TKPlate 1.0: An Open-access platform for toxicokinetic and toxicodynamic modelling of chemicals to implement new approach methodologies in chemical risk assessment.

This publication is linked to the following EFSA Supporting Publications articles: http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2023.EN-8441/full, http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2023.EN-8440/full, http://onlinelibrary.wiley.com/doi/10.2903/sp.efsa.2023.EN-8437/full.

Lumpy skin disease epidemiological report IV: data collection and analysis.

In 2019, no lumpy skin disease (LSD) outbreaks were reported in South-Eastern Europe, the mass vaccination regional campaign with homologous LSD vaccine continued for the fourth year with over 1.8 million bovines vaccinated in the region, preventing further outbreaks since 2016. LSD outbreaks were reported in Turkey, including western Turkey, in Russia and in eastern Asia affecting China, Bangladesh and India for the first time. The use of homologous vaccine should be considered in the countries still affected in order to eliminate the virus. Besides passive surveillance, which is implemented in all the countries, active surveillance for early detection based on clinical examination could be conducted ideally during April-October every 5 weeks in at-risk areas, based on possible re-emergence or re-introduction from affected neighbouring countries. Active surveillance for proving disease freedom could be based on serological testing (enzyme-linked immunosorbent assay (ELISA)) targeting 3.5% seroprevalence and conducted on a random sample of cattle herds on non-vaccinated animals. LSD re-emerged in Israel in 2019, after vaccination became voluntary. This shows that, if the virus is still circulating in the region, the reduced protection might result in re-emergence of LSD. In case of re-emergence, a contingency plan and vaccine stockpiling would be needed, in order to react quickly. From a study performed in Israel to test side effects of live-attenuated homologous LSD vaccine, milk production can be reduced during 7 days after vaccination (around 6-8 kg per cow), without a significant loss in the 30 days after vaccination. Research needs should be focused on the probability of transmission from insect to bovine, the virus inactivation rate in insects, the collection of baseline entomological data, the capacity of vector species in LSDV transmission linked to studies on their abundance and the control of Stomoxys calcitrans being the most important vector in LSD transmission.

Occurrence and Co-Occurrence of Mycotoxins in Cereal-Based Feed and Food.

Dietary (co)-exposure to mycotoxins is associated with human and animal health concerns as well as economic losses. This study aims to give a data-based insight from the scientific literature on the (co-)occurrence of mycotoxins (i.e., parent and modified forms) in European core cereals, and to estimate potential patterns of co-exposure in humans and animals. Mycotoxins were mainly reported in wheat and maize showing the highest concentrations of fumonisins (FBs), deoxynivalenol (DON), aflatoxins (AFs), and zearalenone (ZEN). The maximum concentrations of FB1+FB2 were reported in maize both in feed and food and were above legal maximum levels (MLs). Similar results were observed in DON-food, whose max concentrations in wheat, barley, maize, and oat exceeded the MLs. Co-occurrence was reported in 54.9% of total records, meaning that they were co-contaminated with at least two mycotoxins. In the context of parental mycotoxins, co-occurrence of DON was frequently observed with FBs in maize and ZEN in wheat; DON + NIV and DON + T2/HT2 were frequently reported in barley and oat, respectively. Apart from the occurrence of ZEN and its phase I and phase II modified forms, only a limited number of quantified data were available for other modified forms; i.e., mainly the acetyl derivatives of DON. Data gaps are highlighted together with the need for monitoring studies on multiple mycotoxins to identify co-occurrence patterns for parent mycotoxins, metabolites, and their modified forms.

Inferences about the transmission of lumpy skin disease virus between herds from outbreaks in Albania in 2016.

Lumpy skin disease has recently emerged as a major threat to cattle populations outside of Africa, where it is endemic. In 2015 the first ever European outbreaks occurred in Greece, which were followed by spread across much of the Balkans in 2016. Here we use a simple mathematical model for the transmission of lumpy skin disease virus (LSDV) between herds to explore factors influencing its spread by fitting it to data on outbreaks in Albania in 2016. We show that most transmission occurs over short distances (<5 km), but with an appreciable probability of transmission at longer distances. We also show that there is evidence for seasonal variation in the force of infection associated with temperature, possibly through its influence on the relative abundance of the stable fly, Stomoxys calcitrans. These two results together are consistent with LSDV being transmitted by the bites of blood-feeding insects, though further work is required to incriminate specific species as vectors. Finally, we show that vaccination has a significant impact on spread and estimate the vaccine effectiveness to be 76%.

Investigating combined toxicity of binary mixtures in bees: Meta-analysis of laboratory tests, modelling, mechanistic basis and implications for risk assessment.

Bees are exposed to a wide range of multiple chemicals "chemical mixtures" from anthropogenic (e.g. plant protection products or veterinary products) or natural origin (e.g. mycotoxins, plant toxins). Quantifying the relative impact of multiple chemicals on bee health compared with other environmental stressors (e.g. varroa, viruses, and nutrition) has been identified as a priority to support the development of holistic risk assessment methods. Here, extensive literature searches and data collection of available laboratory studies on combined toxicity data for binary mixtures of pesticides and non-chemical stressors has been performed for honey bees (Apis mellifera), wild bees (Bombus spp.) and solitary bee species (Osmia spp.). From 957 screened publications, 14 publications provided 218 binary mixture toxicity data mostly for acute mortality (lethal dose: LD50) after contact exposure (61%), with fewer studies reporting chronic oral toxicity (20%) and acute oral LC50 values (19%). From the data collection, available dose response data for 92 binary mixtures were modelled using a Toxic Unit (TU) approach and the MIXTOX modelling tool to test assumptions of combined toxicity i.e. concentration addition (CA), and interactions (i.e. synergism, antagonism). The magnitude of interactions was quantified as the Model Deviation Ratio (MDR). The CA model applied to 17% of cases while synergism and antagonism were observed for 72% (MDR > 1.25) and 11% (MDR < 0.83) respectively. Most synergistic effects (55%) were observed as interactions between sterol-biosynthesis-inhibiting (SBI) fungicides and insecticide/acaricide. The mechanisms behind such synergistic effects of binary mixtures in bees are known to involve direct cytochrome P450 (CYP) inhibition, resulting in an increase in internal dose and toxicity of the binary mixture. Moreover, bees are known to have the lowest number of CYP copies and other detoxification enzymes in the insect kingdom. In the light of these findings, occurrence of these binary mixtures in relevant crops (frequency and concentrations) would need to be investigated. Addressing this exposure dimension remains critical to characterise the likelihood and plausibility of such interactions to occur under field realistic conditions. Finally, data gaps and further work for the development of risk assessment methods to assess multiple stressors in bees including chemicals and non-chemical stressors in bees are discussed.

Testing the thresholds of toxicological concern values using a new database for food-related substances.

The Threshold of Toxicological Concern (TTC) concept integrates data on exposure, chemical structure, toxicity and metabolism to identify a safe exposure threshold value for chemicals with insufficient toxicity data for risk assessment. The TTC values were originally derived from a non-cancer dataset of 613 compounds with a potentially small domain of applicability. There is interest to test whether the TTC values are applicable to a broader range of substances, particularly relevant to food safety using EFSA's new OpenFoodTox database. After exclusion of genotoxic compounds, organophosphates or carbamates or those belonging to the TTC exclusion categories, the remaining 329 substances in the EFSA OpenFoodTox database were categorized under the Cramer decision tree, into low (Class I), moderate (II), or high (III) toxicity profile. For Cramer Classes I and III the threshold values were 1000 µg/person per day (90% confidence interval: 187-2190) and 87 µg/person per day (90% confidence interval: 60-153), respectively, compared to the corresponding original threshold values of 1800 and 90 µg/person per day. This confirms the applicability of the TTC values to substances relevant to food safety. Cramer Class II was excluded from our analysis because of containing too few compounds. Comparison with the Globally Harmonized System of classification confirmed that the Cramer classification scheme in the TTC approach is conservative for substances relevant to food safety.

Risk assessment of African swine fever in the south-eastern countries of Europe.

The European Commission requested EFSA to estimate the risk of spread of African swine fever (ASF) and to identify potential risk factors (indicators) for the spread of ASF, given introduction in the south-eastern countries of Europe (region of concern, ROC), namely Albania, Bosnia and Herzegovina, Croatia, Greece, Kosovo, Montenegro, North Macedonia, Serbia and Slovenia. Three EU Member States (MS) - Croatia, Greece and Slovenia - were included in the ROC due to their geographical location and ASF-free status. Based on collected information on potential risk factors (indicators) for each country and the relevant EU regulations in force, the estimated probability of spread of ASF within the ROC within one year after introduction into the ROC was assessed to be very high (from 66% to 100%). This estimate was determined after considering the high number of indicators present in most of the countries in the ROC and the known effect that these indicators can have on ASF spread, especially those related to the structure of the domestic pig sector, the presence of wild boar and social factors. The presence of indicators varies between countries in the ROC. Each country is at risk of ASF spread following introduction; however, some countries may have a higher probability of ASF spread following introduction. In addition, the probability of ASF spread from the ROC to EU MSs outside the ROC within one year after introduction of ASF in the ROC was estimated to be very low to low (from 0% to 15%). This estimate was based on the comparison of the indicators present in the ROC and the already affected countries in south-eastern Europe, such as Bulgaria and Romania, where there was no evidence of ASF spread to other EU MS within one year.

Lumpy skin disease: III. Data collection and analysis.

In 2018, no lumpy skin disease (LSD) outbreaks were reported in the Balkan region, after the decline reported in 2017 (385) compared to 2016 (7,483). This confirms the effectiveness of the vaccination campaign based on the LSD homologous vaccine strain which continued throughout 2018 with over 2.5 million animals vaccinated, keeping the mean vaccination coverage above 70%. In 2018, LSD outbreaks were reported in Russia, Turkey and Georgia. In Russia, the LSD epidemics expanded northward and eastward, while in Turkey, the most affected region was in the east. LSD is spreading in Turkey since 2013, despite large vaccination campaigns with heterologous vaccine performed since 2014. This might support the hypothesis that the use of heterologous vaccines results in insufficient protection, and therefore, the use of homologous LSD vaccine in Turkey should be considered to prevent further spread. As the LSD epidemic in Turkey is a risk for reintroduction into the EU, it is recommended to continue the vaccination campaigns in 2019 in the high-risk areas of Balkan region. Spread rates of LSD within a village were estimated from outbreak data for Albania, which can be used to inform the level of vaccination required to control an outbreak in a village. In terms of vaccine safety, the reports from the field suggest that, compared to the large number of animals vaccinated in the Balkan region since 2015, a very limited number of side effects have been recorded so far, although from published literature, local or even systemic side effects in some animals may occur after vaccination. However, due to inadequate study design in the reviewed studies, there is no consensus on the magnitude of such effects and on their real consequences on production.

Research gap analysis on African swine fever.

The most significant knowledge gaps in the prevention and control of African swine fever (ASF) were identified by the EU Veterinary services and other stakeholders involved in pig production and wild boar management through an online survey. The respondents were asked to identify the major research needs in order to improve short-term ASF risk management. Four major gaps were identified: 'wild boar', 'African swine fever virus (ASFV) survival and transmission', 'biosecurity' and 'surveillance'. In particular, the respondents stressed the need for better knowledge on wild boar management and surveillance, and improved knowledge on the possible mechanism for spread and persistence of ASF in wild boar populations. They indicated the need for research on ASFV survival and transmission from the environment, different products such as feed and feed materials, and potential arthropod vector transmission. In addition, several research topics on biosecurity were identified as significant knowledge gaps and the need to identify risk factors for ASFV entry into domestic pig holdings, to develop protocols to implement specific and appropriate biosecurity measures, and to improve the knowledge about the domestic pig-wild boar interface. Potential sources of ASFV introduction into unaffected countries need to be better understood by an in-depth analysis of the possible pathways of introduction of ASFV with the focus on food, feed, transport of live wild boars and human movements. Finally, research on communication methods to increase awareness among all players involved in the epidemiology of ASF (including truck drivers, hunters and tourists) and to increase compliance with existing control measures was also a topic mentioned by all stakeholders.

Lumpy skin disease: scientific and technical assistance on control and surveillance activities.

The duration of the vaccination campaign sufficient to eliminate lumpy skin disease (LSD) mainly depends on the vaccination effectiveness and coverage achieved. By using a spread epidemiological model, assuming a vaccination effectiveness of 65%, with 50% and 90% coverage, 4 and 3 years campaigns, respectively, are needed to eliminate LSD. When vaccination effectiveness is 80% to 95%, 2 years of vaccination at coverage of 90% is sufficient to eliminate LSD virus (LSDV). For shorter campaigns, LSD is predicted to persist. When the infection is eliminated by vaccination, two pathways for disease recurrence are possible, (i) by new introduction from a neighbouring affected area, especially by introduction of infected animals, or, less likely (ii) the infection persisting either in the environment, in vectors or in wild animals. For planning surveillance, several elements should be considered: the objectives and related design prevalence, the epidemiological situation, the immunological status of the host population, the geographical area and the season, the type of surveillance (active or passive), the diagnostic methods including clinical detection (considered the most effective method for early detection of LSD), the target population, the sample size and frequency. According to the model, for early detecting new introductions of LSD, it may be needed to clinically check a large number of herds (e.g. 2-3,000 herds) monthly. Lower sample sizes can be considered, when a greater delay in detecting the virus is acceptable. Where vaccination is maintained, active surveillance for verifying the effectiveness of vaccination would be needed. Demonstrating disease absence can rely on serological surveillance, which should consider the test sensitivity, the design prevalence (estimated value: 3.5%), the onset and duration of serum antibodies. Important knowledge gaps on LSD are about within-herd transmission, duration of protective immunity, role of vectors, diagnostic tests, farm location and type in the at-risk countries and the epidemiological status of neighbouring countries.