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To investigate the structural and functional similarities of microbial communities in burnt-sweetness alcoholized tobacco as a function of distance from the equator and their effects on tobacco quality, we sampled alcoholized tobacco from Chenzhou, Hunan Province, China and from Brazil and Zimbabwe, which are also burnt-sweetness-type tobacco producing regions, and performed high-throughput sequencing of tobacco bacterial and fungal communities along with an analysis of the main chemical constituents of the tobacco to analyze differences in the quality of the tobacco and similarities in the structure of the microbial communities. The total nitrogen, nicotine and starch contents of Chenzhou tobacco were greater than those of Brazilian and Zimbabwean tobacco, and the total sugar and reducing sugar contents of the Brazilian and Zimbabwean tobacco were greater than those of the Chenzhou tobacco (P < 0.05). The alpha diversity indices of the bacterial communities in Chenzhou tobacco were lower than those in the Brazilian and Zimbabwean tobacco, and the alpha diversity indices of the fungal communities in Chenzhou tobacco were greater than those in the Brazilian and Zimbabwean tobacco (P < 0.05). In the ecological networks, bacterial–fungal interactions in the Brazilian and Zimbabwean tobacco were more complex than those in the Chenzhou tobacco, and the microbial ecological networks of the burnt-sweetness-type tobacco from three different regions were dominated by competitive relationships. The microbial community composition of Chenzhou tobacco was similar to that of Brazilian tobacco at the bacterial genus and fungal phylum level, with Sphingomonas being a significantly enriched genus in Brazilian tobacco and a key genus in the Chenzhou network that is able to participate in the degradation of polyphenols and aromatic compounds. Functional microbes related to aromatic compounds and cellulose degradation were significantly more abundant in the Brazilian and Zimbabwean tobacco than in Chenzhou tobacco, and the related degradation of tobacco substances was responsible for the better quality of the Brazilian and Zimbabwean tobacco. In conclusion, there are similarities in the structure, composition and functional flora of microbial communities in tobacco from Chenzhou and Brazil because these regions have similar latitudinal distributions. This study provides theoretical support for selecting cultivation regions for the burnt-sweetness-type alcoholized tobacco and for the alcoholization of tobacco leaves.
To investigate the structural and functional similarities of microbial communities in burnt-sweetness alcoholized tobacco as a function of distance from the equator and their effects on tobacco quality, we sampled alcoholized tobacco from Chenzhou, Hunan Province, China and from Brazil and Zimbabwe, which are also burnt-sweetness-type tobacco producing regions, and performed high-throughput sequencing of tobacco bacterial and fungal communities along with an analysis of the main chemical constituents of the tobacco to analyze differences in the quality of the tobacco and similarities in the structure of the microbial communities. The total nitrogen, nicotine and starch contents of Chenzhou tobacco were greater than those of Brazilian and Zimbabwean tobacco, and the total sugar and reducing sugar contents of the Brazilian and Zimbabwean tobacco were greater than those of the Chenzhou tobacco (P < 0.05). The alpha diversity indices of the bacterial communities in Chenzhou tobacco were lower than those in the Brazilian and Zimbabwean tobacco, and the alpha diversity indices of the fungal communities in Chenzhou tobacco were greater than those in the Brazilian and Zimbabwean tobacco (P < 0.05). In the ecological networks, bacterial–fungal interactions in the Brazilian and Zimbabwean tobacco were more complex than those in the Chenzhou tobacco, and the microbial ecological networks of the burnt-sweetness-type tobacco from three different regions were dominated by competitive relationships. The microbial community composition of Chenzhou tobacco was similar to that of Brazilian tobacco at the bacterial genus and fungal phylum level, with Sphingomonas being a significantly enriched genus in Brazilian tobacco and a key genus in the Chenzhou network that is able to participate in the degradation of polyphenols and aromatic compounds. Functional microbes related to aromatic compounds and cellulose degradation were significantly more abundant in the Brazilian and Zimbabwean tobacco than in Chenzhou tobacco, and the related degradation of tobacco substances was responsible for the better quality of the Brazilian and Zimbabwean tobacco. In conclusion, there are similarities in the structure, composition and functional flora of microbial communities in tobacco from Chenzhou and Brazil because these regions have similar latitudinal distributions. This study provides theoretical support for selecting cultivation regions for the burnt-sweetness-type alcoholized tobacco and for the alcoholization of tobacco leaves.
Dairy mastitis is one of the most common diseases in dairy farming, and the formation of pathogenic bacteria biofilms may be an important reason why traditional antibiotic therapy fails to resolve some cases of dairy mastitis. We isolated and identified three strains of A. lwoffii were with strong biofilm forming ability from dairy cow mastitis samples from Chongqing dairy farms in China. In order to investigate the effect of novel anti-biofilm peptide CRAMP-34 on A.lwoffii biofilms, the anti-biofilm effect was evaluated by crystal violet staining, biofilms viable bacteria counting and confocal laser scanning microscopy (CLSM). In addition, transcriptome sequencing analysis, qRT-PCR and phenotypic verification were used to explore the mechanism of its action. The results showed that CRAMP-34 had a dose-dependent eradicating effect on A. lwoffii biofilms. Transcriptome sequencing analysis showed that 36 differentially expressed genes (11 up-regulated and 25 down-regulated) were detected after the intervention with the sub-inhibitory concentration of CRAMP-34. These differentially expressed genes may be related to enzyme synthesis, fimbriae, iron uptake system, capsular polysaccharide and other virulence factors through the functional analysis of differential genes. The results of subsequent bacterial motility and adhesion tests showed that the motility of A.lwoffii were enhanced after the intervention of CRAMP-34, but there was no significant change in adhesion. It was speculated that CRAMP-34 may promote the dispersion of biofilm bacteria by enhancing the motility of biofilm bacteria, thereby achieving the effect of eradicating biofilms. Therefore, these results, along with our other previous findings, suggest that CRAMP-34 holds promise as a new biofilm eradicator and deserves further research and development.
ObjectiveThis study aims to investigate the effects of pectin-degrading bacteria on the microbial community and physicochemical properties during the fermentation process of cigar tobacco, evaluating its potential in reducing green bitterness and enhancing aroma.MethodsBy isolating and screening pectin-degrading bacteria, high-throughput sequencing and physicochemical analysis were employed to compare the microbial flora and physicochemical component differences in different treatment groups of cigar tobacco. Furthermore, correlation analysis was performed to examine the relationships between these variables.ResultsThe results showed that the strains YX-2 and DM-3, isolated from the cigar tobacco variety “Yunxue No. 1,” exhibited strong pectin-degrading abilities. Phylogenetic analysis revealed that strain YX-2 is highly homologous to Bacillus flexus, while strain DM-3 is highly homologous to Bacillus siamensis. After fermentation, the addition of strains YX-2 and DM-3 significantly reduced the pectin content in the tobacco leaves, increased the total sugar and reducing sugar content, reduced green bitterness, and markedly enhanced the total aroma components. Notably, DM-3 exhibited outstanding performance in the production of Maillard reaction products. Microbial community analysis showed that the addition of pectin-degrading bacteria significantly increased the diversity of both bacteria and fungi, especially in the TDM3 group, where the relative abundance of Pseudomonas was notably elevated. Correlation analysis revealed that Pseudomonas had a significant positive correlation with both reducing sugar and total sugar, and a significant negative correlation with pectin, indicating its important role in sugar metabolism and pectin degradation. Additionally, fungal genera such as Cercospora were significantly negatively correlated with total sugar and total nitrogen, while Eurotium was closely associated with pectin degradation and reducing sugar accumulation.ConclusionThis study found that the addition of Pectin-degrading bacteria YX-2 and DM-3 significantly optimized the microbial community structure during the cigar tobacco fermentation process and improved the physicochemical properties of the tobacco leaves, with notable effects in reducing green bitterness and enhancing aroma.
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