Some coronaviruses (CoVs) have an extra furin cleavage site (RRKR/S, furin-S2′ site) upstream of the fusion peptide in the spike protein, which plays roles in virion adsorption and fusion. Mutation ...of the S2′ site of QX genotype (QX-type) infectious bronchitis virus (IBV) spike protein (S) in a recombinant virus background results in higher pathogenicity, pronounced neural symptoms and neurotropism when compared with conditions in wild-type IBV (WT-IBV) infected chickens. In this study, we present evidence suggesting that recombinant IBV with a mutant S2′ site (furin-S2′ site) leads to higher mortality. Infection with mutant IBV induces severe encephalitis and breaks the blood–brain barrier. The results of a neutralization test and immunoprotection experiment show that an original serum and vaccine can still provide effective protection in vivo and in vitro. This is the first demonstration of IBV-induced neural symptoms in chickens with encephalitis and the furin-S2′ site as a determinant of neurotropism.
Swine enteric coronaviruses (SECoVs) including porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine deltacoronavirus (PDCoV), account for the majority of ...lethal watery diarrhea in neonatal pigs and pose significant economic and public health burdens in the world. While the three SECoVs primarily infect intestinal epithelia
and cause similar clinical signs, there are evident discrepancies in their cellular tropism and pathogenicity. However, the underlying mechanisms to cause the differences remain unclear. Herein, we employed porcine enteroids that are a physiologically relevant model of the intestine to assess the host epithelial responses following infection with the three SECoVs (PEDV, TGEV, and PDCoV). Although SECoVs replicated similarly in jejunal enteroids, a parallel comparison of transcriptomics datasets uncovered that PEDV and TGEV infection induced similar transcriptional profiles and exhibited a more pronounced response with more differentially expressed genes (DEGs) in jejunal enteroids compared with PDCoV infection. Notably, TGEV and PDCoV induced high levels of type I and III IFNs and IFN-stimulated gene (ISG) responses, while PEDV displayed a delayed peak and elicited a much lesser extent of IFN responses. Furthermore, TGEV and PDCoV instead of PEDV elicited a substantial upregulation of antigen-presentation genes and T cell-recruiting chemokines in enteroids. Mechanistically, we demonstrated that IFNs treatment markedly elevated the expression of NOD-like receptor (NLR) family NLRC5 and major histocompatibility complex class I (MHC-I) molecules. Together, our results indicate unique and common viral strategies for manipulating the global IFN responses and antigen presentation utilized by SECoVs, which help us a better understanding of host-SECoVs interactions.
Ornithobacterium rhinotracheale (ORT) has been associated with avian respiratory disease. On coinfection with other pathogens, ORT can cause serious health problems in avian species, leading to ...financial losses. To monitor the serologic prevalence of ORT in chicken flocks in China, 1,280 sera were collected to determine ORT antibodies among 64 flocks from 15 provinces of China using a commercial ELISA kit. The overall seroprevalence of ORT among the birds tested was 44.06%. In younger chickens, the serum positive rate was lower than that in older chickens, and with increased age, the serum positive rates increased. Older chickens had not only higher positive rates but also higher antibody levels. These data indicated that ORT infections were common in China. Because an ORT vaccine is currently not available, good disease management and biosecurity measures are required for effective disease control.
Opportunistic pathogens frequently cause volatile infections in hosts with compromised immune systems or a disrupted normal microbiota. The commensalism of diverse microorganisms contributes to ...colonization resistance, which prevents the expansion of opportunistic pathogens. Following microbiota disruption, pathogens promptly adapt to altered niches and obtain growth advantages. Nevertheless, whether and how resident bacteria modulate the growth dynamics of invasive pathogens and the eventual outcome of such infections are still unclear. Here, we utilized birds as a model animal and observed a resident bacterium exacerbating the invasion of Avibacterium paragallinarum (previously Haemophilus paragallinarum) in the respiratory tract. We first found that negligibly abundant Staphylococcus chromogenes, rather than Staphylococcus aureus, played a dominant role in Av. paragallinarum-associated infectious coryza in poultry based on epidemic investigations and in vitro analyses. Furthermore, we determined that S. chromogenes not only directly provides the necessary nutrition factor nicotinamide adenine dinucleotide (NAD+) but also accelerates its biosynthesis and release from host cells to promote the survival and growth of Av. paragallinarum. Last, we successfully intervened in Av. paragallinarum-associated infections in animal models using antibiotics that specifically target S. chromogenes. Our findings show that opportunistic pathogens can hijack commensal bacteria to initiate infection and expansion and suggest a new paradigm to ameliorate opportunistic infections by modulating the dynamics of resident bacteria.
•Full genomes of ten PPMV-1 isolates were determined in this study.•Ten PPMV-1 viruses belong to VI.1.2.2.2, VI.2.1.1.2.1, VI.2.1.1.2.2 or VII genotypes.•VI.1.2.2.2, VI.2.1.1.2.1 and VI.2.1.1.2.2 ...PPMV-1 viruses are circulating in pigeons in China.•VI.2.1.1.2.1 and VI.2.1.1.2.2 PPMV-1 viruses are pathogenic to pigeon.
Pigeon paramyxovirus type 1 (PPMV-1) is an antigenic variant of avian paramyxovirus type 1, which mainly infects pigeons. Here, we characterized ten PPMV-1 viruses isolated from pigeons in China during 1996–2019. Phylogenetic analysis of available complete genomes, F and HN genes of PPMV-1 from China showed that multiple PPMV-1 genotypes (I, II, VI, and VII) exist in pigeons in China. Ten PPMV-1 viruses isolated in this study belonged to genotypes VI.1.2.2.2, VI.2.1.1.2.1, VI.2.1.1.2.2 and VII respectively. Genotype VI is predominant in pigeons. VI.2.1.1.2.2 contains most recently isolated PPMV-1 viruses, suggesting that VI.2.1.1.2.2 is a prevalent genotype in pigeons in China. In vitro and in vivo studies showed that four representative viruses from genotypes VI.2.1.1.2.1 (TA14), VI.2.1.1.2.2 (SD19), VI.1.2.2.2 (SD16), and VII (JN08) could replicate efficiently in chicken embryo fibroblasts, while the replication titer of JN08 (VII) virus was significantly lower than that of VI gene viruses in pigeon embryo fibroblasts. The TA14 (VI.2.1.1.2.1) and SD19 (VI.2.1.1.2.2) viruses caused 20 % and 30 % mortality in pigeons, respectively. No birds infected with SD16 (VI.1.2.2.2) died during the study period. JN08 (VII) virus did not cause obvious clinical signs in infected pigeons. All data indicated that VI.2.1.1.2.2 is the prevalent genotype circulating in China and poses a major threat to pigeons, suggesting that a matched vaccine is necessary to control the disease.
•Pathogenicity of different passage of QX-like strain SZ were evaluated.•The complete genome mutations during continuous passage were analyzed.•This study is of great value to the control of IBV in ...poultry industry.
Infectious bronchitis (IB) remains a major problem in the global poultry industry despite the many available vaccines. Live attenuated vaccines are the most effective means of preventing IB and are traditionally generated by serial passaging of a wild strain in embryonated chicken eggs. In this study, the SZ isolate of the QX-like infectious bronchitis virus (IBV) was continuously passaged in chicken embryos for 250 passages. We compared the pathogenicity of different passages (SZ50, SZ100, SZ150, SZ200 and SZ250) of strain SZ by clinical signs, gross lesions, viral load, tissue tropism, weight gain and tracheal ciliary activity. As the passaging increased in the chicken embryos, the strain lost its ability to infect many organs, and the viral pathogenicity gradually decreased. We also found 23 genomic variations of the QX-like strain SZ throughout the passaging process by further analyzing its complete genome sequence. This work offers valuable insight for IBV vaccine development and further research on the IBV attenuation mechanisms.
Marek’s disease virus (MDV), reticuloendotheliosis virus (REV), avian reovirus (ARV), chicken infectious anemia virus (CIAV), infectious bursal disease virus (IBDV), and fowl adenovirus (FAdV) are ...important causes of disease in poultry. To investigate the infection status of the above six viruses in chickens in China, 1,187 samples from chicken flocks were collected and tested using a newly developed multiplex reverse-transcription quantitative real-time PCR (MRT-qPCR) assay in the study. A series of validation tests confirmed that the MRT-qPCR assay has high specificity, sensitivity, and repeatability. As for six detected pathogens, CIAV had the highest detection ratio, while ARV was not detected in any samples. In the spleen samples, the coinfection rate for MDV and CIAV was 1.6%, and that for REV and CIAV was 0.4%. In the bursa samples, the coinfection rate for FAdV and CIAV was 0.3%, and that for IBDV and CIAV was 1%. In the thymus samples, the coinfection rates for MDV and CIAV and for REV and CIAV were both 0.8%. Our study indicates that the coinfection of these viruses was existing in chickens in China. Through the detection of clinical samples, this study provides data on the coinfections of the above six pathogens and provides a basis for the further study of viral coinfection in chickens.
The taro harvesting process is affected by a complex system composed of particle mechanics system and multi-body dynamics system. The discrete element method(DEM) can effectively solve the nonlinear ...problem of the interaction between harvesting components and working materials. Therefore, the discrete element model of taro tiller plants is of great importance for taro harvesting. This paper proposes a simulation method to establish a discrete element flexible plant model and dynamic clamping and pulling process of taro tiller plant. Discrete Element models of taro corm and flexible tiller petiole and leaf were established using DEM method, and the discrete element flexible model of the taro plant was established. Taro clamping and pulling force testing platform was designed and built. The single factor and Plackett-Burman experiments were used to determine the simulation parameters and optimize the taro plant model by taking the correlation coefficient of clamping force and correlation coefficient of pulling force collected from the simulation and the bench experiment as the experiment index. The parameter calibration results of discrete element model of taro plant are as follows: petiole-petiole method/tangential contact stiffness was 8.15×10
9
N·m
-3
, and normal/tangential critical stress was 6.65×10
6
Pa. The contact stiffness of pseudostem- corm method was 1.22×10
9
N·m
-3
, the critical stress of normal/tangential was 1.18×10
5
Pa, and the energy of soil surface was 4.15×10
6
J·m
-3
. When the pulling speed is 0.1, 0.2, 0.3, 0.4 and 0.5 m·s
-1
, the correlation coefficients between the simulation experiment and the bench experiment are 0.812, 0.850, 0.770, 0.697 and 0.652, respectively. The average value of correlation coefficient is 0.756, indicating that the simulated discrete element plant model is close to the real plant model. The discrete element model of taro plant established in this paper has high reliability. The final purpose of this paper is to provide a model reference for the design and optimization of taro harvester by discrete element method.
Newcastle disease virus (NDV) is a highly infectious viral disease, which can affect chickens and many other kinds of birds. The main virulence factor of NDV, the fusion (F) protein, is located on ...the viral envelope and plays a major role in the virus' ability to penetrate cells and cause host cell fusion during infection. Multiple highly conserved tyrosine and di-leucine (LL) motifs in the cytoplasmic tail (CT) of the virus may contribute to F protein functionality in the viral life cycle.
To examine the contribution of the LL motif in the biosynthesis, transport, and function of the F protein, we constructed and rescued a NDV mutant strain, rSG10*-F/L537A, with an L537A mutation using a reverse genetic system. Subsequently, we compared the differences in the syncytium formation ability, pathogenicity, and replication levels of wild-type rSG10* and the mutated strain.
Compared with rSG10*, rSG10*-F/L537A had attenuated syncytial formation and pathogenicity, caused by a viral budding defect. Further studies showed that the LL-motif mutation did not affect the replication, transcription, or translation of the virus genome but affected the expression of the F protein at the cell surface.
We concluded that the LL motif in the NDV F CT affected the regulation of F protein expression at the cell surface, thus modulating the viral fusion ability and pathogenic phenotype.
Coronary artery spasm (CAS) plays an important role in the pathogeneses of various ischemic heart diseases and has gradually become a common cause of life-threatening arrhythmia. The specific ...molecular mechanism of CAS has not been fully elucidated, nor are there any specific diagnostic markers for the condition. Therefore, this study aimed to examine the specific molecular mechanism underlying CAS, and screen for potential diagnostic markers. To this end, we successfully constructed a rat CAS model and achieved in vitro culture of a human coronary-artery smooth-muscle cell (hCASMC) contraction model. Possible molecular mechanisms by which protein kinase C (PKC) regulated CAS through the C kinase-potentiated protein phosphatase 1 inhibitor of 17 kDa (CPI-17)/myosin II regulatory light chain (MLC2) pathway were studied in vivo and in vitro to screen for potential molecular markers of CAS. We performed hematoxylin and eosin staining, myocardial zymogram, and transmission electron microscopy to determine myocardial and coronary artery injury in CAS rats. Then, using immunohistochemical staining, immunofluorescence staining, and Western blotting, we further demonstrated a potential molecular mechanism by which PKC regulated CAS via the CPI-17/MLC2 pathway. The results showed that membrane translocation of PKCα occurred in the coronary arteries of CAS rats. CPI-17/MLC2 signaling was observably activated in coronary arteries undergoing CAS. In addition, in vitro treatment of hCASMCs with angiotensin II (Ang II) increased PKCα membrane translocation while consistently activating CPI-17/MLC2 signaling. Conversely, GF-109203X and calphostin C, specific inhibitors of PKC, inactivated CPI-17/MLC2 signaling. We also collected the coronary artery tissues from deceased subjects suspected to have died of CAS and measured their levels of phosphorylated CPI-17 (p-CPI-17) and MLC2 (p-MLC2). Immunohistochemical staining was positive for p-CPI-17 and p-MLC2 in the tissues of these subjects. These findings suggest that PKCα induced CAS through the CPI-17/MLC2 pathway; therefore, p-CPI-17 and p-MLC2 could be used as potential markers for CAS. Our data provide novel evidence that therapeutic strategies against PKC or CPI-17/MLC2 signaling might be promising in the treatment of CAS.
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