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Significant mortalities of racing pigeons occurred in Australia in late 2011 associated with a pigeon paramyxovirus serotype 1 (PPMV-1) infection. The causative agent, designated APMV-1/pigeon/Australia/3/2011 (P/Aus/3/11), was isolated from diagnostic specimens in specific pathogen free (SPF) embryonated eggs and was identified by a Newcastle Disease virus (NDV)-specific RT-PCR and haemagglutination inhibition (HI) test using reference polyclonal antiserum specific for NDV. The P/Aus/3/11 strain was further classified as PPMV-1 using the HI test and monoclonal antibody 617/161 by HI and phylogenetic analysis of the fusion gene sequence. The isolate P/Aus/3/11 had a slow haemagglutin-elution rate and was inactivated within 45 min at 56 °C. Cross HI tests generated an R value of 0.25, indicating a significant antigenic difference between P/Aus/3/11 and NDV V4 isolates. The mean death time (MDT) of SPF eggs infected with the P/Aus/3/11 isolate was 89.2 hr, characteristic of a mesogenic pathotype, consistent with other PPMV-1 strains. The plaque size of the P/Aus/3/11 isolate on chicken embryo fibroblast (CEF) cells was smaller than those of mesogenic and velogenic NDV reference strains, indicating a lower virulence phenotype in vitro and challenge of six-week-old SPF chickens did not induce clinical signs. However, sequence analysis of the fusion protein cleavage site demonstrated an 112RRQKRF117 motif, which is typical of a velogenic NDV pathotype. Phylogenetic analysis indicated that the P/Aus/3/11 isolate belongs to a distinct subgenotype within class II genotype VI of avian paramyxovirus type 1. This is the first time this genotype has been detected in Australia causing disease in domestic pigeons and is the first time since 2002 that an NDV with potential for virulence has been detected in Australia.

Newcastle disease (ND) is one of the most economically devastating infectious diseases affecting the poultry industry. Virulent Newcastle disease virus (NDV) can cause high mortality and severe tissue lesions in the respiratory, gastrointestinal, neurological, reproductive and immune systems of poultry. Tremendous progress has been made in preventing morbidity and mortality caused by ND based on strict biosecurity and wide vaccine application. In recent decades, the continual evolution of NDV has resulted in a total of twenty genotypes, and genetic variation may be associated with disease outbreaks in vaccinated chickens. In some countries, the administration of genotype-matched novel vaccines in poultry successfully suppresses the circulation of virulent NDV strains in the field. However, virulent NDV is still endemic in many regions of the world, especially in low- and middle-income countries, impacting the livelihood of millions of people dependent on poultry for food. In ND-endemic countries, although vaccination is implemented for disease control, the lack of genotype-matched vaccines that can reduce virus infection and transmission as well as the inadequate administration of vaccines in the field undermines the effectiveness of vaccination. Dissection of the profiles of existing ND vaccines is fundamental for establishing proper vaccination regimes and developing next-generation vaccines. Therefore, in this article, we provide a broad review of commercial and experimental ND vaccines and promising new platforms for the development of next-generation vaccines.

The control of poultry diseases has relied heavily on the use of many live and inactivated vaccines. However, over the last 30 yr, recombinant DNA technology has been used to generate many novel poultry vaccines. Fowlpox virus and turkey herpesvirus are the two main vectors currently used to construct recombinant vaccines for poultry. With the use of these two vectors, more than 15 recombinant viral vector vaccines against Newcastle disease, infectious laryngotracheitis, infectious bursal disease, avian influenza, and Mycoplasma gallisepticum have been developed and are commercially available. This review focuses on current knowledge about the safety and efficacy of recombinant viral vectored vaccines and the mechanisms by which they facilitate the control of multiple diseases. Additionally, the development of new recombinant vaccines with novel vectors will be briefly discussed.

Newcastle disease is caused by virulent strains of Newcastle disease virus (NDV), which causes substantial morbidity and mortality events worldwide in poultry. The virus strains can be differentiated as lentogenic, mesogenic, or velogenic based on a mean death time in chicken embryos. Currently, velogenic strains of NDV are not endemic in United States domestic poultry; however, these strains are present in other countries and are occasionally detected in wild birds in the U.S. A viral introduction into domestic poultry could have severe economic consequences due to the loss of production from sick and dying birds, the cost of control measures such as depopulation and disinfection measures, and the trade restrictions that would likely be imposed as a result of an outbreak. Due to the disease-free status of the U.S. and the high cost of a potential viral incursion to the poultry industry, a qualitative risk analysis was performed to evaluate the vulnerabilities of the U.S. against the introduction of virulent strains of NDV. The most likely routes of virus introduction are explored and data gathered by several federal agencies is provided. Recommendations are ultimately provided for data that would be useful to further understand NDV on the landscape and to utilize all existing sampling opportunities to begin to comprehend viral movement and further characterize the risk of NDV introduction into the U.S.

Newcastle disease (ND) has been a consistent risk factor to the poultry industry worldwide. Its pathogen, Newcastle disease virus (NDV), is also a promising antitumor treatment candidate. The pathogenic mechanism has intrigued the great curiosity of researchers, and advances in the last two decades have been summarized in this paper. The NDV’s pathogenic ability is highly related to the basic protein structure of the virus, which is described in the Introduction of this review. The overall clinical signs and recent findings pertaining to NDV-related lymph tissue damage are then described. Given the involvement of cytokines in the overall virulence of NDV, cytokines, particularly IL6 and IFN expressed during infection, are reviewed. On the other hand, the host also has its way of antagonizing the virus, which starts with the detection of the pathogen. Thus, advances in NDV’s physiological cell mechanism and the subsequent IFN response, autophagy, and apoptosis are summarized to provide a whole picture of the NDV infection process.

Due to their large population sizes, synanthropic birds, including native and invasive species, can transmit pathogens to other vertebrates, and even humans. In particular, Newcastle disease virus (NDV) can cause lethal infections in a wide range of avian species as well as mild infections in humans and other non-avian hosts. In this study conducted in Barcelona, Spain, we assessed the seroprevalence of NDV in three synanthropic species comprising one native species (feral pigeon, Columba livia var. domestica ; N = 16) and two invasive parrots (monk parakeet, Myiopsitta monachus , N = 50; and rose-ringed parakeet, Psittacula krameri , N = 23). These species are considered to have played important roles in the introduction and spread of NDV due to their high population densities, gregariousness, and prolonged viral shedding through faeces. We found a high seroprevalence of NDV in the feral pigeon (56%) and monk parakeet (38%) but a low seroprevalence in the rose-ringed parakeet (4%). These differences may be explained by the frequent interactions among feral pigeons and monk parakeets in their daily lives facilitating viral transmission from the former to the latter, because the feral pigeon is a well-known NDV reservoir. By contrast, the low seroprevalence of NDV in rose-ringed parakeets could be explained by its less frequent contacts with other urban bird species. The high seroprevalence of NDV in feral pigeons and monk parakeets provides new insights into the roles of novel interspecific relationships in the transmission and spread of NDV, and the risk of these synanthropic species as a source of NDV in urban environments. Finally, our findings highlight the need to monitor NDV in both native and non-native birds to prevent its spread to poultry, wildlife, and humans.

Cell culture-based production of vector-based vaccines and virotherapeutics is of increasing interest. The vectors used not only retain their ability to infect cells but also induce robust immune responses. Using two recombinant vesicular stomatitis virus (rVSV)-based constructs, we performed a proof-of-concept study regarding an integrated closed single-use perfusion system that allows continuous virus harvesting and clarification. Using suspension BHK-21 cells and a fusogenic oncolytic hybrid of vesicular stomatitis virus and Newcastle disease virus (rVSV-NDV), a modified alternating tangential flow device (mATF) or tangential flow depth filtration (TFDF) systems were used for cell retention. As the hollow fibers of the former are characterized by a large internal lumen (0.75 mm; pore size 0.65 μm), membrane blocking by the multi-nucleated syncytia formed during infection could be prevented. However, virus particles were completely retained. In contrast, the TFDF filter unit (lumen 3.15 mm, pore size 2–5 μm) allowed not only to achieve high viable cell concentrations (VCC, 16.4–20.6×10 6 cells/mL) but also continuous vector harvesting and clarification. Compared to an optimized batch process, 11-fold higher infectious virus titers were obtained in the clarified permeate (maximum 7.5×10 9 TCID 50 /mL). Using HEK293-SF cells and a rVSV vector expressing a green fluorescent protein, perfusion cultivations resulted in a maximum VCC of 11.3×10 6 cells/mL and infectious virus titers up to 7.1×10 10 TCID 50 /mL in the permeate. Not only continuous harvesting but also clarification was possible. Although the cell-specific virus yield decreased relative to a batch process established as a control, an increased space-time yield was obtained. Key points • Viral vector production using a TFDF perfusion system resulted in a 460% increase in space-time yield • Use of a TFDF system allowed continuous virus harvesting and clarification • TFDF perfusion system has great potential towards the establishment of an intensified vector production

Heat stress, one of the critical obstacles to poultry sector in subtropical and tropical countries, reduces performance, immune response, and animal welfare. This study examined the effect of dietary inclusion of probiotic (PRO), citric acid (CIT), garlic powder (GAR) or their combinations on growth, blood constituents, ileal microflora and morphology and humoral immunity of broiler chickens subjected to cyclic heat stress. Four hundred ninety one-day-old Ross-308 broiler chicks were randomly allocated to 7 groups with 7 replicates of 10 birds each as follows: control (C) group received the basal diet without supplements, PRO, CIT and GAR groups supplemented with 0.5 g kg−1 multi-strain probiotic mixture (MPM), citric acid and garlic powder, respectively. PRO-CIT and PRO-GAR groups treated with 0.5 g kg−1 MPM, and 0.5 g kg−1 citric acid and garlic powder, while CIT-GAR group fed diet with 0.5 g kg−1 of citric acid and garlic powder. Results revealed that dietary supplements and their combinations improved (P < 0.001) growth performance and decreased abdominal fat of heat-stressed birds. Dietary supplements decreased (P < 0.01) serum concentrations of cholesterol, triglycerides and LDL, while HDL was elevated (P < 0.05). Feed additives reduced (P < 0.01) ileal enumeration of Escherichia coli and total coliform while Lactobacillus count was increased (P < 0.05) only in MPM-enriched groups. Supplementation of these natural products improved (P < 0.01) ileal architecture while humoral immune response was not significantly influenced except antibody titre against Newcastle disease virus which was increased (P < 0.05) in MPM-supplemented groups. Conclusively, addition of the dietary supplements and their combinations, particularly, probiotic and citric acid combination can improve productive performance, and intestinal flora and histomorphometry of broilers exposed to cyclic heat stress.

•Multivalent vHVT vaccines protect against HPAIVs, IBDV, and NDV challenges.•HVT COBRA H5 insert alone or in combination protects against diverse Gs/GD H5 HPAIV.•Vaccines against HPAIV provided clinical protection and reduced viral shedding. Vaccines are an essential tool for the control of viral infections in domestic animals. We generated recombinant vector herpesvirus of turkeys (vHVT) vaccines expressing computationally optimized broadly reactive antigen (COBRA) H5 of avian influenza virus (AIV) alone (vHVT-AI) or in combination with virus protein 2 (VP2) of infectious bursal disease virus (IBDV) (vHVT-IBD-AI) or fusion (F) protein of Newcastle disease virus (NDV) (vHVT-ND-AI). In vaccinated chickens, all three vHVT vaccines provided 90–100% clinical protection against three divergent clades of high pathogenicity avian influenza viruses (HPAIVs), and significantly decreased number of birds and oral viral shedding titers at 2 days post-challenge compared to shams. Four weeks after vaccination, most vaccinated birds had H5 hemagglutination inhibition antibody titers, which significantly increased post-challenge. The vHVT-IBD-AI and vHVT-ND-AI vaccines provided 100% clinical protection against IBDVs and NDV, respectively. Our findings demonstrate that multivalent HVT vector vaccines were efficacious for simultaneous control of HPAIV and other viral infections.

Background Avian influenza (AI) is a disease caused by the avian influenza virus (AIV). These viruses spread naturally among wild aquatic birds worldwide and infect domestic poultry, other birds, and other animal species. Currently, real-time reverse transcription polymerase chain reaction (rRT-PCR) is mainly used to detect the presence of pathogens and has good sensitivity and specificity. However, the diagnosis requires sophisticated instruments under laboratory conditions, which significantly limits point-of-care testing (POCT). Rapid, reliable, non-lab-equipment-reliant, sensitive, and specific diagnostic tests are urgently needed for rapid clinical detection and diagnosis. Our study aimed to develop a reverse transcription recombinase polymerase amplification (RT-RPA)/CRISPR method which improves on these limitations. Methods The Cas12a protein was purified by affinity chromatography with Ni-agarose resin and observed using sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). Specific CRISPR RNA (crRNA) and primers targeting the M and NP genes of the AIV were designed and screened. By combining RT-RPA with the Cas12a/crRNA trans-cleavage system, a detection system that uses fluorescence readouts under blue light or lateral flow strips was established. Sensitivity assays were performed using a tenfold dilution series of plasmids and RNA of the M and NP genes as templates. The specificity of this method was determined using H1–H16 subtype AIVs and other avian pathogens, such as newcastle disease virus (NDV), infectious bursal disease virus (IBDV), and infectious bronchitis virus (IBV). Results The results showed that the method was able to detect AIV and that the detection limit can reach 6.7 copies/μL and 12 copies/μL for the M and NP gene, respectively. In addition, this assay showed no cross-reactivity with other avian-derived RNA viruses such as NDV, IBDV, and IBV. Moreover, the detection system presented 97.5% consistency and agreement with rRT-PCR and virus isolation for detecting samples from poultry. This portable and accurate method has great potential for AIV detection in the field. Conclusion An RT-RPA/CRISPR method was developed for rapid, sensitive detection of AIV. The new system presents a good potential as an accurate, user-friendly, and inexpensive platform for point-of-care testing applications.