Volume 2, 2012

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Extensive research and field trials have identified a wide range of management and intervention measures for the reduction of human enteropathogens from poultry. Vaccination of poultry has been shown to reduce carriage of Salmonella into the processing plant.
Introduction ontrol of foodborne pathogens remains a major challenge for the poultry industry and of critical importance to public health. Efforts to minimize or eliminate foodborne pathogens, such Sandra Kelly-Aehle, as Salmonella, CampyloTechnical Manager bacter and Listeria, must include preharvest and postharvest strategies. Vaccines developed for poultry have shown to be effective in reducing Salmonella in the environment and in reducing vertical transmission of infection. Salmonella vaccination programs, along with other control measures, have been implemented successfully in providing a last line of defense for the bird against infection by field strains. To date, no other vaccines are commercially available for poultry for the prevention of human bacterial enteropathogens such as Campylobacter and Listeria. Vaccination with live or killed vaccines is designed to stimulate maximal immunity and provide a first line of defense that directly increases the level of resistance to Salmonella infection in the bird. As a result, a reduction in intestinal colonization by salmonellae results in reduced fecal shedding into the environment. A reduction in intestinal colonization also prevents systemic infection that inherently results in infection of the internal organs and reproductive tissues. The USDA FSIS recommends the use of vaccination programs for poultry as one of several interventions to reduce foodborne pathogens (9). Live and inactivated vaccines to reduce colonization of the gut and reproductive

A LO H M A N N A N I M A L H E A LT H N E W S B R I E F

Current advances in immunization of poultry against foodborne pathogens

organs by Salmonella are commercially available and when used together in a Salmonella prevention program have been demonstrated both by controlled and field studies to effectively reduce vertical transfer of Salmonella carriage and to confer protective immunity to progeny of vaccinated breeders (1, 7, 14, 26). Studies conducted by FDA showed AviPro MeganVac1 generated significant cell-mediated immunity that effectively cleared wild-type S. Enteritidis from vaccinated birds (2). Studies in hens undergoing molt have shown significant protection against challenge with wild-type S. Enteritidis after one spray application of a live Salmonella vaccine prior to feed withdrawal (15). Hassan and Curtiss (13) demonstrated that vaccination of chickens with a live S. Typhimurium vaccine provided significant cross-protection to challenge with several serotypes in the B, D and E Salmonella serogroups. Campylobacter remains a challenge for the broiler industry. Risk factors for the occurrence of Campylobacter were identified in broiler ocks as age and ock size (16), animals in the vicinity of the broiler house, livestock other than chickens on the farm, a down period of less than 14 days, dividing the ocks for slaughter (12) and the practice of batch depletion (11). Stern et al (22) reported enhanced on-farm biosecurity practices and freezing of carcasses from Campylobacter-positive ocks contributed to the significant drop in poultry-associated campylobacteriosis in Iceland. Government regulations and implementation of stringent biosecurity practices in primary production were effective in preventing Campylobacter infection of ocks and directly correlated to a 74% reduction in campylobacteriosis attributed to poultry in

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Current advances in immunization of poultry against foodborne pathogens, p.1

Notes from the CEO, p.4

New Zealand (20). Several experimental approaches, like the reduction of colonization by competitive exclusion, bacteriocins, and application of bacteriophage, are currently under investigation for their effectiveness in reducing Campylobacter in primary production (10, 21, 23). Although killed whole-cell Campylobacter vaccines have had limited success (6), Wyszynska et al (25) reported more than a 6-log reduction of Campylobacter jejuni after challenge of birds vaccinated with a live recombinant cya crp Salmonella vaccine strain synthesizing the highly conserved immunodominant CjaA protein. Buckley and coworkers (4) found that an attenuated aroA Salmonella Typhimurium strain synthesizing CjaA fused to TetC reduced the challenge C. jejuni organisms in the ceca by 1.4 log in chickens. More recently, Layton et al (18) found that oral vaccination with aroA htrA S. Enteritidis synthesizing CjaD, an outer membrane protein, reduced C. jejuni colonization in ileal mucosal samples by 4.8 log to undetectable levels. The Centers for Disease Control demonstrated a direct epidemiological link between human listeriosis and the consumption of undercooked poultry (19). Listeria is widely distributed in nature (24) and it would be expected that poultry could be exposed during live production. However, the development of poultry vaccines against Listeria monocytogenes would not be economical due to the infrequent and low isolation of the organism from live birds (3, 8). The antimicrobial and protective mechanism of probiotic bacteria against Listeria has been investigated. Corr et al (5) demonstrated that bacteriocinsecreting Lactobacillus salivarius produced a consistent reduction of 1-2 logs in internal organs of mice challenged with over 109 CFU of L. monocytogenes. More interesting was evidence that supported direct antagonism as the mode of action involved in protection against L. monocytogenes as early as 30 minutes after administration of the engineered L. salivarius. Recent studies by Koo and coworkers (17) showed that Listeria adhesion protein (LAP) synthesized by recombinant probiotic Lactobacillus paracasei was able to interact and attach to heat shock protein on the surface of intestinal cells just as Listeria would. The L. paracasei physically crowded out Listeria organisms and decreased the

number of Listeria cells that passed through intestinal cells by 46%. The direct human use of bacteriophage was approved in 2006 by FDA when a phage preparation targeted against L. monocytogenes was allowed for spraying meat, thus opening the way for other phages to be recognized as having GRAS (generally recognized as safe) status (ListShield, http://www. Intralytix.com). Extensive research and field trials have identified a wide range of management and intervention measures for the reduction of human enteropathogens from poultry. Vaccination of poultry has been shown to reduce carriage of Salmonella into the processing plant. While many strategies for reducing foodborne pathogens have been proven efficacious in laboratory research, the conduct of true field trials under commercial settings requires continued research. References 1. Atterbury, R.J., J.J. Carrique-Mas, R.H. Davies and V.M. Allen. Salmonella colonization of laying hens following vaccination with killed and live attenuated commercial Salmonella vaccines. Vet Rec 165:493-496. 2009. 2. Babu, U., R.A. Dalloul, M. Okamura, H.S. Lillehoj, H. Xie, R.B. Raybourne, D. Gaines and R.A. Heckert. Salmonella enteritidis clearance and immune responses in chickens following Salmonella vaccination and challenge. Vet Immunol Immunopathol 101:251-257. 2004. 3. Berrang, M.E., C.E. Lyon, D.P. Smith and J.K. Northcutt. Incidence of Listeria monocytogenes on pre-scald and post-chill chicken. J. Appl. Poultry Res. 9:546-550. 2000. 4. Buckley, A.M., J. Wang, D.L. Hudson, A.J. Grant, M.A. Jones, D.J. Maskell and M.P. Stevens. Evaluation of live-attenuated Salmonella vaccines expressing Campylobacter antigens for control of C. jejuni in poultry. Vaccine 28:10941105. 2010. 5. Corr, S.C., Y. Li, C.U. Riedel, P.W. OToole, C. Hill and C.G.M. Gahan. Bacteriocin production as a mechanism for the anti-infective activity of Lactobacillus salivarius UCC118. PNAS 104:7617-7621. 2007.

6. de Zoete, M.R., J.P.M. van Putten and J.A. Wagenaar. Vaccination of chickens against Campylobacter. Vaccine 25:5548-5557. 2007. 7. Dorea, F., D.J. Cole, C. Hofacre, K. Zamperini, D. Mathis, M.P. Doyle, M.D. Lee and J.J. Maurer. Effect of Salmonella vaccination of chicken breeders on reducing carcass contamination of broiler chickens in integrated poultry operations. Appl. Environ. Microbiol. 76:7820-7825. 2010. 8. Fenlon, D.R., J. Wilson and W. Donachie. The incidence and level of Listeria monocytogenes contamination of food sources at primary production and initial processing. J. Appl. Bacteriol. 81:641-650. 1996. 9. FSIS, USDA. Compliance Guideline for Controlling Salmonella and Campylobacter in Poultry. Third Edition. 2010. Accessed: http://www.fsis.usda.gov/PDF/Compliance_Guide_Controling_Salmonella_ Campylobacter_Poultry_0510.pdf 10. Goode, D., V.M. Allen and P.A. Barrow. Reduction of experimental Salmonella and Campylobacter contamination of chicken skin by application of lytic bacteriophages. Appl. Environ. Microbiol. 69:5032-5036. 2003. 11. Hald, B., E. Rattenborg and M. Madsen. Role of batch depletion of broiler houses on the occurrence of Campylobacter sp. In chicken ocks. Lett. Appl. Microbiol. 32:253-256. 2001. 12. Hald, B., A. Wedderkopp, and M. Madsen. Thermophilic Campylobacter spp. in Danish broiler production: a cross sectional survey and a retrospective analysis of risk factors for occurrence in broiler ocks. Avian Pathol. 29:123-131. 2000. 13. Hassan, J.O. and R. Curtiss III. Development and evaluation of an experimental vaccination program using a live avirulent Salmonella typhimurium strain to protect immunized chickens against challenge with homologous and heterologous Salmonella serotypes. Infect. Immun. 62:5519-5527. 1994.

14. Hassan, J.O. and R. Curtiss III. Effect of vaccination of hens with an avirulent strain of Salmonella typhimurium on immunity of progeny challenged with wildtype Salmonella strains. Infect. Immun. 64:938-944. 1996. 15. Holt, P.S., R.K. Gast, and S. Kelly-Aehle. Use of a live attenuated Salmonella typhimurium vaccine to protect hens against Salmonella enteritidis infection while undergoing molt. Avian Dis. 47:656661. 2003. 16. Keener, K.M., M.P. Bashor, P.A. Curtis, B.W. Sheldon and S. Kathario. Comprehensive review of Campylobacter and poultry processing. Comprehensive Reviews in Food Science and Food Safety Vol. 3, 105-106. 2004. 17. Koo, O.K., M.A.R. Amalaradjou and A.K. Bhunia. Recombinant probiotic expressing Listeria adhesion protein attenuates Listeria monocytogenes virulence in vitro. PLoS ONE 7(1): e29277. doi:10.1371/journal.pone.0029277. 2012. 18. Layton, S.L., M.J. Morgan, K. Cole, Y.M. Kwon, D.J. Donoghue, B.M. Hargis and N.R. Pumford. Evaluation of Salmonella-vectored Campylobacter peptide epitopes for reduction of Campylobacter jejuni in broiler chickens. Clin. Vac. Immunol. 18:449-454. 2011. 19. Schwartz, B., C.A. Ciesielski, C.V. Broome, S. Gaventa, G.R. Brown, B.G. Gellin, A.W. Hightower and L. Mascola. Association of sporadic listeriosis with consumption of uncooked hot dogs and undercooked chicken. Lancet ii: 779-782. 1988. 20. Sears, A. M.G. Baker, N. Wilson, J. Marshall, P. Muellner, D.M. Campbell, R.J. Lake and N.P. French. Marked Campylobacteriosis decline after interventions aimed at

poultry, New Zealand. Emerg Infect Dis 17:1007-1015. 2011. 21. Stern, N.J., N.A. Cox, J.S. Bailey, M.E. Berrang and M.T. Musgrove. Comparison of mucosal competitive exclusion and competitive exclusion treatment to reduce Salmonella and Campylobacter spp. colonization in broiler chickens. Poult. Sci., 80:156-160. 2001. 22. Stern, N.J., K.L. Hiett, G.A. Alfredsson, K.G. Kristinsson, J. Reiersen, H. Hardardottir, H. Briem, E. Gunnarsson, F. Georgsson, R. Lowman, E. Berndtson, A.M. Lammerding, G.M. Paoli, and M.T. Musgrove. Campylobacter spp. in Icelandic poultry operations and human disease. Epidemiol. Infect. 130:23-32. 2003. 23. Svetoch, E.A., B.V. Eruslanov, V.V. Perelygin E.V. Mitsevich, I.P. Mitsevich, V.N. Borzenkov, V.P. Levchuk, O.E. Svetoch, Y.N. Kovalev, Y.G. Stepanshin, G.R. Siragusa, B.S. Seal and N.J. Stern. Diverse antimicrobial killing by Enterococcus faecium E 50-52 bacteriocin. J. Agric. Food Chem. 56:19421948. 2008. 24. Weis, J. and J.P.R. Seeliger. Incidence of Listeria monocytogenes in nature. Appl. Microbiol. 30:20-32. 1975. 25. Wyszynska, A., A. Raczko, M. Lis, E.K. Jagusztyn-Krynicka. Oral immunization of chickens with avirulent Salmonella vaccine strain carrying C. jejuni 72Dz/92 cjaA gene elicits specific humoral immune response associated with protection against challenge with wild-type Campylobacter. Vaccine 22:1379-1389. 2004. 26. Young, S.D., O. Olusanya, K.H. Jones, T. Liu, K.A. Liljebjelke and C.L. Hofacre. Salmonella incidence in broilers from breeders vaccinated with live and killed Salmonella. J. Appl. Poult. Res. 16:521-528. 2007.

Notes from the CEO

The level of sophistication required for successful poultry production has significantly increased over the past decade. Animal care and husbandry are now focused squarely on the grower, and the challenges are many. Consumers have become more educated about their food sources, and while they are sometimes misinformed about commercial poultry production, it is our responsibility to provide prodDave Zacek ucts that meet their health and safety CEO, Lohmann Animal Health expectations. Lohmann Animal Health International is a pioneer in both live and killed vaccines to control Salmonella in poultry. We are the world leader in live and killed Salmonella vaccines and programs for their use. Our live Salmonella vaccines are patented and both are first to be licensed by USDA with AviPro Megan Vac 1 and EU GMP for AviPro Salmonella Vac E. These vaccines have both shown to be effective control tools for prevention of Salmonella contamination. In addition, Lohmann Animal Health International provides inactivated Salmonella with and without ND+IB. Finally, for special situations, we lead in autogenous inactivated Salmonella vaccines made from strains from your operation. The author of this issue of Avian Insight has deep expertise and is available as a Consultant on Salmonella control approaches through our fee-based Lohmann Solutions. As always, we want to be your poultry health partner. Contact us for details on how we can help.

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