Phenotypic plasticity in the productions of virulence factors within and among serotypes in the Cryptococcus neoformans species complex

The Cryptococcus neoformans species complex (CNSC) is a common opportunistic human fungal pathogen and the most frequent cause of fungal meningitis. There are three major serotypes in CNSC: A, D, and their hybrids AD, and they have different geographic distributions and medical significance. Melanin pigment and a polysaccharide capsule are the two major virulence factors in CNSC. However, the relationships between serotype and virulence factor production and how environmental factors might impact their relationships are not known. This study investigated the expressions of melanin and capsular polysaccharide in a genetically diverse group of CNSC strains and how their phenotypic expressions were influenced by oxidative and nitrosative stress levels. We found significant differences in melanin and capsular polysaccharide productions among serotypes and across stress conditions. Under oxidative stress, the laboratory hybrids exhibited the highest phenotypic plasticity for melanin production while serotype A showed the highest for capsular polysaccharide production. In contrast, serotype D exhibited the highest phenotypic plasticity for capsular polysaccharide production and clinical serotype AD the highest phenotypic plasticity for melanin production under nitrosative stress. These results demonstrated that different serotypes have different environmental condition-specific mechanisms to modulate the expression of virulence factors.

ARTP Mutagenesis to Improve Mycelial Polysaccharide Production of Grifola frondosa Using a Mixture of Wheat Bran and Rice Bran as Substrate

Mycelial polysaccharides from Grifola frondosa have shown potential for the prevention of chronic diseases. Atmospheric and room temperature plasma (ARTP) technology was used to enhance the ability of G. frondosa to efficiently utilize a mixture of rice bran and wheat bran in the production of mycelial polysaccharides. The ARTP-mutant G. frondosa GFA2 had an improved growth rate of 6.0 mm/d and polysaccharide yield of 2.65 g/L and showed stable genetic characteristics. Uniform design experiments showed that polysaccharide yield could be increased to 5.90 g/L using the optimized conditions of 10.0 g/L rice bran and 110.0 g/L wheat bran while omitting KH2PO4 and MgSO4·7H2O. Gas chromatography demonstrated that GFA2 polysaccharides were composed of the monosaccharides rhamnose, arabinose, fucose, xylose, mannose, glucose, and galactose. This study provides an effective strategy for improving polysaccharide production in edible fungi while proposing the added-value utilization of rice and wheat brans.

Effect of Alkali-Treated Birch Sawdust on the Lignocellulase Secretion and Exo-Polysaccharide Production by Inonotus Obliquus Under Submerged Fermentation and its Lignocellulose Degradation Patterns

The objectives of this study were to investigate the medicinal mushroom Inonotus obliquus on the production of polysaccharides and changes of extracellular lignocellulolytic enzymes during submerged fermentation using alkali-treated birch sawdust as substrate. Meanwhile, in order to explore the degradation mode of lignocellulose in alkali-treated birch sawdust, degradation analysis of three components of lignocellulose was carried out. The fungus process in alkali-treated birch sawdust medium resulted in a higher degradation rate of cellulose, hemicellulose, and lignin of 39.24%, 51.00% and 31.3% after 11 days of submerged fermentation by the mycelium of I. obliquus, respectively. Maximal polysaccharide production and α-glucosidase inhibition rate determined in the alkali-treated birch sawdust medium were 6.93 mg/mL and 55.80%,while they were 4.98 mg/mL and 27.89% in the control. Moreover, high activities of Laccase (51.95 IU/mL) , CMCase (1.35 IU/mL) , FPA (0.50 IU/mL) and β-glucosidase (0.55 IU/mL) were observed in alkali-treated birch sawdust medium, respectively. The results demonstrated that the addition of alkali-treated birch sawdust could promote the yield of active polysaccharides and induce the production of cellulase and xylanase, indicating that alkali pretreatment was conducive to utilization of birch sawdust by I. obliquus.

Effects of nitrogen from different sources on mycelial biomass and polysaccharide production and pellet morphology in submerged cultures of Grifola frondosa

The effects of nitrogen in the medium on the production of mycelial biomass, extracellular polysaccharides (EPS), and intracellular polysaccharides (IPS) was investigated in submerged cultures of Grifola frondosa. In addition, the effects on pellet morphology were examined. The maximum production levels of mycelial biomass (2.32 g/L), EPS (1.58 g/L), and IPS (29.1 mg/L) were obtained when the nitrogen sources in the medium were yeast extract, malt extract, and peptone, respectively. Using yeast extract as the nitrogen source yielded the maximum mycelial biomass, and morphological characterization revealed a composition of 47% large pellets (fraction L), 20% small pellets (fraction S), and 33% adhesive mycelia (fraction A). The maximum circularity value and the minimum roughness value of the pellets were observed using yeast extract cultures. Both the compactness (0.53) and circularity (0.15) of the pellets were the lowest among the seven types of nitrogen sources, but the roughness (2.86) was the highest in malt extract, which was the nitrogen source that resulted in maximum polysaccharide production. The results revealed that the production levels of mycelial biomass, EPS, and IPS of G. frondosa were associated with changes in pellet morphology due to the source of nitrogen in the medium.

Influence of sub-inhibitory concentrations of antimicrobials on micrococcal nuclease and biofilm formation in Staphylococcus aureus

A major contributor to biomaterial associated infection (BAI) is Staphylococcus aureus. This pathogen produces a protective biofilm, making eradication difficult. Biofilms are composed of bacteria encapsulated in a matrix of extracellular polymeric substances (EPS) comprising polysaccharides, proteins and extracellular DNA (eDNA). S. aureus also produces micrococcal nuclease (MN), an endonuclease which contributes to biofilm composition and dispersion, mainly expressed by nuc1. MN expression can be modulated by sub-minimum inhibitory concentrations of antimicrobials. We investigated the relation between the biofilm and MN expression and the impact of the application of antimicrobial pressure on this relation. Planktonic and biofilm cultures of three S. aureus strains, including a nuc1 deficient strain, were cultured under antimicrobial pressure. Results indicated that nuc1 deletion stimulates the polysaccharide production per CFU in the biofilm in in vitro biofilms. Also antimicrobial pressure of ciproflocacin, doxycycline and erythromycin resulted in significantly increased quantities of polysaccharides per CFU, but this did only coincide with significantly reduced MN activity in erythromycin. Resveratrol significantly reduced MN production per CFU but did not affect polysaccharides production. In conclusion, various antimicrobials impact the balance of eDNA, polysaccharides and MN production, all in a different way.