J Vet Sci.  2014 Sep;15(3):439-442. 10.4142/jvs.2014.15.3.439.

Quantitative measurement of influenza virus replication using consecutive bronchoalveolar lavage in the lower respiratory tract of a ferret model

• Lee DH
• Kim JI
• Lee JW
• Chung WH
• Park JK
• Lee YN
• Han JS
• Kim HY
• Lee SW
• Song CS

• KMID: 2155629
• DOI: http://doi.org/10.4142/jvs.2014.15.3.439

Abstract

The ferret is an established animal model of influenza virus infection. Although viral replication in the upper respiratory tract is usually measured with consecutively collected nasal washes, daily evaluation of viral replication in the lung is limited because a large numbers of ferrets need to be sacrificed at consecutive time points. To overcome this limitation, we performed a virus quantification assay using bronchoalveolar lavage (BAL) fluid. This non-invasive BAL technique allows consecutive quantification of virus replication in the lungs of living ferrets. Our method can be used for the longitudinal evaluation of virus tropism in the lower respiratory tract.

Keyword

bronchoalveolar lavage; ferret; infection; influenza

MeSH Terms

Animals
Bronchoalveolar Lavage/*veterinary
Disease Models, Animal
Female
Ferrets/*virology
Influenza A Virus, H3N2 Subtype/*physiology
Orthomyxoviridae Infections/*veterinary/virology
Respiratory System/*virology
Virus Replication/*physiology

Figure

• 

  Fig. 1 Collection of bronchoalveolar lavage fluid (BALF) from the ferret model. (A) A sterile 3.0-mm endotracheal tube was placed in the trachea using the lighted guide of a laryngoscope. (B) A 6 French feeding tube was inserted through the lumen of the endotracheal tube. After each administration of 10 mL sterile phosphate buffered saline, BALF was retrieved by gentle suction.

Reference

1. Belser JA, Katz JM, Tumpey TM. The ferret as a model organism to study influenza A virus infection. Dis Model Mech. 2011; 4:575–579.

2. Cox RJ, Major D, Hauge S, Madhun AS, Brokstad KA, Kuhne M, Smith J, Vogel FR, Zambon M, Haaheim LR, Wood J. A cell-based H7N1 split influenza virion vaccine confers protection in mouse and ferret challenge models. Influenza Other Respir Viruses. 2009; 3:107–117.

3. Ducatez MF, Webb A, Crumpton JC, Webby RJ. Long-term vaccine-induced heterologous protection against H5N1 influenza viruses in the ferret model. Influenza Other Respir Viruses. 2012; 7:506–512.

4. Enserink M. Pandemic influenza. Ferrets shed light on new virus's severity and spread. Science. 2009; 325:17.

5. Lee YN, Lee DH, Park JK, Yuk SS, Kwon JH, Nahm SS, Lee JB, Park SY, Choi IS, Song CS. Experimental infection and natural contact exposure of ferrets with canine influenza virus (H3N2). J Gen Virol. 2013; 94:293–297.

6. Maher JA, DeStefano J. The ferret: an animal model to study influenza virus. Lab Anim (NY). 2004; 33:50–53.

7. Maines TR, Jayaraman A, Belser JA, Wadford DA, Pappas C, Zeng H, Gustin KM, Pearce MB, Viswanathan K, Shriver ZH, Raman R, Cox NJ, Sasisekharan R, Katz JM, Tumpey TM. Transmission and pathogenesis of swine-origin 2009 A(H1N1) influenza viruses in ferrets and mice. Science. 2009; 325:484–487.

8. Munster VJ, de Wit E, van den Brand JMA, Herfst S, Schrauwen EJA, Bestebroer TM, van de Vijver D, Boucher CA, Koopmans M, Rimmelzwaan GF, Kuiken T, Osterhaus ADME, Fouchier RAM. Pathogenesis and transmission of swine-origin 2009 A(H1N1) influenza virus in ferrets. Science. 2009; 325:481–483.

9. Spackman E, Senne DA, Bulaga LL, Myers TJ, Perdue ML, Garber LP, Lohman K, Daum LT, Suarez DL. Development of real-time RT-PCR for the detection of avian influenza virus. Avian Dis. 2003; 47:3 Suppl. 1079–1082.

10. Svitek N, von Messling V. Early cytokine mRNA expression profiles predict Morbillivirus disease outcome in ferrets. Virology. 2007; 362:404–410.

11. Veldhuis Kroeze EJB, Stittelaar KJ, Teeuwsen VJ, Dijkshoorn ML, van Amerongen G, de Waal L, Kuiken T, Krestin GP, Hinkula J, Osterhaus ADME. Consecutive CT in vivo lung imaging as quantitative parameter of influenza vaccine efficacy in the ferret model. Vaccine. 2012; 30:7391–7394.

12. Veldhuis Kroeze EJB, van Amerongen G, Dijkshoorn ML, Simon JH, de Waal L, Hartmann IJC, Krestin GP, Kuiken T, Osterhaus ADME, Stittelaar KJ. Pulmonary pathology of pandemic influenza A/H1N1 virus (2009)-infected ferrets upon longitudinal evaluation by computed tomography. J Gen Virol. 2011; 92:1854–1858.