Award details

Development of multiplexed diagnostic biosensor for infectious reproductive diseases of cattle and buffaloes

Reference	BB/L00464X/1
Principal Investigator / Supervisor	Professor Jonathan Cooper
Co-Investigators / Co-Supervisors	Dr Julien Reboud
Institution	University of Glasgow
Department	School of Engineering
Funding type	Research
Value (£)	437,440
Status	Completed
Type	Research Grant
Start date	19/05/2014
End date	18/11/2017
Duration	42 months

Abstract

The proposal brings together Biomedical Engineering in the University of Glasgow (who have recently developed and commercialised new integrated immuno- and DNA based biosensors for low cost diagnostics), OIE/WHO Collaborating Centre, Animal Health and Veterinary Laboratory Agency, Weybridge (who are an OIE reference laboratory for Brucellosis), with the Indian Veterinary Research Institute, Izatnagar (who will develop assays and perform field trials). Researchers at the IVRI will express candidate antigens, specific to individual pathogens, with recombinant technologies using a prokaryotic expression system. B-cell epitopes will be identified and synthetic peptides then synthesized. Additionally, smooth lipopolysaccharide (LPS) and synthetic o-polysaccharide (OPS), of Brucella origin, will be prepared. The immuno-reactivity of the antigens, peptides, LPS and OPS will be assessed and suitable ligands selected to develop a novel multiplexed immunosensor platform. In Glasgow, technology development will take our patented technology for diagnostics, based upon ultrasonics, from pre-prototype instrumentation to a standalone system that performs an assay at the point of care (a farm or AI station). As part of this, three further important themes will be developed: the integration of sensing and sample preparation within the same instrument using a single transducing mechanism, i.e. ultrasonics; transfer of ttechnology to very low cost diagnostic chips (based upon laminated card); and the development of DNA/antibody probe immobilisation technology for the low cost chips. Working together, IVRI and Glasgow will design DNA-specific probe/nucleic acid amplification platforms to directly detect pathogens in milk, semen, uterine and nasal discharges of cattle and buffaloes. Included in this new low cost "point of care" diagnostic platform will be a new patented method for integrating sample preparation from crude samples, enabling access to DNA from cellular materials.

Summary

India is the top dairy producer in the world (yielding about 125 billion litres per annum from cattle and buffaloes). This milk is of huge importance in providing nutrition to a growing population - whilst also providing economic benefit in rural communities. Sustaining this so-called "white revolution" is a vital component in agricultural policy. Similarly in the UK, milk plays an important part in the rural economy and in providing nutrition to the population. In both cases, the supply of cows, to enable the production of this large volume of milk is enabled through the breeding of cattle through both natural and artificial insemination. Maintaining the reproductive health of the cattle and buffaloes is hugely important and diseases that reduce the yield of new calves born have a significant economic impact. The most common reproductive diseases in cattle and buffaloes are brucellosis, leptospirosis and infectious bovine rhinotracheitis. The diseases affect the livestock's reproductive organs, reducing fertility and resulting in a loss of milk production - with a significant impact on agricultural output. These diseases all have a particularly high prevalence in India, and other Asian, African and South American countries as well as being of significant economic importance in the Developed World, including the UK. In addition, leptospirosis and brucellosis are two important bacterial diseases, which have a zoonotic importance - as animal workers, veterinarians, abattoir workers and farmers are also at a high risk of infection. All three diseases are characterized by reproductive failure through abortions, stillbirths and the birth of weak offspring. Infected bulls transmit the pathogens to healthy cows through semen during both natural and artificial insemination. Simple observation by animal owners and producers is not sufficient to accurately diagnose the disease, which often goes unnoticed. We therefore plan to develop low-cost, rapid and sensitive multiplexed diagnostic assays for the three diseases, with the aim of screening cows in the dairy herd and bulls at the artificial insemination station - where such diagnostics will have great impact. Our overall aim is to help break the cycle of infection and control rates of transmission in dairy animals. The major outcome of the project will be a low cost, rapid and sensitive diagnostic test for detection of multiple infectious pathogens affecting reproductive system of cattle and buffaloes. The sensor will be designed and produced in such a way that untrained rural farmers will be able to perform the tests -giving a simple yes or no to tell whether the bull or its semen is infected. The instrument itself will be rugidised so that it can work in the conditions on the farm.

Impact Summary

Our overall aim to create impact within society through improvement in diagnosis and treatment of disease in veterinary science, where this is presently constrained by difficulties associated with the complexity of automation of sample preparation. This situation arises both in India and in the UK This impact will be realised in practice through the development of commercial devices and intellectual property, the delivery of which will have additional economic impacts - with translation of the technologies into medical healthcare challenges. Societal Impact: The most common reproductive diseases in cattle and buffaloes are brucellosis, leptospirosis and infectious bovine rhinotracheitis (IBR). The diseases affect the livestock's reproductive organs, reducing fertility and resulting in a loss of milk production. These diseases all have a particularly high prevalence in India, and other Asian, African and South American countries as well as being of significant economic importance in the Developed World, including the UK. They are particularly important in India which is the top dairy producer in the world (~125 billion litres p.a.). Sustaining this "white revolution" is a vital component in agricultural policy, requiring new diagnostic methods to help break the cycle of infection and control rates of transmission in dairy animals. In addition, leptospirosis and brucellosis are two important bacterial diseases which have a zoonotic importance. In this proposal we plan to develop low-cost, rapid and sensitive multiplexed diagnostic assays for the three diseases (bovine brucellosis, leptospirosis and IBR) with the aim of screening cows in the dairy herd and bulls at the AI station - where such diagnostics will have the greatest impact. The path to uptake is ensured in the project through our links into Indian and UK Veterinary Services and Research Centres, allowing us to test prototypes in relevant settings and demonstrate the ability of the devices to be manufactured in a scalable process. The project allows a number of different pathways to be explored in infectious disease diagnosis. This both reduces risk in the project, by presenting different challenges, and widens opportunity for uptake. Future uptake ultimately relies upon commercialisation of devices. Our partnerships with Veterinary Services and the collection of robust data from animals will steer research towards delivering prototypes that force us to consider manufacturing issues at an early stage in the design process, helping to shorten the route to market. Equally important in this aspect are proposed partnerships with Regional Development Agencies (such as Scottish Enterprise or locally in India), where it is intended that their understanding of industry needs for the new materials will again shorten the route to market. Economic Impact and Beneficiaries: The research, above, is likely to have its most significant impact on livestock (with improved healthcare and outcomes), on veterinary scientists, milk production and farmers (developing new techniques and new methodologies) and societal benefits including the local economy. Using the example of milk sustainability, we will illustrate the potential impact. Beyond veterinary diagnostics, the platform also has potential application in a broad range of diagnostic opportunities, in medical sensing, in either the home, hospital ward or laboratory e.g. for HIV, HCV, meningitis, MRSA, Group A Strep., as well as infectious diseases of the Developing World. This particular opportunity of veterinary diagnostics also affords opportunities for further collaboration, additional EU and TSB funding and for licensing opportunities, all benefiting the economy. Whilst we hope to create a spin-out company in medical diagnostics, there is the opportunity that these activities could in the future be drawn into a single portfolio, providing scale and breadth, or alternatively be realized through a separate activity

Committee	Research Committee A (Animal disease, health and welfare)
Research Topics	Animal Health, Immunology, Microbiology, Technology and Methods Development
Research Priority	X – Research Priority information not available
Research Initiative	Farmed Animal Disease and Health (FADH) [2013]
Funding Scheme	X – not Funded via a specific Funding Scheme

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