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2.
Poult Sci ; 99(4): 1928-1938, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32241473

RESUMO

In this study, we isolated and identified 2 infectious bronchitis virus (IBV) strains from layer chickens soon after vaccination with the Massachusetts-Connecticut bivalent vaccine (Conn) and H120 and 4/91 booster vaccines in China in 2011. The results of cross-virus-neutralization tests and phylogenetic analysis of the S1 subunit of spike gene of these vaccine strains and other reference strains showed that strain LJL/110302 was of GI-19 lineage, whereas LLN/111169 was of the GI-1 lineage of the Conn serotype. Further comparative genomic analysis revealed that LLN/111169, an IBV strain with novel traits, originated from multiple recombination events (at least 3 recombination sites) between GI-19 and the Conn and 4/91 vaccine strains. LLN/111169 was pathogenic to specific pathogen-free (SPF) chickens. This is of prime importance because while IBV prevention measures worldwide are mainly dependent on modified live vaccine strains, our results showed that recombination between field and vaccine strains has produced a novel pathogenic IBV strain. In addition, LLN/111169 showed relatively broad tissue tropism (trachea, lungs, kidneys, and cecal tonsils) in infected SPF chickens. These results emphasize the importance of IBV surveillance in chicken flocks.


Assuntos
Galinhas , Infecções por Coronavirus/veterinária , Vírus da Bronquite Infecciosa/fisiologia , Vírus da Bronquite Infecciosa/patogenicidade , Doenças das Aves Domésticas/virologia , Replicação Viral , Animais , Antígenos Virais/análise , China , Infecções por Coronavirus/virologia , Vírus da Bronquite Infecciosa/genética , Recombinação Genética , Estudos Retrospectivos , Sorogrupo , Organismos Livres de Patógenos Específicos , Vacinas Atenuadas/análise , Vacinas Virais/análise , Virulência
4.
J Appl Microbiol ; 128(1): 65-73, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31562676

RESUMO

AIMS: To compare antigen extraction efficiency of chemical methods such as benzyl alcohol, chloroform, sodium citrate, extraction buffer with Tween-20 (EBT) and isopropyl myristate for determination of 146S content in the fresh and stored FMD oil-adjuvanted vaccines. METHODS AND RESULTS: Standard vaccine with antigen payload of 10, 5 and 5 µg per cattle dose (2 ml) for serotypes O, A and Asia1, respectively, was used to compare the antigen extraction efficiency of five chemical methods: benzyl alcohol, chloroform, sodium citrate, EBT buffer and isopropyl myristate. The purity of the extracted 146S antigen was quantified by caesium chloride (CsCl) ultracentrifugation. Serotype-specific sandwich ELISA (sELISA) was developed to identify the serotype and to compare the 146S in aqueous phase and ultrafractions. The antigen recovery was also tested in stored trivalent vaccine. Coefficient of regression was calculated to assess the predictive power of the benzyl alcohol extraction method. Of the five methods, benzyl alcohol showed consistent antigen recovery of >90% in monovalent as well as trivalent vaccines. Ultrafraction showed a 1·4 ratio at A259/239 nm in UV spectrophotometry indicating the presence of 146S. sELISA revealed that the antigen recovery was significantly less in ultrafractions than that of aqueous phase. Further, there was no significant difference in antigen recovery from stored trivalent vaccine for 12 months, indicating the usefulness of the benzyl alcohol method. Linear regression model revealed R2  = 0·99 with a narrow band of predictive interval. CONCLUSIONS: The benzyl alcohol method was efficient in extracting 146S from the monovalent and trivalent fresh and stored FMD vaccines. CsCl density gradient precisely quantified the 146S, while sELISA identified the serotype of the vaccine. SIGNIFICANCE AND IMPACT OF THE STUDY: When the benzyl alcohol method is coupled with CsCl density gradient and sELISA, it has the potential to determine the 146S content of FMD vaccine.


Assuntos
Antígenos Virais/isolamento & purificação , Vírus da Febre Aftosa/imunologia , Febre Aftosa/virologia , Vacinas Virais/imunologia , Adjuvantes Imunológicos , Animais , Antígenos Virais/imunologia , Bovinos , Ensaio de Imunoadsorção Enzimática , Vírus da Febre Aftosa/genética , Sorogrupo , Potência de Vacina , Vacinas Virais/análise
5.
Fish Shellfish Immunol ; 96: 223-234, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31821845

RESUMO

In the past decades, the aquaculture industry made great progress in China, which contributes more than 70% yield of the world's farmed fish. Along with the rapid growth of fish production, increased emergence and outbreak of numbers of diseases pose harm to the aquaculture industry and food safety. From the efficient, safe, environmental and ethical aspects, vaccines is definitely the most appropriate and focused method to control different kinds of fish diseases. In China, researchers have done huge works on the fish vaccines, and so far six domestic aquatic vaccine products along with one imported aquatic vaccine have obtained the national veterinary medicine certificate. More critically, some new vaccines have also entered the field experiment stage and showed broad market prospects. In the present review, authors summarize seven aquatic vaccines, including the live vaccine against grass carp hemorrhagic disease, the inactivated vaccine against Aeromonas hydrophila sepsis in fish, the live vaccine against Edwardsiella tarda in turbot, the anti-idiotypic antibody vaccine against Vibrio alginolyticus, V. parahaemolyticus, and E. tarda in Japanese flounder, the cell-cultured inactivated vaccine against grass carp hemorrhagic disease, the inactivated vaccine against fish infectious spleen and kidney necrosis virus (ISKNV), and the genetically engineered live vaccine against V. anguillarum in turbot. Moreover, different delivery routes of fish vaccines are also compared in this review, along with differential fish immune response after vaccination. All these efforts will ultimately benefit the healthy and sustainable development of aquaculture industry in China.


Assuntos
Vacinas Bacterianas/uso terapêutico , Doenças dos Peixes/prevenção & controle , Vacinas Virais/uso terapêutico , Animais , Vacinas Bacterianas/análise , China , Vacinas Virais/análise
6.
J Med Entomol ; 56(6): 1463-1466, 2019 10 28.
Artigo em Inglês | MEDLINE | ID: mdl-31549715

RESUMO

West Nile virus (WNV) (Flaviviridae: Flavivirus) was discovered in Africa more than 80 yr ago and became recognized as an avian pathogen and a cause of neurologic disease in horses largely during periodic incursions into Europe. Introduction of WNV into North America stimulated great anxiety, particularly in the equine industry, but also for pet owners and livestock producers concerned about the effect of WNV on other domestic animals. Numerous subsequent studies of naturally occurring and experimentally induced disease greatly expanded our understanding of the host range and clinical consequences of WNV infection in diverse species and led to rapid development and deployment of efficacious vaccines for horses. In addition to humans, horses are clearly the animals most frequently affected by serious, sometimes lethal disease following infection with WNV, but are dead-end hosts due to the low-magnitude viremia they develop. Dogs, cats, and livestock species including chickens are readily infected with WNV, but only occasionally develop clinical disease and are considered dead-end hosts for the virus.


Assuntos
Vacinação/veterinária , Vacinas Virais , Febre do Nilo Ocidental/veterinária , Vírus do Nilo Ocidental , Vacinas Virais/administração & dosagem , Vacinas Virais/análise , Febre do Nilo Ocidental/prevenção & controle
7.
Biomed Res Int ; 2019: 2750472, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31223613

RESUMO

Chicken infectious anemia virus (CIAV) causes the atrophy of bone marrow hematopoietic and lymphoid tissues in chicks, leading to huge economic losses all over the world. The using of attenuated vaccine contaminated with CIAV increased the mortality and the pathogenicity of other diseases in many farms. However, it is difficult to detect the CIAV contamination by general detection technology due to the extremely low dose of CIAV in vaccines. In this study, we established a new method called droplet digital Polymerase Chain Reaction (ddPCR) to detect CIAV contamination of vaccines more sensitively and accurately. The lowest detection limitation of this method is 2.4 copies of CIAV plasmid or CIAV contamination at 0.1 EID50/1000 feathers in vaccines without any positive signals of other viruses. Besides, the sensitivity of ddPCR is 100 times greater than that of conventional PCR and 10 times greater than that of real-time PCR. The ddPCR technique is more sensitive and more intuitive. Therefore, it could be valuable for the detection of CIAV contamination in vaccines.


Assuntos
Vírus da Anemia da Galinha/genética , Galinhas/virologia , Contaminação de Medicamentos , Reação em Cadeia da Polimerase , Vacinas Virais/análise , Animais , Vacinas Atenuadas/análise , Vacinas Virais/genética
8.
J Biomed Sci ; 26(1): 47, 2019 Jun 19.
Artigo em Inglês | MEDLINE | ID: mdl-31215493

RESUMO

Non-polio enteroviruses are emerging viruses known to cause outbreaks of polio-like infections in different parts of the world with several cases already reported in Asia Pacific, Europe and in United States of America. These outbreaks normally result in overstretching of health facilities as well as death in children under the age of five. Most of these infections are usually self-limiting except for the neurological complications associated with human enterovirus A 71 (EV-A71). The infection dynamics of these viruses have not been fully understood, with most inferences made from previous studies conducted with poliovirus.Non-poliovirus enteroviral infections are responsible for major outbreaks of hand, foot and mouth disease (HFMD) often associated with neurological complications and severe respiratory diseases. The myriad of disease presentations observed so far in children calls for an urgent need to fully elucidate the replication processes of these viruses. There are concerted efforts from different research groups to fully map out the role of human host factors in the replication cycle of these viral infections. Understanding the interaction between viral proteins and human host factors will unravel important insights on the lifecycle of this groups of viruses.This review provides the latest update on the interplay between human host factors/processes and non-polio enteroviruses (NPEV). We focus on the interactions involved in viral attachment, entry, internalization, uncoating, replication, virion assembly and eventual egress of the NPEV from the infected cells. We emphasize on the virus- human host interplay and highlight existing knowledge gaps that needs further studies. Understanding the NPEV-human host factors interactions will be key in the design and development of vaccines as well as antivirals against enteroviral infections. Dissecting the role of human host factors during NPEV infection cycle will provide a clear picture of how NPEVs usurp the human cellular processes to establish an efficient infection. This will be a boost to the drug and vaccine development against enteroviruses which will be key in control and eventual elimination of the viral infections.


Assuntos
Infecções por Enterovirus/virologia , Enterovirus/fisiologia , Fatores Hospedeiros de Integração/fisiologia , Vacinas Virais/análise , Vírion/fisiologia , Humanos
9.
Electrophoresis ; 40(18-19): 2277-2284, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30951206

RESUMO

A CZE method was validated and implemented for fast and accurate in-process determination of adenovirus concentrations of downstream process samples obtained during manufacturing of adenovirus vector-based vaccines. An analytical-quality-by-design approach was embraced for method development, method implementation, and method maintenance. CZE provided separation of adenovirus particles from sample matrix components, such as cell debris, residual DNA and proteins. The intermediate precision of the virus particle concentration was 6.9% RSD and the relative bias was 2.3%. In comparison, the CZE method is intended to replace a quantitative polymerase chain reaction method which requires three replicates in three analytical runs to achieve an intermediate precision of 8.1% RSD. Given that, in addition, the time from sampling till reporting results of the CZE method was less than 2 h, whereas quantitative polymerase chain reaction requires 3 days, it follows that the CZE method enables faster processing times in downstream processing.


Assuntos
Adenoviridae , Eletroforese Capilar/métodos , Vírion , Adenoviridae/química , Adenoviridae/isolamento & purificação , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Projetos de Pesquisa , Vacinas Virais/análise , Vacinas Virais/química , Vírion/química , Vírion/isolamento & purificação , Cultura de Vírus
10.
Vet Q ; 39(1): 26-55, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31006350

RESUMO

Nipah (Nee-pa) viral disease is a zoonotic infection caused by Nipah virus (NiV), a paramyxovirus belonging to the genus Henipavirus of the family Paramyxoviridae. It is a biosafety level-4 pathogen, which is transmitted by specific types of fruit bats, mainly Pteropus spp. which are natural reservoir host. The disease was reported for the first time from the Kampung Sungai Nipah village of Malaysia in 1998. Human-to-human transmission also occurs. Outbreaks have been reported also from other countries in South and Southeast Asia. Phylogenetic analysis affirmed the circulation of two major clades of NiV as based on currently available complete N and G gene sequences. NiV isolates from Malaysia and Cambodia clustered together in NiV-MY clade, whereas isolates from Bangladesh and India clusterered within NiV-BD clade. NiV isolates from Thailand harboured mixed population of sequences. In humans, the virus is responsible for causing rapidly progressing severe illness which might be characterized by severe respiratory illness and/or deadly encephalitis. In pigs below six months of age, respiratory illness along with nervous symptoms may develop. Different types of enzyme-linked immunosorbent assays along with molecular methods based on polymerase chain reaction have been developed for diagnostic purposes. Due to the expensive nature of the antibody drugs, identification of broad-spectrum antivirals is essential along with focusing on small interfering RNAs (siRNAs). High pathogenicity of NiV in humans, and lack of vaccines or therapeutics to counter this disease have attracted attention of researchers worldwide for developing effective NiV vaccine and treatment regimens.


Assuntos
Infecções por Henipavirus/veterinária , Vírus Nipah/imunologia , Vacinas Virais , Zoonoses , Animais , Doenças do Gato/epidemiologia , Doenças do Gato/prevenção & controle , Doenças do Gato/virologia , Gatos , Doenças do Cão/epidemiologia , Doenças do Cão/prevenção & controle , Doenças do Cão/virologia , Cães , Infecções por Henipavirus/epidemiologia , Infecções por Henipavirus/prevenção & controle , Infecções por Henipavirus/virologia , Humanos , Vírus Nipah/classificação , Vacinas Virais/administração & dosagem , Vacinas Virais/análise , Vacinas Virais/uso terapêutico , Zoonoses/epidemiologia , Zoonoses/prevenção & controle , Zoonoses/virologia
11.
Vet Res ; 50(1): 2, 2019 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-30616694

RESUMO

Porcine rotaviruses cause severe economic losses in the Korean swine industry due to G- and P-genotype mismatches between the predominant field and vaccine strains. Here, we developed a live attenuated trivalent porcine group A rotavirus vaccine using 80 cell culture passages of the representative Korean predominant strains G8P[7] 174-1, G9P[23] PRG942, and G5P[7] K71. Vaccination with the trivalent vaccine or its individual components induced no diarrhea during the first 2 weeks post-vaccination, i.e., the vaccines were attenuated. Challenge of trivalent-vaccinated or component-vaccinated piglets with homologous virulent strain(s) did not induce diarrhea for 2 weeks post-challenge. Immunization with the trivalent vaccine or its individual components also alleviated the histopathological lesions in the small intestines caused by challenge with the corresponding original virulent strain(s). Fecal secretory IgAs specific for each of vaccine strains were detected starting at 14 days post-vaccination (dpv), and IgA levels gradually increased up to 28 dpv. Oral immunization with the trivalent vaccine or its individual components induced high levels of serum virus-neutralizing antibody by 7 dpv. No diarrhea was observed in any experimental piglets during five consecutive passages of each vaccine strain. Our data indicated that the live attenuated trivalent vaccine was safe and effective at protecting piglets from diarrhea induced by challenge exposure of homologous virulent strains. This trivalent vaccine will potentially contribute toward controlling porcine rotavirus disease in South Korea and other countries where rotavirus infections with similar G and P genotypes are problematic.


Assuntos
Infecções por Rotavirus/veterinária , Rotavirus/imunologia , Doenças dos Suínos/prevenção & controle , Vacinas Virais/análise , Animais , República da Coreia , Infecções por Rotavirus/prevenção & controle , Suínos , Vacinas Atenuadas/análise
12.
J Virol Methods ; 268: 9-16, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30611776

RESUMO

Residual host cell DNA (rcDNA) from continuous cell lines used for manufacturing of biological medicinal products has been considered as safety risk. Historically, several analytical methods have been used for rcDNA quantitation including hybridization assay, Threshold® assay and quantitative polymerase chain reaction (qPCR). Sanofi Pasteur has a wealth of experience in the development of methods quantifying rcDNA in vaccines. Here, we compared the performance of our in-house assays for quantifying rcDNA in viral vaccines produced in Vero cells. Vero alpha-satellite sequence qPCR was compared with the hybridization and Threshold® assays in terms of specificity, sensitivity and precision. The impact of viral inactivation with ß-propiolactone (BPL) on rcDNA, within the vaccine production process, was also assessed. We demonstrate that the quantity of rcDNA measured is influenced by the analytical method used. Vero cell DNA-specific qPCR assay was shown to be robust with a large dynamic range and no matrix interference on a range of products. The qPCR assay demonstrated greater sensitivity and specificity versus the hybridization and Threshold® methods. Vero alpha-satellite sequence qPCR is a specific and sensitive method for the assessment of the quantity of Vero rcDNA in the highly purified vaccines.


Assuntos
Contaminação por DNA , DNA/análise , Vacinas Virais/análise , Virologia/métodos , Animais , Chlorocebus aethiops , Interações entre Hospedeiro e Microrganismos , Humanos , Células Vero
13.
Sci Rep ; 9(1): 720, 2019 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-30679646

RESUMO

Epstein-Barr virus (EBV), also known as human herpesvirus 4 (HHV-4), is a member of the Herpesviridae family and causes infectious mononucleosis, Burkitt's lymphoma, and nasopharyngeal carcinoma. Even in the United States of America, the situation is alarming, as EBV affects 95% of the young population between 35 and 40 years of age. In this study, both linear and conformational B-cell epitopes as well as cytotoxic T-lymphocyte (CTL) epitopes were predicted by using the ElliPro and NetCTL.1.2 webservers for EBV proteins (GH, GL, GB, GN, GM, GP42 and GP350). Molecular modelling tools were used to predict the 3D coordinates of peptides, and these peptides were then docked against the MHC molecules to obtain peptide-MHC complexes. Studies of their post-docking interactions helped to select potential candidates for the development of peptide vaccines. Our results predicted a total of 58 T-cell epitopes of EBV;  where the most potential were selected based on their TAP, MHC binding and C-terminal Cleavage score. The top most peptides were subjected to MD simulation and stability analysis. Validation of our predicted epitopes using a 0.45 µM concentration was carried out by using a systems biology approach. Our results suggest a panel of epitopes that could be used to immunize populations to protect against multiple diseases caused by EBV.


Assuntos
Epitopos de Linfócito T/imunologia , Infecções por Vírus Epstein-Barr/imunologia , Antígenos HLA/imunologia , Herpesvirus Humano 4/imunologia , Linfócitos T Citotóxicos/imunologia , Vacinas de Subunidades/imunologia , Proteínas Virais/imunologia , Biologia Computacional/métodos , Infecções por Vírus Epstein-Barr/prevenção & controle , Infecções por Vírus Epstein-Barr/virologia , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Glicoproteínas de Membrana/imunologia , Biologia de Sistemas/métodos , Vacinas de Subunidades/administração & dosagem , Vacinas de Subunidades/análise , Vacinas Virais/administração & dosagem , Vacinas Virais/análise , Vacinas Virais/imunologia
14.
Poult Sci ; 98(5): 1985-1992, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30566627

RESUMO

Newcastle disease virus (NDV)-attenuated vaccine has been widely used to prevent ND in poultry flocks, while many reports also mentioned the exogenous virus contamination in attenuated vaccines, which might be the reason for the widespread of some contagious diseases. Recently, the chicken infectious anemia virus (CIAV) contamination in the NDV-attenuated vaccine was also found in China, though no systemic study has studied the pathogenicity or infection mechanism of this special transmission route. Accordingly, simulation experiments were launched using CIAV isolated from a contaminated NDV-attenuated vaccine. Results showed that using NDV-attenuated vaccine contaminated with CIAV could cause CIA in chickens with obvious symptoms, including anemia, hemorrhage, lymphoatrophy, and growth retardation, while the synergistic reaction of CIAV and LaSota prompted their multiplication in vivo and disturbed the production of antibodies against each other. And CIAV could significantly reduce the NDV antibody titers and decrease the protective effectiveness. This study showed the synergetic pathogenicity of CIAV and LaSota strain after using contaminated NDV-attenuated vaccine, helping us to understand how the CIAV causes infection and induces severe diseases with a relatively low dose through the mouth, as well as reminding us that the damage of an attenuated vaccine contaminated with CIAV even in extremely low dose is not insignificant.


Assuntos
Vírus da Anemia da Galinha/patogenicidade , Galinhas , Vírus da Doença de Newcastle/patogenicidade , Vacinas Virais/análise , Animais , Anticorpos Antivirais/análise , Vírus da Anemia da Galinha/imunologia , China , Infecções por Circoviridae/veterinária , Infecções por Circoviridae/virologia , Doença de Newcastle/virologia , Vírus da Doença de Newcastle/imunologia , Doenças das Aves Domésticas/virologia , Vacinas Atenuadas/análise , Virulência
15.
Transbound Emerg Dis ; 65(4): 1103-1106, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29479824

RESUMO

One avian leukosis virus subgroup J (ALV-J) strain was isolated from 67 commercial live poultry vaccines produced by various manufacturers during 2013-2016 in China. The complete genomes of the isolate were sequenced and it was found that the genes gag and pol of the strain were relatively conservative, while the gp85 gene of the strain GX14YYA1 had the highest similarities with a field strain GX14ZS14, which was isolated from the chickens of a farm that had once used the same vaccine as the one found to be contaminated with the GX14YYA1. This is the first report of ALV-J contaminant in live poultry vaccine in China. Our finding demonstrates that vaccination of the commercial live vaccines might be a potential new route for ALV-J transmission in chickens and highlights the need for more extensive monitoring of the commercial live vaccines in China.


Assuntos
Vírus da Leucose Aviária/genética , Vírus da Leucose Aviária/isolamento & purificação , Leucose Aviária/transmissão , Contaminação de Medicamentos , Genoma Viral/genética , Doenças das Aves Domésticas/transmissão , Análise de Sequência de DNA/veterinária , Vacinas Virais/análise , Animais , Leucose Aviária/virologia , Sequência de Bases , Galinhas/virologia , China , Ensaio de Imunoadsorção Enzimática/veterinária , Reação em Cadeia da Polimerase/veterinária , Aves Domésticas , Doenças das Aves Domésticas/virologia , Análise de Sequência , Vacinação
16.
Curr Pharm Biotechnol ; 18(8): 638-647, 2017 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-28914197

RESUMO

BACKGROUND: Vaccine formulations may contain visible and/or subvisible particles, which can vary in both size and morphology. Extrinsic particles, which are particles not part of the product such as foreign contaminants, are generally considered undesirable and should be eliminated or controlled in injectable products. However, biological products, in particular vaccines, may also contain particles that are inherent to the product. Here we focus on the characterization of visible and subvisible particles in a live, replication-deficient viral vaccine candidate against HSV genital herpes in an early developmental stage. METHOD: HSV-2 viral vaccine was characterized using a panel of analytical methods, including Fourier transform infrared spectroscopy (FTIR), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blot, liquid chromatography-mass spectrometry (LC-MS), light microscopy, transmission electron microscopy (TEM), micro-flow imaging (MFI), dynamic light scattering (DLS), right angle light scattering (RALS), and intrinsic fluorescence. RESULTS: Particles in HSV-2 vaccine typically ranged from hundreds of nanometers to hundreds of micrometers in size and were determined to be inherent to the product. The infectious titer did not correlate with any trend in subvisible particle concentration and size distribution as shown by DLS, MFI, and TEM under stressed conditions. This suggested that particle changes in the submicron range were related to HSV-2 virion structure and had direct impact on biological activity. It was also observed that subvisible and visible particles could induce aggregation in the viral product. The temperature induced aggregation was observed by RALS, intrinsic fluorescence, and DLS. The increase of subvisible particle size with temperature could be fitted to a two-step thermokinetic model. CONCLUSION: Visible and subvisible particles were found to be inherent to the HSV-2 viral vaccine product. The mechanism of protein aggregation was discussed and a two-step thermokinetic aggregation profile was proposed. The approaches reported in this study may be applied to a variety of vaccines and other biological products, as a way to assess the consistency of the manufacturing process and identify key product quality attributes.


Assuntos
Composição de Medicamentos/métodos , Herpesvirus Humano 2/imunologia , Vacinas Virais/análise , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Eletroforese em Gel de Poliacrilamida , Congelamento , Herpesvirus Humano 2/química , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Agregados Proteicos , Estabilidade Proteica , Espectroscopia de Infravermelho com Transformada de Fourier , Vacinas Virais/normas , Vírion/ultraestrutura
17.
Poult Sci ; 96(5): 1094-1099, 2017 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-27794542

RESUMO

To investigate the possible causes of the massive spread of fowl adenovirus (FAdV) infection among chickens in recent years in China, 32 batches of live-virus vaccines were tested for contamination with FAdV by PCR. Among these, 1 live Newcastle disease virus (NDV) vaccine of the LaSota strain was demonstrated to be positive for contamination. The amplified hexon gene exhibited 99.8% identity with a recent Chinese field isolate (JSJ13) of FAdV-4. The positive LaSota vaccine was first neutralized with anti-NDV serum and then inoculated into specific pathogen-free embryos at embryonic day 5 through the yolk sac for isolation of the contaminated FAdV. The same hexon gene bands were amplified from extracted DNA of the liver tissues and chicken embryo allantoic fluid of the inoculated embryos, indicating the replication and isolation of the FAdV-4 virus strain that had contaminated the vaccine. This represents the first report of FAdV-4 contamination in a live vaccine for poultry in China. These findings suggest that contamination of live vaccine might represent one of the most important causes of massive outbreaks of FAdV infection among chickens and indicate that FAdV should therefore be included in the regular monitoring list for the detection of exogenous viral contamination of attenuated vaccines for poultry.


Assuntos
Infecções por Adenoviridae/veterinária , Adenoviridae/isolamento & purificação , Contaminação de Medicamentos , Doenças das Aves Domésticas/virologia , Vacinas Atenuadas/análise , Vacinas Virais/análise , Adenoviridae/genética , Infecções por Adenoviridae/etiologia , Animais , Embrião de Galinha , China/epidemiologia , Doença de Newcastle/prevenção & controle , Doenças das Aves Domésticas/etiologia , Análise de Sequência de DNA
18.
Pesqui. vet. bras ; 36(12): 1155-1159, Dec. 2016. tab
Artigo em Inglês | LILACS, VETINDEX | ID: biblio-842035

RESUMO

In order to investigate the immune enhancement effects of Ophiopogon japonicus polysaccharide Ophiopogon japonicus (OJPS) on Newcastle disease (ND) live vaccine, chickens vaccinated against ND live vaccine was orally administered with the OJPS at high, medium and low concentrations respectively. In negative control group, chickens were given orally equal volume of physiological saline. On day 14, 21 and 28, the serum antibody titer, erythrocyte-C3b receptor rosette rate (E-C3bRR), erythrocyte-C3b immune complex rosette rate (E-ICRR) and peripheral lymphocyte proliferation were measured. The results showed that at most time points, the antibody titer, peripheral lymphocyte proliferation, E-C3bRR and elimination rate of immune complex of three OJPS administrating groups were significantly higher (P<0.05) than those in negative control group. It indicated that OJPS could significantly improve the immune efficacy of Newcastle disease live vaccine, Ophiopogon japonicus polysaccharide possessed synergistical immunoenhancement.(AU)


Assuntos
Animais , Galinhas/virologia , Doença de Newcastle/imunologia , Ophiopogon/química , Vacinas Virais/análise , Adjuvantes Imunológicos , Anticorpos/sangue , Eritrócitos/imunologia , Linfócitos/imunologia
19.
J Gen Virol ; 97(11): 2809-2815, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27609617

RESUMO

Specific-pathogen-free (SPF) chickens were inoculated with the virus seed of an infectious bursal disease virus (IBDV)-attenuated vaccine, and positive reticuloendotheliosis virus (REV) antibody levels were subsequently detected in the chicken sera, indicating potential REV contamination of the vaccine. After neutralization with IBDV-positive blood serum, the vaccine was inoculated into DF-1 cells for REV isolation and identification. An REV strain, designated IBD-C1605, was identified using an immunofluorescence assay test. Three pairs of primers were employed for the amplification, cloning and sequencing of three overlapping fragments of the IBD-C1605 genome, and the whole-genome sequence of this isolate was obtained after gene assembly. The genome was 8362 base pairs (nt) in length and its homology with the nucleotide sequences of different reference strains varied between 94.2 and 99.2 %. Isolate IBD-C1605 was inoculated into 1-day-old SPF chickens to observe its pathogenicity. Infection with this organism slowed down the weight gain of SPF chickens and caused atrophy of their immune organs, such as the bursa of Fabricius and thymus gland. Furthermore, the chicken antibody levels decreased significantly after Newcastle disease virus and avian influenza virus subtype H9 vaccine immunization. This is the first report on the isolation and identification of REV from attenuated vaccine virus seeds in China, and is also the first study on the pathogenicity of REV from a contaminated vaccine in China. Our findings contribute towards a better understanding of the detrimental effects of vaccine contamination with exogenous viruses such as REV.


Assuntos
Infecções por Birnaviridae/veterinária , Contaminação de Medicamentos , Genoma Viral , Vírus da Doença Infecciosa da Bursa/imunologia , Doenças das Aves Domésticas/virologia , Vírus da Reticuloendoteliose/genética , Vírus da Reticuloendoteliose/patogenicidade , Infecções por Retroviridae/veterinária , Vacinas Virais/análise , Animais , Anticorpos Antivirais/imunologia , Infecções por Birnaviridae/imunologia , Infecções por Birnaviridae/prevenção & controle , Infecções por Birnaviridae/virologia , Galinhas , China , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/prevenção & controle , Vírus da Reticuloendoteliose/isolamento & purificação , Infecções por Retroviridae/imunologia , Infecções por Retroviridae/virologia , Organismos Livres de Patógenos Específicos , Vacinação , Vacinas Virais/genética , Vacinas Virais/imunologia
20.
Vaccine ; 34(28): 3317-23, 2016 06 14.
Artigo em Inglês | MEDLINE | ID: mdl-27171751

RESUMO

In this report we describe the detection and identification of Bluetongue virus (BTV) contaminations in commercial vaccines. BTV RNA was detected in vaccine batches of Lumpy skin disease (LSD) and Sheep pox (SP) using quantitative PCR (qPCR) for VP1 and NS3 genes. Both batches were positive for VP1 and NS3 in qPCR. The LSD vaccine-derived sample was positive for VP1 and VP2 in conventional PCR. The SP vaccine-derived sample was examined by amplification of VP1, VP4, VP6, VP7, NS2 and NS3 gene segments in conventional PCR. The SP vaccine-derived sample was further propagated in embryonated chicken eggs (ECE) and Vero cells. Preliminary sequence analysis showed that the LSD vaccine-derived sequence was 98-99% similar to BTV9. Analysis of the six genomic segments from the SP vaccine-derived isolate showed the highest similarity to BTV26 (66.3-97.8%). These findings are particularly important due to the effect of BTV on cattle and sheep, for which the vaccines are intended. They also demonstrate the necessity of rigorous vaccine inspection and strict vaccine production control.


Assuntos
Vírus Bluetongue/isolamento & purificação , Contaminação de Medicamentos , Vacinas Virais/análise , Animais , Embrião de Galinha , Chlorocebus aethiops , Genes Virais , Reação em Cadeia da Polimerase em Tempo Real , Células Vero
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