Newcastle Disease Virus Manipulates Mitochondrial MTHFD2-Mediated Nucleotide Metabolism for Virus Replication.

Viruses require host cell metabolic reprogramming to satisfy their replication demands; however, the mechanism by which the Newcastle disease virus (NDV) remodels nucleotide metabolism to support self-replication remains unknown. In this study, we demonstrate that NDV relies on the oxidative pentose phosphate pathway (oxPPP) and the folate-mediated one-carbon metabolic pathway to support replication. In concert with [1,2-13C2] glucose metabolic flow, NDV used oxPPP to promote pentose phosphate synthesis and to increase antioxidant NADPH production. Metabolic flux experiments using [2,3,3-2H] serine revealed that NDV increased one-carbon (1C) unit synthesis flux through the mitochondrial 1C pathway. Interestingly, methylenetetrahydrofolate dehydrogenase (MTHFD2) was upregulated as a compensatory mechanism for insufficient serine availability. Unexpectedly, direct knockdown of enzymes in the one-carbon metabolic pathway, except for cytosolic MTHFD1, significantly inhibited NDV replication. Specific complementation rescue experiments on small interfering RNA (siRNA)-mediated knockdown further revealed that only a knockdown of MTHFD2 strongly restrained NDV replication and was rescued by formate and extracellular nucleotides. These findings indicated that NDV replication relies on MTHFD2 to maintain nucleotide availability. Notably, nuclear MTHFD2 expression was increased during NDV infection and could represent a pathway by which NDV steals nucleotides from the nucleus. Collectively, these data reveal that NDV replication is regulated by the c-Myc-mediated 1C metabolic pathway and that the mechanism of nucleotide synthesis for viral replication is regulated by MTHFD2. IMPORTANCE Newcastle disease virus (NDV) is a dominant vector for vaccine and gene therapy that accommodates foreign genes well but can only infect mammalian cells that have undergone cancerous transformation. Understanding the remodeling of nucleotide metabolic pathways in host cells by NDV proliferation provides a new perspective for the precise use of NDV as a vector or in antiviral research. In this study, we demonstrated that NDV replication is strictly dependent on pathways involved in redox homeostasis in the nucleotide synthesis pathway, including the oxPPP and the mitochondrial one-carbon pathway. Further investigation revealed the potential involvement of NDV replication-dependent nucleotide availability in promoting MTHFD2 nuclear localization. Our findings highlight the differential dependence of NDV on enzymes for one-carbon metabolism, and the unique mechanism of action of MTHFD2 in viral replication, thereby providing a novel target for antiviral or oncolytic virus therapy.

Dietary inclusion of mushroom (Flammulina velutipes) stem waste on growth performance and immune responses in growing layer hens.

BACKGROUND: Medicinal mushrooms contain biologically active substances that can be used as an immune-modulating agent in poultry. The present study aimed to investigate the effects of Flammulina velutipes mushroom waste (FVW) on performance, immune response and serum immunity in growing layer hens. RESULTS: No significant differences (P > 0.05) were observed with respect to average daily feed intake, body weight gain and feed conversion ratio among the experimental groups during the entire study period (1-70 days). Antibody titers against Newcastle disease and infectious bronchitis were higher (P < 0.05) in the FVW fed groups than in the control and antibiotic groups. On day 28, serum immunoglobulin (Ig)A and IgG were higher (P < 0.05) in the 6% FVW group than in the control and antibiotic fed groups. On day 70, serum IgA was higher (P < 0.05) in FVW fed groups than in the control group; IgG was higher (P < 0.05) in the FVW groups than in the control and antibiotic groups. However, IgM was higher (P < 0.05) in both the 4% and 6% FVW groups than in the control and antibiotic groups for both experimental periods. Serum cytokine interleukin (IL)-2 and tumor necrosis factor-α concentrations were significantly higher (P < 0.05) in both the 4% and 6% FVW grousp than in the control and antibiotic groups; IL-4 was significantly higher (P < 0.05) in the FVW groups than in the control group; and IL-6 was significantly higher (P < 0.05) in the 6% FVW group than in the control and antibiotic groups. CONCLUSION: FVW at the 6% level can be used as a potential phytogenic feed stuff in growing layer hen rations with respect to improving the immune response without affecting normal weight gain. © 2018 Society of Chemical Industry.

Genomic and biological characterization of Newcastle disease viruses isolated from migratory mallards (Anas platyrhynchos).

Given the global evolutionary dynamics of Newcastle disease viruses (NDVs), it is imperative to continue extensive surveillance, routine monitoring and characterization of isolates originating from natural reservoirs (waterfowls). In this report, we isolated and characterized two virulent NDV strains from clinically healthy mallard (Anas platyrhynchos). Both isolates had a genome of 15,192 nucleotides encoding six genes in an order of 3´-NP-P-M-F-HN-L-5´. The biological characteristics (mean death time: 49.5-50 hr, EID50108.5 ml-1) and presence of a typical cleavage site in the fusion (F) protein (112R-R-Q-K-R↓F117) confirmed the velogenic nature of these isolates. Phylogenetic analysis classified both isolates as members of genotype VII within class-II. Furthermore, based upon the hypervariable region of the F gene (375 nt), isolates showed clustering within sub-genotype VIIi. Similarity index and parallel comparison revealed a higher nucleotide divergence from commonly used vaccine strains; LaSota (21%) and Mukteswar (17%). A comparative residues analysis with representative strains of different genotypes, including vaccine strains, revealed a number of substitutions at important structural and functional domains within the F and hemagglutinin-neuraminidase (HN) proteins. Together, the results highlight consistent evolution among circulating NDVs supporting extensive surveillance of the virus in waterfowl to better elucidate epidemiology, evolutionary relationships and their impacts on commercial and backyard poultry.

Newcastle disease virus-like particles induce dendritic cell maturation and enhance viral-specific immune response.

Circulating of genotype VII Newcastle disease virus (NDV) is a great threat to the poultry industry worldwide. Virus-like particles (VLPs) are increasingly being considered as potential viral vaccines due to their safety and efficacy. In this study, we analyzed in vitro the stimulatory effects of VLPs containing the matrix and hemagglutinin-neuraminidase of genotype VII NDV on dendritic cells (DCs) and evaluated their immunogenicity in mice. The results showed that immature bone marrow-derived dendritic cells (BMDCs) responded to stimulation with VLPs by up-regulating expressions of MHC II, CD40, CD80, and CD86 molecules and by increasing the cytokine secretions of TNF-α, IFN-γ, IL-6, and IL-12p70. Besides, VLPs enhanced the immunostimulatory capacity of DCs to stimulate autologous T cell proliferation. Furthermore, VLPs can induce efficient humoral and cellular immune responses, and recruit mature DCs to the spleen in C57BL/6 mice, as shown by an obvious increase in double-positive proliferation of splenic CD11c+CD86+ cells. These data indicate that NDV VLPs can be a valuable candidate for NDV vaccine development.

Virus interference between H7N2 low pathogenic avian influenza virus and lentogenic Newcastle disease virus in experimental co-infections in chickens and turkeys.

Low pathogenicity avian influenza virus (LPAIV) and lentogenic Newcastle disease virus (lNDV) are commonly reported causes of respiratory disease in poultry worldwide with similar clinical and pathobiological presentation. Co-infections do occur but are not easily detected, and the impact of co-infections on pathobiology is unknown. In this study chickens and turkeys were infected with a lNDV vaccine strain (LaSota) and a H7N2 LPAIV (A/turkey/VA/SEP-67/2002) simultaneously or sequentially three days apart. No clinical signs were observed in chickens co-infected with the lNDV and LPAIV or in chickens infected with the viruses individually. However, the pattern of virus shed was different with co-infected chickens, which excreted lower titers of lNDV and LPAIV at 2 and 3 days post inoculation (dpi) and higher titers at subsequent time points. All turkeys inoculated with the LPAIV, whether or not they were exposed to lNDV, presented mild clinical signs. Co-infection effects were more pronounced in turkeys than in chickens with reduction in the number of birds shedding virus and in virus titers, especially when LPAIV was followed by lNDV. In conclusion, co-infection of chickens or turkeys with lNDV and LPAIV affected the replication dynamics of these viruses but did not affect clinical signs. The effect on virus replication was different depending on the species and on the time of infection. These results suggest that infection with a heterologous virus may result in temporary competition for cell receptors or competent cells for replication, most likely interferon-mediated, which decreases with time.

Strong innate immune response and cell death in chicken splenocytes infected with genotype VIId Newcastle disease virus.

BACKGROUND: Genotype VIId Newcastle disease virus (NDV) isolates induce more severe damage to lymphoid tissues, especially to the spleen, when compared to virulent viruses of other genotypes. However, the biological basis of the unusual pathological changes remains largely unknown. METHODS: Virus replication, cytokine gene expression profile and cell death response in chicken splenocytes infected with two genotype VIId NDV strains (JS5/05 and JS3/05), genotype IX NDV strain F48E8 and genotype IV NDV strain Herts/33 were evaluated. Statistical significance of differences between experimental groups was determined using the Independent-Samples T test. RESULTS: JS5/05 and JS3/05 caused hyperinduction of type I interferons (IFNs) (IFN-α and -ß) during detection period compared to F48E8 and Herts/33. JS5/05 increased expression level of IFN-γ gene at 6 h post-inoculation (pi) and JS3/05 initiated sustained activation of IFN-γ within 24 h pi, whereas transcriptional levels of IFN-γ remained unchanged at any of the time points during infection of F48E8 and Herts/33. In addition, compared to F48E8 and Herts/33, JS3/05 and JS5/05 significantly increased the amount of free nucleosomal DNA in splenocytes at 6 and 24 h pi respectively. Annexin-V and Proidium iodid (PI) double staining of infected cells showed that cell death induced by JS3/05 and JS5/05 was characterized by marked necrosis compared to F48E8 and Herts/33 at 24 h pi. These results indicate that genotype VIId NDV strains JS3/05 and JS5/05 elicited stronger innate immune and cell death responses in chicken splenocytes than F48E8 and Herts/33. JS5/05 replicated at a significantly higher efficiency in splenocytes than F48E8 and Herts/33. Early excessive cell death induced by JS3/05 infection partially impaired virus replication. CONCLUSIONS: Viral dysregulaiton of host response may be relevant to the severe pathological manifestation in the spleen following genotype VIId NDV infection.

Perturbations in the antioxidant metabolism during Newcastle disease virus (NDV) infection in chicken : protective role of vitamin E.

The aim of the present study was to investigate the effect of vitamin E on pro/anti-oxidant status in the liver, brain and heart of Newcastle disease virus (NDV) infected chickens. Activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione-S-transferase (GST) and the levels of reduced glutathione and malonaldehyde were estimated in selected tissues of uninfected, NDV-infected and NDV + vit. E-treated chickens. A significant increase in MDA levels in brain and liver (p < 0.05) was observed in NDV-infected chickens when compared to controls. The activities of SOD, CAT, GPx, GR, GST and levels of GSH were significantly (p < 0.05) decreased in brain and liver of NDV-infected chickens over controls. On the other hand, a significant decreased MDA levels and enhanced antioxidant enzyme activity levels were observed in NDV + vit. E-treated animals compared to NDV-infected chickens. Histopathological studies revealed that liver of NDV infected chicken shows focal coagulation and infiltration of hepatocytes, whereas neuronal necrosis and degeneration of Purkinje cells were observed in brain and moderate infiltration of inflammatory cells was observed in heart. However such histological alterations were not observed in NDV + vit. E-treated animals. The results of the present study, thus demonstrated that antioxidant defense mechanism is impaired after the induction of NDV, suggesting its critical role in cellular injury in brain and liver. Further, the results also suggest that vitamin E treatment will ameliorate the antioxidant status in the infected animals. The findings could be beneficial to understand the role of oxidative stress in the pathogenesis of NDV and therapeutic interventions of antioxidants.

Combined exposure of Japanese quails to cyanotoxins, Newcastle virus and lead: oxidative stress responses.

Wild birds are continually exposed to many anthropogenic and natural stressors in their habitats. Over the last decades, mass mortalities of wild birds constitute a serious problem and may possibly have more causations such as natural toxins including cyanotoxins, parasitic diseases, industrial chemicals and other anthropogenic contaminants. This study brings new knowledge on the effects of controlled exposure to multiple stressors in birds. The aim was to test the hypothesis that influence of cyanobacterial biomass, lead and antigenic load may combine to enhance the effects on birds, including modulation of antioxidative and detoxification responses. Eight treatment groups of model species Japanese quail (Coturnix coturnix japonica) were exposed to various combinations of these stressors. The parameters of detoxification and oxidative stress were studied in liver and heart after 30 days of exposure. The antioxidative enzymatic defense in birds seems to be activated quite efficiently, which was documented by the elevated levels and activities of antioxidative and detoxification compounds and by the low incidence of damage to lipid membranes. The greatest modulations of glutathione level and activities of glutathione-S-transferase, glutathione peroxidase, glutathione reductase, superoxide dismutase, catalase and lipid peroxidation were shown mostly in the groups with combined multiple exposures. The results indicate that the antioxidative system plays an important role in the protective response of the tissues to applied stressors and that its greater induction helps to protect the birds from more serious damage. Most significant changes of these "defense" parameters in case of multiple stressors suggest activation of this universal mechanism in situation with complex exposure and its crucial role in protection of the bird health in the environment.

Infection dynamics of highly pathogenic avian influenza and virulent avian paramyxovirus type 1 viruses in chickens, turkeys and ducks.

A range of virus doses were used to infect 3-week-old chickens, turkeys and ducks intranasally/intraocularly, and infection was confirmed by the detection of virus shedding from the buccal or cloacal route by analysis of swabs collected using real-time reverse transcriptase-polymerase chain reaction assays. The median infectious dose (ID(50)) and the median lethal dose (LD(50)) values for two highly pathogenic avian influenza (HPAI) viruses of H5N1 and H7N1 subtypes and one virulent Newcastle disease virus (NDV) were determined for each virus and host combination. For both HPAI viruses, turkeys were >100-fold more susceptible to infection than chickens, while both these hosts were >10-fold more susceptible to H5N1 virus than the H7N1 virus. All infected chickens and turkeys died. Ducks were also much more readily infected with the H5N1 virus (ID(50)< or =10(1) median embryo infective dose [EID(50)]) than the H7N1 virus (ID(50)=10(4.2) EID(50)). However, the most notable difference between the two viruses was their virulence for ducks, with a LD(50) of 10(3) EID(50) for the H5N1 virus, but no deaths in ducks being attributed to infection with H7N1 virus even at the highest dose (10(6) EID(50)). For both HPAI virus infections of ducks, the ID(50) was lower than the LD(50), indicating that infected birds were able to survive and thus excrete virus over a longer period than chickens and turkeys. The NDV strain used did not appear to establish infection in ducks even at the highest dose used (10(6) EID(50)). Some turkeys challenged with 10(6) EID(50), but not other doses, of NDV excreted virus for a number of days (ID(50)=10(4.6) EID(50)), but none died. In marked contrast, chickens were shown to be extremely susceptible to infection and all infected chickens died (ID(50)/LD(50)=10(1.9) EID(50)).

Newcastle disease virus-induced cytopathic effect in infected cells is caused by apoptosis.

The velogenic Newcastle disease virus (NDV) causes highly infectious and economically significant Newcastle disease (ND) in birds of various species. In cell culture NDV induces cytopathic effect (CPE) characterized by rounding, vacuolation, syncytia formation and cell death. Aside from cell to cell fusion caused by the F and HN glycoprotein of the virus molecular events leading to cell death are not known. In the current study, NDV-infected Vero cells, at 48 h p.i., showed nuclear condensation, cytoplasm blebbing, DNA fragmentation, and phosphatidylserine translocation to the cell surface. In addition, virus-infected cells demonstrated decreased DNA content and an increased Bax to Bcl-2 ratio, p53 level and caspase 3, 8, 9 expression compared to mock-infected cells. Based on these results, it was concluded that CPE in NDV-infected cells was caused by to the induction of apoptosis with the involvement of p53 and the Bax, dependent apoptotic pathways.

MicroRNA gga-miR-455-5p suppresses Newcastle disease virus replication via targeting cellular suppressors of cytokine signaling 3.

Newcastle disease (ND) is an acute and contagious avian disease caused by Newcastle disease virus (NDV). MicroRNAs (miRNAs) play a significant role in host-pathogen interactions and the innate immune response. However, the role of miRNAs in the host response to NDV infection is not clearly understood. In this study, we showed that expression of the cellular miRNA gga-miR-455-5p was downregulated in vivo and in vitro in response to NDV infection. Next, we found that the transfection of chicken embryonic fibroblasts (CEFs) with gga-miR-455-5p suppressed NDV replication, while the blockade of endogenous gga-miR-455-5p expression with inhibitors enhanced NDV replication. In addition, gga-miR-455-5p enhanced the expression of type I interferon and the interferon-inducible genes (ISGs) OASL and Mx1 by targeting SOCS3, a negative regulator of type I IFN signaling. Altogether, these findings highlight the crucial role of gga-miR-455-5p in host defense against NDV by targeting the SOCS3 gene to inhibit NDV replication.

Haemagglutination inhibition antibodies, rectal temperature and total protein of chickens infected with a local Nigerian isolate of velogenic Newcastle disease virus.

Changes in values of haemagglutination inhibition (HI) antibody titre, rectal temperature (RT) and total protein (TP) were determined for Shaver Brown chickens infected with Newcastle disease virus (NDV) Kudu 113. The infected chickens came down with Newcastle disease by day 3 post infection (PI). The major clinical signs were depression, greenish diarrhoea, paralysis of legs and wings, opisthotonus and torticolis. Mortality and morbidity were 52% and 1000%, respectively. There were haemorrhagic lesions in the wall of the intestine, proventricular mucosa and caecal tonsils. There were necrosis and mononuclear cell infiltration of the liver, kidney and spleen. There was a significant increase in daily mean HI antibody titres from days 3 to 9 PI. Similarly, significant rise in daily mean RTs were noticed in the infected chickens from days 1 to 13 PI. On the other hand, there was a decrease in daily mean TP concentrations of infected chickens, beginning from day 3 PI, and the lowest concentration of 2.60 +/- 0.15 g/dl was obtained by days 7 and 11 PI. The values of HI, RT and TP for the control chickens were relatively constant during the experiment. The correlation coefficient (r) between HI and RT was positive and highly significant (r = 0.725, p<0.001), while the relationship between HI and TP was negative but highly significant (r = -0.712, p<0.001). It was concluded that NDV Kudu 113 induced increases in values of HI and RT, which occurred concurrently with a decrease in TP concentrations of infected chickens.

Neuroendocrine peptide hormones and their receptors in the immune system. Production, processing and action.

Recent findings indicate that the immune and neuroendocrine systems interact and modulate one another functionally. The mechanism for this seems to be that the 2 systems share a set of receptors and ligands (hormones). Cells of the immune system are able to synthesize neuroendocrine peptide hormones which are biologically active and produced in physiologically significant quantities. Furthermore, leukocytes possess functional receptors for these same neuroendocrine hormones which will specifically modulate immune responses. The structural and functional evidence for these interactions is reviewed and discussed in the context of a bidirectional regulatory circuit between the immune and neuroendocrine systems.

Neurochemical and neuroendocrine responses to Newcastle disease virus administration in mice.

Mice injected intraperitoneally with Newcastle disease virus (NDV) responded with increased plasma concentrations of ACTH and corticosterone and increased hypothalamic concentrations of the tryptophan and of the norepinephrine catabolite, 3-methoxy,4-hydroxyphenylethyleneglycol (MHPG) and the serotonin catabolite, 5-hydroxyindoleacetic acid (5-HIAA). Two different strains of NDV, a lentogenic and a mesogenic one, elicited dose-dependent effects in these responses. Both strains elicited near maximal responses at doses around 1000 hemagglutination units. The maximal effects on ACTH, corticosterone and MHPG occurred around 2 h, but the effects on tryptophan and 5-HIAA were greatest at 8 h. Similar responses in plasma corticosterone, and cerebral tryptophan and 5-HIAA were observed following i.p. injection of polyinosinic-polycytidylic acid, but MHPG was not altered. The cyclo-oxygenase inhibitor, indomethacin, had little effect on the NDV-induced increases in plasma corticosterone and ACTH, and hypothalamic indolamines, but essentially ablated the MHPG response. The effect of NDV on plasma corticosterone, like that of endotoxin (LPS), was prevented by hypophysectomy, suggesting that the pituitary was required for these responses. These endocrine and neurochemical responses to NDV resemble those to interleukin-1 (IL-1) and LPS. Therefore we tested mice pretreated with the IL-1-receptor antagonist. This treatment prevented the neurochemical and plasma ACTH and corticosterone responses to IL-1, but did not alter those to LPS, and prevented the endocrine and neurochemical responses to NDV in approximately half of the animals. Thus IL-1 may be a mediator of the responses to NDV, but additional factors may also be involved.

Effects of Newcastle disease virus infection on the binding, phagocytic, and bactericidal activities of respiratory macrophages of the turkey.

Effects of Newcastle disease virus (NDV) infection on the binding, phagocytic, and bactericidal activities of turkey respiratory macrophages were studied. Respiratory macrophages of the turkey demonstrated the presence of immunoglobulin (Ig) G and complement receptors but lacked IgM receptors. Respiratory macrophages from NDV-infected turkeys showed little or no depression of binding of sheep erythrocyte-IgG complexes and sheep erythrocyte-IgM-complement complexes to their appropriate membrane receptors. In contrast, respiratory macrophages from NDV-infected turkeys showed significant (P less than or equal to 0.05) depression of phagocytosis of similar complexes. Bacterial killing by respiratory macrophages from NDV-infected turkeys was significantly (P less than or equal to 0.05) inhibited.

Effect of social stress on Newcastle disease virus (LaSota) infection.

Individual immature chickens were socially stressed when placed into an established group of chickens. The stress response was determined by periodic measurement of plasma cortisol and by determination of the relative regression of the bursa of Fabricius. Stressed chickens had elevated cortisol levels during the first 24 hours following the initiation of social stress. The bursa-to-body-weight ratios of stressed birds at 96 hours following stress were significantly lower than the ratios in control birds, and their tuberculin reaction was significantly depressed. Newcastle disease virus was more invasive and attained higher titers in stressed birds. The hemagglutination-inhibition (HI) response of the stressed birds was higher in one trial and lower in another.

Efficacy of viable and inactivated Newcastle disease virus vaccines in turkeys.

Market turkeys spray-vaccinated at 20 days of age with viable Newcastle disease virus (NDV) vaccine and challenged 7 weeks postvaccination failed to yield NDV by tracheal swabbing 4 days postchallenge but demonstrated serologic evidence of infection. Birds vaccinated subcutaneously with inactivated oil-emulsion (OE) NDV vaccine had virologic and serologic evidence of infection. Breeder hens vaccinated by spray with commercial La Sota vaccine at 19 weeks of age and revaccinated subcutaneously with OE vaccine at 32 weeks of age had an adequate level of resistance against a drop in egg production but demonstrated serologic evidence of infection when challenged with velogenic NDV at 38 weeks of age.

Tracheal mucus transport rate and bacterial clearance in turkeys exposed by aerosol to La Sota strain of Newcastle disease virus.

Tracheal mucus transport rate (TMTR) and quantitative clearance of aerosolized Escherichia coli from the trachea, lung, and air sac were measured in healthy unanesthetized turkeys and in turkeys exposed by aerosol to a La Sota vaccine strain of Newcastle disease virus (NDV). The TMTR of uninfected turkeys was 42.4 +/- 14.7 cm/min. The TMTR of NDV-infected turkeys was depressed on days 3 through 7 postexposure (PE); depression was significant (P less than or equal to 0.05) on day 7 PE. Tracheal E. coli clearance in NDV-infected turkeys was reduced on days 4 through 9 PE, significantly so on day 5 PE (P less than or equal to 0.01). Depression of TMTR and tracheal E. coli clearance were associated histologically with replacement of normal pseudostratified columnar epithelium by 3 to 8 layers of immature nonciliated cells. E. coli clearance by the lung and air sac of NDV-infected turkeys was depressed on days 5 through 9 PE.

Experimental infection in turkeys and chickens with Ornithobacterium rhinotracheale.

Ornithobacterium rhinotracheale was found to cause growth retardation in both turkeys and chickens after experimental intra-air sac administration and to cause growth retardation together with airsacculitis and pneumonia after aerosol administration. Both turkey and chicken isolates of O. rhinotracheale were able to induce the same kind of respiratory inflammations and weight-gain losses in chickens as well as turkeys. Turkey rhinotracheitis virus was found to have a triggering effect on the O. rhinotracheale infection in turkeys, and Newcastle disease virus and to a lesser extent infectious bronchitis virus showed triggering effects on the O. rhinotracheale infection in chickens. Ornithobacterium rhinotracheale could be reisolated from affected organs of experimentally infected birds.

Influence of Newcastle disease virus on the severity of Pasteurella anatipestifer infection in turkeys.

This study was designed to examine whether vaccine or virulent strains of Newcastle disease virus (NDV) would potentiate the disease caused by Pasteurella anatipestifer infection in turkeys. The studies were conducted in turkeys of two age groups. There were three experiments. In two experiments four-week-old turkeys were exposed either to vaccine or virulent strains of NDV after experimental P anatipestifer infection. In the third experiment 14-week-old turkeys were first exposed to virulent NDV and superimposed with P anatipestifer infection. In experiment 1, one bird died where P anatipestifer was given in combination with the vaccine strain of NDV. However, there was no difference in the clinical signs, gross lesions and histopathology compared with turkeys given P anatipestifer alone. In experiment 3 where turkeys received a virulent strain of NDV in combination with P anatipestifer, birds became dyspnoeic and showed signs of illness. There was a difference in the course of the disease, gross lesions and histopathology when compared with turkeys that received P anatipestifer only.