Leptospirosis is the fourth most common zoonotic (transmitted from animals to man) disease worldwide and has a number of major farmed, companion and feral animal reservoirs. Vaccination has been widely used to control disease in animals and consequently incidence in humans. Each vaccine batch must be tested for potency before market release. Hamsters are particularly sensitive to leptospirosis and consequently used as a model species to test the potency of vaccine batches. Typically five vaccinated and five unvaccinated hamsters are challenged with virulent (lethal) Leptospira bacteria. Four of the vaccinates must survive and four unvaccinates must show signs of leptospirosis or die. The effects of the disease on hamsters are severe (usually death), the test is time consuming (minimum duration 40 days) and expensive. The RSPCA, pharmaceutical industry and government recognise the pressing need to replace this hamster vaccine batch potency test with a humane alternative. The current proposal seeks to replace the hamster model with alternative generic in vitro (test tube) tests. These will exploit recent technological developments in mass spectrometry enabling the analysis of large biological molecules found in vaccines. The protective components from virulent and avirulent Leptospira bacteria and a commercial vaccine preparation will be separated and detected with antibodies from vaccine immunised hamsters. Active components will be purified, identified and measured by mass spectrometry. This will enable the active components to be compared between batches of vaccines to ensure consistency. A similar approach has been attempted using antibodies to measure vaccine components but these are not widely available and the molecules they measure are poorly defined. In contrast, mass spectrometry is now widely applied across the biological sciences, such methods are transferable and analytes can be readily identified. The Veterinary Laboratories Agency has unique expertise in Leptospira vaccine batch testing and key contacts with industry. This expertise will provide a unique environment for the current project which will focus on the two component canine vaccine, the twin batch test for which is considered to be particularly severe. Dissemination of information will be a critical success factor for the uptake and implementation of any replacement test by vaccine manufactures. This will be achieved by publication in peer reviewed journals, presentation at conferences and discussion with industry contacts. Finally, this new approach to vaccine batch testing could provide an alternative to some of the 35,000 animals currently used for this purpose each year.

Disease and contaminants both pose major risks to wildlife and Man. This is well recognised and there are a variety of surveillance schemes in the UK that monitor wildlife for occurrence and severity of diseases and/or contaminants. These schemes complement rather than duplicate each other but share many operational procedures and so can face similar challenges. The information gathered from each surveillance scheme is communicated to a wide spectrum of end users. The various surveillance schemes are run by different government agencies and laboratories, research centres, institutes and Universities. The funders of the schemes are an equally diverse range of government departments, agencies and industry. A key difficulty caused by this myriad of researchers and funding organisations is that it hampers communication between schemes. The schemes only have opportunistic and ad hoc mechanisms to exchange knowledge or develop common best practices that would facilitate sharing of samples and data. Such cooperation can also be hampered by differences between funders in the priorities that they wish surveillance schemes to address. Furthermore, because each scheme reports its findings largely in isolation, it is difficult for end users to obtain an overview of common or widespread threats. The main aim of this project is to establish a Wildlife Disease & Contaminant Monitoring & Surveillance (WILDCOMS) network. This will provide a partnership between nine current UK contaminant and disease surveillance schemes. The network will foster and facilitate knowledge exchange, harmonisation towards best practice, collaboration and sharing of resources. It will also enhance and widen communication with and between end-users, and in particular will provide end-users with an holistic overview of environmental disease and contaminant risk. This should make identification of emerging hazards and risks easier and quicker to spot, and provide the more integrated scientific evidence base needed to formulate better and timely policy and regulation. The specific objectives, delivered in four work packages, will be: (i) to establish and develop the network through regular partners meetings (ii) to use the network to maximise communication of integrated surveillance information to a wide range of end-users through an annual Stakeholder Forum and through collation of findings from all schemes into web-based quarterly bulletins (iii) development towards harmonised operational procedures (sample collection, measurement, data recording and sample archiving) that will facilitate sharing and collaboration between schemes and eliminate duplication of effort (iv) to develop a sustainable model for WILDCOMS and extend its scope to a European scale through linkage with key European partners and networks WILDCOMS will thus facilitate sharing of skills, expertise, knowledge, samples and data, thereby maximising the use of available resources. This will result in better value for money overall and foster development of new initiatives. The benefits the network will deliver can be summarised as: (a) ntegrated surveillance leading to an improved scientific evidence base with which regulators and policy makers can assess threats to wild vertebrates and human health (b) better long term management, sharing and dissemination of samples, best practice and data (c) a recognised forum that will facilitate discussion and collaboration between surveillance schemes and different end-users and stakeholders (d) an enhanced UK research base by increasing knowledge through scientific publications and greater awareness of activities and specimen archives (e) benefits for industrial end users including potential for averting costs by preventing problems (f) benefits to quality of life to the through improved risk assessment

SARS-CoV-2 infection causes asymptomatic through severe clinical presentations in humans. Parameters around infection and transmission dynamics have been studied in animal models. However, there are growing reports of 'reactivation' and re-infection with variant SARS-CoV-2 viruses, months after 'recovery'. The frequency, timing, clinical and virological consequence, in terms of infectious virus shedding and onward transmission following secondary intra- and inter-species infections, remain unknown. The underlying immunological mechanisms behind these infections also remain unexplored. Additionally, SARS-CoV-2 infection in mink has generated novel virus variants, capable of infecting and partially evading existing immunity in humans, increasing the threats of re-infection. Here, we propose to use our established ferret model of SARS-CoV-2 (human and animal adapted viruses), to study longitudinal outcomes of respiratory droplet infection for two genetically different viruses over a six-month period. We will monitor virological and immunological progression of longitudinal SARS-CoV-2 infection; investigate the consequences of SARS-CoV-2 re-infection with homologous and heterologous strains; and investigate the effect of adaptation and immunity on the evolution of SARS-CoV-2 during these infections. We hypothesise that productive re-infection with SARS-CoV-2 variants can result in both infected and infectious stages, the latter being able to transmit disease irrespective of prior exposure and immune status. These investigations will provide detailed robust novel information relevant to mitigation strategies such as droplet avoidance, herd immunity, vaccination and virus evolution. It will also address questions regarding the emergence and maintenance of SARS-CoV-2 in animal reservoirs which may threaten eradication and long-term management of COVID

Bovine tuberculosis (BTB) presents a significant challenge to animal and public health globally. BTB is also a significant cause of zoonotic TB (zTB), mainly in developing/low and middle income countries (LMIC) as highlighted in the recently published roadmap to zTB eradication jointly produced by WHO, WOAH and the International Union against TB & Lung Disease (UTLD). Their One Health approach acknowledged that TB eradication in animals will impact zTB in humans. Eradication could be accelerated by improved diagnostic tests that overcome limitations associated with Purified Protein Derivative (PPD) tuberculin: limited performance, standardisation, vaccine-interference. These could be overcome by defined antigens. PPDs are used both in tuberculin skin tests and blood interferon-gamma release assays (IGRA). More sensitive blood test platforms that complement the IGRA test could further enhance diagnostic performance. BTB tests primarily target cellular immunity, yet serological tests can also be useful for additional case detection. Thus, our proposal aims to address these constraints to optimise test performance in an inter-connected manner. We will conduct field validation for test sensitivity and specificity of the defined antigen formulation DST-F (fusion protein of ESAT6, CFP10, Rv3615c) in domestic ruminants (cattle, goats, buffalos) in different epidemiological settings (WP1, skin test, IGRA). Environmental exposure to mycobacteria in the farms will be also evaluated. We will develop a multi-cytokine platform (MCP) to enhance test performance beyond the current IGRA, (WP2) and also develop novel antigen capture and lateral flow assays (WP3). The performance of these innovative tests will be assessed with samples generated in WP1. WP4 will critically assess data generated in WP1-3 and provide recommendations for the optimal combinations of test platforms to accelerate BTB eradication.