Developing liquid AP-MALDI MS as a rapid large-scale classification method for determining farm animal health

Good farm animal health is paramount for food safety and security. It also reduces the burden on the environment and public health by a reduction in disease treatments using antimicrobials or other drugs. Greater knowledge and earlier detection of agriculturally important diseases will result in better farm disease management and welfare of farm animals as well as improved food produce yield and safety. In addition, recent food adulteration scandals have highlighted the need for faster and more detailed knowledge of our food and its production line to determine and guarantee food authenticity.

Information gathering for all of the above is still limited by the analytical methods available. Some of these are not sufficiently cost-effective and/or too cumbersome and slow for rapid action. Others can be fast but are often very targeted, allowing the detection of only one condition, and limited in their practical use, particularly in combination with other tests.

Fortunately, advances in modern mass spectrometry now allow highly sensitive recordings of the molecular profiles of biological samples such as milk. In combination with advanced bioinformatics these recordings can be used to teach computer algorithms for future classification and thus detection of agriculturally important conditions such as mastitis. As modern mass spectrometry is one of the most sensitive molecular detection and characterisation techniques, allowing the analysis of many molecular biomarkers simultaneously, there is now a great opportunity to make a step change in the early detection of such markers for many diseases in a cost-effective and fast way.
However, cost-effectiveness and speed of analysis can still be limited due to the sample preparation necessary for analysis by mass spectrometry. Our lab has recently demonstrated that biological samples such as milk can be effectively analysed by mass spectrometry with no or minimal sample preparation by using liquid MALDI mass spectrometry.

In collaboration with the University of Reading's Department of Agriculture and Centre of Dairy Research (CEDAR) with its large research herd of around 600 cows, we now aim to explore and fully develop a rapid analysis workflow for biological fluids from farm animals based on modern liquid MALDI mass spectrometry. This workflow should allow for early and specific detection of agriculturally important diseases and conditions, exploiting to the fullest mass spectrometry's great potential in highly sensitive, fast and inexpensive characterisation of the health of our farm animals and their produce.

Good farm animal health is paramount for food safety and security. It also reduces the burden on the environment and public health by a reduction in disease treatments using antimicrobial and other drugs. Greater knowledge and earlier detection of agriculturally important diseases will result in better farm disease management and welfare of farm animals as well as improved food produce yield and safety.
Information gathering for the above is still limited by the analytical methods available, often due to cost or a lack of robustness, accuracy or speed. Some methods such as ELISAs can be fast but are targeted, allowing the detection of only one condition.

We have recently developed liquid AP-MALDI MS as a new rapid analysis method using the well-established MALDI technique but allowing for the analysis of crude liquids such as milk with no or minimal ('one-pot') sample preparation. We have also used this new method in atmospheric pressure (AP) ionisation sources of three commercial instruments with great success, thus circumventing the requirement for time-consuming sample introduction to a high-vacuum ion source. Both changes have enabled us to substantially reduce the time needed per sample to be analysed, now allowing highly sensitive, accurate and rapid recordings of the molecular profiles of biofluids such as milk. In combination with advanced bioinformatics these recordings can be used to teach computer algorithms for the detection and classification of agriculturally important conditions.

In collaboration with the University of Reading's Department of Agriculture and Centre of Dairy Research (CEDAR) with its large research herd of ~600 cows, we will develop and validate this new methodology. The developed workflow should allow for early and specific detection of agriculturally important diseases and conditions, exploiting to the fullest mass spectrometry's great potential in highly sensitive, fast and inexpensive characterisation of farm animal health.

Farm animal and plant health is widely discussed in the public and arguably an area where great improvements can still be made. Through accurate (and if possible earlier) diagnosis of diseases on our farms substantial improvements will be achievable in many areas such as the reduction of antimicrobial treatments and pest control agents. Advances in these are a crucial component in combating antimicrobial resistance and environmental pollution. They also provide improved animal health and welfare and an environment that is greener, more robust and diverse. These are desirable outcomes that naturally attract interest and benefit the public. In addition, farming can be more cost-effective if diagnostic testing is faster, cheaper (but more accurate) and multiplexed.

Consequently, the impact of the proposed work of advancing farm diagnostics to the next level of analytical capabilities using novel and more powerful methods can be extremely rewarding.

Scientists and farmers will benefit from the direct and indirect scientific and agricultural impacts of the proposed research, which include:
- More accurate and potentially earlier (pre-symptomatic) analysis of farm animal and crop diseases.
- Less testing (through the potential of multiplexing) but more diagnostic information
- Reduction in the application of disease treatments, including antimicrobial treatments
- Less environmental pollution
- Discovery of new research targets and leads for improved animal and crop health and their identification and characterization
- Further input for the development of a comprehensive model for pathways in agriculturally important diseases

However, the public (and farmers and the food industry) will also benefit through the direct and indirect societal and economic impacts of the proposal, which include:
- Cost-reduction in farming by reducing treatments and pest control and ultimately lower costs of diagnostic testing
- Greener farming with all its benefits to the environment
- Improved consumer confidence and public opinion towards farming
- Population health benefits through a reduction in the risk of antimicrobial resistance using 'precision medicine' for farm animals (lowering the number of unnecessary treatments)