Detection of cytolethal distending toxin (cdt) genes of Campylobacter Jejuni and Coli in fecal samples of pet birds in Iran


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


BACKGROUND: Campylobacteriosis is one of the most common
bacterial intestinal disorders of humans in many countries.
A wide range of domestic and wild bird species have been identified
as natural reservoirs. OBJECTIVES: The aim of this study
was to determine the Campylobacter spp from pet birds and
screen the determined Campylobacters for presence of virulence
cytolethal distending toxin (cdt) genes. METHODS: A total
of 660 fecal samples from 32 different species of pet birds were
taken and examined for detection of Campylobacter spp. and
were investigated for presence of cdt genes. All the samples
were collected from clinically healthy birds that were kept in
cage, zoological parks, and/ or in zoo of Tehran, the capital of
Iran. RESULTS: In total, 20 thermophilic Campylobacter were
detected from 8 different avian species. From 20 confirmed
Campylobacter spp., 16 samples (80%) were C. jejuni positive
and 4 (20%) were C. coli in species-specific PCR test. Furthermore,
out of 20 detected Campylobacter, 13 (65%) harbored
the various subunits of cdtA, cdtB and cdtC genes, and 7 (35%)
were negative for all tested cdt genes. CONCLUSIONS: Our findings
indicate that the carriage rate of Campylobacter in different
species of cage and/or in zoo birds is high and confirm that
cdt genes may frequently be present in campylobacter spp.


Article Title [Persian]

شناسایی ژنهای حدت cdt کمپیلوباکترهای ژوژنی و کلی از نمونههای مدفوعی پرندگان خانگی (زینتی) ایران

Authors [Persian]

  • فاطمه احسان نژاد
  • ارژنگ شیخ الملوکی
  • محمد حسن زاده
  • روجا شجاعی کاوان
  • محمد سلطانی
گروه بیماریهای پرندگان، دانشکده دامپزشکی دانشگاه تهران، تهران، ایران
Abstract [Persian]

زمینه مطالعه: کمپیلوباکتر‌ها در بسیاری از موارد سبب آنتریت انسانی می‌شوند. تعداد زیادی از پرندگان اهلی و وحشی مخزن کمپیلوباکترها بوده و دارای ژن‌های حدت cdt می‌باشند. هدف: در این مطالعه ابتدا کمپیلوباکترهای ژژونی و کلی در پرندگان خانگی شناسایی و سپس حضور ژن‌های cdt در آنها مورد مطالعه قرار گرفت. روش کار: تعداد 660 نمونه سواب مدفوعی، از 32 گونه مختلف پرنده خانگی یا زینتی که به کلینیک دانشکده دامپزشکی مراجعه یا در قفس‌های پارک‌ها و باغ وحش تهران نگهداری می‌شدند تهیه گردید. با استفاده از روش باکتریولوژیک و PCR ابتدا کمپیلوباکترها شناسایی و سپس ژن‌های cdt آنها مطالعه گردید. ضمناً محصولات PCR برای سکانس ژن ارسال گردید. نتایج: تعداد 20 نمونه کمپیلوباکتر شامل 16 نمونه ژژونی (80%) و 4 نمونه کلی (20%) از 8 گونه مختلف پرنده خانگی جدا و یا با روش PCR شناسایی و تأیید گردید. در 20 نمونه مثبت، تعداد 13 نمونه (65%) حداقل دارای یک یا چند ژن حدت cdt بودند و 7 نمونه از آنها هیچ یک از ژن‌های cdt را در مطالعه مولکولی نشان ندادند. نتایج اولیه سکانس ژن‌های حدت موید این بود که قرابت بالایی بین ژن‌های حدت این باکتری‌ها با ژن‌های حاصل از جدایه‌های انسانی و طیور صنعتی وجود دارد.  نتیجه‌گیری‌نهایی: مطالعه حاضر نشان داد که پرندگان خانگی آلوده به کمپیلوباکتر بوده و حاوی ژن‌های حدت هستند و می‌توانند بعنوان مخزن باکتری محسوب و سبب آلودگی سایر پرندگان و انسان‌ها بشوند.

Keywords [Persian]

  • کمپیلوباکتر گرمادوست
  • ژن حدت
  • پرندگان زینتی
Abuoun, M., Manning, G., Cawthraw, S., Ridley,
A., Ahmed, I., Wassenaar, T., Newell, D. (2005)
Cytolethal distending toxin (CDT)-negative
Campylobacter jejuni strains and anti-CDT
neutralizing antibodies are induced during human
infection but not during colonization in
chickens. Infect Immun. 72: 3053–3062.
Abulreesh, H.H., Paget, T.A., Goulder, R.
(2006) Campylobacter in waterfowl and aquatic
environments: incidence and methods of
detection. Environ Sci Technol. 40: 7122–7131.
Abulreesh, H.H., Goulder, R. Scott, G.W.
(2007) Wild birds and human pathogens in
the context of ringing and migration. Ringing
and Migration. 33: 193-200.
ALLAN, B. (2006) Effect of cytolethal distending
toxin of Campylobacter jejuni on adhesion
and internalization in cultured cells and in
colonization of the chicken gut. Avian Dis. 50:
Ansari- Lari, M. Hosseinzadeh, S. Shekarforoush.
S.S. Abdollahi, M. Berizi, E. (2011)
Prevalence and risk factors associated with
Campylobacter infections in broiler flocks in
Shiraz. Shiraz, southern Iran. Int J Food Microbiol.
144: 475-479.
Asakura, M., Samosornsuk, W., Taguchi, M.,
Kobayashi, K., Misawa, N., Kusumoto, M.,
Nishimura, K., Matsuhisa, A. Yamasaki, S.
(2007) Comparative analysis of cytolethal
distending toxin (cdt) genes among Campylobacter
jejuni, C. coli, and C. fetus strains. Microb
Pathog. 42: 174–183.
Asakura, M., Samosornsuk, W., Hinenoya, A.,
Misawa, N., Nishimura, K., Matsuhisa, A. Yamasaki,
S. (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. Immunol
Med Microbiol. 52: 260-266.
Avrain, L., Humbert, F., LHospitalier, R., Sanders,
P., Vernozy-Rozand, C., Kempf, I. (2003)
Antimicrobial resistance in Campylobacter
from broilers: association with production
type and antimicrobial use. Vet Microbiol. 96:
Blaser, M.J. (1997) Epidemiologic and clinical
features of Campylobacter jejuni infections.
J Infect Dis. 176: S103–S105.
Carter, P.E., McTavish, S.M., Brooks, D.,
Campbell, J.M., Collins-Emerson, A.C., Midwinter,
M. (2009) Novel clonal complexes with
an unknown animal reservoir dominate Campylobacter
jejuni isolates from river water in
New Zealand. Appl Environ Microbiol. 75:
Bang, 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 Microliol. 50:
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.
Hanninen, M., Pajarre, S., Klessner, M., Rauteline,
H. (1998) Typing of human Campylobacter
jejuni isolates in Finland by pulsedfield
gel electrophoresis. J Clin Microbiol. 36:
Hermans, D., Van Deun, K., Martel, A., Van
Immerseel, F., Messens, W., Heyndrickx, M.,
Haesebrouck,. F. Pasmans., F. (2011) Colonization
factors of Campylobacter jejuni in the
chicken gut. Vet Res. 42:82.
Hickey, T., Majam, G., Guerry, P. (2005) Intracelluar
survival of Campylobacter jejuni
in human monocytic cells and induction of
apoptotic death by cytolethal distending toxin.
Infect Immun. 73: 5194–5197.
Jeffrey, J.S., Atwill, E.R., Hunter, A. (2001)
Prevalence of Campylobacter and Salmonella
at a squab (Young Pigeon) processing plant.
Poul Sci. 80: 151–155.
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. J Clin Microbiol. 37:
Lutful Kabir, S.M., Kikuchi, K., Asakura, M.,
Shiramaru, S., Tsuruoka, N., Goto, A., Hinenoya,
A., Yamasaki, S.H. (2011) Evaluation of
cytolethal distending toxin (cdt) gene-based
species-specific multiplex PCR assay for the
identification of Campylobacter strain isolated
from diarrheal patients in Japan. Jpn J Infect
Dis. 64: 19-27.
Lara-Tejero, M., Galàn, J.E. (2001) cdtA, cdtB,
and cdtC form a tripartite complex that is required
for cytolethal distending toxin activity.
Infect Immun. 69: 4358–4365.
Lara-Tejero, M., Galàn, J.E. (2002) Cytolethal
distending toxin: limited damage as a strategy
to modulate cellular functions. Trends Microbiol.
10: 147–52.
Lee, R., Hassane, D., Cottle, D.L., Pickett, C.
(2003) Interactions of Campylobacter jejuni
cytolethal distending toxin subunits cdtA and
cdtC with HeLa cells. Infect Immun. 71: 4883–
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.
Mullner, P., Jones, G., Noble, A., Spencer,
S.E.F., Hathaway, S., French, N.P. (2009)
Source attribution of food-borne zoonoses in
New Zealand: a modified Hald model. Risk
Anal. 29: 970–984.
Megraud, F. (1997) Isolation of Campylobacter
spp. from Pigeon Feces by a Combined Enrichment-
Filtration Technique. Appl Environ
Microbiol. 53: 1394-1395
Mirzaie, M., 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.
Newell, D.G. (2001) Animal models of Campylobacter
jejuni colonization and disease and
the lessons to be learned from similar Helicobacter
pylori models. J Appl Microbiol. 90:
Newell, D.G., Fearnley, C. (2003) Sources of
Campylobacter colonization in broiler chickens.
Appl Environ Microbiol. 69: 4343–4351.
Newell, D.G., Wagenaar, J.A. (2000) Poultry
infections and their control at the farm level,
In: Campylobacter. Nachamkin, I., Blaser, M.
J. (eds.). (2nd ed.) American Society for Microbiol
Press, Washington, D.C., USA. p. 497–
Nuijten, P.J., van den Berg, A.J., Formentini,
I., van der Zeijst, B.A., Jacobs, A.A. (2000)
DNA rearrangements in the flagellin locus of
an flaA mutant of Campylobacter jejuni during
colonization of chicken ceca. Infect Immunol
68: 7137-7140.
Pasquali, F., DE Cesare, A., Manfreda, G., Frahini,
A. (2011) Campylobacter control strategies
in European poultry production. World’s
Poul Sci J. 67: 1-18.
On, S.L.W., Holmes, B. (1992) Assessment of
enzyme detection tests useful in identification
of campylobacteria. J Clin Microbiol. 30: 746-
Pickett, C.L., Whitehouse, C.A. (1999) The
cytolethal distending toxin family. Trends Microbiol.
7: 292–297.
Rahimi, E., Momtaz, H., Bonyadian, M.
(2010) PCR detection of Campylobacter spp.
from turkey carcasses during processing plant
in Iran. Food Control. 21: 692- 694.
Rizal, A., Kumar, A., Vidyart, A.S. (2010)
Prevalence of pathogenic genes in Campylobacter
jejuni isolated from poultry and human.
Inter J Food Safet. 12: 29-34.
Shane, S.M. (1992) The significance of infection
in poultry: a review. Avian Pathol. 21:
Tamura, K., Peterson, D., Peterson., N., Stecher.,
G., Nei., M., Kumar, S. (2011) MEGA5:
Molecular evolutionary genetics analysis using
maximum likelihood, evolutionary distance,
and maximum parsimony methods. Mol Biol
Evol. 31: 1-9.
Waldenstrom, J., Broman, T., Carlsson, I., Hasselquist,
D., Achterberg, R.P., Wagenaar, J.A.,
Olsen, B. (2002) Prevalence of Campylobacter
jejuni, Campylobacter lari, and Campylobacter
coli in different ecological guilds and taxa of
migrating birds. Appl Environ Microbiol. 68:
Waldenstrom, J., Mevius, D., Veldman, K.,
Broman, T., Hasselquist, D., Olsen, B. (2005)
Antimicrobial resistance profiles of Campylobacter
jejuni isolates from wild birds in Sweden.
Appl Environ Microbiol. 71: 2438–2441.
Yogasundram, K., Shane, S.M., Harrington,
K.S. (1989) Prevalence of Campylobacter jejuni
in selected domestic and wild birds in Louisiana.
Avian Dis. 33: 664–667.