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The intracellular pathogen Salmonella enterica serovar Typhimurium (S. Typhimurium) exploits host macrophage as a crucial survival and replicative niche. To minimize host immune response stimulated by flagellin, the expression of flagellar genes is downregulated during S. Typhimurium growth within host macrophages. However, the underlying mechanisms are largely unknown. In this study, we show that STM14_1285 (named AsiR), a putative RpiR-family transcriptional regulator, which is downregulated within macrophages as previously reported and also confirmed here, positively regulates the expression of flagellar genes by directly binding to the promoter of flhDC. By generating an asiR mutant strain and a strain that persistently expresses asiR gene within macrophages, we confirmed that the downregulation of asiR contributes positively to S. Typhimurium replication in macrophages and systemic infection in mice, which could be attributed to decreased flagellar gene expression and therefore reduced flagellin-stimulated secretion of pro-inflammatory cytokines IL-1β and TNF-α. Furthermore, the acidic pH in macrophages is identified as a signal for the downregulation of asiR and therefore flagellar genes. Collectively, our results reveal a novel acidic pH signal-mediated regulatory pathway that is utilized by S. Typhimurium to promote intracellular replication and systemic pathogenesis by repressing flagellar gene expression.

Shigella is an intracellular pathogen that primarily infects the human colon and causes shigellosis. Shigella virulence relies largely on the type III secretion system (T3SS) and secreted effectors. VirF, the master Shigella virulence regulator, is essential for the expression of T3SS-related genes. In this study, we found that YhjC, a LysR-type transcriptional regulator, is required for Shigella virulence through activating the transcription of virF. Pathogenicity of the yhjC mutant, including colonization in the colons of guinea pigs as well as its ability for host cell adhesion and invasion, was significantly lowered. Expression levels of virF and nearly all VirF-dependent genes were downregulated by yhjC deletion, indicating that YhjC can activate virF transcription. Electrophoretic mobility shift assay analysis demonstrated that YhjC could bind directly to the virF promoter region. Therefore, YhjC is a novel virulence regulator that positively regulates the virF expression and promotes Shigella virulence. Additionally, genome-wide expression analysis identified the presence of other genes in the large virulence plasmid and a genome exhibiting differential expression in response to yhjC deletion, with 169 downregulated and 99 upregulated genes, indicating that YhjC also functioned as a global regulatory factor.

Salmonella enterica serovar Typhi (S. Typhi) is a human-limited intracellular pathogen and the cause of typhoid fever, a severe systemic disease. Pathogen–host interaction at the metabolic level affects the pathogenicity of intracellular pathogens, but it remains unclear how S. Typhi infection influences host metabolism for its own benefit. Herein, using metabolomics and transcriptomics analyses, combined with in vitro and in vivo infection assays, we investigated metabolic responses in human macrophages during S. Typhi infection, and the impact of these responses on S. Typhi intracellular replication and systemic pathogenicity. We observed increased glucose content, higher rates of glucose uptake and glycolysis, and decreased oxidative phosphorylation in S. Typhi-infected human primary macrophages. Replication in human macrophages and the bacterial burden in systemic organs of humanized mice were reduced by either the inhibition of host glucose uptake or a mutation of the bacterial glucose uptake system, indicating that S. Typhi utilizes host-derived glucose to enhance intracellular replication and virulence. Thus, S. Typhi promotes its pathogenicity by inducing metabolic changes in host macrophages and utilizing the glucose that subsequently accumulates as a nutrient for intracellular replication. Our findings provide the first metabolic signature of S. Typhi-infected host cells and identifies a new strategy utilized by S. Typhi for intracellular replication.

This study investigated methane production, long-chain fatty acids (LCFAs) profile, and predominant microorganisms in anaerobic digestion (AD) of lipid-rich swine slaughterhouse waste (SSW). The maximum methane yield was 999.2 mL/g VS. LCFAs, as inhibitory hydrolysis products, accumulated first to 1165 mg/L on day 3, and then decreased sharply to 125.7 mg/L on day 9, and finally were degraded to 20 mg/L on day 27. Linoleic acid (C18:2), oleic acid (C18:1) and palmitic acid (C16:0) were the dominant LCFAs. The easy conversion of C18:1 to C16:0 compared with difficult degradation of C16:0 resulted in an increase of C16:0 on day 4–6. Predominant microorganisms were Clostridium, Syntrophomonas and Methanospirillum. This study proved the high methane potential of lipid-rich SSW and gained insights into the degradation process by analysis of intermediates of LCFAs and predominant microorganisms. The results can provide valuable guidance for efficient utilization of this waste to produce methane in future.

A large amount of sheep dung (SD) produced on farms in China is a potential feedstock for production of clean energy in the form of biogas. Similarly, waste paper can be a notable energy source that is worth exploiting. In this study, we assessed the feasibility of co-digesting nitrogen-rich SD with the carbon-rich corrugated board (CB) or waste office paper (OP) in varying volatile solids (VS) ratios, to produce methane. Synergistic effect of co-digesting SD with CB and SD with OP on methane production was found in this study. The highest methane yields of 151.62 and 198.85 mL/g-VS were obtained during the co-digestions of SD with CB at 4:1 ratio (SDCB) and SD with OP at 2:3 ratio (SDOP), respectively. High-throughput 16 S rRNA gene sequencing demonstrated that the microbial diversity and richness in SDCB and SDOP co-digests were higher than in SD and OP mono-digests, respectively. Characteristic bacteria and archaea in the digests were strongly substrate-related and might contribute to methane production. The validated results indicated that methane production through anaerobic co-digestion of SD and waste paper can be an efficient way that could not only reduce environmental pollution but also contribute to methane production.

This study employs a microaerobic pretreatment that includes different microbial reagents combined with oxygen to enhance the energy conversion potential of rice straw (RS) via anaerobic digestion (AD) in a more cost-effective and time-efficient manner. Five broad varieties of microbial reagents including liquid consortium (LC), cow manure (CM), sheep dung (SD), biogas slurry (BS), and straw-decomposing consortia (SC), have been used as sources of useful microbes at varying oxygen concentrations of 0, 6, 12 and 24 mLO2/gVS to explore the effect of microaerobic pretreatment on RS degradation. The results showed that RS pretreated with SC had the highest cumulative methane yield (CMY) of 311.7 mL/gVS at 12 mLO2/gVS, followed by BS, CM, LC, and SD, with the CMYs of 270.5, 263.2, 257.1, and 256.7 mL/gVS at 6 mLO2/gVS, respectively. The RS pretreated with SC, BS, CM, LC, and SD led to improvements in the methane yield by 88.7%, 63.7%, 59.3%, 55.6%, and 55.4%, respectively, compared to the untreated RS. The microbial community analysis revealed that the relative abundance (RA) of Firmicutes, which are known to play a key role in accelerating the rate of hydrolysis, increased after microbial pretreatment combined with oxygen supply. The RA of Clostridium III, which facilitates cell wall degradation via hydrolytic pathways along with other sugar-fermenting bacteria, such as Saccharofermentans, Lutaonella, and Sedimentibacter, increased after AD in the digestate of pretreated RS compared to that in untreated RS. Moreover, a shift in acetoclastic and hydrogenotrophic archaea, including Methanothrix and Methanospirillum, was also observed, indicating precursor changes resulted by pretreatments and varied microbial metabolism, which might have enhanced the methane production process of RS. The energy potential analysis revealed that microbial pretreatment was a suitable technique for stimulating the energy conversion efficiency of RS. This study can be used as a reference for future utilization of a diverse range of lignocellulosic materials for clean energy production.

Tobacco stalk, a common agricultural waste derived from the harvest of tobacco, caused serious environmental pollution in China. In this study, the performance of biomethane production and characteristics of four varieties of tobacco stalk were investigated for the first time. The results showed that the highest cumulative methane yield of 130.2 mL/g-VS was obtained from Nicotiana tabacum L., Yunyan114, which had lower lignin content than other varieties of tobacco stalk. Moreover, different kinetic models were used to describe the biomethane production process, and it was found that the modified Gompertz model was more suitable to simulate the anaerobic digestion (AD) of tobacco stalk. The findings of this study not only showed a feasible method for minimizing the pollution issues of tobacco stalk waste but also gave fundamental information for future AD application.