An amino acid analyzer method for the simultaneous determination of 20 free amino acids (FAAs) and glutathione (GSH) in Penaeus vannamei (PV), Penaeus vannamei, Penaeus hidulis (PH) and Penaeus japonicus (PJ) were developed. The effects of different concentrations of trichloroacetic acid (TCA) and ethanol on the extraction of free amino acids were investigated, and 120 g·L⁻¹ TCA was found to be ideal. The target analytes were eluted in sodium citrate buffer B1 (pH = 3.3) containing 135 mL·L⁻¹ ethanol and 1 mol·L⁻¹ sodium hydroxide (7 mL) and at the optimizing conversion time of sodium citrate buffer B2 (pH = 3.2) and sodium citrate buffer B3 (pH = 4.0) of 5.6 min, and the effective separation was achieved within 29.5 min. The developed method showed good linearity (R² ≥ 0.9991) in the range of 1–250 µg·mL⁻¹ with good intra-day and inter-day precision (relative standard deviations ≤ 2.38%) and spike recovery (86.42–103.64%). GSH and cysteine were used to identify marine prawn and freshwater shrimp. Hydroxyproline and serine were used to distinguish PV and Macrobrachium nipponense (MN) from others, respectively. The highest content of the total FAAs was found in PV, and principal component analysis revealed that PV had the highest comprehensive score for FAAs and GSH. Arginine was found to have the greatest influence on shrimp flavor.

The present study investigated whether the purified polysaccharide from Cereus sinensis (CSP-1) had beneficial effects on mice with antibiotic-associated diarrhea (AAD). The effects of CSP-1 on gut microbiota were evaluated by 16S rRNA high-throughput sequencing. Results showed that CSP-1 increased the diversity and richness of gut microbiota. CSP-1 enriched Phasecolarctobacterium, Bifidobacterium and reduced the abundance of Parabacteroides, Sutterella, Coprobacillus to near normal levels, modifying the gut microbial community. Microbial metabolites were further analyzed by gas chromatography-mass spectrometry (GC-MS). Results indicated CSP-1 promoted the production of various short-chain fatty acids (SCFAs) and significantly improved intestinal microflora dysfunction in AAD mice. In addition, enzyme linked immunosorbent assay and hematoxylin-eosin staining were used to assess the effects of CSP-1 on cytokine levels and intestinal tissue in AAD mice. Results demonstrated that CSP-1 inhibited the secretion of interleukin-2 (IL-2), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) and improved the intestinal barrier. Correspondingly, the daily records also showed that CSP-1 promoted recovery of diarrhea status score, water intake and body weight in mice with AAD. In short, CSP-1 helped alleviate AAD by regulating the inflammatory cytokines, altering the composition and richness of intestinal flora, promoting the production of SCFAs, improving the intestinal barrier as well as reversing the dysregulated microbiota function.