Bacteriophage Administration Reduces the Concentration of Listeria monocytogenes in the Gastrointestinal Tract and Its Translocation to Spleen and Liver in Experimentally Infected Mice
To investigate the efficacy of phage supplementation in reducing pathogen numbers, mice were treated via oral gavage with a Listeria monocytogenes phage preparation (designated ListShield) before being orally infected with L. monocytogenes. The concentrations of L. monocytogenes in the liver, spleen, and intestines were significantly lower ( ) in the phage-treated than in the control mice. Phage and antibiotic treatments were similarly effective in reducing the levels of L. monocytogenes in the internal organs of the infected mice. However, the significant weight loss detected in the control and antibiotic-treated groups was not observed in the infected, ListShield-treated mice. Long-term (90 days), biweekly treatment of uninfected mice with ListShield did not elicit detectable changes in the microbiota of their large intestines or deleterious changes in their health. Our data support the potential feasibility of using bacteriophages to control proliferation of L. monocytogenes in mice without affecting commensal microbiota composition. 1. Introduction Food borne bacterial pathogens remain a major health threat. Beyond reducing pathogen load at the source and in the final product, few measures are currently available to protect the human host. Side effects associated with long-term antibiotic treatment and the danger of emerging novel antibiotic resistance strains make an antibiotic-based prevention regimen unfeasible. In contrast, promising efforts are now being directed towards utilizing our own commensal microbiota to improve resistance to pathogens. The conventional approach aims at reshaping microbiota composition towards beneficial bacteria by adding live probiotic bacteria and/or by enhancing their growth through addition of prebiotic supplements [1]. An alternative approach for shaping overall microbiota composition aimed at reducing detrimental and potentially pathogenic bacteria by means of specific bacteriophages has to date received much less attention. The human gastrointestinal (GI) tract is colonized by an abundant and diverse microbiota that plays a significant role in mucosal protection, regulation of GI immune tolerance, digestion of complex macromolecules including mucus and fiber, and vitamin K synthesis [2, 3]. Numerous factors (e.g., age, antibiotic treatment, diet, psychological and physical stress, hormone levels, etc.) may lead to physiological disturbances in the gut’s microbiota [4]. Such alterations may contribute to many chronic and degenerative diseases, including Crohn’s disease, ulcerative colitis, rheumatoid arthritis,
References
[1]
J. A. Vanderhoof and R. J. Young, “Current and potential uses of probiotics,” Annals of Allergy, Asthma and Immunology, vol. 93, no. 5, pp. 33–37, 2004.
[2]
F. Guarner, “Enteric flora in health and disease,” Digestion, vol. 73, supplement 1, pp. 5–12, 2006.
[3]
F. Guarner and J.-R. Malagelada, “Gut flora in health and disease,” The Lancet, vol. 361, no. 9356, pp. 512–519, 2003.
[4]
G. C. O'Sullivan, P. Kelly, S. O'Halloran et al., “Probiotics: an emerging therapy,” Current Pharmaceutical Design, vol. 11, no. 1, pp. 3–10, 2005.
[5]
F. Salvini, L. Granieri, L. Gemmellaro, and M. Giovannini, “Probiotics, prebiotics and child health: where are we going?” Journal of International Medical Research, vol. 32, no. 2, pp. 97–108, 2004.
[6]
J. A. Hawrelak and S. P. Myers, “The causes of intestinal dysbiosis: a review,” Alternative Medicine Review, vol. 9, no. 2, pp. 180–197, 2004.
[7]
G. Perdigón, A. de Moreno de LeBlanc, J. Valdez, and M. Rachid, “Role of yoghurt in the prevention of colon cancer,” European Journal of Clinical Nutrition, vol. 56, supplement 3, pp. 65–68, 2002.
[8]
M. P. Taranto, G. Perdigón, M. Médici, and G. F. De Valdez, “Animal model for in vivo evaluation of cholesterol reduction by lactic acid bacteria,” Methods in Molecular Biology, vol. 268, pp. 417–422, 2004.
[9]
H. Brussow and E. Kutter, “Phage ecology,” in Bacteriophages: Biology and Application, E. Kutter and A. Sulakvelidze, Eds., pp. 129–163, CRC Press, Boca Raton, Fla, USA, 2005.
[10]
W. O. K. Grabow and P. Coubrough, “Practical direct plaque assay for coliphages in 100-ml samples of drinking water,” Applied and Environmental Microbiology, vol. 52, no. 3, pp. 430–433, 1986.
[11]
R. Armon and Y. Kott, “A simple, rapid and sensitive presence/absence detection test for bacteriophage in drinking water,” Journal of Applied Bacteriology, vol. 74, no. 4, pp. 490–496, 1993.
[12]
R. Armon, R. Araujo, Y. Kott, F. Lucena, and J. Jofre, “Bacteriophages of enteric bacteria in drinking water, comparison of their distribution in two countries,” Journal of Applied Microbiology, vol. 83, no. 5, pp. 627–633, 1997.
[13]
R. M. Araujo, A. Puig, J. Lasobras, F. Lucena, and J. Jofre, “Phages of enteric bacteria in fresh water with different levels of faecal pollution,” Journal of Applied Microbiology, vol. 82, no. 3, pp. 281–286, 1997.
[14]
O. Bergh, K. Y. Borsheim, G. Bratbak, and M. Heldal, “High abundance of viruses found in aquatic environments,” Nature, vol. 340, no. 6233, pp. 467–468, 1989.
[15]
F. Rohwer, “Global phage diversity,” Cell, vol. 113, no. 2, p. 141, 2003.
[16]
J. Lasobras, J. Dellunde, J. Jofre, and F. Lucena, “Occurrence and levels of phages proposed as surrogate indicators of enteric viruses in different types of sludges,” Journal of Applied Microbiology, vol. 86, no. 4, pp. 723–729, 1999.
[17]
H. Brüssow and R. W. Hendrix, “Phage genomics: small is beautiful,” Cell, vol. 108, no. 1, pp. 13–16, 2002.
[18]
M. Gautier, A. Rouault, P. Sommer, and R. Briandet, “Occurrence of Propionibacterium freudenreichii bacteriophages in Swiss cheese,” Applied and Environmental Microbiology, vol. 61, no. 7, pp. 2572–2576, 1995.
[19]
F.-C. Hsu, Y.-S. C. Shieh, and M. D. Sobsey, “Enteric bacteriophages as potential fecal indicators in ground beef and poultry meat,” Journal of Food Protection, vol. 65, no. 1, pp. 93–99, 2002.
[20]
J. E. Kennedy Jr., C. I. Wei, and J. L. Oblinger, “Methodology for enumeration of coliphages in foods,” Applied and Environmental Microbiology, vol. 51, no. 5, pp. 956–962, 1986.
[21]
J. E. Kennedy Jr., J. L. Oblinger, and G. Bitton, “Recovery of coliphages from chicken, pork sausage and delicatessen meats,” Journal of Food Protection, vol. 47, no. 8, pp. 623–626, 1984.
[22]
P. A. Whitman and R. T. Marshall, “Characterization of two psychrophilic pseudomonas bacteriophages isolated from ground beef,” Applied Microbiology, vol. 22, no. 3, pp. 463–468, 1971.
[23]
P. A. Whitman and R. T. Marshall, “Isolation of psychrophilic bacteriophage-host systems from refrigerated food products,” Applied Microbiology, vol. 22, no. 2, pp. 220–223, 1971.
[24]
M. Breitbart, I. Hewson, B. Felts et al., “Metagenomic analyses of an uncultured viral community from human feces,” Journal of Bacteriology, vol. 185, no. 20, pp. 6220–6223, 2003.
[25]
G. Bachrach, M. Leizerovici-Zigmond, A. Zlotkin, R. Naor, and D. Steinberg, “Bacteriophage isolation from human saliva,” Letters in Applied Microbiology, vol. 36, no. 1, pp. 50–53, 2003.
[26]
A. Sulakvelidze and E. Kutter, “Bacteriophage therapy in humans,” in Bacteriophages: Biology and Application, E. Kutter and A. Sulakvelidze, Eds., pp. 381–436, CRC Press, Boca Raton, Fla, USA, 2005.
[27]
A. Sulakvelidze, Z. Alavidze, and J. G. Morris Jr., “Bacteriophage therapy,” Antimicrobial Agents and Chemotherapy, vol. 45, no. 3, pp. 649–659, 2001.
[28]
A. Sulakvelidze and P. Barrow, “Phage therapy in animals and agribusiness,” in Bacteriophages: Biology and Applications, E. Kutter and A. Sulakvelidze, Eds., pp. 335–380, CRC Press, Boca Raton, Fla, USA, 2005.
[29]
C. R. Merril, D. Scholl, and S. L. Adhya, “The prospect for bacteriophage therapy in Western medicine,” Nature Reviews Drug Discovery, vol. 2, no. 6, pp. 489–497, 2003.
[30]
K. Ho, “Bacteriophage therapy for bacterial infections: rekindling a memory from the pre-antibiotics era,” Perspectives in Biology and Medicine, vol. 44, no. 1, pp. 1–16, 2001.
[31]
J. Alisky, K. Iczkowski, A. Rapoport, and N. Troitsky, “Bacteriophages show promise as antimicrobial agents,” Journal of Infection, vol. 36, no. 1, pp. 5–15, 1998.
[32]
W. C. Summers, “Bacteriophage therapy,” Annual Review of Microbiology, vol. 55, pp. 437–451, 2001.
[33]
A. Bruttin and H. Brüssow, “Human volunteers receiving Escherichia coli phage T4 orally: a safety test of phage therapy,” Antimicrobial Agents and Chemotherapy, vol. 49, no. 7, pp. 2874–2878, 2005.
[34]
E. Denou, A. Bruttin, C. Barretto, C. Ngom-Bru, H. Brüssow, and S. Zuber, “T4 phages against Escherichia coli diarrhea: potential and problems,” Virology, vol. 388, pp. 21–30, 2009.
[35]
M. H. Adams, Methods of Study Bacterial Viruses, Bacteriophages, Interscience Publishers, London, UK, 1959.
[36]
M. H. Adams, Enumeration of Bacteriophage Particles, Bacteriophages, Interscience Publishers, London, UK, 1959.
[37]
D. N. Miller, J. E. Bryant, E. L. Madsen, and W. C. Ghiorse, “Evaluation and optimization of DNA extraction and purification procedures for soil and sediment samples,” Applied and Environmental Microbiology, vol. 65, no. 11, pp. 4715–4724, 1999.
[38]
E. G. Zoetendal, A. D. L. Akkermans, and W. M. De Vos, “Temperature gradient gel electrophoresis analysis of 16S rRNA from human fecal samples reveals stable and host-specific communities of active bacteria,” Applied and Environmental Microbiology, vol. 64, no. 10, pp. 3854–3859, 1998.
