Entire genome sequence analysis of genotype IX Newcastle disease viruses reveals their early-genotype phylogenetic position and recent-genotype genome size

<p>Abstract</p> <p>Background</p> <p>Six nucleotide (nt) insertion in the 5'-noncoding region (NCR) of the nucleoprotein (NP) gene of Newcaslte disease virus (NDV) is considered to be a genetic marker for recent genotypes of NDV, which emerged after 1960. However, F48-like NDVs from China, identified a 6-nt insert in the NP gene, have been previously classified into genotype III or genotype IX.</p> <p>Results</p> <p>In order to clarify their phylogenetic position and explore the origin of NDVs with the 6-nt insert and its significance in NDV evolution, we determined the entire genome sequences of five F48-like viruses isolated in China between 1946 and 2002 by RT-PCR amplification of overlapping fragments of full-length genome and rapid amplification of cDNA ends. All the five NDV isolates shared the same genome size of 15,192-nt with the recent genotype V-VIII viruses whereas they had the highest homology with early genotype III and IV isolates.</p> <p>Conclusions</p> <p>The unique characteristic of the genome size and phylogenetic position of F48-like viruses warrants placing them in a separate geno-group, genotype IX. Results in this study also suggest that genotype IX viruses most likely originate from a genotype III virus by insertion of a 6-nt motif in the 5'-NCR of the NP gene which had occurred as early as in 1940 s, and might be the common origin of genotype V-VIII viruses.</p

Newcastle disease virus induces testicular damage and disrupts steroidogenesis in specific pathogen free roosters

Newcastle disease (ND), which is caused by Newcastle disease virus (NDV), can cause heavy economic losses to the poultry industry worldwide. It is characterised by extensive pathologies of the digestive, respiratory, and nervous systems and can cause severe damage to the reproductive system of egg-laying hens. However, it is unknown whether NDV replicates in the male reproductive system of chickens and induces any pathologies. In this study, we selected a representative strain (i.e. ZJ1) of the most common genotype (i.e. VII) of NDV to investigate whether NDV can induce histological, hormonal, and inflammatory responses in the testes of specific pathogen free (SPF) roosters. NDV infection increased the expression of toll like receptor TLR3, TLR7, MDA5, IFN-α, IFN-β, IFN-γ, IL-8, and CXCLi1 in the testes of NDV-infected roosters at 5 days post-infection (dpi). Severe histological changes, including decrease in the number of Sertoli cells and individualized, shrunken spermatogonia with pyknotic nuclei, were observed at 3 dpi. At 5 dpi, the spermatogenic columns were disorganized, and there were fewer cells, which were replaced by necrotic cells, lipid vacuoles, and proteinaceous homogenous material. A significant decrease in the plasma concentrations of testosterone and luteinizing hormone (LH) and the mRNA expression of their receptors in the testes, steroidogenic acute regulatory protein, cytochrome P450 side-chain cleavage enzyme, and 3β-hydroxysteroid dehydrogenase in the NDV-infected group was observed relative to those in the control group (P < 0.05). Collectively, these results indicate that NDV infection induces a severe inflammatory response and histological changes, which decrease the steroidogenesis. © 2020 The Author(s)

Pathobiology of Avian avulavirus 1:special focus on waterfowl

Avian avulaviruses serotype 1 (abbreviated as APMV-1 for the historical name avian paramyxovirus 1) are capable of infecting a wide spectrum of avian species with variable clinical symptoms and outcomes. Ease of transmission has allowed the virus to spread worldwide with varying degrees of virulence depending upon the virus strain and host species. The emergence of new virulent genotypes from global epizootics, and the year-to-year genomic changes in low and high virulence APMV-1 imply that distinct genotypes of APMV-1 are simultaneously evolving at different geographic locations across the globe. This vast genomic diversity may be favoured by large variety of avian species susceptibility to APMV-1 infection, and by the availability of highly mobile wild birds. It has long been considered that waterfowls are not sensitive to APMV-1 and are unable to show any clinical signs, however, outbreaks from the 90's contradict these concepts. The APMV-1 isolates are increasingly reported from the waterfowl. Waterfowl have strong innate immune responses, which minimize the impact of virus infection, however, are unable to prevent the viral shedding. Numerous APMV-1 are carried by domestic waterfowl intermingling with terrestrial poultry. Therefore, commercial ducks and geese should be vaccinated against APMV-1 to minimize the virus shedding and for the prevention the transmission. Genetic diversity within APMV-1 demonstrates the need for continual monitoring of viral evolution and periodic updates of vaccine seed-strains to achieve efficient control and eradication of APMV-1 in waterfowls

Potential of genotype VII Newcastle disease viruses to cause differential infections in chickens and ducks

Newcastle disease (ND), caused by ND virus (NDV), is one of the most infectious and economically important diseases of the poultry industry worldwide. While infections are reported in a wide range of avian species, the pathogenicity of chicken-origin virulent NDV isolates in ducks remains elusive. In this study, two NDV strains were isolated and biologically and genetically characterized from an outbreak in chickens and apparently healthy ducks. Pathogenicity assessment indices, including the mean death time (MDT), intracerebral pathogenicity index (ICPI) and cleavage motifs in the fusion (F) protein, indicated that both isolates were velogenic in nature. While these isolates carried pathogenic characteristics, interestingly they showed differential pathogenicity in ducks. The chicken-origin isolate caused high (70%) mortality, whereas the duck-origin virus resulted in low (20%) mortality in 4-week-old ducks. Intriguingly, both isolates showed comparable disease pathologies in chickens. Full-genome sequence analysis showed that the virus genome contains 15 192 nucleotides and carried features that are characteristic of velogenic strains of NDV. A phylogenetic analysis revealed that both isolates clustered in class II and genotype VII. However, there were several mutations in the functionally important regions of the fusion (F) and haemagglutinin-neuraminidase (HN) proteins, which may be responsible for the differential pathogenicity of these viruses in ducks. In summary, these results suggest that NDV strains with the same genotype show differential pathogenicity in chickens and ducks. Furthermore, chicken-origin virulent NDVs are more pathogenic for ducks than duck-origin viruses. These findings propose a role for chickens in the evolution of viral pathogenicity and the potential genetic resistance of ducks to poultry viruses

Supplementation of Vitamin E Protects Chickens from Newcastle Disease Virus-Mediated Exacerbation of Intestinal Oxidative Stress and Tissue Damage

BACKGROUND/AIMS: Newcastle disease virus (NDV) causes a highly devastating and contagious disease in poultry, which is mainly attributed to extensive tissue damages in the digestive, respiratory and nervous systems. However, nature and dynamics of NDV-induced oxidative stresses in the intestine of chickens remain elusive. METHODS: In this study, we examined the magnitude of intestinal oxidative stress and histopathological changes caused by the virulent NDV infection, and explored the protective roles of vitamin E (vit. E) in ameliorating these pathological changes. For these purposes, chickens were divided into four groups namely i) non supplemented and non-challenged (negative control, CON); ii) no supplementation of vit. E but challenged with ZJ1 (positive control, NS+CHA); iii) vit. E supplementation at the dose of 50 IU/day/Kg body weight and ZJ1 challenge (VE50+CHA); and 4) vit. E supplementation at the dose of 100 IU/day/Kg body weight and ZJ1 challenge (VE100+CHA). In all groups, we analyzed concentrations of glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (T-AOC), and activity of glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) using biochemical methods. The virus loads were determined by quantitative RT-PCR and antibody titers by hemagglutination inhibition assays. We also examined the histopathological changes in the duodenal and jejunal mucosa at 3 and 5-day post infection (dpi) with NDV. RESULTS: A significant elevation in the NO level was observed in NDV challenged chickens compared to the CON chickens at 2 dpi. The MDA contents were significantly increased whereas GSH was significantly decreased in NDV-challenged chickens compared to control. Furthermore, activities of GST, CAT, SOD, as well as the TOAC were markedly decreased in challenged chickens in comparison with control. Virus copy numbers were higher in NDV infected NS+CHA group compared to other groups. Severe histopathological changes including inflammation, degeneration and broken villi were observed in the intestine of NDV challenged chickens. However, all these malfunctions of antioxidant system and pathological changes in the intestine were partially or completely reversed by the vit. E supplementation. CONCLUSIONS: Our results suggest that NDV infection causes oxidative stress and histopathological changes in the duodenum and jejunum of chickens, which can be partially or fully ameliorated by supplementation of vit. E. Additionally, these findings suggest that oxidative stress contributes to the intestinal damages in NDV infected chickens. These findings will help to understand the pathogenesis of NDV and further investigation of therapeutic agents for control of Newcastle disease

Production, characterization, and epitope mapping of a monoclonal antibody against genotype VII Newcastle disease virus V protein

Newcastle disease virus (NDV) V protein is crucial for viral interferon (IFN) antagonism and virulence, determining its host range restriction. However, little information is available on the B cell epitopes of V protein and the subcellular movement of V protein in the process of NDV infection. In this study, the monoclonal antibody (mAb) clone 3D7 against genotype VII NDV V protein was generated by immunizing mice with a purified recombinant His-tagged carboxyl-terminal domain (CTD) region of V protein. Fine epitope mapping analysis and B-cell epitope prediction indicated that mAb 3D7 recognized a linear epitope 152RGPAELWK159, which is located in the V protein CTD region. Sequence alignment showed that the mAb clone 3D7-recognized epitope is highly conserved among Class II genotype VII NDV strains, but not among other genotypes, suggesting it could serve as a genetic marker to differentiate NDV genotypes. Furthermore, the movement of V protein during NDV replication in infected cells were determined by using this mAb. It was found that V protein localized around the nucleus during virus replication. The establishment of V protein-specific mAb and identification of its epitope extend our understanding of the antigenic characteristics of V protein and provide a basis for the development of epitope-based diagnostic assays

Prosedur penyelesaian pembiayaan bermasalah pada akad mudharabah dalam rangka meminimalisir resiko di BMT Amanah Usaha Mulia Magelang

Permasalah kehidupan perekonomian yang sulit, membuat masyarakat berinisiatif untuk membuka usaha sendiri. Mereka membutuhkan suatu bantuan berupa dana untuk memperlancar usahanya, maka BMT Amanah Usaha Mulia Magelang ikut untuk mengembangkan produknya yaitu pembiayaan mudharabah sesuai perkembangan dunia perbankan dalam target peningkatan keuntungan dan menyejahterakan masyarakat. Dengan diberikanya pembiayaan tersebut, terkadang muncul adanya pembiayaan bermasalah dikarenakan ada beberapa faktor diantaranya ketidakmampuan anggota untuk membayar tepat waktu atau jatuh tempo pembayaran diakibatkan karena usaha anggota yang kurang lancar dan lain sebagaianya. Tugas Akhir ini berjudul “ Prosedur Penyelesaian Pembiayaan Bermasalah pada Akad Mudharabah Dalam Rangka Meminimalisir Risiko” Berdasarkan judul tersebut dapat diambil rumusan masalah yaitu apa penyebab terjadinya pembiayaan bermasalah pada BMT Amanah Usaha Mulia Magelang dan bagaimana prosedur penyelesaian pembiayaaan bermasalah pada akad mudharabah di BMT Amanah Usaha Mulia Magelang. Penelitian ini merupakan penelitian lapangan dimana sumber data yang digunakan berasal dari data primer dan sekunder yang diperoleh melalui metode wawancara dengan manajer, bagian pembiayaan dan dokumentasi. Metode yang digunakan dalam penelitian ini adalah deskriptif kualitatif yang bertujuan untuk menggambarkan secara sistematis dan akurat mengenai objek penelitian. Berdasarkan hasil penelitian dapat disimpulkan bahwa penyebab terjadinya pembiayaan bermasalah yaitu faktor internal meliputi kurang telitinya petugas BMT dalam menganalisi data calon anggota, kurang disiplinya dalam penagihan dan eksternal meliputi karakter anggota yang kurang baik, usahanya bangkrut dan terjadinya bencana alam yang tidak terduga. Adapun prosesdur yang digunakan BMT Amanah Usaha Mulia dalam menyelesaian pembiayaan bermasalah pada akad mudharabah dengan cara kekeluargaan atau musyawarah dengan anggota, penjadwalan kembali (rescheduling), persyaratan kembali (reconditioning), pengambilan jaminan (eksekusi), dan write off final. Di BMT Amanah Usaha Mulia dalam penyelesaian pembiayaan bermasalah jarang menngunakan jalur hukum, tetapi sering menggunakan cara kekeluargaan yang dianggap lebih efektif dan eksekusi jaminan apabila anggota tersebut sudah mengalami macet atau bermasalah

Analysis on the Changes of Agro-Meteorological Thermal Indices in Northeast China under RCP4.5 Scenario Using the PRECIS2.1

As a main grain production area in China, Northeast China (NEC) is highly influenced by the higher warming trend than elsewhere in China or even the globe. As the warming trend goes on, scientific understanding of the changes of thermal conditions in NEC will be essential for taking agricultural adaptation measures. In this paper, the high-resolution (25 km) corrected outputs of PRECIS (Providing Regional Climates for Impacts Studies) under the Representative Concentration Pathway 4.5 (RCP4.5) scenario were used to analyze the changes of the agro-climatic thermal resources in NEC. Results showed that accumulated temperature ≥10 °C (AT10) could increase by 300–500 °C·day, 500–900 °C·day, and 750–1000 °C·day in 2011–2040 (2020s), 2041–2070 (2050s), and 2071–2100 (2080s), respectively, relative to the baseline period of 1961–1990. The potential growing season (PGS) would then be prolonged by 11–19 days, 23–35 days, and 25–37 days in 2020s, 2050s and 2080s, respectively, compared to baseline. The spatial features for the changes of thermal indices would show a large increase of AT10 and an extension of PGS and Frost-free period (FFP) in mountainous areas compared to plains; the increment of AT10 would be greater in southern NEC than that in the north, with over 1000 °C·day in the southern parts and 700–800 °C·day in the northern parts in the 2080s. Additionally, the three thermal indices would increase rapidly from 2020s to 2050s compared to the period from 2050s to the 2080s, coincides with the greenhouse gas concentrations peak around the mid-period of the 21st century, and the decline towards the end of the 21st century under RCP4.5 forcing. The results of this paper could act as a guide to taking advantage of increasing thermal resources in NEC, and would be helpful for local practitioners in decision-making

Pathogenicity and immunogenicity of gI/gE/TK-gene-deleted Felid herpesvirus 1 variants in cats

Abstract Background Felid herpesvirus 1 (FHV-1) is a major pathogenic agent of upper respiratory tract infections and eye damage in felines worldwide. Current FHV-1 vaccines offer limited protection of short duration, and therefore, do not reduce the development of clinical signs or the latency of FHV-1. Methods To address these shortcomings, we constructed FHV ∆gIgE-eGFP, FHV ∆TK mCherry, and FHV ∆gIgE/TK eGFP-mCherry deletion mutants (ΔgI/gE, ΔTK, and ΔgIgE/TK, respectively) using the clustered regularly interspaced palindromic repeats (CRISPR)/CRISP-associated protein 9 (Cas9) system (CRISPR/Cas9), which showed safety and immunogenicity in vitro. We evaluated the safety and efficacy of the deletion mutants administered with intranasal (IN) and IN + subcutaneous (SC) vaccination protocols. Cats in the vaccination group were vaccinated twice at a 4-week interval, and all cats were challenged with infection 3 weeks after the last vaccination. The cats were assessed for clinical signs, nasal shedding, and virus-neutralizing antibodies (VN), and with postmortem histological testing. Results Vaccination with the gI/gE-deleted and gI/gE/TK-deleted mutants was safe and resulted in significantly lower clinical disease scores, fewer pathological changes, and less nasal virus shedding after infection. All three mutants induced virus-neutralizing antibodies after immunization. Conclusions In conclusion, this study demonstrates the advantages of FHV-1 deletion mutants in preventing FHV-1 infection in cats