Chitosan Supplementation Reduces Enteric Colonization of Campylobacter jejuni in Broiler Chickens and Down-Regulates Expression of Colonization Genes

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Authors

  • Hanna R. Arambel
  • Ann M. Donoghue
  • Komala Arsi
  • Abhinav Upadhyay
  • Ann Woo-Ming
  • Pamela J. Blore
  • Kumar Venkitanarayanan
  • Dan J. Donoghue

DOI:

https://doi.org/10.17140/aftnsoj-1-119

Keywords:

Campylobacter jejuni, Chitosan, Broiler chickens, Pre-harvest, Colonization gene, Real-time quantitative PCR

Abstract

Campylobacter is one of the leading causes of foodborne bacterial gastroenteritis worldwide, and poultry is considered as the most common source of human infections. Cam-pylobacter is prevalent in most poultry flocks and a reduction of Campylobacter in poultry would greatly reduce the risk of campylobacteriosis in humans. Unfortunately, efforts to reduce Campylobacter in pre-harvest poultry have been met with limited success. Preliminary studies with the natural compound, chitosan, demonstrate its ability to kill Campylobacter, in vitro. The purpose of this study was to determine the ability of feed supplemented chitosan to reduce enteric Campylobacter colonization in broiler chickens. Additionally, the effect of chitosan on expression of Campylobacter’s chicken colonization genes was investigated using real-time quantitative PCR (RT-qPCR). Because chitosan’s antimicrobial properties may vary depending upon its molecular weight, selected doses of three molecular weight chitosans were supple-mented in the feed and evaluated for efficacy to reduce Campylobacter in chickens. Three replicate trials were conducted, and in each trial, birds were divided into 10 treatments (n=10 birds/treatment) and were fed 0% (controls), 0.25%, 0.5% or 1% (wt./wt.) of a low, medium or high molecular weight chitosan. Birds were fed treated feed for the duration of the study and orally challenged with a four-strain mixture of wild type C. jejuni on day 6. On day 15, the ceca samples were collected for enumeration of Campylobacter. In all three trials, the 0.5% dose of the medium molecular weight chitosan reduced cecal Campylobacter counts (P<0.05). RT-qPCR analysis revealed that chitosan down-regulated the expression of chicken colonization genes as compared to control (P<0.05). These results suggest that supplementation of chitosan in feed is a potential strategy to reduce the enteric colonization of Campylobacter in pre-harvest chickens.

References

Centers for Disease Control and Prevention. Campylobacter general information. Website: http://www.cdc.gov/nczved/divi-sions/dfbmd/diseases/campylobacter/ 2014; Accessed Septem-ber 15, 2015.

World Health Organization. The global view of cam-pylobacteriosis. Website: http://apps.who.int/iris/bitstre am/10665/80751/1/9789241564601_eng.pdf 2013; Accessed September 15, 2015.

Hermans D, Pasmans F, Messens W, et al. Poultry as a host for the zoonotic pathogen Campylobacter jejuni. Vector-Borne and Zoonotic Dis. 2012; 12(2): 89-98. doi: 10.1089/vbz.2011.0676

Sasaki Y, Haruna M, Mori T, et al. Quantitative estimation of Campylobacter cross-contamination in carcasses and chick-en products at an abattoir. Food Control. 2014; 43: 10-17. doi: 10.1016/j.foodcont.2014.02.015

Stern NJ, Fedorka-Cray P, Bailey JS, et al. Distribution of Campylobacter spp. in selected US poultry production and pro-cessing operations. J. Food Prot. 2001; 64(11): 1705-1710.

Hermans D, Van Deun K, Messens W, et al. Campylobacter control in poultry by current intervention measures ineffective: urgent need for intensified fundamental research. Vet Microbiol. 2011; 152(3): 219-228. doi: 10.1016/j.vetmic.2011.03.010

Menconi A, Hernandez-Velasco X, Latorre JD, et al. Effect of chitosan as a biological sanitizer for Salmonella Typhimurium and aerobic Gram negative spoilage bacteria present on chick-en skin. Int J Poult Sci. 2013; 12(6): 318-321. doi: 10.3923/ ijps.2013.318.321

No HK, Park NY, Lee SH, Meyers SP. Antibacterial activ-ity of chitosans and chitosan oligomers with different molecular weights. Int J of Food Microbiol. 2002; 74: 65-72. doi: 10.1016/ S0168-1605(01)00717-6

Rabea EI, Badawy ME, Stevens CV, Smagghe G, Steurbaut W. Chitosan as animicrobial agent: applications and mode of ac-tion. Biomacromolecules. 2003; 4(6): 1457-1465. doi: 10.1021/ bm034130m

No HK, Meyers SP. Preparation and characterization of chi-tin and chitosan-a review. J Aquat Food Prod Technol. 1995; 4(2): 27-52. doi: 10.1300/J030v04n02_03

No HK, Meyers SP, Prinyawiwatkul W, Xu Z. Applications of chitosan for improvement of quality and shelf life of foods: a review. J Food Sci. 2007; 72: R87-R100. doi: 10.1111/j.1750- 3841.2007.00383.x

Sagoo S, Board R, Roller S. Chitosan inhibits growth of spoilage micro-organisms in chilled pork products. Food Micro-biol. 2002; 19(2): 175-182. doi: 10.1006/fmic.2001.0474

Beverlya RL, Janes ME, Prinyawiwatkula W, No HK. Ed-ible chitosan films on ready-to-eat roast beef for the control of Listeria monocytogenes. Food Microbiol. 2008; 25(3): 534-537. doi: 10.1016/j.fm.2007.11.002

Helander IM, Nurmiaho-Lassila E-L, Ahvenainen R, Rhoades J, Roller S. Chitosan disrupts the barrier properties of the outer membrane of Gram-negative bacteria. Int J Food Micribiol. 2001; 71(2): 235-244.

Solis de los Santos F, Donoghue AM, Venkitanarayanan K, et al. Therapeutic supplementation of caprylic acid in feed re-duces Campylobacter jejuni colonization in broiler chicks. Appl Environ Microbiol. 2008; 74(14): 4564-4566. doi: 10.1128/ AEM.02528-07

Solis de los Santos F, Donoghue AM, Venkitanarayanan K, et al. The natural feed additive caprylic acid decreases Campylo-bacter jejuni colonization in market-aged broiler chickens. Poult Sci. 2009; 88(1): 61-64. doi: 10.3382/ps.2008-00228

Farnell MB, Donoghue AM, Cole K, Reyes-Herrera I, Blore PJ, Donoghue DJ. Campylobacter susceptibility to ciprofloxa-cin and corresponding fluoroquinolone concentrations within the gastrointestinal tracts of chickens. J Appl Microbiol. 2005; 99(5): 1043-1050. doi: 10.1111/j.1365-2672.2005.02712.x

Ganan M, Carrascosa V, Martínez-Rodríguez AJ. Antimicro-bial activity of chitosan against Campylobacter spp. and other microorganisms and its mechanism of action. J. Food Prot. 2009; 72(8): 1735-1738.

Line JE. Development of a selective differential agar for isolation and enumeration of Campylobacter spp. J Food Prot. 2001; 64(11): 1711-1715.

Arsi K, Donoghue AM, Venkitanarayanan K, et al. The ef-ficacy of the natural plant extracts, thymol and carvacrol against Campylobacter colonization in broiler chickens. J Food Safety. 2014; 34(4): 321-325. doi: 10.1111/jfs.12129

Upadhyay A, Johny AK, Amalaradjou MAR, Baskaran SA, Kim KS, Venkitanarayanan K. Plant-derived antimicrobi-als reduce Listeria monocytogenes virulence factors in vitro, and down-regulate expression of virulence genes. Int. J.Food Microbiol. 2012; 157(1): 88-94. doi: 10.1016/j.ijfoodmi-cro.2012.04.018

Byrd JA, Anderson RC, Callaway TR, et al. Effect of experi-mental chlorate product administration in the drinking water on Salmonella Typhimurium contamination of broilers. Poult Sci. 2003; 82(9): 1403-1406. doi: 10.1093/ps/82.9.1403

SAS Institute. SAS/STAT® Users guide: Release 9.2 edition. SAS Institute, Cary, NC, 2002.

Goh EB, Yim G, Tsui W, McClure J, Surette MG, Davies J. Transcriptional modulation of bacterial gene expression by sub-inhibitory concentrations of antibiotics. Proc Natl Acad Sci USA. 2002; 99(26): 17025-17030.

Fonseca AP, Extremina C, Fonseca AF, Sousa JC. Effect of sub-inhibitory concentration of piperacillin/tazobactam on Pseudomonas aeruginosa. J Med Microbiol. 2004; 53(9): 903- 910. doi: 10.1099/jmm.0.45637-0

Upadhyay A, Upadhyaya I, Kollanoor-Johny A, Venkitana-rayanan, K. Anti-biofilm effect of plant derived antimicrobials on Listeria monocytogenes. Food Microbiol. 2013; 36(1): 79-89. doi: 10.1016/j.fm.2013.04.010

Hermans D, Van Deun K, Martel A, et al. Colonization fac-tors of Campylobacter jejuni in the chicken gut. Vet Res. 2011; 42(1): 82. doi: 10.1186/1297-9716-42-82

Fernando U, Biswas D, Allan B, Willson P, Potter AA. Influ-ence of Campylobacter jejuni fliA, rpoN and flgK genes on colo-nization of the chicken gut. Int. J Food Microbiol. 2007; 118(2): 194-200. doi: 10.1016/j.ijfoodmicro.2007.07.038

Konkel ME, Gray SA, Kim BJ, Garvis SG, Yoon J. Identifi-cation of the enteropathogens Campylobacter jejuni and Campy-lobacter coli based on the cadF virulence gene and its product. J. Clin. Microbiol. 1999; 37(3): 510-517.

Metcalf JH, Donoghue AM, Venkitanarayanan K, et al. Water administration of the medium-chain fatty acid caprylic acid produced variable efficacy against enteric Campylobacter colonization in broilers. Poult. Sci. 2011; 90(2): 494-497. doi: 10.3382/ps.2010-00891

Hakkinen M, Schneitz C. Efficacy of a commercial competi-tive exclusion product against Campylobacter jejuni. Br Poult Sci. 1999; 40(5): 619-621. doi: 10.1080/00071669986990

Hilmarsson H, Thormar H, Thrainsson JH, et al. Effect of glycerol monocaprate (monocaprin) on broiler chickens: An at-tempt at reducing intestinal Campylobacter infection. Poult Sci. 2006; 85(4): 588-592. doi: 10.1093/ps/85.4.588

Robyn J, Rasschaert G, Hermans D, Pasmans F, Heyndrickx M. Is allicin able to reduce Campylobacter jejuni colonization in broilers when added to drinking water? Poult Sci. 2013; 92(5): 1408-1418. doi: 10.3382/ps.2012-02863

Carvalho CM, Gannon BW, Halfhide DE, et al. The in vivo efficacy of two administration routes of a phage cocktail to reduce numbers of Campylobacter coli and Campylobacter jejuni in chickens. BMC Microbiol. 2010; 10(1): 232. doi: 10.1186/1471-2180-10-232

Molatová Z, Skrivanova E, Baré J, Houf K, Bruggeman G, Marounek M. Effect of coated and non-coated fatty acid sup-plementation on broiler chickens experimentally infected with Campylobacter jejuni. J Anim Physiol Anim Nutr. 2010; 95(6): 701-706. doi: 10.1111/j.1439-0396.2010.01100.x

Van Deun K, Haesebrouck F, Van Immerseel F, Ducatelle R, Pasmans F. Short chain fatty acids and lactate as feed additives to control Campylobacter jejuni infections in broilers. Avian Path. 2008; 37(4): 379-383. doi: 10.1080/03079450802216603

Neal-McKinney JM, Lu X, Duong T, et al. Production of organic acids by probiotic Lactobacilli can be used to reduce pathogen load in poultry. PLoS One. 2012; 7(9): 1-11. doi: 10.1371/journal.pone.0043928

Chandumpai A, Singhpibulporn N, Faroongsarng D, Sorn-prasit P. Preparation and physico-chemical characterization of chitin and chitosan from the pens of the squid species, Lo-ligo lessoniana and Loligo formosana. Carbohydrate Polymers. 2004; 58(4): 467-474. doi: 10.1016/j.carbpol.2004.08.015

Qin C, Li H, Xiao Q, Liu Y, Zhu J, Du Y. Water-solubility of chitosan and its antimicrobial activity. Carbohydrate Polymers. 2006; 63(3): 367-374. doi: 10.1016/j.carbpol.2005.09.023

Byrd JA, Anderson RC, Brewer RL, et al. Effect of lactic acid administration in the drinking water during preslaughter feed withdrawal on Salmonella and Campylobacter contamina-tion of broilers. Poult Sci. 2001; 80(3): 278-283. doi: 10.1093/ ps/80.3.278

Chaveerach P, Keuzenkamp DA, Lipman LJ, Van Knapen F. Effect of organic acids in drinking water for young broilers on Campylobacter infection, volatile fatty acid production, gut microflora and histological cell changes. Poult Sci. 2004; 83(3): 330-334. doi: 10.1093/ps/83.3.330

Rosenquist H, Nielsen NL, Sommer HM, Norrung B, Chris-tensen BB. Quantitative risk assessment of human campylobac-teriosis associated with thermophilic Campylobacter species in chickens. Int J Food Microbiol. 2003; 83(1): 87-103. doi: 10.1016/S0168-1605(02)00317-3

Olson CK, Ethelberg S, van Pelt W, Tauxe RV. Epidemiology of Campylobacter jejuni infections in industrialized nations. In: Nachamkin I, Szymanski C, Blaser M, eds. Campylobacter. 3rd ed. Washington: ASM Press; 2008: 163-189.

Sears A, Baker MG, Wilson N, et al. Marked campylobacte-riosis decline after interventions aimed at poultry, New Zealand. Emerg Infect Dis. 2001; 17(6): 1007-1015.

New Zealand Food Safety Authority. Campylobacter risk management strategy, 2013-2014. Website: http://www.food-safety.govt.nz/elibrary/industry/Campylobacter_Risk-Compre-hensive_Aimed.pdf 2013; Accessed September 23, 2015.

Published

2015-10-27

How to Cite

Hanna R. Arambel, Ann M. Donoghue, Komala Arsi, Abhinav Upadhyay, Ann Woo-Ming, Pamela J. Blore, … Dan J. Donoghue. (2015). Chitosan Supplementation Reduces Enteric Colonization of Campylobacter jejuni in Broiler Chickens and Down-Regulates Expression of Colonization Genes. Advances in Food Technology and Nutrition Sciences, 1(5), 104–111. https://doi.org/10.17140/aftnsoj-1-119