Prokaryotic population dynamics and interactions in an AnSBBR using tequila vinasses as substrate in co-digestion with acid hydrolysates of Agave tequilana var. azul bagasse for hydrogen production

Abstract

Aims: The purpose of this study was to characterize the prokaryotic community and putative microbial interactions between H₂ -producing bacteria (HPB) and non-HPB using two anaerobic sequencing batch biofilm reactors (AnSBBRs) fed with tequila vinasses in co-digestion with acid hydrolysates of Agave tequilana var. azul bagasse (ATAB).

Methods and results: Two AnSBBRs were operated for H₂ production to correlate changes in physicochemical and biological variables by principal component analysis (PCA). Results indicated that H₂ yield was supported by Ethanoligenens harbinense and Clostridium tyrobutyricum through the PFOR pathway. However, only E. harbinense was able to compete for sugars against non-HPB. Competitive exclusion associated with competition for sugars, depletion of essential trace elements, bacteriocin production and resistance to inhibitory compounds could be carried out by non-HPB, increasing their relative abundances during the dark fermentation (DF) process.

Conclusions: The global scenario obtained by PCA correlated the decrease in H₂ production with the lactate:acetate molar ratio in the influent. At the beginning of co-digestion, this ratio had the minimum value considered for a net gain of ATP. This fact could cause the reduction of the relative abundance of C. tyrobutyricum.

Significance and impact of the study: This is the first study that demonstrated the feasibility of H₂ production by Clostridiales from acid hydrolysates of ATAB in co-digestion with tequila vinasses.

Keywords: AnSBBR; H2 production; competitive exclusion; dark fermentation; principal component analysis; prokaryotic interactions; tequila-manufacturing residues.