Evaluation of the Antibacterial Material Production in the Fermentation of Bacillus amyloliquefaciens-9 from Whitespotted Bamboo Shark (Chiloscyllium plagiosum).

Bacillus amyloliquefaciens-9 (GBacillus-9), which is isolated from the intestinal tract of the white-spotted bamboo shark (Chiloscyllium plagiosum), can secrete potential antibacterial materials, such as ß-1,3-1,4-glucanase and some antimicrobial peptides. However, the low fermentation production has hindered the development of GBacillus-9 as biological additives. In this study, the Plackett-Burman design and response surface methodology were used to optimize the fermentation conditions in a shake flask to obtain a higher yield and antibacterial activity of GBacillus-9. On the basis of the data from medium screening, M9 medium was selected as the basic medium for fermentation. The data from the single-factor experiment showed that sucrose had the highest antibacterial activity among the 10 carbon sources. The Plackett-Burman design identified sucrose, NH4Cl, and MgSO4 as the major variables altering antibacterial activity. The optimal concentrations of these compounds to enhance antibacterial activity were assessed using the central composite design. Data showed that sucrose, NH4Cl, and MgSO4 had the highest antibacterial activities at concentrations of 64.8, 1.84, and 0.08 g L-1, respectively. The data also showed that the optimal fermentation conditions for the antibacterial material production of GBacillus-9 were as follows: Inoculum volume of 5%, initial pH of 7.0, temperature of 36 °C, rotating speed of 180 rpm, and fermentation time of 10 h. The optimal fermentation medium and conditions achieved to improve the yield of antibacterial materials for GBacillus-9 can enhance the process of developing biological additives derived from GBacillus-9.

Bacillus Amyloliquefaciens-9 as an Alternative Approach to Cure Diarrhea in Saanen Kids.

Bacillus amyloliquefaciens-9 (GBacillus-9), derived from the intestinal tract of the white-spotted bamboo shark, secretes a variety of antimicrobial compounds that inhibit the growth of pathogenic bacteria. In this study, the role of GBacillus-9 in the prevention and treatment of Saanen kids with diarrhea was assessed. Six healthy kids (HL) and six kids with diarrhea (DL) were selected. All kids were fed with 0.3% (w/v) GBacillus-9 (spray power) in raw milk for two weeks. The proportion of kids with diarrhea decreased gradually as the trial progressed, and 100% DL kids were cured at day 15. GBacillus-9 increased the serum immunoglobulin (Ig) G, interleukin (IL)-4, and IL-6 concentration (p < 0.05). The amplicon sequencing analysis of the fecal bacterial community revealed that the fecal microbiota was remarkably different between the HL and the DL groups at day 0. After two weeks of feeding with GBacillus-9, no significant difference in fecal microbiota was observed between HL and DL groups at the phylum level. GBacillus-9 restored the intestinal microbial disorder associated with serum immunoglobulin and interleukin concentration. Correlation analysis showed that GBacillus-9 altered globulin and interleukin concentration and that immunoglobulin was associated with Firmicutes. Collectively, our results revealed that GBacillus-9 improved the gut health of kids by improving microbial homeostasis.

A glucogalactomanan polysaccharide isolated from Agaricus bisporus causes an inflammatory response via the ERK/MAPK and IκB/NFκB pathways in macrophages.

In the present study, a glucogalactomanan polysaccharide isolated from Agaricus bisporus named as TJ3 was investigated its immunomodulatory effects and molecular mechanisms in RAW 264.7 cells. Functional analysis showed TJ3 could inhibit the proliferation of RAW 264.7 cells in a certain concentration range. Moreover, TJ3 treatment increased the expression of inducible nitric oxide synthase (iNOS), which was responsible for increasing nitric oxide (NO) production. TJ3 also increased the expression of cyclooxygenase-2 (COX-2) and the mRNA levels of interleukin 1ß (IL-1ß), interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α) and COX-2 in RAW 264.7 cells. In addition, further elucidation of molecular mechanisms showed that ERK/MAPK and IκB/NFκB pathways were activated by TJ3 treatment via increased phosphorylation levels of ERK1/2 and IκB-α, respectively. These findings indicated that TJ3 had a huge promising immunostimulatory activity and its induced immune responses probably through stimulating the ERK/MAPK and IκB/NFκB pathways. Our results demonstrate that TJ3 may be used as a potent immune modulator.

Identification and Quantification of Triterpenoids in Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), with HPLC-MS/MS Methods.

Ganoderma lucidum is one of the most commonly used mushrooms in traditional Chinese medicine, with significant immunomodulatory and antitumor effects. Triterpenoids are deemed to be the main bioactive components in G. lucidum. In this study, high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (MS) and high-performance liquid chromatography-MS/MS methods were used to identify and quantify, respectively, the main triterpenoids in G. lucidum obtained from different sources. The full MS scan with a daughter ion scan was used to identify a variety of potential derivatives of triterpenoids in the mushroom. Multiple reaction monitoring was performed with transitions of m/z-515.2 → 285.1 (quantifier) and 515.2 → 497.2 (qualifier)-to determine the amount of ganoderic acid A. The developed methods were successfully used to analyze 8 G. lucidum samples from the wild and from cultivated and commercial sources. The results show that the content and composition of triterpenoids differed significantly among these samples. A total of 29 triterpenoids were explicitly or tentatively identified in the wild sample, whereas 23 triterpenoids were found in the cultured sample and 17 in the commercial sample. The results of this study provide a foundation for further research on chemical constituents of, identification of metabolites in, and quality control of the G. lucidum mushroom.