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As an important part of regional coordinated development, the high-quality development of the Yellow River Basin has become a national strategy. It is imminent for resource-based cities to perform a high-quality transformation. The analysis of carbon emission efficiency in the Yellow River Basin includes the examination of spatiotemporal evolution characteristics and the main driving factors. This is done by utilizing the super-efficiency SBM-DEA and panel Tobit regression models, with the assistance of night light data. Our findings are as follows: (1) Carbon emissions continue to grow. The "Jiziwan" basin is an area where plenty of high-emitting cities agglomerate. The carbon emission of resource-based cities presents a W-shaped pattern in time. (2) In time, the carbon emission efficiency follows a U-shaped curve. Spatially, the carbon emission efficiency in the middle reaches is comparatively low, whereas it is relatively high in both the upper and lower reaches. And that in high carbon-emitting resource-based cities are in the low to medium range. (3) Carbon emission efficiency has a significant negative relationship with energy intensity, urbanization rate, and population density and a significant positive relationship with industrial proportion. Energy intensity is the most direct driving force. That is to say, we can increase carbon emission efficiency effectively by reducing energy intensity. [ABSTRACT FROM AUTHOR]

Objectives: Advanced cancer patients face various symptoms, which can cause physical and psychological distress. As a multidimensional construct, spiritual well-being (SWB) may be an inner resource for dealing with these problems. Our study explored the impact of different dimensions of SWB on physical and psychological symptoms in advanced cancer patients admitted to a palliative care unit. Methods: This cross-sectional study was conducted among 108 advanced cancer patients in the Hospice Ward, Shengjing Hospital of China Medical University. Patients completed questionnaires on SWB and cancer-related symptoms (insomnia, fatigue, pain, depression and anxiety) at the time of admission. Linear regression analysis was applied to determine the relationship between SWB (meaning, peace and faith) and symptom distress. Results: SWB accounted for an additional variance of cancer-related symptoms (17.8% to 44.4%). Meaning was negatively associated with insomnia (β = -0.516, p < 0.001) and fatigue (β = -0.563, p < 0.001). Peace and faith were related to lower psychological symptoms, while meaning represented a positive effect on anxiety (β = 0.275, p = 0.036). Higher peace was associated with lower cancer pain (β = -0.422, p < 0.001). Conclusions: Our findings suggested that achieving peace and faith appeared to function consistently as a positive resource for advanced cancer patients on depression, anxiety and pain, while meaning may serve to facilitate or hinder positive adjustment. Future studies should focus on the potential clinical implications by identifying the distinct dimension of SWB as symptom management targets in the palliative care practice. [ABSTRACT FROM AUTHOR]

Tembusu virus (TMUV) is an emerging flavivirus that has broken out in different regions of China. TMUV infection has been reported to induce autophagy in duck embryo fibroblast cells. However, the molecular mechanisms underlying this autophagy induction remain unclear. Here, we explored the interactions between autophagy and TMUV and the effects of the structural and nonstructural proteins of TMUV on autophagy in vitro. Among our results, TMUV infection enhanced autophagy to facilitate viral replication in HEK293T cells. After pharmacologically inducing autophagy with rapamycin (Rapa), the replication of TMUV increased by a maximum of 14-fold compared with the control group. To determine which TMUV protein primarily induced autophagy, cells were transfected with two structural proteins and seven nonstructural proteins of TMUV. Western blotting showed that nonstructural proteins 2B (NS2B) and 4 A (NS4A) of TMUV significantly induced the conversion of microtubule-associated protein 1 light chain 3 (LC3) from LC3-I to LC3-II in HEK293T cells. In addition, through immunofluorescence assays, we found that NS2B and NS4A significantly increased the punctate fluorescence of GFP-LC3-II. Furthermore, we found that both NS2B and NS4A interacted with polyubiquitin-binding protein sequestosome 1 (SQSTM1/p62) in a coimmunoprecipitation assay. Moreover, the autophagic degradation of p62 and LC3 mediated by NS2B or NS4A was inhibited by treatment with the autophagic flux inhibitor chloroquine (CQ). These results confirmed the vital effects of NS2B and NS4A in TMUV-induced complete autophagy and clarified the importance of complete autophagy for viral replication, providing novel insight into the relationship between TMUV and autophagy. [ABSTRACT FROM AUTHOR]

Background/objective: This multicenter study aimed to explore the efficacy and toxicity of radioactive Iodine-125 seed implantation for lymph node recurrence in patients with esophageal cancer after external radiotherapy. Methods: Clinical data of eligible patients from 5 centers in China were retrospectively reviewed. A total of 126 patients between January 2016 and March 2019 were included. The median interval between previous radiotherapy and radioactive Iodine-125 seed implantation was calculated. The target volume was 2.1–128.1 cm3 (median, 22.2 cm3) and the median postoperative D90 is 120.6 Gy (range, 101.7–192). Short-term efficacy of tumor response, the long-term efficacy of local progression-free survival (LRFS) and overall survival (OS), and treatment-related toxicity were reported. Results: For tumor response, 37 (29.4%), 51 (40.5%), 14 (11.1%), and 24 (19.0%) patients achieved complete response, partial response, stable disease and progressive disease, respectively. The 1-, 2- and 3-year LPFS and OS rates were 48.8%, 23.0% and 15.9%, and 80.2%, 38.8%, and 24.5%, respectively. Multivariate analysis identified Karnofsky performance status (P = 0.041) and tumor response (P = 0.049) as independent prognostic factors for LPFS; initial tumor stage (P = 0.034), lesion volume (P = 0.017), and tumor response (P = 0.004) as independent prognostic factors for OS. In total, 77 (61.1%) patients suffered from skin reactions and the incidence of grade 3–5 skin toxicity was 5.6% (7/126). Conclusion: Radioactive Iodine-125 seed implantation seems efficient with acceptable toxicity for the treatment of lymph node recurrence secondary to esophageal cancer. A head-to-head study is needed to further evaluate the survival benefit. [ABSTRACT FROM AUTHOR]

Increasing attention to serious environmental problems has caused the public to put forth more pressure on firms to fulfill their responsibilities to society at large. The establishment of a strict and mandatory environmental information disclosure (EID) mechanism has become particularly urgent. This study takes A-share listed firms in China's pharmaceutical manufacturing industry from 2011 to 2019 as a case. Considering the impact of the quality of EID on financial performance (FP), the moderating effects of internal and external factors, such as environmental regulation (ER), media pressure (MP), and executive compensation (EC), are analyzed. The results showed that the quality of EID had a negative effect on FP. It is difficult for EC to have a moderating effect on the relationship between the two. ER and MP, which are intended to exert strong external means of governance, cannot have a positive impact. Differences in regional marketization and ownership affect the moderators. This study highlights the difficulty of improving the environmental behavior of firms even while under the direct pressure from internal and external stakeholders and provides a reference for regulatory authorities to improve the EID mechanism. [ABSTRACT FROM AUTHOR]

Duck plague virus (DPV), a member of the alphaherpesvirus subfamily, can cause severe damage and immunosuppression in ducks and geese in China. Since lacking an available cell model, the antiviral signal transduction pathways induction and regulation mechanisms related to DPV infection in duck cells are still enigmatic. Our previous study developed a monocyte/macrophages cell model, which has been applied to study innate immunity with DPV. In the present study, we compared and analyzed transcriptome associated with the DPV infection of CHv (virulent strain) and CHa (avirulent strain) at 48hpi based on the duck monocyte/macrophages cell model and RNA-seq technology. Differentially expressed genes (DEGs) analysis showed 2,909 and 2,438 genes altered in CHv and CHa infected cells compared with control cells. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the DEGs were mainly involved in biological processes such as metabolic pathways, viral infectious diseases, immune system, and signal transduction. The CHv and CHa virus differentially regulated MAPK, NF-κB, and IFN signaling pathways based on transcriptome sequencing data and RT-qPCR results. The JNK inhibitor SP600125 enhanced the IFN signaling, but potentially reduced the VSV and DPV titers in the cell culture supernatant, indicating that JNK negatively regulates the IFN pathway and the inflammatory pathway to promote virus proliferation. The research results may provide promising information to understand the pathogenesis of DPV and provide a novel mechanism by which DPV modulates antiviral signaling and facilitate virus proliferation through hijacking the JNK pathway, which provides a new means for the prevention and control of DPV infection. [ABSTRACT FROM AUTHOR]

Purpose: Although spiritual well-being (SWB) is gaining increasing attention within the international palliative care (PC) guidelines, a lack of insight exists into the correlates and course of SWB among cancer patients. We therefore conducted a prospective observational study to capture trend of SWB and to identify their predictors in Chinese inpatients with terminal cancer receiving short-term PC. Methods: A prospective observational study was conducted of terminal cancer inpatients in the hospice ward, Shengjing Hospital of China Medical University. A total of 108 patients completed self-report questionnaires on Functional Assessment of Chronic Illness Therapy-Spiritual Well-being, Hospital Anxiety and Depression Scale, Numerical Rating Scales, and Life Orientation Scale-Revised anonymously at baseline; SWB, depression, anxiety, and pain were subsequently assessed at 1-week interval. Multilevel regression was used to analyze the temporal course and predictors of SWB. Results: Patients' existential well-being (B = − 0.99, p = 0.008; 95%CI = − 1.72 to − 0.26) and meaning dimension (B = − 0.87, p < 0.001; 95% CI = − 1.29 to − 0.43) significantly decreased after admission to the PC unit, but peace and faith did not change over time. Increases in depression and pain were related to lower existential well-being, particularly in the meaning dimension. Optimism–pessimism moderated the linear trend of existential well-being and meaning domain, such that those with higher optimism and lower pessimism paired with a decrease in outcomes. Conclusions: Terminal cancer patients experienced worsening existential well-being, particularly in the meaning facet while hospitalized, indicating that PC should include content that targets the existential concerns of spirituality in China. These findings also supported the need for an integrated PC to address personality traits and emotional and physical distress in this population. [ABSTRACT FROM AUTHOR]

Nowadays, firms spend a great deal of effort on Corporate Environmental Responsibility (CER) disclosure. From prospect theory, firms might disclose more CER information when they fail to achieve expected financial performance of the organization. We have constructed a CER disclosure index based on the "Guidelines for Environmental Information Disclosure of Listed Companies" issued by the Ministry of Environmental Protection of China in 2010. Using content analysis, we analyze 397 Chinese manufacturing Shanghai A-share listed firms from 2011 to 2017 (2090 firm-year observations) to investigate the relationship between corporate expected financial performance and CER disclosure. Our results show that, firstly, when the firm does not achieve the expected performance that is when the actual expectation is lower than the expected performance of the organization, the disclosure of corporate environmental responsibility increases; secondly, we have explored the moderating effect of industry categories; we have found that when firms fail to achieve the expected performance of the organization, light polluting firms are more inclined towards disclosing the environmental responsibility information than heavy polluting firms. Additionally, we have also studied the behavior of corporate environmental responsibility disclosure in the case of analysts expecting performance as a point of reference and got the same results. [ABSTRACT FROM AUTHOR]

Mountainous areas have become tourism hotspots in recent years, at the same time, some tourism safety issues have been revealed. To study rainstorm hazard prevention and mitigation capabilities, four representative scenic areas in the Qinling Mountains of China were selected as research foci for this study. The similar weight method and the unascertained measure model were used to establish an evaluation index, and the weights of indices at all levels were determined. Rainstorm hazard prevention and mitigation capabilities in the four scenic areas and on defferent slopes were then evaluated quantitatively. The results found that : according to the rainstorm hazard prevention and mitigation capabilities, the rankings of these scenic areas from high to low were Taibai Mountain National Forest Park, Cuihua Mountain National Geopark, Huayang Scenic Area, and Jinsixia National Forest Park. The rainstorm hazard prevention and mitigation capability on the north of the Qinling Mountains were higher than on the south slope, and the difference was relatively large. Rainstorm hazard prevention and mitigation capabilities on the eastern and western Qinling Mountains were more similar. These conclusions could provide a reference for regional rainstorm hazard prevention and mitigation measures, as well as sustainable development. [ABSTRACT FROM AUTHOR]

Harmonious ecological environment is a major concern with rising feeding and consumption of ducks, as these waterfowl birds can promote the spread of antibiotic resistant genes (ARGs). Therefore, this study was conducted to know diversity of antimicrobial resistance (AMR), integrons, and mobile genetic elements (MGEs) in Escherichia coli (E. coli) isolated from intestinal contents or pericardial effusion of diseased ducks from 2018 to 2020 in Sichuan, China. The AMR phenotype was determined via disk diffusion test in 165 E. coli isolates. Further, the integrase genes of integron (intI1 , intI2 and intI3 genes), gene cassettes (GCs) and MGEs were screened by PCR and sequencing. The results indicated 100% isolates were resistant to at least one antibiotic and 98.8% were multidrug-resistant strains. Highest AMR phenotype was recorded to rifampin (97.0%) followed by ampicillin (95.8%), chloramphenicol (89.7%), trimethoprim-sulfamethoxazole (84.2%), ciprofloxacin (83.0%), cefotaxime (80.0%), streptomycin (75.8%), doxycycline (49.7%), amikacin (10.3%), amoxicillin/clavulanic acid (3.6%), polymyxin B (1.2%) and ertapenem (0.6%). Further, class 1 and 2 integrons were found in 87.3% and 17.6% isolates, respectively. All isolates were negative for intI3 gene. The variable region of class 1 and 2 integrons contained total 13 different GCs, including arr-3 + dfrA27 , dfrA1 + aadA1 , dfrA17 + aadA5 , dfrA12 , dfrA1 + sat2 + aadA1 , dfrA12 + aadA2 , dfrA5 , aadA2 + ere(A) + dfrA32 , aac(6') - Ib - cr , aadA22 , aadA5 , dfrA17 , and dfrA27. Moreover, 13 MGEs in 69 different combinations were observed with predominance of IS 26 followed by tnpA /Tn 21 , trbC , IS Ecp1 , merA , IS Aba1 , tnsA , tnsB , tnsC , IS 1133 , tnsD , IS CR3 / 14 , and tnsE. Thus, the monitoring of integrons, MGEs and ARGs is important to understand the complex mechanism of AMR, which might help to introduce interventions for prevention and control of AMR in duck farms in China. [Display omitted] • 13 MGEs in 69 different combinations were observed from E. coli of diseased ducks. • Resistance phenotypes to SXT and C were found associated with integrons existence. • High diversity of integrons-GCs in MDR E. coli isolates were recorded. • The spreading of MGEs in the commensal intestinal bacteria was predictable. • Interventions are urgently required to limit antimicrobial use in duck industry. [ABSTRACT FROM AUTHOR]

There are rising concerns about microbes harboring antibiotic resistance genes (ARGs) and virulence-associated genes (VAGs) in humans and food-producing animals. Moreover, ARGs are considered as emerging environmental pollutants, posing probable life-threatening complications in humans and animals. Commensal Escherichia coli (E. coli) strain can carry a large number of VAGs, which may become opportunistic pathogen. The objective of this study was to determine the prevalence and possible association of ARGs and VAGs in E. coli isolates from clinically healthy waterfowls in China's tropical island, Hainan. For this purpose, 311 non-repeating E. coli isolates were evaluated for phenotypic drug resistance linked with ARGs. Additionally, strains were examined for subsequent resistance and virulence genes by uniplex or multiplex PCR and sequencing. Overall, 89 types of antibiotic resistance patterns were analysed, while 25 ARGs and 23 VAGs were observed, of which qnrS (99.4%) and iucD (99.7%) were the most commonly found genes, respectively. Significant positive associations were observed among ARGs and VAGs (p< 0.05, OR>1). The strongest association between resistance and virulence gene was observed for qnrS and iss (OR, 76.25; 95% CI, 4.02–1445.42). Our results propose that waterfowls serve as a reservoir of E. coli carrying multi ARGs and various ExPEC associated VAGs. Therefore, this study provides necessary information on the occurrence and possible associations of ARGs and VAGs in healthy waterfowls, which may act as a reference for the regulatory use of antibiotics to stop the direct or indirect spread of these resistant and potential virulent microbes to natural environment. • Prevalence of various ARGs and VAGs was studied in waterfowls. • A range of significant associations among ARGs and VAGs were detected. • 89 types of different antibiotic resistance patterns were observed. • The healthy waterfowl birds are possible reservoirs of VAGs. [ABSTRACT FROM AUTHOR]

The article focuses on Tongji University School of Medicine has established affiliation relationships with community health centers (CHCs) in Shanghai, which was unprecedented in China. Topics include the CHCs in strengthening their portfolio through the improvement of public trust, working environment, health care process and quality, professional development, clinical rotations, and community-oriented research, and CHCs are the cornerstones of the primary care system in urban China.

Duck Tembusu virus (DTMUV) has recently appeared in ducks in China and the key cellular determiners for DTMUV replication in host cells remain unknown. Autophagy is an evolutionarily conserved cellular process that has been reported to facilitate flavivirus replication. In this study, we utilized primary duck embryo fibroblast (DEF) as the cell model and found that DTMUV infection triggered LC3-II increase and polyubiquitin-binding protein sequestosome 1 (p62) decrease, confirming that complete autophagy occurred in DEF cells. The induction of autophagy by pharmacological treatment increased DTMUV replication in DEF cells, whereas the inhibition of autophagy with pharmacological treatments or RNA interference decreased DTMUV replication. Inhibiting autophagy enhanced the activation of the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and interferon regulatory factor 7 (IRF7) pathways and increased the p62 protein level in DTMUV-infected cells. We further found that the overexpression of p62 decreased DTMUV replication and inhibited the activation of the NF-κB and IRF7 pathways, and changes in the NF-κB and IRF7 pathways were consistent with the level of phosphorylated TANK-binding kinase 1 (p-TBK1). Opposite results were found in p62 knockdown cells. In summary, we found that autophagy-mediated p62 degradation acted as a new strategy for DTMUV to evade host innate immunity. [ABSTRACT FROM AUTHOR]

Duck tembusu virus (DTMUV) is a single-stranded, positive-polarity RNA flavivirus that has caused considerable economic losses in China in recent years. Innate immunity represents the first line of defense against invading pathogens and serves as an important role in resisting viral infections. In this study, we found that the infection of ducks by DTMUV triggers Toll-like receptors (TLRs) and (RIG-I)-like receptors (RLRs) signaling pathways and inducing abundant of pro-inflammatory factors and type I interferons (IFNs), in which melanoma differentiation-associated gene 5 (MDA5) and Toll-like receptor 3 (TLR3) play important immunity roles, they can inhibit the replication process of DTMUV via inducing type I IFNs. Moreover, we demonstrated that type I IFNs can inhibit the DTMUV replication process in a time- and dose-dependent manner. Exosomes are small membrane vesicles that have important roles in intercellular communication. MicroRNAs (miRNAs) are small non-coding RNAs that can modulate gene expression and are common substances in exosomes. In our experiment, we successfully isolated DEF cells derived exosome for the first time and explored its function. Firstly, we found the expression of miR-148a-5p is significantly decreased following DTMUV infect. Then we found miR-148a-5p can target TLR3 and down-regulate the expression of TLR3, serving as a negative factor in innate immunity. Unfortunately, we cannot find miRNAs with different expression changes that can target MDA5. Lastly, our experimental results showed that TLR3 was one of the causes of miR-148a-5p reduction, suggesting that the high level of TLR3 after DTMUV infect can both trigger innate immunity and suppress miR-148a-5p to resist DTMUV. [ABSTRACT FROM AUTHOR]

Background: Riemerella anatipestifer is one of the most serious infectious disease-causing pathogens in the duck industry. Drug administration is an important method for prevention and treatment of infection in duck production, leading to widespread drug resistance in R. anatipestifer. Methods: For a total of 162 isolates of R. anatipestifer, the MICs were determined for a quinolone antimicrobial agent, namely, nalidixic acid, and three fluoroquinolones, namely, ciprofloxacin, enrofloxacin and ofloxacin. The gyrA, parC, and parE gene fragments were amplified by PCR to identify the mutation sites in these strains. Site-directed mutants with mutations that were detected at a high frequency in vivo were constructed (hereafter referred to as site-directed in vivo mutants), and the MICs of these four drugs for these strains were determined. Results: In total, 100, 97.8, 99.3 and 97.8% of the 137 R. anatipestifer strains isolated between 2013 and 2018 showed resistance to nalidixic acid, ciprofloxacin, enrofloxacin, and ofloxacin, respectively. The high-frequency mutation sites were detected in a total of 162 R. anatipestifer strains, such as Ser83Ile and Ser83Arg, which are two types of substitution mutations of amino acid 83 in GyrA; Val799Ala and Ile811Val in ParC; and Val357Ile, His358Tyr, and Arg541Lys in ParE. MIC analysis results for the site-directed in vivo mutants showed that the strains with only the Ser83Ile mutation in GyrA exhibited an 8–16-fold increase in MIC values, and all mutants showed resistance to ampicillin and ceftiofur. Conclusions: The resistance of R. anatipestifer to quinolone agents is a serious problem. Amino acid 83 in GyrA is the major target mutation site for the fluoroquinolone resistance mechanism of R. anatipestifer. [ABSTRACT FROM AUTHOR]

Duck hepatitis A virus (DHAV) is prevalent worldwide and has caused significant economic losses. As the predominant serotype in China, DHAV-3 has become a major challenge to the local duck industry. Here the genetics and pathogenesis of a virulent DHAV-3 strain and its embryo-passaged strain were assessed. There were only two amino acid substitutions (Y164N in VP0 protein and L71I in 2C protein) introduced during the adaptation process. The pathogenicity of these strains was further evaluated in vivo. Clinical signs, gross pathology, and histopathological analysis showed that the embryo-passaged strain was attenuated. Meanwhile, the viral RNA loads were significantly lower in the liver tissues of the ducklings infected with the attenuated strain. As expected, infection with the virulent and attenuated strains led to the activation of different innate immune genes. We suspected that the loss of replication efficiency in ducklings was responsible for the attenuation phenotype of the embryo-passaged strain. In addition, different innate immune responses in the liver of ducklings were at least partly responsible for the differential infectivity phenotype. These findings provide new insights into the genetics and pathogenesis of DHAV-3, which may aid the development of new vaccines and the implementation of immunization strategies. [ABSTRACT FROM AUTHOR]

Background: Short beak and dwarfism syndrome (SBDS) was caused by novel goose parvovirus (NGPV)--a variant of goose parvovirus (GPV). Ducks infected with NGPV shows clinical signs including growth retardation and protrusion of the tongue from an atrophied beak. SBDS outbreak was first reported at the northern coastal provinces of China during 2015 and it was again reported in Sichuan, an inland province of China in 2016. The disease caused a huge economic loss in Chinese duck feeding industry. Results: The SD15 strain of NGPV was isolated from liver and intestinal tract tissue samples of infected ducks. Real-time quantitative PCR (qPCR) was used to estimate viral load in embryonated eggs and cells infected with adapted virus. The data showed that duck embryo fibroblasts (DEFs) were permissive to NGPV, while goose embryo fibroblasts (GEFs) cells were not, and the copy numbers of SD15 in the allantoic fluid of infected eggs remained at 105.0–106.5 copies/ml. The adaption procession of the virus was determined via qPCR, and viral proliferation was detected through indirect fluorescent antibody assay (IFA) in DEFs. It was further determined that viral copy numbers peaked at 96 h post-inoculation (hpi), which is the best time to harvest the virus in DEFs. Cytotoxic effects and cell death were observed at 72 hpi in SD15 infected DEFs, yet SD15 did not induce apoptosis. Conclusions: The growth characteristics of SD15 strain of NGPV determined would be beneficial for further molecular characterization of these viruses and develop potential vaccines if required. [ABSTRACT FROM AUTHOR]

Goose astrovirus (GAstV) is a newly identified viral pathogen threatening waterfowl, exhibiting a high prevalence across various regions in China. Notably, the Guanghan District of Deyang City, situated in Sichuan Province, has faced a outbreak of GAstV, resulting in significant mortality among goslings due to the induction of gout-like symptoms. In our research, we successfully isolated a GAstV strain known as GAstV SCG3. This strain exhibits efficient replication capabilities , proving virulent in goslings and goose embryos. Our study delved into the characteristics of GAstV SCG3 both in vitro and in vivo. Additionally, we examined tissue phagocytosis and the distribution of GAstV SCG3 in deceased goslings using H&E staining and IHC techniques. According to the classification established by the ICTV, GAstV SCG3 falls under the category of GAstV genotype-2. Notably, it demonstrates the highest homology with the published AHAU5 sequences, reaching an impressive 98%. Furthermore, our findings revealed that GAstV SCG3 exhibits efficient proliferation exclusively in goose embryos and in LMH cells, while not manifesting in seven other types of avian and mammalian cells. Significantly, the mortality of GAstV on goslings and goose embryos are 93.1 and 80%, respectively. Moreover, the viral load in the livers of infected goslings surpasses that in the kidneys when compared with the attenuated strain GAstV SCG2. The mortality of GAstV is usually between 20% and 50%, our study marks the first report of a virulent GAstV strain with such a high mortality. [ABSTRACT FROM AUTHOR]

Highlights • First report on the prevalence and genetic characteristics of ESBL producing E. coli isolates from waterfowl birds in China. • In this study, 92.9% of the E. coli isolates were confirmed as ESBL phenotypes by double-disk synergy testing. • CTX-M, belonging to STs associated with E. coli infections should be of concern to animal and public health. Abstract The epidemiology and genetic characteristics of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli (E. coli) have been widely studied in human and veterinary settings. ESBL-producing E. coli are generally reported in pigs, poultry, and dairy farm animals. Here, we report on the prevalence and genetic characteristics of beta-lactamase producing E. coli isolated from waterfowl birds in Hainan, China. After phenotypic confirmation, genes encoding bla CTX- M , bla SHV and bla TEM were detected by polymerase chain reaction (PCR). The isolates were assigned to different phylogenetic groups, and multi-locus sequence typing (MLST). Taken as a whole, 289 (92.9%) out of 311 E. coli isolates from waterfowl birds were confirmed as ESBL phenotypes by double-disk synergy testing. Subsequent PCR analysis revealed that bla CTX- M was the predominant ESBL gene identified in 146 (46.9%) isolates, followed by the combination of bla CTX- M and bla TEM in 70 (22.5%) isolates. The majority of these positive isolates were assigned to phylogroup B2 (46.2%) followed by phylogroup A (43.6%). In addition, MLST assigned representative ESBL positive isolates (n = 40) to 18 STs, and ST410 (ST23cplx) was the most prevalent population (22.5%). The high prevalence of CTX-M and STs frequently associating with E. coli infections should be of concern as it poses threats to animal and public health. To the best of our knowledge, this is the first comprehensive study to report on the occurrence of ESBL producing E. coli from waterfowl birds in China. [ABSTRACT FROM AUTHOR]

Identification and analysis of the avian reovirus from black swan. Isolation of the strain through the chorioallantoic membrane route of duck embryos, identified through transmission electron microscopy and RT-PCR based on the ARV S2 gene. The complete genome of the ARV strain was obtained using next-generation sequencing technology. The isolated strain of ARV was named CD200801 and was identified through transmission electron microscopy and RT-PCR based on the ARV S2 gene. Experimental infection with CD200801 resulted in the death of ducklings with serious spleen and liver focal necrosis. BLAST analysis of CD200801 sequences showed a 35.5 to 98.6% similarity to a novel duck reovirus that was isolated in recent years. Phylogenetic analysis revealed that CD200801 was closely related to ARV isolates YL, GX-Y7, and XT-18. We report the first avian reovirus infection in the black swan. This study provides important new insights into the evolutionary relationships among different ARV strains and highlights the need for continued surveillance and monitoring of these viruses in both domestic and wild bird flocks. These findings have significant implications for the development of effective strategies for disease prevention and control in the poultry industry. [ABSTRACT FROM AUTHOR]

Since 2010, the duck Tembusu virus (DTMUV) has caused a severe outbreak of egg drop syndrome in laying ducks in China, which has resulted in substantial financial losses in the poultry industry. DTMUV nonstructural protein 1 (NS1), as the only secreted protein, could aid in the development of therapeutic antibodies and diagnostic techniques; however, there are few studies on the preparation and epitope identification of monoclonal antibodies (mAbs) against DTMUV NS1. In this study, by indirect enzyme-linked immunosorbent assay (ELISA), Western blotting, and indirect immunofluorescence assay, we screened 6 mAbs (8A4, 8E6, 10F12, 1H11, 3D5, 5C11) that could specifically recognize DTMUV NS1. For epitope mapping of mAbs, a series of GST-tagged truncated fusion proteins of DTMUV NS1 were constructed by prokaryotic expression. Finally, the 4 shortest linear epitopes were identified by indirect ELISA and Western blotting. The epitope 133FVIDGPK139 was recognized by 8A4, the epitope 243IPKTLGGP250 was recognized by 8E6, the epitope 267PWDEK271 was recognized by 10F12, and 156EDFGFGVL163 was recognized by 1H11, 3D5, and 5C11. By sequence alignment and cross-reaction tests, we found that 8A4 and 8E6 had high specificity for DTMUV NS1 compared with that of other mAbs, but 10F12, 1H11, 3D5, and 5C11 exhibited a clear degree of cross-reaction with dengue virus (DENV), Japanese encephalitis virus (JEV), West Nile virus (WNV), and Zika virus (ZIKV) NS1. Finally, the predicted crystal structure analysis showed the approximate spatial positions of the 4 epitopes on the NS1 dimer. In summary, our study revealed 2 specific mAbs for DTMUV NS1 recognition and 4 multiflavivirus mAbs for DENV, JEV, WNV, and ZIKV NS1 recognition. [ABSTRACT FROM AUTHOR]

Duck Tembusu virus (DTMUV), belonging to the Flaviviridae family, is a major virus that affects duck health in China. MicroRNAs (miRNAs) play an important role in viral replication. However, little is known about the function of miRNAs during DTMUV infection. Here, the host miR-146b-5p was found to regulate DTMUV replication. When DTMUV infected duck embryo fibroblasts (DEFs), the expression levels of miR-146b-5p increased significantly over time. Moreover, the viral RNA copies, E protein expression levels and virus titers were all upregulated when miR-146b-5p was overexpressed in DEFs. The opposite results were also observed upon knockdown of miR-146b-5p in DEFs. To explore the mechanism by which miR-146b-5p promoted DTMUV replication, mass spectrometry, and RNA pull-down assays were employed. Ribosomal protein S14 (RPS14), a component of 40S ribosomal proteins, was identified to interact with miR-146b-5p. In addition, the relative mRNA expression levels of RPS14 gene were negatively modulated by miR-146b-5p. Subsequently, it was found that overexpression of RPS14 could decrease the replication of DTMUV, and the reverse results were also detected by knockdown of RPS14. In conclusion, this study revealed that miR-146b-5p promoted DTMUV replication by targeting RPS14, which provides a new mechanism by which DTMUV evades host defenses and a new direction for further antiviral strategies development. [ABSTRACT FROM AUTHOR]

Duck plague is an acute, febrile, and septic infectious disease caused by duck plague virus (DPV), which causes serious harm to the duck industry in China. Ducks latently infected with DPV display a clinically healthy state, which is one of the epidemiological characteristics of duck plague. In the present study, to rapidly distinguish vaccine-immunized ducks from wild virus-infected ducks during production, a PCR assay based on the newly identified LORF5 fragment was developed to effectively and accurately identify viral DNA in cotton swab samples and was used to assess artificial infection models and clinical samples. The results showed that the established PCR method had good specificity and that only the virulent and attenuated DNA of duck plague virus was specifically amplified, as the results for the detection of common duck pathogens (duck hepatitis B virus, duck Tembusu virus, duck hepatitis A virus type 1, novel duck reovirus, Riemerella anatipestifer, Pasteurella multocida , and Salmonella) were negative. The amplified fragments of virulent and attenuated strains were 2,454 bp and 525 bp, and their minimum detection amounts were 0.46 pg and 46 pg, respectively. The detection rate of the virulent and attenuated DPV strains in duck oral and cloacal swabs was lower than that of the gold standard PCR method (GB-PCR , which is unable to distinguish virulent and attenuated strains), and cloacal swabs from clinically healthy ducks were more suitable for detection than oral swabs. In conclusion, the PCR assay established in the present study can be used as a simple and effective method for the clinical screening of ducks that are latently infected with virulent strains of DPV and shedding virus, which can provide technical support for the elimination of duck plague from duck farms. [ABSTRACT FROM AUTHOR]

Identification and analysis of the antimicrobial resistance of Pasteurella aerogenes (P. aerogenes) isolated from poultry. For susceptibility testing in accordance with the CLSI, plasmids were extracted via alkaline lysis and transferred by CaCl 2 treatment. Genomic DNA of a representative P. aerogenes isolate was subjected to whole genome sequencing. CCCP was utilized to determine whether SF190908 contains an efflux pump. The bla VEB gene was ligated with the pET-28 plasmid and transferred to Escherichia coli to verify it as an ESBL gene. SF190908 isolated from poultry was identified as P. aerogenes based upon biochemical and 16s rRNA results. The isolate showed high MIC values for eight antimicrobials. Sequencing results showed that the mobile element-mediated antimicrobial resistance gene cluster conferred antimicrobial resistance on the strain, and a single 5,105-bp plasmid, designated pRCAD0752PA-1, was isolated. Four antimicrobial resistance gene clusters were identified in the SF190908 chromosome; one antimicrobial resistance gene cluster carried the bla VEB gene, which was verified as ESBL according to the CLSI and was detected in Pasteurellaceae for the first time, to the best of our knowledge. The efflux pump may confer antimicrobial resistance to SF190908. P. aerogenes isolated from poultry showed resistance genes encoded in mobile elements that confer multi-antimicrobial resistance to SF190908. The antimicrobial-resistant plasmid pRCAD0752PA-1 was isolated in SF190908 and conferred resistance to florfenicol. This study indicates an urgent need to increase efforts to monitor the spread of P. aerogenes multi-antimicrobial-resistant strains and plasmids, especially in newly discovered at-risk species such as poultry. [ABSTRACT FROM AUTHOR]

Bacterial antimicrobial resistance (AMR) continues to develop, with the horizontal transfer of antibiotic resistance genes (ARG s) through plasmids playing a major role. Recently, the antimicrobial resistance of R. anatipestifer has become increasingly severe, jeopardizing the development of the poultry industry. In this study, we used PromethION to determine the whole genome sequence of R. anatipestifer RCAD0416, a multidrug-resistant isolate from China. We detected a plasmid in the isolate. We named the plasmid pRCAD0416RA-1; the plasmid was 37356 bp in size with 36 putative open reading frames and included the bla OXA-347 , flo R , tet (X) , erm F , ere D, and Aad S resistance genes. Most resistance genes might be obtained from R. anatipestifer HXb2. Mobile elements and floR might be transmitted by plasmid pB18–2 from Acinetobacter indicus , and the ICEPg6Chn1 mobile elements can be transmitted from Proteus genomosp. The plasmid pRCAD0416RA-1 was transferred to Escherichia coli K-12 × 7232 via electroporation. Subsequent antimicrobial sensitivity tests (AST) showed a noticeable levels of antimicrobial resistance to β-lactams (4–8 fold), tigecycline (8 fold), and florfenicol (8 fold). These types of antibiotics are in common clinical use. The purpose of this article is to elucidate the basic characteristics of pRCAD0416RA-1 and the level of resistance mediated by bla OXA-347 , flo R, and tet (X). [ABSTRACT FROM AUTHOR]

• First study on bacterial diversity of zha-chili using high-throughput sequencing. • Bacterial communities in zha-chili are rich and diverse. • The place of origin affects the bacterial community structure and flavor of zha-chilli. • Lactobacillus dominates the bacterial community, metabolism and flavor of zha-chilli. Zha-chili is a traditional Chinese fermented food with special flavor, which is often used as an appetizer in condiments and an important energy source. The final quality of zha-chili is closely related to its microbial community structure. However, the differences of bacterial diversity in zha-chili from different regions and how bacterial species affect zha-chili fermentation process and flavor quality have not been reported. In this study, the bacterial diversity and flavor quality of zha-chili samples from different regions were analyzed using Illumina Miseq high-throughput sequencing, electronic nose and electronic tongue technology. Twenty-three bacterial phyla and 665 bacterial genera were identified in all zha-chili samples. Firmicutes, Proteobacteria and Actinobacteria were the dominant bacterial phyla in zha-chili samples, while Lactobacillus , Pseudomonas , Pediococcus , Weissella and Staphylococcus were the dominant bacterial genera. The bacterial community structure of zha-chili samples from different regions was significantly diverse (p < 0.05). The flavor of zha-chili samples also varied in different regions, and the discrepancy of taste was much greater than that of aroma. Moreover, there were significant correlations (p < 0.05) between 6 dominant bacterial genera and 8 flavor indicators (3 aroma indicators, 5 taste indicators). In addition, the results of microbiome phenotypes prediction by BugBase and bacterial functional potential prediction using PICRUSt showed that eight out of nine predicted phenotypic functions of zha-chili samples from different regions were significantly different (p < 0.05), bacterial metabolism was vigorous in the zha-chili samples, and Lactobacillus was the dominant bacterial genus involved in metabolism during fermentation. [ABSTRACT FROM AUTHOR]

Duck Tembusu virus (DTMUV), an emerging infectious pathogen, has caused severe disease in ducks and huge economic losses to the poultry industry in China since 2009. Despite considerable advances in understanding the effects of microRNAs on host antiviral immune responses, it remains unclear how miRNAs regulate DTMUV replication in duck embryo fibroblast (DEF) cells. This study aims to clarify the role of host microRNA-148a-5p (miR-148a-5p) in regulating DTMUV replication by targeting SOCS1. First, we found that during DTMUV infection, the expression of miR-148a-5p in DEFs was downregulated in a time-dependent and dose-dependent manner, while the expression of SOCS1 was significantly upregulated. In addition, we found that when miR-148a-5p mimics were transfected into DEFs, viral RNA copies, viral E protein expression levels and viral titres, which represent viral replication and proliferation, were significantly downregulated, while the opposite result was observed when miR-148a-5p inhibitor was transfected into DEFs. Next, we found that SOCS1 was the target gene of miR-148a-5p through software analysis. Therefore, we further confirmed that SOCS1 was the target of miR-148a-5p and that miR-148a-5p could negatively regulate the expression of SOCS1 at the mRNA and protein levels. Furthermore, our results indicated that overexpression of SOCS1 promoted DTMUV replication, while knockdown of SOCS1 inhibited DTMUV replication. Finally, we found that in DTMUV-infected DEFs, the overexpression of SOCS1 inhibited the production of IFN-α and IFN-β, while knocking down SOCS1 produced the opposite result. This indicates that during DTMUV infection, the virus promotes the expression of SOCS1 by downregulating the expression of miR-148a-5p, while the upregulation of SOCS1 suppresses the production of type I interferon and promotes virus replication. Taken together, these findings provide new insights into virus-host interactions during DTMUV infection and provide potential new antiviral treatment strategies for DTMUV infection. • In this study, we first verified the expression kinetics of miR-148a-5p and SOCS1 in DEF cells infected with DTMUV. • Next, we explored the direct effect of miR-148a-5p on DTMUV replication for the first time. • Futhermore, we found that miR-148a-5p negatively regulated SOCS1 mRNA and protein expression levels. Finally, we found that SOCS1 promoted viral replication by inhibiting the production of type I interferon. [ABSTRACT FROM AUTHOR]

• Prevalence of NDM-1-producing E. coli in waterfowls. • The waterfowls are possible reservoirs of bla NDM-1. • Five different STs and 6 PFGE subtypes were observed. • PFGE patterns suggested the propagation of identical clone between the various farms. Carbapenems are traditionally recognized to be the last resort drugs to treat infections due to MDR organisms such as E. coli. As such, the emergence of New Delhi metallo-β-lactamase-producing E. coli strains have become a challenging threat to the public health. In this regard, we examined the molecular characteristics of carbapenem-resistant E. coli (CRE) isolated from waterfowls in China's tropical island, Hainan. A total of 311 single E. coli strains were obtained from 20 various farms of healthy ducks and geese in 2 districts of Hainan island. The CRE strains were initially identified via phenotypic resistance and modified Hodge test. PCR assay and subsequent nucleotide sequencing were used to detect different types of carbapenemase encoding genes (bla NDM , bla VIM , bla IMP , bla OXA and bla KPC). In addition, MLST and PFGE analyses were also performed. Among the 311 E. coli strains, 8 strains were detected to produce a single type of carbapenemase i.e. NDM-1 (2.6%). A total of 5 sequence types (STs) were observed, of which ST10 was the most prevalent accounting for 37.5% (3/8). Moreover, these 8 isolates yielded 6 different PFGE clusters but showed approximately related PFGE types, suggesting the propagation of similar clone between the farms. This is the first report on the identification of NDM-1-producing E. coli from waterfowls in Hainan island, China. Our results emphasize the need for better efforts to control the further spread of NDM-1-producing E. coli strains in this tropical island. [ABSTRACT FROM AUTHOR]

Tigecycline is considered one of the "last resort antibiotics" for treating complex infections caused by multidrug-resistant (MDR) bacteria, especially for combating clinical resistant strains that produce carbapenemases. However, the tet(X4) gene, which carried by different plasmids can mediate high levels of bacterial resistance to tigecycline, was first reported in 2019. Here, we report the emergence of the plasmid-mediated tet(X4) in avian environment of Sichuan Province. A total of 21 tet(X4)-positive Escherichia coli (E. coli) strains were isolated and identified from avian samples in selected regions, with an isolation rate of 1.6% (21/1,286), and all of them were MDR strains. Multilocus Sequence Typing (MLST) method was used to classify the 21 tet(X4)-positive E. coli into the ST206, ST761, ST155, ST1638, ST542, and ST767 types, which also belong to the 3 phylogenetic subgroups A, B1, and C. Tet(X4) is located on mobile plasmids that can be efficiently and stably propagated. The results of fitness cost experiments showed that tet(X4)-positive plasmids may incur some fitness cost to host bacteria, but different tet(X4)-positive plasmids bring about differential fitness costs. Whole-genome sequencing further confirmed the tet(X4) gene can be located on IncX1-type plasmids and the core genetic structures are ISVsa3-rdmc-tet(X4) or rdmc-tet(X4)-ISVsa3, the former is a 7 copies tandem repeat structure. In this study, we isolated and identified tet(X4)-positive E. coli from the avian origin in Sichuan, analyzed the mobility of the tet(X4) by conjugational transfer and S1-PFGE, and evaluated the biological characteristics of the tet(X4)-positive plasmid using the results of conjugational frequency, plasmid stability, and fitness costs. Finally, combined with the third-generation whole-genome sequencing analysis, the molecular transmission characteristics of the tet(X4) were preliminarily clarified, providing a scientific basis for guiding veterinary clinical use in this area, as well as risk assessment and prevention of the transfer and spread of tigecycline resistant strains or genes., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

With the increasing number of ducks being raised and consumed, it is crucial to monitor the presence of multidrug resistant (MDR) bacteria in duck farming. Waterfowl, such as ducks, can contribute to the rapid dissemination of antibiotic resistance genes (ARGs). The objective of this study was to investigate the antimicrobial resistance (AMR), ARGs, and mobile genetic elements (MGEs), such as IS26, tbrC, ISEcp1 in Escherichia coli(E. coli) isolated from the intestinal contents of diseased ducks between 2021 and 2022 in Sichuan, Chongqing and Anhui, China. The AMR phenotypes of 201 isolated E. coli strains were determined using the minimum inhibitory concentrations (MICs) method. Subsequently, polymerase chain reaction and sequencing techniques were employed to screen for integron-integrase genes (intI1, intI2, intI3 genes), gene cassettes (GCs), MGEs, and ARGs. The results demonstrated that 96.5% of the E. coli isolates were resistant to at least 1 antibiotic, with 88.1% of the strains displaying MDR phenotype. The highest AMR phenotype observed was for trimethoprim-sulfamethoxazole (88.1%). Furthermore, class 1 and class 2 integrons were detected in 68.2% and 3.0% of all the isolates, respectively, whereas no class 3 integrons were found. Ten types of GCs were identified in the variable regions of class 1 and class 2 integrons. Moreover, 10 MGEs were observed in 46 combinations, with IS26 exhibiting the highest detection rate (89.6%). Among the 22 types of ARGs, tetA (77.1%) was the most frequently detected. In the conjugational transfer experiment, transconjugants were found to carry specific ARGs and MGEs, with their MIC values were significantly higher than those of recipient E. coli J53, indicating their status as MDR bacteria. This study emphasizes the necessity of monitoring MGEs, ARGs, and integrons in duck farms. It provides valuable insights into the complex formation mechanisms of AMR and may aid in preventing and controlling the spread of MDR bacteria in waterfowl breeding farm., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

We aimed to examine the longitudinal association between frailty and cognitive impairment in the older Chinese population. This prospective cohort study used data from the Chinese Longitudinal Healthy Longevity Study 2011 wave. We calculated the follow-up duration as 3 years from the baseline year. Frailty was measured using the frailty index, and cognitive function was calculated by Mini-Mental State Examination Scale. Participants who were non-frailty and those with normal cognitive function were included in 2011 and followed up in 2014, respectively. Frailty was an independent risk factor for early-onset cognitive impairment. Age, hearing impairment, and a decreased ability to perform daily activities were the main risk factors for cognitive impairment, while affluent economic status was a protective factor. Cognitive impairment was not found to be an independent risk factor for frailty. We concluded that the frailty index is a significant predictor of cognitive impairment among community-dwelling older adults.

The expansion of government size will have dual effects on a country's green innovation. An appropriately sized government size increases marginal productivity and stimulates the development of green innovation by increasing government expenditure. On the contrary, an excessively sized government creates a huge administrative agency, which not only increases the tax burden but also damages social welfare by excessive intervention. Therefore, the effect of government size on green innovation is not linear. In order to prove this proposition, this study examines the impact of government size on green innovation in 166 countries between 1995 and 2018, using a two-way fixed effects model. The results reveal an inverted U-shaped relationship between government size and the level of green innovation, indicating that optimal government size may maximize a country's green innovation output. The results further suggest that this inverted U-shaped relationship is mainly influenced by environmental regulations and financial support. Finally, our heterogeneity analysis demonstrates that the inverted U-shaped relationship is more pronounced for countries with high organizational inertia and more R&D expenditure than for those with low organizational inertia and less R&D expenditure. This finding makes up for the research gap between government size and green innovation and provides a reference for countries to formulate the optimal government size to improve the level of green innovation.

Based on the ultrasonic imaging and endoscopic resection of the intelligent segmentation algorithm, this study is aimed at exploring whether nursing intervention can promote the good recovery of patients with colon polyps, hoping to find a new method for clinical treatment of the colon polyps. Patients with colon polyps were divided into an experimental group (fine nursing) and a control group (general nursing). The colonoscopy polyp ultrasound image was preprocessing to select the seed points and background points. The random walk decomposition algorithm was applied to calculate the probability of each marked point, and then, the marked image was outputted. The accuracy of the intelligent segmentation algorithm was 81%. The incidence of complications in the experimental group was 4.83%, which was lower than 16.66% in the control group, and the difference was statistically obvious ( P < 0.05). Perioperative refined nursing intervention for colon polyp patients undergoing endoscopic electrosurgical resection can decrease postoperative adverse reactions; reduce postoperative mucosal perforation, blood in the stool, abdominal pain, and small bleeding; lower the incidence of postoperative complications; and allow patients to recover quickly, enhancing the life comfort of patient., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2021 Na Ma et al.)

Previously, we isolated a novel strain of goose pegivirus (GPgV) that infects geese and shows high levels of lymphotropism. This novel pegivirus strain is phylogenetically distinct from previously known Pegivirus species, Pegivirus A-K, and qualifies as a candidate new Pegivirus species, GPgV. GPgV is tentatively named Pegivirus M. Here, to better understand the epidemic of GPgV infection and the coinfection of this virus with other viruses in Southwest China, 25 geese in poor health from Sichuan Province and 24 geese in poor health from the municipality of Chongqing were collected. The geese were tested for 9 types of goose viruses (goose hemorrhagic polyomavirus, GPgV, astrovirus, parvovirus, circovirus, reovirus, coronavirus, paramyxovirus, and avian influenza virus) by RT-PCR or nested RT-PCR. GPgV RNA was detected in 2 out of 25 monoinfections and 8 out of 25 coinfections with other viruses on Sichuan farms and 2 out of 24 monoinfections and 10 out of 24 coinfections on Chongqing farms. Overall, 22 of the 49 (44.9%) geese were positive for GPgV, which indicated a high infection rate. To the best of our knowledge, this is the first report of GPgV coinfection with other epidemic viruses. This study enhances our understanding of the emergence and epidemiology of Pegivirus., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Nitrogen (N) deposition and the input of litter to soils are expected to significantly affect soil respiration (R s ). However, the responses of R s to N deposition and litter input alteration in semiarid grassland are still unclear. We investigated the effects of N addition, litter input alteration, and their interactions on R s throughout the growing season from May to September in 2017, in Horqin grassland, Northern China. N deposition included N addition (10 g N m - 2  year - 1 ) and N control (0 g N m - 2  year - 1 ), and litter alteration consisting of litter removal, litter doubling and litter control were conducted. We found that N addition significantly improved vegetation coverage (P < 0.01) and thus increased root respiration (R t ) by 30% (P < 0.05), but did not significantly enhance R s (P > 0.05) due to the negative effect of microbial respiration (R m ). Litter removal significantly decreased soil microbial biomass and thus reduced R m (13%) and R s (14%), but litter doubling did not significantly alter R m , R t and R s . N addition slightly weakened the inhibition of litter removal and led to soil respiration was not significantly decreased, whereas N addition and litter doubling did not significantly interact with R s . Furthermore, R s and its components were mainly mediated by soil temperature, moisture, soil microbial biomass carbon (SMBC) and vegetation coverage, but the structural equation model found that soil temperature, SMBC and coverage were the key controlling factor of R s , R m and R t , respectively. Different influences of N deposition and litter input on R s , as well as the contributions of R m and R t to R s identified could facilitate the evaluation and prediction of R s and belowground C storage in grassland ecosystems in the context of future global changes., Competing Interests: Declaration of competing interest We declare that we have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Transgenic maize expressing the Cry1Ab and Cry2Ab protein simultaneously from Bacillus thuringiensis (Bt-maize) has been grown for farm-scale study to investigate its potential impact to non-target arthropod (NTA). The trials were conducted between Bt maize 2A-7 and its parental line (B73-329) in Beijing, China, over 3 years. Richness (C), Shannon index (H), Pielou index (J), Simpson index (D), and Bray-Curtis index were used to evaluate the population dynamics and biodiversity of the dominant arthropods from per 50 plants in crop field. The mainly abundant groups were Aphidoidea, Araneae, Coccinellidae, Anthocoridae, and Thripidae which represented about 90% of the total number of NTA. Although the abundance of NTA varied from year to year, there is no significant difference between Bt maize and non-Bt maize field. Fluctuations were found at individual sample dates, but the trend of these descriptors remained consistent. Further analysis showed the biodiversity indexes of the dominant arthropods C, H, J, D, and Bray-Curtis dissimilarity between Bt maize producing Cry1Ab and Cry2Ab toxin simultaneously and its parental line had no significant difference except for some sampling dates. These results suggested that Bt maize is compatible with the NTAs and provides further evidence of the ecological impact of genetically modified maize.

Rice wine, a traditional fermented alcoholic beverage in China, is produced with grains such as rice, which are fermented with saccharifying starter-koji. Its flavor quality is closely associated to the starter culture-koji, which is made by mixing botanical materials with high-class glutinous rice in certain ecological context. However, there are few reports on the microbial community structure of rice wine koji. In this paper, bacterial community structures of rice wine koji were analyzed using 16S rRNA gene sequencing based on Illumina MiSeq high-throughput technology in 20 samples collected from Xiaogan area, Hubei province and Dazhu area, Sichuan province (10 from each area). We found rice wine koji flora mainly consisted of Weissella, Lactobacillus, Lactococcus, Bacillus, Enterococcus, and Cronobacter, with relative abundances of 29.49%, 10.93%, 8.85%, 4.75%, 1.16% and 1.15%, respectively, as well as an accumulative average relative abundance of 58.71%. They all belonged to Firmicutes and Proteobacteria-the two known dominant genus. Genus-level PCA (Principal component analysis) and OTU-level PCoA (Principal coordinates analysis) based on unweighted UniFrac distances showed that the bacterial community structure differed significantly between the samples from the 2 areas. 7 OTUs were detected in all samples, accounting for 4.4% of the total qualified assembly. Among the 7 OTUs, 3 OTUs were identified as Enterococcus, 2 OTUs were identified as Cronobacter, 1 OTU was identified as Bacillus and 1 OTU was identified as Alkaliphilus. Fifty-eight lactic acid bacteria (LAB) strains were isolated from the 20 koji samples with traditional microbial methods. Among them, Enterococcus faecium and Pediococcus pentosaceus were the dominant LAB isolates, with relative abundances of 51.72% and 31.03%. Despite the differences, a large number of shared bacteria were detected in samples from the two areas.

Duck tembusu virus (DTMUV) is newly emerged in poultry and causes great losses to the breeding industry in China and neighboring countries. Effective antiviral strategies are still being studied. Autophagy is a cellular degradative pathway, and our lab's previous data show that autophagy promotes DTMUV replication in vitro . To study the role of autophagy further in vivo , we utilized ducks as the animal model to investigate the autophagy responses in DTMUV-targeted tissues. And also, we utilized autophagy regulators, including Rapamycin (Rapa) as the autophagy enhancer, 3-Methyladenine (3-MA) and Chloroquine (CQ) as the autophagy inhibitors, to adjust the host autophagic levels and then study the effects of autophagy on tissue damages and virus replication. As a result, we first found DTMUV infection trigged autophagy and autophagy regulator treatments regulated autophagy levels successfully in duck spleens and brains. Next, we found that autophagy inhibitors inhibited DTMUV replication and alleviated DTMUV-induced pathological symptoms, whereas the autophagy inducer treatment led to the opposite effects. And we also found that autophagic regulation was correlated with the expression of innate immune genes, including pattern recognition receptors, type I interferons, and cytokines, and caused different effects in different tissues. In summary, we demonstrated that autophagy facilitated DTMUV replication, aggravated the developments of pathological symptoms and possibly counteracts the host's innate immunity response in vivo ., (Copyright © 2020 Hu, Pan, Cheng, Zhang, Wang, Chen, Zhu, Liu, Yang, Wu, Zhao, Huang, Zhang, Mao, Ou, Yu, Zhang, Liu, Tian, Pan, Rehman, Yin and Jia.)

Objectives: To date, very few hypervirulent and multiantibiotic-resistant bacterial strains have been reported. This study reports the first hypervirulent and multiantibiotic-resistant Pasteurella multocida sequence type 342 (ST342) strain (GH161213) isolated from a Pekin duck in China., Methods: Minimum inhibitory concentrations (MICs) were determined according to Clinical and Laboratory Standards Institute (CLSI) guidelines (VET01-A4, 2013). Determination of the P. multocida GH161213 median lethal dose (LD 50 ) was determined in a mouse model and in ducklings. Plasmid pRCAD0338PM-1 was transferred to Escherichia coli J53 Azr by conjugation. The whole genome sequence of P. multocida GH161213 was obtained using an Illumina HiSeq 2500 system. Antimicrobial resistance genes were analysed using the Comprehensive Antibiotic Resistance Database (CARD)., Results: Pasteurella multocida GH161213 is a hypervirulent strain with an LD 50 of <10 CFU in a mouse model and in ducklings. It also has a high level of multidrug resistance. Strain GH161213 contains a small conjugative plasmid harbouring the floR florfenicol resistance gene. It also contains multiple other antimicrobial resistance mechanisms., Conclusion: The genome sequence of P. multocida GH161213 reveals a multidrug-resistant genotype. This is the first reported hypervirulent and multiantibiotic-resistant P. multocida strain., (Copyright © 2019 International Society for Antimicrobial Chemotherapy. Published by Elsevier Ltd. All rights reserved.)

This study was conducted to determine the prevalence of integrons and associated gene cassettes in Escherichia coli (E. coli) isolates from waterfowls in Hainan, China. The antimicrobial resistance profile of the isolates was examined by using disc diffusion test. In addition, PCR, RFLP, plasmid replicon typing and DNA sequencing analyses were used for the characterization of integrase genes (class 1, 2 and 3) and associated gene cassettes. Approximatively, 90% of the isolates were positive for the integrase genes by PCR. Specifically, class 1 and class 2 integrons were found in 252 (81%) and 7 (2.3%) strains, respectively. While 21 (6.7%) isolates were positive for both class 1 and class 2 integrons. However, none of the isolate was positive for the class 3 integrons. In addition, 5 various cassette arrays, dfrA1-orfC, aadA2, aadA1, dfrA1-aadA1, and dfrA1-orfC- aadA1, were found within the variable regions (VRs) of class 1 integron isolates. While only single cassette array, dfrA1-sat2- aadA1, was identified within VRs of class 2 integron isolates. We identified incF plasmid as the most common plasmid type, which was detected in 81 of 243 VRs containing isolates. This study is the first report showing the baseline characteristics of integrons in E. coli isolates from waterfowls in Hainan, China. Our results provide evidence of the waterfowl birds as a reservoir of class 1 and class 2 integrons carrying antibiotic resistance gene cassettes. Therefore, strict preventive measures should be taken to avoid the spread of mobile genetic resistance elements in waterfowls in China., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Live attenuated vaccines are widely used to protect humans or animals from pathogen infections. We have previously developed a chicken embryo-attenuated Duck Hepatitis A Virus genotype 1 (DHAV-1) vaccine (CH60 strain). This study aims to understand the mechanisms that drive a virulent strain to an attenuated virus. Here, we systematically compared five DHAV-1 chicken embryo attenuated strains and 68 virulent strains. Phylogenetic analysis indicated that duck virulent strains isolated from different geographic regions of China undergo a convergent evolution in the chicken embryos. Comparative analysis indicated that the codon usage bias of the attenuated strains were shaped by chicken codons usage bias, which essentially contributed to viral adaption in the unsuitable host driven by incompatible translation. Of note, the missense mutations in coding region and mutations in untranslated regions may also contribute to viral attenuation of DHAV-1 to some extent. Importantly, we have experimentally confirmed that the expression levels of four viral proteins ( 2A 3 pro , 2A 3 pro , 3C pro , and 3D pro ) in the liver and kidney of ducks infected with an attenuated strain are significantly lower than that infected with a virulent strain, despite with similar virus load. Thus, the key mechanisms of viral attenuation revealed by this study may lead to innovative and easy approaches in designing live attenuated vaccines.

Background: Lethal Duck Enteritis Virus (DEV) infection can cause high morbidity and mortality of many species of waterfowl within the order Anseriformes. However, little is known about the function of viral genes including the conserved UL55 gene among alpha herpes virus due to the obstacles in maintenance and manipulation of DEV genome in host cells., Methods: In this paper, we constructed an infectious bacteria artificial chromosome (BAC) clone of the lethal clinical isolate duck enteritis virus Chinese virulent strain (DEV CHv) by inserting a transfer vector containing BAC mini-F sequence and selection marker EGFP into UL23 gene using homologous recombination. UL55 deletion and its revertant mutant were generated by two-step RED recombination in E. coli on basis of rescued recombinant virus. The function of UL55 gene in DEV replication and its effect on distribution of UL26.5 protein were carried out by growth characteristics and co-localization analysis., Results: The complete genome of DEV CHv can be stably maintained in E. coli as a BAC clone and reconstituted again in DEF cells. The generated UL55 deletion mutant based on DEV CHv-BAC-G displayed similar growth curves, plaque morphology and virus titer of its parental virus in infected Duck Embryo Fibroblast (DEF) cells. Immunofluorescence assay indicated that the loss of UL55 gene do not affect the distribution of UL26.5 protein in intracellular. These data also suggest infectious BAC clone of DEV CHv will facilitate the gene function studies of DEV genome., Conclusions: We have successfully developed an infectious BAC clone of lethal clinical isolate DEV CHv for the first time. The generated UL55 gene mutant based on that demonstrated this platform would be a very useful tool for functional study of DEV genes. We found the least known DEV UL55 is dispensable for virus replication and UL26.5 distribution, and it could be a very promise candidate locus for developing bivalent vaccine. Experiment are now in progress for testifying the possibility of UL55 gene locus as an exogenous gene insertion site for developing DEV vectored vaccine.