The aim of this paper is to describe a study of the anaerobic digestion of industrial citrus solid waste (ISCW) in both batch and semi-continuous modes for the production of bioenergy without the elimination of D-limonene. The study was conducted at the pilot plant level in an anaerobic reactor with a working volume of 220 L under mesophilic conditions of 35 ± 2 °C. Cattle manure (CM) was used as the inoculum. Three batches were studied. The first batch had a CM/ISCW ratio of 90/10, and Batches 2 and 3 had CM/ISCW ratios of 80/20 and 70/30, respectively. In the semi-continuous mode an OLR of approximately 8 g total chemical oxygen demand (COD)/Ld (4.43 gVS/Ld) was used. The results showed that 49%, 44%, and 60% of volatile solids were removed in the batch mode, and 35% was removed in the semi-continuous mode. In the batch mode, 0.322, 0.382, and 0.316 LCH₄ were obtained at STP/gVS_removed. A total of 24.4 L/d (34% methane) was measured in the semi-continuous mode. Bioenergy potentials of 3.97, 5.66, and 8.79 kWh were obtained for the respective batches, and 0.09 kWh was calculated in the semi-continuous mode. The citrus industry could produce 37 GWh per season. A ton of processed oranges has a bioenergy potential of 162 kWh, which is equivalent to 49 kWh of available electricity ($3.90).

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 an anaerobic sequencing batch biofilm reactor (AnSBBR) fed with tequila vinasses in co-digestion with acid hydrolysates of Agave tequilana var. azul bagasse (ATAB). Methods and Results The AnSBBR was operated for H₂ production to correlate changes in physicochemical and biological variables by Principal Components 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 of 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.