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Purpose This study aims to address the issue of high-precision measurement of AC electric field. An electro-optical sensor with high sensitivity is proposed for this purpose. Design/methodology/approach The proposed sensor combines electromagnetic induction and fiber Bragg grating (FBG) sensing techniques. It is composed of a sensing probe, a piece or stack of piezoelectric ceramics (PZT) and an FBG. A signal processing circuit is designed to rectify and amplify the induced voltage. The processed signal is applied to the PZT and the deformation of PZT is detected by FBG. Theoretical calculation and simulation are conducted to verify the working principle of the probe. The sensor prototype is fabricated and its performance is tested. Findings The results of this study show that the sensor has good linearity and repeatability. The sensor sensitivity is 0.061 pm/Vm−1 in the range from 250 to 17,500 V/m, enabling a measurement resolution of electric field strength of 16.3 V/m. The PZT stack is used to enhance the sensor sensitivity and the resolution can be improved up to 3.15 V/m. Originality/value A flexure hinge lever mechanism is used to amplify the deformation of PZT for further enhancement of sensitivity. The results show that the proposed sensor has high sensitivity and can be used for the accurate measurement of an electric field. The proposed sensor could have potential use for electric field measurement in the power industry.

S . Enteritidis is a highly adapted pathogen that causes significant economic losses in the poultry industry around the world.

Salmonella enterica serovar Enteritidis (

Salmonella spp. is a major foodborne pathogen that is distributed among most pork production chains worldwide. This study aimed to investigate the dynamic changes in Salmonella spp. along the pig breeding process monthly from April 2018 to March 2019 in a pig farm in Shanghai, China, and identify the potential critical control points during the production. In total, 239 Salmonella spp. isolates were obtained from 1389 samples, in which Salmonella were detected from 26.3% (222/843) of fecal samples, 7.1% (17/240) of feed samples, and 0.0% (0/306) of both water and insect samples. Seven different serotypes were identified, with the predominant serotype being Salmonella Derby (21.8%), followed by Salmonella Typhimurium (18.8%), Salmonella Rissen (16.3%), Salmonella Mbandaka (12.6%), and Salmonella 1,4,[5],12:i:- (11.8%). Most probable number (MPN) analysis revealed that the load of Salmonella spp. gradually increased along the pig production chain, while the highest number of Salmonella spp. isolates was at the fattening stage (MPN value, 11-15 MPN/g). The pulsed-field gel electrophoresis showed that both Salmonella Typhimurium and Salmonella Derby isolates were grouped to six clusters. The antimicrobial resistance analyzed demonstrated that 80.0% of the isolates were of multidrug resistance and resistant to sulfamethoxazole (84.5%), lincomycin (89.4%), ampicillin (96.9%), oxytetracycline (93.8%), and tetracycline (95.1%). We further evaluated the Salmonella spp. Resistance to quaternary ammonium compounds (QACs) showed an increasing trend along with the testing period indicating that the use of QACs could induce the resistance of Salmonella spp. to QACs. Our study confirmed the dynamic changes in Salmonella spp. over time and space in this pig farm and identified feed and the fattening house as the key points for the prevention and control of Salmonella spp. contamination.

Inflammasome activation is an essential innate immune defense mechanism against Salmonella infections. Salmonella has developed multiple strategies to avoid or delay inflammasome activation, which may be required for long-term bacterial persistence. However, the mechanisms by which Salmonella evades host immune defenses are still not well understood. In this study, Salmonella Enteritidis (SE) random insertion transposon library was screened to identify the key factors that affect the inflammasome activation. The type I secretion system (T1SS) protein SiiD was demonstrated to repress the NLRP3 inflammasome activation during SE infection and was the first to reveal the antagonistic role of T1SS in the inflammasome pathway. SiiD was translocated into host cells and localized in the membrane fraction in a T1SS-dependent and partially T3SS-1-dependent way during SE infection. Subsequently, SiiD was demonstrated to significantly suppress the generation of mitochondrial reactive oxygen species (mtROS), thus repressing ASC oligomerization to form pyroptosomes, and impairing the NLRP3 dependent Caspase-1 activation and IL-1β secretion. Importantly, SiiD-deficient SE induced stronger gut inflammation in mice and displayed NLRP3-dependent attenuation of the virulence. SiiD-mediated inhibition of NLRP3 inflammasome activation significantly contributed to SE colonization in the infected mice. This study links bacterial T1SS regulation of mtROS-ASC signaling to NLRP3 inflammasome activation and reveals the essential role of T1SS in evading host immune responses.

Lithium-ion (Li-ion) battery becomes a promising energy storage element in the power grid. Temperature monitoring is a key tool to guarantee the safe service and durable life of Li-ion batteries. In this study, an optical fiber sensor was proposed for the temperature monitoring of Li-ion batteries. The proposed sensor consists of a metal ring and a fiber Bragg grating (FBG), which is gloved on the external electrode of the cell. This installation is not only easy to assemble but also no interference to the circuit connection. The results of calibration test on proposed FBG sensors show good linearity (0.99) and high sensitivity (12 pm/°C). A comparison test illustrates the response of proposed FBG sensors has a good agreement with PT100 sensors. The FBG sensor can be fabricated as sensor arrays to monitor the temperature of multiple cells during charge/discharge cycles, simultaneously. The experimental results show that the FBG sensors have good repeatability during several cycles. FBG sensor arrays can be used to achieve distributed temperature measurement for multiple cells. It provides a solution to monitor the thermal behavior of each cell in battery packs and an available tool to improve the thermal management and safety of Li-ion batteries.

Salmonella is the most important foodborne pathogen in poultry production systems and can infect humans via consumption of contaminated food. Ducks, an important waterfowl widely raised in China, are also a vehicle that transmits Salmonella through the food supply chain. In this study, 701 samples were collected from each production stage of the duck production chain. Salmonella was isolated and identified, and the isolates were tested for drug sensitivity and molecular typing based on whole genome sequencing (WGS) to explore the prevalence of Salmonella in the duck production chain. Altogether, a total of 180 Salmonella isolates (25.7%) were obtained from the duck production chain, 82 (35.7%) isolates were from hatchery samples, followed by 64 (29.2%) from market samples, 17 (23.6%) from farm samples, and 17 (9.4%) from slaughterhouse samples. All isolates were divided into 9 serotypes, among which S. Typhimurium, S. Anatum, and S. Enteritidis were the dominant serotypes. The S. Typhimurium was distributed in various production stages in the duck production chain. Among the 16 antibiotics, selected 60 isolates were only resistant to NAL, indicating that resistance of Salmonella in the duck production chain was low. WGS phylogenetic relationship results based on core-genome SNPs showed that S. Typhimurium can spread across geographic regions and along between different stages of the duck production chain, eventually reaching the market where it is a potential threat to consumer health. This study explored the prevalence of Salmonella in the duck production chain which will provide data support for proposing some interventions to control Salmonella.

Inflammation is a double-edged sword that can be induced by various PAMPs, resulting in the control of infection by invading pathogens or injuries. The inflammatory response requires strict and precise control and regulation. MicroRNAs (miRNAs) are small non-coding RNA molecules that regulate gene expression via translational inhibition or mRNA degradation. However, the role of miRNAs in inflammation induced by flagellin (ligand of TLR5) has yet to be fully determined. In this study, we identified differentially expressed miRNAs in murine bone marrow-derived dendritic cells (BMDCs) between flagellin treatment and medium alone using miRNA microarray. We found that flagellin stimulation downregulated miR-5112 expression in BMDCs and spleen DCs in vitro and in vivo. The overexpression of miR-5112 decreased inflammatory cytokine production, accompanied by a reduction of IKKγ in flagellin-stimulated BMDCs. We demonstrated that miR-5112 could directly target IKKγ to inhibit inflammatory cytokine production. Furthermore, miR-5112 inhibited the inflammatory response induced by flagellin or Salmonella infection in vivo. Interestingly, miR-5112 could also dampen the inflammatory response and alleviate dextran sulfate sodium (DSS)-induced colitis in C57BL/6 mice. These results suggest that miR-5112 could be a novel therapeutic target for both bacterial infection and DSS-induced colitis model.

Egg safety and quality is a long-standing concern for producers and consumers. Salmonella enterica serovar Enteritidis (Salmonella Enteritidis) is the prevalent egg-product-related foodborne pathogen. The treatment with organic acids, including lactic acid, for the elimination and control of foodborne pathogens is generally recognized as safe, and is widely applied in many foods. In this study, we found that spraying with 2% lactic acid on the eggshell surface effectively reduced Salmonella Enteritidis counts. Agar-filled eggs were used to study the bacterial penetration of the eggshell (eggshell and membranes), and the contamination of the whole egg contents was investigated. No significant differences were observed between the Salmonella Enteritidis penetration in lactic acid sprayed and unsprayed eggs stored at 4 °C. The Haugh unit, albumin pH, and eggshell strength values were similar in the sprayed and washed eggs. Average Haugh unit scores for sprayed eggs were still Grade A within 3 weeks. However, the lactic acid treatment decreased the eggshell strength and cuticle layer thickness. SEM observations confirmed that spraying of lactic acid damaged the cuticle integrity. The results from our research will indicate that lactic acid spray is highly effective in reducing Salmonella contamination on the eggshell surface, but trans-shell penetration was demonstrated in the eggs stored at room temperature. Consequently, low-temperature storage is important for the prevention of recontamination of the egg after spraying.

Salmonella is considered one of the most important foodborne pathogens and is commonly associated with the consumption of pork and other animal products. Multidrug resistant (MDR) Salmonella strains are highly adaptive and have been responsible for foodborne disease outbreaks. We investigated Salmonella prevalence and antimicrobial resistance in pork obtained from open markets in Xuzhou, Jiangsu Province, China. From October 2016 to September 2017, a total of 155 Salmonella isolates were recovered from 324 pork samples in different open markets, representing 35.5% of the samples tested. All of the Salmonella isolates were characterized using serotyping, antimicrobial resistance detection, and multilocus sequence typing (MLST). Thirteen serotypes were identified among the 155 Salmonella enterica isolates. S. Derby (28.7%) was the most prevalent serotype, but others, such as S. Enteritidis (18.3%), S. Meleagridis (15.7%), and S. Typhimurium (13.0%), were also widespread. MLST analysis showed that 14 sequence type (ST) patterns were identified and ST40 occupied an absolute superiority among these isolates. Tests of susceptibility to 16 antimicrobial agents using a disk diffusion assay showed that 97.4% of the isolates were resistant to at least one compound, and 66.1% exhibited multidrug resistance, as they were resistant to at least four antimicrobials. The presence of most of the antimicrobial resistance genes was consistent with the resistance phenotypes. Of the antimicrobial resistance genes examined in this study, parC, gyrA, aadA1, sul1, floR, and tetA were the most prevalent resistance genes in the MDR isolates. Our findings show the prevalence of Salmonella in pork in the open markets of Xuzhou, China. Additionally, many Salmonella isolates were resistant to multiple antimicrobial agents, which may aid the optimization of hazard analysis in pork sold in open markets.

H7N9 influenza virus has an unusually high fatality rate of approximately 40%, and a safe and effective vaccine against this subtype is urgently needed. Flagellin, a Toll-like receptor (TLR) 5 agonist, has been deemed as a potent adjuvant candidate. However, its high antigenicity and potential for causing inflammatory injury might restrict its clinical application. Previously, we demonstrated that a fusion protein, HA1-2-FliC, comprising the hemagglutinin globular head protein (HA1-2) of H7N9 influenza virus and the full-length Salmonella typhimurium flagellin protein (FliC), had high efficiency against H7N9 in mouse and chicken models. Here, we constructed an improved fusion protein, HA1-2-FliCΔD2D3, with HA1-2 fused to the FliCΔD2D3 (lacking the hypervariable-region domains D2 and D3 of FliC). HA1-2-FliCΔD2D3 exhibited efficient immunoreactivity and TLR5 agonist efficacy, and promoted innate immune-response activation in mouse macrophages, peripheral blood mononuclear cells, and splenocytes, based on cytokine- and chemokine-expression profiles. Mice immunized with HA1-2-FliCΔD2D3 showed significantly lower systemic inflammatory responses (compared with HA1-2-FliC) and highly reduced flagellin-specific antibody production, without affecting HA1-2-specific antibody production and cellular immune responses. Enhanced IFN-γ/IL-4 generation suggested that HA1-2-FliCΔD2D3 maintained balanced Th1/Th2 immune responses. Furthermore, virus challenge was performed in a chicken model. The results showed that chickens receiving FliCΔD2D3 adjuvant vaccine induced high levels of serum neutralizing antibodies, and exhibited a significant reduction of viral loads in throat and cloaca compared to chickens receiving only HA1-2. In conclusion, we constructed the H7N9 influenza subunit vaccine candidate HA1-2-FliCΔD2D3, with reduced immunogenicity against FliC and lower adverse events. The improved adjuvant FliCΔD2D3 can potentially help in developing safe and effective universal protein-based influenza vaccines for humans.

Multisensory and artificial intelligence approaches are key tools to achieve intelligent management of future battery systems. Strain monitoring using optical fiber sensors is an important role of multi-sensing in batteries. In this paper, the strain of batteries is monitored by fiber Bragg grating sensors, and the strain data are used to estimate the state of charge (SoC) and state of health (SoH) of batteries. A Kalman filtering (KF) model is proposed for SoC estimation based on strain signal of cells. Moreover, this work employs an artificial neural network (NN) for SoC estimation based on the strain data. The experimental data are acquired from commercial lithium-ion cells under two operating conditions. The KF model is established based on multiple regression between strain and SoC, which shows good performance in estimation for the static cycles. For NN estimators, input variables with strain parameter can enhance the accuracy of SoC estimation. A KF model based on the peak strain is developed to estimate the capacity degradation of battery, and the results show that strain can be used as an indicator to estimate SoH. The results present an encouraging outcome that SoC estimation can be achieved using non-electrical parameters solely, and the strain signal can also be used as an auxiliary parameter to improve the accuracy of SoC estimation. This new exploration provides a basis for multi-parameter cooperative estimation of battery state in the future battery system with a multisensory approach. This work was supported by the National Natural Science Foundation of China (52075429 and 92060110) , Natural Science Foundation of Jiangxi Province (20202BAB204019).

Microbial infection can trigger the assembly of inflammasomes and promote secretion of cytokines, such as IL-1β and IL-18. It is well-known that Salmonella modulates the activation of NLRC4 (NLR family CARD domain-containing protein 4) and NLRP3 (NLR family pyrin domain-containing 3) inflammasomes, however the mechanisms whereby Salmonella avoids or delays inflammasome activation remain largely unknown. Therefore, we used Salmonella Enteritidis C50336ΔfliC transposon library to screen for genes involved in modulating inflammasomes activation. The screen revealed the galactose metabolism-related gene galE to be essential for inflammasome activation. Here, we found that inflammasome activation was significantly increased in J774A.1 cells or wild-type bone marrow-derived macrophages (BMDMs) during infection by ΔfliCΔgalE compared to cells infected with ΔfliC. Importantly, we found that secretion of IL-1β was Caspase-1-dependent, consistent with canonical NLRP3 inflammasome activation. Furthermore, the virulence of ΔfliCΔgalE was significantly decreased compared to ΔfliC in a mouse model. Finally, RNA-seq analysis showed that multiple signaling pathways related to the inflammasome were subject to regulation by GalE. Taken together, our results suggest that GalE plays an important role in the regulatory network of Salmonella evasion of inflammasome activation.

Salmonella enterica serovar Enteritidis (S. Enteritidis) is an important zoonotic pathogen that can lead to diarrhea and systemic infections in humans and mortality in animals. This is a major public health issue worldwide. Safe and effective vaccines are urgently needed to control and prevent Salmonella infection. Subunit vaccines are safe and provide targeted protection against Salmonella spp. Here, we developed and evaluated an S. Enteritidis subunit vaccine candidate, the rHis-SseB adjuvant with simvastatin. We amplified the SseB gene from S. Enteritidis C50041 genomic DNA and expressed the recombinant proteins rHis-SseB and rGST-SseB using the Escherichia coli system. Western blotting confirmed the immunoreactivity of recombinant proteins rHis-SseB and rGST-SseB with antisera against Salmonella Enteritidis C50041. In a mouse model of intramuscular vaccination, co-immunization with rHis-SseB and simvastatin significantly enhanced both the SseB-specific antibody titer in serum (humoral immune response) and splenic lymphocyte proliferation (cellular immune response). Co-immunization with rHis-SseB and simvastatin provided 60% protection against subsequent challenge with the S. Enteritidis C50041 strain and decreased bacterial colonization in the liver and spleen. These findings provide a basis for the development of an S. Enteritidis subunit vaccine.

Salmonella is recognized as a major human foodborne pathogen and threat to public health world widely. It is important to carry out epidemiological investigations to determine the primary sources of bacterial contamination. Pulsed-field gel electrophoresis (PFGE) is an important method of the molecular typing, and play an important role in tracking the sources of infection and epidemic control. The PFGE is currently considered as "gold standard" of molecular typing methods for bacterial foodborne pathogen. Here, we describe the PFGE protocol to type the Salmonella from pork.

Salmonella is recognized as a major human foodborne pathogen and threat to public health world widely. It is important to carry out epidemiological investigations to determine the primary sources of bacterial contamination. Pulsed-field gel electrophoresis (PFGE) is an important method of the molecular typing, and play an important role in tracking the sources of infection and epidemic control. The PFGE is currently considered as "gold standard" of molecular typing methods for bacterial foodborne pathogen. Here, we describe the PFGE protocol to type the Salmonella from pork.

L ive a ttenuated S almonella v accine (LASV) is considered to be an effective contributory measure during the control of Salmonella infection. A Salmonella Pullorum spiC mutant was evaluated comprehensively as a LASV candidate (LASV-p) for broilers in terms of safety and immunogenicity. LASV-p was adminstered to 3-day broilers by intramuscular injection. The LD50 increased 126 fold, and no tissue lesions were observed in the liver, spleen and cecum, in comparison with the control group inoculated with PBS and a passive group by wild-type Salmonella. Growth rates of all broilers were normal and not affected. LASV-p persisted in vivo until 21 days in liver, 28 days in spleen and 35 days in feces, and induced high levels of humoral IgG and mucosal IgA. Cellular immunity was also stimulated in the form of antigen-specific lymphocyte proliferation and higher counts of CD3+CD8+ T cells and increased expression of mRNA of Th1 cytokines, IFN-γ and IL-2, in the early stage, and Th2 cytokines, IL-4 and IL-10, in the later stages. LASV-p provided at least 90% immuneprotection against a wild-type Salmonella Pullorum and cross-protection in different degree against other Salmonella searovars. Oral vaccine could also offer high immune protection of 87.5%. These results indicated that LASV-p vaccine candidate had a high level of safety and immune protection and it might be developed as a novel easy-to-use oral vaccine to improve poultry health in the future.

Background Salmonella Enteritidis (SE) is one of the major foodborne zoonotic pathogens of worldwide importance which can induce activation of NLRC4 and NLRP3 inflammasomes during infection. Given that the inflammasomes play an essential role in resisting bacterial infection, Salmonella has evolved various strategies to regulate activation of the inflammasome, most of which largely remain unclear. Results A transposon mutant library in SE strain C50336 was screened for the identification of the potential factors that regulate inflammasome activation. We found that T3SS-associated genes invC, prgH, and spaN were required for inflammasome activation in vitro. Interestingly, C50336 strains with deletion or overexpression of Dam were both defective in activation of caspase-1, secretion of IL-1β and phosphorylation of c-Jun N-terminal kinase (Jnk). Transcriptome sequencing (RNA-seq) results showed that most of the differentially expressed genes and enriched KEGG pathways between the C50336-VS-C50336Δdam and C50336-VS-C50336::dam groups overlapped, which includes multiple signaling pathways related to the inflammasome. C50336Δdam and C50336::dam were both found to be defective in suppressing the expression of several anti-inflammasome factors. Moreover, overexpression of Dam in macrophages by lentiviral infection could specifically enhance the activation of NLRP3 inflammasome independently via promoting the Jnk pathway. Conclusions These data indicated that Dam was essential for modulating inflammasome activation during SE infection, there were complex and dynamic interplays between Dam and the inflammasome under different conditions. New insights were provided about the battle between SE and host innate immunological mechanisms.

Salmonella has been known as the most important foodborne pathogen, which can infect humans via consuming contaminated food. Chicken meat has been known as an important vehicle to transmit Salmonella by the food supply chain. This study determined the prevalence, antimicrobial resistance, and genetic characteristics of Salmonella at different chicken slaughtering stages in East China. In total, 114 out of 200 (57%) samples were Salmonella positive, while Salmonella contamination was gradually increasing from the scalding and unhairing stage (17.5%) to the subdividing stage (70%) throughout the slaughtering. Whole-genome sequencing (WGS) was then performed to analyze the serotype, antimicrobial resistance gene profiles, and genetic relationship of all Salmonella isolates. The most common serotypes were S. Kentucky (51/114, 44.7%) and S. Enteritidis (37/114, 32.5%), which were distributed throughout the four slaughtering stages, and were also identified in the corresponding environments. The multilocus sequence typing (MLST) analysis revealed that seven sequence types (STs) were occupied by six different serotypes, respectively. Only S. Kentucky had two STs, ST314 was the predominant ST shared by 50 isolates, while the ST198 has 1 isolate. The antimicrobial resistance gene analysis demonstrated that most of the strains belonging to S. Kentucky (39/51, 76.5%) and S. Indiana (15, 100%) contained over five groups of antimicrobial resistance genes. Based on the core genome analysis, 50 S. Kentucky isolates were genetically identical, indicating that one S. Kentucky strain with the same genetic background was prevalent in the chicken slaughtering line. Although 37 S. Enteritidis isolates only had three different antimicrobial resistance gene profiles, the core genome sequence analysis subtyped these S. Enteritidis isolates into five different clusters, which revealed the diverse genetic background of S. Enteritidis in the slaughterhouse. The antimicrobial resistance phenotypes were consistent with the presence of the corresponding resistance genes of S. Kentucky and S. Enteritidis, including tetA, floR, blaTEM-1B, strA/B, sul1/sul2, and gyrA (D87Y). Our study observed a high prevalence of Salmonella in the chicken slaughter line and identified the slaughtering environment as a main source of causing Salmonella cross-contamination during chicken slaughtering. Further studies will be needed to limit the transmission of Salmonella in the slaughterhouse.

Additional file 1: Figure S1. Growth curves of dam, invC, hilD, prgH, and spaN gene deletion mutants, complementation, and overexpression strains. Figure S2. Two rounds of screening to identify the genes involved in regulating inflammasome activation in vitro (original images of immunoblotting). Figure S3. Deletion mutants of dam, invC, prgH, and spaN failed to induce inflammasome activation (original images of immunoblotting). Figure S4. Deletion of prgH did not influence the synthesis of inflammasome components (original images of immunoblotting). Figure S5. Overexpression of Dam inhibited inflammasome activation (original images of immunoblotting). Figure S6. The ability of the dam complementation strain and overexpression strain cultured without IPTG to activate the inflammasome was improved (original images of immunoblotting). Figure S7. The deletion of Dam blocked the Jnk pathway in infected J774A.1 cells (original images of immunoblotting). Figure S8. Dam independently promoted NLRP3 inflammasome activation (original images of immunoblotting).

Salmonella enterica serovar Enteritidis (S. Enteritidis, SE) is a major cause of foodborne diseases for humans. The completeness of the O-chain antigen of Lipopolysaccharide (LPS) determines whether a S. Enteritidis strain is smooth or rough. However, genes that are involved in the synthesis of LPS and rough-smooth variation are not completely understood. In this study, we used monoclonal antibody against O-antigens (O9 mAb) to identify novel factors that are involved in LPS synthesis and rough variation in S. Enteritidis by using signature-tagged mutagenesis (STM) technique. Our results showed that transposon insertion in the gene rfbH led to different LPS phenotype, auto-aggregation characteristic, motility and resistance to environmental stress compared with SE wild-type strain C50041. In addition, sera tests showed that rfbH mutant does not elicit specific antibodies against O-antigens in vaccinated animals. Taken together, the S. Enteritidis rfbH gene is implicated in LPS biosynthesis, rough variation, sera distinguishable reaction, motility and stress resistance. The rfbH mutant strain could be potentially used as a distinguishable vaccine or a live vector to deliver drugs and antibodies in vivo.

With the development and application of portable devices and electronic products, the problem of cooling is facing severe challenge. However, traditional compression type refrigeration technology cannot meet the current demand, and the electrocaloric (EC) cooling technology was considered as the most promising choice. In this paper, BST nanoparticles, BN nanosheets and P(VDF-TrFE-CFE) were blended to prepare a uniformly dispersed nanocomposite, which not only increased the polarization, but also improved the breakdown strength and thermal conductivity of the material effectively. Then, a EC cooling device driven by electromagnetic was manufactured. The average cooling heat flux of the EC cooling device can reach 293.14 W/m2 under electric field of 40 MV/m at 1 Hz. The specific cooling power of the EC cooling device can reach 0.96 W/g, and the coefficient of performance (COP) is 3.6. Finally, the EC cooling device performs well in practical applications, the temperature drop can reach 12 °C within 20 s. Compared with the traditional fan cooling, the efficiency was improved significantly. This research achieves efficient electrocaloric cooling through the composite of ferroelectric materials, but also paves the way for the application of EC cooling technology in portable devices and electronic products.

Waste heat is very abundant and widespread in our lives. Using pyroelectric generators (PEG) to capture the ambient heat energy is meaningful. Here we fabricated a highly efficient PEG using nanocomposite. The nanocomposite was made of P(VDF-TrFE-CFE) and barium strontium titanate (BST) nanoparticles and boron nitride (BN) nanosheets, which has excellent thermal conductivity and high polarization. The output current density of PEG can reach 1.7 μA/cm2, it is a great improvement for PEG devices. In addition, we present a new evaluation method for standardizing the performance of a PEG, which is use η (the ratio between the power density and the heat flux) as the thermos-electric conversion efficiency of PEG. Furthermore, a PEG-unit was designed and fabricated, which was cleverly connected by PEG and a plane spiral spring, it can better utilize external vibration to realize waste heat recovery without auxiliary device. The PEG-unit can be used for energy recovery in the process of train running and it can light up the LED lamp bead for 10 s within five minutes operation.

TNF receptor-associated factor 6 (TRAF6) is an adaptor protein and an E3 ubiquitin ligase mediating multiple cell signaling pathway activation in a context-dependent manner. TRAF6 plays critical roles in innate immune response and regulates function of antigen-presenting cells. Here, we cloned the goose TRAF6 (goTRAF6) gene from a healthy Yangzhou great white goose (Anser anser), which had a typical TRAF structure and shared a high-sequence identity with TRAF6 of other birds. Quantitative real-time PCR revealed that goTRAF6 mRNA was broadly expressed in all the studied tissues, with highest expression in the heart and pectoral muscle. Overexpression of goTRAF6 caused NF-κB activation in a dose-dependent manner and substantially upregulated IFN-β expression in HEK293T cells. Following Toll-like receptor (TLR) ligand stimulation of goose peripheral blood mononuclear cells, goTRAF6 and downstream inflammatory cytokine mRNA levels considerably up-regulated, especially at early stages. Salmonella Enteritidis challenge caused overexpression of goTRAF6 and cytokine mRNA in all the examined organs. These findings demonstrated that goTRAF6 played a substantial role in TLR-TRAF6 signaling cascade, and further contributed to the antibacterial-responses in host.

This study surveyed the distribution, antimicrobial susceptibility profiles and serotype of Salmonella isolated from three slaughterhouses, and performed molecular typing on these isolates, to understand the relationship between strains of Salmonella obtained from the pork production chain in Yangzhou, China. Samples from slaughtered pigs and the slaughtering environment were collected from three slaughterhouses in Yangzhou, Jiangsu province, from October 2012 to July 2013. The positive identification rates of Salmonella in slaughtered pigs and the environmental samples were 46.6% and 48.8%, respectively. The prevalence of Salmonella in slaughterhouses were affected by seasonal factors and reached the peak in summer. Among the Salmonella serovars identified, S. Derby was most prevalent in slaughterhouses, but other serovars like S. Typhimurium, S. Meleagridis and S. Anatum were also widespread. Antimicrobial susceptibility testing revealed that 32 and 131 different MDR patterns were found among the strains from the environment and slaughtered pig samples, respectively. Fifty-six isolates of S. Derby and 16 strains of S. Typhimurium were characterized by the technique of pulsed-field gel electrophoresis (PFGE) using the restriction enzyme Xba I. 35 and 11 PFGE patterns were generated among the selected isolates. Four isolates of S. Derby isolates with the same pattern (PF26) were isolated from cooling water, evisceration and carcass, suggesting that cross contamination occurred between the environment and the slaughtered pigs. Six S. Typhimurium in cluster 1 with the same ST type (ST19) came from different parts of the slaughtered pig, which could have occurred because of horizontal transmissions along the slaughtering process. The same PFGE patterns of Salmonella were found in both samples from carcasses in the slaughterhouse and in the Yangzhou pork market, proving that Salmonella had spread from the slaughterhouse to the pork market. In conclusion, our study demonstrate that serious cross contamination occurred in Yangzhou slaughterhouses and can contribute Salmonella contamination in pork sold in the local public market.

Salmonella Enteritidis (SE) is a highly adapted pathogen causing severe economic losses in the poultry industry worldwide. Chickens infected by SE are a major source of human food poisoning. Vaccination is an effective approach to control SE infections. This study evaluated the immunogenicity and protective efficacy of a SE sptP deletion mutant (C50336ΔsptP) as a live attenuated vaccine (LAV) candidate in chickens. 14 day-old specific pathogen-free (SPF) chickens were intramuscularly immunized with various doses of C50336ΔsptP. Several groups of chickens were challenged with the virulent wild-type SE strain Z-11 via the same route at 14 days post vaccination. Compared to the control group, the groups vaccinated with 1 × 106, 1 × 107 and 1 × 108 colony-forming units (CFU) of C50336ΔsptP exhibited no clinical symptoms after immunization. Only slight pathological changes occurred in the organs of the 1 × 109 CFU vaccinated group. C50336ΔsptP bacteria were cleared from the organs of immunized chickens within 14 days after vaccination. Lymphocyte proliferation and serum cytokine analyses indicated that significant cellular immune responses were induced after the vaccination of C50336ΔsptP. Compared to the control group, specific IgG antibody levels increased significantly in vaccinated chickens, and the levels increased markedly after the challenge. The 1 × 107, 1 × 108, and 1 × 109 CFU vaccinated chickens groups showed no clinical symptoms or pathological changes, and no death after the lethal challenge. Whereas severe clinical signs of disease and pathological changes were observed in the control group chickens after the challenge. These results suggest that a single dose of C50336ΔsptP could be an effective LAV candidate to against SE infection in chickens.

Salmonella is commonly isolated from raw pork and is a leading cause of foodborne illness. Because China has the highest rate of pork consumption and the largest number of pig breeding facilities in the world, an epidemiological analysis of Salmonella species from pork in China is warranted. In this study, pork samples (n = 1,096) were collected from 20 major free markets in four cities of Jiangsu province from August 2010 to December 2012. A total of 163 Salmonella isolates were recovered from 154 Salmonella-positive samples. Among 14 Salmonella serovars identified, Derby (47.9%) was most prevalent, followed by Typhimurium (10.4%), Meleagridis (9.2%), Anatum (8.6%), and London (6.7%). Antimicrobial sensitivity testing revealed that 134 (82.2%) of the isolates were resistant to at least one antimicrobial agent, and 41 (25.2%) were resistant to more than three antimicrobials. The highest resistance was to tetracycline (66.3% of isolates) followed by ampicillin (39.9%), trimethoprim-sulfamethoxazole (31.3%), and nalidixic acid (30.1%). Multilocus sequence typing analysis revealed 14 sequence type (ST) patterns; ST40 was the most common (77 isolates) followed by ST64 (19 isolates). Our research revealed a high prevalence of Salmonella in retail pork. Diversity among the Salmonella isolates was high in terms of serovar and genotype, and multidrug resistance was prevalent. Multilocus sequence type was generally associated with serovar and provided a reliable prediction of the most common Salmonella serovars.

Summary: Toll-like receptors (TLRs) activate innate immunity via interactions between their Toll/interleukin-1 (IL-1) receptor (TIR) domain and downstream adaptor proteins. Here we report that Salmonella Enteritidis produces a secreted protein (TcpS) that contains both a TIR domain and a coiled-coil domain. TcpS blocks MyD88- and TRIF-mediated TLR signaling, inhibits inflammatory responses, and promotes bacterial survival. Early-stage immune evasion by TcpS results in severe tissue damage in the late stage of infection and contributes to Salmonella virulence. TcpS-derived peptides inhibit nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) activation and reduce lipopolysaccharide (LPS)-elicited systemic inflammation. Therapeutic peptide administration alleviates weight loss of mice infected with H1N1 influenza. Importantly, maximal TcpS-mediated TLR inhibition requires the critical TIR-TcpS residues Y191 and I284, as well as TcpS homodimerization via its N-terminal coiled-coil domain. Our study unveils a mechanism in which TcpS suppresses innate immunity via both its homodimerization and interaction with MyD88. TcpS is also a potential therapeutic agent for inflammation-associated diseases. : Xiong et al. show that Salmonella subverts host TLR signaling by delivering a protein containing a TIR domain and a coiled-coil domain, TcpS. Immune evasion by TcpS supports early bacterial infection, which enables late-stage virulence and inflammation storm. TcpS-derived peptides reduce LPS-elicited systemic inflammation and are protective against influenza challenge in mice. Keywords: Salmonella, coiled-coil, TIR, TcpS, TLR signaling, immune evasion, inflammation, therapeutic peptide

An epidemiological analysis of Salmonella spp. from food animals and humans in eastern China was conducted from 2008 to 2011. A total of 135 Salmonella isolates were recovered from 132 of 2566 tested fecal samples, representing 5.1% positives of the samples tested. Ten serotypes were identified among the 135 Salmonella enterica isolates, with Salmonella ser. Senftenberg, Salmonella ser. Enteritidis, Salmonella ser. Pullorum, and Salmonella ser. Typhimurium being the most common. Forty-seven pulsed-field gel electrophoresis (PFGE) profiles were generated among the 135 isolates, which grouped into six clusters (named A–F respectively). PFGE typing grouped the majority of isolates according to serotype, except for Salmonella ser. Senftenberg, Salmonella ser. Derby, and Salmonella ser. Saintpaul. And, more remarkable, no overlap in the PFGE patterns between the human and animal isolates was found. Antimicrobial testing revealed that 131 (97.0%) isolates were resistant to at least one antimicrobial. The highest prevalence of resistance was to carbenicillin (94.1%), followed by nalidixic acid (45.2%), sulfafurazole (37.0%), ampicillin (31.9%), and amoxicillin (28.9%). Additionally, bla tem-1 (95.3%), bla pse-1 (57.8%), sul2 (78.0%), and tetA (82.4%) were the most prevalent resistance genes, and other than the aminoglycoside resistance genes, the resistance phenotypes were consistent with the identified antimicrobial resistance genes. Finally, class 1 integrons were detected in eight (5.9%) of the 135 isolates, with all of the positive isolates containing int1 and qacEΔ1-sul I . Six positive isolates with a 1 kb amplicon size harbored the aadA2 cassettes. However, class 1 integrons with a 600 bp amplicon did not contain the integron-associated gene cassettes. Our findings showed the prevalence of Salmonella in food animals and humans in eastern China, and that the Salmonella isolates were phenotypically and genetically diverse. Additionally, many Salmonella isolates were resistant to multiple antimicrobial agents, which was consistent with the identified antimicrobial resistance genes.

Background Salmonella enterica serovar Enteritidis (S. Enteritidis) has emerged as one of the most important food-borne pathogens for humans. Lipopolysaccharide (LPS), as a component of the outer membrane, is responsible for the virulence and smooth-to-rough transition in S. Enteritidis. In this study, we screened S. Enteritidis signature-tagged transposon mutant library using monoclonal antibody against somatic O9 antigen (O9 MAb) and O9 factor rabbit antiserum to identify novel genes that are involved in smooth-to-rough transition. Results A total of 480 mutants were screened and one mutant with transposon insertion in rfbG gene had smooth-to-rough transition phenotype. In order to verify the role of rfbG gene, an rfbG insertion or deletion mutant was constructed using λ-Red recombination system. Phenotypic and biological analysis revealed that rfbG insertion or deletion mutants were similar to the wild-type strain in growth rate and biochemical properties, but the swimming motility was reduced. SE Slide Agglutination test and ELISA test showed that rfbG mutants do not stimulate animals to produce agglutinating antibody. In addition, the half-lethal dose (LD50) of the rfbG deletion mutant strain was 106.6 -fold higher than that of the parent strain in a mouse model when injected intraperitoneally. Conclusions These data indicate that the rfbG gene is involved in smooth-to-rough transition, swimming motility and virulence of S. Enteritidis. Furthermore, somatic O-antigen antibody-based approach to screen signature-tagged transposon mutants is feasible to clarify LPS biosynthesis and to find suitable markers in DIVA-vaccine research.

Tumor necrosis factor receptor-associated factor 3 (TRAF3) plays a key antiviral role by promoting type I interferon production. We cloned the pigeon TRAF3 gene (PiTRAF3) according to its predicted mRNA sequence to investigate its function. The 1704-bp full-length open reading frame encodes a 567-amino acid protein. One Ring finger, two TRAF-type Zinc fingers, one Coiled coil, and one MATH domain were inferred. RT-PCR showed that PiTRAF3 was expressed in all tissues, with relatively weak expression in the heart and liver. In HEK293T cells, over-expression of wild-type, △Ring, △Zinc finger, and △Coiled coil PiTRAF3, but not a △MATH form, significantly increased IFN-β promoter activity. Zinc finger and Coiled coil domains were essential for NF-κB activation. In chicken HD11 cells, PiTRAF3 increased IFN-β promoter activity and four domains were all contributing. R848 stimulation of pigeon peripheral blood mononuclear cells and splenocytes significantly increased expression of PiTRAF3 and the inflammatory cytokine genes CCL5, IL-8, and IL-10. These data demonstrate TRAF3's innate immune function and improve understanding of its involvement in poultry antiviral defense.

In 2009, a novel pandemic swine-origin influenza A (H1N1) virus caused a public emergency of international concern. Vaccination is the primary strategy for the control of influenza epidemics. However, the poor immunopotency of many vaccine antigens is a major barrier to the development of effective vaccines against influenza. Flagellin, a Toll-like receptor 5 (TLR5) ligand, has been used as an adjuvant to enhance the immunopotency of vaccines in preclinical studies. Here, we developed a recombinant candidate vaccine, HA1-2-fljB, in which the globular head of the hemagglutinin (HA) antigen (residues 62-284) from H1N1 virus was fused genetically to the N-terminus of Salmonella typhimurium fljB. The recombinant HA1-2-fljB protein was expressed efficiently in Escherichia coli, and the immunogenicity and protective efficacy of recombinant HA1-2-fljB were evaluated in a mouse model. Immunization with HA1-2-fljB elicited robust IgG antibodies and neutralizing antibodies and completely protected the mice against infection by swine-origin influenza A/swine/Jangsu/38/2010 (H1N1). These results suggest that HA antigen placed at the N-terminus of flagellin is also an excellent starting point for creating a fusion HA1-2-fljB protein as a candidate vaccine, and the recombinant HA1-2-fljB protein will contribute to the development of a more effective vaccine against swine-origin influenza virus infection.

Newcastle disease (ND), a highly contagious, acute, and potent infectious disease caused by Newcastle disease virus (NDV), has a considerable impact on the global poultry industry. Although both live attenuated and inactivated vaccines are used to prevent and control the spread of ND among chickens, the increasing number of ND outbreaks in commercial poultry flocks worldwide indicates that routine vaccinations are insufficient to control ND. Hence, efforts are being invested into developing alternative and more effective vaccination strategies. In this study, we focus on F protein, the neutralizing and protective antigen of NDV, and flagellin (FliC), a toll-like receptor 5 (TLR5) agonist that is an effective inducer of innate immune responses. We amplified F gene from velogenic NDV strain F48E8. The recombinant histidine (His)-tagged F protein was efficiently expressed in a Pichia pastoris (P. pastoris) eukaryotic system and verified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. The conditions for F protein expression in P. pastoris were optimal. The immunogenicity of F protein with FliC as the adjuvant was evaluated in a C3H/HeJ mouse model. FliC was found to enhance both F-specific and NDV-specific IgG responses and F-specific cellular immune responses following intraperitoneal co-administration with F protein. Thus, the recombinant F protein expressed by P. pastoris when used with flagellin as the adjuvant has potential as a subunit vaccine candidate.

Toll-like receptor 5 (TLR5) is responsible for the recognition of bacterial flagellin in vertebrates. In this study, we cloned the single-exon TLR5 gene of the Maya breed of Common Shelduck (Tadorna tadorna). The TLR5 open reading frame is 2580 bp in length and encodes an 859-amino acid protein. The putative amino acid sequence of duck TLR5 consisted of a signal peptide sequence, 11 leucine-rich repeat domains, a leucine-rich repeat C-terminal domain, a transmembrane domain, and an intracellular Toll-interleukin-1 receptor domain. The duck TLR5 gene was highly expressed in the lung, bone marrow, spleen, and liver; moderately expressed in kidney, small intestine, large intestine, and brain. A plasmid expressing duck TLR5 was constructed and transfected into HEK293T cells, and expression was confirmed by indirect immunofluorescence assay. HEK293T cells transfected with duck TLR5- and NF-κB-luciferase-containing plasmids significantly responded to flagellin from Salmonella typhimurium, indicating that it is a functional TLR5 homolog.

Toll-like receptor 7 (TLR7) is responsible for recognizing viral single-stranded RNA and antiviral imidazoquinoline compounds, leading to the activation of the innate immune response. In this study, mutated pigeon TLR7 fragments, in which the insertion at position 10 of leucine-rich repeat 10 (LRR10) or at position 15 of LRR2/11/13/14 was deleted, were amplified with an overlap-PCR method, and inserted into the expression vector pCMV. The immune functions of the TLR7 mutants were determined with an NF-κB luciferase assay of transfected cells. The deletion of the insertions absolutely abolished TLR7-NF-κB signaling. With quantitative real-time PCR and sandwich enzyme-linked immunosorbent assay, we observed that stimulation with R848 failed to induce the expression of interleukin 8 (IL-8) in any of the mutant-TLR7-transfected cells, consistent with their lack of NF-κB activity. However, the expression of interferon α (IFN-α) and tumor necrosis factor α (TNF-α) was significantly upregulated in the Del10IN10 and Del14IN15 groups. Remarkably, the levels of pigeon TLR7 expression were significantly increased in all the TLR7-mutated groups. Therefore, we speculate that another part of the deficient TLR7 mediates the induction of IFN-α and TNF-α by increasing the expression of TLR7 as compensation. However, the increased expression of TLR7 in the Del11IN15 group failed to induce the production of IFN-α, IL-8, or TNF-α, indicating that a false compensation occurred when the crucial LRR insertion was deleted.

With an aim to develop a safe, immunogenic fowl typhoid (FT) vaccine, the safety and efficacy of 1009ΔspiCΔcrp, a spiC and crp deletion mutant of Salmonella gallinarum, were evaluated in chickens. Three-day-old chickens were intramuscularly immunized with 1009ΔspiCΔcrp (1×10(7)CFU) and boosted 7days later (at 10-days old) with the same dose and via the same route (vaccinated group). The vaccinated group showed no clinical symptoms and no differences in body weight compared to the unvaccinated control group. 1009ΔspiCΔcrp bacteria colonized and persisted in the liver and spleen of vaccinated chickens for >14days, and significant specific humoral and cellular immune responses were induced. Vaccinated chickens were challenged with S. gallinarum strain SG9 at 21days post-immunization (24-day-old chickens), and efficient protection was observed based on the mortality and clinical symptoms, as compared to those in the control group. These results demonstrate that 1009ΔspiCΔcrp can be used as a live attenuated vaccine.

To develop a novel H5 subtype avian influenza vaccine candidate based on flagellin.HA1-2-fljB was amplified through splicing hemagglutinin gene fragment HA1-2 of A/Goose/Jiangsu/1/2000 H5N1 and Salmonella typhimurium II phase flagellin fljB gene by overlap PCR, and then the gene was inserted to pET32a(+) to construct the recombinant plasmid pET32a-HA1-2-fljB. The recombinant plasmid was transformed into E.coli BL21 (DE3) and was induced with IPTG. The expression of the fusion protein was identified by SDS-PAGE and Western blotting. The biological activity of the fusion protein was evaluated by stimulating HEK293-TLR5 cells.The recombinant plasmid pET32a-HA1-2-fljB was constructed. The fusion protein, whose relative molecule weight was about 100 000, had a good immunoreactivity. TLR5 bioassay demonstrated the flagellin part of the fusion protein could be recognized by TLR5 of HEK293-TLR5 cell line to secrete a higher level of IL-8 when compared with HA1-2 protein (P0.01).The flagellin-based fusion protein HA1-2-fljB with TLR5 bioactivity was expressed correctly, and the results would establish a basis for the further research on avian influenza virus H5 vaccine.

Toll-like receptors (TLRs) are pattern recognition receptors (PRRs) that are vital to activation of the innate immune system in response to invading pathogens through their recognition of pathogen-associated molecular patterns (PAMPs). TLR5 is responsible for the recognition of bacterial flagellin in vertebrates. In this study, we cloned the goose TLR5 gene using rapid amplification of cDNA ends (RACE). The open reading frame (ORF) of goose TLR5 cDNA is 2583 bp in length and encodes an 860 amino acid protein. The entire coding region of the TLR5 gene was successfully amplified from genomic DNA and contained a single exon. The putative amino acid sequence of goose TLR5 consisted of a signal peptide sequence, 11 leucine-rich repeat (LRR) domains, a leucine-rich repeat C-terminal (LRR-CT) domain, a transmembrane domain and an intracellular Toll-interleukin-1 receptor (TIR) domain. The amino acid sequence of goose TLR5 shared 50.5% identity with human (Homo sapiens), 49.8% with mouse (Mus musculus) and 82.7% with chicken (Gallus gallus). The goose TLR5 gene was highly expressed in the spleen, liver and brain; moderately expressed in PBMCs, kidney, lung, heart, bone marrow, small intestine and large intestine; and minimally expressed in the cecum. HEK293 cells transfected with goose TLR5 and NF-κB-luciferase containing plasmids significantly responded to flagellin from Salmonella typhimurium indicating that it is a functional TLR5 homologue. In response to infection with S. enterica serovar Enteritidis (SE), the level of TLR5 mRNA significantly increased over the control in PBMCs at 1 d post infection (p.i.) and was slightly elevated in the spleen at 1 d or 3 d p.i. IL-6 was expressed below control levels in PBMCs but was upregulated in the spleen. In contrast to IL-6, an evident decrease in the expression level of IL-8 was observed in both PBMCs and spleens at 1 d or 3 d p.i. SE challenge also resulted in an increase in the mRNA expression of IL-18 and IFN-γ in PBMCs and the spleen. These results imply that the expression of goose TLR5 is differentially regulated in various tissues and may participate in the immune response against bacterial pathogens.

Recombinant attenuatedSalmonellavaccines have been extensively studied, with a focus on eliciting specific immune responses against foreign antigens. However, very little is known about the innate immune responses, particularly the role of flagellin, in the induction of innate immunity triggered by recombinant attenuatedSalmonellain chickens. In the present report, we describe twoSalmonella entericaserovar Typhimurium vaccine strains, wild-type (WT) or flagellin-deficient (flhD)Salmonella, both expressing the fusion protein (F) gene of Newcastle disease virus. We examined the bacterial load and spatiotemporal kinetics of expression of inflammatory cytokine, chemokine, and Toll-like receptor 5 (TLR5) genes in the cecum, spleen, liver, and heterophils following oral immunization of chickens with the twoSalmonellastrains. TheflhDmutant exhibited an enhanced ability to establish systemic infection compared to the WT. In contrast, the WT strain induced higher levels of interleukin-1β (IL-1β), CXCLi2, and TLR5 mRNAs in cecum, the spleen, and the heterophils than theflhDmutant at different times postinfection. Collectively, the present data reveal a fundamental role of flagellin in the innate immune responses induced by recombinant attenuatedSalmonellavaccines in chickens that should be considered for the rational design of novel vaccines for poultry.