Detection of cytolethal distending toxin (cdt) Genes in Campylobacter jejuni and Campylobacter coli isolated from the intestinal of commercial broiler chickens, turkey and quail of Iran

Authors

1 Department of Avian Diseases, Faculty of Veterinary Medicine, University of Tehran, Iran

2 Razi Vaccine and Serum Research Institute, Karaj, Iran

3 Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, University of Tehran, Iran

4 Department of Clinical Pathology, Faculty of Veterinary Medicine, University of Tehran, Iran

5 Department of Microbiology, Faculty of Veterinary Medicine, University of Tehran, Iran

Abstract

Background: Campylobacter jejuni and Campylobacter coli are zoonotic bacteria which are frequently associated with human diarrhea. Sharing of the cytolethal distending toxin (cdt) genes in Campylobacter is common and is considered species specific. OBJECTIVES: In this study we focused on detecting the presence of cdt gene in C. jejuni and C. coli isolated from broilers, turkeys and quails of Iran. METHODS: Cecal samples were randomly collected from 240 broiler chickens, 100 meat type turkeys and 100 quails after slaughtering. We used PCR as a method for detecting cdt genes. RESULTS: In broilers, 93% of 58 C. jejuni positive samples possessed cdt gene and in all cases the three different subunits of cdt genes were present. However, only 56% of 14 C. coli isolates in broilers had contained cdt genes, while one fourth having all three subunits present. In turkeys, around 65% of 34 C. jejuni positive samples had cdt gene present with 38% possessing all three subunits of cdt genes. But all 5 C. coli isolates had all three subunits cdt gene. In quails, 67% of 30 C. jejuni positive samples were identified by cdt gene, 20% of those possessed all three gene subunits. On the other hand, all 28 C. coli isolates of quails had cdt gene present while 36% of those held all three gene subunits. CONCLUSIONS: Our data is indicating the isolation, culture and cdt PCR amplification approaches in this study seemed to be efficient. However, the presence of different variation of Campylobacter cdt gene types in our sample isolates signifies the necessity of further functional gene studies to elucidate which gene type combinations result in encoding effective toxins.
 

Keywords


Asakura, M., Samosornsuk, W., Hinenoya, A. (2008) Development of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection and identification of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus. FEMS Immun. Med Microbiol. 52: 260-266.
Bang, D.D., Scheutz, F., Ahrens, P., Pedersen, K., Blom, J., Madsen, M. (2001) Prevalence of  cytolethal distending toxin (cdt) genes and CDT production in Campylobacter  spp. Isolated from Danish broilers. J Med Microbiol. 50: 1087-1094.
Blaser, M.J., Taylor, D.N., Feldman, R.A. (1983) Epidemiology of Campylobacter jejuni infection in humans. Epidemiol Rev. 5: 157– 176.
Di Giannatale, E. ,Prencipe, V., Colangeli, P., Alessiani, A., Barco, L., Staffolani, M., Tagliabue, S., Grattalora, C., Cerrone, A., Costa, A., Pisanu, M., Santucci, u., Iannitto, G., Migliorati, G. (2012) Prevalence of thermotolerant campylobacter in broiler flocks and broiler carcasses in Italy. Vet Ital. 46: 415-423.
Dipineto, L., Gargiulo, A., Russo, T.P., De Luca Bossa, L.M., Borrelli, L., Menna, L.F., Fioretti, A. (2010) Campylobacter jejuni, Campyloacter coli and cytolethal distending toxin genes in laying hens. Avian Dis. 55: 103.
Ehsannejad, F., Sheikholmolooki, A., Hassanzadeh, M., Shojaei Kavan, R., Soltani, M. (2015) Detection of cytolethal distending toxin (cdt) genes of Campylobacter Jejuni and Coli in fecal samples of pet birds in Iran. Iran J Vet Med. 9: 49- 56.
Findik, A., Ica, T., Emek Onuk, E., Percin, D., Kevenk, T.O., Ciftci, A. (2011) Molecular typing and cdt genes prevalence of Campylobacter jejuni isolates from various sources. Trop Anim Health. Prod. 43: 711–719.
Gonzales, I., Grant, K.A., Richardson, P.T., Park, S.F., Collins, M.D. (1997) Specific identification of the enteropathogens Campylobacter jejuni and Campylobacter coli by using a PCR test based on the ceuE gene encoding a putative virulence determinant. J Clin Microbiol. 35: 759–63.
Konkel, M.E., Garvis, S.G., Tipton, S.L., Anderson, Jr.D.E., Cieplak, Jr.W. (1997) Identification and molecular cloning of a gene encoding a fibronectin-binding protein (CadF) from Campylobacter jejuni. Mol Microbiol. 24: 953–63.
Konkel, M.E., Gray, S.A., Kim, B.J., Garvis, S.G., Yoon, J. (1999) Identification of the enteropathogens Campylobacter jejuni and Campylobacter coli based on the cadF virulence gene and its products. Clin Microbiol. 37: 510–7.
Lara-Tejero, M., Galàn, J.E. (2001) Cdt A, Cdt B, and Cdt C form a tripartite complex that is required for cytolethal distending toxin activity. Infect Immun. 69: 4358–65.
Lara-Tejero, M., Galàn J.E. (2002) Cytolethal distending toxin: limited damage as a strategy to modulate cellular functions. Trends Microbiol. 10: 147–52.
Linton, D., Owen, R.J., Stanley, J. (1996) Rapid identification by PCR of the genus Campylobacter and of five Campylobacter species enteropathogenic for man and animal. Res Microbiol. 147: 707-718.
Mirzaie, S., Hassanzadeh, M., Bashashati, M., Barrin, A. (2011) Campylobacter occurrence and antimicrobial resistance in samples from ceca of commercial turkeys and quails in Tehran, Iran. Int Res J Microbiol. 2: 338-342.
Nesic, D., Stebbins, C.E. (2005) Mechanisms of assembly and cellular interactions for the bacterial genotoxin CDT. PLoS Pathog. 1: 214–24.
On, S.L.W., Holmes, B. (1991) Reproducibility of tolerance tests that are useful in identification of campylobacteria. J Clin Microbiol. 29: 1785-1788.
On, S.L.W., Holmes, B. (1992) Assessment of enzyme detection tests useful in identification of campylobacteria. J Clin Microbiol. 30: 746-749.
Park, S.F., Richardson, P.T. (1995) Molecular characterization of a Campylobacter jejuni lipoprotein with homology to periplasmic siderophore binding proteins. J Bacteriol. 177: 2259–64.
Pickett, C.L., Whitehouse, C.A. (1999) The cytolethal distending toxin family. Trends Microbiol. 7: 292–7.
Rahimi, E. (2013) Campylobacter spp. contamination of chicken meat and by-products in Shahrekord, Iran. Iran Vet J. 9: 30-36.
Rahimi, E., Tajbakhsh, E. (2008) Prevalence of Campylobacter species in poultry meat in the Esfahan city, Iran. Bulg J Vet Med. 11: 257-262.
Rahimi, E., Ameri, M. (2011) Antimicrobial resistance patterns of Campylobacter spp. isolated from raw chicken, turkey, quail, partridge, and ostrich meat in Iran. Food Control. 22: 1165-70.
Son, I.,  Englen, M.D., Berrang, M.E., Fedorka-Cray, P.J., Harrison, M.A. (2007) Prevalance of Arcobacter and Campylobacter on broiler carcasses during processing. Inter J Food Microbiol. 113: 16-22.
Stern, N.J., Line, J.E. (1992) Comparison of three methods for recovery of Campylobacter spp. from broiler carcasses. J Food Prot. 55: 663–666.
Taremi, M., Soltan Dallal, M.M., Gachkar, L., Moez Ardalan, S., Zolfagharian, K., Zali, M.R. (2006) Prevalence and antimicrobial resistance of Campylobacter isolated from retail raw chicken and beef meat, Tehran, Iran. Int J Food Microbiol, 108: 401-403.
Van Deun, K., Haesebrouck, F., Heyndrickx, M., Favoreel, H., Dewulf, J., Ceelen, L., Dumez, L., Messens, W., Leleu, S., Van Immerseel, F.  Ducatelle, R. and Pasmans, F. (2007). Virulence properties of Campylobacter jejuni isolates of poultry and human origin. J Med Microbiol. 56: 1284–1289.
Wallace, J.S., Stanley, K.N., Currie, J.E., Diggle, P.J., Jones, K. (1997) Seasonality of thermophilic Campylobacter populations in chickens. J Appl Microbiol. 82: 219–224.
Wassenaar, T.M. (1997) Toxin production by Campylobacter spp. Clin Microbiol Rev. 10: 466–76.
Whyte, P., Mc Gill, K., Cowley, D., Madden, R.H., Moran, L., Scates, P., Carroll, C., OLeary, A., Fanning, S., Collins, J.D., Mc Namara, E., Moore, J.E., Cormican, M.  (2004) Occurrence of Campylobacter in retail foods in Ireland. Inter. J Food Microbiol. 95: 111-118.
Wieczorek, K., Szewczyk, R., Osek, J. (2012) Prevalence, antimicrobial resistance, and molecular characterization of Campylobacter jejuni and C. coli isolated from retail raw meat in Poland. Vet Med. 57: 293–299.
Yildirim, M., Istanbulluoglu, E., Ayvali, B. (2005) Prevalence and antibitoic susceptibility of thermophilic Campylobacter species in broiler chickens. Turk J Vet Anim Sci. 29: 655-660.