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Moringa oleifera  Lam. (MO) is a fast-growing multi-purpose deciduous tree with high biomass and nutritional value. However, the presence of antinutritional factors, poor palatability, and indigestibility of  Moringa oleifera  leaf meal (MOLM) restrict its application to animal feed. This study aimed to obtain high-quality protein feeds via solid-state fermentation (SSF) of MOLM. The process conditions for increasing the true protein (TP) content using  Aspergillus niger ,  Candida utilis  and  Bacillus subtilis  co-cultures were optimized, and the chemical composition of MOLM was compared before and after fermentation. The results of this study showed that the highest TP content could be obtained through mixed-strain culture of  A. niger ,  C. utilis  and  B. subtilis  at a ratio of 1:1:2. The MOLM was inoculated with  A. niger , followed by  C. utilis  and  B. subtilis  24 h later. The optimized co-culture parameters were as follows: total inoculation size, 24%; temperature, 32 °C; fermentation time, 6.5 days; and initial water content, 60%. The maximum TP yield was 28.37%. Notably, in the fermented MOLM (FMOLM), the content of nutrients such as crude protein (CP), small peptides, and total amino acids (AAs) were significantly increased relative to unfermented MOLM, whereas the contents of crude fiber (CF), tannin, and phytic acid were significantly decreased. MOLM analysis using scanning electron microscopy (SEM) revealed that SSF disrupted the surface structure of MOLM, and sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) indicated that macromolecular proteins were degraded. The in vitro protein digestibility (IVPD) of FMOLM was also improved significantly. Our findings suggest that multi-strain fermentation with  A. niger ,  C. utilis  and  B. subtilis  improves the nutritional quality of MOLM, rendering it a viable functional feedstuff for use in livestock industries in the future.

Solid state fermentation of Moringa oleifera leaf meal by mixed strains for the protein enrichment and the improvement of nutritional value