Multidrug-resistant bacteria, including carbapenem-resistant Klebsiella pneumoniae (CRKP), are becoming an increasing health crisis worldwide. For CRKP, colistin is regarded as "the last treatment option." In this study, we isolated a clinical CRKP strain named as K. pneumoniae R10-341. Phenotyping analysis showed that this strain could transit from a colistin-sensitive to a resistant phenotype by inserting an IS4 family ISKpn72 element into the colistin-resistance associated mgrB gene. To investigate the mechanism of this transition, we performed genome sequencing analysis of the colistin-sensitive parental strain and found that 12 copies of ISKpn72 containing direct repeats (DR) are located on the chromosome and 1 copy without DR is located on a multidrug-resistant plasmid pR10-341_2. Both types of ISKpn72 could be inserted into the mgrB gene to cause colistin-resistance, though the plasmid-derived ISKpn72 without DR was in higher efficiency. Importantly, we demonstrated that colistin-sensitive K. pneumoniae strain transferred with the ISKpn72 element also obtained the ability to switch from colistin-sensitive to colistin-resistant phenotype. Furthermore, we confirmed that the ISKpn72-containing pR10-341_2 plasmid was able to conjugate, suggesting that the ability of causing colistin-resistant transition is transferable through common conjugation. Our results point to new challenges for both colistin-resistance detection and CRKP treatment.

The objective of this study was to evaluate the effects of wilting and Lactobacillus plantarum (LP) addition on the silage fermentation quality and microbial community of Moringa oleifera Lam. leaf silage. Unwilted (direct-cut) or wilted M. oleifera leaves were prepared either with or without LP (1.0 × 106 cfu/g) followed by either 60 or 120 days of ensiling, leading to eight treatment groups. The results showed that lactic acid was the dominant fermentation product, and no butyric acid was detected for any of the treatments. Higher acetic acid and propionic acid were detected during the fermentation of wilted silage compared to unwilted silage. Although NH3-N content increased after wilting, the content was far below 10% of the dry matter (DM). In addition, higher pH was observed after 120 days of ensiling compared to 60 days. Wilting also influenced the bacterial community structure. Lactobacillus was the most dominant genus in unwilted samples while Enterobacteriales, Weissella, and Pantoea were the most dominant genera in wilted samples. Furthermore, the relative abundance of undesirable microorganisms was far below that of lactic acid bacteria in all treatments. In summary, wilting had significant effects on fermentation quality, and it was shown that M. oleifera leaves can undergo quality ensiling directly without the addition of LP.

This research investigates the impact of long-term nitrogen (N) addition on fluvo-aquic and black soils in north China, with a focus on soil microbial communities and enzyme activities. In each site, there were three N fertilization treatments, i.e., control, moderate-N, and high-N. Phospholipid Fatty Acid Analysis was employed to analyze the microbial community composition, and enzyme activities related to N, carbon (C), and phosphorus (P) cycling were assessed. The results showed that increasing N fertilization levels led to higher soil organic carbon (SOC) and total N (TN) concentrations, indicating enhanced nutrient availability. N fertilization reduced soil pH across both soils, with a more pronounced acidification effect observed in the black soil. Across both soils, N addition increased maize yield, but the higher crop yield was attained in moderate-N rate compared with high-N rate. Microbial community composition analysis revealed that N fertilization induced shifts in the relative abundances of specific microbial groups. The black soil exhibited pronounced shifts in the microbial groups compared to the fluvo-aquic soil, i.e., decreased fungal abundance and fungi: bacteria ratio in response to N input. In addition, the application of N fertilizer led to an elevated ratio of gram-positive to gram-negative (GP:GN) bacteria, but this effect was observed only in black soil. N fertilization had an impact on the enzyme activities related to C, N, and P cycling in both soil types, but black soil showed more pronounced changes in enzyme activities. Permutational multivariate analysis of variance indicated that soil types rather than N fertilization mediated the response of the soil microbial community and enzyme activities. Partial least square path modeling demonstrated that soil pH was the only key driver impacting soil microbial groups and enzyme activities in both soils. In conclusion, our findings highlighted that N fertilization exerted more pronounced impacts on soil biochemical properties, microbial community composition, and enzyme activities in black soil furthermore, moderate N rate resulted in higher crop productivity over high N rate.

The H9N2 virus has been demonstrated to donate its genes to other subtypes of influenza A virus, forming new reassortant virus which may infect human beings. Understanding the genetic characteristic and the global transmission patterns of the virus would guide the prevention and control of potentially emerging avian influenza A virus. In this paper, we hierarchically classified the evolution of the H9N2 virus into three main lineages based on the phylogenetic characteristics of the virus. Due to the distribution of sampling locations, we named the three lineages as Worldwide lineage, Asia-Africa lineage, and China lineage. Codon usage analysis and selective positive site analysis of the lineages further showed the lineage-specific evolution of the virus. We reconstructed the transmission routes of the virus in the three lineages through phylogeography analysis, by which several epicenters for migration of the virus were identified. The hierarchical classification of the lineages implied a possible original seeding process of the virus, starting from the Worldwide lineages to the Asian-Africa lineages and to the China lineages. In the process of H9N2 virus global transmission, the United States was the origin of the virus. China Mainland, Hong Kong SAR, Japan, and Korea were important transfer centers. Based on both the transmission route and the distribution of the hosts in each lineage, we concluded that the wild birds' migration has contributed much to the long-distance global spread of the virus, while poultry trade and people's lifestyle may have contributed to the relatively short-distance transmission in some areas of the Asia and Africa.

In order to screening new Lactic acid bacteria (LAB) strains to alleviating liver injury induced by oxidized oil, we isolated and screened LAB from Chinese fermented foods. Lactobacillus plantarum AR113, Pediococcus pentosaceus AR243, and Lactobacillus plantarum AR501 showed higher scavenging activity of α, α-Diphenyl-β-Picrylhydrazyl (DPPH) free radical and hydrogen radical, stronger inhibition of lipid peroxidation, and better protective effect on yeast cells in vitro. In vivo, oral administration of L. plantarum AR501 improved the antioxidant status of injury mice induced by oxidized oil including decreasing lipid peroxidation, recovering activities of antioxidant enzymes. Meanwhile, the gene expression of Nuclear factor erythroid 2-related factor 2 (Nrf2) of L. plantarum AR501 group was markedly elevated, and several antioxidant genes such as glutathione S-transferase (GSTO1), heme oxygenase-1 (HO-1), Glutamate cysteine ligase (GCL), and NAD(P)H:quinone oxidoreductase-l (NQO1) were subsequently up regulated in mice liver. Therefore, L. plantarum AR501 could be considered as potential candidates for production of functional foods that can alleviate the oxidative damage induced by oxidized oil.

Tuberculosis (TB) is a debilitating infectious disease responsible for more than one million deaths per year. The emergence of drug-resistant TB poses an urgent need for the development of new anti-TB drugs. In this study, we screened a library of over 4,000 small molecules and found that orbifloxacin and the peptide AK15 possess significant bactericidal activity against Mycobacterium tuberculosis (Mtb) in vitro. Orbifloxacin also showed an effective ability on the clearance of intracellular Mtb and protect mice from a strong inflammatory response but not AK15. Moreover, we identified 17 nucleotide mutations responsible for orbifloxacin resistance by whole-genome sequencing. A critical point mutation (D94G) of the DNA gyrase (gyrA) gene was found to be the key role of resistance to orbifloxacin. The computational docking revealed that GyrA D94G point mutation can disrupt the orbifloxacin-protein gyrase interactions mediated by magnesium ion bridge. Overall, this study indicated the potential ability of orbifloxacin as an anti-tuberculosis drug, which can be used either alone or in combination with first-line antibiotics to achieve more effective therapy on TB.

Streptococcus mutans is considered the most relevant bacteria in the transition of non-pathogenic commensal oral microbiota to biofilms which contribute to the dental caries process. The present study aimed to evaluate the antimicrobial activity of a natural plant product, cinnamaldehyde against S. mutans biofilms. Minimum inhibitory concentrations (MIC), minimal bactericidal concentration (MBC), and growth curves were determined to assess its antimicrobial effect against planktonic S. mutans. The biofilm biomass and metabolism with different concentrations of cinnamaldehyde and different incubation time points were assessed using the crystal violet and MTT assays. The biofilms were visualized using confocal laser scanning microscopy (CLSM). Bacterial cell surface hydrophobicity, aggregation, acid production, and acid tolerance were evaluated after cinnamaldehyde treatment. The gene expression of virulence-related factors (gtfB, gtfC, gtfD, gbpB, comDE, vicR, ciaH, ldh and relA) was investigated by real-time PCR. The MIC and MBC of cinnamaldehyde against planktonic S. mutans were 1000 and 2000 μg/mL, respectively. The results showed that cinnamaldehyde can decrease biofilm biomass and metabolism at sub-MIC concentrations. CLSM images revealed that the biofilm-covered surface areas decreased with increasing concentrations of cinnamaldehyde. Cinnamaldehyde increased cell surface hydrophobicity, reduced S. mutans aggregation, inhibited acid production, and acid tolerance. Genes expressions in the biofilms were down-regulated in the presence of cinnamaldehyde. Therefore, our data demonstrated that cinnamaldehyde at sub-MIC level suppressed the microbial activity on S. mutans biofilm by modulating hydrophobicity, aggregation, acid production, acid tolerance, and virulence gene expression.

Tannic acid (TA), a type of polyphenol, is widely distributed in plants, especially in legumes. Not only does it possess antimicrobial properties, but it also has the ability to bind with proteins. The fermentation parameters, nitrogen fractions, antioxidant capacity, and bacterial communities present in mulberry leaves and stylo (Stylosanthes guianensis) ensiled with or without 1 and 2% TA per kilogram of fresh matter (FM) were investigated after 75 days' fermentation. The results showed that 1 and 2% TA both significantly decreased the butyric acid content (4.39 and 7.83 g/kg dry matter (DM), respectively) to an undetectable level in both mulberry leaf and stylo silage. In addition, 2% TA significantly increased the contents of lactate (24.0-39.0 and 8.50-32.3 g/kg DM), acetate (18.0-74.5 and 9.07-53.3 g/kg DM), and the antioxidant capacity of both mulberry leaf and stylo silage, respectively. With the addition of 1 and 2% TA, the pH values (5.55-5.04 and 4.87, respectively) and ammonia-N (NH3-N) content (85.5-27.5 and 16.9 g/kg total nitrogen (TN), respectively) were all significantly decreased in stylo silage. In addition, TA increased the relative abundance of Weissella, Acinetobacter, and Kosakonia spp. and decreased that of undesirable Clostridium spp. TA can thus be used to improve the silage quality of both mulberry leaf and stylo silage, with 2% TA being the better concentration of additive to use.