Molecular Detection of Campylobacter spp in Day-Old Chick Demonstrate Vertical Transmission in Poultry Production
Campylobacteriosis is the most common cause of foodborne gastrointestinal illness in the industrialized world, and poultry is considered the main source. While horizontal transmission is a route clearly linked to the spread of Campylobacter at farm level, few studies support the notion of vertical transmission. Currently, epidemiological research indicates that newly hatched chicks appear to be free of Campylobacter. Thus, we carried out the present study to investigate the occurrence of Campylobacter in day-old chicks using molecular methods to examine vertical transmission in poultry production. A total of 12 broiler flocks were monitored from the time of housing day-old chicks (day 1) and at the end of the rearing period (day 42). Samples were culture according with official method ISO 10272:2006 and analyzed using reverse transcription quantitative real-time PCR method. Our results revealed that no evidence of Campylobacter was found in the day-old chicks by bacterial culture method. Nevertheless, 4 flocks out of 12 were found to be positive by the molecular method. Real-time PCR identification revealed that C. coli was detected in all 4 flocks, while C. jejuni was identified in 3 flocks. No presence of Campylobacter spp. was observed in the environmental samples. These results reflect the evidence for vertical transmission of Campylobacter spp. While studies do not definitively rule out the detection problems and an accepted standard method will be developed for the detection and isolation of Campylobacter spp. at farm level, no standard measure may be successfully implemented in broiler production and therefore, from a public health point of view, strategies to reduce the number of human campylobacteriosis cases will not be efficient.
Poultry, Food Safety, Vertical Transmission, qPCR
Centers for Disease Control and Prevention (CDC). Vital signs: incidence and trends of infection with pathogens transmitted commonly through food – foodborne disease active surveillance network, 10 U.S. sites, 1996–2010. MMWR Morb Mortal Wkly Rep 2011;60:749-755.
EFSA (European Food Safety Authority). The European Union Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and Food-borne Outbreaks in 2012. EFSA Journal 2014;12:3547.
Scallan E, Hoekstra RM, Angulo FJ, Tauxe RV, Widdowson MA, Roy SL, Jones JL, Griffin PM. Foodborne illness acquired in the United States--major pathogens. Emerg. Infect. Dis. 2011;17:7-15.
Jacobs-Reitsma WF. Campylobacter in the food supply, in: Nachamkin, I., Blaser, M.J. (Eds.), Campylobacter, 2nd Edition. ASM Press, Washington, DC, 2000;467-481.
Corry JE, Atabay HI. Poultry as a source of Campylobacter and related organisms. J. Appl. Microbiol. 2001;90:96S–114S.
Cox NA, Richardson LJ, Maurer JJ, Berrang ME, Fedorka-Cray PJ, Buhr RJ, Byrd JA, Lee MD, Hofacre CL, O'Kane PM, Lammerding AM, Clark AG, Thayer SG, Doyle MP. Evidence for horizontal and vertical transmission in Campylobacter passage from hen to her progeny. J. Food. Prot. 2012;75:1896-1902.
Sahin O, Morishita TY, Zhang Q. Campylobacter colonization in poultry: sources of infection and modes of transmission. Anim. Health. Res. Rev. 2002;3:95-105.
Agunos A, Waddell L, Léger D, Taboada E. A systematic review characterizing on-farm sources of Campylobacter spp. for broiler chickens. PLoS One 2014;9:e104905.
Vidal AB, Rodgers J, Arnold M, Clifton-Hadley F. Comparison of different sampling strategies and laboratory methods for the detection of C. jejuni and C. coli from broiler flocks at primary production. Zoonoses Public Health 2013;60:412-
Gharst G, Oyarzabal OA, Hussain SK. Review of current methodologies to isolate and identify Campylobacter spp. from foods. J. Microbiol. Methods. 2013;95:84-92.
Ugarte-Ruiz M, Gómez-Barrero S, Porrero MC, Alvarez J, García M, Comerón MC, Wassenaar TM, Domínguez L. Evaluation of four protocols for the detection and isolation of thermophilic Campylobacter from different matrices. J. Appl. Microbiol. 2012;113:200-8.
Debretsion A, Habtemariam T, Wilson S, Nganwa D, Yehualaeshet T. Real-time PCR assay for rapid detection and quantification of Campylobacter jejuni on chicken rinses from poultry processing plant. Mol Cell Probes. 2007;21:177-81.
Botteldoorn N, Van Coillie E, Piessens V, Rasschaert G, Debruyne L, Heyndrickx M, Herman L, Messens W. Quantification of Campylobacter spp. in chicken carcass rinse by real-time PCR. J Appl Microbiol. 2008;105:1909-18.
Melero B, Cocolin L, Rantsiou K, Jaime I, Rovira J. Comparison between conventional and qPCR methods for enumerating Campylobacter jejuni in a poultry processing plant. Food Microbiol. 2011;28:1353-8.
Bui XT, Wolff A, Madsen M, Bang DD. Reverse transcriptase real-time PCR for detection and quantification of viable Campylobacter jejuni directly from poultry faecal samples. Res. Microbiol. 2012;163:64-72.
Bui XT, Wolff A, Madsen M, Bang DD. Fate and Survival of Campylobacter coli in Swine Manure at Various Temperatures. Front. Microbiol. 2011;2:262.
Anonymous. Microbiology of food and animal feeding stuffs—polymerase chain reaction (PCR) for the detection of food borne pathogens—requirements for amplification and detection for qualitative methods. ISO 20838. International Organization for Standardization, Geneva, Switzerland. 2006.
Newell DG, Fearnley C. Sources of Campylobacter colonization in broiler chickens. Appl. Environ. Microbiol. 2003;69:4343-51.
Jacobs-Reitsma WF, van de Giessen AE, Bolder NM, Mulder RW. Epidemiology of Campylobacter spp. at two Dutch broiler farms. Epidemiol. Infect 1995;114:413-21.
Evans SJ, Sayers AR. A longitudinal study of Campylobacter infection of broiler flocks in Great Britain. Prev. Vet. Med. 2000;46:209-223.
Newell DG, Wagenaar JA. Poultry infections and their control. In: Campylobacter, 2nded. I. Nachamkin, ed. J.J. Blaser. ASM Press, Washington, DC. 2000;497-510.
Shreeve JE, Toszeghy M, Pattison M, Newell DG. Sequential spread of Campylobacter infection in a multipen broiler house. Avian Dis. 2000;44:983-8.
Rivoal K, Ragimbeau C, Salvat G, Colin P, Ermel G. Genomic diversity of Campylobacter coli and Campylobacter jejuni isolates recovered from free-range broiler farms and comparison with isolates of various origins. Appl. Environ. Microbiol. 2005;71:6216-6227.
Bull SA, Allen VM, Domingue G, Jorgensen F, Frost JA, Ure R, Whyte R, Tinker D, Corry JE, Gillard-King J, Humphrey TJ. Sources of Campylobacter spp. colonizing housed broiler flocks during rearing. Appl. Environ. Microbiol. 2006;72:645-652.
Messens W, Herman L, De Zutter L, Heyndrickx M. Multiple typing for the epidemiological study of contamination of broilers with thermotolerant Campylobacter. Vet. Microbiol. 2009;138:120-131.
Allen VM, Ridley AM, Harris JA, Newell DG, Powell L. Influence of production system on the rate of onset of Campylobacter colonization in chicken flocks reared extensively in the United Kingdom. Br. Poult. Sci. 2011;52:30-39.
Jacobs-Reitsma WF. Aspects of epidemiology of Campylobacter in poultry. Vet Q. 1997;19:113-7.
Adkin A, Hartnett E, Jordan L, Newell D, Davison H. Use of a systematic review to assist the development of Campylobacter control strategies in broilers. J. Appl. Microbiol. 2006;100: 306-315.
Van Gerwe TJ, Bouma, A, Jacobs-Reitsma WF, van den Broek J., Klinkenberg D, Stegeman JA, Heesterbeek JA. Quantifying transmission of Campylobacter spp. among broilers. Appl. Environ. Microbiol. 2005;71:5765-70.
Lin J. Novel approaches for Campylobacter control in poultry. Foodborne Pathog. Dis. 2009;6:755-65.
Flekna G, Schneeweiss W, Smulders FJ, Wagner M, Hein I. Real-time PCR method with statistical analysis to compare the potential of DNA isolation methods to remove PCR inhibitors from samples for diagnostic PCR. Mol. Cell. Probes. 2007;21:282-287.
Humphrey T, O’Brien S, Madsen M. Campylobacter as zoonotic pathogens: a food production perspective. Int J Food Microbiol 2007;117, 237–257.
Chuma T, Yamada T, Yano K, Okamoto K, Yugi H. A survey of Campylobacter jejuni in broilers from assignment to slaughter using DNA-DNA 360 hybridization. J. Vet. Med. Sci. 1994;56:697-700.
Chuma T, Yano K, Omori H, Okamoto K, Yugi H. Direct detection of Campylobacter jejuni in chicken cecal contents by PCR. J. Vet. Med. Sci. 1997;59:85-87.
Chuma T, Makino K, Okamoto K, Yugi H. Analysis of distribution of Campylobacter jejuni and Campylobacter coli in broilers by using restriction fragment length polymorphism of flagellin gene. J. Vet. Med. Sci. 1997;59:1011-1015.
Kruger NJ, Buhler C, Iwobi AN, Huber I, Ellerbroek L, Appel B, Stingl K. “Limits of control”–crucial parameters for a reliable quantification of viable Campylobacter by real-time PCR. PLoS One 2014;9:e88108.
Jokinen CC, Koot JM, Carrillo CD, Gannon VP, Jardine CM, Mutschall SK, Topp E, Taboada EN. An enhanced technique combining pre-enrichment and passive filtration increases the isolation efficiency of Campylobacterjejuni and Campylobacter coli from water and animal fecal samples. J Microbiol Methods 2012;91:506-421 513.
Idris U, Lu J, Maier M, Sanchez S, Hofacre CL, Harmon BG, Maurer JJ, Lee MD. Dissemination of fluoroquinolone-resistant Campylobacter spp. within an integrated commercial poultry production system. Appl. Environ. Microbiol. 2006;72:3441-3447.
Hiett KL, Cox NA, Rothrock MJJr. Polymerase chain reaction detection of naturally occurring Campylobacter in commercial broiler chicken embryos. Poult. Sci. 2013;92:1134-1137.