Influence of deoxynivalenol-contaminated feed on the immune response of pigs after PRRSV vaccination and infection
Alix PierronEleni VatziaMaria StadlerKerstin H. MairSelma SchmidtMelissa R. StasSophie DürlingerHeinrich KreutzmannChristian KnechtGyula BalkaJulia LaglerMarianne ZarubaTill RümenapfArmin SaalmüllerElisabeth MayerAndrea LadinigWilhelm Gerner
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The impact of the Fusarium mycotoxin deoxynivalenol (DON) on the immune response against porcine reproductive and respiratory syndrome virus (PRRSV) vaccination and infection was investigated. Forty-two weaned piglets were separated into seven groups and received three different diets: Low DON (1.09 ppm), High DON (2.81 ppm) or No DON. These three treatments were split further into either vaccinated (Ingelvac PRRSFLEX EU) and challenged with PRRSV 28 days post-vaccination, or only infected at day 28. A seventh group received no DON, no vaccination, and no infection. Two weeks after challenge infection, when pigs were euthanized, the number of IFN-γ producing lymphocytes in the blood of vaccinated animals was lower in pigs on High DON compared to animals on Low DON or No DON. Intracellular cytokine staining showed that vaccinated animals fed with the Low DON diet had higher frequencies of TNF-α/IFN-γ co-producing CD4+ T cells than the other two vaccinated groups, particularly in lung tissue. Vaccinated animals on High DON had similar viral loads in the lung as the non-vaccinated groups, but several animals of the Low DON or No DON group receiving vaccination had reduced titers. In these two groups, there was a negative correlation between lung virus titers and vaccine-specific TNF-α/IFN-γ co-producing CD4+ T cells located either in lung tissue or blood. These results indicate that after PRRSV vaccination and infection, high levels of DON negatively influence immune parameters and clearance of the virus, whereas low DON concentrations have immunomodulatory effects.The susceptibility of 2 strains (Oahu and Hyogo strains) of Aedes albopictus to oral infection with chikungunya virus was compared. These strains showed different infection threshold and final content of the virus. Oahu strain was much susceptible than the Hyogo strain. Virus titers in individual mosquito of the Hyogo strain on 14th day post-infection ranged continuously between (10)^1 and (10)^3 PFU per mosquito. On the contrary, those of the Oahu strain were fallen into 3 groups such as negative (non-infective), low virus titer ((10)^3-(10)^4 PFU) and high virus titer ((10)^6-(10)^7 PFU).
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Abstract In this assay, we developed and evaluated a multiplex PCR (mPCR) for its ability in detecting multiple infections of swine simultaneously. Four pairs of primers were used to detect five viruses. Specific primers were designed for classical swine fever virus (CSFV), African swine fever virus (ASFV) and pseudorabies (PRV). A pair of primers was designed prudently for two different types of porcine reproductive and respiratory syndrome virus that respectively were porcine reproductive and respiratory syndrome virus (PRRSV), highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). The detection limits of the mPCR were 1.09×10 4 , 1.50×10 3 , 2.10×10 3 , 1.30×10 3 and 8.97×10 2 copies/reaction for CSFV, ASFV, HP-PRRSV, PRRSV and PRV, respectively. A total of 49 clinical specimens were tested by the mPCR, and the result showed that co-infection by two or three viruses was 51%. In conclusion, the PCR is a useful tool for clinical diagnosis of not only single infections but also mixed infections in swines.
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A C6/36 cell culture persistently infected by dengue 2 virus was established and retained for over 20 weeks. No CPE was observed at any stage of infection. However, some giant cells up to 2- or 3-fold the diameter of normal cells appeared in the established culture. Extracellular virus titers fluctuated with a periodic tendency while the antigen level remained constant, suggesting some viral particles evolved into noninfectious defective interfering particles. Plaque size tended to change during the passage of cultures. The passage 8 virus mainly produced plaques relatively smaller than those produced by the passage 1 virus. Nevertheless, the plaque sizes produced by the passage 18 virus were somewhat mosaic. A temperature-sensitive mutant was detected in the virus released from passages 8 and 18. In addition, passages 8 and 18 produced the lowest virus titers. When persistently infected viruses were grown in Vero cells at 37°, at passage 1 there was a high titer which decreased 5 days postinoculation, whereas at passages 8 and 18 titers were undetectable until 4 days postinoculation. It seems that dengue 2 virus persistently infected in C6/36 cells could have been genetically altered. This will be tested by nucleic acid analysis of the viruses.
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To understand the distribution of influenza A H9N2 virus in chickens and men in Shenzhen area.Virus isolation was performed in embryonated hen s eggs with routine method. The antibody to H9N2 virus was detected with micro-hemagglutination inhibition (HI) test, then the results were checked by using the neutralization assay in MDCK cells.Totally 27 strains of influenza A H9N2 virus were isolated from chickens in farm markets in Shenzhen, whereas no H9N2 virus was isolated from men. Approximately 26% of human sera with the HI titers > or =20 to H9N2 virus were detected. However only 7% of chicken sera with the HI titers > or =20 to H9N2 virus were detected. Meanwhile the HI titer and (MGT) of antibody to H9N2 virus in human sera increased with age. It was also found that there was a close relationship between HI antibody titer to H9N2 virus in human sera and occupation.The distribution of influenza A H9N2 virus in chicken and men in Shenzhen was rather wide. The human H9N2 virus infection probably derived from chicken H9N2 virus.
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A set of two oligonucliotide primers were designed according to the reserved sequences of ORF6 and ORF7 of porcine reproductive and respiratory syndrome virus(PRRSV) strain VR-2322.A diagnostic RT-PCR assay has been developed against PRRSV RNA through optimizing the reaction condition.Pigs were challenged intranasally on 0 day old with 1 mL of PRRSV virulent strain containing 10~(5.6) TCID_(50)/mL.7 days after being challenged pigs were injected by specific serum of neutralizing titer of 1∶64.Serum was collected at weekly internal from 0 to 42 days post-infection and RT-PCR assay was applied for detection of PRRSV RNA in serum.The results showed that high titer neutralizing antibody could terminat the replication of PRRSV in infected porcines completely.
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