Title of work

New endpoints for thyroid hormone system disruptor testing with fish

Co-author(s)

Thomas Braunbeck, Lisa Goelz, Pauline Pannetier, Henrik Holbech, Sina Volz, Dries Knapen, Lucia Vergauwen

Introduction

Substantial periods of early vertebrate development are regulated by thyroid hormones (THs) and can therefore easily be disrupted by environmental pollutants, which interact with the TH system (thyroid hormone system-disrupting chemicals, THSDCs). Currently established test systems for environmental assessment of chemicals use amphibian metamorphosis as a sensitive apical endpoint for the detection of THSDCs, e.g. in OECD test guidelines (TG) 231 and 241. For any other endocrine modality, test systems with fish are preferably used, which means that in many cases, multiple animal experiments for the detection of potential THSDCs must be performed due to this separation. Consequently, the lack of thyroid-sensitive endpoints in fish tests has been identified as a serious gap in the currently available battery for endocrine disruptor testing with non-target organisms. Accordingly, different EU-funded research projects aim to close this gap and are working to establish thyroid-sensitive endpoints in fish and implement these into existing fish TGs such as TGs 210, 234 and 236.

Methodology

Extensive literature research has been performed to evaluate the impact of THSDCs on fish and to investigate which of their developmental processes could be disrupted by THSDCs. Based on the results, combined with data from amphibian tests, a set of different model THSDCs has been selected to perform different exposure experiments on fish. Experiments were performed according to the established OECD TGs 210, 229, 234 and 236 or combinations thereof. Wild-type zebrafish (Danio rerio) and a thyroid transgenic line were used. Swim bladder inflation and eye development were selected as the most promising apical endpoints and were combined with analyses of TH levels and thyroid follicle morphology.

Results

In our experiments, we observed strong alterations of thyroid follicle morphology, as well as eye morphology and cellular structure, combined with changes in photoreceptor patterning. Also, swim bladder inflation was severely impaired by different THSDCs. Behavioural analyses demonstrate that these changes directly translate into altered swimming behaviour and visual function of the larvae. Under field conditions, this would most likely result in reduced survival of fish exposed to THSDCs. An AOP network describing the sequence of events from molecular impact on the TH system of fish to population-relevant behavioural changes is already established for swim bladder inflation and is currently being developed for eye development.

Discussion

THSDCs provide strong evidence that eye development and swim bladder inflation, combined with TH level measurements and analyses of thyroid follicle morphology (histology or transgenic lines), can and should be implemented as thyroid-specific endpoints into existing TGs for endocrine disruptor testing with fish. All investigated apical endpoints react sensitively to different modes of action of THSDCs and can be seen as meaningful, population-relevant endpoints. Combined with mechanistic endpoints such as TH level measurements, thyroid follicle histopathology or the use of transgenic thyroid fish lines, they could, in many cases, replace the need to run additional amphibian tests. 

Funded by: 
- the European Union’s Horizon 2020 research and innovation programme, under grant agreement No 825753 (ERGO, Breaking down the wall between human health and environmental testing of endocrine disrupters). 
- European Commission contract No 07.0203/2018/794670/ETU/ENV.B.2 (‘Development of a study protocol for regulatory testing to identify endocrine disrupting substances in biotic systems’).

Title of work

Opportunities for evolving towards a transparent and efficient implementation of the Transparency Regulation

Co-author(s)

Delzenne Pascale, Rocío Fernández Cantón, Martyr Griffiths, Barbara Koch, Miguel Vega-Sanchez 

Introduction

Bayer supports transparency throughout the risk assessment of its products. A company-specific process was initiated in 2017, granting public access to safety-relevant studies used by regulatory authorities to authorise Plant Protection Products (PPPs), and later extended to those used for Genetically Modified (GM) crop risk assessment. The objectives of this initiative at Bayer are therefore similar to those of the subsequently established 2019 EU Transparency Regulation: increase the transparency of the EU risk assessment in the food chain and strengthen the reliability, objectivity and independence of the studies commissioned by industry and assessed by the European Food Safety Authority (EFSA) to increase the EU citizens’ trust in EU food systems. The new Regulation impacts eight food domains overseen by EFSA and of interest to Bayer are GM crops, PPPs and Food Supplements. While the overall objective of the Regulation is supported by Bayer, it provides a high degree of complexity and uncertainty for technology developers that may result in significant delays in the risk assessment that can negatively impact access to innovations and new technologies for farmers and consumers.

Methodology

We developed an overview laying out the different new elements brought by the Transparency Regulation and how these were implemented by EFSA, following the published Practical Arrangement documents and accompanying information. Furthermore, we assessed potential opportunities arising from the Regulation as well as the practical challenges posed by its implementation for technology developers due to the complexity of regulatory data generation as well as other factors such as the existing differences in global regulatory data requirements. Lastly, the assessment provides insights into how Bayer is developing internal systems designed to ensure full compliance with all the new elements of the Regulation.

Results

Our assessment demonstrates that, while the general objectives of the Transparency Regulation are in line with those of Bayer’s transparency initiative initiated in 2017, its implementation can create practical challenges for technology developers. Key challenges were identified with the implementation of the requirement of study notifications, for which the technology developer has to notify, in an EFSA database, each study that supports the application prior to the study start date. Each notified study has to be provided in full upon submission of the application. In the event of non-compliance, justification needs to be provided to EFSA. Regulatory data generation is complex and supports submissions to regulatory agencies of several countries, each of whom have different data requirements. Even though Bayer adapted its internal processes to enable study notifications in compliance with the Regulation, the challenges relating to conflicting submission requirements and timing across the globe may lead to unavoidable compliance challenges with the new requirements for reasons not related to transparency. This can significantly impact the commercialisation of new technologies.

Discussion

If a technology developer is not compliant with the new requirements brought by the Transparency Regulation, even due to reasons not related to transparency, this may lead to procedural consequences, and can be extremely impactful considering the already lengthy timelines of the EU regulatory process, resulting in significant delays in the risk assessment that can limit access to innovations and new technologies for farmers and consumers. While the importance of transparency in the risk assessment is acknowledged and supported by Bayer, this should be done in an efficient and pragmatic manner using processes that enable a consistent dialogue between all stakeholders involved. The elements of such processes that enable a transparent approach should apply equally to all stakeholders, consider the reality, specifics and complexity of regulatory data generation, and guarantee that penalties are only applied when the requirements are not followed for reasons related to transparency.

Title of work

Engaging citizens through deliberative mini publics: evaluation of a German consensus conference on genome editing

Co-author(s)

Introduction

In response to a rising European discourse around ‘erosions of democracy’, more and more organisations are taking a ‘deliberative turn’. Especially within contested fields of emerging technologies, deliberative communication approaches, such as consensus conference or other so-called ‘mini-publics’, have become a prominent tool to engage the public in science-based policy-making.  Mobilising concepts of input, throughout and output legitimacy, this study analyses a recent consensus conference on genome editing initiated by the German Federal Institute for Risk Assessment. Drawing upon observations, surveys and semi-structured interviews with participating citizens, scientists, organisers and invited stakeholders, the presentation illustrates difficulties in ensuring inclusive input from across society. From a processual, or ‘throughput’ perspective, it identifies conflicts between the information requirements emerging from deliberative ideals and empirical information sourcing practice. In terms of output, the conference had its greatest impact on the personal learning of participants. Ensuring an impact on wider policy-making emerged as more challenging.

Methodology

The consensus conference, facilitated by an external communication agency, consisted of two preparation weekends, followed by a public conference where a panel of professional experts provided answers to questions posed by the citizens, based on which the citizens drafted a vote and presented it to stakeholders. An external scientific board reviewed all information material as well as the list of professional experts from which participants could select. Online and radio advertisements resulted in 147 citizen registrations. After categorising them according to socio-demographic criteria, 20 participants were randomly selected from each category. The presenter acted as an observer throughout the conference. All citizen participants received a questionnaire on their experience. In addition, 27 semi-structured interviews were conducted with participating citizens, stakeholders, professional experts and organisational staff. The content of all interview transcripts, observational notes and open questionnaire data was analysed in MAXQDA using legitimacy criteria deducted from the literature as a coding framework. Closed questionnaire data was analysed descriptively using SPSS.

Results

Both interviews and surveys indicate  the difficulties of the conference in ensuring inclusive input from across society with interviewees discussing how the event mainly attracted interested and better-educated parts of society. While the survey suggests that participants were able to include their opinion overall, some interviewees supported allegations of unequally distributed abilities to engage during the process. The majority of citizen participants agreed that processes were transparent and information materials helpful and balanced. A number of citizen interviewees complained though that they were left alone in sourcing additional information. From the organisers perspective, ad-hoc requests for additional information collided with deliberative and stakeholder requests for external review, transparency and independence. When it comes to the question of consensus, pluralistic views on the topic seemed to inhibit the possibility to find a consensus, which, for some interviewees, diluted the ground for a clear impact. The conference showed its greatest impact on the personal learning of participants. Ensuring an impact on wider policy-making emerged as more challenging.

Discussion

Resonating with previous studies, this case illustrates difficulties of deliberative mini-publics in ensuring an output beyond individual learning. Organisers should therefore put greater emphasis on making their results heard. In doing so, they will likely have to continue to balance requests for impact on the one hand and independence on the other hand. While the adherence to deliberative ideals, such as the provision of balanced and externally reviewed information, may help to find this balance, it can be a detriment to the need to facilitate increasingly dynamic information seeking processes. New hybrid formats that combine face-to-face deliberation with internet-based interaction may be able to address some of these challenges. More precisely, they could help participants in their wider information sourcing. If combined with novel advertisement and communication strategies, it may also help attract more diverse citizen groups to address issues of inclusiveness. In advocating for such formats, one should not underestimate the challenges online participation raise though including remaining inclusiveness challenges and their potentially even greater struggle in providing unifying outputs.

Title of work

EU-China Risk Assessment Database for E.coli in Beef

Co-author(s)

Stella Papagianeli, Zafeiro Aspridou, Konstantinos Koutsoumanis

Introduction

Escherichia coli is a major concern for food safety as its pathogenic strains are an etiological agent of serious illness. The primary reservoirs are food producing animals and a lot of infections have been linked to beef and beef products. A lot of research efforts are dedicated to the detection and quantification of the hazard along the path from farm to fork. Nevertheless, this information is not translated into easily accessible data in a well-organised manner which are of high relevance for risk assessment. The recent advancements in global trade have transformed the agri-food chain into a complex system. China is one of the largest markets for European Union food export/import although their existing different approaches to food safety are highlighted. Ensuring and enhancing food safety seems to be a high priority, through information exchange. 
The objective of the present study was the collection of data required for assessing the risk of E.coli in beef and various beef products in the EU and China food chain and the construction of a database providing quantitative data along with information such as food type, food chain stage, packing status, etc.

Methodology

A systematic review was conducted on the Scopus electronic database. The review considered articles published between 1991-2021. Papers were deemed eligible for inclusion based on the microorganism (E.coli), the geographical location (countries of the European Economic Area) and the type of animal (cattle/cows) or product (beef) sampled. Prisma statement (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) was employed for reporting the screening process. After sorting out the duplicates, all collected titles and subsequently abstracts were screened according to the inclusion criteria. Finally, articles with unavailable full text were excluded from the review. The same procedure will be followed for the territory of China.

Results

For Europe, 919 papers were retrieved from Scopus and after the entirety of the screening process, 222 were selected for data extraction. A database was developed and formatted according to the sampling site/stage, the type of product, the method of analysis, and the prevalence and concentration. In addition, various supplementary information for each study was also recorded so as to improve the flexibility of the database. Meta-analysis will be performed and pooled data will be included.

Discussion

The compilation of the data and the construction of the database upon completion will constitute a valuable tool for the development of risk assessment models providing comparative data for the EU and China. The database offers ease of access to well-organised and harmonised as well as pooled data and enables the study of various scenarios along the farm to fork path. Beyond the scientific interest, it is expected to support food safety in the EU-China trade.
Acknowledgments: The study was funded by Project DiTect, ‘Digital Technologies as an enabler for a continuous transformation of food safety system’ (European Union’s Horizon 2020 research and innovation programme under grant agreement No 861915).

Title of work

Improving concentration setting in endocrine screening tests with aquatic vertebrates

Co-author(s)

Laurent Lagadic

Introduction

Concentration-setting guidance (OECD ED test guidelines, ECHA-EFSA ED GD and OECD GD 150) pushes applicants to test as high as possible: just below the level of 10 % mortality. While this makes sense in a risk-based ED assessment (US, Japan), it does not align with the hazard-based cut-off for ED in the EU, where the concern is at the lower end of the concentration response curve. The unreasonable combination of the traditional toxicology paradigm (test high and find a threshold) with the no threshold assumption for ED and thus application of a cut-off can lead to a ban of substances that show ED at concentrations deemed unacceptable in the risk assessment anyway. To address this issue, we suggest an improved concentration-setting practice tailored to the EU, where the focus is on the low end of the concentration response curve. By applying this practice, the improved conc. setting addresses the actual concern more effectively and reduces animal use by: 1. not necessitating range-finding tests and 2. reducing the need for potentially unnecessary higher tier testing. It also helps reduce animal suffering by using concentrations that do not cause mortality (which may trigger a repeat of the ED test).

Methodology

We systematically reviewed the concentration-setting guidance across endocrine testing guidelines and guidance documents and highlighted similarities and inconsistencies.
The improved concentration-setting guidance is based on consistency, available data (acute and chronic studies, including the acute-to-chronic ratio, fish bioconcentration data) and the WEBice tool (to extrapolate from tested to non-tested species), expert judgment and the principles of reducing animal testing and suffering (i.e. avoiding unnecessary range-finder tests and mortality). This paper expands on Wheeler et al. (2013) and takes it further by including additional endocrine guidelines and additional refinements. Lastly, alignment with mammalian toxicology is sought, where the MTD is based on a maximum body weight reduction of 10 % and not mortality.

Results

Based on the principles outlined in the Methodology section, a decision scheme is developed that starts from the available data and which will help observers to determine the maximum test concentration (MTC) and appropriate lower concentrations to be used in the endocrine screening assays.

Discussion

Depending on the available data and their consistency, some expert judgment may still be required. However, overall, the scheme proposed here improves the presently available heterogeneous guidance and aligns the ecotoxicology approach for non-target vertebrates with that used in mammalian toxicology.

Wheeler JR, Panter GH, Weltje L, Thorpe KL. 2013. Test concentration setting for fish in vivo endocrine screening assays. Chemosphere 92(9): 1067-1076. doi: 10.1016/j.chemosphere.2013.01.102.

Title of work

Evolutionary Patterns of Rabies Virus Strains from Georgia Exhibit Risks of Their Transmission and Rabies Coinfections among Dogs and Cattle

Co-author(s)

Ekaterine Gabashvili, Saba Kobakhidze, George Lomidze, Jimsheri Loladze, Levani Tsitskishvili, Mamuka Kotetishvili

Introduction

Rabies is a zoonotic and neurotropic infectious disease caused by members of the Lyssavirus genus with rabies virus (RABV) as its prototype. In Georgia, rabies reflects endemic disease patterns, and has been officially documented since 1930. While RABV-infected dogs remain one of the main sources contributing significantly to the transmission of rabies across both humans and agricultural animals, one third of animal rabies cases in Georgia have been found to occur in agricultural animals, predominantly cattle (83 %). The genotypes and genetic relatedness of RABV isolates that infect dogs, cattle and wild animals in Georgia have been very poorly investigated and need to be urgently determined to gain greater and important insight into the risks of rabies transmission in these animals. Here, we provide an initial insight into the RABV genotypes associated predominantly with dogs and cattle, their population-based genetic structure and relationships, and certain trends of their evolutionary divergence in Georgia.

Methodology

A total of 73 RABV strains recovered from dogs and cattle (including one strain from jackals) in Georgia (2015-2016) were included in our phylogenetic analyses. The DNA sequences of a 1350-bp region of the nucleoprotein gene for these strains were subjected to molecular evolutionary analysis. These DNA sequences were obtained from the nucleotide database of the National Center for Biotechnology Information (NCBI). Using the Basic Local Alignment Search Tool (BLAST), the DNA sequences were blasted against the NCBI nucleotide database to obtain the respective DNA sequences of other RABV isolates recovered globally. The molecular evolutionary analyses were performed using MEGA X. The maximum likelihood (ML) algorithm was used to construct phylogenetic trees and to determine ancestral sequence types (STs). FEL (Fixed Effects Likelihood) and FUBAR (Fast, Unconstrained Bayesian AppRoximation for Inferring Selection) were used to determine the dn/ds ratios and trends of episodic positive/diversifying versus episodic negative/purifying selections, respectively. SplitsTree and SimPlot were employed to determine genetic recombination of nucleoprotein genes between the RABV strains.

Results

The DNA sequence analysis of the nucleoprotein-encoding gene loci discriminated 73 RABV strains from Georgia into 41 STs. Certain strains from dogs and cattle were found to share the same STs (ST1, ST7, ST11, ST37). Two RABV strains (MT079902.1 and MT079888.1) recovered from dogs and jackals were deemed to share ST22. The genetically closest neighbours of the local RABV strains appeared to be those recovered in Turkey (with the ancestral strain determined for the closest relatives of the Georgian STs), Azerbaijan, Russia, Iran, Hungary (with the most recent ancestral strain determined for the closest relatives of the Georgian STs), Tajikistan and Iraq. The local RABV ST30 was determined to be the most recent ancestral ST for the entire subset of strains. The FEL analysis produced dn/ds ratios ≤ 0.05; FUBAR found evidence of episodic negative/purifying selection at 147 sites across the targeted gene loci of the local RABV STs.  The SplitsTree analysis generated five parallelograms shared by multiple RABV strains, with the bootstrap values varying from 85 to 100 (fit: 100). SimPlot could identify recombination breakpoints in the RABV recombined gene loci.

Discussion

Whereas rabies is the endemic zoonotic disease involving dogs and cattle, its transmission risk pathways in agricultural animals have been poorly understood in Georgia. The phylogenetic inferences from our analyses suggest that dogs can be one of the main sources of rabies transmission in cattle in Georgia. Besides, while a direction of rabies transmission between dogs and jackals could not be resolved in the analysis, the latter is assumed to be one of the possible sources of rabies transmission in dog populations in Georgia. Turkey Azerbaijan, Russia, Iran, Hungary, Tajikistan, Iraq and Georgia might be cross-border transmission hotspots for rabies, at least among these regions. The results of our recombination analyses are strongly suggestive of rabies co-infection risks in dog and cattle populations. A small sample size of the RABV strains and the fact that the DNA sequence data are limited to only a small section of their analysed genomes reflect collectively important uncertainties in this study. More in-depth and extensive investigations are required in order to gain greater insight into the rabies transmission risk pathways for cattle and other agricultural animals in Georgia.

Title of work

Animal Disease Early Warning System through the One Health Perspective

Co-author(s)

Giancarlo Belluzzi, Angelo Ferrari, Daniela Meloni

Introduction

Since 2006, the tripartite international organisation FAO-OIE-WHO has been establishing a Global Early Warning System for Major Animal Diseases at international level, which includes Zoonosis (GLEWS), which evolved into GLEWS+. The system additionally conducts timely robust joint risk assessments, the aim of which is to formulate risk management options for health events at the human-animal-ecosystems interface. There is no functional GLEWS+ without a well-developed Animal Disease Early Warning System at the national level. The latter could be defined as: ‘The provision of timely and effective information on exceptional disease events detected early in animals that allows all veterinary services to respond rapidly and effectively.’ The One Health approach, which is strongly promoted by OIE, FAO and WHO, facilitates a well-established system that is important for the early detection, prevention, control and eradication of animal disease. Such a system makes it possible to also follow the introduction of new diseases step by step as a consequence of two determining factors –climate changes and the displacement of numerous human populations from one geographical area to another.

Methodology

The multidisciplinary, integrated and holistic approach of One Health is important for the Animal Disease Early Warning System and its disease reporting component at the national level. According to an OIE survey of all 178 national delegates in 2012, disease reporting and food safety were high-priority programme areas for One Health approaches. Therefore, the authors explore published experiences on how the One Health approach impacts the Animal Disease Early Warning System. Social science tools such as situational analysis, benchmarking and best practices, which can be used by veterinary decision-makers, explore the points of reference and application of the One Health approach. Furthermore, these tools contribute to the development of an appropriate national strategy and action plan of the Animal Disease Early Warning System incorporating One Heath in the event of climate changes or shifts in new determinants.

Results

The joint activities of the animal or human health professionals have contributed to prioritisation of the disease, better understanding of the critical role of zoonotic diseases, and integration of epidemiological and laboratory data pertaining to different disciplines. Advocacy efforts to make One Health mainstream have built up capacity and contributed to the transfer of knowledge and have raised the level of awareness of national experts from various disciplines, including from central level to local level. The development of institutional structures and a framework for the implementation of One Health has supported the flexibility of joint actions, better engagement and mobilisation of the resources, and formalisation of the commitment between the parties to work together. The critical role of leadership is important in the overall multi-sectoral coordination mechanism. 
However, simply focusing on animal or human health without addressing the problem encountered by wildlife populations and/or the environment impedes a well-functioning system. Furthermore, the social science professionals, such as anthropologists, human behavioural scientists and sociologists, need to be invited to play a role in the system interface.

Discussion

The coronavirus disease (COVID-19) pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), first reported in December 2019 and thought to have emerged from an animal source and then spilled over to the human population, brings strengthening of the Animal Disease Early Warning system at national level to the top of the international agenda. Strengthening the Animal Disease Early Warning System can be achieved through institutionalisation of the One Health approach. The strategy, action plan, evaluation and indicators of the Animal Disease Early Warning System incorporating the One Health approach need to be actively developed by the national stakeholders to guarantee efficiency, effectiveness and sustainability. Communication about One Health is important as everyone shares an interest in the specific fields of action in daily life, even professionally and/or socially. All citizens become aware of and responsible for emerging and re-emerging disease events at national and international level.

Title of work

Investigating the health risk-benefit balance of transitioning to a sustainable diet in the Danish adult population

Co-author(s)

Aleksandra Davydova, Hernan Gomez Hernan,  Marianne Uhre Jakobsen, Kirsten Pilegaard,  Sayantan Sengupta, Morten Poulsen 

Introduction

Countries are updating dietary guidelines to include sustainable food patterns. Few studies provide evidence on the health impact of a population’s change from current to sustainable dietary patterns. In addition, the food safety implications of such a transition have yet to be investigated. The aim of our study was to evaluate the risk-benefit balance in terms of DALY of transition to a sustainable diet in Denmark, taking dietary factors and exposure to chemical contaminants into account in the estimation.

Methodology

We assessed published studies to identify intakes of food complying with various sustainability indicators. For identified food groups, we selected the health effects with at least a probable association between food and disease. We identified chemical contaminants relevant for each food group and their associated adverse effects. Contaminants were selected if a change in exposure from the current to the sustainable intake was expected. For dietary factors, a comparative risk assessment approach was applied for estimating the changes in incidences due to changes in intake from the current to the sustainable food, by combining relative risk functions with population intake assessment. For the chemical contaminants, quantitative chemical risk assessments were applied, combining dose response relations with exposure assessments to generate disease incidences at current and sustainable intake levels. Changes in DALYs due to the transition were derived by means of the total DALY for included health effects from the Global Burden of Disease project. Intake data was obtained from The Danish National Survey on Diet and Physical Activity and Danish concentration data on contaminants.

Results

As a sustainable diet scenario for the Danish population, we selected a model based on the EAT Lancet Planetary health diet, but adjusted for current Danish dietary patterns. We included nine food groups and, based on grading of evidence, we included the following diseases across food groups: type II diabetes, coronary heart disease, colorectal cancer, stroke and hypertension. We accounted for the exposure to inorganic arsenic, methyl mercury, lead, cadmium, aflatoxin and lectins across the food groups, and the associated health effects: lung, bladder, skin and liver cancer, chronic kidney disease, coronary heart disease and neurodevelopmental toxicity (loss of IQ). For the Danish population, we estimated a mean change in intake from the current to the sustainable diet scenario of 100 %, 36 %, 58 %, -26 %, -70 %, 32 %, 3900 %, 667 % and -55 % for cereals, vegetables, fruits, dairy, red meat, fish, pulses, nuts and added sugar, respectively. Final estimates are currently being prepared on the change in chemical exposures and the overall DALY difference between current and sustainable diets accounting for all included health effects.

Discussion

To our knowledge, this is the first study to quantitatively assess the risk-benefit balance of a transition from current diets to a sustainable diet, covering both the nutritional and sustainability aspects as well as food safety. As a sustainable diet, we defined a scenario based on the EAT Lancet diet but adjusted to current Danish dietary patterns. A sustainable diet can be formulated in various other ways, taking into account different indicators or reflecting different dietary patterns. However, the models developed in the project can be adapted to any definition of sustainable diets applied in other national or regional settings. We argue that the quantitative evidence generated from our study provides decision-makers with information on the trade-offs involved in the transition to a sustainably healthy diet. The quantitative assessment of the health impact provides the basis for including other indicators of public health burdens, e.g. cost of illness estimates, thus enabling the development of guidelines and interventions based on holistic scientific evidence.

Title of work

A Convention on Animal Protection to Prevent Future Pandemics

Co-author(s)

Nigel Blackaby, Nicolas Cordoba, Rosario Galardi, David Favre, Daina Bray, Rajesh Reddy, Joan Schaffner

Introduction

The COVID-19 pandemic has drawn a renewed focus to an existing gap in international law: the lack of international standards for human interactions with animals and the environment. Our team’s work seeks to address environmental law, animal well-being and the root causes of zoonotic diseases by drafting and promoting the adoption of a Convention on Animal Protection (CAP). The CAP project touches upon all subjects within the One Planet theme of this conference, and animal welfare and the search for sustainability in particular. 
As humans and domesticated animals continue to come into closer contact with wild animals as population growth, climate change and deforestation endure, the likelihood of viruses from wild animals directly or indirectly coming into contact with humans increases. Given the growing potential for the emergence of additional pandemics and the heightened public interest in these issues as a result of the COVID-19 pandemic, now is the time to address the public health risks posed by environmental degradation and the inappropriate treatment of animals.

Methodology

The CAP proposes an international regulatory framework based on case studies and research on the causes of past pandemics, epidemics and the general spillover of zoonotic diseases from animals to humans. COVID-19 is only the latest example of such a spillover. Others include AIDS, Severe Acute Respiratory Syndrome (SARS) and Ebola. Nearly all spillovers have resulted from areas that lack strong international regulations, namely the wildlife trade, inappropriate mixing of livestock and wild animals, treatment of animals in live animal markets or medical testing facilities, and destruction of natural habitats. 

After better understanding the causes of pandemics, our team engaged in legal research to understand which regulatory frameworks are most likely to be effective in preventing spillover. This process considered both (1) which regulations, if accepted, would best mitigate the risk of spillover and (2) which regulations are most likely to gain traction on a global scale. 

Our team is currently engaging in outreach with senior officials in government, non-governmental organisations and academia to promote the CAP and obtain feedback on the draft treaty itself.

Results

The result of our research is the Convention on Animal Protection. The CAP contains four parts: an introduction, substantive provisions, provisions on the functioning of the treaty, and an annex identifying species with a high-risk of transmitting zoonotic diseases and other pathogens to humans and other animals. 
The CAP contemplates the adoption of the first sets of international minimum standards of care for animals, including obligations to provide animals with suitable and sufficient food and water, adequate shelter from adverse environmental conditions and adequate opportunities for exercise and mental stimulation.

In particular, the CAP provides regulations on human interactions with animals in several broad categories: all animals; animals deemed to have a high risk of hosting viruses or other pathogens that pose a serious risk to public or animal health; companion animals; commercial animals; animals used in scientific research and testing; and animals used in entertainment.

Discussion

If implemented, the CAP would be the first binding international legal instrument (treaty) to incorporate the One Health principle. It would also be the first treaty to address the root causes of pandemics and epidemics: the unnecessary spread of zoonotic viruses and other pathogens. Other treaties and initiatives under discussion tend to be reactive, in that they focus more on how better to react to the next pandemic, whereas this treaty is proactive, in seeking to make pandemics significantly less likely events. 

As it is currently drafted, the CAP is a framework for further discussion on practical, binding ways to implement the concept of One Health, environment and society. As was the case for the negotiation of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES) (and many other treaties on various topics that began as projects by independent groups), our team has taken it upon ourselves to write the first draft of the treaty. Now, we are looking for state policymakers to pick up the pen. 

The science is clear – the spread of zoonotic viruses is preventable. Now, we need policymakers to take action.

Title of work

Detection of a carbapenem-resistant Klebsiella pneumoniae strain in kitchen sponges

Co-author(s)

Laurence Delbrassinne, Hanne Debergh, Cristina Garcia-Graells, Koenraad Van Hoorde

Introduction

Although research on foodborne diseases is well documented for certain micro-organisms, less research has been undertaken on cross-contamination with Enterobacteriaceae from food within the kitchen environment. In this study, we analysed the presence of Klebsiella spp. and Raoultella spp. and their antibiotic resistance in 100 used kitchen sponges. In a domestic environment, kitchen sponges are commonly used by consumers for doing the dishes and/or cleaning kitchen surfaces. Given their multi-purpose use, their often high humidity and the presence of organic residuals, they are considered a favourable habitat for various groups of micro-organisms and could serve as a vehicle in the transmission of foodborne pathogens.

Methodology

A total of 100 kitchen sponges, alongside a questionnaire regarding hygienic parameters, were randomly collected from domestic environments and analysed within 24 hours after arrival in the lab. Kitchen sponges were immersed in 100 ml buffered peptone water, homogenised and incubated at 37°C for 24 h ± 2 h. Ten µl of this enrichment was plated on MacConkey agar and incubated at 37°C for 24 h ± 2 h. Presumptive Klebsiella spp. and Raoultella spp. were isolated and confirmed by MALDI-TOF MS. Antibiotic susceptibility testing (AST) was performed on all Klebsiella spp. and Raoultella spp. isolates following the EUCAST guidelines using EUVSEC 3 and EUVSEC 2 plates. Whole genome sequencing was performed using the Illumina platform.

Results

A total of 60 % of the kitchen sponges were positive for Klebsiella spp. or Raoultella spp and six sponges contained two different species. Klebsiella oxytoca was detected in 78.5 % of the positive samples, followed by Klebsiella pneumoniae (12.31 %), Raoultella ornitholytica (4.62 %), Klebsiella variicola (1.54 %), Klebsiella aerogenes (1.54 %) and Raoultella planticola (1.54 %). AST revealed the presence of one carbapenem-resistant isolate (ST26), with resistance to ertapenem and meropenem. Reduced susceptibility to carbapenems was caused by the presence of blaSHV-36 combined with porin deficiency (OmpK37).

Discussion

Klebsiella spp. can be part of the commensal flora of human intestines but are also considered to be opportunistic pathogens. Klebsiella spp. might present antimicrobial resistance, and carbapenem-resistant Enterobacteriaceae are included in the WHO global priority list of antibiotic-resistant bacteria. 
Among the Klebsiella spp. isolates, K. oxytoca was the most prevalent species (52/57, 91.23 %) followed by K. pneumoniae (8/57, 14.04 %), as previously observed (Marotta et al.; Møretrø et al.; Osaili et al.). This high prevalence of Klebsiella spp. as well as the presence of a carbapenem-resistant isolate in kitchen sponges raise concern about the possible risks of foodborne diseases associated with this kitchen tool. These findings advocate for good hygienic measures within a household setting.

Title of work

A comparison of two different approaches to investigate the role of air temperature as a key driver of salmonellosis

Co-author(s)

Alasdair Cook, Caitriona Fenton, Emma Gillingham, Theo Kanellos, Gordon Nichols, Joaquin M. Prada, Giovanni Lo Iacono

Introduction

Salmonellosis continues to be a major cause of disease in humans and animals, being the first agent involved in foodborne outbreaks in Europe. The observed incidence of salmonellosis in humans reveals a seasonal pattern, with a higher number of cases reported during the warmest months. This seasonality alludes to the importance of the environment as a modulator of infection. The main driver of seasonality is believed to be temperature, but other influential variables may well be involved. Our goal is to explore the role of temperature as a key weather variable driving the incidence of salmonellosis in humans. We want to test the hypothesis that seasonality in the incidence of salmonellosis is largely driven by the response of bacterial growth in food to air temperature.

Methodology

We will compare the outcomes of two different approaches, using temperature records for both. The first method is based on a mechanist approach that simulates the number of salmonellosis cases due to the growth of temperature-driven Salmonella in eggs. We simulated the number of human salmonellosis cases observed during the past 30 years at a postcode-resolution location in England and Wales with a mechanistic model and using historical temperatures as input. Our estimations were based on a Poisson process with a rate of infection assumed to be proportional to the bacterial load in relevant food (i.e. eggs and chicken). The bacterial load, in turn, depends on air temperature according to the empirical relationship available in the literature. On the basis of the mechanistic model, we estimated the theoretical probability of observing a case of salmonellosis at a given temperature. The second method is a statistical approach, where we used the disease records and their spatio-temporal links to environmental variables (temperature, precipitation, humidity, vapour pressure, and UV radiation) to estimate the empirical probability of finding a case of salmonellosis conditional to a range of temperatures.

Results

In a subsequent step, we will compare simulated and observed conditional probabilities to validate the underlying hypothesis. With the first approach, we have found a positive agreement between the predicted and observed incidence curves for England and Wales. This tentatively confirms the relevant role that temperature and eggs play in salmonellosis temporal incidence patterns – positioning temperature as a potential primary weather driver of salmonellosis. We will validate this finding if both approaches lead to a similar conditional incidence of salmonellosis to temperature. On the other hand, if the results do not match, this would indicate a research gap and point to further experiments involving other food sources of salmonellosis as well as other weather variables involved.

Discussion

Having solid proof of the main environmental factors that influence salmonellosis transmission is important to tackle future disease events, especially in the framework of global warming. Understanding why the incidence of salmonellosis is conditioned to certain weather variables will be useful for practical public health applications to better elucidate observed spatio-temporal patterns and improve future incidence predictions. In particular, we will build on this model to develop a tool to predict the likelihood of infection based on known weather variations prior to the occurrence of an infection. Once the model has been validated, it is our aim to apply it to a different geographic area to test the efficacy of the model.

Title of work

The COMPARE Database: A Comprehensive Public Resource for Allergen Identification and Protein Allergenicity Assessment

Co-author(s)

Laurent Beuf, Gabriele Gadermaier, Zhongshan Gao, Eva Gietl, R Karin, Liisa Koski, Justin McDonald, Clare Narrod, Elda Posada-Campos, Andre Silvanovich, Ping Song, Wiebke Striegel, Suzanne Teuber, Ronald van REE, Lucilia Pereira Mouriès

Introduction

Protein allergenicity is associated with specific protein structure and amino acid sequences that bind immunoglobulin E, as opposed to being linked to protein function. Therefore, a sequence database composed of validated allergens is seen as essential for the safety assessment of genetically modified foods and feeds, as well as other innovative food/feed products. However, the identification of new allergen sequences has become more complex due to the exponential increase of sequence data that accompanied the broad adoption of high throughput sequencing methods.

Methodology

To address these challenges, an international collaborative scientific group coordinated by the Health and Environmental Sciences Institute (HESI) was tasked to develop a contemporary, adaptable, high-throughput process to build the COMprehensive Protein Allergen REsource (COMPARE) database. The COMPARE database is updated annually using a transparent, computer-based process to identify candidate sequences in scientific literature and publicly accessible protein sequence databases. Candidate sequences are then reviewed and approved by an independent international panel of academic and clinical allergy experts for inclusion in COMPARE.

Results

The COMPARE database, together with its built-in bioinformatic sequence search and visualisation tool known as COMPASS, and extensive documentation, are publicly available at https://comparedatabase.org/. The incorporation of the FASTA sequence similarity search algorithm as the core tool in the COMPARE database supports real-time bioinformatic comparisons using global allergen sequence similarity metrics that are based on FAO/WHO and CODEX Alimentarius guidelines. The entire list of sequences and accompanying metadata can be downloaded from the website.

Discussion

This presentation will highlight the most recent database update cycle culminating in the January 2022 COMPARE release.
Altogether, the COMPARE database and its publicly available resources support the development of innovative food products and technologies by providing up-to-date tools for allergenicity assessment before new products enter the market, and thus contribute to better protection of human health and food safety.