H7N9 subtype avian influenza viruses (AIVs) cause 1567 human infections and have high mortality, posing a significant threat to public health. Previously, we reported that two avian-derived H7N9 isolates (A/chicken/Eastern China/JTC4/2013 and A/chicken/Eastern China/JTC11/2013) exhibit different pathogenicities in mice. To understand the genetic basis for the differences in virulence, we constructed a series of mutant viruses based on reverse genetics. We found that the PB2-E627K mutation alone was not sufficient to increase the virulence of H7N9 in mice, despite its ability to enhance polymerase activity in mammalian cells. However, combinations with PB1-V719M and/or PA-N444D mutations significantly enhanced H7N9 virulence. Additionally, these combined mutations augmented polymerase activity, thereby intensifying virus replication, inflammatory cytokine expression, and lung injury, ultimately increasing pathogenicity in mice. Overall, this study revealed that virulence in H7N9 is a polygenic trait and identified novel virulence-related residues (PB2-627K combined with PB1-719M and/or PA-444D) in viral ribonucleoprotein (vRNP) complexes. These findings provide new insights into the molecular mechanisms underlying AIV pathogenesis in mammals, with implications for pandemic preparedness and intervention strategies.

Listeriosis is a severe disease caused by the foodborne pathogen Listeria monocytogenes, posing a significant risk to vulnerable populations such as the elderly, pregnant women, and newborns. While relatively uncommon, it has a high global mortality rate of 20-30%. Recent research indicates that smaller outbreaks of the more severe, invasive form of the disease occur more frequently than previously thought, despite the overall stable infection rates of L. monocytogenes over the past 10 years. The ability of L. monocytogenes to form biofilm structures on various surfaces in food production environments contributes to its persistence and challenges in eradication, potentially leading to contamination of food and food production facilities. To address these concerns, this review focuses on recent developments in epidemiology, risk evaluations, and molecular mechanisms of L. monocytogenes survival in adverse conditions and environmental adaptation. Additionally, it covers new insights into strain variability, pathogenicity, mutations, and host vulnerability, emphasizing the important events framework that elucidates the biochemical pathways from ingestion to infection. Understanding the adaptation approaches of L. monocytogenes to environmental stress factors is crucial for the development of effective and affordable pathogen control techniques in the food industry, ensuring the safety of food production.

Cell cycle progression, autophagic cell death during appressorium development, and ROS degradation at the infection site are important for the development of rice blast disease. However, the association of cell cycle, autophagy and ROS detoxification remains largely unknown in M. oryzae. Here, we identify the dual-specificity kinase MoLKH1, which serves as an important cell cycle regulator required for appressorium formation by regulating cytokinesis and cytoskeleton in M. oryzae. MoLKH1 is transcriptionally activated by H2O2 and required for H2O2-induced autophagic cell death and suppression of ROS-activated plant defense during plant invasion of M. oryzae. In addition, the Molkh1 mutant also showed several phenotypic defects, including delayed growth, abnormal conidiation, damaged cell wall integrity, impaired glycogen and lipid transport, reduced secretion of extracellular enzymes and effectors, and attenuated virulence of M. oryzae. Nuclear localization of MoLKH1 requires the nuclear localization sequence, Lammer motif, as well as the kinase active site and ATP-binding site in this protein. Site-directed mutagenesis showed that each of them plays crucial roles in fungal growth and pathogenicity of M. oryzae. In conclusion, our results demonstrate that MoLKH1-mediated cell cycle, autophagy, and suppression of plant immunity play crucial roles in development and pathogenicity of M. oryzae.

Glaesserella parasuis is an important porcine pathogen that commonly colonizes the upper respiratory tract of pigs and is prone to causing Glässer\'s disease under complex conditions. As yet, the disease has led to serious economic losses to the swine industry worldwide. Studies so far have found that several virulence factors are associated with the pathogenicity of G. parasuis, but the pathogenic mechanism is still not fully understood. Cytolethal distending toxin (CDT), a potential virulence factor in G. parasuis, is involved in cytotoxicity, serum resistance, adherence to and invasion of host cells in vitro. Here, to further investigate the pathogenic role of CDT during G. parasuis infection in vitro and in vivo, a double cdt1 and cdt2 deletion mutant (Δcdt1Δcdt2) without selectable marker was first generated in G. parasuis JS0135 strain by continuous natural transformations and replica plating. Morphological observation and lactate dehydrogenase assay showed that the Δcdt1Δcdt2 mutant was defective in cytotoxicity. Additionally, the Δcdt1Δcdt2 mutant was more susceptible to phagocytosis caused by 3D4/2 macrophages compared to the wild-type JS0135 strain. Moreover, by focusing on clinical signs, necropsy, bacterial recovery and pathological observation, we found that the deletion of cdt1 and cdt2 genes led to a significant attenuation of virulence in G. parasuis. Taken together, these findings suggest that as an important virulence factor, CDT can significantly affect the pathogenicity of G. parasuis.

The bacterial species Salmonella enterica (S. enterica) is a highly diverse pathogen containing more than 2600 distinct serovars, which can infect a wide range of animal and human hosts. Recent global emergence of multidrug resistant strains, from serovars Infantis and Muenchen is associated with acquisition of the epidemic megaplasmid, pESI that augments antimicrobial resistance and pathogenicity. One of the main pESI\'s virulence factors is the potent iron uptake system, yersiniabactin encoded by fyuA, irp2-irp1-ybtUTE, ybtA, and ybtPQXS gene cluster. Here we show that yersiniabactin, has an underappreciated distribution among different S. enterica serovars and subspecies, integrated in their chromosome or carried by different conjugative plasmids, including pESI. While the genetic organization and the coding sequence of the yersiniabactin genes are generally conserved, a 201-bp insertion sequence upstream to ybtA, was identified in pESI. Despite this insertion, pESI-encoded yersiniabactin is regulated by YbtA and the ancestral Ferric Uptake Regulator (Fur), which binds directly to the ybtA and irp2 promoters. Furthermore, we show that yersiniabactin genes are specifically induced during the mid-late logarithmic growth phase and in response to iron-starvation or hydrogen peroxide. Concurring, yersiniabactin was found to play a previously unknown role in oxidative stress tolerance and to enhance intestinal colonization of S. Infantis in mice. These results indicate that yersiniabactin contributes to Salmonella fitness and pathogenicity in vivo and is likely to play a role in the rapid dissemination of pESI among globally emerging Salmonella lineages.

In middle to late 2023, a sublineage of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron XBB, EG.5.1 (a progeny of XBB.1.9.2), is spreading rapidly around the world. We performed multiscale investigations, including phylogenetic analysis, epidemic dynamics modeling, infection experiments using pseudoviruses, clinical isolates, and recombinant viruses in cell cultures and experimental animals, and the use of human sera and antiviral compounds, to reveal the virological features of the newly emerging EG.5.1 variant. Our phylogenetic analysis and epidemic dynamics modeling suggested that two hallmark substitutions of EG.5.1, S:F456L and ORF9b:I5T are critical to its increased viral fitness. Experimental investigations on the growth kinetics, sensitivity to clinically available antivirals, fusogenicity, and pathogenicity of EG.5.1 suggested that the virological features of EG.5.1 are comparable to those of XBB.1.5. However, cryo-electron microscopy revealed structural differences between the spike proteins of EG.5.1 and XBB.1.5. We further assessed the impact of ORF9b:I5T on viral features, but it was almost negligible in our experimental setup. Our multiscale investigations provide knowledge for understanding the evolutionary traits of newly emerging pathogenic viruses, including EG.5.1, in the human population.

Avian influenza virus (AIV) infection and vaccination against live attenuated infectious bronchitis virus (aIBV) are frequent in poultry worldwide. Here, we evaluated the clinical effect of H9N2 subtype AIV and QX genotype aIBV co-infection in specific-pathogen-free (SPF) white leghorn chickens and explored the potential mechanisms underlying the observed effects using by 4D-FastDIA-based proteomics. The results showed that co-infection of H9N2 AIV and QX aIBV increased mortality and suppressed the growth of SPF chickens. In particular, severe lesions in the kidneys and slight respiratory signs similar to the symptoms of virulent QX IBV infection were observed in some co-infected chickens, with no such clinical signs observed in single-infected chickens. The replication of H9N2 AIV was significantly enhanced in both the trachea and kidneys, whereas there was only a slight effect on the replication of the QX aIBV. Proteomics analysis showed that the IL-17 signaling pathway was one of the unique pathways enriched in co-infected chickens compared to single infected-chickens. A series of metabolism and immune response-related pathways linked with co-infection were also significantly enriched. Moreover, co-infection of the two pathogens resulted in the enrichment of the negative regulation of telomerase activity. Collectively, our study supports the synergistic effect of the two pathogens, and pointed out that aIBV vaccines might increased IBV-associated lesions due to pathogenic co-infections. Exacerbation of the pathogenicity and mortality in H9N2 AIV and QX aIBV co-infected chickens possibly occurred because of an increase in H9N2 AIV replication, the regulation of telomerase activity, and the disturbance of cell metabolism and the immune system.

Sweet cherry (Prunus avium L.) has become an economically important fruit in China. And its cultivation area has significantly expanded over the last three decades (Wang et al. 2020; Zhao et al. 2023). In July 2023, wilting of cherry trees was observed in a cherry plantation in Wenchuan County (31°51\'N, 103°56\'E, altitude: 1,510 m) in Sichuan Province and approximately 27% of the trees showed symptoms of root rot including soft roots, dark brown to black lesions, yellowing and wilted leaves, and a distinct yellow-brown core discoloration of the inner root core when cut in cross-section. To isolate the causal pathogens, six infected sweet cherry plants with rootstock \'Daqingye\' from Cerasus pseudocerasus were randomly selected from the orchard and then the intertwined diseased and healthy roots (5mm× 5mm × 2mm) were washed with sterile water to remove surface soil. The root samples were surface sterilized with 75% ethanol for 30 seconds and NaClO for 30 seconds and washed three times with distilled water. The disinfected tissues were placed on potato dextrose agar (PDA) and incubated at 27°C in darkness for 5 days (Zhao et al. 2024). A total of nine fungal isolates with similar morphological characteristics were obtained. The colony obtained through single-spore purification displays a red reverse side and a concentric ring pattern on the front, with a sparse surface. Macroconidia were relatively slender with a curve, like sickle shape, 0 to 3 septate measuring (25.8 to 46.1) μm× (4.2 to 7.5) μm, respectively (n=20). The morphological characteristics were consistent with the description of Fusarium spp. (Li et al. 2021). Among these isolates, only HB5 was selected for additional molecular identification. Three target genes, including the internal transcribed spacer (ITS), partial translation elongation factor 1-alpha (TEF), and RNA polymerase second largest subunit (RPB2) were amplified using the primers ITS1/ITS4, TEF1-728/FTEF1-re, and fRPB2-5F/fRPB2-7r, respectively (Groenewald et al. 2013; Carbone and Kohn 1999; Reeb et al. 2004). Sequences of HB5 was deposited in GenBank (ITS, PP388208; TEF, PP580036; RPB2, PP580035). A BLAST search revealed high similarity to those of F. solani sequences with 99%, 100% and 100% respectively (MN013858.1, JF740846.1, OR371902.1), and a multilocus phylogenetic tree was generated to represent the molecular identification results. Pathogenicity studies were conducted on the rootstocks from \'Daqingye\' of Cerasus pseudocerasus in 1 liter plastic flowerpots. The seedlings were incubated in a constant temperature incubator at 25°C with a humidity level of 65% for two weeks. Following the growth of green leaves, 200ml (1x106 spores/ml) of spore suspensions were poured into pots. After 4 weeks of inoculation, the same symptoms of the inoculated plants were observed consistent with those shown in the field , while control plants were inoculated with distill water with asymptomatic. The inoculated pathogen was confirmed both morphologically and molecularly as described earlier, thereby fulfilling Koch\'s postulates. It has been reported that Fusarium solani has been reported to cause root rot in various plants in China, including Actinidia sppt, Zanthoxylum bungeanum, Fragaria×ananassa Duch (Song et al.2022; Li et al. 2023; Zhao et al. 2024). To our knowledge, this is the first report of Fusarium solani causing root rot in sweet cherry (Prunus avium). We here also report the severity and outbreak of this disease, which has been found in other regions in recent years and may become prevalent. Further research on disease management strategies is urgently needed to protect sweet cherry production.

Classical swine fever (CSF) re-emerged in Japan in 2018 for the first time in 26 years. The disease has been known to be caused by a moderately pathogenic virus, rather than the highly pathogenic virus that had occurred in the past. However, the underlying pathophysiology remains unknown. This study conducted an experimental challenge on specific pathogen-free (SPF) pigs in a naïve state for 2, 4, and 6 weeks and confirmed the disease state during each period by clinical observation, virus detection, and pathological necropsy. We revealed the pathological changes and distribution of pathogens and virus-specific antibodies at each period after virus challenge. These results were comprehensively analyzed and approximately 70% of the pigs recovered, especially at 4- and 6-week post-virus challenge. This study provides useful information for future countermeasures against CSF by clarifying the pathogenicity outcomes in unvaccinated pigs with moderately pathogenic genotype 2.1 virus.

Antibiotic resistance, a significant public health hazard, is predicted to cause 10 million deaths worldwide by 2050. The study aimed to identify culturable bioaerosols in the indoor air of dental units in Lahore and assess their antibiotic resistance. Air samples were collected from 10 dental unit locations at different distances, with average concentrations of fungi and bacteria falling within intermediate ranges, per the Global Index of Microbial Contamination (GIMC/m3) index. The study found higher antibiotic-resistant strains in hospital dental units, particularly during winter. The most vigorous strain, S.aureus-NAJIH18, exhibited 70% resistance to ceftazidime. The research highlights the importance of quantifying microbial pollutants for evaluating their source and complexity. It suggests proactive mitigation techniques, such as focused cleaning and air filtration, to improve indoor air quality can mitigate the spread of antibiotic-resistant strains. These insights offer hope in combating the growing public health threat of antibiotic resistance.