Advanced Metagenomics, Sensors and Photocatalysis for Antimicrobial Resistance Elimination (AMSPARE)

Lead Research Organisation: University of the West of Scotland
Department Name: School of Health & Life Sciences

Abstract

This proposal brings together experts on sensor technologies, water treatment and remediation from India with experts on environmental microbiology, meta-omics geochemistry and policy and industrial regulatory processes from the UK, to engage the issue of AMR proliferation in the environment. Specifically, we will focus on the potential for increased AMR due to aggravation by pharmaceutical waste entering waterways. This research would engage with the Indian pharmaceutical industry to study the AMR production 'metabolism', within a complex interplay of environmental geochemical and microbiological processes, in order to refine policy and improve regulatory control in pharmaceutical waste management.

In brief, the inter-disciplinary research team from the UK and India will assess the life cycle of pharmaceutical waste water through analysis of chemical and AMR profiling surrounding small to medium pharmaceutical plants that discharge waste into Common Effluent Treatment Plants (CETPs). CETPS are the main focus in this study for waste water antimicrobial contamination. Large plants now are committed to solid waste disposal. Microbial profiling will include abundance of antibiotic resistant genes and effects of complex microbial communities through a combination of metagenomics, digital droplet PCR and culture studies.

In parallel, waste 'clean up' will be addressed through the the employment of low cost sensors to detect what residues are present in the waste and affordable photocatalytic technology to remove the residues before the effluent is discharged into the environment. Validation of these technologies will be carried out under laboratory conditions using site samples and maintaining the chemical and microbial profile obtained from our mapping work.

Finally, due to the sensitive issues surrounding this project engagement with regulatory bodies and industry is critical. Utilising our data and technology we will be able to influence the creation of policies that work for all stakeholders, including local communities and the environment, thus reducing the impact of antimicrobial waste in the environment.

Planned Impact

We seek to utilize low-cost sensor and photocatalytic technologies to 'clean up' the waste effluent and to determine what outcomes this has on the environmental health. Furthermore, we aim to sustain engagement with beneficiaries of this research based on the 'One Health' approach principles. Therefore, impact is integrated into the project proposed. Impacts will be seen in Policy development and regulation, Pharmaceutical Industry, Government of India and other LMICs, Public Understanding as well as the Scientific Community.
Policy/regulatory impact - Impact will be measured by the co-production of outcome planning and integration of our results into regulatory changes and 'buy-in' from Small and Medium Enterprises that utilise Common Effluent Treatment Plants (CETPs) to discharge waste.

Pharmaceutical Industry and beyond- Engagement with Industry and Government is essential for the progress of this project and impact will be assessed by the engagement of new technologies and open discussions of how to improve the process. In addition, the reach and transfer of this technology will have impacts in other areas for example in urban sewage systems, clinical waste management for Hospitals and also the food industry (decontamination of antibiotics from meats and dairy). Impact will be assessed on the development of 'Knowledge Transfer Partnership'-type relationships with other industries and in other LMICs.


Public understanding - Engagement with the Public (Schools, Local Communities, Families, Social clubs) is important to ensure that all community members understand how they can contribute in decreasing the risk on the emergence of AMR in the environment. A range of dissemination materials associated with AMR and use of antibiotics, waste and environmental treatment of emerging pollutants, resource efficiency as a tool for environmental improvement will be created. This will involve the tools described below - including online downloads, outreach packs STEM engagement events in local Schools and communities. Impact will be assessed by developing a questionnaire post event for participants to gauge comprehension and levels of awareness.

Scientific community - We aim to involve and integrate undergraduate and postgraduate teaching programmes, which align directly to the study. There will be extensive research exchange and training, such as student summer placements and researcher visits and development of interdisciplinary skills. The interdisciplinary nature of the project also is a focus for capacity building and establishing future collaborations, reporting to conferences and interest groups (e.g. Student vacation scholarships, research pools in Scotland, NERC clusters, professional societies). Impact will be assessed by analysing the number of student and researcher exchanges and integration events initiated in the three year period.
 
Description the project is not yet completed, but already we have established:
1. antibiotic residues are present in the environment
2. free-living amoebae are habouring antimicrobial resistant Pseudomonas bacteria (and others). This is relevant to human health
3. we are use a point of care device to to identify antibiotics in environmental samples
4. We have established a stakeholder network platform
Exploitation Route The point of care device could be use by industry, public health authorities etc. to monitor antibiotic pollution levels. Researchers can use the knowledge of the importance of microbial ecosystems (and not just bacteria) in further investigations.
Sectors Communities and Social Services/Policy,Environment,Healthcare,Government, Democracy and Justice,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology

URL https://indiaukamrenvironment.org/
 
Description we have used our findings to engage industry, local governments and authorities and NGOs in workshops (January 2023 in Scotland and India) to understand their views on AMR and how they contribute.
First Year Of Impact 2023
Sector Environment,Government, Democracy and Justice,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology
Impact Types Cultural,Societal,Economic,Policy & public services

 
Description Assessment of the role of bacterial endosymbionts on the survival and pathogenicity of the brain eating amoeba, Naegleria fowleri
Amount £5,200 (GBP)
Organisation Scottish Universities Life Sciences Alliance 
Sector Academic/University
Country United Kingdom
Start 06/2022 
End 09/2022
 
Description Developing techniques to study Neoparamoeba species isolated from sea urchins (Ref SERS1-17)
Amount £5,300 (GBP)
Organisation Marine Alliance for Science and Technology for Scotland 
Sector Academic/University
Country United Kingdom
Start 06/2022 
End 12/2022
 
Description Impact of organic pollutants on the emergence of antimicrobial resistance in aquatic environments
Amount £10,000 (GBP)
Organisation Scottish Alliance for Geoscience, Environment and Society (SAGES) 
Sector Academic/University
Country United Kingdom
Start 04/2022 
End 01/2023
 
Title Isolation of Acanthamoeba from contaminated sediments 
Description Isolation of Acanthamoeba sp. from sediments on agar plates 1) Prepare the solution of 2g of sediment and 4mL of deionised water. Mix it very well in tube. 2) filter the solution on filter paper (11uL waterman no1). Put filter paper onto nonnutrient amoeba saline agar plate. Incubate it for 1-2 weeks on 25o C. 3) Check plate for growth. Cut a small piece of agar with amoeba and streak it around new nonnutrient amoeba saline agar plate to get a clean culture of amoeba. Use nonnutrient amoeba saline agar plates with Ecoazole to avoid Fungi contamination if is necessary. Repeat step 3 until you don't get a clean culture. Isolation of Acanthamoeba sp. from sediments in wells 1) Day 1: Cut out a small piece of agar with Amoebae sp. Inoculate it into encystment media with 0,5% PenStrep. Check media for Amoeba sp. growth. 2) Day 2: After 24h wash well with PBS. Take all of the encystment media out (store it in falcon tube for bacterial analysis), add 1mL of PBS, incubate it for 3-5 min, then remove PBS and add 1-2 mL of encystment media with 1% of PenStrep. Add 50uL of PG into well. 3) Next day check wells for Amoeba sp. Growth and possible contamination. Exchange Encystment media with 1% PenStrep, if is necessary. Add 100 uL of PG. 4) Day 4: Check wells for contamination and trophozoite growth. 5) Day 5: Wash wells with Encystment media with 1% of PenStrep. Add 50uL of PG. Incubate wells for 2 days (weekend) in incubator. 6) Day 8: Check wells, exchange media if is necessary; add 100uL of PG. 7) Check wells every day. Continue with washing them with Encystment media+1% PenStrep and adding PG; until they multiply to final CFU/mL. If you have Fungi contamination, use 2% of PenStrep and remove piece of agar out from the well. 
Type Of Material Biological samples 
Year Produced 2017 
Provided To Others? No  
Impact we now have extra data and tools to further study AMR in the environment as Acanthamoeba habour bacteria 
 
Title core sampling for ICP and DNA analysis 
Description the challenge was to extract a sediment core without contaminating it with further DNA and keeping its structure intact for ICP and DNA analysis. 
Type Of Material Technology assay or reagent 
Provided To Others? No  
Impact This technique will help other researchers to perform similar core sample in similar environmental areas 
 
Description 1st Workshop - AMSPARE (Advanced Metagenomics, Sensors and Photocatalysis for Antimicrobial Resistance) 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact update of project including activities, results and future direction. An expert panel including industry engaged in a round table discussion
Year(s) Of Engagement Activity 2021
 
Description AMR methods workshop Programme Coordination Team 
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 workshop to develop consensus for methods in AMR research
Year(s) Of Engagement Activity 2021
 
Description Royal Society of Edinburgh funded Scotland-India network related to tackling antimicrobial residues within the environment 
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 SIAMREN is a network of researchers within Scotland and India who have common research interests in the challenges presented by the emergence of antimicrobial resistance (AMR) in the environment. The network currently consists of researchers in Scotland at: The University of Strathclyde (UoS), The University of the West of Scotland (UWS), The James Hutton Institute, Glasgow Caledonian University, Edinburgh Napier University and Heriot-Watt University, who have projects to study the emergence of AMR in the environment. Heriot-Watt, UWS and UoS, groups already have established partnerships with colleagues from different groups with Indian Institutions. We plan to establish the SIAMREN network through the two events outlined below. SIAMREN is currently funded through the Royal Society of Edinburgh Scottish Asia Partnerships Higher Education Research Fund.

Scope and Theme Workshop (Mid/Late Feb 2021)
This will be focused upon the core grant collaborators and non-academic partners. It is designed to identify key themes and common Scotland/India-related challenges around Antimicrobial Resistance (AMR) in the environment. Each partner will be asked to present one slide on the key problems they face in terms of environmental AMR, or the support they believe they could offer. This will be followed by a discussion of the challenges in a global context in order to identify common research themes for both Scotland and India. The initial workshop will be limited to Scottish collaborators to maintain focus and keep it manageable.

Mini-conference (May 2021)
In May we will run a virtual mini-conference as an interface between environmental AMR activities in Scotland and India. My suggestion is to run this as a morning session - maybe with selected non-academic presentations from stakeholders in India and Scotland, and then an afternoon session with presentations from academics (including early careers researchers) with current projects related to AMR in the environment. The conference will be free for anyone to register so we can raise the profile of the network and also some of the research activities that are taking place in Scotland and in India.
Year(s) Of Engagement Activity 2021