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Open Access Highly Accessed Research article

Sambucus nigra extracts inhibit infectious bronchitis virus at an early point during replication

Christie Chen1, David M Zuckerman2, Susanna Brantley1, Michka Sharpe1, Kevin Childress1, Egbert Hoiczyk2 and Amanda R Pendleton13*

Author Affiliations

1 Division of Natural Science and Mathematics, Oxford College of Emory University, Oxford, GA 30054, USA

2 W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins University School of Public Health, Baltimore, MD 21205, USA

3 Department of Mathematics, Sciences & Engineering, Amarillo College, Amarillo, TX 79178, USA

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BMC Veterinary Research 2014, 10:24  doi:10.1186/1746-6148-10-24

Published: 16 January 2014

Abstract

Background

Infectious bronchitis virus (IBV) is a pathogenic chicken coronavirus. Currently, vaccination against IBV is only partially protective; therefore, better preventions and treatments are needed. Plants produce antimicrobial secondary compounds, which may be a source for novel anti-viral drugs. Non-cytotoxic, crude ethanol extracts of Rhodiola rosea roots, Nigella sativa seeds, and Sambucus nigra fruit were tested for anti-IBV activity, since these safe, widely used plant tissues contain polyphenol derivatives that inhibit other viruses.

Results

Dose–response cytotoxicity curves on Vero cells using trypan blue staining determined the highest non-cytotoxic concentrations of each plant extract. To screen for IBV inhibition, cells and virus were pretreated with extracts, followed by infection in the presence of extract. Viral cytopathic effect was assessed visually following an additional 24 h incubation with extract. Cells and supernatants were harvested separately and virus titers were quantified by plaque assay. Variations of this screening protocol determined the effects of a number of shortened S. nigra extract treatments. Finally, S. nigra extract-treated virions were visualized by transmission electron microscopy with negative staining.

Virus titers from infected cells treated with R. rosea and N. sativa extracts were not substantially different from infected cells treated with solvent alone. However, treatment with S. nigra extracts reduced virus titers by four orders of magnitude at a multiplicity of infection (MOI) of 1 in a dose-responsive manner. Infection at a low MOI reduced viral titers by six orders of magnitude and pretreatment of virus was necessary, but not sufficient, for full virus inhibition. Electron microscopy of virions treated with S. nigra extract showed compromised envelopes and the presence of membrane vesicles, which suggested a mechanism of action.

Conclusions

These results demonstrate that S. nigra extract can inhibit IBV at an early point in infection, probably by rendering the virus non-infectious. They also suggest that future studies using S. nigra extract to treat or prevent IBV or other coronaviruses are warranted.

Keywords:
Infectious bronchitis virus; Coronavirus; Sambucus nigra; Nigella sativa; Rhodiola rosea