The impact of vaccines extends beyond the improvement of individual health, significantly lowering the burden of infectious disease in healthcare systems and strengthening the societal infrastructure and economic stability of a nation. Unfortunately, the substantial benefits of vaccines are often disproportionately encountered in affluent nations, with the need for cold-chain logistics heavily contributing to this inequity. Here, we describe the biomimetic mineralization of a live viral vaccine using metal-organic frameworks (MOFs) to alleviate stress caused by ambient temperatures. Utilizing a zinc-based MOF (ZIF-8), we demonstrated the advantages of a higher concentration of the ZIF-8 precursor constituents for coverage of viral antigens and recovery of the live viral Newcastle Disease Virus (NDV) vaccine. Using a candidate MOF-based formulation, we identified that ZIF-8 can maintain effective coverage of NDV V4 virions following lyophilization; however, viral titer was negatively impacted if rapid freezing was integrated. As an alternative cost-effective storage technique, we utilized filter paper drying for long-term stabilization of various ZIF-8@NDV formulations. We demonstrated that the presence of ZIF-8 significantly contributed to vaccine stability when stored at 25 or 37 °C compared to NDV in solution. In summary, our findings highlight the potential of utilizing MOFs for stabilization of live viral vaccines at ambient temperatures and demonstrate the viability of filter paper as an alternative to lyophilization for vaccine preservation.

Anaesthesia is sometimes required for the effective restraint of laboratory pigs for sample collection. Yet, anaesthesia can initiate a range of physiological disruptions that can increase variability in study data and lead to poorer animal welfare. Judicious use of anaesthesia can mitigate experimental, human safety, and animal welfare concerns, but it does not eliminate the potential for adverse effects. The use of reversal agents can shorten recovery time and reduce the physiological impacts of anaesthesia but can also cause additional side effects. We, therefore, trialled the use of low-dose atipamezole (0.12 mg/kg) for the antagonism of xylazine in laboratory pigs anaesthetised using a combination of xylazine and zolazepam/tiletamine. We measured time to recovery, selected clinical variables, recovery characteristics, and behaviours to investigate if a low dose of antagonist decreased recovery time and reduced the physiological impacts of anaesthesia whilst avoiding adverse negative side effects. We categorised side effects and behaviours as having either a low or high negative welfare impact based on the potential risk of injury and whether behaviours were displayed before or after return to consciousness. Collectively, our results indicated that while the use of low-dose atipamezole decreased recovery time and improved thermoregulation in most pigs, it introduced and exacerbated adverse side effects and behaviours that can lead to poorer welfare outcomes for laboratory pigs.

The advancement of genetic engineering in chickens has enabled significant advancement in developmental biology, bioreactors, and disease resilience. The development of CRISPR/Cas9 genome engineering technology has further expanded the potential applications of genetic engineering in poultry. In this study we aimed to evaluate the efficacy of a direct in vivo transfection method, previously demonstrated to produce transgenic chickens, in generating gene knockout (KO) chickens. Specifically, we targeted the Interferon-α/β Receptor 1 (IFNAR1) and Interleukin 1 receptor, type I (IL1R1), both critical pathways in the inflammatory and antiviral responses. We designed guide RNAs targeting the genes and validated their efficiency in vivo via microinjection into the developing embryos. PCR analysis confirmed the presence of gene deletions in chimeric roosters, which were subsequently bred to produce G1 germline heterozygote KO offspring. Homozygous KO chickens were generated and subjected to phenotypic and functional analyses. Our results demonstrated successful generation of functional knockouts of both IFNAR1 and IL1R1 using a direct in vivo transfection. Overall, this study demonstrates that direct in vivo transfection provides a robust and predictable method for generating KO chickens, facilitating further research into avian immune responses and the development of antiviral strategies.

mRNA vaccines have emerged as a highly effective strategy for the prevention and treatment of various diseases. A critical factor driving the success of mRNA vaccines is the development of advanced multicomponent lipid nanoparticles (LNPs) as a delivery system. As mRNA-LNP technology becomes increasingly integral to vaccine and therapeutic development, there is significant potential to enhance LNP efficiency and build upon the first generation of clinically approved mRNA-LNP products. This can lead to the development of superior formulations that achieve higher protein expression and improved therapeutic outcomes. In this study, we present a novel approach to enhance the transfection efficiency of mRNA-LNPs using ZIF-8 metal-organic framework (MOF). We demonstrate effective encapsulation of mRNA-LNPs within ZIF-8, with preserved structural integrity during dissociation and release. Remarkably, following MOF encapsulation and release, we observed a 3-fold and 8-fold increase in transfection efficiency of the mRNA-LNPs at 48 h in HEK-293 and HCT-116 cells, respectively. Our findings suggest that the presence of ZIF-8 materials with the mRNA-LNPs significantly contributes to their improved transfection and translation efficiency.

Positive-reinforcement training of laboratory pigs can reduce the reliance on forced manual restraint and anaesthesia for sample collection, reducing stress and physiological disruption. Training regimens for laboratory pigs typically rely on specialised equipment for restraint, such as Panepinto slings, with a time investment that may not be justified for short-term studies. These training regimens also commonly rely on pigs being lifted into sling restraints, which is not practical for studies involving large pigs. We developed and assessed a rapid, three-phase, positive-reinforcement training regimen for both individually housed and group-housed laboratory pigs to facilitate the collection of minimally invasive samples consciously and voluntarily. The time to complete each phase of training in both individually housed and group-housed pigs was recorded. The behaviour of the individually housed pigs was assessed via an ethogram of behaviours exhibited during a human approach test, and stress response was assessed by analysing salivary corticosterone. The rapid, positive-reinforcement training regimen successfully facilitated oral swabbing, rectal swabbing and rectal thermometer insertion from individually housed (within 18 days) and group-housed (within 6 days) pigs. The trained pigs displayed increasing positive behaviours, no or very few negative behaviours and corticosterone levels within normal limits throughout the study. This training regimen provides a practical and welfare-positive tool for the collection of minimally invasive samples from both small and large laboratory pigs, with a low time investment of 2–5 min/pig/day without the need for specialised restraint equipment.

Effective manual or chemical restraint of pigs for blood collection is crucial when conducting infectious disease research under high biocontainment laboratory conditions; however, these methods can introduce stress and physiological disruption. We aimed to establish proof-of-concept for the use of jugular vein vascular access ports (VAPs) for repeated, conscious blood collection in both individually and group housed pigs, trained using a positive reinforcement training regime. In study 1, pigs were housed individually, with three pigs implanted with VAPs. An additional control pig was not implanted with VAPs and was anaesthetised for all blood collections. Daily assessments of behaviour were conducted and saliva collected for corticosterone analysis. In study 2, pigs were housed in a group, with three pigs implanted with VAPs. Three control pigs were not implanted with VAPs and were anaesthetised for all blood collections. To assess the physiological impacts of both blood collection methods, heart rate variability, pulse rate and rectal temperature were assessed during a blood collection event. Across both studies 31 of 33 VAP blood collections were successful, with the remaining two collections successful using light sedation. An initial time and cost investment was required for VAP implantation, but the time required to collect blood via VAPs was halved compared to anaesthetising pigs for blood collection. One pig died during VAP surgery from anaesthesia-related complications. Pigs implanted with VAPs displayed low negative and high positive behaviour scores, low salivary corticosterone levels, and maintained steady clinical parameters during a blood collection event, whilst anaesthetised pigs displayed reduced HRV and temperature and increased pulse rate. This study provides proof-of-concept and recommendations for using VAPs in trained pigs under high biocontainment, as a welfare-positive method of conscious, repeated blood collection.

Wearable and implantable physiological monitors are designed for minimally invasive health assessment in companion animals. We aimed to establish proof-of-concept for the use of PetPace collar monitors and VetChip implantable monitoring devices in pigs. PetPace monitors were used to measure pulse rate, respiratory rate and heart rate variability in individually housed pigs. Group housed pigs were fitted with PetPace monitors to assess their feasibility for use in the presence of conspecifics, in addition to implantation with VetChip subcutaneous monitoring devices. PetPace monitors caused skin abrasion after 10 days of continual wear, but simple modifications allowed collars to be worn safely in individually housed pigs. Collars could not be worn unsupervised by group housed pigs due to damage by conspecifics. PetPace pulse and respiratory rate readouts showed a strong correlation with manual auscultations, and PetPace heart rate variability showed a very strong negative correlation with salivary corticosterone levels. VetChip temperature and pulse rate readouts were accurate in healthy pigs. However, the suitability of dorsal mid-scapula VetChip implantation is questionable due to wound breakdown in two out of three pigs.

Over the last 20 million years, the Andropogoneae tribe of grasses has evolved to dominate 17% of global land area. Domestication of these grasses in the last 10,000 years has yielded our most productive crops, including maize, sugarcane, and sorghum. The majority of Andropogoneae species, including maize, show a history of polyploidy - a condition that, while offering the evolutionary advantage of multiple gene copies, poses challenges to basic cellular processes, gene expression, and epigenetic regulation. Genomic studies of polyploidy have been limited by sparse sampling of taxa in groups with multiple polyploidy events. Here, we present 33 genome assemblies from 27 species, including chromosome-scale assemblies of maize relatives Zea and Tripsacum. In maize, the after-effects of polyploidy have been widely studied, showing reduced chromosome number, biased fractionation of duplicate genes, and transposable element (TE) expansions. While we observe these patterns within the genus Zea, 12 other polyploidy events deviate significantly. Those tetraploids and hexaploids retain elevated chromosome number, maintain nearly complete complements of duplicate genes, and have only stochastic TE amplifications. These genomes reveal variable outcomes of polyploidy, challenging simple predictions and providing a foundation for understanding its evolutionary implications in an ecologically and economically important clade.

The CRISPR-Cas13 system has emerged as a powerful platform for programmable RNA targeting, offering efficient and sequence-specific silencing of coding and non-coding transcripts. The RNA-targeting capabilities of CRISPR-Cas13 have been harnessed to silence transcripts harbouring pathogenic mutations and combat infectious diseases. However, the molecular basis of on-target and collateral activity are not completely understood, limiting the utility of Cas13 systems. In this study, we delineate the principles for the development of effective crRNAs by targeting DsRed fluorescence reporter and synthetic influenza mRNA in chicken fibroblast DF1 cells. To systematically determine the optimal design for RfxCas13d crRNA, we investigated the minimum length of the crRNA, importance of protospacer flanking sequence, degree of mismatch tolerance, and off target effects. Our data reveal variable knockdown levels between crRNAs, in which several crRNAs achieved over 95% target knockdown. We show that crRNAs exhibit a high degree of tolerance to single-nucleotide mismatches, regardless of their position in the spacer sequence. However, 4-nt mismatches between the spacer and the target significantly reduces targeting efficacy, whereas eight nucleotide mismatches completely abolish the activity of RfxCas13d. Finally, we compared targeting efficiency and collateral activity of two widely used RfxCas13d and HfCas13d variants. Our data extend current understanding of Cas13d-mediated RNA targeting and offer a framework for rational crRNA design to enhance effectiveness in diverse applications, including antiviral strategies.

Enhancing livestock biosecurity is critical to safeguard the livelihoods of farmers, global and local economies, and food security. Vaccination is fundamental to the control and prevention of exotic and endemic high-priority infectious livestock diseases. Successful implementation of vaccination in a biosecurity plan is underpinned by a strong understanding of correlates of protection—those elements of the immune response that can reliably predict the level of protection from viral challenge. While correlates of protection have been successfully characterized for many human viral vaccines, for many high-priority livestock viral diseases, including African swine fever and foot and mouth disease, they remain largely uncharacterized. Current literature provides insights into potential correlates of protection that should be assessed during vaccine development for these high-priority mammalian livestock viral diseases. Establishment of correlates of protection for biosecurity purposes enables immune surveillance, rationale for vaccine development, and successful implementation of livestock vaccines as part of a biosecurity strategy.

Inflammation and lipid regulator with UBA-like and NBR1-like domains (ILRUN) is a protein-encoding gene associated with innate immune signaling, lipid metabolism and cancer. In the context of innate immunity, ILRUN inhibits IRF3-mediated transcription of antimicrobial and proinflammatory cytokines by inducing degradation of the transcriptional coactivators CBP and p300. There remains a paucity of information, however, regarding the innate immune roles of ILRUN beyond in vitro analyses. To address this, we utilize a knockout mouse model to investigate the effect of ILRUN on cytokine expression in splenocytes and on the development of immune cell populations in the spleen and thymus. We show elevated production of tumor necrosis factor and interleukin-6 cytokines in ILRUN-deficient splenocytes following stimulation with the innate immune ligands polyinosinic: polycytidylic acid or lipopolysaccharide. Differences were also observed in the populations of several T cell subsets, including regulatory, mucosal-associated invariant and natural killer. These data identify novel functions for ILRUN in the development of certain immune cell populations and support previous in vitro findings that ILRUN negatively regulates the synthesis of pathogen-stimulated cytokines. This establishes the ILRUN knockout mouse model as a valuable resource for further study of the functions of ILRUN in health and disease.

Stress and general anesthesia have an impact on the functional response of the organism due to the detrimental effects on cardiovascular, immunological, and metabolic function, which could limit the organism's response to an infectious event. Animal studies have formed an essential step in understanding and mitigating infectious diseases, as the complexities of physiology and immunity cannot yet be replicated in vivo. Using animals in research continues to come under increasing societal scrutiny, and it is therefore crucial that the welfare of animals used in disease research is optimized to meet both societal expectations and improve scientific outcomes. Everyday management and procedures in animal studies are known to cause stress, which can not only cause poorer welfare outcomes, but also introduces variables in disease studies. Whilst general anesthesia is necessary at times to reduce stress and enhance animal welfare in disease research, evidence of physiological and immunological disruption caused by general anesthesia is increasing. To better understand and quantify the effects of stress and anesthesia on disease study and welfare outcomes, utilizing the most appropriate animal monitoring strategies is imperative. This article aims to analyze recent scientific evidence about the impact of stress and anesthesia as uncontrolled variables, as well as reviewing monitoring strategies and technologies in animal models during infectious diseases.

Salmonella Weltevreden is a pathogen associated with human diarrheal diseases found across the globe. However, knowledge of the genomics and epidemiology of this pathogen is still limited.

Influenza A viruses (IAV) pose a constant threat to human and poultry health. Of particular interest are the infections caused by highly pathogenic avian influenza (HPAI) viruses, such as H5N1, which cause significant production issues. In response to influenza infection, cells activate immune mechanisms that lead to increased interferon (IFN) production. To investigate how alterations in the interferon signaling pathway affect the cellular response to infection in the chicken, we used CRISPR/Cas9 to generate a chicken cell line that lacks a functional the type I interferon receptor (IFNAR1). We then assessed viral infections with the WSN strain of influenza. Cells lacking a functional IFNAR1 receptor showed reduced expression of the interferon stimulated genes (ISG) such as Protein Kinase R (PKR) and Myxovirus resistance (Mx) and were more susceptible to viral infection with WSN. We further investigated the role or IFNAR1 on low pathogenicity avian influenza (LPAI) strains (H7N9) and a HPAI strain (H5N1). Intriguingly, Ifnar−/− cells appeared more resistant than WT cells when infected with HPAI virus, potentially indicating a different interaction between H5N1 and the IFN signaling pathway. Our findings support that ChIFNAR1 is a key component of the chicken IFN signaling pathway and these data add contributions to the field of host-avian pathogen interaction and innate immunity in chickens.

The zoonotic H7N9 avian influenza (AI) virus first emerged in 2013 as a low pathogenic (LPAI) strain, and has repeatedly caused human infection resulting in severe respiratory illness and a mortality of ~39% (>600 deaths) across five epidemic waves. This virus has circulated in poultry with little to no discernible clinical signs, making detection and control difficult. Contrary to published data, our group has observed a subset of specific pathogen free chickens infected with the H7N9 virus succumb to disease, showing clinical signs consistent with highly pathogenic AI (HPAI). Viral genome sequencing revealed two key mutations had occurred following infection in the haemagglutinin (HA 226 L>Q) and nucleoprotein (NP 373 A>T) proteins. We further investigated the impact of the NP mutation and demonstrated that only chickens bearing a single nucleotide polymorphism (SNP) in their IFITM1 gene were susceptible to the H7N9 virus. Susceptible chickens demonstrated a distinct loss of CD8 + T cells from the periphery as well as a dysregulation of IFNγ that was not observed for resistant chickens, suggesting a role for the NP mutation in altered T cell activation. Alternatively, it is possible that this mutation led to altered polymerase activity, as the mutation occurs in the NP 360-373 loop which has been previously show to be important in RNA binding. These data have broad ramifications for our understanding of the pathobiology of AI in chickens and humans and provide an excellent model for investigating the role of antiviral genes in a natural host species.

The invaluable health, economic and social impacts of vaccination are hard to exaggerate. The ability to stabilize vaccines is urgently required for their equitable distribution without the dependence on the 'cold-chain' logistics. Herein, for the first time we report biomimetic-mineralization of live-viral vaccines using metal-organic frameworks (MOFs) to enhance their storage stability from days to months. Apply-ing ZIF-8 and aluminium fumarate (Alfum), the Newcastle Disease Virus (NDV) V4 strain and Influenza A WSN strain were encapsulated with remarkable retention of their viral titre. The ZIF-8@NDV, ZIF-8@WSN and Alfum@WSN composites were validated for live-virus recovery using a tissue culture infectious dose (TCID50) assay. With the objective of long-term stabilization, we developed a novel, trehalose (T) and skim milk (SM) stabilized, freeze-dried MOF@Vaccine composite, ZIF-8@NDV + T/SM. The thermal stability of this composite was investigated and compared with the control NDV and non-encapsulated, freeze-dried NDV + T/SM composite at 4 ?, RT, and 37 & DEG;C over a period of 12 weeks. We demonstrate the fragility of the control NDV vaccine which lost all viability at RT and 37 ? by 12 and 4 weeks, respectively. Comparing the freeze-dried counterparts, the MOF encapsulated ZIF-8@NDV + T/SM demonstrated significant en-hancement in stability of the NDV + T/SM composite especially at RT and 37 ? upto 12 weeks.Statement of significanceVaccination is undoubtedly one of the most effective medical interventions, saving millions of lives each year. However, the requirement of 'cold-chain' logistics is a major impediment to widespread immunization. Live viral vaccines (LVVs) are widely used vaccine types with proven efficacy and low cost. Nonetheless, their complex composition increases their susceptability to thermal stress. Several LVV ther-mostabilization approaches have been investigated, including their complex engineering and the facile addition of stabilizers. Still, the lack of a universal approach urgently requires finding a stabilization tech-nique especially when additives alone may not be sufficient. Herein, we demonstrate MOF biomimetic-mineralization technology to encapsulate LVVs developing an optimised composite which significantly preserves vaccines without refrigeration for extended periods of time. (C)& nbsp;2022 The Authors. Published by Elsevier Ltd on behalf of Acta Materialia Inc.& nbsp;

IFITM proteins can prevent the entry of several pH-dependent viruses, including high-consequence viruses such as HIV, influenza viruses, and SARS-coronaviruses. Mutations in these genes have been associated with worsened disease outcomes with mutations in their IFITM genes, highlighting these genes as potential disease risk factors. ABSTRACT The current fears of a future influenza pandemic have resulted in an increased emphasis on the development and testing of novel therapeutic strategies against the virus. Fundamental to this is the ferret model of influenza infection, which is critical in examining pathogenesis and treatment. Nevertheless, a precise evaluation of the efficacy of any treatment strategy in ferrets is reliant on understanding the immune response in this model. Interferon-inducible transmembrane proteins (IFITMs) are interferon-stimulated proteins shown to be critically important in the host immune response against viral infections. These proteins confer intrinsic innate immunity to pH-dependent viruses such as influenza viruses and can inhibit cytosolic entry of such viruses to limit the severity of infection following interferon upregulation. Mutations in IFITM genes in humans have been identified as key risk factors for worsened disease progression, particularly in the case of avian influenza viruses such as H7N9. While the IFITM genes of humans and mice have been well characterized, no studies have been conducted to classify the IFITM locus and interferon-driven upregulation of IFITMs in ferrets. Here, we show the architecture of the ferret IFITM locus and its synteny to the IFITM locus of other mammalian and avian species. Furthermore, we show that ferret IFITM1, -2, and -3 are functionally responsive to both interferon-α (IFN-α) and influenza virus stimulation. Thus, we show that ferret IFITMs exhibit interferon-stimulated properties similar to those shown in other species, furthering our knowledge of the innate immune response in the ferret model of human influenza virus infections. IMPORTANCE IFITM proteins can prevent the entry of several pH-dependent viruses, including high-consequence viruses such as HIV, influenza viruses, and SARS-coronaviruses. Mutations in these genes have been associated with worsened disease outcomes with mutations in their IFITM genes, highlighting these genes as potential disease risk factors. Ferrets provide a valuable tool to model infectious diseases; however, there is a critical shortage of information regarding their interferon-stimulated genes. We identified the putative ferret IFITM genes and mapped their complete gene locus. Thus, our study fills a critical gap in knowledge and supports the further use of the ferret model to explore the importance of IFITMs in these important diseases.

The ferret is a key animal model for investigating the pathogenicity and transmissibility of important human viruses, and for the pre‐clinical assessment of vaccines. However, relatively little is known about the ferret immune system, due in part to a paucity of ferret‐reactive reagents. In particular, T follicular helper (Tfh) cells are critical in the generation of effective humoral responses in humans, mice and other animal models but to date it has not been possible to identify Tfh in ferrets. Here, we describe the screening and development of ferret-reactive BCL6, CXCR5 and PD-1 monoclonal antibodies. We found two commercial anti-BCL6 antibodies (clone K112-91 and clone IG191E/A8) had cross-reactivity with lymph node cells from influenza-infected ferrets. We next developed two murine monoclonal antibodies against ferret CXCR5 (clone feX5-C05) and PD-1 (clone fePD-CL1) using a single B cell PCR-based method. We were able to clearly identify Tfh cells in lymph nodes from influenza infected ferrets using these antibodies. The development of ferret Tfh marker antibodies and the identification of ferret Tfh cells will assist the evaluation of vaccine-induced Tfh responses in the ferret model and the design of novel vaccines against the infection of influenza and other viruses, including SARS-CoV2.

The current pandemic has highlighted the ever-increasing risk of human to human spread of zoonotic pathogens. A number of medically-relevant zoonotic pathogens are negative-strand RNA viruses (NSVs). NSVs are derived from different virus families. Examples like Ebola are known for causing severe symptoms and high mortality rates. Some, like influenza, are known for their ease of person-to-person transmission and lack of pre-existing immunity, enabling rapid spread across many countries around the globe. Containment of outbreaks of NSVs can be difficult owing to their unpredictability and the absence of effective control measures, such as vaccines and antiviral therapeutics. In addition, there remains a lack of essential knowledge of the host–pathogen response that are induced by NSVs, particularly of the immune responses that provide protection. Vaccines are the most effective method for preventing infectious diseases. In fact, in the event of a pandemic, appropriate vaccine design and speed of vaccine supply is the most critical factor in protecting the population, as vaccination is the only sustainable defense. Vaccines need to be safe, efficient, and cost-effective, which is influenced by our understanding of the host–pathogen interface. Additionally, some of the major challenges of vaccines are the establishment of a long-lasting immunity offering cross protection to emerging strains. Although many NSVs are controlled through immunisations, for some, vaccine design has failed or efficacy has proven unreliable. The key behind designing a successful vaccine is understanding the host–pathogen interaction and the host immune response towards NSVs. In this paper, we review the recent research in vaccine design against NSVs and explore the immune responses induced by these viruses. The generation of a robust and integrated approach to development capability and vaccine manufacture can collaboratively support the management of outbreaking NSV disease health risks.

Salmonella enterica serovar Weltevreden is a recently emerged pathogen, and as such we lack a comprehensive knowledge of its microbiology, genomics, epidemiology and biogeography. In this study, we analyzed 174 novel S . Weltevreden isolates including 111 isolates recovered from diarrheal patients in China between 2006 and 2017. Our results demonstrate that the ST365 clone was the predominant causative agent of the diarrhea-outbreak during this period, as vast majority of the isolates recovered from diarrheal patients belonged to this sequence type (97.37%, 74/76). We also determined the ST365 clone as the predominant sequence type of S . Weltevreden from diarrheal patients globally from previously published sequences (97.51%, 196/201). In order to determine the possible antimicrobial genes and virulence factors associated with S . Weltevreden, we performed whole genome sequencing on our novel isolates. We were able to identify a range of key virulence factors associated with S . Weltevreden that are likely to be beneficial to their fitness and pathogenesis. Furthermore, we were able to isolate a novel 100.03-kb IncFII(S) type virulence plasmid that used the same replicon as pSPCV virulence plasmid. Importantly, we demonstrated through plasmid elimination a functional role for this plasmid in bacterial virulence. These findings are critical to further our knowledge of this high consequence pathogen. Importance Salmonella Weltevreden is a newly emerged foodborne pathogen and has caused several outbreaks of diarrheal diseases in some regions in the world. However, comprehensive knowledge of microbiology, genomics, epidemiology and biogeography of this newly emerged pathogen is still lack. In this study, we made an unexpected discovery that S . Weltevreden sequence type (ST) 365 is the causative agent in the diarrhea-outbreak in China and many other regions of the world. We also shown that this sequence type was widely recovered from animal, food, and environmental samples collected in different regions in the world. Importantly, we discovered a novel IncFII(S) type virulence plasmid commonly carried by S . Weltevreden strains of both human, animal, and food origins. These data facilitate future studies investigating the emergence of S . Weltevreden involved in diarrheal outbreaks and the global spread of S . Weltevreden strains.