Low fertilizer quality and remediation ability are considered the major factors hampering the land application of anaerobic digestate. Therefore, the role of Myrothecium verrucaria treatment in enhancing the fertilizer quality and remediation ability of digestate for land application was explored. Higher content of humic acid (7.5 g/L) with a higher degree of humification index and oxygen-containing functional groups was observed in the digestate receiving Myrothecium verrucaria treatment. Likewise, humic acid formed from Myrothecium verrucaria treatment had a higher capacity of heavy metal binding. Moreover, the viable and culturable count of S. faecalis, S. typhi, C. perfringens, and E. coli pathogens in the digestate decreased to approximately 12.50%, 41.70%, 18.87%, and 50.00% and 25.97%, 64.44%, 37.51%, and 75.27%, respectively, after treatment with Myrothecium verrucaria. This study provided a novel strategy to enhance the fertilizer quality, remediation ability, and biological safety of anaerobic digestate for land application.

Cryptic species are morphologically indistinguishable but possess genetically distinct taxa. Alternative splicing (AS) regulates physiological processes, thereby facilitating ecological adaptation and evolution. To explore the sex-specific differences in transcriptional regulation among cryptic species, we profiled both AS and gene expression in two cryptic species of Wiebesia pumilae (WPDZ19 and WPHS), which differ in Wolbachia infection status. The results showed that 101 and 71 differentially alternatively spliced genes (DASs) were identified in female and male groups, respectively. Functional enrichment revealed that female DASs were significantly enriched in mitotic cell cycle process, cytoskeleton organization, cellular component organization, and DNA damage. On the other hand, male DASs were predominantly related to actin, cytoskeleton, and muscle development. Gene set enrichment analysis of DASs also revealed that the regulation of mitotic nuclear division and meiotic nuclear division were enriched in female and male groups, respectively. We identified 4509 DEGs in females and 3645 in males, with minimal overlap between DASs and DEGs. Moreover, RT-PCR has been used to validate the key genes. Our results revealed sexually divergent transcriptional regulation patterns between WPDZ19 and WPHS, suggesting a potential association with Wolbachia persistence. Our findings provide new insights into the study of adaptive evolution of cryptic species.