Cow manure (CM) generation in large volumes has for long been considered a waste management challenge. However, the organic content of CM signals opportunities for the production of value-added bioproducts such as volatile fatty acids (VFAs) through anaerobic digestion (AD). However, a robust VFAs fermentation process requires effective methane formation inhibition and enhance VFAs recovery. In this study, thermal pretreatment was applied to inhibit methanogens for enhanced VFAs production and an immersed membrane bioreactor (iMBR) for in situ recovery of VFAs in a semi-continuous AD. Maximal VFAs yield of 0.41 g VFAs/g volatile solids (VS) was obtained from thermally-treated CM without inoculum addition. The CM was further fed to the iMBR operating at organic loading rates of 0.8–4.7 gVS/L.d. The VFAs concentration increased to 6.93 g/L by rising substrate loading to 4.7 g VS/L.d. The applied iMBR set-up was successfully used for stable long-term (114 days) VFAs production and recovery.

Acidogenic fermentation of chicken manure (CM) for production and recovery of volatile fatty acids (VFA) is an interesting biological waste-to-value approach compared to benchmark organic waste management strategies. Considering the wide range of high value applications of VFA, a semi-continuous immersed anaerobic membrane bioreactor (AnMBR) was applied to boost VFA productivity and yield, while reducing downstream processing stages assisting the recovery of VFA. In this regard, the effect of parameters such as pH and organic loading rates (OLR) on the overall bioconversion and filtration performance was investigated. Thermal-shocked CM was applied both as inoculum and substrate. A very high VFA yield (0.90 g-VFA/g-VS) was obtained in the treatment with no pH control (~8.2) at an OLR of 2 g-VS/(L·d), presenting 24% higher yield compared to that of the controlled pH. Batch assays further demonstrated the enhanced hydrolysis and acidogenesis activities at weak alkaline conditions. A long-term (78 days) fermentation and filtration was successfully performed, where stable membrane filtration performance was experienced for about 50 days under high-solid (suspended solid of 37–45 g/L) and high flux (20 L/(m2·h)) conditions. Results suggest that AnMBR of CM is a feasible and promising process for VFA production and recovery.

In a circular economy approach, edible filamentous fungi (single cell protein) can be cultivated on volatile fatty acids (VFAs) derived from anaerobic digestion (AD) of organic-rich waste streams. In this study, the effect of pH, concentration/distribution of VFAs, nutrient supplementation, and type of waste on Aspergillus oryzae cultivation on synthetic VFAs, and actual VFAs derived from AD of food waste and cow manure were investigated. The optimal pH for A. oryzae growth on VFAs were 6 and 7 with maximum acetic acid consumption rates of 0.09 g/L. h. The fungus could thrive on high concentrations of acetic (up to 9 g/L) yielding 0.29 g dry biomass/gVFAs(fed). In mixed VFAs cultures, A. oryzae primarily consumed caproic and acetic acids reaching a biomass yield of 0.26 g dry biomass/gVFAs(fed) (containing up to 41% protein). For waste-derived VFAs at pH 6, the fungus successfully consumed 81-100% of caproic, acetic, and butyric acids.

Considering the projected demand for protein supplementation in animal feed, as well as prioritizing plant-based protein provision for the growing human population, great stress is imposed on conventional protein sources, calling for new sustainable alternatives. In this regard, the production and application of single-cell proteins (SCPs) has proven to be a promising alternative. Therefore, in this study, volatile fatty acids (VFAs) effluents recovered from anaerobically digested FW, CKM, CM, and their combinations were applied for the cultivation of edible filamentous fungi Aspergillus oryzae. The biomass was further evaluated considering its protein, fat and alkali insoluble material contents. The maximum fungal biomass yielded of 0.47 ± 0.00 and 0.37 ± 0.00 g dry biomass/g tVFAsCODeq.consumed, with up to 47% protein and 5% fat content successfully cultivated in shake flasks and bench scale reactors, respectively. In addition to the production of protein-rich biomass, significant reductions in medium COD (25–58%) and ammonium (33–48%) were achieved. The results presented in this research work imply that using waste-derived VFAs for the production of animal feed grade SCP is an innovative approach that can contribute to the economy and sustainability of animal feed production process. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Organic-waste-derived volatile fatty acids (VFAs) are promising substrates for fungal biomass cultivation, offering a nutrient-rich medium capable of meeting microbial growth requirements. However, the growth and biomass productivity are highly influenced by the VFAs’ composition and mode of operation. This study investigated the cultivation of Aspergillus oryzae fungal biomass using agro-industrial-derived VFA effluent, employing repeated-batch and fed-batch (stepwise and continuous-feeding) cultivation modes to evaluate fungal growth and biomass composition. The highest dry biomass yield of 0.41 dry biomass/gVFAs_fed (g/g) was achieved in fed-batch mode with continuous feeding, where the biomass exhibited pellet morphology, facilitating ease of harvesting. The crude protein content varied according to the cultivation strategy, reaching 45–53% in continuous-feeding fed-batch mode, while it was 34–42% in stepwise fed-batch mode. Additionally, the fungal biomass contained significant levels of essential macronutrients and trace elements, including Mg, Ca, K, Mn, and Fe, which are crucial if the biomass is intended to be used in animal feed formulations. This study highlights the effects of cultivation modes on biomass composition and the potential of VFA-derived fungal biomass as a sustainable feed ingredient.

The resource-intensive nature of the ruminant farming sector, which has been exacerbated by population growth and increasing pressure to reduce feed antibiotics and growth promoters, has sparked interest in looking for sustainable alternative feed sources to enhance ruminant production efficiency. Edible filamentous fungi, rich in macronutrients like proteins, offer promise in reducing the reliance on conventional protein sources and antimicrobials to improve feed quality and animal performance. The inclusion of single-cell proteins, particularly filamentous fungi, in ruminant feed has long been of scientific and industrial interest. This review focuses on the potential application of the extensively studied Aspergillus oryzae and its fermentation extracts in ruminant nutrition. It provides an overview of conventional ruminant feed ingredients, supplements, and efficiency. Additionally, this review analyzes the re-utilization of organic residues for A. oryzae cultivation and examines the effects of adding fungal extracts to ruminant feed on ruminal digestibility and animal performance, all within a circular bioeconomy framework.

Background

As demand for high quality animal feed continues to raise, it becomes increasingly important to minimize the environmental impact of feed production. An appealing sustainable approach to provide feed fractions is to use organic residues from agro-food industry. In this regard, volatile fatty acids (VFAs) such as acetic, propionic and butyric acids, derived from bioconversion of organic residues can be used as precursors for production of microbial protein with ruminant feed inclusion potential. This study aims to investigate the in vitro digestibility of the Aspergillus oryzae edible fungal biomass cultivated on VFAs-derived from anaerobic digestion of residues. The produced fungal protein biomass, along with hay clover silage and rapeseed meal were subjected to various in vitro assays using two-stage Tilley and Terry (TT), gas, and bag methods to evaluate and compare its digestibility for application in ruminant feed.

Results

The produced fungal biomass contained a higher crude protein (CP) (41%–49%) and rather similar neutral detergent fiber (NDF) (41%–56%) compared to rapeseed meal. The rumen in vitro dry matter digestibility (IVDMD) of the fungal biomass in the TT method ranged from 82% to 88% (statistically similar to that of the gas method (72% to 85%)). The IVDMD of fungal biomass were up to 26% and 40% greater than that of hay clover silage and rapeseed meal, respectively. The type of substrate and bag method had pronounced effect on the fermentation products (ammonium-N (NH4+-N), total gas and VFAs). Fungal biomass digestion resulted in the highest release of NH4+-N (340–540 mg/L) and the ratio of acetate to propionate ratio (3.5) among subjected substrates.

Conclusion

The results indicate that gas method can be used as a reliable predictor for IVDMD as well as fermentation products. Furthermore, the high IVDMD and fermentation product observed for Aspergillus oryzae fungal biomass digestion, suggest that the supplementation of fungal biomass will contribute to improving the rumen digestion by providing necessary nitrogen and energy to the ruminant and microbiota.