Therapeutic use of phage cocktail for controlling Escherichia coli O157:H7 in gastrointestinal tract of mice

• Published in: Journal of bioscience and bioengineering 2005-09, Vol.100 (3), p.280-287
• Main Authors: Tanji, Yasunori, Shimada, Takuya, Fukudomi, Hiroshi, Miyanaga, Kazuhiko, Nakai, Yutaka, Unno, Hajime
• Format: Article
• Language: English
• Subjects: Administration, Oral; AGENT PATHOGENE; Animals; BACTERIOFAGOS; Bacteriolysis; BACTERIOPHAGE; BACTERIOPHAGES; Biological and medical sciences; Biotechnology; Cattle; Cell Count; Coliphages - isolation & purification; DRUGS; ESCHERICHIA COLI; Escherichia coli Infections - therapy; Escherichia coli O157 - virology; Escherichia coli O157:H7; Feces - virology; Fundamental and applied biological sciences. Psychology; Gastrointestinal Tract - microbiology; MEDICAMENT; MEDICAMENTOS; Mice; ORGANISMOS PATOGENOS; PATHOGENS; Phage sp15; Probiotics - administration & dosage; Probiotics - therapeutic use
• ISSN: 1389-1723; 1347-4421
• DOI: 10.1263/jbb.100.280

To investigate the therapeutical use of phage mixture for controlling gastrointestinal Escherichia coli O157:H7 cells, in vitro and in vivo experiments were conducted. Three phages, SP15, SP21, and SP22 were selected from 26 phage stock screened from feces of stock animals and sewage influent. Addition of single or binary phage to the E. coli cell batch-culture reduced the turbidity of the culture. However, reascend of the turbidity due to the appearance of phage resistance cell was observed. On the other hand, addition of three phage mixture (SP15-21-22) did not produce reascend of culture turbidity under aerobic condition. Under anaerobic condition, slight reascend of culture turbidity was observed after SP15-21-22 addition. Chemostat continuous culture was operated under anaerobic condition to optimize the titer of phage cocktail and frequency of the addition for controlling E. coli cells. Five-log decrease of E. coli cell concentration after addition of phage cocktail of 10 9 Plaque forming unit (PFU)/ml was observed. However, reascend of cell concentration was observed after 1 d incubation. Repeated addition of phage cocktail was effective to reduce the cell concentration. Suspension of phage cocktail in the buffer containing 0.25% CaCO 3 neutralized 9 times much more buffer of pH 2. Based on this in vitro experiment, phage cocktail (SP15-21-22) suspended in the buffer containing 0.25% CaCO 3 was orally administrated to the mice in which E. coli O157:H7 cells was administrated in 2-d advance. E. coli and phage concentration in the feces was monitored for 9 d after phage addition. High titer of phage was detected in the feces when the phage cocktail administrated daily. E. coli O157:H7 concentration in the feces has been reduced according to the time period. However, difference of E. coli concentration in the feces of mice administrates with phage and in the control mice without phage addition became slight after 9-d test period. High titer of the phage settled down in the gastrointestinal tracts and reduced the concentration of E. coli cell. Repeated oral administration of SP15-21-22 was effective for rapid evacuation of E. coli O157:H7 from the feces and gastrointestinal tract of mice.