Produzione di H2 e CH4 da frazioni biodegradabili di rifiuti urbani e da siero lattiero-caseario

Nowadays one of the main technical tasks is finding clean and renewable energy sources in order to overtake the dependence from fossil fuels. Bio-hydrogen and bio-methane production are considered interesting energetic alternatives, especially if the biological processes are applied to low cost and widely available substrates such as biodegradable wastes. Anaerobic digestion (AD) is a well-known technology to manage organic waste in an environmentally sustainable way, since it allows to achieve a certain level of stabilization of the organic matter and to exploit the energetic potential of the substrate; AD of biodegradable organic residues has therefore received renewed attention by the scientific and technical community over the last decades (Mata-Alvarez et al., 2000; Mata-Alvarez, 2002; De Baere, 2003; Bolzonella et al., 2006; Karagiannidis and Perkoulidis, 2009). If degradation of biodegradable organic substrates is appropriately operated in a two-staged mode (see e.g., Han and Shin, 2004a; Liu et al., 2006; Gómez et al., 2006, 2009; Ueno et al., 2007; Chu et al., 2008; Wang and Zhao, 2009; Lee et al., 2010b; Dong et al., 2011), separation of the acidogenic phase (producing through dark fermentation (DF) hydrogen and carbon dioxide as the gaseous products while releasing volatile fatty acids into the liquid solution) from the methanogenic stage (in which the residual and dissolved biodegradable organic matter from the first stage is finally converted into methane and carbon dioxide) can be accomplished. By means of the separation of the two biological phases, it is possible to obtain hydrogen besides methane gas, both to be exploited with various techniques. Considering the energetic aspect, combined H2 + CH4 production is, from a theoretical point of view, more favourable than conventional anaerobic digestion (Dong et al., 2011). Hydrogen gas, in particular, is considered a clean energy carrier characterized by the highest energy content per unit weight of any known fuel (142 kJ/g), and positive environmental implications from H2 utilization can be obtained from production through biological processes and from renewable sources. Various potentially biodegradable organic residues have been tested for fermentative H2 production, including food waste (FW), the organic fraction of municipal solid waste (OFMWS), and various agro-industrial residues, mainly due to their high carbohydrate content and wide availability (see e.g. Okamoto et al., 2000; Lay et al., 2003; Kim et al., 2004a; Liu et al., 2006; Alibardi et al., 2010, 2011; Kim et al., 2011a; Nazlina et al., 2011; De Gioannis et al., 2013). Whatever the substrate under concern, full-scale implementation of fermentative H2 production requires significant and stable generation rates. Since H2 is an intermediate by-product of anaerobic digestion and some fermentation reactions have low or null H2 production, identification of suitable conditions to harvest specific fermentative bacteria and/or inhibit the H2-consuming biomass is mandatory. Achieving optimal and stable H2 generation while keeping treatment costs low and producing an effluent suitable for further treatment, is probably the main technical issue to be addressed. To this regard, operational parameters including temperature, pH, presence of inoculum and its eventual pre-treatment, substrate to inoculum ratio (F/M), reactor configuration, substrate concentration, hydraulic retention time (HRT) and organic loading rate should be the subject for optimization of process efficiency. In particular, the influence of pH on fermentative H2 production is quite controversial in the literature. Generally, pH is a key parameter in fermentation that affects the degree of substrate hydrolysis, the activity of hydrogenase, as well as the metabolic pathways (Kim et al., 2011b). Changes in pH are thus reflected by variations in substrate and energy utilization, synthesis of proteins and various storage products, and metabolites production (Mu et al., 2006). In mixed anaerobic cultures, the operating pH also affects the relative proportions of microbial species (Fang and Liu, 2002; Shin and Youn, 2005; Mu et al., 2006; Nazlina et al., 2011). Highly acidic or basic pH values can negatively affect the activity of hydrogen-producing bacteria, since ATP is used to ensure cell neutrality rather than to produce H2 (Nazlina et al., 2011); low pHs can also result in inhibition of the hydrogenase activity (Khanal et al., 2004; Nazlina et al., 2011). Literature studies (see e.g., Yu et al., 2002; Khanal et al., 2004; Rechtenbach et al., 2008; Rechtenbach and Stegmann, 2009; Guo et al., 2010) generally agree upon the fact that H2-producing pathways involving acetate and butyrate production are favoured at pHs of approximately 4.5-6.0, while neutral or higher pHs promote ethanol and propionate production (associated to either H2-neutral or H2-consuming pathways). The effects of pH on the metabolic pathways and the associated fermentation yield have also been reported to depend on the specific characteristics of the inoculum and substrate (Nazlina et al., 2011), in particular for complex substrate compositions. The results of studies on the effect of pH on H2 production often show inconsistencies due to the different experimental conditions (substrate/inoculum type, amount and pre-treatment method, pH control strategy), so that systematic investigations on this specific issue are still missing. Also, the effects of the ratio between substrate and inoculum also indicated as F/M (food to microorganism ratio), S/I or S/X (substrate to inoculum ratio), I/S and ISR (inoculum to substrate ratio) are poorly documented for anaerobic digestion of FW (Boulanger et al., 2012) and only few works are conducted specifically on dark fermentation of FW. Pan et al. (2008) has reported the effect of F/M (food to microorganism ratio) on hydrogen production of mixed food waste, finding that the optimal F/M ratio in mesophilic conditions was 6, with an H2 production of 39 ml/g VS. Further increase of the F/M resulted in diminished H2 production. During the present research activity, combined H2 and CH4 production from two residues having different characteristics, cheese whey (CW) and municipal solid waste organic fraction (OFMSW), was studied. Various factors (in particular pH and F/M ratios) were studied in order to enhance H2 and CH4 yields, in particular to assess the optimal operating conditions for fermentative hydrogen production from the two substrates considered. As far as OFMSW was concerned, also the possibility of producing a compost of good quality from the solid phase of the anaerobic digestate was assessed. In the first part of the research, the activity focused on CW treatment. Cheese whey is a by-product of cheese manufacturing and its main components are lactose (45-50 g/l), proteins (6-8 g/l), lipids (4-5 g/l) and mineral salts (8-10% dried extract). The CW treated in this study, is the effluent from cheese production using a mixture of sheep and cow milk. Preliminary dark fermentation batch tests confirmed that hydrogen production from CW is possible even without any specific inoculum and strongly affected by multiple factors including substrate characteristics, as well as pH. Afterwards, semi-continuous anaerobic digestion tests were performed obtaining a stable hydrogen and methane production. For the first stage, three test were conducted, under mesophilic conditions with an HRT of 2 days, at the following pHs (7.5; 6.5; 5.5). The test operated with pH continuously controlled at 5.5 reached the maximum average yield of 80.8 Nl H2/kg TOC (38.4 Nl H2/kg VS) while in the second stage, operating under mesophilic conditions without pH control and adopting a HRT of 10 days, the average yield was 660.7 Nl CH4/kg TOC (383.6 Nl CH4/kg VS). The second part of the research activity was devoted to the OFMSW treatment. OFMSW may represent a relatively inexpensive and suitable source of biodegradable organic matter mainly due to its high carbohydrate content and wide availability (Okamoto et al., 2000; Lay et al., 2003; Kim et al., 2004; Liu et al., 2006; Li et al., 2008a; Li et al., 2008b; Zhu et al., 2008; Wang and Zhao, 2009; Kim et al., 2011a; Nazlina et al., 2011). Synthetic OFMSW was produced in laboratory, in order to make easier the comparison between the results of different tests, according to the following recipe: 8% of meat, 72% of fruit and vegetables, 8% of bread and 12% of cooked pasta (on a wet weight basis). Batch dark fermentation tests using untreated activated sludge as source of inoculum (T= 39°C; pH = 6.5) were performed at four F/M ratios (4; 7; 12 and 18) in order to assess the optimal one. The optimal F/M ratio resulted to be 7, corresponding to the maximum H2 yield of 89.9 Nl H2/ kg VS. Then, the effluent of the first stage (operated with the optimal F/M) was fed, mixed with methanogenic inoculum (F/M ratio = 0.5), to the second stage reactor to investigate methane production. The attained H2 and CH4 production was compared to the CH4 production achieved with a conventional single-stage batch reactor. The results confirm an higher CH4 yield for the second stage, 354 Nl CH4/kg VS versus 271 Nl CH4/kg VS for the single-stage. After a detailed analysis of the informations obtained from the batch tests, a double stage system was operated under semi-continuous conditions in order to better approximate the full scale operation. In the first semi-continuous stage, (T = 39°C; HRT = 2 days; pH = 6.5; F/M = 18), the average yield was 31,8 Nl H2/kg VS while in the second one a stable methane production of 432 Nl CH4/kg VS was obtained at HRT = 25 days (T = 39°C; pH not set). Finally, the solid phase of the digestate coming out form the double stage operated under semi-continuous conditions was composted after dewatering and mixing with a bulking agent. The final product was analysed for the main compost properties. The obtained compost reached a good stability (BOD4= 2 mg O2/l) and did not showed problems of phytotoxicity (germination index = 50%).