Global Advanced Research Journal of Microbiology (GARJM) ISSN: 2315-5116
June 2019 Vol. 8(4): pp. 063-071
Copyright © 2019 Global Advanced Research Journals

Full Length Research Paper

Genetic Characterization of Campylobacter jejuni Isolated from Boiler Flocks

H. Abo Al-Yazeed¹, Amani M. Marwan¹, Attia, A. M.², A. Samir¹, Ammar, A. M³

¹Department of Microbiology, Faculty of Veterinary Medicine, Cairo University, Cairo, Egypt.

² Department of Bacteriology, Animal Health Research Institute, Dokki, Giza.

³ Microbiology Department, Faculty of Veterinary Medicine, Zagazig University, Egypt.

Email: amanymarwan110@mail.com

Accepted 05 June, 2019

Abstract

Campylobacter  is a worldwide infection which has been estimated it as the most significant economic burden by EFSA and ECDC in 2016. It caused mainly by either Campylobacter  jejuni or Campylobacter coli. Poultry are a natural reservoir for Campylobacter species, although it is insignificant for poultry health and is a leading cause of gastroenteritis for humans in developed and developing countries. The disease is endemic in Egypt and is a major cause for diarrhea in children. Good understanding of epidemiology and surveillance of Campylobacter will help in elimination and prevention of it among animals and humans. Many molecular typing techniques used to track the source of infection and reduce Campylobacter infection rate. To achieve that, 290 samples were collected from broiler flocks and slaughter market from Cairo governorate, Egypt. Bacteriological and molecular identification have been implemented based on genus and species level for C. jejuni then phylogenetic tree analysis of flaA gene to correlate the genetic relatedness and emphasize that chicken is the major source of infection.

Keywords:  Campylobacter jejuni, genotyping, flaA gene, short variable region, chicken, Human.

REFERENCES

Achtman M, Wagner M (2008). Microbial diversity and the genetic nature of microbial species. Nat. Rev. Microbiol. 6, 431–440. [CrossRef] [PubMed].

Amar CF, East CL, Gray J, Iturriza-Gomara M, Maclure EA, McLauchlin J (2007). Detection by PCR of eight groups of enteric pathogens in 4,627 faecal samples: re-examination of the English case-control Infectious Intestinal Disease Study (1993–1996). Eur J Clin Microbiol Infect Dis. 26(5):311–23. Epub 2007/04/21. PMID: 17447091.

Asmaa G, Youseef I, Ibrahim A, Sayed SM, Mona MS (2017). Occurrence of Campylobacter species in chickens by multiplex polymerase chain reaction. Assiut Vet. Med. J. Vol. 63 No. 152 January 2017, 66-72.

Awadallah M, Ahmed H, El-Gedawy A, Saad AM (2014). Molecular identification of C. jejuni and C. coli in chicken and humans, at Zagazig, Egypt, with reference to the survival of C. jejuni in chicken meat at refrigeration and freezing temperatures. International Food Research Journal 21(5): 1801-1812.

Bang DD, Møller E, Nielsen F, Scheutz K, Pedersen K, Handberg M, Madsen (2003). PCR detection of seven virulence and toxin genes of Campylobacter jejuni and Campylobacter coli isolates from Danish pigs and cattle and cytolethal distending toxin production of the isolates. Journal of Applied Microbiology. 94, 1003–1014.

Banowary B, Dang VT, Sarker S, Connolly JH, Chenu J, Groves P (2015). Differentiation of Campylobacter jejuni and Campylobacter coli using Multiplex-PCR and High Resolution Melt Curve Analysis. PLoS ONE 10(9): e0138808.doi:10.1371/journal.pone.0138808.

Bartkowiak-Higgo AJ, Veary CM, Venter EH, Bosman AM (2006). A pilot study on post-evisceration contamination of broiler carcasses and ready-to-sell livers and intestines (mala) with Campylobacter jejuni and Campylobacter coli in a high-throughput South African poultry abattoir. J. S. Afr Vet Assoc.; 77(3): 114-9.

Bashor MP, Curtis PA, Keener KM, Sheldon BW, Kathariou S, Osborne JA (2004). Effects of carcass washers on Campylobacter contamination in large broiler processing plants. Poult. Sci. 83, 1232–1239. [CrossRef] [PubMed]

Best EL, Powell EJ, Swift C, Grant KA, Frost JA (2003). Applicability of a rapid duplex real-time PCR assay for speciation of Campylobacter jejuni and Campylobacter coli directly from culture plates. FEMS Microbiol Lett 229:237–241. doi:10.1016/S0378-1097(03)00845-0.

Bolton DJ (2015). Campylobacter virulence and survival factors. Food Microbiol. 48, 99–108. [CrossRef] [PubMed]

Burch D (2005). Avian vibrionic hepatitis in laying hens. Vet. Rec. 157:528.

Carron M, Alarcon P, Karani M, Muinde P, Akoko J, Onon J (2017). The broiler meat system in Nairobi, Kenya: Using a value chain framework to understand animal and product flows, governance and sanitary risks. Prev Vet Med [Internet]. 147(August):90±9. Available from: https://doi.org/10.1016/j.prevetmed.2017.08.013

Centers for Disease Control and Prevention (1996). Incidence and Trends of Infection with Pathogens Transmitted Commonly Through Food — Foodborne Diseases Active Surveillance Network, 10 U.S. Sites, 1996–2012. April 19, 2013 / 62(15);283-287.

Centers for Disease Control and Prevention (2017). Campylobacter. Available online: https://www.cdc.gov/foodsafety/diseases/campylobacter/ (accessed on 4 May 2017).

Chuma IS, Nonga HE, Mdegela RH, Kazwala RR (2016). Epidemiology and RAPD-PCR typing of thermophilic campylobacters from children under five years and chickens in Morogoro Municipality, Tanzania. BMC Infect Dis [Internet]. 2016;1±11. Available from: http://dx.doi.org/10.1186/s12879-016-2031-z

Colles FM, Maiden MC (2012). Campylobacter sequence typing databases: Applications and future prospects. Microbiology.158, 2695–2709. [CrossRef] [PubMed]

Dasti JI, Tareen AM, Lugert R, Zautner AE, Gross U (2010). Campylobacter jejuni: A brief overview on pathogenicity-associated factors and disease-mediating mechanism. International Journal of Medical Microbiology 300, 205–211.

Datta S, Niwa H, Itoh K (2003). Prevalence of 11 pathogenic genes of Campylobacter jejuni by PCR in strains isolated from humans, poultry meat and broiler and bovine faeces. Journal of Medical Microbiology. 52, 345–348 DOI 10.1099.

Denis M, Soumet C, Rivoal K, Ermel G, Blivet D, Salvat G, Colin P (1999). Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli. Lett Appl Microbiol 29, 406–410.

Dingle KE, Colles FM, Falush D, Maiden MC (2005). Sequence typing and comparison of population biology of Campylobacter coli and Campylobacter jejuni. J Clin Microbiol 43:340-7

ECDC (2013). European center for disease prevention and control (ECDC).

Eunju S, Lee Y (2009). Comparison of Three Different Methods for Campylobacter Isolation from Porcine Intestines. J. Microbiol. Biotechnol., 19(7), 647–650.

European Food Safety Authority [EFSA] (2010). Analysis of the baseline survey on the prevalence of Campylobacter in broiler batches and of Campylobacter and Salmonella on broiler carcasses in the EU, 2008. EFSA J. 8, 1503–1602.

European Food Safety Authority; European Centre for Disease Prevention and Control (2016). The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2015. EFSA J. 2016, 14. [CrossRef]

Fitzgerald C, Helsel LO, Nicholson MA, Olsen SJ, Swerdlow DL, Flahart R, Sexton J, Fields PI (2001). Evaluation of methods for subtyping Campylobacter jejuni during an outbreak involving a food handler. J. Clin. Microbiol., 39, 2386-2390.

Gerner-Smidt P, Hise K, Kincaid J, Hunter S, Rolando S, Hyytia-Trees E (2006). PulseNet USA: a five-year update. Foodborne Pathog Dis.  3(1):9–19. Epub 2006/04/11. doi: 10.1089/fpd.2006.3.9 PMID: 16602975.

Ghorashi SA, Bradbury JM, Ferguson-Noel NM, Noormohammadi AH (2013). Comparison of multiple genes and 16S-23S rRNA intergenic space region for their capacity in high resolution melt curve analysis to differentiate Mycoplasma gallisepticum vaccine strain ts-11 from field strains. Vet Microbiol. 167 (3–4):440–7. Epub 2013/11/19. doi: 10.1016/j.vetmic.2013.09.032 PMID: 24238667.

Giombelli A, Gloria MBA (2014). Prevalence of salmonella and campylobacter on broiler chickens from farm to slaughter and efficiency of methods to remove visible fecal contamination. J. Food Prot. 77:1851–1859. Available at http://openurl.ingenta.com/content/xref?genre=article&issn= 0362028X&volume=77&issue=11&spage=1851.

Gomes CN, Souza RA,  Passaglia J,  Duque SS, Medeiros MI, Falcão JP (2016). Genotyping of Campylobacter coli strains isolated in Brazil suggests possible contaminationamongst environmental, human, animal and food sources. J Med Microbiol. 65(1):80-90.

Gruntar, I., Biasizzo, M., Kušar, D., Pate, M., Ocepek, M. 2015. Campylobacter jejuni contamination of broiler carcasses: population dynamics and genetic profiles at slaughterhouse level. Food Microbiol. 50, 97–101. doi: 10.1016/j.fm. 2015.03.007.

Hald B, Skovgàrd H, Bang DD, Pedersen K, Dybdahl J, Jespersen JB, Madsen M (2004). Flies and Campylobacter infection of broiler flocks. Emerg. Infect. Dis.10, 1490-1492.

Hariharan H, Sharma S, Chikweto A, Matthew V, Deallie C (2009). Antimicrobial drug resistance as determined by the E-test in Campylobacter jejuni, C. coli, and C. lari isolates from the ceca of broiler and layer chickens in Grenada. Comp Immunol Microbiol Infect Dis 32, 21–28.

Harrington CS, Thomson-Carter FM, Carter PE (1997). Evidence for recombination in the flagellin locus of Campylobacter jejuni: implications for the flagellin gene typing scheme. J. Clin. Microbiol., 35, 2386-2392.

Haruna M, Sasaki Y, Murakami M, Ikeda A, Kusukawa M, Tsujiyama Y (2012). Prevalence and antimicrobial susceptibility of Campylobacter in broiler flocks in Japan. Zoonoses Public Health 59, 241–245. doi: 10.1111/j.1863-2378.2011.01441.x

Hassan A, El Fadaly, Ashraf MAB, Khaled A.,Abd El-Razik, Azza SM, Abuelnaga, Elgabry AE (2016). Incidence of zoonotic Campylobacter jejuni in fast meal meat, grill chickens and symptomatic Egyptians. Afr. J. Microbiol. Res. Vol. 10 (18), pp. 608-615.

Hiett KL, Stern NJ, Fedorka-Cray P, Cox NA, Musgrove MT, Ladely S (2002). Molecular Subtype Analyses of Campylobacter spp. from Arkansas and California Poultry Operations. Appl. Environ. Microbiol., 68, 6220-6236.

Ioannidis A, Nicolaou C, Legakis NJ, (2006).The first database comprised of flagellin gene (flaA) types of Campylobacter jejuni human clinical isolates from Greece.Eur J Epidemiol. 21: 823. https://doi.org/10.1007/s10654-006-9073-2.

ISO – International Standards Organization (2006). Microbiology of food and animal feeding stuffs – horizontal method for detection and enumeration of Campylobacter spp. – Part 1: detection method. ISO 10272–1:2006.

Jensen AN, Andersen MT, Dalsgaard A, Baggesen DL, Nielsen EM (2005). Development of real-time PCR and hybridization methods for detection and identification of thermophilic Campylobacter spp. in pig faecal samples. J Appl Microbiol 99, 292–300.

Khalifa NO, Jehan SA, Afify Nagwa S, Rabie (2013). Zoonotic and Molecular Characterizations of Campylobacter jejuni and Campylobacter coliisolated from beef cattle and children. Global Veterinaria 11 (5): 585-591.

Linton D, Gilbert M, Hitchen PG, Dell A, Morris HR, Wakarchuk WW, Gregson NA, Wren BW (2000). Phase variation of a beta-1,3-galactosyltransferase involved in generation of the ganglioside GM1-like oligosaccharide of Campylobacter jejuni. Mol Microbiol 37, 501–514.

Magana M, Chatzipanagiotou S, Burriel AR, Ioannidis A (2017). Inquiring into the Gaps of Campylobacter Surveillance Methods. Vet Sci. 4(3):36. Published 2017 Jul 19. doi:10.3390/vetsci4030036.

Mageto LM, Ombui JN, Mutua FK (2018). Prevalence and Risk Factors for Campylobacter Infection of Chicken in Peri-Urban Areas of Nairobi Kenya. J Dairy, Vet Anim Res. 7(1).

Man SM (2011). The clinical importance of emerging Campylobacter species. Nat Rev Gastroenterol Hepatol 8:669–685. http://dx.doi.org/10.1038/nrgastro.2011.191.

Meinersmann RJ, Helsel LO, Fields PI, Hiett KL (1997). Discrimination of Campylobacter jejuni isolates by flaA gene sequencing. J Clin Microbiol. 35(11):2810–4. Epub 1997/11/14. PMID: 9350739; PubMed Central PMCID: PMC230067.

Mostafa FA, Sylvia OA, Awad AI, Hanan AM (2018). Prevalence of Zoonotic Species of Campylobacter in Broiler Chicken and Humans in Assiut Governorate, Egypt. Appro Poult Dairy & Vet Sci. 3(4). APDV.000568.2018. DOI: 10.31031/APDV.2018.03.000568.

Nachamkin I, Bohachick K, Patton CM (1993). Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis. J Clin Microbiol. 31(6):1531–6. Epub 1993/06/01. PMID: 8100241; PubMed Central PMCID: PMC265573.

Nachamkin I, Ung H, Patton CM (1996). Analysis of HL and O serotypes of Campylobacter strains by the flagellin gene typing system. Journal of Clinical Microbiology. 34, 277–281.

Nayak R, Stewart TM, Nawaz MS (2005). PCR identification of Campylobacter coli and Campylobacter jejuni by partial sequencing of virulence genes. Mol. Cell Probes., 19: 187-193.

Ngulukun S, Oboegbulem S, Klein G (2016). Multilocus sequence typing of Campylobacter jejuni and Campylobacter coli isolates from poultry, cattle and humans in Nigeria. J Appl Microbiol. 121:561±568. https://doi.org/10.1111/jam.13185 PMID: 27206561

Nielsen EM, Engberg J, Fussing V, Petersen L, Brogren CH, On SLW (2000). Evaluation of phenotypic and genotypic methods for subtyping Campylobacter jejuni isolates from humans, poultry, and cattle. J. Clin. Microbiol.. 38, 3800-3810.

Nishimura M, Nukina M, Yuan JM, Shen BQ, Ma .JJ, Ohta M, Saida T, Uchiyama T (1996). PCR-based restriction fragment lengt polymorphism (RFLP) analysis and serotyping of Campylobacter jejuni isolates from diarrheic patients in China and Japan. FEMS Microbiol. Lett., 142, 133-138.

Nuijten PJ, van den Berg AJ, Formentini I, van der Zeijst BA, Jacobs AA (2000). DNA rearrangements in the flagellin locus of flaA mutant of Campylobacter jejuni during colonization of chicken ceca. Infect. Immun., 68: 7137 7140.

On SL, Jordan PJ (2003). Evaluation of 11 PCR assays for species-level identification of Campylobacter jejuni and Campylobacter coli. J Clin Microbiol. 41(1):330-6.

Parkar SFD, Sachdev D, deSouza N, Kamble A, Suresh G, Munot H, Hanagal D, Shouche Y, Kapadnis B (2013). Prevalence, seasonality and antibiotic susceptibility of thermophilic Campylobacter spp. in ceca and carcasses of poultry birds in the “live- bird market”. African Journal of Microbiological Research 7 (21): 2442-2453.

Pendleton S, Hanning I, Biswas D, Ricke SC (2013). Evaluation of whole-genome sequencing as a genotyping tool for Campylobacter jejuni in comparison with pulsed-field gel electrophoresis and flaA typing. Poultry Science, Volume 92, Issue 2, Pages 573–580, https://doi.org/10.3382/ps.2012-02695.

Persson S, Olsen KE (2005). Multiplex PCR for identification of Campylobacter coli and Campylobacter jejuni from pure cultures and directly on stool samples. J. Med. Microbiol., 54: 1043-1047.

Quinn TC (1995). DNA amplification assays: a new standard for diagnosis of Chlamydia trachomatis infections. Ann Acad Med Singapore. 24(4):627–33. Epub 1995/07/01. PMID: 8849200.

Ribot EM, Fitzgerald C, Kubota K, Swaminathan B, Barrett TJ (2001). Rapid pulsed-field gel electrophoresis protocol for subtyping of Campylobacter jejuni. J Clin Microbiol 39:1889-94.

Rosenquist H, Boysen L, Krogh AL, Jensen AN, Nauta M (2013). Campylobacter contamination and the relative risk of illness from organic broiler meat in comparison with conventional broiler meat. Int J Food Microbiol. 162(3):226–30. Epub 2013/03/05. doi: 10.1016/j.ijfoodmicro.2013.01.022 PMID:23454812.

Rozynek E, Dzierzanowska-Fangrat K, Jozwiak. P, Popowski J, Korsak D, Dzierzanowska D (2005). Prevalence of potential virulence markers in Polish Campylobacter jejuni and Campylobacter coli isolates obtained from hospitalized children and from chicken carcasses. J. Med. Microbiol., 54: 615 619.

Sails AD, Swaminathan B, Fields PI (2003). Utility of multilocus sequence typing as an epidemiological tool for investigation of outbreaks of gastroenteritis caused by Campylobacter jejuni. J. Clin. Microbiol. 41:4733– 4739.

Sambrook J, Russell WD (2001). Molecular Cloning: a Laboratory Manual, 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.

Sasaki Y, Maruyama N, Zou B, Haruna M, Kusukawa M, Murakami M (2013). Campylobacter cross-contamination of chicken products at an abattoir. Zoonoses Public Health 60, 134–140. doi: 10.1111/j.1863-2378.2012. 01509.x

Shi F, Chen YY, Wassenaar TM, Woods HW, Coloe PJ, Fry BN (2002). Development and application of a new scheme for typing Campylobacter jejuni and Campylobacter coli by PCR-based restriction fragment length polymorphism analysis. J. Clin.Microbiol.. 40, 1791-1797.

Shobo CO, Bester LA, Baijnath S, Somboro AM, Peer AKC, Essack SY (2016). Antibiotic resistance profiles of Campylobacter species in the South Africa private health care sector. J Infect Dev Ctries. 10:1214±1221. https://doi.org/10.3855/jidc.8165 PMID: 27886034

Stephens CP, On SL, Gibson JA (1998). An outbreak of infectious hepatitis in commercially reared ostriches associated with Campylobacter coli and Campylobacter jejuni. Vet. Microbiol. 61:183–190.

Stoyanchev TT (2004). Detection of Campylobacter using standard culture and PCR of 16S rRNA gene in freshly chilled poultry and poultry products in a slaughter house. Trakia Journal of Sciences. 2(3): 59-64.

USDA (2008). Foodborne Illness: What Consumers Need to Know. Available at:http://www.fsis.usda.gov/wps/portal/fsis/topics/food-safety-education/get-answers/food-safety-fact-sheets/foodborne-illness-and-disease/foodborne-illness-what-consumers-need-to-know/CT_Index

van Asselt ED, de Jong AE, de Jonge R, Nauta MJ (2008). Cross-contamination in the kitchen: estimation of transfer rates for cutting boards, hands and knives. J Appl Microbiol 2008 Nov;105(5):1392-401. doi: 10.1111/j.1365-2672.2008.03875.x.

Vashin I, Stoyanchev T, Ring Ch, Atanassova V (2009). Prevalence of Campylobacter spp in frozen poultry giblets at Bulgarian retail markets. Trakia Journal of Sciences, 7 (4): 55-57.

Vidal AB, Rodgers J, Arnold M, Clifton-Hadley F (2013). 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 60:412–425. doi:10.1111/zph.12009.

Wagenaar JA, French NP, Havelaar AH (2013). Preventing Campylobacter at the source: Why is it so difficult? Clin. Infect. Dis. 57, 1600–1606. [CrossRef] [PubMed]

Wang G, Clark CG, Taylor TM, Pucknell C, Barton C, Price L, Woodward DL, Rodgers FG (2002). Colony Multiplex PCR Assay for Identification and Differentiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subsp. Fetus. Journal of Clinical Microbiology, Vol. 40, No. 12.  p. 4744–4747.

Whiley H, van den Akker B, Giglio S, Bentham R (2013). The role of environmental reservoirs in human campylobacteriosis. Int J Environ Res Public Health. 10(11):5886–907. Epub 2013/11/13. doi: 10.3390/ijerph10115886 PMID: 24217177; PubMed Central PMCID: PMC3863877.

Wieczorek K (2010). Antimicrobial resistance and virulence markers of campylobacter jejuni and Campylobacter coli isolated from retail poultry meat in Poland. Bull. Vet. Inst. Pulawy. 54; 563-569.

Wieczorek K, Osek J (2005). Multiplex PCR assays for simultaneous identification of Campylobacter jejuni and Campylobacter coli. Medycyna weterynaryjna 61(7):797-799.

Wingstrand A, Neimann J, Engberg J, Nielsen EM, Gerner-Smidt P, Wegener HC (2006). Fresh chicken as main risk factor for campylobacteriosis, Denmark. Emerg Infect Dis. 12(2):280–5. Epub 2006/02/24. doi: 10.3201/eid1202.050936 PMID: 16494755; PubMed Central PMCID: PMC3373097.

Zhang X, Tang M, Zhou Q, Zhang J, Yang X, Gao Y (2018). Prevalence and Characteristics of Campylobacter Throughout the Slaughter Process of Different Broiler Batches. Front. Microbiol. 9: 2092.doi: 10.3389/fmicb.2018.02092.

Ziprin RL, CR, Young JA, Byrd LH, Stanker ME, Hume SA, Gray BJ, Kim Konkel ME (2001). Role of Campylobacter jejuni potential virulence genes in cecal colonization. Avian Dis., 45: 549 557.

Zumbaco-Guti´errez L, Ar´evalo-Madrigal A, Donado-Godoy M, Romero-Z´uniga J (2014). DIAGNOSTICO MOLECULAR DE Campylobacter EN LA Cadena. Agron. Mesoam. 25:357–363. Available at http://www.mag.go.cr/rev meso/v25n02 357.pdf.