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Impact of Milk Components on Recovery of Viral RNA from MS2 Bacteriophage

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Abstract

Noroviruses are responsible for approximately 44 % of outbreaks involving dairy products for which causative agents are reported. Recovery of viruses from milk and dairy products is a difficult task. The role of different components of milk in the recovery of viral RNA was evaluated in this study. Four model milk formulations (A–D) were prepared by mixing different combinations of lactose, whey protein, casein, and fat in water. Each model formulation was spiked with five concentrations of bacteriophage MS2. The phenol-guanidine thiocyanate-chloroform protocol was used for extracting viral RNA from the model milk formulations and then extracted RNA was measured by a nanodrop spectrophotometer in ng/μl. The results showed that casein and whey protein had the highest negative impact on RNA yield, especially when the number of MS2 was less than 1.3 pfu/ml. The highest RNA recovery was obtained from the model milk formulation containing all four components; lactose, whey protein, casein, and fat. The amount of extracted RNA was closely correlated with the dry matter content of each formulation and the spiked concentration of coliphage using response surface modeling (R2:0.93). It was determined that milk fat is the most effective component in facilitating RNA extraction and the highest RNA yield can be achieved via elimination of whey protein and casein from milk by centrifugation at 40,000×g for 60 min. To achieve the highest viral RNA recovery efficiency by the proposed method, milk fat must be recombined with the supernatant of the centrifuged sample and then homogenized before performing the extraction protocol.

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References

  • Abbaszadegan, M., Huber, M. S., Gerba, C. P., & Pepper, I. L. (1993). Detection of enteroviruses in groundwater using PCR. Applied and Environment Microbiology, 59, 1318–1324.

    CAS  Google Scholar 

  • Abbaszadegan, M., Monteiro, P., Nwachuku, N., Alum, A., & Ryu, H. (2008). Removal of adenovirus, calicivirus, and bacteriophages by conventional drinking water treatment. Journal of Environmental Science and Health, 43, 171–177.

    Article  PubMed  CAS  Google Scholar 

  • Alum, A., Enriquez, C., & Gerba, C. P. (2011). Impact of irrigation method, soil and virus type on produce contamination. journal Food and Environmental Virology, 3, 78–85.

    Article  Google Scholar 

  • Atmar, R. L., Metcalf, T. G., Neill, N. H., & Estes, M. (1993). Detection of enteric viruses in oysters by using the polymerase chain reaction. Applied and Environment Microbiology, 59, 631–635.

    CAS  Google Scholar 

  • Beutler, E., Gelbart, J., & Kulh, W. (1990). Interference of heparine with polymerase chain reaction. Bio Techniques, 9, 166.

    CAS  Google Scholar 

  • Chomczynski, P., & Sacchi, N. (1987). Single-step method of RNA isolation by acid guanidinium thiocyanate–phenol–chloroform extraction. Analytical Biochemistry, 162, 156–159.

    Article  PubMed  CAS  Google Scholar 

  • Corroding, M., & Dalgleish, D. G. (1997). Isolate from industrial buttermilk: Emulsifying properties of materials derived from fat globule membrane. Journal of Agriculture and Food Chemistry, 45, 4595–4600.

    Article  Google Scholar 

  • Dewettinck, K., Rombaut, R., Thienport, N., Le, T., Messens, K., & Camp, J. V. (2007). Nutritional and technological aspects of milk fat globule membrane material (review). International Dairy Journal, 18, 436–457.

    Article  Google Scholar 

  • Dubois, E., Agier, R., Traore, O., Hennechart, C., Merle, G., Cruciere, C., et al. (2002). Modified concentration method for the detection of enteric virus on fruits and vegetables by reverse transcriptase-polymerase chain reaction or cell culture. Journal of Food Protection, 65, 1962–1969.

    PubMed  CAS  Google Scholar 

  • Eichler, D. (2006). Medical biochemistry (pp. 11–20). Sudbury: Jones and Bartlett.

    Google Scholar 

  • Environmental Protection Agency. (2001). Male-specific (F+) and somatic coliphage in water by two-step enrichment procedure. Method 1601. Washington, DC: Office of Water.

    Google Scholar 

  • Gouvea, V., Glass, R. I., Woods, P., Taniguchi, K., Klark, H. F., Forrester, B., et al. (1990). Polymerase chain reaction amplification and typing of rotaviruses nucleic acid from stool specimens. Journal of Clinical Microbiology, 28, 276–282.

    PubMed  CAS  Google Scholar 

  • Hadad, L. C. (2002). Nucleic acid isolation by adhering to hydrophobic solid phase and removing with nonionic surfactant. United States patent. 6383783.

  • Jiang, X., Wang, J., Graham, D. Y., & Eates, M. K. (1992). Detection of Norwalk virus in stool by polymerase chain reaction. Journal of Clinical Microbiology, 30, 2529–2534.

    PubMed  CAS  Google Scholar 

  • Kanno, C., Shimomura, Y., & Takano, E. (1991). Physicochemical properties of milk fat emulsions stabilized with bovine milk fat globule membrane. Journal of Food Science, 56, 1219–1223.

    Article  CAS  Google Scholar 

  • Langer, A. J., Ayers, T., Grass, J., Lynch, M., Angulo, F. J., & Mahon, B. E. (2012). Nonpasteurized dairy products, disease outbreaks, and state laws—United States, 1993–2006. Emerging Infectious Diseases, 18(3), 385–391.

    Article  PubMed  Google Scholar 

  • Lees, D. N., Henshilwood, K., & Dore, W. J. (1994). Development of a method for detection of enteric viruses in shellfish by PCR with polioviruses as a model. Applied and Environment Microbiology, 60, 2999–3005.

    CAS  Google Scholar 

  • Leggitt, P. R., & Jaykus, L. A. (2000). Detection methods for human enteric viruses in representative foods. Journal of Food Protection, 63, 1738–1744.

    PubMed  CAS  Google Scholar 

  • Michalski, M. C., Michel, F., Sainmont, D., & Briard, V. (2001). Apparent ζ potential as a tool to assess mechanical damages to the milk fat globule membrane. Colloids and Surface B: Biointerfaces, 23, 23–30.

    Article  Google Scholar 

  • Miller, G. J., Applegate, S. L., Radloff, H. D., & Nelms, G. E. (1980). Rapid extraction of milk fat. Journal of Dairy Science, 64, 1861–1862.

    Article  Google Scholar 

  • Morille, M., Passirani, C., Vonarbourg, A., Clavreul, A., & Benoit, J. P. (2008). Progress in developing cationic vectors for non-viral systemic gene therapy against cancer. Biomaterials, 29, 3477.

    Article  PubMed  CAS  Google Scholar 

  • Mullendore, J. L., Sobsey, M. D., & Shieh, Y. C. C. (2001). Improved method for the recovery of hepatitis A virus from oyster. Journal of Virological Methods, 94, 25–35.

    Article  PubMed  CAS  Google Scholar 

  • Richards, G. P. (2001). Enteric virus contamination of foods through industrial practices: a primer on intervention strategies. Journal of Industrial Microbiology and Biotechnology, 27, 117–125.

    Article  PubMed  CAS  Google Scholar 

  • Rock, C., Alum, A., & Abbaszadegan, M. (2010). PCR Inhibitor levels in concentrates of biosolids samples predicted by a new method based on excitation emission matrix (EEM) spectroscopy. Applied and Environment Microbiology, 76, 8102–8109.

    Article  CAS  Google Scholar 

  • Rombaut, R., & Dewettinck, K. (2006). Properties, analysis and purification of milk polar lipids. International Dairy Journal, 16, 1362–1373.

    Article  CAS  Google Scholar 

  • Shieh, Y. S. C., Calci, K. R., & Baric, R. S. (1999). A method to detect low levels of enteric viruses in contaminated oyster. Applied and Environment Microbiology, 65, 4709–4714.

    CAS  Google Scholar 

  • Shieh, Y. S. H., Wait, P., Tai, L., & Sobsey, M. D. (1995). Methods to remove inhibitors in sewage and other fecal wastes for enterovirus detection by the polymerase chain reaction. Journal of Virological Methods, 54, 51–66.

    Article  PubMed  CAS  Google Scholar 

  • Smallwood, I. (1996). Handbook of organic solvent properties. Butlerworth-Heinemann: Elsevier. 306.

    Google Scholar 

  • Swaisgood, H. E. (1996). Characteristics of milk. In P. F. Fox (Ed.), Food chemistry (3rd ed., pp. 841–878). New York: Marcel Dekkel.

    Google Scholar 

  • VanVoorthuizen, E. M., Ashbolt, N. J., & Schafer, A. I. (2001). Role of hydrophobic and electrostatic interactions for initial enteric virus retention by MF membranes. Journal of Membrane Science, 194, 69–79.

    Article  CAS  Google Scholar 

  • Wheeler, J., Bally, M., Zhang, Y., Reimer, D., Hope, M., Cullis, P., Schrrer, P. (2004). Lipid–nucleic acid particles prepared via a hydrophobic lipid–nucleic acid complex intermediate and use for gene transfer. European patent. EP0832271.

  • Wilde, J., Eiden, J., & Yolken, R. (1990). Removal of inhibitory substances from human fecal specimen for detection of group A rotaviruses by reverse transcriptase and polymerase chain reaction. Journal of Clinical Microbiology, 28, 1300–1307.

    PubMed  CAS  Google Scholar 

  • Wilson, I. G. (1997). Inhibition and facilitation of nucleic acid amplification. Applied and Environment Microbiology, 63, 3741–3751.

    CAS  Google Scholar 

  • Yavarmanesh, M., Abbaszadegan, M., Mortazavi, A., HabibiNajafi, M. B., Bassami, M., & Nassiri, M. R. (2010). Impact of milk components in recovery of the MS2 bacteriophage as an indicator of enteric viruses. Journal of Virological Methods, 168, 103–107.

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Food Science and Technology, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

    M. Yavarmanesh, A. Mortazavi & M. B. Habibi Najafi

  2. Civil, Environmental and Sustainable Engineering, National Science Foundation Water and Environmental Technology Center, Arizona State University, Glendale, AZ, USA

    M. Abbaszadegan & A. Alum

  3. Faculty of Veterinary Medicine and Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

    M. R. Bassami

  4. Department of Animal Science and Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran

    M. R. Nassiri

Authors
  1. M. Yavarmanesh
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  2. M. Abbaszadegan
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  3. A. Alum
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  4. A. Mortazavi
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  5. M. B. Habibi Najafi
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  6. M. R. Bassami
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  7. M. R. Nassiri
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Correspondence to M. Yavarmanesh.

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Yavarmanesh, M., Abbaszadegan, M., Alum, A. et al. Impact of Milk Components on Recovery of Viral RNA from MS2 Bacteriophage. Food Environ Virol 5, 103–109 (2013). https://doi.org/10.1007/s12560-013-9107-3

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  • DOI: https://doi.org/10.1007/s12560-013-9107-3

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