In vivo therapeutic evaluation of phage cocktail and probiotic in reducing Salmonella infection in Broilers

TARUN  KUMAR; V S  RAJORA; NIDDHI  ARORA; DINESH  MITTAL; AMIT  PRASAD

doi:10.56093/ijans.v93i12.115002

Authors

TARUN KUMAR Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125 001 India
V S RAJORA College of Veterinary and Animal Sciences, Govind Ballabh Pant University of Agriculture and Technology Pantnagar, Uttarakhand
NIDDHI ARORA College of Veterinary and Animal Sciences, Govind Ballabh Pant University of Agriculture and Technology Pantnagar, Uttarakhand
DINESH MITTAL Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana 125 001 India
AMIT PRASAD College of Veterinary and Animal Sciences, Govind Ballabh Pant University of Agriculture and Technology Pantnagar, Uttarakhand

https://doi.org/10.56093/ijans.v93i12.115002

Keywords:

Bacteriophages, Cocktail, Phage, Poultry, Probiotic, Salmonella, Therapeutic

Abstract

Antimicrobial resistance and ban on the usage of most of the antibiotics in food producing animals especially in poultry and pigs propelled research towards antibiotic alternatives. Food borne and multidrug-resistant (MDR) pathogens like Salmonella can enter food chain via consumption of the contaminated meat. Bacteriophage (phage) therapy could be used during rearing or pre-harvest stages of poultry production to overcome these emerging problems. The present study was conducted to determine the therapeutic effectiveness of bacteriophage cocktail and probiotics against Salmonella gallinarum in experimentally infected broiler chicks based on mortality, clinical manifestations, and faecal shedding. In vitro evaluation revealed that optical density (OD) of Salmonella gallinarum at MOIs of 102 was significantly reduced by individual phages as well as phage cocktail, with comparably less decrease in optical density in the culture treated with individual phage than the phage cocktail. Bacteriophage cocktail with concentration 1011 PFU/ml was able to protect 100% birds infected with Salmonella gallinarum. Faecal shedding rate was significantly low in the birds treated with bacteriophage cocktail and probiotics (1.2%) than untreated group. Significant improvement in body weight was seen in the bacteriophage treated groups as compared to untreated infected group. Based on the findings of the current study, administering high titre bacteriophages alone or in combination with probiotics for the effective management of Salmonella infection in broiler chicks may be suggested as an alternative to antibiotics as well as a useful strategy to control food borne pathogens in the food chain.

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References

Atterbury R J, Van Bergen M A P, Ortiz F, Lovell M A, Harris J A, De Boer A, Wagenaar J A, Allen V M and Barrow P A. 2007. Bacteriophage therapy to reduce Salmonella colonization of broiler chickens. Applied and Environmental Microbiology 73: 4543–549.

Ayed M H, Laamari Z and Rekik B. 2004. Effects of incorporating an antibiotic “avilamycin” and a probiotic “activis” in broiler diets. American Society of Animal Science 55: 237–240.

Connerton P L and Connerton I F. 2005. Microbial treatments to reduce pathogens in poultry meat, pp. 414-427. Food Safety Control in the Poultry Industry. (Ed) Mead G. Woodhead Publishing Ltd., Cambridge.

Costa P, Pereira C, Gomes Ana T P C and Almeida A. 2019. Efficiency of single phage suspensions and phage cocktail in the inactivation of Escherichia coli and Salmonella typhimurium: An in vitro preliminary study. Microorganisms 7(94): 1–17.

Dorea F C, Cole D J, Hofacre C, Zamperini K, Mathis D, Doyle M P, Lee M D and Maurer J J. 2010. Effect of vaccination of breeder chickens on contamination of broiler chicken carcasses in integrated poultry operations. Applied and Environmental Microbiology 76(23): 782–99.

FAO/WHO Expert Consultation. 2001. Evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. -http://www.who.int/ foodsafety/publications/fs management/en/probiotics.pdf.

Fischer S, Kittler S, Klein G and Glünder G. 2013. Impact of a single phage and a phage cocktail application in broilers on reduction of Campylobacter jejuni and development of resistance. PLoS ONE 8(10): e78543.

Greenwood P E and Nikulin M S. 1996. A Guide to Chi-Square Testing. Wiley, New York.

Huff W E, Huff G R, Rath N C, Balog J M and Donoghue A M. 2005. Alternatives to antibiotics: Utilization of bacteriophage to treat Colibacillosis and prevent foodborne pathogens. Poultry Science 84: 655–59.

Kabir S M L. 2009. The role of probiotics in the poultry industry. International Journal of Molecular Sciences 10: 3531–546.

Kumar T, Rajora V S, Arora N, Prakash A and Shukla S K. 2017. Isolation and characterization of Salmonella bacteriophages from poulty and pig sewage wastes. Indian Journal of Animal Sciences 87(5): 562–67.

O’Flynn G, Ross R P, Fitzgeral G F and Coffey A. 2004. Evaluation of a cocktail of three Bacteriophages for biocontrol of Escherichia coli O157:H7. Applied and Environmntal. Microbiology 70: 3417-3424.

Rohwer F and Edwards R. 2002. The phage proteomic tree: A genome-based taxonomy for phage. Journal of Bacteriology 184: 4529–535.

Saarela M, Mogensen G, Fondens R, Matto J and Mattila-Sandholm T. 2000. Probiotic bacteria: Safety, functional and technological properties. Journal of Biotechnology 84: 197–215.

Salminen S J, Nybom S, Meriluoto J, Collado M C, Vesterlund S and El-Nezami H. 2010. Interaction of probiotics and pathogens benefits to human health?. Current Opinion in Biotechnology 21: 157–67.

Shott S. 1990. Statistical for Health Professionals. W.B. Saunders Co. Philadelphia, pp. 313–336.

Soomro A H, Masud T and Rathore H A. 2002. Application of probiotic culture. Journal of Animal and Veterinary Advances 1: 40–2.

Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M, Monnet D L, Pulcini C, Kahlmeter G, Kluytmans J, Carmeli Y, Ouellette M, Outterson K, Patel J, Cavaleri M, Cox E M, Houchens C R, Grayson M L, Hansen P, Singh N, Theuretzbacher U and Magrini N. 2018. WHO pathogens priority list working group. Discovery, research, and development of new antibiotics: The WHO priority list of antibiotic-resistant bacteria and tuberculosis. The Lancet Infectious Diseases 18(3): 318–27.

Thanki A M, Clavijo V, Healy K, Wilkinson R C, Sicheritz-Pontén T, Millard A D and Clokie M R J. 2022. Development of a phage cocktail to target Salmonella strains associated with swine. Pharmaceuticals 15(1): 58.

Thanki A M, Hooton S, Whenham N, Salter M G, Bedford M R, O’Neill H V M and Clokie M R J. 2023. A bacteriophage coc ktail delivered in feed significantly reduced Salmonella colonization in challenged broiler chickens. Emerging Microbes and Infections 12(1): 2217947.

Timmerman H M, Veldman A, Van den Elsen E, Rombouts F M and Beynen A C. 2006. Mortality and growth performance of broilers given drinking water supplemented with chicken-specific probiotics. Poultry Science 85: 1383–388.

Turki Y, Ouzari H, Mehri I, Ammar A B and Hassen A. 2012. Evaluation of a cocktail of three bacteriophages for the biocontrol of Salmonella of wastewater. Food Research International 45: 1099–1105.

Wafaa A A, Madian K, Ebtehal A and Gehan M K. 2006. The effect of combined competitive exclusionvculture with mannan-oligosacharides and ciprofloxacin on Salmonella enteritidis colonization in broiler chickens. 12th Congress of Faculty of Vetertinary Medicine, Assiut University, Egypt.

Wagenaar J A, Van Bergen M A, Mueller M A, Wassenaar T M and Carlton R M. 2005. Phage therapy reduces Campylobacter jejuni colonization in broilers. Veterinary Microbiology 109: 275–83.