Blanco, J. L.

Diaz, C. Rodriguez

Drigo, I.

Indra, A.

Janezic, S.

Jiraskova, A.

Kokotovic, Branko

Ocepek, M.

Pardon, B.

Perreten, V.

Poxton, I. R.

Rupnik, M.

Seyboldt, C.

Songer, J. G.

Weese, J. S.

Zidaric, V.

Online Research Database In Technology

                       General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.  • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal   If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.     Downloaded from orbit.dtu.dk on: Dec 20, 2017International Clostridium difficile Animal Strain CollectionZidaric, V.; Janezic, S.; Pardon, B.; Indra, A.; Kokotovic, Branko; Blanco, J. L.; Seyboldt, C.; Diaz, C.Rodriguez ; Poxton, I. R.; Perreten, V.; Drigo, I.; Jiraskova, A.; Ocepek, M.; Weese, J. S.; Songer, J. G.;Rupnik, M.Publication date:2012Document VersionPublisher's PDF, also known as Version of recordLink back to DTU OrbitCitation (APA):Zidaric, V., Janezic, S., Pardon, B., Indra, A., Kokotovic, B., Blanco, J. L., ... Rupnik, M. (2012). InternationalClostridium difficile Animal Strain Collection. Poster session presented at 4th International Clostridium difficileSymposium (ICDS 2012), Bled, Slovenia. All 100 included strains were distributed into 39 different PCR ribotypes. Up to 17 different PCR ribotypes can be found within a single animal species and up to 15 different PCR ribotypes per country.   Within one standard (agarose based) PCR ribotype several PCR ribotypes could be distinguished by capillary gel based PCR ribotyping (Table 1; PCR ribotypes 014/020, 078, 002, 045, 056, 033).  Five strains are nontoxigenic while toxigenic strains account for 95.0% and belong to 10 different toxinotypes: 0, I, III, IV, V, VI, VIII, XI, XII and XIX. PCR ribotypes 078, 126, 014/020, 012 and 002 that are frequently associated with animals ( Keel et al., 2007; Janezic et al., 2012) represent 40.0 % of all strains.  V. Zidaric, S. Janezic1, B. Pardon2, A. Indra3, B. Kokotovic4, J.L. Blanco5, C. Seyboldt6, C. Rodriguez Diaz7, I.R, Poxton8, V. Perreten9, I. Drigo10, A. Jiraskova11, M. Ocepek12, J.S. Weese13, J.G. Songer14, M. Rupnik1, 15,16 C. difficile  strains:  Altogether 100 strains from 12 different countries were contributed. Collected strains originate from 11 different animal species, including pets, horses and food animals. Approximately half (58,0%) of the strains are from cattle and pigs (Table 1).   For  every strain aditional availible information on animal host and strain was obtained.   Animals have been recognized as an important potential reservoir of Clostridium difficile and according to recent studies the overlap between PCR ribotypes of human and animal isolates seems to be increasing (Bakker et al., 2010; Gould and Limbago, 2010; Janezic et al., 2012; Keel et al., 2007; Koene et al., 2011). Here we report on an International Clostridium difficile animal strain collection that was established to enhance comparative studies on animal-associated strains and contribute to interlaboratory exchange of the strains.  The goal of the collection is to include one PCR ribotype per species per country/laboratory.  Country                 Animal host Austria Belgium Canada Czech Republic Denmark  Germany Italy Scotland Slovenia Spain Switzerland U.S.A. Total number of strains per animal species Number of different  PCR ribotypes per animal species  Pig XII/056 [new RT] 0/011/049 [049/1]  0/150 [AI-12]     V/078 [078] 0/002 [002/0] 0/081 [081]  V/078 [078] 0/015 [211]  0/150 [AI-12]  V/078 [new RT] V/045 [598] V/126 [126] VI/066 [413] 0/150 [AI-12] 0/005 [005] VI/SLO 137 [new RT]   V/078 [251] 0/150 [AI-12] XIX/012 [012] 0/014/020 [014 subtype] 0/081 [081] 0/SLO 036 [050/AI-84] XII/SLO 133 [AI-15] VI/SLO 012 [new RT] V/078 [078]   V/045 [045] V/078 [078]   V/078 [078] 0/002 [nd] 29  17 Cattle V/078 [078] 0/014/020 [014/0] 0/029 [029] 0/005 [005] 0/015 [211] 0/SLO 125 [014 subtype] 0/SLO 036 [AI-84/050] 0/SLO 090 [AI-9-1] 0/SLO 143 [610] 0/SLO 164 [nd] 0/SLO 165 [nd] V/078 [078]  V/078 [078] V/126 [078 ecdc] XI/033 [033] 0/012 [012] 0/014/020 [014/0] 0/002 [002/2] 0/081 [081] 0/SLO 074 [new RT] V/078 [078]       V/126 [126] XIb/033 [033]     0/002 [209] 0/014/020 [014/0]   XIb/283 [new RT]   V/078 [078] XIa/033 [033] III/027 [027] 29  15 Dog       0/001/072 [nd] 0/002 [002/0]     0/SLO 066 [AI-60] 0/014/020 [014/0] V/SLO 024 [new RT] Tox-/010 [010]       0/014/020 [new RT] 0/012 [012] XII/056 [new RT] Tox-/010 [010]     10 8 Cat       0/001/072 [001 ecdc]     V/SLO 024 [new RT] 0/014/020 [449] Tox-/SLO 002 [AI-34]     0/014/020 [new RT]       5 4 Poultry 0/001/072 [001]                  0/029 [029] 0/014/020 [014/0] IV/023 [023] 0/103 [AI-82/1] 0/001/072 [001]       6  5 Horse     0/014/020 [020] XIb/033 [new RT] V/078 [078] III/027 [027]            XI/033 [nd]     V/078 [078] V/126 [126]   7 5 Rabbit/Hare               0/012 [012] V/078 [078] 0/014/020 [014/0] Tox-/SLO 132 [new RT] 0/SLO 090 [AI-9-1] 0/002 [002/0] VIII/017 [017] Tox-/SLO 084 [205]           8  8 Partridge                   XII/056 [446] XI/033 [new RT]         2 2 Goose                   0/SLO 090 [AI-9-1]       1 1 Crow                   0/003 [003]       1 1 Raccoon     I/SLO 166 [nd] 0/103 [AI-82/1]                    2 2 Total number of strains per country  15  12  12  1  7  7  18  1  16  4  2  5   100 Number of different  PCR ribotypes per country  15 7 9 1 7 5 15 1 12 4 2 4 INTERNATIONAL Clostridium difficile ANIMAL STRAIN COLLECTION Results and Discussion Materials and methods  References  1Institute for Public Health Maribor, Maribor, Slovenia; 2Ghent University, Faculty of Veterinary Medicine, Merelbeke, Belgium; 3AGES, Vienna, Austria; 4Technical University of Denmark, National Veterinary Institute, Copenhagen, Denmark; 5Complutense University, Madrid, Spain; 6Friedrich-Loeffler Institute, Jena ,Germany; 7University of Liege, Faculty of Veterinary Medicine, Liege, Belgium; 8University of Edinburgh, Edinburgh, UK; 9University of Bern, Institute of Veterinary Bacteriology, Bern, Switzerland; 10IZSVe, Treviso, Italy; 11Charles University in Prague, 1st Faculty of Medicine, Prague, Czech Republic; 12University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia; 13University of Guelph, Ontario Veterinary College, Ontario, Canada; 14Iowa State University, Ames, U.S.A.; 15University of Maribor, Medical Faculty, Maribor, Slovenia; 16Centre of excellence for integrated approaches in chemistry and biology of proteins, Ljubljana, Slovenia   Background   Molecular characterization of C. difficile strains:  All collected strains were characterized by toxinotyping (Rupnik et al., 1998; http://www.mf.uni-mb.si/tox/). In adition, binary toxin genes were detected by PCR as described in Stubbs et al. (2000). Standard agarose gel-based PCR ribotyping was used as described by Bidet et al. (1999) and results analyzed by BioNumerics software 5.10 (Applied Maths). Strains were also typed by capillary gel electrophoresis-based ribotyping using primers for standard agarose gel-based PCR ribotyping with flourescein labelled 16S primer (Indra et al., 2008). PCR ribotype patterns were analyzed and identified on a web-based database Webribo (http://webribo.ages.at).   Table 1. Distribution of C. difficile genotypes by species and country given as toxinotype/standard PCR ribotype [capillary gel electrophoresis based PCR ribotype].   Bakker D, Corver J, Harmanus C, Goorhuis A, Keessen EC, Fawley WN, Wilcox MH, Kuijper EJ. 2010. Relatedness of human and animal Clostridium difficile PCR ribotype 078 isolates determined on the basis of Multilocus Variable-Number Tandem-Repeat Analysis and tetracycline resistance. J. Clin. Microbiol. 48(10):3744–3749. Bidet  P,  Barbut  F,  Lalande  V,  Burghoffer  B,  Petit  JC. 1999. Development  of  a  new  PCR-ribotyping  method  for  Clostridium  difficile  based  on  ribosomal  RNA  gene  sequencing. FEMS  Microbiol. Lett. 175:261-266.  Gould LH, Limbago B. 2010. Clostridium difficile in food and domestic animals: a new foodborne pathogen? Clin Infect Dis. 51(5):577-582. Indra A, Huhulescu S, Schneeweis M, Hasenberger P, Kernbichler S, Fiedler A, Wewalka G, Allerberger F, Kuijper EJ. 2008. Characterization of Clostridium difficile isolates using capillary gel electrophoresis-based PCR ribotyping. J. Med. Microbiol. 57: 1377-1382.      Janezic S, Ocepek M, Zidaric V, Rupnik M. 2012. Clostridium difficile genotypes other than ribotype 078 that are prevalent among human, animal and environmental isolates. BMC Microbiol. 12:48 doi:10.1186/1471-2180-12-48       Keel K, Brazier JS, Post KW, Weese S, Songer JG. 2007. Prevalence of PCR ribotypes among Clostridium difficile isolates from pigs, calves, and other species. J.Clin. Microbiol. 45(6):1963-1964.       Koene MG, Mevius D, Wagenaar JA, Harmanus C, Hensgens MP, Meetsma AM, Putirulan FF, van Bergen MA, Kuijper EJ. 2011. Clostridium difficile in Dutch animals: their presence, characteristics and similarities with human isolates. Clin. Microbiol. Infect. doi:10.1111/j.1469-0691.2011.03651.x       Rupnik M, Avesani V, Janc M, Von Eichel-Streiber C, Delmée M. 1998. A novel toxinotyping scheme and correlation of toxinotypes with serogroups of Clostridium difficile isolates. J. Clin. Microbiol. 36(8):2240-2247.       Stubbs  S,  Rupnik  M,  Gibert  M,  Brazier  J,  Duerden  B,  Popoff  M. 2000. Production  of  actin-specific  ADP-ribosyltransferase  (binary  toxin)  by  strains  of Clostridium  difficile.  FEMS  Microbiol. Lett. 186:307-312.        nd- typing not done ; new RT- PCR ribotype has on webribo database not yet been determined; subtype- subtype has on webribo not yet  been determined   

International <em>Clostridium difficile</em> Animal Strain Collection

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International <em>Clostridium difficile</em> Animal Strain Collection

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