Document Type : Research Paper

Authors

1 Ph.D. Candidate, Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

2 Assistant Professor, Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

3 Professor, Department of Poultry Science, Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran

4 Associate Professor, Department of Virology, Faculty of Veterinary, Tehran University, Tehran, Iran

Abstract

Three experiments were carried out for isolation of bacteriophage against growth reducing Enterococcus faecalis and evaluation of its efficacy to control Enterococci colonization in intestine of Japanese quails. In experiment one, 60 Enterococci spp were isolated from gastrointestinal contents of adult quails, which among them seven isolates were selected based on preliminary tests. In experiment two, effect of oral challenge by the seven selected isolates along with Enterococcus faecalis ATCC51299- as positive control on growth performance of growing quails was evaluated. In experiment three, lytic bacteriophage was screened against Enterococcus faecalis using quails excreta and poultry house swage as a sources of phage. Finally, efficiency of bacteriophage therapy (104 pfu/mL) in biocontrol of enterococcal colonization of intestine was evaluated in quails orally challenged by Enterococcus faecalis (107 cfu/mL). Among seven isolates from the first experiment, an isolate which reduced the growth of growing quails and had the minimum mortality in comparison to negative control group (P<0.05) was final selected and identified at the species level using biochemical tests based of carbohydrates fermentation profile as Enterococcus faecalis. Oral administration of isolated bacteriophage against Enterococcus faecalis was able to reduce the intestinal enterococci colonization to a level comparable to negative control group (P<0.05). In conclusion, isolated bacteriophage against Enterococcus faecalis effectively reduced enterococci colonization in intestinal content of quails and in this way, a growth reducing factor in poultry could be biocontrolled.

Keywords

1 . Adams MH (1959) Bacteriophages. New York: International Science Publishers. Pp. 121- 132.
2 . Andreatti Filho RL, Higgins JP, Higgins SE, Gaona G, Wolfenden AD, Tellez G and Hargis BM (2007) Ability of bacteriophages isolated from different sources to reduce Salmonella enterica serovar Enteritidis in vitro and in vivo. Poultry Science. 86(9): 1904-1909.
3 . Atterbury RJ, Van Bergen MA, Ortiz F, Lovell MA, Harris JA, De BA, Wagenaar JA and Allen VM (2007) Bacteriophage therapy to reduce Salmonella colonization of broiler chickens. Applied Environmental Microbiology. 73: 4543-4549.
4 . Bachrach G, Zigmond ML, Zlotkin A, Naor R and Steinberg D (2003) Bacteriophage isolation from human saliva. Letters in Applied Microbiology. 36(1): 50-53.
5 . Barrow PA and Soothill JS (1997) Bacteriophage therapy and prophylaxis: rediscovery and renewed assessment of potential. Trends in Microbiology. 5: 268-271.
6 . Beaudoin RN, DeCesaro DR, Durkee DL and Barbaro SE (2007) Isolation of a bacteriophage from sewage sludge and characterization of its bacterial host cell. Rivier Academic. 3(1): 1-8.
7 . Biswas B, Adhya S, Washart P, Paul B, Trostel AN and Powell B (2002) Bacteriophage therapy rescues mice bacteraemia from a clinical isolate of vancomycin-resistant Enterococcus faecium. Infection and Immunity. 70(1): 204-210.
8 . Greer GG (2005) Bacteriophage control of foodborne bacteria. Food Protocol. 68: 1102-1111.
9 . Hudson JA, Billington C, Carey-Smith G and Greening G (2005) Bacteriophages as biocontrol agents in food. Food Protocol. 68: 426-437.
10 . Huff WE, Huff GR, Rath NC, Balog JM and Donoghue AM (2002) Prevention of Escherichia coli infection in broiler chickens with a bacteriophage aerosol spray. Poultry Science. 81: 1486-1491.
11 . Huff WE, Huff GR, Rath NC, Balog JM and Donoghue AM (2003) Bacteriophage treatment of a severe Escherichia coli respiratory infection in broiler chickens. Avian Disease. 47: 1399-1405.
12 . Huff WE, Huff GR, Rath NC, Balog JM and Donoghue AM (2005) Alternatives to antibiotics: Utilization of bacteriophage to treat colibacillosis and prevent foodborne pathogens. Poultry Science. 84: 655-659.
13 . Joerger RD (2003) Alternatives to antibiotics: bacteriocins, antimicrobial peptides and bacteriophages. Poultry Science. 82: 640-647.
14 . Letkiewicz S, Miedzybrodzki R, Fortuna W, Weber-Dabrowska B and Górskib A (2009) Eradication of Enterococcus faecalis by phage therapy in chronic bacterial prostatitis - Case report. Folia Microbiologica. 54(5): 457-461.
15 . Linden PK and Mille CB (1999) Vancomycin-resistant enterococci: the clinical effect of a common nosocomial pathogen. Diagnostic Microbiology and Infectious Disease. 33: 113-120.
16 . Loc Carrillo C, Atterbury RJ, el-Shibiny A, Connerton PL, Dillon E, Scott A and Connerton IF (2005) Bacteriophage therapy to reduce Campylobacter jejuni colonization of broiler chickens. Applied Environmental Microbiology. 71: 6554-6563.
17 . Lorch A (1999) Bacteriophages: an alternative to antibiotics? Biotechnology and Development Monitor. 39: 14-17.
18 . Margaret JS, Marina LB, Alexandru W, Rachel M and Durda S (2010) Arthritis and osteomyelitis associated with Enterococcus cecorum infection in broiler and broiler breeder chickens in Ontario, Canada. Veterinary Diagnostic Investigation. 22: 643-645.
19 . Ramin MNF, Mary DB and Michael WH (2010) Novel bacteriophages in Enterococcus spp. Current Microbiology. 60: 400-406.
20 . Sambrook J and Russell DW (2001) Molecular cloning: A laboratory manual. New York: Cold Spring Harbor Laboratory Press.
21 . Samli HE, Dezcan S, Koc F, Ozduven M, Okur AA and Senkoylu N (2010) Effects of Enterococcus faecium supplementation and floor type on performance, morphology of erythrocytes and intestinal microbiota in broiler chickens. British Poultry Science. 51(4): 564-568.
22 . Thomas D and Martin W (2014) Environmental responses and phage susceptibility in foodborne pathogens: implications for improving applications in food safety. Current Opinion in Biotechnology. 26: 45-49.
23 . Uchiyama J, Rashel M, Maeda Y, Takemura I, Sugihara S and Akechi K (2008) Isolation and characterization of a novel Enterococcus faecalis bacteriophage phi EF24C as a therapeutic candidate. FEMS Microbiology Letters. 278(2): 200-206.
24 . Wagenaar JA, Van Bergen MA, Mueller MA, Wassenaar TM and Carlton RM (2010) Phage therapy reduces Campylobacter jejuni colonization in broilers. Veterinary Microbiology. 109(3): 275-283.