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Animal botulism: innovative tools for diagnosis, prevention, control and epidemiological investigation.

Topic: Disease control and surveillance
Duration of project: 36 Months
Total project costs: 1.717.603€
Animal group: Cattle, Poultry, Others

Project partners

  1. Agence nationale de sécurité sanitaire de l’alimentation, de l’environnement et du travail (ANSES), France
  2. Istituto Zooprofilattico Sperimentale delle Venezie, Italy
  3. Istituto Zooprofilattico Sperimentale delle Regioni Lazio e Toscana, Italy
  4. Statens veterinärmedicinska anstalt (SVA)/National Veterinary Institute, Sweden
  5. Friedrich-Loeffler-Institut, Germany
  6. Central Veterinary Institute of Wageningen UR, Netherlands
  7. Robert Koch-Institut, Germany
  8. Faculty of Veterinary Medicine, Finland

Abstract

Animal botulism is a re-emerging problem worldwide that concerns several species (cattle, mink, horses and birds) and both livestock production and wildlife. This leads to huge economical losses in the animal industry because of high mortality rates. It also presents a risk for transmission to other species, including humans. Despite being reported for a long time, many aspects of the disease have been neglected up to now, in particular approaches for diagnosis and surveillance of botulism have to be improved and harmonized and control and prevention measures have to be developed. This project aims first at developing an alternative approach to the mouse bioassay, which today is still the gold standard for botulism diagnosis because of the lack of a validated in vitro assay. An animal replacement method based on mass spectrometry (Endopep-MS) will be improved and standardized to lead to a sensitive and rapid test for laboratory diagnosis.

This project will also explore the epidemiological aspects of animal botulism focusing on potential risk factors associated with the outbreaks for better managing animal botulism surveillance systems. As a useful tool intended for molecular epidemiology and for the assessment of genetic diversity of C. botulinum group III organisms, Multiple Locus of Variable tandem repeat Analysis and Multilocus Sequence Typing protocols will be developed and whole genome sequencing will be performed. In addition, the sequence variability of botulinum neurotoxins will be determined using mass spectrometry. Usefulness of this approach for epidemiological applications will be evaluated.

Finally, we will focus on the development of prevention and control strategies by testing three strategies: vaccination, use of lactic acid bacteria as antagonist of C. botulinum group III organism growth and toxinogenesis and set up consolidated guidelines for sampling and laboratory testing in botulism outbreaks.

This collaborative 36-month project involves 8 research groups from EU with complementary expertise in C. botulinum, botulinum neurotoxins, mass spectrometry, veterinary diagnostics, genomic studies, epidemiology, and animal experiments. This project will allow a prompt diagnosis of animal botulism, will make available molecular tools which are essential to react early in case of major outbreaks, will clarify essential epidemiological aspects of botulism in poultry and bovine production in Europe, and finally will propose countermeasures.

The main objectives of the project are:

  1. To develop, validate, and standardize the chemical/biochemical method Endopep-MS for confirmation of the botulism diagnosis in animal or environmental samples. The goal is for the Endopep-MS method to replace the mouse bioassay, which is still considered to be the gold standard for detection of botulinum neurotoxins.
  2. To study the epidemiological aspects of botulism in animals and develop monitoring systems. Potential risk factors associated with outbreaks should be identified in order to achieve better management of botulism outbreaks in the future.
  3. To improve the prevention of botulism outbreaks by testing different strategies: vaccination, use of lactic acid bacteria to prevent the growth of botulinum neurotoxin producing bacteria, and to develop common guidelines for sampling and laboratory tests in botulism outbreaks.

The main goals of the project are:

  1. To enable a rapid diagnosis of botulism in animals.
  2. To make available molecular tools that are necessary to respond early to major botulism outbreaks
  3. To clarify important epidemiological aspects of botulism in production animals in Europe.
  4. To propose preventive measures to avoid / limit botulism outbreaks.

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