Raw white rice has not been considered a good carrierfor oral vaccination, probably because of its antiviralactivity. Methods are required to overcome antiviralactivity in raw white rice. This study was carried out todetermine the effects of various treatments of raw whiterice on the survival of strain I-2 of Newcastle diseasevirus. These included cooking and baking the rice ormixing the rice with vegetable oil prior to coating withvaccine virus. The vaccine-coated rice was then stored for30min and 24h, followed by quantitative recovery of thevirus. Thirty min after mixing, uncooked, cooked, andbaked rice, and rice mixed with vegetable oil showed titersof 10(6.2), 10(7.2), 10(6.6), and 10(7.0) EID50/0.1ml, respectively.After storage for 24h at 22-25oC, the titers dropped to10(5.0), 10(6.5), 10(5.0), and 10(6.0) EID50/0.1ml for uncooked,cooked, baked, and oiled rice, respectively.
Animals ; Chick Embryo ; Chickens ; Cookery ; Newcastle Disease/*virology ; Newcastle disease virus/growth & development/*physiology ; Oryza sativa/chemistry/*virology ; Viral Vaccines/chemistry

Newcastle disease viruses (NDVs) cause systemic diseases in chickens with high mortality. However, little is known about persistence of NDVs in contaminated tissues from infected birds. In this study, we examined viral replication in the feather pulp of chickens inoculated with viscerotropic velogenic NDV (vvNDV) genotype VII. Reverse transcription real-time PCR and immunohistochemistry were used to investigate viral persistence in the samples. vvNDV was detected in the oropharynx and cloaca and viral antigens were detected in the feathers, suggesting that feathers act as sources of viral transmission.
Animals ; Antigens, Viral/analysis ; Chickens ; Cloaca/virology ; Feathers/*virology ; Microbial Viability ; Newcastle Disease/transmission/*virology ; Newcastle disease virus/isolation & purification/*physiology ; Oropharynx/virology ; Poultry Diseases/transmission/*virology ; Virus Replication/*physiology

Elution time of velogenic, mesogenic and lentogenic strains of Newcastle disease virus was determined. The differences in their elution time were also calculated. Four samples, each of a velogenic strain (VGF2), a mesogenic strain (Komarov) and a lentogenic strain (LaSota) were used for hemagglutination test with 0.6% chicken red blood cells. The time it took for wells of the end hemagglutination points (highest dilution that gave agglutination) to elute was recorded as elution time for each sample. The mean elution time of the three strains of Newcastle disease virus differed significantly (p < 0.05). The velogenic strain gave the highest mean elution time of 118 min, followed by the mesogenic strain with 59 min and the lentogenic strain with 25 min. Based on this result it appears that elution time could form a basis for rough characterization of isolates of Newcastle disease virus into the three major strains.
Animals ; Chickens/blood ; Erythrocytes/*virology ; Hemagglutination Tests ; *Hemagglutination, Viral ; Newcastle disease virus/isolation&purification/*pathogenicity

Based on the genomic sequence of NDV08-004 strain (GenBank accession number FJ794269), seven pairs of primers were designed to amplify the genomic fragments by RT-PCR and cloned into pGEM-Teasy vector. The fragments (named A to G) were sub-cloned into transcription vector pOLTV5 according to the universal RE site and the plasmid named NDV08-004-pO which contained the full length cDNA of NDV08-004 strain was constructed. Three helper plasmids (pCI-NP, pCI-P and pCI-L) together with NDV08-004-pO were co-transfected into BSR T7/5 cells, and the transfection supernatant was inoculated into SPF embryonated eggs to rescue the virus. The virus was rescued successfully and identified by HA and RT-PCR and sequencing. The rescue system constructed in this study provided a good foundation for the further related research.
Animals ; Base Sequence ; Chick Embryo ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Newcastle Disease ; virology ; Newcastle disease virus ; genetics ; Plasmids ; Reverse Genetics ; methods

OBJECTIVETo explore changes in structure and function of the mitochondria of human gastric carcinoma BGC-823 cells after Newcastle disease virus (NDV) infection.

METHODSElectron microscopy was applied to observe the structure of mitochondria; Rhodamine 123 staining was used to determine the mitochondrial membrane potential; the activity of Na(+)-K(+)-ATPase and Ca(2+)-ATPase were also determined and the release of cytochrome C was detected by Western blotting.

RESULTSThe structure of mitochondria in the tumor cells infected with NDV changed distinctly. In the infected group the activity of mitochondrial Na(+)-K(+)-ATPase and Ca(2+)-ATPase significantly declined (P < 0.01), and compared with control cells, mitochondrial trans-membrane potential was decreased. NDV infection induced the decrease of cytochrome C levels.

CONCLUSIONThe effects of NDV infection on the structure and functions of mitochondria of human gastric carcinoma BGC-823 cells might play a role in the oncolysis of NDV.

Animals ; Carcinoma ; enzymology ; metabolism ; virology ; Cell Line, Tumor ; Chick Embryo ; Cytochromes c ; metabolism ; Humans ; Membrane Potential, Mitochondrial ; Mitochondria ; enzymology ; metabolism ; virology ; Newcastle Disease ; enzymology ; metabolism ; virology ; Newcastle disease virus ; physiology ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Stomach Neoplasms ; enzymology ; metabolism ; virology

Two hundred thirty specimens of wild birds were collected from some areas in Heilongjiang Province during the period of 2003~2004, including two batches of specimens collected randomly from a same flock of mallards in Zhalong Natural Reserve in August and December, 2004, respectively. Primary virus isolation and identification for avian influenza virus (AIV) and Newcastle disease virus (NDV) were performed. The results showed that only two specimens of young mallards collected from Zhalong Natural Reserve in August, 2004 were positive to AIV (isolation rate 0.9%), and one strain (D57) of these two virus isolates was identified to be H9 subtype by hemagglutination inhibition test. Meanwhile, the two batches of blood serum samples of mallards from Zhalong were also examined for antibodies against AIV and NDV. Among 38 blood serum samples collected in August, antibodies against the hemagglutinin of H1, H3, H5, H6 and H9 subtypes of AIV were found in 1, 0, 2, 0 and 8 samples, respectively; and 11 samples were found with antibody against NDV. Whereas the NDV isolation in both two batches of specimens of mallard was negative, all of the 32 blood serum samples collected in December were negative for antibodies against AIV and NDV.
Animals ; Animals, Wild/*virology ; Antibodies, Viral/isolation&purification ; Birds/virology ; China/epidemiology ; Hemagglutination Tests ; Influenza A virus/*isolation&purification ; Influenza in Birds/epidemiology/immunology/*virology ; Newcastle Disease/epidemiology/immunology/*virology ; Newcastle disease virus/*isolation&purification ; Reverse Transcriptase Polymerase Chain Reaction

In order to determine the challenge dose of pigeon paramyxovirus type 1 (PPMV-1) inactivated vaccine (S-1 strain). The virus titer of PPMV-1 E5 allantoic fluid (Chuansha strain) was determined using SPF chicken embryos in this research. After inoculating 30-day-old and 120-day-old pigeons with low-HI antibody against PPMV-1 (HI antibody < or =2) with different doses of PPMV-1 (Chuansha strain), the clinical symptoms and histopathological lesions of the challenged pigeons were examined. The results showed that the minimal lethal dose (MLD) of PPMV-1 (Chuansha strain) was 102.5 ELD50, so we determined that 10(5.5) ELD50, which was 1000 times the MLD, could be taken as the challenge dose in the vaccine efficacy test for PPMV-1 inactivated vaccine (S-1 strain).
Animals ; Antibodies, Viral ; immunology ; Bird Diseases ; immunology ; mortality ; virology ; Chick Embryo ; Columbidae ; immunology ; virology ; Newcastle Disease ; immunology ; mortality ; virology ; Newcastle disease virus ; immunology ; pathogenicity ; Phylogeny ; Viral Vaccines ; immunology ; Virulence

Small interfering ribonucleic acid (siRNA)-induced RNA degradation can inhibit viral infection, and has been investigated extensively for its efficacy as antiviral therapy. The potential therapeutic role of lentiviral-mediated short hairpin ribonucleic acid (shRNA) to Newcastle disease virus (NDV) replication in vivo has been explored less often. We constructed two recombinant lentiviral vectors containing shRNA against the phosphoprotein (P) of the NDV, RNAi-341 and RNAi-671. Recombinant shRNA lentivirus vectors were co-transfected into 293T cells, along with helper plasmids, to package the recombinant shRNA lentivirus. Lentivirus-based shRNAs were titrated and transduced into NDV-susceptible chicken embryo fibroblasts (CEFs) and chick embryos. Antiviral activity against the NDV strain was evaluated by virus titration and real-time reverse transcription-polymerase chain reaction. RNAi-341 and RNAi-671 strongly suppressed transient expression of a FLAG-tagged P fusion protein in 293T cells. RNAi-341 and RNAi-671 NDV reduced virus titers by 66.6-fold and 30.6-fold, respectively, in CEFs 16 h after infection. RNAi-341 and RNAi-671 reduced virus titers in specific pathogen-free chick embryos by 99% and 98%, respectively, 48 h after infection. Both shRNAs inhibited accumulation of not only P-gene mRNA, but also nucleocapsid, M-, F-, HN-, and L-gene mRNA. RNAi-341 silenced P-gene mRNA more potently than RNAi-671. These results suggest that shRNAs silencing the P gene had substantial antiviral properties and inhibited NDV replication in CEFs and chick embryos.
Animals ; Chick Embryo ; Chickens ; Down-Regulation ; Fibroblasts ; virology ; Gene Targeting ; Lentivirus ; genetics ; metabolism ; Newcastle Disease ; virology ; Newcastle disease virus ; genetics ; physiology ; Phosphoproteins ; genetics ; metabolism ; Poultry Diseases ; virology ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism ; Virus Replication

An outbreak of fever and dyspnea with high incidence rate and case fatality rate occurred among pigs in a pigfarm in Jilin province, China, in 2000. The paramyxovirus-like particles could be observed in lungs, spleens and kidneys of the dead pigs under the transmission electron microscope. A Newcastle disease virus (NDV) isolate designated as JL01 was determined as the causal agent for the disease outbreak. The purified virus was reinoculated into pigs, and then the pigs infected with the virus showed similar symptoms, and the HI antibody of NDV could be detected from the reinoculated pigs. The MDT, ICPI and EID50 of the JL01 isolate was 55.2h, 1.60 and 10(-7.5)/0.1 mL respectively. The F gene of JL01 was cloned and sequenced, and the results showed that the identities of F gene shared by JL01 and avirulent NDV reference strains were from 91.5% to 98.5%, and could be ascribed to NDV genotype I. Thus, the swine NDV JL01 strain should be an avirulent strain with gene variation, but the virulence of JL01 was just the same as velogenic strains.
Animals ; Chickens ; Dogs ; Horses ; Mice ; Microscopy, Electron ; Newcastle Disease ; virology ; Newcastle disease virus ; classification ; genetics ; isolation & purification ; ultrastructure ; Phylogeny ; Polymerase Chain Reaction ; Rabbits ; Sequence Analysis, DNA ; Swine ; Swine Diseases ; virology ; Viral Proteins ; genetics

Three Newcastle disease virus (NDV) strains recovered from ND outbreaks in chickens and duck flocks in north china during 2009 to 2011 were completely sequenced and biologically characterized. All the strains were velogenic and had the velogenic motif 112R-R-Q-K-R-F117 which was consistent with the results of biological tests. Analysis of the variable region (nucleotide 47 to 420) of the F gene indicated that the three isolates belonged to genotype VII d. Cross hemagglutination inhibition test indicated that the antigen homology between three isolates and LaSota were 82.5%-89.4%, the homology between the two isolates from chicken was 90%. A cross-protection experiment in which specific-pathogen-free chickens vaccinated with LaSota were challenged by SDLY01 isolate showed that LaSota vaccine could provide complete protection against SDLY01, however virus discharge could be detected on fifth day. Challenge experiment in which Cherry Valley duck of 30 day old challenged with SD03 strain indicated that cherry valley duck had no disease in experiment period, but virus discharge could be detected from Larynx and cloaca until fifth day. Genome length of three NDV isolates was 15192bp and belonged to genotype VII d. Sequence analysis clarified that the whole genomic sequence of these three isolates shared high homology with NDV virus strains isolated from goose and duck over the same period, which elucidated that NDV isolated from goose, duck or chicken had close genetics and epidemiological relationship.
Amino Acid Sequence ; Animals ; Bird Diseases ; virology ; Chickens ; Columbidae ; Ducks ; Geese ; Genome, Viral ; Molecular Sequence Data ; Newcastle Disease ; virology ; Newcastle disease virus ; chemistry ; classification ; genetics ; isolation & purification ; Phylogeny ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics