Intestinal mucosal bacterial diversity of antibiotic-associated diarrhea (AAD) mice treated with Debaryomyces hansenii and Qiweibaizhu powder.

The aim was to investigate the combined effect of Debaryomyces hansenii and Qiweibaizhu powder (QWBZP) on the bacterial diversity of the intestinal mucosa of antibiotic-associated diarrhea (AAD) mice, for the potential treatment of diarrhea, especially which is induced by administration of antibiotics. Eighteen (18) mice were randomly assigned to three equal groups of six mice, namely Normal (mn group), Placebo control (mm group) and D. hansenii and QWBZP (DQ) treatment (mdq group). Mice were gavaged with a solution (23.33 mL·kg-1·day-1) consisting of gentamicin and cefradine to establish AAD. The DQ treatment group was gavaged with DQ for 4 days, and sterile water was used as a placebo control. The metagenome DNA of the intestinal mucosal microbiota was extracted, and the 16S rRNA gene was sequenced. Analysis showed that there were 288 OTUs for the normal group, 443 for the placebo control group, and 229 for the DQ treatment group. Phylogenetically, the gut microbiota of the DQ treatment group and the normal group were closer to each other than to the placebo control group. Both the DQ and placebo-treated groups included Stenotrophomonas, Robinsoniella, Bacteroidales S24-7 group norank, Citrobacter, and Glutamicibacter, but their abundances were significantly higher in the DQ treatment group than in the placebo control group. This suggested that the combined use of D. hansenii and QWBZP overcame the influence of dysbacteriosis and could lead to the recovery of intestinal mucosal microbiota homeostasis. This positive effect is likely related to short-chain fatty acid (SCFA)-producing bacteria, such as members of Micrococcaceae, Lachnospiraceae, and Bacteroidales S24-7 group, which could play beneficial roles in protecting the mucosal barrier and stimulating the immune response in mice.

Effect of Debaryomyces hansenii combined with Qiweibaizhu powder extract on the gut microbiota of antibiotic-treated mice with diarrhea.

The aim of this study was to investigate the effects of an extract of Qiweibaizhu powder combined with Debaryomyces hansenii on the gut microbiota of antibiotic-treated mice with diarrhea. Mice were gavaged with a mixture of gentamycin sulfate and cefradine to induce diarrhea. After diarrhea was observed, 25% dose of ultra-micro Qiweibaizhu powder extract combined with 25% dose of Debaryomyces hansenii (QCD) was gavaged to mice with diarrhea. DNA of intestinal contents in mice was extracted for 16S rRNA gene sequence analysis by high-throughput sequencing following treatment finished. The results showed that the QCD increased the species richness and diversity, but did not recover the diversity to the original level. Antibiotics and QCD significantly altered the composition of gut microbiota at different taxonomic levels. At the genus level, the relative abundance of Bacteroidales S24-7 group_unidentified and Bacteroides returned to baseline after QCD treatment. Additionally, QCD suppressed the growth of Oscillospira and Ruminococcus, and promoted the proliferation of Erysipelotrichaceae_norank and Blautia compared with the healthy and diarrheal mice. Our results indicated that QCD modulated the diversity and composition of the gut microbiota in antibiotic-treated mice with diarrhea. The synergistic effect between Qiweibaizhu powder extract and Debaryomyces hansenii may be related to Bifidobacterium and Bacteroidales S24-7 group_unidentified.

Bacterial Lactase Gene Characteristics in Intestinal Contents of Antibiotic-Associated Diarrhea Mice Treated with Debaryomyces hansenii.

BACKGROUND Debaryomyces hansenii exhibits a therapeutic effect on antibiotic-associated diarrhea (AAD) by promoting the growth of beneficial intestinal bacteria. Previous research has reported that AAD involves not only dysbacteriosis but also dysfunction of the activity of intestinal enzymes (such as lactase). Enzyme activities can be influenced by many other factors, such as gene expression. The present study showed that D. hansenii has a curative effect on AAD at the lactase gene level. MATERIAL AND METHODS The effect of D. hansenii on the lactase gene from intestinal bacteria in AAD mice was investigated. The diarrhea model was established with a gentamycin sulfate and cefradine capsule mixture. The antibiotic mixture (23.33 mL·kg⁻¹·day⁻¹) was intragastrically administered for 5 days. Subsequently, half of the diarrhea mice were treated with D. hansenii twice a day for 3 days while the other mice were intragastrically administered with the same volume of distilled water. Next, the intestinal contents were collected, and metagenomic DNA was extracted for high-throughput sequencing analysis. RESULTS The Chao1 and Shannon indices decreased significantly following treatment with D. hansenii (P<0.01 and P<0.05, respectively). Moreover, the clusters in the D. hansenii group mice were quite different from those in the normal group mice and model group mice. Following treatment with D. hansenii, the quantity of lactase genes in Enterobacter sp. 638 and Modestobacter increased markedly, and the richness of intestinal bacterial lactase genes in Fretibacterium recovered. CONCLUSIONS D. hansenii altered the lactase-producing bacterial community structure and promoted the growth of several critical lactase-producing bacteria, such as Enterobacter sp. 638 and Modestobacter.

Influence of Debaryomyces hansenii on bacterial lactase gene diversity in intestinal mucosa of mice with antibiotic-associated diarrhea.

AIM: The current study aimed to investigate the effects of Debaryomyces hansenii on the diversity of bacterial lactase gene in the intestinal mucosa of antibiotic-associated diarrhea (AAD) mice. METHODS: Eighteen mice were randomly divided into three groups (6 mice per group): healthy control group, diarrhea model group and D. hansenii treatment group. The antibiotic-associated diarrhea model was established by intragastric administration with a mixture of cephradine and gentamicin sulfate (23.33 mL·kg-1·d-1) twice a day for 5 days continuously. After establishing the AAD model, the mice in the D. hansenii treatment group were gavaged with D. hansenii for three days, while other groups were gavaged with distilled water. Then, the intestinal mucosa of all three groups was collected and DNA was extracted in an aseptic environment for the following analysis. RESULTS: The difference in the richness and homogeneity of the bacterial lactase gene among all samples were inapparent, as the difference in the Chao1, ACE, Simpson and Shannon indices among the three groups were insignificant (P>0.05). NMDS analysis also showed that the distance of the samples among the three groups was unobvious. Furthermore, the bacterial lactase gene in the mucosa mainly originated from Actinobacteria, Firmicutes and Proteobacteria. Compared with the healthy control group, the abundance of lactase genes originating from Cupriavidus, Lysobacter, Citrobacter, Enterobacter and Pseudomonas was increased in the D. hansenii treatment group, while the lactase gene from Acidovorax and Stenotrophomonas decreased (p < 0.01 or p < 0.05) in the diarrhea model group and the D. hansenii treatment group. CONCLUSION: D. hansenii was capable of improving the growth of some key lactase-producing bacteria like Deinococcus, Cupriavidus and Lysobacter for treating AAD.

Effects of Debaryomyces hansenii treatment on intestinal mucosa microecology in mice with antibiotic-associated diarrhea.

AIM: To confirm the effects of Debaryomyces hansenii on intestinal microecology in mice with antibiotic-associated diarrhea (AAD). METHODS: This study took the mucosal microecology as the entry point and an antibiotic mixture was used to induce diarrhea in mice. D. hansenii suspension was used to treat the mice and the bacterial communities of mucosa was analyzed using high-throughput sequencing. RESULTS: The Shannon-Wiener index indicated that the sequencing depth is reasonable and reflected the majority of microbial information. The principal coordinate analysis results showed that mice in the treatment group and the normal group had a similar microbial community structure, while differences in microbial community structure were observed between the model group and the treatment group. The inter-group bacterial structures were analyzed at the phylum level and genus level. The results revealed that antibiotic treatment increased the proportion of Proteobacteria and decreased the proportion of Bacteroides, while D. hansenii treatment inhibited the increase in Proteobacteria. Linear discriminant analysis coupled with effect size measurements (LEfSe) suggested d that the beneficial bacteria Candidatus Arthromitus were the only common bacteria in the normal group (P<0.05). CONCLUSION: The treatment with D.hansenii could contribute to the maintenance of the structure of the mucosal microbiota in comparison with the normal group and inhibit the proliferation of opportunistic bacteria. However, high-dose antibiotic treatment causes mucosal dysbiosis and the proliferation of opportunistic bacteria during the self-recovery period, such as Pseudoalteromonas, Alteromonas, Vibrio.

Effects of Debaryomyces hansenii treatment on intestinal microorganisms in mice with antibiotics-induced diarrhea.

To investigate the influence of Debaryomyces hansenii treatment on intestinal microorganisms in mice with antibiotics-induced diarrhea, mouse model of antibiotics-induced diarrhea was created by gavaging mice with mixed antibiotics (23.33 mL/kg/days) composed of gentamycin sulfate and cefradine for 5 days. Mice with the symptom of diarrhea were then treated with D. hansenii by intragastric administration. The control group mice were given with sterile water. After 4 day treatment, total DNA of intestinal microflora of treated and control mice was extracted, and their quantities were measured by sequencing the V4 region of 16S rDNA. The results showed that when compared to the control (sterile water), treatment with D. hansenii increased the operational taxonomic units (OTUs) of intestinal bacteria. The Chao index in diarrhea treated group was higher than diarrhea control group and was similar to healthy control group, while all differences had no significance (P > 0.05). D. hansenii treatment increased the Shannon index but not significantly (P > 0.05). Moreover, there was not significant impact on density and diversity of intestinal bacterial population at phylum and genus levels (P > 0.05). Interestingly, D. hansenii treatment recovered the population density of certain bacterium species, such as Bacteroidaceae (in family level) (P < 0.05). Our results indicate that D. hansenii has potency of adjusting the density and diversity of intestinal bacteria and recovering the population density of Bacteroidaceae in family level.