Newcastle Disease Virus Vaccine Research Articles

Recombinant fusion protein expression of Indonesian isolate Newcastle disease virus in Escherichia coli BL21(DE3)

Abstract. Wulanjati MP, Witasari LD, Wijayanti N, Haryanto A. 2021. Recombinant fusion protein expression of Indonesian isolate Newcastle disease virus in Escherichia coli BL21(DE3). Biodiversitas 22: 3249-3255. Newcastle disease is a major problem in poultry industry due to high mortality of susceptible chicken. New vaccine agents are important to be developed to eliminate the disease threat. This study aimed to examine the expression of fusion (F) protein of Newcastle Disease Virus (NDV) from Indonesian isolates in E. coli BL21(DE3) by IPTG induction. The sample was a part of F gene of NDV from Galur, Kulon Progo, Yogyakarta, Indonesia (0663/04/2013) with a molecular size of 600 bp that was synthesized and inserted into pBT7-N-His expression vector. The recombinant F protein with molecular weight of 25.6 kDa was successfully expressed in E. coli BL21(DE3), purified using Ni-NTA magnetic silica beads, and confirmed by western blotting. Optimization of expression showed that recombinant F protein was optimally expressed by induction of 1.0 mM IPTG when the cells reached OD600 = 0.6. The induction duration was 8 h. B-cell epitopes prediction showed that F protein possessed four epitopes that possibly recognized by B-cell. Since recombinant F protein was considered to possess immunogenicity, its potency as a candidate of NDV vaccine agent should be investigated in the future.

Comparison of the protective antigen variabilities of prevalent Newcastle disease viruses in response to homologous/heterologous genotype vaccines

The genotype VII Newcastle disease virus (NDV) vaccine has begun to replace the traditional genotype II NDV vaccine and is widely used in the commercial poultry of China. However, the effect of homologous and heterogeneous anti-NDV serum on the evolution of prevalent NDV is unknown. To understand the effect of genotype II and VII anti-NDV serum on the evolution of genotype VII NDV strains, ZJ1 (waterfowl origin) and CH/SD/2008/128 (ND128; chicken origin) were used for serial passage of 30 generations in DF-1 cells without anti-NDV serum or with genotype II and VII anti-NDV serum independently. The F and HN genes of the 2 viruses were amplified for the 10th, 20th, and 30th generations of each serial passage group and compared with their respective original viruses. We found that there was only one mutation at position 248 in the F gene of ZJ1 due to the serum pressure of genotype VII anti-NDV. Similarly, mutations at residue 527 of the F gene, and position 9 and 319 of the HN gene of ND128 were noted in both anti-NDV serum groups. The results show that the nonsynonymous (NS)-to-synonymous (S) ratio of the F gene of ZJ1 virus was 1.6, and for the HN gene, it was 2.5 in the anti-II serum group. In the anti-VII serum group, the NS/S ratio for the F gene was 2.1, and for the HN gene, it was 2.5. The NS/S ratio of the F gene of the ND128 virus was 0.8, and for the HN gene, it was 3 in the anti-II serum group. Furthermore, the NS/S ratio of the F gene was 0.8, and the HN gene was 2.3 in the anti-VII group. Taken together, our findings highlight that there was no significant difference in the variation of protective antigens in genotype VII NDV under the selection pressure of homologous and heterogeneous genotype NDV inactivated vaccines.

Thymic transcriptome analysis after Newcastle disease virus inoculation in chickens and the influence of host small RNAs on NDV replication

Newcastle disease (ND), caused by virulent Newcastle disease virus (NDV), is a contagious viral disease affecting various birds and poultry worldwide. In this project, differentially expressed (DE) circRNAs, miRNAs and mRNAs were identified by high-throughput RNA sequencing (RNA-Seq) in chicken thymus at 24, 48, 72 or 96 h post LaSota NDV vaccine injection versus pre-inoculation group. The vital terms or pathways enriched by vaccine-influenced genes were tested through KEGG and GO analysis. DE genes implicated in innate immunity were preliminarily screened out through GO, InnateDB and Reactome Pathway databases. The interaction networks of DE innate immune genes were established by STRING website. Considering the high expression of gga-miR-6631-5p across all the four time points, DE circRNAs or mRNAs with the possibility to bind to gga-miR-6631-5p were screened out. Among DE genes that had the probability to interact with gga-miR-6631-5p, 7 genes were found to be related to innate immunity. Furthermore, gga-miR-6631-5p promoted LaSota NDV replication by targeting insulin induced gene 1 (INSIG1) in DF-1 chicken fibroblast cells. Taken together, our data provided the comprehensive information about molecular responses to NDV LaSota vaccine in Chinese Partridge Shank Chickens and elucidated the vital roles of gga-miR-6631-5p/INSIG1 axis in LaSota NDV replication.

Pekin duck productivity, physiological stress, immune response and behavior under 20L:4D and 16L:8D photoperiods

As duck production becomes more prevalent on a global scale, modern duck housing systems must utilize artificial lighting photoperiods to maximize production and welfare; however, limited research has examined the manipulation of photoperiods on Pekin ducks. The objective of this study was therefore to determine the effects of two photoperiods – 20 h of light (L): 4 h of darkness (D) and 16 L:8D - on the growth, stress, and fear responses of 384 Pekin ducks during the grow-out period. The 20L:4D ducks had a more efficient feed conversion ratio (P < 0.02). The 20L:4D ducks were less stressed, as indicated by reduced plasma corticosterone concentration, heterophil to lymphocyte ratio, and composite asymmetry score (P < 0.02), and elevated humoral immune response to a Newcastle Disease Virus vaccine (P = 0.035) compared to 16L:8D ducks. These results indicated ducks reared under the 20L:4D photoperiod had more efficient nutrient metabolism and stronger humoral immune response to a Newcastle Disease Virus vaccine due to improved FCR and decreased stress and the effects of stress compared to the 16L:8D photoperiod group. Fear response data from this study was considered inconclusive, and future investigation is still required to elucidate how various photoperiod schedules influence the fear response of Pekin ducks. The results of this study emphasize the need for appropriate, species-specific artificial photoperiods in modern grow-out facilities to maximize the production of Pekin ducks while also reducing stress.

Newcastle disease vaccine virus I-2 fails to acquire virulence during repeated passage in vivo

Background This study determined whether the naturally attenuated, thermotolerant Newcastle disease vaccine virus I-2 could acquire virulence after five in vivo passages through SPF chickens. Methods Study design was to international requirements including European Pharmacopoeia, Ph. Eur., v9.0 04/2013:0450, 2013. I-2 Working Seed (WS) was compared with five-times-passaged I-2 WS (5XP WS) in intracerebral pathogenicity index (ICPI), Fo cleavage site sequencing and Safety tests. Results The first passage series used a 50% brain: 50% tracheal tissue challenge homogenate and was unsuccessful as I-2 was not detected after the fourth passage. A second passage series used 10% brain: 90% tracheal tissue homogenates. I-2 was isolated from tracheal tissue in each passage. However harvested titres were below the minimum challenge level (107 EID50) specified for the ICPI and Safety tests, possibly reflecting I-2’s inherently low pathogenicity (interestingly caecal tonsils yielded significant titres). Given this the WS and 5XP WS comparisons proceeded. ICPI values were 0.104 and 0.073 for the WS group and the 5XP WS group respectively confirming that I-2, whether passaged or not, expressed low pathogenicity. F0 amino-acid sequences for both WS and 5XP WS were identified as 112R-K-Q-G-R-↓-L-I-G119 and so compatible with those of avirulent ND viruses. In safety, no abnormal clinical signs were observed in both groups except for two chicks in the 5XP WS group, where one bird was withdrawn due to a vent prolapse, and another bird died with inconclusive necropsy results. Conclusions: These data, the issue of low passage titres with little or no virus isolation from brain tissues and the genomic copy approach suggest a need to amend Ph. Eur. v9.0 04/2013:0450, 2013 for naturally attenuated, low pathogenicity vaccine viruses such as I-2. These results add to the literature and field data demonstrating that Newcastle Disease vaccine virus I-2 is safe for use.

Impact of routine Newcastle disease vaccination on chicken flock size in smallholder farms in western Kenya.

Poultry represent a widely held economic, nutritional, and sociocultural asset in rural communities worldwide. In a recent longitudinal study in western Kenya, the reported mean number of chickens per household was 10, with increases in flock size constrained principally by mortality. Newcastle disease virus is a major cause of chicken mortality globally and hypothesized to be responsible for a large part of mortality in smallholder flocks. Our goal was to determine the impact of routine Newcastle disease virus (NDV) vaccination on flock size and use this data to guide programs to improve small flock productivity. We conducted a factorial randomized controlled trial in 537 households: in 254 households all chickens were vaccinated every 3 months with I-2 NDV vaccine while chickens in 283 households served as unvaccinated controls. In both arms of the trial, all chickens were treated with endo- and ecto parasiticides every 3 months. Data on household chicken numbers and reported gains and losses were collected monthly for 18 months. Consistent with prior studies, the overall flock size was small but with increases in both arms of the study over time. The mean number of chickens owned at monthly census was 13.06±0.29 in the vaccinated households versus 12.06±0.20 in the control households (p = 0.0026) with significant gains in number of chicks (p = 0.06), growers (p = 0.09), and adults (p = 0.03) in the vaccinated flocks versus the controls. Household reported gains were 4.50±0.12 total chickens per month when vaccinated versus 4.15±0.11 in the non-vaccinated controls (p = 0.03). Gains were balanced by voluntary decreases, reflecting household decision-making for sales or household consumption, which were marginally higher, but not statistically significant, in vaccinated households and by involuntary losses, including mortality and loss due to predation, which were marginally higher in control households. Quarterly NDV vaccination and parasiticidal treatment resulted in an increase in flock size by a mean of one bird per household as compared to households where the flock received only parasiticidal treatment. While results suggest that the preventable fraction of mortality attributable to Newcastle disease is comparatively small relatively to all-cause mortality in smallholder households, there was a significant benefit to vaccination in terms of flock size. Comparison with previous flock sizes in the study households indicate a more significant benefit from the combined vaccination and parasiticidal treatment, supporting a comprehensive approach to improving flock health and improving household benefits of production in the smallholder setting.

Effects of heat stress on the immune responses of chickens subjected to thermal manipulation in the pre-hatch period

Heat stress affects the performance of poultry species and also induces immunosuppression. Chickens can be treated by thermal conditioning to have better heat stress tolerance. Our purpose was to determine the effect of acute heat stress on the immune response, i.e. antibody production against Newcastle disease virus (NDV) and change in the proportion of leukocyte components, in chicks subjected to prenatal heat conditioning. Eighty as-hatched broiler chicks from the same parent stock were used: control (40 chicks incubated at 36.7 °C from days 18-20 of embryonic life) and thermally manipulated (TM) (40 chicks incubated at 38.4 °C from day 18-20 of embryonic life; 4 h/day). The chickens were exposed to heat stress: at day 19 (31 °C/8 h) and at day 35 (32 °C/10 h). The first heat stress (day 19) decreased the lymphocyte counts and significantly increased the heterophil counts (P < 0.05) in both treatments (from 34.25 to 55% in the controls and from 37 to 60.06% in the TM chicks). The second heat stress (day 35) did not alter the heterophil and lymphocyte profiles of the chickens. Before the heat stress, all of the chickens (control and TM) presented the same positive antibody titres to NDV vaccination. After the first heat stress, 50% of the control samples and 40% of samples from the TM chickens were negative. After the second heat stress, 80% of the TM samples were negative.

Protection against Different Genotypes of Newcastle Disease Viruses (NDV) Afforded by an Adenovirus-Vectored Fusion Protein and Live NDV Vaccines in Chickens

The efficacy of an adenovirus-vectored Newcastle disease virus (NDV) vaccine expressing the fusion (F) NDV protein (adeno-F) was evaluated against challenges with virulent heterologous and homologous NDV strains to the F protein. In a preliminary study, two different doses (low and high) of adeno-F were tested against a virulent NDV strain containing the homologous NDV F protein, CA02. In a second study, at three weeks post-vaccination, the efficacy of the high dose of adeno-F was compared to a live attenuated NDV vaccine strain (LaSota) against three antigenically distinct virulent NDV challenge strains, one homologous (CA02) and two heterologous (TZ12, EG14) to F in the vectored vaccine. In both experiments, clinical signs, mortality, virus shedding, and humoral response were evaluated. In the first experiment, the survival rates from birds vaccinated with adeno-F at a high and low dose were 100% and 25%, respectively. In the second experiment, birds vaccinated with the high dose of adeno-F had a survival rate of 80%, 75%, and 65% after challenge with the CA02, TZ12, and EG14 viruses, respectively. All of the LaSota-vaccinated birds survived post-challenge no matter the NDV challenge strain. High antibody titers were detected after vaccination with LaSota by HI and ELISA tests. The majority of adeno-F-vaccinated birds had detectable antibodies using the ELISA test, but not using the HI test, before the challenge. The data show that both the similarity of the F protein of the adeno-F vaccine to the challenge virus and the adeno-F vaccination dose affect the efficacy of an adenovirus-vectored NDV vaccine against a virulent NDV challenge.

Identification of a potential neutralizing linear epitope of hemagglutinin-neuraminidase in Newcastle disease virus

BackgroundThe hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) is a major antigen that can induce protective antibodies in poultry. However, its antigenic epitopes have not been fully elucidated. Therefore, defining the linear epitopes of HN, especially neutralizing epitopes, will be useful for revealing its antigenic characterization.MethodsIn this study, we analyzed B-cell immunodominant epitopes (IDEs) of the HN protein from the vaccine strain LaSota using pepscan technology with LaSota-specific chicken hyperimmune antisera. We constructed IDEs-RFP plasmids and prepared anti-IDEs peptide mouse sera to identify IDEs through immunological tests. At last, the different diluted anti-IDE antisera were used in BHK-21 cells to perform the neutralization test.ResultsFive IDEs of the HN were screened and further verified by indirect immunofluorescence assays, dot blots and Western blots with NDV- and IDEs-specific antisera. All five IDEs showed good immunogenicity. IDE5 (328–342 aa) could recognize only class II NDV but did not react with the class I strain. Most of the IDEs are highly conserved among the different strains. A neutralization test in vitro showed that the peptide-specific mouse antisera of IDE4 (242–256 aa) and HN341-355, a reported neutralizing linear epitope, could partially neutralize avirulent LaSota as well as virulent strains at similar levels, suggesting that IDE4 might be a potential neutralizing linear epitope.ConclusionThe HN protein is a major protective antigen of NDV that can induce neutralizing antibodies in animals. We identified five IDEs of the HN using a pepscan approach with NDV-specific chicken hyperimmune antisera. The five IDEs could elicit specific antibodies in mice. IDE4 (242–256 aa) was identified as a novel potential neutralizing linear epitope. These results will help elucidate the antigenic epitopes of the HN and facilitate the development of NDV vaccines.

Molecular mechanism of enhancing the immune effect of the Newcastle disease virus vaccine in broilers fed with Bacillus cereus PAS38.

The aim of this study was to explore the molecular mechanism of enhancing the immune effect of the Newcastle disease virus (NDV) vaccine in broilers fed with Bacillus cereus PAS38. The results showed that the NDV antibody titer of broilers in the treatment group supplemented with B. cereus PAS38 was higher than that of the control group, and the difference was significant at 28 days of age (P < 0.05). The spleen, thymus and bursa of fabricius of 42-day-old broilers were quickly collected to construct a differentially expressed gene library of suppression subtractive hybridization (SSH). A total of 31 immune-related differentially expressed genes were screened from three immune organs, of which 15 were up-regulated and 16 were down-regulated. After silencing the up-regulated genes MIF, CD74, DOCK2 and KLHL6, the expression levels of cytokines (Akirin2, NF-κB, IL-2, IL-4, IL-6, IFN-γ and TNF-α) in lymphocytes were reduced to varying degrees. B. cereus PAS38 might be involved in the proliferation, differentiation, activation, migration of B lymphocytes and vaccine antigen presentation by up-regulating the expression of MIF, CD74, DOCK2, KLHL6 and other genes. Moreover, it also stimulated plasma cells to produce immunoglobulins and specific antibodies, thereby improving the humoral immune function of broilers and enhancing the immune effect of the NDV vaccine.

Effect of a probiotic (

Context Deoxynivalenol (DON) contamination of feedstuffs causes detrimental effects on animals and poultry. Dietary inclusion of microbial feed additives, such as probiotics and/or yeast, seems to be a useful approach for DON detoxification and reducing the toxin absorption from the gut. Aims This study was conducted to evaluate the synergetic effects of a probiotic (Lactobacillus spp.), yeast (Saccharomyces cerevisiae) and mycotoxin detoxifier on performance, serum chemical parameters and immune status of broiler chickens fed a DON-contaminated diet. Methods A total of 200 1-day-old female broilers (Hubbard®) were allocated to five dietary treatments with four replicates each in a completely randomised design. Experimental diets consisted of: (1) control diet (basal diet), (2) DON diet (basal diet contaminated with 10 mg/kg DON), (3) DON diet supplemented with 0.25% mycotoxin detoxifier (Mycofix® Plus), (4) DON diet supplemented with a combination of 0.4 g/kg probiotic and 1.5 g/kg yeast, and (5) DON diet supplemented with a combination of a probiotic, yeast and mycotoxin detoxifier. Key results The results showed that the DON diet significantly increased the feed conversion ratio compared with mycotoxin detoxifier and control diets. Administration of 0.4 g/kg probiotic along with 1.5 g/kg yeast to a DON-contaminated diet caused a significant enhancement of dressing percentage, leg relative weight and aspartate aminotransferase enzyme activity. The DON-contaminated diet reduced total protein, albumin, triglyceride and cholesterol concentrations, but consumption of a probiotic, yeast and mycotoxin detoxifier improved the levels of these parameters. The DON-treated birds showed a reduction in haematocrit, haemoglobin, red blood cells and blood lymphocyte percentage in comparison with control levels. The inclusion of a combination of a probiotic and yeast along with mycotoxin detoxifier diminished the increased heterophil: lymphocyte ratio by DON administration. The antibody titre against Newcastle disease virus vaccine increased by inclusion of a probiotic, yeast and Mycofix. Conclusions In summary, a combination of a probiotic and yeast along with mycotoxin detoxifier can improve the enzyme activity, immunity and haematological attributes, which are useful for DON-contaminated diet detoxification. Implications Dietary supplementation with a probiotic, yeast and mycotoxin detoxifier could be used as an alternative to detoxification DON in broiler chickens.

A rapid response vaccine development strategy for newcastle disease in poultry

Poultry is a major source of protein in sub-Saharan Africa and many other lower-income regions. Newcastle disease virus (NCDV) comprises a significant threat toward poultry production. While NCDV vaccines are routinely used in developed countries, those used in sub-Saharan Africa are mostly imported and are not specific to locally circulating strains. Indeed, the lack of rapid, field-based NCDV detection and the absence of cost-effective production methods for pure, strain-specific vaccines hampers efficient poultry production throughout these regions. This remains a major problem for both subsistence and commercial farming.The aim for this study was firstly, to develop a field-based isothermal PCR assay for NCDV detection that employed a portable instrument and real-time data transfer application. Secondly,to use the nucleic acid sequence data obtained from field isolates to develop a protocol compatible with rapid emergency vaccine production for NCDV.To achieve this, the isothermal PCR detection assay was applied to field isolates from suspected NCDV outbreaks on commercial poultry farms in KwaZulu-Natal, South Africa, while for the vaccine development, the NCDV matrix gene of one of the isolates was sequenced and used to design primers for the recombinant cloning of this antigen into an adenoviral vector.This‘vaccine vector’ and a control adenoviral vector were each amplified in 293T cells and then used to infect both 293T cells as a production cell line and chicken embryo fibroblasts (CEF) as a preliminary model of the target host. Western blotting confirmed the successful expression of the V5epitopetagby the control vector in both cell lines, which established the compatibility of the adenovirus vector as an appropriate carrier of the target antigen. Mass spectrometry confirmed expression of the NCDV matrix protein by the vaccine vector in both cell lines. In conclusion, the improved turnaround time from detection to the production of the vaccine antigen was under6weeks.The approach described here provides a rapid and cost-effective protocol for both the pathogen detection on-site and the production of pure vaccine antigens specific to an emerging field strain of NCDV within lower-income regions.

Different kinetics of chicken interferon-alpha signalling transduction responses following immunization of broiler chickens with different Newcastle disease virus vaccines and infection with virulent genotype VIId strain

ABSTRACT Newcastle disease virus (NDV) is a highly contagious and notifiable avian disease leading to grave economic losses in the poultry industry. Although the immune responses against NDV have been widely investigated, little is known regarding the virus interaction with the host innate immune responses. In this study, we tested the effect of different commercially applied Newcastle disease vaccines as well as virulent NDV genotype VIId on the expression pattern of the upstream regulator and downstream effector genes related to chicken interferon-alpha (chIFNα) signalling transduction pathway. Using quantitative real-time PCR analysis, mild transient induction of chIFNα-inducible genes was detected in bird spleen 72 h post-vaccination (hpv) with either live LaSota (respiratory) or VG/GA (enteric) strains. Vaccination with the enteric VG/GA strain led to stimulation of the investigated pathway as early as 24 hpv which continued up to 7 days in bird caecal tonsils. Subcutaneous injection with inactivated LaSota oil adjuvant-based vaccine led to continual stimulation of the investigated pathway up to 7 days post-vaccination (dpv). The recombinant herpesvirus of turkey (rHVT) - NDV vaccine led to remarkable stimulation of all the tested cytokines up to 17 dpv in comparison with LaSota and VG/GA NDV vaccines. Stronger but transient activation of all the tested cytokines was detected in spleens during the first 24 h post-challenge with virulent NDV (vNDV) which reduced gradually and diminished later due to the virus-induced lymphocytic depletion. This study will aid in the discovery of new approaches to control NDV.

DIPG-38. ADDITION OF MULTIMODAL IMMUNOTHERAPY TO COMBINATION TREATMENT STRATEGIES FOR CHILDREN WITH DIPG: FINAL RESULTS OF A COHORT OF CHILDREN

The prognosis of children with Diffuse Intrinsic Pontine Glioma (DIPG) remains dismal in spite of radio- and chemotherapy or therapies based on molecular biology diagnostics. Immunotherapy is a powerful and promising approach for improving the overall survival (OS). A retrospective analysis for feasibility, immune responsiveness and OS was performed on 41 children treated in compassionate use with Newcastle Disease Virus, hyperthermia and autologous dendritic cell vaccines as part of an individualized combined treatment approach for DIPG patients at diagnosis (n=28), or at time of progression (n=13). All except one patient had reduced values of at least one immune test before starting immunotherapy. In all patients at least one PanTum Detect test was outside the normal range. Ten patients had PDL1 mRNA expression in circulating tumor cells at diagnosis. Multimodal immunotherapy was feasible as scheduled, until progression, in all patients without major toxicity. When immunotherapy was part of primary treatment, median PFS and OS were 8.4m and 14.4m respectively with a 2-year OS of 10.7%. When immunotherapy was given at the time of progression, median PFS and OS calculated from diagnosis were 6.5m and 9.1m respectively. Th1 shift and rise in PanTum Detect test scores were linked with longer OS. Multimodal immunotherapy is feasible without major toxicity, and its value as part of a combination treatment for primary diagnosed DIPG should be elaborated in clinical trials.

Mulberry leaf polysaccharide supplementation contributes to enhancing the respiratory mucosal barrier immune response in Newcastle disease virus–vaccinated chicks

Despite high global vaccination coverage, Newcastle disease (ND) remains a constant threat to poultry producers owing to low antibody levels. Given the respiratory mucosa is the important site for Newcastle disease virus (NDV) vaccination, enhancing respiratory mucosal immunity may help control ND. Our previous study showed that mulberry leaf polysaccharide (MLP) is very promising in delivering a robust balanced immune response, but the effects of it on respiratory immunity in chicks are unknown. In this study, we evaluated the potential of MLP to activate respiratory mucosal immunity and revealed the possible mechanism of MLP as an immunopotentiator for ND vaccines. Chicks were randomly divided into 5 groups: blank control, vaccination control (VC), and low-, middle-, and high-dose MLP (MLP-L, MLP-M, and MLP-H) (n = 30). The serum results of humoral and cell-mediated immune responses showed significant increases in NDV hemagglutination inhibition antibody titer, IgG and IgA antibody levels, and the T-lymphocyte population in the MLP-M group compared with the VC group. Validation of results also indicated remarkable increases in tracheal antibody-mediated immunity and a mucosal immune response in the MLP-M group. Furthermore, the upregulation of TLR7 revealed a possible mechanism. Our findings provided evidence to consider MLP as a potential mucosal vaccine adjuvant candidate against ND in chickens.

Newcastle disease virus (NDV) expressing the spike protein of SARS-CoV-2 as a live virus vaccine candidate.

BackgroundDue to the lack of protective immunity of humans towards the newly emerged SARS-CoV-2, this virus has caused a massive pandemic across the world resulting in hundreds of thousands of deaths. Thus, a vaccine is urgently needed to contain the spread of the virus.MethodsHere, we describe Newcastle disease virus (NDV) vector vaccines expressing the spike protein of SARS-CoV-2 in its wild type format or a membrane-anchored format lacking the polybasic cleavage site. All described NDV vector vaccines grow to high titers in embryonated chicken eggs. In a proof of principle mouse study, the immunogenicity and protective efficacy of these NDV-based vaccines were investigated.FindingsWe report that the NDV vector vaccines elicit high levels of antibodies that are neutralizing when the vaccine is given intramuscularly in mice. Importantly, these COVID-19 vaccine candidates protect mice from a mouse-adapted SARS-CoV-2 challenge with no detectable viral titer and viral antigen in the lungs.InterpretationThe results suggested that the NDV vector expressing either the wild type S or membrane-anchored S without the polybasic cleavage site could be used as live vector vaccine against SARS-CoV-2.FundingThis work is supported by an NIAID funded Center of Excellence for Influenza Research and Surveillance (CEIRS) contract, the Collaborative Influenza Vaccine Innovation Centers (CIVIC) contract, philanthropic donations and NIH grants.

Yeast cell wall product enhanced intestinal IgA response and changed cecum microflora species after oral vaccination in chickens

The study was designed to explore the effect of a commercial yeast cell wall product (YP) on chicken intestinal IgA response and cecum microbiome after oral vaccination. Chickens were fed with YP during the experiments and orally immunized with live Newcastle disease virus (NDV) vaccine at 2 wk of age. Then, the animals were sacrificed, and samples were collected to measure the indicators of hemagglutination inhibition (HI), IgA response, IgA + cells, and cecum microbiome populations. The results showed that supplement of YP significantly enhanced serum NDV HI titer, intestinal NDV-specific secretory IgA, and intestinal IgA + cells. The sequencing results revealed that obviously increased relative abundance of Ruminococcaceae and decreased population of Bacteroidaceae in cecum were found in YP group. In summary, YP supplementation in diet enhanced intestinal IgA response to NDV vaccination by oral route and modulated the cecum microbiota to the advantage of the host in chickens.

Effect of diets containing earthworm powder and vermihumus on egg production, hatchability, blood parameters and immunity of Japanese breeder quails

The present study is aimed to evaluate the effects of different levels of earthworm (EW) powder and vermihumus (VH) on the growth performance, humoral immunity response, plasma constituents and reproductive performance of Japanese breeder quails. A total of 336 birds were assigned to seven treatments in a completely randomized design from day 30 to 90 of age. The dietary treatments were carried out, one without any addition of EW and VH, the second had a standard rate of VH at 0.8%, while a range of EW additions was supplied from 0%, 0.5%, 1%, 1.5%, 2% and 2.5%. The effects of treatments on body weight and mortality were not significant (p>.05). The highest feed intake was observed in birds fed the diets supplemented with 1% and 1.5% EW (p<.05). The highest titre of antibodies against influenza virus and sheep red blood cells belonged to the EW and VH treatments (p<.05). The antibody titre against Newcastle disease virus vaccine was not affected by the treatments. In general, the birds receiving EW and VH had the highest egg weight, egg production and egg mass weight (p<.05). Nonetheless, various levels of EW powder negatively affected hatchability and increased mortality and leg disorders (p<.05). The results showed that the levels of dietary 1% and 1.5% of EW powder gave the best growth and reproductive performance to birds respectively.

Protective efficacy of inactivated Newcastle disease virus vaccines prepared in two different oil-based adjuvants

Despite the availability of Newcastle disease (ND) vaccines for more than six decades, disease outbreaks continue to occur with huge economic consequences to the global poultry industry. The aim of this study is to develop a safe and effective inactivated vaccine based on a recently isolated Newcastle disease virus (NDV) strain IBS025/13 and evaluate its protective efficacy in chicken following challenge with a highly virulent genotype VII isolate. Firstly, high titre of IBS025/13 was exposed to various concentrations of binary ethylenimine (BEI) to determine the optimal conditions for complete inactivation of the virus. The inactivated virus was then prepared in form of a stable water-in-oil emulsion of black seed oil (BSO) or Freund’s incomplete adjuvant (FIA) and used as vaccines in specific pathogen-free chicken. Efficacy of various vaccine preparations was also evaluated based on the ability of the vaccine to protect against clinical disease, mortality and virus shedding following challenge with highly virulent genotype\\VII NDV isolate. The results indicate that exposure of NDV IBS025/13 to 10 mM of BEI for 21 h at 37 °C could completely inactivate the virus without tempering with the structural integrity of the viral hemagglutin-neuraminidase protein. More so, the inactivated vaccines adjuvanted with either BSO- or FIA-induced high hemagglutination inhibition antibody titre that protected the vaccinated birds against clinical disease and in some cases virus shedding, especially when used together with live attenuated vaccines. Thus, genotype VII-based NDV-inactivated vaccines formulated in BSO could substantially improve poultry disease control particularly when combined with live attenuated vaccines.

Residues 315 and 369 in HN Protein Contribute to the Thermostability of Newcastle Disease Virus.

Thermostable Newcastle disease virus (NDV) vaccines have been widely used in areas where a “cold-chain” is not reliable. However, the molecular mechanism of NDV thermostability remains poorly understood. In this work, we constructed chimeric viruses by exchanging viral fusion (F) and/or hemagglutinin-neuraminidase (HN) genes between the heat-resistant strain HR09 and thermolabile strain La Sota utilizing a reverse genetic system. The results showed that only chimeras with HN derived from the thermostable virus exhibited a thermostable phenotype at 56°C. The hemagglutinin (HA) and neuraminidase (NA) activities of chimeras with HN derived from the HR09 strain were more thermostable than those containing HN from the La Sota strain. Then, we used molecular dynamics simulation at different temperatures (310 K and 330 K) to measure the HN protein of the La Sota strain. The conformation of an amino acid region (residues 315–375) was observed to fluctuate. Sequence alignment of the HN protein revealed that residues 315, 329, and 369 in the La Sota strain and thermostable strains differed. Whether the three amino acid substitutions affected viral thermostability was investigated. Three mutant viruses based on the thermolabile strain were generated by substituting one, two or three amino acids at positions 315, 369, and 329 in the HN protein. In comparison with the parental virus, the mutant viruses containing mutations S315P and I369V possessed higher thermostablity and HA titers, NA and fusion activities. Taken together, these data indicate that the HN gene of NDV is a major determinant of thermostability, and residues 315 and 369 have important effects on viral thermostability.