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Applied and Environmental Microbiology, August 2008, p. 4799-4805, Vol. 74, No. 15
0099-2240/08/$08.00+0     doi:10.1128/AEM.00246-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Microencapsulation of Bacteriophage Felix O1 into Chitosan-Alginate Microspheres for Oral Delivery

Yongsheng Ma,^1,2 Jennifer C. Pacan,² Qi Wang,^2* Yongping Xu,¹ Xiaoqing Huang,² Anton Korenevsky,³ and Parviz M. Sabour^2*

Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China,¹ Guelph Food Research Centre, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, Ontario, Canada,² Department of Microbiology, College of Biological Science, University of Guelph, Guelph, Ontario, Canada³

Received 28 January 2008/ Accepted 20 May 2008

This paper reports the development of microencapsulated bacteriophage Felix O1 for oral delivery using a chitosan-alginate-CaCl₂ system. In vitro studies were used to determine the effects of simulated gastric fluid (SGF) and bile salts on the viability of free and encapsulated phage. Free phage Felix O1 was found to be extremely sensitive to acidic environments and was not detectable after a 5-min exposure to pHs below 3.7. In contrast, the number of microencapsulated phage decreased by 0.67 log units only, even at pH 2.4, for the same period of incubation. The viable count of microencapsulated phage decreased only 2.58 log units during a 1-h exposure to SGF with pepsin at pH 2.4. After 3 h of incubation in 1 and 2% bile solutions, the free phage count decreased by 1.29 and 1.67 log units, respectively, while the viability of encapsulated phage was fully maintained. Encapsulated phage was completely released from the microspheres upon exposure to simulated intestinal fluid (pH 6.8) within 6 h. The encapsulated phage in wet microspheres retained full viability when stored at 4°C for the duration of the testing period (6 weeks). With the use of trehalose as a stabilizing agent, the microencapsulated phage in dried form had a 12.6% survival rate after storage for 6 weeks. The current encapsulation technique enables a large proportion of bacteriophage Felix O1 to remain bioactive in a simulated gastrointestinal tract environment, which indicates that these microspheres may facilitate delivery of therapeutic phage to the gut.

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* Corresponding author. Mailing address: Guelph Food Research Centre, Agriculture and Agri-Food Canada, 93 Stone Road West, Guelph, Ontario N1G 5C9, Canada. Phone: (519) 780-8021. Fax: (519) 829-2600. E-mail for Qi Wang: wangq{at}agr.gc.ca. E-mail for Parviz M. Sabour: sabourp{at}agr.gc.ca

Published ahead of print on 30 May 2008.

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Applied and Environmental Microbiology, August 2008, p. 4799-4805, Vol. 74, No. 15
0099-2240/08/$08.00+0     doi:10.1128/AEM.00246-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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