOCOCCUS ALBUS” . and a solid fraction (sorghum cellulosichemicellulosic residues). were obtained. I. NTAIKOU. the hydrogen yield obtained from sorghum extract treated with R. In total. albus reached 2. Hydrogen production from sorghum extract was investigated using mixed acidogenic microbial cultures. LYBERATOS. using R. the productivity of sorghum biomass (that of sorghum extract plus that of sorghum residues) could be 60 l hydrogen per kg of sorghum biomass if R. well-adapted to sub-tropical and temperate regions and highly productive in biomass. Hydrogen productivity of sorghum residues was assessed as well. 2007 The present study focuses on the exploitation of sweet sorghum biomass as a source for hydrogen in continuous and batch systems.V. No 2. After the extraction process.K. rich in sucrose. Global NEST Journal. albus is used.G. H. GAVALA.
. 2010).. Biotechnol.. 2009.Laurent Beckers. 1995..molhexose) but can achieve higher production rates (Hallenbeck et al. 2004. 2009. facultative anaerobes such as Enterobacteriaceae that present lower experimental yields (~ 1 molH2. Moriarty et al. One of the most promising and investigated prospects is the biological production of hydrogen through the degradation of a large spectrum of carbon sources by anaerobic bacteria in a process called “dark fermentation” (Das et al.. strict anaerobic bacteria from the genus Clostridium that have the potential to reach high experimental hydrogen yields (about two moles of hydrogen per mole of hexose consumed). there has been an increasing interest to find new H2 production processes with almost no carbon emission (Balat. sucrose. Hydrogen may evolve through the fermentation processes of simple carbohydrates such as glucose. 2008). 2008a). Kraemer et al.The best described mesophilic strains are. lactose and maltose or more complex ones such as starch or even cellulose (Ueno et al. Agron.. Clostridia are however extremely sensitive to the presence of oxygen which strongly inhibits H2 evolving enzymes (Heinekey. 2002. 541-548 Hydrogen (H2). The two species investigated in this work. Yuan et al. Das.. Davila. This can be avoided with the addition of an expensive reducing agent such as L-cysteine. 2010 14(S2). Clostridium butyricum CWBI1009 (Masset et al. Nevertheless.. 2004a. „Fermentative hydrogen production by Clostridium butyricum CWBI1009 and Citrobacter freundii CWBI952 in pure and mixed cultures”. a process releasing large amount of fossil CO2 in the atmosphere.. Hallenbeck. Oh et al.. 2009. Soc. Hawkes et al. 2008)... 2010) and Citrobacter freundii CWBI952 (Hamilton et al. is a very promising clean energy vector for the decrease of our environmental impact since its utilization generates only water vapor.. Only a few studies have investigated the hydrogen production with these different substrates on pure cultures in . 2009). 2009). whether burned or used directly in a fuel cell. H2 is still mainly produced by steam reforming of methane. 2009). The main purpose to enhance fermentative hydrogen production is to improve hydrogen yields for an efficient energy recovery from the substrate. Nath et al. Christopher Hamilton. 2002. And on the other hand.. 2008. on the one hand.. 2008). Holladay et al. However. the use of such an agent is not suitable for a large-scale cost effective biohydrogen production process (Das et al. Magnusson et al. Serge Hiligsmann. Philippe Thonart.Vazquez et al. Levin et al. Environ. Nath et al. In the last few years.molhexose -1 respectively depending on the metabolic pathway followed for the fermentation of the carbon source (Nandi et al. Kotay et al. 2008... 2007.. Julien Masset. have a maximum theoretical hydrogen yield of 4 and 2 molH2.. 2004b. 2001. 1998.
1998. However.. A. 2006. butyrate. 2002). 1995. For testing fermentative bio- . A. The biological methods for generating H2 include light-dependent methods. Biological methods of hydrogen production are preferable over chemical methods because they offer the possibility of using sunlight. butyricum growth. Nath et al. acetate. Fernández. butyricum growing on starch (Yokoi et al. formate. 2009). Pan et al. ethanol and succinate) in pure C. little is known about this consortium on other substrates. This has already been shown in a mixed culture of Enterobacter sp. 2007). This is why this work compares the hydrogen and major metabolites production (i. Bioresource Technology 102 (2011) 8621–8627 Conventional methods for producing H2 gas include: steam reforming of methane and hydrocarbons.e. M. which were observed in the past because of an expectation of increasing demand for raw materials. freundii and Cl. Morán. „Hydrogen production: Two stage processes for waste degradation”. Such a culture would not require the addition of any reducing agents since C. as also biomass pyrolysis and gasification through the steam reforming of gases by water–gas shift reactions. These methods are considered environmentally benign conversions.. Fierro. it is worthwhile mentioning the production of H2 by thermal processing of biomass. 2008.. These experiments were carried out in serum bottles batch cultures based on the biochemical hydrogen potential (BHP) test procedure described by Lin (Lin et al. when it is taken into account that a continuous increase is expected in the demand for the use of H2. J. Sánchez. (2007) compared two different biological methods for hydrogen production: fermentative and photosynthetic based upon the modality of batch cultures. freundii and Cl. 2008).. C. However. In this work. the need of light of this latter process for microbial growth and the use of CO as a carbon source may seem to be technical and economic barriers with regard to the reactor design and the thermal pre-treatment step necessary for the generation of CO.. and Cl. In this way. butyricum in the same BHP culture may enhance hydrogen production. bioelectrochemical systems (BES) and water–gas shift reaction mediated by photoheterotrophic bacteria. Freundii consumes oxygen and provides the anaerobic conditions required for Cl. thus avoiding market distortions.comparison with co-cultures (Yokoi et al. In this category are included the dark fermentation process. the main goal in the near future should be to attain cost-effective production processes from renewable sources. They are discussed in comparison with the results found in the literature. non-catalytic partial oxidation of fossil fuels and auto-thermal reforming.E. butyricum cultures with five different substrates. a co-culture of C. Gómez !. lactate. such as direct and indirect biophotolysis and photo-fermentation.. which take place under moderate conditions (Redwood et al. These methods present the main advantage of being capable of treating biomass with a high lignocellulosic content. CO2 and organic wastes as substrates. X. Escapa. Furthermore.. Among the processes currently available to fulfil this objective. cocultures were monitored on glucose and also on starch for comparison. The other routes for biological production are not light-dependent methods. Ustak et al. Chen et al. These methods are all energy intensive processes requiring high temperatures (>850 "C) (Kapdan and Kargi. 2006).
Several disadvantages of these methods have been stated by Das and Vezirog˘lub (2001). The process is then characterised by low efficiency and this also becomes the main limiting factor for commercial application. some of these pre-treatment options may increase operating costs. Improving H2 yields Strategies for improving H2 yields have been reviewed by Kraemer and Bagley (2007) analysing diverse alternatives. The testing of green algae proved that the most effective was the algae species Scenedesmus. considering the composition of the effluent stream of the dark fermentation process. this acting as a barrier for commercial application. As dark fermentation has a maximum yield of 33% (on sugars). Thus. the appearance of caproate in the final products is an indication that significant solventogenesis has occurred and thus the yield of the fermentative H2 production is poor (Ding et al. when considering large scale implementations. Several options successfully tested for obtaining active hydrogen producing microflora are listed in Table 2. butyrate) (Bartacek et al. direct biological production of H2 appears to be restricted to a pretreatment step in a larger bio-energy or biochemical production concept (Angenentet al. In the present review. The fermentative method of H2 production was more efficient. a description is also presented of possible second stage processes for further degradation of dark fermentation effluents (mainly containing fatty acids). either ethanol and acetate or ethanol and butyrate. has a longer efficiency of a single charge and enables effective use of different biological wastes. 2007). 2010). A fact of great relevance is the small scale at which most of these technologies have been tested. Alternatives for the second stage considered were photofermentation and BES as processes capable of converting fermentation sub-products into H2.. Thus. which makes the process an alternative for the conventional biological methods for treating and taking full advantage of this biomass. These pre-treatments may regularly be applied to reactors in order to maintain the activity of H2 producing microorganisms. High bio-hydrogen purity was analytically verified. with the dark fermentation process being the only one with a prospect of being rapidly scaled up owing to its similarity to the well-known anaerobic digestion process. In addition. However. production of hydrogen from either of the methods previously commented upon is characterised by low efficiency. Nonetheless. this process can use residual carbohydraterich biomass as substrate. Anaerobic digestion as a final stabilisation stage was also considered. differences in hydrogen yields have proved to disappear in continuous experimentations which indicate that pre-treatment methods have only short-term effects on hydrogen production (Luo et al. The remainder is mainly composed of volatile fatty acids (VFAs – acetate. such as inoculum pre-treatment. 2004). However.. four mixed cultures representing anaerobic microorganisms (dominant strain Clostridium) were selected. a description of successful experiences for the production of hydrogen through dark fermentation from residual biomass was undertaken. Another relevant aspect that should be considered is . with the presence of this acid being coupled to the production of H2.. In addition. 2010).hydrogen production. The process itself may be seen as a pretreatment step in a complete stabilisation chain. the formation of caproate has also been reported in dark fermentation systems. (2006) for maintaining an active H2 producing population during extended operation. gas sparging. Only 33% of the chemical oxygen demand (COD) contained in the wastecan be transformed into hydrogen (considering glucose). it does not need light. as it is the case of heat shock on startingup the reactor or from ongoing heat treatment as proposed by Khanal et al.. reduction of H2 and CO2 levels in the liquid phase. The generation of caproate does not translates into a higher H2 yield since its formation has been explained by the secondary fermentation of two substrates. owing to the wide application of this technology in the treatment of bio-wastes. and varying organic loading rates.
fluctuating H2 production has also been reported.. accumulation of VFAs still poses a problem. (2005). either gas sparging or vigorous mixing will aid in reducing H2 and CO2 concentrations inside the reactor (Kraemer and Bagley. owing to VFA accumulation. 2009.. (2009) reported stable operation during continuous fermentation of household waste when mixing was provided to the reactor in contraposition to what occurs in static reactors. Microbial consortia may address these issues if they are selected for growth and dominance under non-sterile conditions (Hallenbeck and Ghosh. Jung et al. Gómez et al. 2008). Laboratory and pilot scale fermentation experiences Centralised collection of urban solid wastes and chemical characteristics of the organic fraction contained in these wastes are two factors that make them a suitable substrate for the dark fermentation process. 2009). This same effect may be attained by gas sparging inside the reactor. Lee et al. These divergences in results obtained may indicate that the selection of optimum parameters is a key factor for attaining stable performance.. Similar results were reported by Gómez et al. Immobilized systems are an effective and stable approach for continuous hydrogen production allowing efficient utilization of carbon substrates (Jo et al.. 2005. in an attempt to increase the treatment capacity of reactors may lead to inhibition of the fermentation system. 2008. attached growth systems are used for the fermentation of high strength wastewater. with this being detrimental to H2 yield. Several reports in litter-scale can be found in literature. This is the case of results obtained by Zhu et al. Another feedstock of interest is the use of marine algae. with Laminaria japonica presenting the highest H2 yields. However. adaptation of the inoculum may play a crucial role in attaining stable operation at high concentrations of VFAs. when food wastes were treated as the substrate.. Nguyen and co-workers (Nguyen et al. 2007) and cheese whey (Ferchichi et al. Operating alternatives associated with the removal of such products will favour process efficiency.. Wang and Zhao (2009) demonstrated that a significant reduction in H2 yield was caused by an increase of the OLR to 30. Substrates such as wastewaters with high carbohydrate content have also been demonstrated to be suitable for the dark fermentationfermentation process. 2008). Experiments with dark fermentation processes are rapidly increasing on a laboratory scale. On these lines. (2009) when fermenting food waste under static conditions. 2010) under different temperature conditions. (2011) tested various algae for fermentative hydrogen production. Kim et al. Liu et al. Chu et al.2 kg VS m!3 d!1. increases in H2 yields being reported when there were increases in temperature to thermophilic regimes. while in general. which have been studied using completely stirred tank reactors (CSTRs) and reactors with immobilised biomass. 2009.. as demonstrated by Valdez-Vazquez et al. fermentation of household wastes has been studied by several authors (Gómez et al. However.the inhibition caused by the accumulation of fermentation products. 2010) demonstrated that the removal of the H2 produced from the gas headspace during batch fermentation improved H2 yields. Steady operation over long periods of reactors producing H2 has been reported by several authors (Valdez-Vazquez et al.. In this sense. In addition. Table 3 shows a list of published .. the fermentation of wastes for H2 production may be considered as a pre-treatment option which may be integrated with minimum modifications into centralised solid waste treatment plants where an anaerobic digester is already operating. An increase in the organic loading rate. These authors reported successful performance from a reactor treating organic wastes during solid state fermentation. The presence of particulate material set limits to the application of high-rate systems. This is the case for molasses (Li et al. 2006) and thus favour stability for extended operating periods. The commercialization of industrial hydrogen fermentation makes imperative to achieve steady operation and also to carry out the process under non-sterile conditions using readily available complex feed stocks with only minimal pre-treatment. (2008) when fermenting potato waste in a two-phase configuration. For this reason CSTR systems are often applied for the treatment of household wastes. In this way.. 2005).
This was the case for the two-phase process (thermophilic-mesophilic) with sludge recirculation tested by Lee et al. the effects on a potential second treatment process should be considered. since lower H2 productivity was attained during recycling. such as the lack of suitable sugars in the wastewater. (2009) studied the fermentation of swine manure supplemented with glucose in a 4 L working volume reactor under mesophilic conditions at varying pH and HRT. This configuration presented two main advantages. Addition of precipitated sludge at the bottom of a storage tank was used for pH control. but also leads to an effluent with high conductivity. On these same lines. These authors obtained stable operation over a period of 150 days with a H2 yield of 205 mL g!1 VS added. 2009). Although results are promising.results for different fermentation systems working under continuous operation in reactors of size bigger than 20 L. Although results reported were not satisfactory. single fermentation of swine manure resulted in low to negative H2 yields. this strategy has been tested with satisfactory results by other authors. There are several factors that may contribute to low hydrogen production from swine manure. Another option for reducing the amount of alkaline solutions needed would be the use of co-substrates. since most of the hydrogen evolved in high-rate hydrogen fermentation tests is a result of sugar in the sample. which not only increases operating costs. These authors reported that to increase hydrogen content in the offgas.. Kraemer and Bagley (2005) proposed the recycling of a fraction of the methanogenic effluent in order to take advantage of the alkalinity generated in the digestion phase which is needed in the dark fermentation phase for pH control. Perera and Nirmalakhandan (2010) obtained successful results under batch conditions when co-fermenting sucrose with heattreated cattle manure. (2010) in a 10 L H2 reactor and a 40 L CH4 reactor. although high VFA concentrations might occur. These authors reported a 10% improvement in the H2 yield and also demonstrated the capacity of these wastes to reduce buffering needs. Controlling the pH of the process requires large amounts of alkaline solutions. on feature that needs to be addressed is the requirement for alkalis. so that the reagent for pH adjustment could be saved. The second advantage was that acidity in the hydrogen production stage was neutralized by recirculation of the digested sludge. On the basis of the results recorded above. It appears that hydrogen recovery from swine wastewater will not be feasible by fermentation processes unless some breakthrough is made in changing the nature of the wastewater or the conditions for microbial growth that inhibit the utilization of H2 by microorganisms in the wastewater (Wagner et al. . either to enhance treatment efficiency or to make the overall process economically viable by the generation of a high-value product in a second fermentation stage (Mohan et al. 2010). The co-fermentation of manures and carbohydrate-rich wastes would avoid acidification of the reactor. which presented high alkalinity. When concentrated solutions of NaOH are used.. Zhu et al. The first was that the hydrogen-producing bacteria which exist in digested sludge could replenish the hydrogen production reactor by recirculation. it seems reasonable to assume that different strategies intended to increase fermentation yields and lower operating costs should also consider the use of acid-rich effluents. However. methane production had to be limited below 2%.