Strategies adopted by Salmonella to survive in host: a review.

Salmonella, a Gram-negative bacterium that infects humans and animals, causes diseases ranging from gastroenteritis to severe systemic infections. Here, we discuss various strategies used by Salmonella against host cell defenses. Epithelial cell invasion largely depends on a Salmonella pathogenicity island (SPI)-1-encoded type 3 secretion system, a molecular syringe for injecting effector proteins directly into host cells. The internalization of Salmonella into macrophages is primarily driven by phagocytosis. After entering the host cell cytoplasm, Salmonella releases many effectors to achieve intracellular survival and replication using several secretion systems, primarily an SPI-2-encoded type 3 secretion system. Salmonella-containing vacuoles protect Salmonella from contacting bactericidal substances in epithelial cells and macrophages. Salmonella modulates the immunity, metabolism, cell cycle, and viability of host cells to expand its survival in the host, and the intracellular environment of Salmonella-infected cells promotes its virulence. This review provides insights into how Salmonella subverts host cell defenses for survival.

The LysR-Type Transcription Regulator YhjC Promotes the Systemic Infection of Salmonella Typhimurium in Mice.

Salmonella Typhimurium is a Gram-negative intestinal pathogen that can infect humans and a variety of animals, causing gastroenteritis or serious systemic infection. Replication within host macrophages is essential for S. Typhimurium to cause systemic infection. By analyzing transcriptome data, the expression of yhjC gene, which encodes a putative regulator in S. Typhimurium, was found to be significantly up-regulated after the internalization of Salmonella by macrophages. Whether yhjC gene is involved in S. Typhimurium systemic infection and the related mechanisms were investigated in this study. The deletion of yhjC reduced the replication ability of S. Typhimurium in macrophages and decreased the colonization of S. Typhimurium in mouse systemic organs (liver and spleen), while increasing the survival rate of the infected mice, suggesting that YhjC protein promotes systemic infection by S. Typhimurium. Furthermore, by using transcriptome sequencing and RT-qPCR assay, the transcription of several virulence genes, including spvD, iroCDE and zraP, was found to be down-regulated after the deletion of yhjC. Electrophoretic mobility shift assay showed that YhjC protein can directly bind to the promoter region of spvD and zraP to promote their transcription. These findings suggest that YhjC contributes to the systemic virulence of S. Typhimurium via the regulation of multiple virulence genes and YhjC could represent a promising target to control S. Typhimurium infection.

Nitrate Utilization Promotes Systemic Infection of Salmonella Typhimurium in Mice.

Salmonella Typhimurium is an invasive enteric pathogen that causes gastroenteritis in humans and life-threatening systemic infections in mice. During infection of the intestine, S. Typhimurium can exploit nitrate as an electron acceptor to enhance its growth. However, the roles of nitrate on S. Typhimurium systemic infection are unknown. In this study, nitrate levels were found to be significantly increased in the liver and spleen of mice systemically infected by S. Typhimurium. Mutations in genes encoding nitrate transmembrane transporter (narK) or nitrate-producing flavohemoprotein (hmpA) decreased the replication of S. Typhimurium in macrophages and reduced systemic infection in vivo, suggesting that nitrate utilization promotes S. Typhimurium systemic virulence. Moreover, nitrate utilization contributes to the acidification of the S. Typhimurium cytoplasm, which can sustain the virulence of S. Typhimurium by increasing the transcription of virulence genes encoding on Salmonella pathogenicity island 2 (SPI-2). Furthermore, the growth advantage of S. Typhimurium conferred by nitrate utilization occurred only under low-oxygen conditions, and the nitrate utilization was activated by both the global regulator Fnr and the nitrate-sensing two-component system NarX-NarL. Collectively, this study revealed a novel mechanism adopted by Salmonella to interact with its host and increase its virulence.

R-CenterNet+: Anchor-Free Detector for Ship Detection in SAR Images.

In recent years, the rapid development of Deep Learning (DL) has provided a new method for ship detection in Synthetic Aperture Radar (SAR) images. However, there are still four challenges in this task. (1) The ship targets in SAR images are very sparse. A large number of unnecessary anchor boxes may be generated on the feature map when using traditional anchor-based detection models, which could greatly increase the amount of computation and make it difficult to achieve real-time rapid detection. (2) The size of the ship targets in SAR images is relatively small. Most of the detection methods have poor performance on small ships in large scenes. (3) The terrestrial background in SAR images is very complicated. Ship targets are susceptible to interference from complex backgrounds, and there are serious false detections and missed detections. (4) The ship targets in SAR images are characterized by a large aspect ratio, arbitrary direction and dense arrangement. Traditional horizontal box detection can cause non-target areas to interfere with the extraction of ship features, and it is difficult to accurately express the length, width and axial information of ship targets. To solve these problems, we propose an effective lightweight anchor-free detector called R-Centernet+ in the paper. Its features are as follows: the Convolutional Block Attention Module (CBAM) is introduced to the backbone network to improve the focusing ability on small ships; the Foreground Enhance Module (FEM) is used to introduce foreground information to reduce the interference of the complex background; the detection head that can output the ship angle map is designed to realize the rotation detection of ship targets. To verify the validity of the proposed model in this paper, experiments are performed on two public SAR image datasets, i.e., SAR Ship Detection Dataset (SSDD) and AIR-SARShip. The results show that the proposed R-Centernet+ detector can detect both inshore and offshore ships with higher accuracy than traditional models with an average precision of 95.11% on SSDD and 84.89% on AIR-SARShip, and the detection speed is quite fast with 33 frames per second.

Salmonella enterica Serovar Typhi Induces Host Metabolic Reprogramming to Increase Glucose Availability for Intracellular Replication.

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.

Antifungal efficacy of scorpion derived peptide K1K8 against Candida albicans in vitro and in vivo.

As an alternative class of antimicrobial agents, antimicrobial peptides (AMPs) have gained significant attention. In this study, K1K8, a scorpion AMP derivative, showed effective activity against Candida albicans including clinically resistant strains. K1K8 killed C. albicans cells mainly by damaging the cell membrane and inducing necrosis via an ROS-related pathway. K1K8 could also interact with DNA after damaging the nuclear envelope. Moreover, K1K8 inhibited hyphal development and biofilm formation of C. albicans in a dose-dependent manner. In the mouse skin infection model, K1K8 significantly decreased the counts of C. albicans cells in the infection area. Overall, K1K8 is a potential anti-infective agent against skin infections caused by C. albicans.

CobB-mediated deacetylation of the chaperone CesA regulates Escherichia coli O157:H7 virulence.

Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a common food-borne pathogen that can cause acute diseases. Lysine acetylation is a post-translational modification (PTM) that occurs in various prokaryotes and is regulated by CobB, the only deacetylase found in bacteria. Here, we demonstrated that CobB plays an important role in the virulence of EHEC O157:H7 and that deletion of cobB significantly decreased the intestinal colonization ability of bacteria. Using acetylation proteomic studies, we systematically identified several proteins that could be regulated by CobB in EHEC O157:H7. Among these CobB substrates, we found that acetylation at the K44 site of CesA, a chaperone for the type-III secretion system (T3SS) translocator protein EspA, weakens its binding to EspA, thereby reducing the stability of this virulence factor; this PTM ultimately attenuating the virulence of EHEC O157:H7. Furthermore, we showed that deacetylation of the K44 site, which is deacetylated by CobB, promotes the interaction between CesA and EspA, thereby increasing bacterial virulence in vitro and in animal experiments. In summary, we showed that acetylation influences the virulence of EHEC O157:H7, and uncovered the mechanism by which CobB contributes to bacterial virulence based on the regulation of CesA deacetylation.

Nitric oxide is a host cue for Salmonella Typhimurium systemic infection in mice.

Nitric oxide (NO) is produced as an innate immune response against microbial infections. Salmonella Typhimurium (S. Typhimurium), the major causative pathogen of human gastroenteritis, induces more severe systemic disease in mice. However, host factors contributing to the difference in species-related virulence are unknown. Here, we report that host NO production promotes S. Typhimurium replication in mouse macrophages at the early infection stage by activating Salmonella pathogenicity island-2 (SPI-2). The NO signaling-induced SPI-2 activation is mediated by Fnr and PhoP/Q two-component system. NO significantly induced fnr transcription, while Fnr directly activated phoP/Q transcription. Mouse infection assays revealed a NO-dependent increase in bacterial burden in systemic organs during the initial days of infection, indicating an early contribution of host NO to virulence. This study reveals a host signaling-mediated virulence activation pathway in S. Typhimurium that contributes significantly to its systemic infection in mice, providing further insights into Salmonella pathogenesis and host-pathogen interaction.

Anaerobic and Microaerobic Pretreatment for Improving Methane Production From Paper Waste in Anaerobic Digestion.

Having been generated with a tremendous amount annually, paper waste (PW) represents a large proportion in municipal solid waste (MSW) and also a potential source of renewable energy production through the application of anaerobic digestion (AD). However, the recalcitrant lignocellulosic structure poses obstacles to efficient utilization in this way. Recently, anaerobic and microaerobic pretreatment have attracted attention as approaches to overcome the obstacles of biogas production. This study was set out to present a systematic comparison and assessment of anaerobic and microaerobic pretreatment of PW with different oxygen loadings by five microbial agents: composting inoculum (CI), straw-decomposing inoculum (SI), cow manure (CM), sheep manure (SM), and digestate effluent (DE). The hints of microbial community evolution during the pretreatment and AD were tracked by 16S rRNA high-throughput sequencing. The results demonstrated that PW pretreated by DE with an oxygen loading of 15 ml/gVS showed the highest cumulative methane yield (CMY) of 343.2 ml/gVS, with a BD of 79.3%. In addition to DE, SI and SM were also regarded as outstanding microbial agents for pretreatment because of the acceleration of methane production at the early stage of AD. The microbial community analysis showed that Clostridium sensu stricto 1 and Clostridium sensu stricto 10 possessed high relative abundance after anaerobic pretreatment by SI, while Bacteroides and Macellibacteroides were enriched after microaerobic pretreatment by SM, which were all contributable to the cellulose degradation. Besides, aerobic Bacillus in SI and Acinetobacter in SM and DE probably promoted lignin degradation only under microaerobic conditions. During AD, VadinBC27, Ruminococcaceae Incertae Sedis, Clostridium sensu stricto 1, Fastidiosipila, and Caldicoprobacter were the crucial bacteria that facilitated the biodegradation of PW. By comparing the groups with same microbial agent, it could be found that changing the oxygen loading might result in the alternation between hydrogenotrophic and acetoclastic methanogens, which possibly affected the methanogenesis stage. This study not only devised a promising tactic for making full use of PW but also provided a greater understanding of the evolution of microbial community in the pretreatment and AD processes, targeting the efficient utilization of lignocellulosic biomass in full-scale applications.

Downregulation of a novel flagellar synthesis regulator AsiR promotes intracellular replication and systemic pathogenicity of Salmonella typhimurium.

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-1ß 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.

YhjC is a novel transcriptional regulator required for Shigella flexneri virulence.

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 Typhimurium reprograms macrophage metabolism via T3SS effector SopE2 to promote intracellular replication and virulence.

Salmonella Typhimurium establishes systemic infection by replicating in host macrophages. Here we show that macrophages infected with S. Typhimurium exhibit upregulated glycolysis and decreased serine synthesis, leading to accumulation of glycolytic intermediates. The effects on serine synthesis are mediated by bacterial protein SopE2, a type III secretion system (T3SS) effector encoded in pathogenicity island SPI-1. The changes in host metabolism promote intracellular replication of S. Typhimurium via two mechanisms: decreased glucose levels lead to upregulated bacterial uptake of 2- and 3-phosphoglycerate and phosphoenolpyruvate (carbon sources), while increased pyruvate and lactate levels induce upregulation of another pathogenicity island, SPI-2, known to encode virulence factors. Pharmacological or genetic inhibition of host glycolysis, activation of host serine synthesis, or deletion of either the bacterial transport or signal sensor systems for those host glycolytic intermediates impairs S. Typhimurium replication or virulence.

Anaerobic digestion of tobacco stalk: biomethane production performance and kinetic analysis.

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.

Steam explosion pretreatment of rice straw to improve structural carbohydrates anaerobic digestibility for biomethanation.

Effectiveness of steam explosion (SE) pretreatment for deconstructing the complex structural carbohydrates (SC) and lignin recalcitrance properties of rice straw (RS) for conjunctive improvement of biofuel yield and waste valorization was evaluated. This work exhibited successful pretreatment of RS at a different pressure (1.2, 1.5, and 1.8 MPa) and retention (3, 6, 9, and 12 min) for enhancement of SC contribution to biomethane production. Regression analysis demonstrated that SE pretreatment efficiency improved at high-temperature and short-retention time for biodegradation of RS. Maximum cumulative methane yield (EMY) achieved 254.8 mL/gvs at 1.2 MPa (3 min) of SE-treated RS with 62.7% of very significant improvement compared with untreated RS (156.6 mL/gvs). Furthermore, solid fraction of xylose, arabinose, cellobiose, glucose, and acid-soluble lignin in SE-treated RS of 1.2 MPa (3 min) were biodegraded by 27.4%, 46.4%, 100%, 48.8%, and 14.1%, respectively, after anaerobic digestion. Therefore, SE pretreatment was an encouraging approach for enhancing SC conversion to biomethane and waste resource to circular economy.

Anaerobic digestion of lipid-rich swine slaughterhouse waste: Methane production performance, long-chain fatty acids profile and predominant microorganisms.

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.

Employing response surface methodology (RSM) to improve methane production from cotton stalk.

China is the largest cotton producer with the cotton output accounting for 25% of the total world's cotton production. A large quantity of cotton stalk (CS) waste is generated which is burned and causes environmental and ecological problems. This study investigated the anaerobic digestibility of CS by focusing on improving the methane yield by applying central composite design of response surface methodology (RSM). The purpose of this study was to determine the best level of factors to optimize the desired output of methane production from CS. Thus, it was necessary to describe the relationship of many individual variables with one or more response values for the effective utilization of CS. The influences of feed to inoculum (F/I) ratio and organic loading (OL) on methane production were investigated. Results showed that the experimental methane yield (EMY) and volatile solid (VS) removal were calculated to be 70.22 mL/gVS and 14.33% at F/I ratio of 0.79 and organic loading of 25.61 gVS/L, respectively. Characteristics of final effluent showed that the anaerobic system was stable. This research laid a foundation for future application of CS to alleviate the problems of waste pollution and energy output.

In-situ injection of potassium hydroxide into briquetted wheat straw and meadow grass - Effect on biomethane production.

Alkaline pretreatment of lignocellulosic biomass has been intensively investigated but heavy water usage and environmental pollution from wastewater limits its industrial application. This study presents a pretreatment technique by in-situ injection of potassium hydroxide concentrations ranging from 0.8% to 10% (w/w) into the briquetting process of wheat straw and meadow grass. Results show that the biomethane yield and hydrolysis rate was improved significantly with a higher impact on wheat straw compared to meadow grass. The highest biomethane yield from wheat straw briquettes of 353mL.g-1 VS was obtained with 6.27% (w/w) potassium hydroxide injection, which was 14% higher than from untreated wheat straw. The hydrolysis rates of wheat straw and meadow grass increased from 4.27×10-2 to 5.32×10-2d-1 and 4.19×10-2 to 6.00×10-2d-1, respectively. The low water usage and no wastewater production make this a promising technology.