Aqueous Extracts of Hibiscus sabdariffa Calyces Decrease Hepatitis A Virus and Human Norovirus Surrogate Titers

Abstract

Hibiscus sabdariffa extract is known to have antioxidant, anti-diabetic, and antimicrobial properties. However, their effects against foodborne viruses are currently unknown. The objective of this study was to determine the antiviral effects of aqueous extracts of H. sabdariffa against human norovirus surrogates (feline calicivirus (FCV-F9) and murine norovirus (MNV-1)) and hepatitis A virus (HAV) at 37 °C over 24 h. Individual viruses (~5 log PFU/ml) were incubated with 40 or 100 mg/ml of aqueous hibiscus extract (HE; pH 3.6), protocatechuic acid (PCA; 3 or 6 mg/ml, pH 3.6), ferulic acid (FA; 0.5 or 1 mg/ml; pH 4.0), malic acid (10 mM; pH 3.0), or phosphate buffered saline (pH 7.2 as control) at 37 °C over 24 h. Each treatment was replicated thrice and plaque assayed in duplicate. FCV-F9 titers were reduced to undetectable levels after 15 min with both 40 and 100 mg/ml HE. MNV-1 was reduced by 1.77 ± 0.10 and 1.88 ± 0.12 log PFU/ml after 6 h with 40 and 100 mg/ml HE, respectively, and to undetectable levels after 24 h by both concentrations. HAV was reduced to undetectable levels by both HE concentrations after 24 h. PCA at 3 mg/ml reduced FCV-F9 titers to undetectable levels after 6 h, MNV-1 by 0.53 ± 0.01 log PFU/ml after 6 h, and caused no significant change in HAV titers. FA reduced FCV-F9 to undetectable levels after 3 h and MNV-1 and HAV after 24 h. Transmission electron microscopy showed no conclusive results. The findings suggest that H. sabdariffa extracts have potential to prevent foodborne viral transmission.

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References

  1. Ali, B. H., Al Wabel, N., & Blunden, G. (2005). Phytochemical, pharmacological and toxicological aspects of Hibiscus sabdariffa L.: A review. Phytotherapy Research, 19(5), 369–375. doi:10.1002/ptr.1628.

    CAS  Article  PubMed  Google Scholar 

  2. Blanton, L. H., Adams, S. M., Beard, R. S., Wei, G., Bulens, S. N., Widdowson, M. A., et al. (2006). Molecular and epidemiologic trends of caliciviruses associated with outbreaks of acute gastroenteritis in the United States, 2000–2004. Journal of Infectious Diseases, 193(3), 413–421. doi:10.1086/499315.

    Article  PubMed  Google Scholar 

  3. Borges, A., Ferreira, C., Saavedra, M. J., & Simoes, M. (2013). Antibacterial activity and mode of action of ferulic and gallic acids against pathogenic bacteria. [Research Support, Non-U.S. Gov’t]. Microbial Drug Resistance, 19(4), 256–265. doi:10.1089/mdr.2012.0244.

    CAS  Article  PubMed  Google Scholar 

  4. Chao, C. Y., & Yin, M. C. (2009). Antibacterial effects of roselle calyx extracts and protocatechuic acid in ground beef and apple juice. Foodborne Pathogens and Disease, 6(2), 201–206. doi:10.1089/fpd.2008.0187.

    CAS  Article  PubMed  Google Scholar 

  5. Chaudhary, R. K., & Andonov, A. P. (1992). Effect of ribavirin on hepatitis A virus replication in vitro. Canadian Journal of Infectious Diseases, 2, 67–70.

    Google Scholar 

  6. Chomnawang, M. T., Surassmo, S., Nukoolkarn, V. S., & Gritsanapan, W. (2005). Antimicrobial effects of Thai medicinal plants against acne-inducing bacteria. [Research Support, Non-U.S. Gov’t]. Journal of Ethnopharmacology, 101(1–3), 330–333. doi:10.1016/j.jep.2005.04.038.

    Article  PubMed  Google Scholar 

  7. Cowan, M. M. (1999). Plant products as antimicrobial agents. [Review]. Clinical Microbiology Reviews, 12(4), 564–582.

    PubMed Central  CAS  PubMed  Google Scholar 

  8. Hall, A. J., Wikswo, M. E., Pringle, K., Gould, L. H., Parashar, U. D., Division of Viral Diseases, National Center for Immunization and Respiratory Diseases, CDC, et al. (2014). Vital signs: foodborne norovirus outbreaks—United States, 2009–2012. MMWR Morbidity and Mortality Weekly Report, 63(22), 491–495.

    PubMed  Google Scholar 

  9. Higginbotham, K. L., Burris, K. P., Zivanovic, S., Davidson, P. M., & Stewart, C. N., Jr. (2014a). Aqueous extracts of Hibiscus sabdariffa calyces as an antimicrobial rinse on hot dogs against Listeria monocytogenes and methicillin-resistant Staphylococcus aureus. Food Control, 40, 274–277. doi:10.1016/j.foodcont.2013.12.011.

    Article  Google Scholar 

  10. Higginbotham, K. L., Burris, K. P., Zivanovic, S., Davidson, P. M., & Stewart, C. N., Jr. (2014b). Antimicrobial activity of Hibiscus sabdariffa aqueous extracts against Escherichia coli O157:H7 and Staphylococcus aureus in a microbiological medium and milk of various fat concentrations. [Research Support, Non-U.S. Gov’t]. Journal of Food Protection, 77(2), 262–268. doi:10.4315/0362-028X.JFP-13-313.

    Article  PubMed  Google Scholar 

  11. Kakkar, S., & Bais, S. (2014). A review on protocatechuic acid and its pharmacological potential. [Review]. ISRN Pharmacology, 2014, 952943. doi:10.1155/2014/952943.

    PubMed Central  Article  PubMed  Google Scholar 

  12. Katayama, S., Ohno, F., Yamauchi, Y., Kato, M., Makabe, H., & Nakamura, S. (2013). Enzymatic synthesis of novel phenol acid rutinosides using rutinase and their antiviral activity in vitro. Journal of Agriculture and Food Chemistry, 61(40), 9617–9622. doi:10.1021/jf4021703.

    CAS  Google Scholar 

  13. Lin, H. H., Huang, H. P., Huang, C. C., Chen, J. H., & Wang, C. J. (2005). Hibiscus polyphenol-rich extract induces apoptosis in human gastric carcinoma cells via p53 phosphorylation and p38 MAPK/FasL cascade pathway. [Research Support, Non-U.S. Gov’t]. Molecular Carcinogenesis, 43(2), 86–99. doi:10.1002/mc.20103.

    CAS  Article  PubMed  Google Scholar 

  14. Liu, K. S., Tsao, S. M., & Yin, M. C. (2005). In vitro antibacterial activity of roselle calyx and protocatechuic acid. Phytotherapy Research, 19(11), 942–945. doi:10.1002/ptr.1760.

    CAS  Article  PubMed  Google Scholar 

  15. McKay, D. L., Chen, C. Y., Saltzman, E., & Blumberg, J. B. (2010). Hibiscus sabdariffa L. tea (tisane) lowers blood pressure in prehypertensive and mildly hypertensive adults. [Randomized Controlled Trial Research Support, Non-U.S. Gov’t Research Support, U.S. Gov’t, Non-P.H.S.]. Journal of Nutrition, 140(2), 298–303. doi:10.3945/jn.109.115097.

    CAS  Article  PubMed  Google Scholar 

  16. Morton, J. (1987). Roselle. In Fruits of warm climates (pp. 281–286). Miami, FL: Creative Resource Systems, Inc.

  17. Oh, E.-G., Kim, K.-L., Shin, S., Son, K.-T., Lee, H.-J., Kim, T., et al. (2013). Antiviral activity of green tea catechins against feline calicivirus as a surrogate for norovirus. Food Science and Biotechnology, 22(2), 593–598. doi:10.1007/s10068-013-0119-4.

    CAS  Article  Google Scholar 

  18. Olaleye, M. (2007). Cytotoxicity and antibacterial activity of methanolic extract of Hibiscus sabdariffa. Journal of Medicinal Plants Research, 1, 9–13.

    Google Scholar 

  19. Povey, R. C. (1978a). In vitro antiviral efficacy of ribavirin against feline calicivirus, feline viral rhinotracheitis virus, and canine parainfluenza virus. American Journal of Veterinary Research, 39(1), 175–178.

    CAS  PubMed  Google Scholar 

  20. Povey, R. C. (1978b). Effect of orally administered ribavirin on experimental feline calicivirus infection in cats. American Journal of Veterinary Research, 39(8), 1337–1341.

    CAS  PubMed  Google Scholar 

  21. Ravishankar, S., Jaroni, D., Zhu, L., Olsen, C., McHugh, T., & Friedman, M. (2012). Inactivation of Listeria monocytogenes on ham and bologna using pectin-based apple, carrot, and hibiscus edible films containing carvacrol and cinnamaldehyde. [Research Support, Non-U.S. Gov’t Research Support, U.S. Gov’t, Non-P.H.S.]. Journal of Food Science, 77(7), M377–M382. doi:10.1111/j.1750-3841.2012.02751.x.

    CAS  Article  PubMed  Google Scholar 

  22. Saini, P., Gayen, P., Nayak, A., Kumar, D., Mukherjee, N., Pal, B. C., et al. (2012). Effect of ferulic acid from Hibiscus mutabilis on filarial parasite Setaria cervi: molecular and biochemical approaches. [Research Support, Non-U.S. Gov’t]. Parasitology International, 61(4), 520–531. doi:10.1016/j.parint.2012.04.002.

    CAS  Article  PubMed  Google Scholar 

  23. Scallan, E., Hoekstra, R. M., Angulo, F. J., Tauxe, R. V., Widdowson, M. A., Roy, S. L., et al. (2011). Foodborne illness acquired in the United States—Major pathogens. Emerging Infectious Diseases, 17(1), 7–15. doi:10.3201/eid1701.091101p1.

    PubMed Central  Article  PubMed  Google Scholar 

  24. Su, X., & D’Souza, D. H. (2011). Grape seed extract for control of human enteric viruses. Applied and Environment Microbiology, 77(12), 3982–3987. doi:10.1128/AEM.00193-11.

    CAS  Article  Google Scholar 

  25. Su, X., & D’Souza, D. H. (2013). Grape seed extract for foodborne virus reduction on produce. Food Microbiology, 34(1), 1–6. doi:10.1016/j.fm.2012.10.006.

    Article  PubMed  Google Scholar 

  26. Su, X., Howell, A. B., & D’Souza, D. H. (2010a). Antiviral effects of cranberry juice and cranberry proanthocyanidins on foodborne viral surrogates—A time dependence study in vitro. [Research Support, Non-U.S. Gov’t]. Food Microbiology, 27(8), 985–991. doi:10.1016/j.fm.2010.05.027.

    CAS  Article  PubMed  Google Scholar 

  27. Su, X., Howell, A. B., & D’Souza, D. H. (2010b). The effect of cranberry juice and cranberry proanthocyanidins on the infectivity of human enteric viral surrogates. Food Microbiology, 27(4), 535–540. doi:10.1016/j.fm.2010.01.001.

    CAS  Article  PubMed  Google Scholar 

  28. Sunday, O. A., Munir, A. B., Akeeb, O., Bolanle, A., & Badaru, S. O. (2010). Antiviral effect of Hibiscus sabdariffa and Celosia argentea on measles virus. African J Microbioloical Research, 4(4), 293–296.

    Google Scholar 

  29. Tsai, P.-J., McIntosh, J., Pearce, P., Camden, B., & Jordan, B. R. (2002). Anthocyanin and antioxidant capacity in Roselle (Hibiscus sabdariffa L.) extract. Food Research International, 35(4), 351–356. doi:10.1016/S0963-9969(01)00129-6.

    CAS  Article  Google Scholar 

  30. Xu, J., Wang, J., Deng, F., Hu, Z., & Wang, H. (2008). Green tea extract and its major component epigallocatechin gallate inhibits hepatitis B virus in vitro. Antiviral Research, 78(3), 242–249. doi:10.1016/j.antiviral.2007.11.011.

    CAS  Article  PubMed  Google Scholar 

  31. Yang, Y. S., Wang, C. J., Huang, C. N., Chen, M. L., Chen, M. J., & Peng, C. H. (2013). Polyphenols of Hibiscus sabdariffa improved diabetic nephropathy via attenuating renal epithelial mesenchymal transition. [Research Support, Non-U.S. Gov’t]. Journal of Agriculture and Food Chemistry, 61(31), 7545–7551. doi:10.1021/jf4020735.

    CAS  Article  Google Scholar 

  32. Yin, M. C., & Chao, C. Y. (2008). Anti-Campylobacter, anti-aerobic, and anti-oxidative effects of roselle calyx extract and protocatechuic acid in ground beef. International Journal of Food Microbiology, 127(1–2), 73–77. doi:10.1016/j.ijfoodmicro.2008.06.002.

    CAS  Article  PubMed  Google Scholar 

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Acknowledgments

Funding for this research was provided partly by the Professional Development Award to D. D’Souza from the UT-Graduate School and partly by the University of Tennessee-Institute of Agriculture (TEN #00391), which is gratefully acknowledged. The authors also acknowledge Dr. Dunlap at UT-Advanced Microscopy Center for his assistance with the TEM observation and Dr. Malcond Valladares for his assistance with the Hibiscus calyx extract preparations.

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Correspondence to Doris H. D’Souza.

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Joshi, S.S., Dice, L. & D’Souza, D.H. Aqueous Extracts of Hibiscus sabdariffa Calyces Decrease Hepatitis A Virus and Human Norovirus Surrogate Titers. Food Environ Virol 7, 366–373 (2015). https://doi.org/10.1007/s12560-015-9209-1

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Keywords

  • Hibiscus extracts
  • Murine norovirus (MNV-1)
  • Feline calicivirus (FCV-F9)
  • Hepatitis A virus (HAV)
  • Reduction