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Clostridium botulinum and C. difficile Research, Botulism Reference Service for Canada

Research Scientist: Richard Harris, PhD
Bureau of Microbial Hazards, Health Canada
richard.harris@hc-sc.gc.ca

Research Activity

Epidemiology of foodborne and infant botulism. Growth and ecology of clostridia in foods.

General Overview

Clostridia are associated with a multitude of human diseases including foodborne and infant botulism, antibiotic-associated diarrhoea and pseudomembranous colitis.

Botulism continues to evolve as a disease. The first case of infant botulism was diagnosed in Canada in 1979. Now, infant botulism has become the most common form of botulism in Canada. The first case of adult intestinal colonization botulism in Canada was recognised in 2006. Since this first case, several additional cases have been recognised.

The importance of the human pathogen C. difficile has been brought to the fore recently as a result of outbreaks of C. difficile associated disease (CDAD). CDAD involves colonization of the intestine, suggesting an involvement of foods in the transmission of the disease. The link with foods has been confirmed by recent reports indicating a high prevalence of C. difficile in Canadian ground meats. There is a requirement for information regarding incidence, control, and elimination of this emerging pathogen in foods.

This project addresses factors determining the risk of foodborne illness due to clostridia. These include: incidence and levels of contamination of foods with clostridia, destruction of C. botulinum and C. difficile in foods, inhibition of growth of C. botulinum by storage temperature, pH, water activity, shelf-life and packaging atmosphere of foods, replacement of the mouse bioassay for botulinum neurotoxin with in vitro alternative assays.

Current Research Activities

  • Incidence and levels of contamination of foods with clostridia
  • Destruction of C. botulinum and C. difficile in foods
  • Inhibition of growth of C. botulinum by storage temperature, pH, water activity and in packaging atmosphere of foods

Laboratory Services

R. Harris, PhD [office (613) 957-0885, mobile: (613) 296-1139]; R. Boone [laboratory (613) 946-6826]; Health Products and Food Branch, Health Canada, Ottawa, Ontario, KIA 0L2, Postal Locator 2204A2.

Recent Publications

  • Harris RA, Flint A, Blondin Brosseau M, Weedmark K, Austin JW. Complete genomes of Clostridium botulinum type B(F) isolates associated with a 1995 foodborne botulism outbreak from commercial pâté reveals a recombination event disrupting the ntnh gene. Microb Genom. 2024 Jan;10(1):001169. doi: 10.1099/mgen.0.001169. PMID: 38175697; PMCID: PMC10868621.
  • Harris RA, Blondin-Brosseau M, Levesque C, Rasmussen PE, Beauchemin S, Austin JW. Viable Clostridium botulinum spores not detected in the household dust of major Canadian cities. Epidemiol Infect. 2023 Sep 7;151:e154. doi: 10.1017/S0950268823001474. PMID: 37675600; PMCID: PMC10548537.
  • Harris RA, Tchao C, Prystajecky N, Weedmark K, Tcholakov Y, Lefebvre M, Austin JW. Foodborne Botulism, Canada, 2006-20211. Emerg Infect Dis. 2023 Sep;29(9):1730–7. doi: 10.3201/eid2909.230409. PMID: 37610295; PMCID: PMC10461667.
  • Assal N, Boone R, Harris RA, Gabriel M, Sasges M, Petri B, Ramaswamy H, Austin JW. Inactivation of Group I and Group II Clostridium botulinum spores by ultraviolet irradiation in water. Int J Food Microbiol. 2023 Jun 16;395:110191. doi: 10.1016/j.ijfoodmicro.2023.110191. Epub 2023 Mar 31. PMID: 37019040.
  • Anniballi F, Smith TJ, Austin JW. Editorial: New insights on botulism, botulinum neurotoxins, and botulinum toxin-producing clostridia. Front Microbiol. 2022 Jul 18;13:979653. doi: 10.3389/fmicb.2022.979653. PMID: 35928165; PMCID: PMC9344910.
  • Cadieux B, Lawal OU, Emond-Rheault JG, Jeukens J, Freschi L, Kukavica-Ibrulj I, Levesque RC, Austin JW, Goodridge L. Draft Genome Sequences of Two Clostridium botulinum Group II Strains Carrying Phage-Like Plasmids. Microbiol Resour Announc. 2022 Jun 16;11(6):e0009122. doi: 10.1128/mra.00091-22. Epub 2022 May 18. PMID: 35583330; PMCID: PMC9202375.
  • Harris R, Tchao C, Prystajecky N, Cutler J, Austin JW. A summary of surveillance, morbidity and microbiology of laboratory-confirmed cases of infant botulism in Canada, 1979-2019. Can Commun Dis Rep. 2021 Jun 8;47(78):322-328. doi: 10.14745/ccdr.v47i78a05. PMID: 34421389; PMCID: PMC8340675.
  • Harris R, Dussault F, Flint A, Austin JW, Weedmark K. Complete Genome Sequence of Clostridium botulinum CJ0611A1, a Type A(B) Isolate Associated with an International Outbreak of Botulism from Commercial Carrot Juice. Microbiol Resour Announc. 2021 Jan 7;10(1):e01111-20. doi: 10.1128/MRA.01111-20. PMID: 33414295; PMCID: PMC8407696.
  • Harris RA, Anniballi F, Austin JW. Adult Intestinal Toxemia Botulism. Toxins (Basel). 2020 Jan 24;12(2):81. doi: 10.3390/toxins12020081. PMID: 31991691; PMCID: PMC7076759.
  • Siu K, Rehan M, Austin JW, Ramachandran Nair R, Pernica J. It's not all about the honey. Paediatr Child Health. 2017 Mar;22(1):37-38. doi: 10.1093/pch/pxx009. Epub 2017 Mar 30. PMID: 29479172; PMCID: PMC5804883.
  • Leclair D, Farber JM, Pagotto F, Suppa S, Doidge B, Austin JW. Tracking sources of Clostridium botulinum type E contamination in seal meat. Int J Circumpolar Health. 2017;76(1):1380994. doi: 10.1080/22423982.2017.1380994. PMID: 28982302; PMCID: PMC5645764.
  • Peck MW, Smith TJ, Anniballi F, Austin JW, Bano L, Bradshaw M, Cuervo P, Cheng LW, Derman Y, Dorner BG, Fisher A, Hill KK, Kalb SR, Korkeala H, Lindström M, Lista F, Lúquez C, Mazuet C, Pirazzini M, Popoff MR, Rossetto O, Rummel A, Sesardic D, Singh BR, Stringer SC. Historical Perspectives and Guidelines for Botulinum Neurotoxin Subtype Nomenclature. Toxins (Basel). 2017 Jan 18;9(1):38. doi: 10.3390/toxins9010038. PMID: 28106761; PMCID: PMC5308270.
  • Carter AT, Austin JW, Weedmark KA, Peck MW. Evolution of Chromosomal Clostridium botulinum Type E Neurotoxin Gene Clusters: Evidence Provided by Their Rare Plasmid-Borne Counterparts. Genome Biol Evol. 2016 Mar 2;8(3):540-55. doi: 10.1093/gbe/evw017. PMID: 26936890; PMCID: PMC4824171.
  • Weedmark KA, Mabon P, Hayden KL, Lambert D, Van Domselaar G, Austin JW, Corbett CR. Clostridium botulinum Group II Isolate Phylogenomic Profiling Using Whole-Genome Sequence Data. Appl Environ Microbiol. 2015 Sep 1;81(17):5938-48. doi: 10.1128/AEM.01155-15. Epub 2015 Jun 26. PMID: 26116673; PMCID: PMC4551264.
  • Walton RN, Clemens A, Chung J, Moore S, Wharton D, Haydu L, de Villa E, Sanders G, Bussey J, Richardson D, Austin JW. Outbreak of type E foodborne botulism linked to traditionally prepared salted fish in Ontario, Canada. Foodborne Pathog Dis. 2014 Oct;11(10):830-4. doi: 10.1089/fpd.2014.1783. Epub 2014 Sep 4. PMID: 25188279.
  • Weedmark KA, Lambert DL, Mabon P, Hayden KL, Urfano CJ, Leclair D, Van Domselaar G, Austin JW, Corbett CR. Two novel toxin variants revealed by whole-genome sequencing of 175 Clostridium botulinum type E strains. Appl Environ Microbiol. 2014 Oct;80(20):6334-45. doi: 10.1128/AEM.01573-14. Epub 2014 Aug 8. PMID: 25107978; PMCID: PMC4178653.
  • Leclair D, Fung J, Isaac-Renton JL, Proulx JF, May-Hadford J, Ellis A, Ashton E, Bekal S, Farber JM, Blanchfield B, Austin JW. Foodborne botulism in Canada, 1985-2005. Emerg Infect Dis. 2013 Jun;19(6):961-8. doi: 10.3201/eid1906.120873. PMID: 23735780; PMCID: PMC3713816.
  • Leclair D, Farber JM, Doidge B, Blanchfield B, Suppa S, Pagotto F, Austin JW. Distribution of Clostridium botulinum type E strains in Nunavik, Northern Quebec, Canada. Appl Environ Microbiol. 2013 Jan;79(2):646-54. doi: 10.1128/AEM.05999-11. Epub 2012 Nov 16. PMID: 23160120; PMCID: PMC3553760.
  • Schwartz KL, Austin JW, Science M. Constipation and poor feeding in an infant with botulism. CMAJ. 2012 Nov 20;184(17):1919-22. doi: 10.1503/cmaj.120340. Epub 2012 Sep 17. PMID: 22988147; PMCID: PMC3503904.
  • Sheppard YD, Middleton D, Whitfield Y, Tyndel F, Haider S, Spiegelman J, Swartz RH, Nelder MP, Baker SL, Landry L, Maceachern R, Deamond S, Ross L, Peters G, Baird M, Rose D, Sanders G, Austin JW. Intestinal toxemia botulism in 3 adults, Ontario, Canada, 2006-2008. Emerg Infect Dis. 2012 Jan;18(1):1-6. doi: 10.3201/eid1801.110533. PMID: 22257757; PMCID: PMC3310098.
  • John W. Austin and Daniel Leclair (2011) Botulism in the North: A Disease Without Borders. Clinical Infectious Diseases 52: 593-594
  • Susan M. Twine, Christopher W. Reid, Annie Aubry, David R. McMullin, Kelly M. Fulton, John Austin & Susan M. Logan (2009) Motility and flagellar glycosylation in Clostridium difficile. Journal of Bacteriology 191(22):7050-62
  • A.T. Carter, C.J. Paul, D.R. Mason, S. M. Twine, M. Alston, S.M. Logan, J.W. Austin, and M. W. Peck (2009) Independent evolution of neurotoxin and flagellar genetic loci in proteolytic Clostridium botulinum. BMC Genomics 10:115
  • Martin L. Kalmokoff, John W. Austin, Terry D. Cyr, Mary A. Hefford, Ronald M. Teather, Leonard Brent Selinger (2009) Physical and genetic characterization of an outer membrane protein (OmpM1) containing an N-terminal S-layer-like homology domain from the phylogenetically Gram-positive gut anaerobe Mitsuokella multacida. Anaerobe 15(3): 74-81
  • Anandi N. Sheth, Petra Wiersma, David Atrubin, Vinita Dubey, Don Zink, Guy Skinner, Fran Doerr, Patricia Juliao, German Gonzalez, Cindy Burnett, Cherie Drenzek, Carrie Shuler, John Austin, Andrea Ellis, Susan Maslanka, Jeremy Sobel. (2008) International outbreak of botulism with prolonged toxemia due to commercial carrot juice. Clinical Infectious Diseases 47(10):1245-51
  • Jamie Spiegelman, David Cescon, Yael Friedman, Brie V. Mazza, John W. Austin, Anita Rachlis and Brian J. Murray (2008) Bowel loops and eyelid droops. Canadian Medical Association Journal 179(9):927-9
  • Susan M. Twine, Catherine J. Paul, Evgeny Vinogradov, David J. McNally, J.R. Brisson, James A. Mullen, David R. McMullin, H. C. Jarrell, John W. Austin, John F. Kelly and Susan M. Logan. (2008) Structural and genetic characterisation of flagellar glycosylation in Clostridium botulinum. FEBS Journal 275: 4428-4444
  • Lu J., P. Delaquis, C. Vigneault, M.T. Charles, G.S.V. Raghavan, V Toussaint and J.W. Austin. (2008) Multidisciplinary approach to postharvest heat treatment of fruits and vegetables. In : C. Stevens and V.A. Khan (Eds). Recent Advances in Agriculture. Research Signpost, Kerala, India. Chapter 8, pp 189-210.
  • Flaman, A. S., H. Albaghdadi, J. McClintock, T. Cutts, A. Hashem, M. LeBrun, J. Austin, E. Brown, R. He, and X. Li (2008) Additive inhibition of H1N1 and H3N2 influenza viruses by aurintricarboxylic acid with amantadine. In Options for the Control of Influenza Virus. Page 478-479. J. Katz (Eds). 2008, ISBN: 978-901-769-15-6. International Medical Press. London. U.K
  • Catherine J. Paul, Shulin Tran, Kevin J. Tam, and John W. Austin. (2007) A unique restriction site in the flaA gene allows rapid differentiation of Group I and Group II Clostridium botulinum strains by PCR and RFLP analysis. J. Food Prot. 70: 2133-2139
  • Delaquis, P. and J. W. Austin (2007) The effect of heat treatments on the fate of foodborne pathogens in horticultural produce. Stewart Postharvest Review. Volume 3, Number 3, June 2007 , pp. 1-5
  • Catherine J. Paul, Susan M. Twine, Kevin J. Tam, James A. Mullen, John F. Kelly, John W. Austin, and Susan M. Logan. (2007) Flagellin diversity in Clostridium botulinum groups I and II: a new strategy for strain identification. Appl. Environ. Microbiol. 73: 2963-75
  • Rhonda KuoLee, Xigeng Zhao, John Austin, Greg Harris, J. Wayne Conlan and Wangxue Chen. (2007) Mouse Model of Oral Infection with Virulent Type A Francisella tularensis. Infection and Immunity 75: 1651-1660
  • Jonah Kirkwood, Andrew Ghetler, Jacqueline Sedman, Daniel Leclair, Franco Pagotto, John Austin and Ashraf Ismail (2006) Differentiation of Clostridium botulinum Strains by Focal Plane Array Fourier Transform Infrared Spectroscopy. J. Food Protection 69: 2377-2383
  • D. Leclair, F. Pagotto, J. M. Farber, B. Cadieux and J. W. Austin (2006) Comparison of DNA fingerprinting methods for use in investigation of type E botulism outbreaks in the Canadian Arctic. J. Clin. Microbiol. 44: 1635-1644
  • Martin Kalmokoff, Patricia Lanthier, Tammy-Lynn Tremblay, Mary Foss, Peter C. Lau, Greg Sanders, John Austin, John Kelly and Christine M. Szymanski. (2006) Proteomic analysis of Campylobacter jejuni 11168 biofilms reveals a role for the motility complex in biofilm formation. J. Bacteriol. 188: 4312-4320
  • Adam M. Yule, Ian K. Barker, John W. Austin and Richard D. Moccia (2006) Toxicity of Clostridium botulinum type E neurotoxin to Great Lakes fishes: Implications for avian botulism. J. Wildlife Dis. 42: 479-493
  • Adam M. Yule, Veronique LePage, John W. Austin, Ian K. Barker and Richard D. Moccia (2006) Repeated low level exposure of the Round Goby (Neogobius melanostomas) to Clostridium botulinum type E neurotoxin. J. Wildlife Dis. 42: 494-500
  • Yule, A.M., Austin J.W., Barker I.K., Cadieux B., and Moccia R.D. (2006) Persistence of Clostridium botulinum neurotoxin type E in tissues from selected freshwater fish species: implications to public health. J. Food Protection. 69:1164-7.
  • Wangxue Chen, Rhonda Kuolee, John Austin, Hua Shen, Yanming Che, and J. Wayne Conlan (2005) Low dose aerosol infection of mice with virulent type A Francisella tularensis induces severe thymus atrophy and CD4+CD8+ thymocyte depletion. Microbial Pathogenesis 39:189-96
  • M. Gauthier, B. Cadieux, J.W. Austin and B.W. Blais (2005) Cloth-Based Hybridization Array System for the Detection of Clostridium botulinum Types A, B, E, and F Neurotoxin Genes. J. Food Prot. 68: 1477-83
  • M.A. Hefford, S. D.Aoust, T.D. Cyr, J.W. Austin, G.Sanders, E.Kheradpir and M.L. Kalmokoff (2005) Proteomic and microscopic analysis of biofilms formed by Listeria monocytogenes 568. Canadian Journal of Microbiology. 51: 197-208 (Micrograph from paper was used for cover of CJM issue)
  • Brigitte Cadieux, Burke Blanchfield, James P. Smith, and John W. Austin (2005) A rapid chemiluminescent slot blot immunoassay for the detection and quantification of Clostridium botulinum neurotoxin type E Int. J. Food Microbiology 101: 9-16

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