DGGE analysis of buffalo manure eubacteria for hydrogen production: effect of pH, temperature and pretreatments.

Buffalo dung is a low-cost substrate with plenty of carbohydrates, an optimal carbon/nitrogen ratio, and a rich microbial flora, and could become a valuable source of biogas. Therefore, in the present study we compared the type and amount of specific eubacteria to the different configurations of pH, temperature and thermal pretreatment after fermentation in batch reactors in order to understand the suitability of buffalo manure for hydrogen production. The phylogenetic structure of the microbial community in fermentation samples was studied using denaturing gradient gel electrophoresis to generate fingerprints of 16S rRNA genes. The sequences analysis revealed abundance of the phyla Bacteroidetes and Firmicutes, and in particular of the order Clostridiales. Very active hydrogen producing bacteria belonging to Clostridium cellulosi species were identified demonstrating the suitability of this substrate to produce hydrogen. Moreover, a large fraction of 16S-rDNA amplicons could not be assigned to lower taxonomic ranks, demonstrating that numerous microorganisms involved in anaerobic fermentation in digesters or bioreactors are still unclassified or unknown.

The peculiar role of C/N and initial pH in anaerobic digestion of lactating and non-lactating water buffalo manure.

Manure from lactating and non-lactating water buffaloes was separately collected from a single dairy farm and anaerobically digested under mesophilic conditions in batch mode to produce biogas. This substrate, scarcely studied in the literature, showed two peculiarities regarding two fundamental parameters in the digestion processes: C/N ratio and initial pH. Typically, optimal C/N varies from 20 to 30, but in this work an almost negligible role of this ratio is observed. We demonstrated it by investigating a very large C/N interval, from 9.7 to 50.1, not by adding selected nutrients to the system, but exploiting the natural variation of the substrate. Concerning the pH, we show that also typically considered unfavorable conditions are feasible for this substrate. In fact, though neutral-basic initial pH is proved to be optimal to run the digestion process, in line with many other kinds of dungs, also acid initial pH leads to satisfactory CH4 yield. This is principally related to the capability of water buffalo manure of auto-modifying the pH to neutrality during the digestion, when initial pH of 5.0 and 6.0 are considered. This aspect may be relevant in co-digestion processes with acid wastes, since it may allow not adding neither a buffer, nor a pH regulator to the system. All the digestion conditions are separately tested with lactating and non-lactating water buffaloes and no statistical meaningful differences exist between the two kinds of cattle.