A metabonomic approach to identify biomarkers for bovine mycotoxicosis

Lead Research Organisation: Imperial College London
Department Name: Surgery and Cancer

Abstract

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

Technical Summary

Mycotoxicosis is a disease associated with livestock following ingestion of mycotoxins produced through fungal/mould contamination of feed. The disease manifests itself through reduced livestock performance reflecting in altered nutrient metabolism, endocrine malfunctions and immune-suppression. However it is difficult to diagnose at an early stage due to the commonality of symptoms with other production related diseases (e.g. SARA). Mycotoxin production is predominately associated with cereals and grains and hence there are current EU guidelines suggest maximum limits for two mycotoxins - deoxynivalenol and zearalenone - in feed grains and complete feeds. These specifically target reduction of mycotoxin contamination in human food and monogastric livestock feed (pigs and poultry) which are predominately fed on cereal and grains. However this does not cover the wider range of mycotoxins produced from not only fungal/mould contamination of grain and cereal but other feedstuffs e.g. straw and whole crop silage. It also does not investigate the impact of these mycotoxins on ruminants. A recent veterinary survey in the UK showed a high incidence of mycotoxicosis in dairy and beef herds associated with sub-standard aerobically spoiled maize and grass silage. The problem does not end in animal disease or production losses as mycotoxins in the feed of dairy animals can lead to their metabolic products in dairy products, which pose a risk in human health. This project will investigate the effect of mycotoxins associated with ruminant feed and mycotoxin ruminal metabolites on ruminal gastro-intestinal epithelial cells using metabonomic approaches (NMR). This will identify an array of key metabolites associated with mycotoxin poisoning of ruminants which will then be used in vivo to identify biomarkers in bodily fluid (plasma, urine and saliva) which could be used for early diagnosis of the disease.

Planned Impact

Academic Impact -
As highlighted under academic beneficiaries the project will produce a high level of high impact academic findings. We will disseminate our results to the academic community by publications in high impact journals and presentations at conferences and meetings (in the areas of animal nutrition, cell biology, immunology, food research and microbial interactions). Duchy College are part of the project team who are experts in Knowledge transfer and outreach and will organise farm visit days and veterinary workshops where the latest more applied outputs from projects are disseminated to end users.

Economic and societal impacts -
It is envisaged that findings from this project will stimulate development of technology with support from UK and EU funding agencies, which will benefit our industry partners who are part of the Industrial Partnership Award as well as the wider industrial community.
The industry partners in the project include:

a) Micron-Biosystem Ltd. has a dedicated focus on researching techniques for the remediation of mycotoxin contamination in animal feedstuffs and holds patents and product registrations in this field. The company has recently completed the in-house development of methodologies based upon UPLC-MS/MS for the determination of a large suite of mycotoxins in a range of body fluids as well as feedstuffs. These techniques are being intensively utilised in both internal and collaborative R&D projects on three continents. Their involvement within the project will assist in the analysis of mycotoxins and their metabolites.

b) AB Vista Ltd. Is part of Associated British Foods (ABF) and is an international supplier of world leading feed ingredients and technical services. AB Vista's feed micro-ingredients are among the most advanced in the world and are sold across five continents. ABF is one of Europe's largest food companies employing 102,000 people in 46 countries around the world. Links with other companies in the ABF group allow consistent supply of reliable feed ingredients of the highest quality. These products help improve the nutritive value of animal feeds and help reduce the environmental impact while improving production efficiency, and end product quality. ABF's unrivalled expertise in animal feeds and experience of introducing new feed ingredients enables AB Vista to help customers optimise their returns when using its products.

c) Mole Valley Farmers Ltd. are a farmers co-operative which supply as well as feed and products on farm nutritional advice. Their nutritionists have significant experience of practical day to day on farm rationing, including issues such as mycotoxicosis and have regular veterinary contact. The nutritionist role also incorporates co-ordination of nutritional policy, evaluating new products and strategies, and liaising with Mole Valley Forage Services (MVFS). MVFS link forage production and rationing which have excellent geographical coverage.

The three industrial partners in the project will be instrumental in processing the information produced on mycotoxins biomarkers in ruminants into an applied project for industrial up-take. The industrial partners will each produce an exploitation plan which will form a key part of the projects pathway to impact

Further societal impacts from the project will involve improved farm animal health and reduced environmental pollution through increased knowledge at the farm level of the importance of high quality feed production. This will reduce feed waste and also reduce effluent loss from silage as farmers will be encouraged to produce high quality less mycotoxin containing silage. The findings will also ultimately benefit the general public by improving the safety of the food they consume as an early diagnosis of mycotoxicosis in animals will limit the chances of mycotoxins and their metabolites being transferred to animal products and subsequently into the human food chain.

Publications

10 25 50
 
Description The project is still in its early phases but initial results indicate a presence of Patulin mycotoxin in maize silage. Also the project is developing novel cell lines of rumen epithelial cells to test for biomarkers - this is going well currently. Update: We have identified that deoxynivalenol can impact on the ruminal microbiota preventing its ability to produce methanol and potentially increasing the presence of dimethylamine in the gut. As such, dimethylamine may serve as a sub-clinical biomarker for deoxynivalenol. The in vivo trial has finished, all of the metabolic data has been acquired and the data analysis phase is underway.

Update: Clear metabolic variation associated with sub-clinical mycotoxin exposure has been identified in cows. From this data candidate biomarkers for DON exposure have been identified in a number of different sample types. This includes: sarcosine and formate in urine, butyrate in saliva, and 3-phenylpropionate, dimethyl-sulfone, acetate, butyrate, propionate, ethanol and isovalerate in the ruminal contents. The findings of the in vitro and in vivo studies are being prepared for publication as are the findings from the ruminal microbiota study.
Exploitation Route Plans are underway, in collaboration with Mole Valley, for a farm wide survey to test the efficacy of the urine biomarker (sarcosine) identified in the in vivo study. Validation of this promising non-invasive biomarker has great potential for an economic impact by providing a diagnostic test to identify mycotoxin exposed cows earlier to prevent loss of productivity.
Sectors Agriculture

Food and Drink

URL http://www.bovmycotox.co.uk/about-the-project/progress/
 
Description The findings from this study were presented at a consultation meeting for veterinarians at Bicton College in June 2017. This lecture/workshop informed the vets of the cutting edge metabolic profiling technologies currently being used in the field of veterinary science and agriculture. It also provided an update on the metabolic impact of mycotoxins on the gut microbial and host metabolism of cows. At the BovMycoTox conference we disseminated our research findings to the industry experts. This included the protective effects of a binder from Micron-Biosystems on gut microbiota following mycotoxin exposure. We also presented our findings on candidate biomarkers for sub-clinical mycotoxicosis in cows.
First Year Of Impact 2017
Sector Agriculture, Food and Drink
Impact Types Economic

Policy & public services

 
Description Contribution to Parliamentary Office of Science and Technology (POST) note - The Microbiome and Human Health
Geographic Reach National 
Policy Influence Type Implementation circular/rapid advice/letter to e.g. Ministry of Health
URL https://post.parliament.uk/research-briefings/post-pn-0574/
 
Description Wrote an article for GPs detailing the importance of considering the gut microbiota in drug therapy
Geographic Reach National 
Policy Influence Type Implementation circular/rapid advice/letter to e.g. Ministry of Health
URL https://www.gponline.com/viewpoint-understanding-gut-microbiome-impact-drugs/article/1787621
 
Description Understanding how food and beverages deliver improved nutrition across the lifecourse
Amount £1,900,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 11/2022 
End 10/2025
 
Title Bovine saliva metabolic profiling 
Description Developed an 1H NMR spectroscopy method to measure the metabolic profile of bovine saliva samples and ruminal contents. This involves both sample preparation and NMR experiment optimisation. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? No  
Impact Using this approach candidate biomarkers for bovine mycotoxicosis have been identified. In saliva, butyrate was decreased with DON exposure and in the ruminal contents DON increased the abundance of 3-phenylpropionate, uracil, dimethyl-sulfone, butyrate, propionate, and ethanol. 
 
Description AB Vista 
Organisation AB Vista
Country United Kingdom 
Sector Private 
PI Contribution Attendance and input at project meetings: 5/5/2015 1st meeting 4/11/2015 2nd meeting 17/5/2016 3rd meeting 23/11/2016 4th meeting Expertise and facilities to carry out metabonomic analysis of samples generated under Milestone 1.5. Analysis of data generated during this process.
Collaborator Contribution Attendance and input at project meetings: 5/5/2015 1st meeting 4/11/2015 2nd meeting 17/5/2016 3rd meeting 23/11/2016 4th meeting
Impact None as yet
Start Year 2016
 
Description Bristol University 
Organisation University of Bristol
Department School of Social and Community Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Attendance and input at project meetings: 5/5/2015 1st meeting 4/11/2015 2nd meeting 17/5/2016 3rd meeting 23/11/2016 4th meeting. Expertise and facilities to perform metabonomic analysis of samples generated under Milestone 1.5. Analysis of data generated during this process.
Collaborator Contribution Expertise in cell culture techniques and access to facilities for this. Bovine epithelial cells were cultured in vitro in the presence or absence of individual mycotoxins (deoxynivalenol, zearalenone, fumonisin B1 and patulin). Cell culture supernatants and lysed cells were harvested to determine the metabolic profile of cells under normal conditions or treated with mycotoxins as proposed in Milestone 1.5.
Impact None as yet
Start Year 2016
 
Description Micron Bio-Systems 
Organisation Micron Bio-Systems
Country United Kingdom 
Sector Private 
PI Contribution Attendance and input at project meetings: 5/5/2015 1st meeting 4/11/2015 2nd meeting 17/5/2016 3rd meeting 23/11/2016 4th meeting. Expertise and facilities to perform metabonomic analysis of samples generated under Milestone 1.5. Analysis of data generated during this process.
Collaborator Contribution April 2015 - January 2016 Expertise in the field of mycotoxin analysis and analysis of samples for M1. Development of production of mycotoxins in vitro in such a way as to ensure they are closely representative to that produced in the field. Development of mycotoxin solutions appropriate for applying to cells for M1.4 and animals M4: purchase of mycotoxins, expertise in diluents and solvents appropriate for analysis.
Impact The expertise of Micron Bio-Systems in mycotoxin measurements has been sought to measure samples from a separate commercially funded project exploring the role of the diet in the development of Cushings disease in horses. This is a project funded by EquiLife in collaboration with the Veterinary School at the University of Surrey.
Start Year 2016
 
Description Mole Valley Feed Company 
Organisation Mole Valley Farmers
Country United Kingdom 
Sector Private 
PI Contribution Attendance and input at project meetings: 5/5/2015 1st meeting 4/11/2015 2nd meeting 17/5/2016 3rd meeting 23/11/2016 4th meeting. Expertise and facilities to perform metabonomic analysis of samples generated under Milestone 1.5. Analysis of data generated during this process.
Collaborator Contribution Advice on feeds given to dairy animals at different stages of production and what will be appropriate for the in vivo work Supply of maize for in vitro production of mycotoxins (M1) and concentrate for batch culture (M2) Attendance and input at project meetings: 5/5/2015 1st meeting 4/11/2015 2nd meeting 17/5/2016 3rd meeting 23/11/2016 4th meeting
Impact None as yet
Start Year 2016
 
Description Prof Clara Belzer at Wageningen University 
Organisation Wageningen University & Research
Country Netherlands 
Sector Academic/University 
PI Contribution Following some exciting research findings about the potential role of gut microbial metabolites in the early-life development of autism spectrum disorders, a collaboration has been set up (mediated by Danone Nutricia) with Prof Clara Belzer at Wageningen. A PhD student (Patrick Schimmel) working in Prof Belzer's research team is spending 9 weeks (commencing March 2022) at Southampton performing bacterial metabolomics to confirm the findings from our human study.
Collaborator Contribution The group at Wageningen has performed several culture experiments (mono- and co-cultures) and collected culture supernatant for metabolomics.
Impact Placement of a PhD student. PhD student has trained other lab members in bacterial culture techniques.
Start Year 2021
 
Description Delivered a Metabolomics workshop to the Federal University of Ceara, Brazil 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact A workshop on metabolomics was delivered to researchers and clinicians based at the Federal University of Ceara in Northeastern Brazil. This two day workshop covered analytical chemistry, multivariate statistics, and biological interpretation of results. All statistical tools used were open access and the research groups had access to necessary analytical chemistry platforms so this added value to their research.
Year(s) Of Engagement Activity 2020
 
Description Farmer Discussion Group 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Industry/Business
Results and Impact Michael Lee (Project PI) led an open farm discussion event on issues relating to mycotoxins and silage quality.
Year(s) Of Engagement Activity 2015
 
Description Invited talk and chairperson at the 14th European Nutrition Conference FENS in Belgrade, Serbia 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Delivered a 20 minute talk at the 14th European Nutrition Conference FENS in Belgrade, Serbia titled "Measuring bacterial metabolites in biological samples - what can this tell us about the gut microbiome?". I was also invited to act as the chairperson for a session on fatty acids and health.
Year(s) Of Engagement Activity 2023
URL https://fens2023.org
 
Description Invited talk for the European Society for Clinical Nutrition and Metabolism annual meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited talk at the 45th ESPEN Congress on Clinical Nutrition and Metabolism in Lyon, France 11-14 September 2023. I delivered a 30 minute talk on 'Metabolomic and metagenomic signatures in chronic disease'.
Year(s) Of Engagement Activity 2023
URL https://espen.org/files/2023-45th_ESPEN_1st_Announcement.pdf
 
Description Launch of Project website 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Launch of the projects website to disseminate information and inform of the issues facing farmers with mycotoxicosis
Year(s) Of Engagement Activity 2016
URL http://www.bovmycotox.co.uk/
 
Description Nestle Education workshop on the Microbiota 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Undergraduate students
Results and Impact Designed and recorded teaching material for an online course providing information on the gut microbiota, malnutrition and its role in the gut-brain axis. This was in collaboration with Nestle and was aimed and a varied audience mainly based in LMIC countries.
Year(s) Of Engagement Activity 2022,2023
 
Description Newsletter to Farmers 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact A newsletter was circulated in both the Mole Valley magazine and Rural Business School of Duchy College newsletter which is distributed to over 10,000 practitioners highlighting the project
Year(s) Of Engagement Activity 2016
URL https://www.ruralbusinessschool.org.uk/news/breaking-the-mould
 
Description Plenary lecture at the Nordic Metabolomics Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited to give the plenary talk at the Nordic Metabolomics Society annual meeting in Copenhagen. It was attended by a broad audience related to metabolomics research and this talk focused on the metabolic interactions between the microbiome and the mammalian host.
Year(s) Of Engagement Activity 2022
 
Description Scientific Advisory Board for the RENEW project on the Gut-Bone Axis led by Prof Hanne Bertram (Aarhus University) 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact A member of the Scientific Advisory Board for the RENEW project on the role of calcium absorption on the Gut-Bone axis funded by Innovation Fund Denmark. Providing expertise on the application of metabolomics and the gut microbiota.
Year(s) Of Engagement Activity 2020,2021
URL https://pure.au.dk/portal/en/projects/renew--valorisering-af-sidestroemme-fra-mejeriindustrien-til-f...
 
Description VET CONSULTATION MEETING AT BICTON COLLEGE 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Lecture on study findings and an introduction to metabolic phenotyping in veterinary sciences
Year(s) Of Engagement Activity 2017
URL http://www.bovmycotox.co.uk/about-the-project/progress/
 
Description Virtual talk for 'Science' journal online 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Provided an online talk on the gut microbiota for the Science webinar series. This is popular webinar series with a broad global reach. I was contacted by individuals after the talk about potential collaborations.
Year(s) Of Engagement Activity 2022
URL https://www.science.org