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.

Epidemiological analyses of African swine fever in the European Union (November 2017 until November 2018).

This update on the African swine fever (ASF) outbreaks in the EU demonstrated that out of all tested wild boar found dead, the proportion of positive samples peaked in winter and summer. For domestic pigs only, a summer peak was evident. Despite the existence of several plausible factors that could result in the observed seasonality, there is no evidence to prove causality. Wild boar density was the most influential risk factor for the occurrence of ASF in wild boar. In the vast majority of introductions in domestic pig holdings, direct contact with infected domestic pigs or wild boar was excluded as the route of introduction. The implementation of emergency measures in the wild boar management zones following a focal ASF introduction was evaluated. As a sole control strategy, intensive hunting around the buffer area might not always be sufficient to eradicate ASF. However, the probability of eradication success is increased after adding quick and safe carcass removal. A wider buffer area leads to a higher success probability; however it implies a larger intensive hunting area and the need for more animals to be hunted. If carcass removal and intensive hunting are effectively implemented, fencing is more useful for delineating zones, rather than adding substantially to control efficacy. However, segments of fencing will be particularly useful in those areas where carcass removal or intensive hunting is difficult to implement. It was not possible to demonstrate an effect of natural barriers on ASF spread. Human-mediated translocation may override any effect of natural barriers. Recommendations for ASF control in four different epidemiological scenarios are presented.

Effect on public health of a possible increase of the maximum level for 'aflatoxin total' from 4 to 10 µg/kg in peanuts and processed products thereof, intended for direct human consumption or use as an ingredient in foodstuffs.

EFSA was asked to deliver a scientific opinion regarding the effect on public health of a possible increase of the maximum level (ML) for 'aflatoxin total' (AFT; sum of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2) from 4 to 10 µg/kg in peanuts and processed products thereof. Aflatoxins are genotoxic and cause hepatocellular carcinomas in humans. The Panel on Contaminants in the Food Chain (CONTAM Panel) evaluated 8,085 samples of peanuts and 472 samples of peanut butter, with > 60% left-censored. The mean concentration of AFT in peanuts was 2.65/3.56 µg/kg (lower bound (LB)/upper bound (UB)) with a maximum of 1,429 µg/kg. The mean concentration in peanut butter was 1.47/1.92 µg/kg (LB/UB) with a maximum of 407 µg/kg. Peanut oil was not included since all data were left-censored and the ML does not apply for oil. Exposure was calculated for a 'Current ML' and 'Increased ML' scenario, and mean chronic exposure estimates for consumers only, amounted to 0.04-2.74 ng/kg body weight (bw) per day and 0.07-4.28 ng/kg bw per day, respectively. The highest exposures were calculated for adolescents and other children. The CONTAM Panel used the cancer potencies estimated by the Joint FAO/WHO Expert Committee on Food Additives for the risk characterisation. Under the scenario of the current ML, the cancer risk was estimated to range between 0.001 and 0.213 aflatoxin-induced cancers per 100,000 person years. Under the scenario of the increased ML, it ranged between 0.001 and 0.333 aflatoxin-induced cancers per 100,000 person years. Comparing these data calculated under the current ML scenario with the yearly excess cancer risk of 0.014 shows a higher risk for consumers of peanuts and peanut butter in some surveys. The calculated cancer risks indicate that an increase of the ML would further increase the risk by a factor of 1.6-1.8.

African swine fever in wild boar.

The European Commission requested EFSA to compare the reliability of wild boar density estimates across the EU and to provide guidance to improve data collection methods. Currently, the only EU-wide available data are hunting data. Their collection methods should be harmonised to be comparable and to improve predictive models for wild boar density. These models could be validated by more precise density data, collected at local level e.g. by camera trapping. Based on practical and theoretical considerations, it is currently not possible to establish wild boar density thresholds that do not allow sustaining African swine fever (ASF). There are many drivers determining if ASF can be sustained or not, including heterogeneous population structures and human-mediated spread and there are still unknowns on the importance of different transmission modes in the epidemiology. Based on extensive literature reviews and observations from affected Member States, the efficacy of different wild boar population reduction and separation methods is evaluated. Different wild boar management strategies at different stages of the epidemic are suggested. Preventive measures to reduce and stabilise wild boar density, before ASF introduction, will be beneficial both in reducing the probability of exposure of the population to ASF and the efforts needed for potential emergency actions (i.e. less carcass removal) if an ASF incursion were to occur. Passive surveillance is the most effective and efficient method of surveillance for early detection of ASF in free areas. Following focal ASF introduction, the wild boar populations should be kept undisturbed for a short period (e.g. hunting ban on all species, leave crops unharvested to provide food and shelter within the affected area) and drastic reduction of the wild boar population may be performed only ahead of the ASF advance front, in the free populations. Following the decline in the epidemic, as demonstrated through passive surveillance, active population management should be reconsidered.

Thermal processing of live bivalve molluscs for controlling viruses: On the need for a risk-based design.

Norovirus (NoV) and Hepatitis A virus (HAV) are the most important viral hazards associated with human illness following consumption of contaminated bivalve molluscs. The effectiveness of the current EU criteria for heat processing of bivalve molluscs (i.e. raising the temperature of the internal mollusc flesh to at least 90°C for a minimum of 90 seconds) was evaluated using predictive microbiology. A HAV thermal inactivation model was developed based on literature data in mollusc matrices during isothermal heat treatment. Application of the developed model demonstrated that the 90°C-90 s requirement may lead to significantly different virus inactivation depending on the commercial process design. This shows the need for the establishment of a Performance Criterion for bivalve molluscs heat processing which will assure a common specified level of consumer protection. A risk-based approach is described that allows for an effective processing design providing a more transparent and objective relation between the thermal processing targets and public health. Model simulations demonstrate that the F-value is a more appropriate Process Criterion than a single time-temperature combination since it enables the food business operators to design a process that is compliant with the safety requirements while at the same time achieving a desired product quality.

One Health Economics to confront disease threats.

Global economic impacts of epidemics suggest high return on investment in prevention and One Health capacity. However, such investments remain limited, contributing to persistent endemic diseases and vulnerability to emerging ones. An interdisciplinary workshop explored methods for country-level analysis of added value of One Health approaches to disease control. Key recommendations include: 1. systems thinking to identify risks and mitigation options for decision-making under uncertainty; 2. multisectoral economic impact assessment to identify wider relevance and possible resource-sharing, and 3. consistent integration of environmental considerations. Economic analysis offers a congruent measure of value complementing diverse impact metrics among sectors and contexts.

Epidemiological analyses of African swine fever in the Baltic States and Poland: (Update September 2016-September 2017).

EFSA assisted four countries in the analysis of epidemiological data on African swine fever (ASF), collected until September 2017. The temporal analysis demonstrated that the average proportions of PCR and antibody-ELISA positive samples from the hunted wild boar remained below 3.9 and 6.6, respectively. A peak in the ASF incidence was observed 6 months after the first observed case, followed by a significant reduction of the number of cases and low levels of African swine fever virus (ASFV) circulation at the end of 38 months follow-up period at different spatial resolutions. The spatial analysis concluded that human-mediated spread of ASFV continues to play a critical role in the ASF epidemiology, despite all measures currently taken. 'Wild boar density', 'total road length' (as proxy for human activity) and 'average suitable wild boar habitat availability' were identified as predictors for the occurrence of ASF in Estonia by a Bayesian hierarchical model, whereas 'wild boar density' and 'density of pig farms' were predictors according to a generalised additive model. To evaluate the preventive strategies proposed in EFSA's Scientific Opinion (2015) to stop the spread of ASFV in the wild boar population, a simulation model, building on expert knowledge and literature was used. It was concluded that reduction of wild boar population and carcass removal to stop the spread of ASFV in the wild boar population are more effective when applied preventively in the infected area. Drastic depopulation, targeted hunting of female wild boar and carcass removal solely implemented as measures to control ASF in the wild boar population need to be implemented in a highly effective manner (at or beyond the limit of reported effectivity in wild boar management) to sustainably halt the spread of ASF.

Low atmospheric pressure system for stunning broiler chickens.

Council Regulation (EC) No 1099/2009 on the protection of animals at the time of killing lists in Annex I the stunning interventions currently allowed in the EU, together with the related conditions under which those interventions can be implemented. The regulation allows the Commission to amend Annex I, listing additional stunning interventions, provided they ensure a level of animal welare at least equivalent to that ensured by the one already approved. EFSA was requested to perform such assessment with regard to the implementation of the low atmospheric pressure stunning (LAPS) system on broiler chickens. The ad hoc Working Group (WG) set up by EFSA performed the assessment in three main steps, i.e. checking the data provided against the criteria laid down in the EFSA Guidance (EFSA AHAW Panel, 2013); running an extensive literature search, followed by data extraction and performing a judgemental ranking exercise based on expert opinion. As main outcome, the LAPS intervention was found to be able to provide a level of animal welfare not lower than that provided by at least one of the currently allowed methods. The overall assessment of EFSA is valid ONLY under the technical conditions described in the submission and for broiler chickens, intended for human consumption, weighting less than 4 kg. Deviations from these conditions might have different consequences for animal welfare which were not assessed in this exercise. The LAPS method may, in addition to commercial slaughter, be suitable for depopulation, respecting the technical conditions defined in the present conclusions. The WG considers that a revision of the present version of the EFSA Guidance could be beneficial.

Epidemiological analyses on African swine fever in the Baltic countries and Poland.

African swine fever virus (ASFV) has been notified in the Baltic countries and the eastern part of Poland from the beginning of 2014 up to now. In collaboration with the ASF-affected Member States (MS), EFSA is updating the epidemiological analysis of ASF in the European Union which was carried out in 2015. For this purpose, the latest epidemiological and laboratory data were analysed in order to identify the spatial-temporal pattern of the epidemic and a risk factors facilitating its spread. Currently, the ASF outbreaks in wild boar in the Baltic countries and Poland can be defined as a small-scale epidemic with a slow average spatial spread in wild boar subpopulations (approximately from 1 in Lithuania and Poland to 2 km/month in Estonia and Latvia). The number of positive samples in hunted wild boar peaks in winter which can be explained by human activity patterns (significant hunting activity over winter). The number of positive samples in wild boar found dead peaks in summer. This could be related to the epidemiology of the disease and/or the biology of wild boar; however, this needs further investigation. Virus prevalence in hunted wild boar is very low (0.04-3%), without any apparent trend over time. Apparent virus prevalence at country level in wild boar found dead in affected countries ranges from 60% to 86%, with the exception of Poland, where values between 0.5% and 1.42%, were observed. Since the beginning of the epidemic, the apparent antibody prevalence in hunted wild boar has always been lower than the apparent virus prevalence, indicating an unchanged epidemiological/immunological situation. The risk factor analysis shows an association between the number of settlements, human and domestic pigs population size or wild boar population density and the presence of ASF in wild boar for Estonia, Latvia and Lithuania.

Re-evaluation of potassium nitrite (E 249) and sodium nitrite (E 250) as food additives.

The Panel on Food Additives and Nutrient Sources added to Food (ANS) provided a scientific opinion re-evaluating the safety of potassium nitrite (E 249) and sodium nitrite (E 250) when used as food additives. The ADIs established by the SCF (1997) and by JECFA (2002) for nitrite were 0-0.06 and 0-0.07 mg/kg bw per day, respectively. The available information did not indicate in vivo genotoxic potential for sodium and potassium nitrite. Overall, an ADI for nitrite per se could be derived from the available repeated dose toxicity studies in animals, also considering the negative carcinogenicity results. The Panel concluded that an increased methaemoglobin level, observed in human and animals, was a relevant effect for the derivation of the ADI. The Panel, using a BMD approach, derived an ADI of 0.07 mg nitrite ion/kg bw per day. The exposure to nitrite resulting from its use as food additive did not exceed this ADI for the general population, except for a slight exceedance in children at the highest percentile. The Panel assessed the endogenous formation of nitrosamines from nitrites based on the theoretical calculation of the NDMA produced upon ingestion of nitrites at the ADI and estimated a MoE > 10,000. The Panel estimated the MoE to exogenous nitrosamines in meat products to be < 10,000 in all age groups at high level exposure. Based on the results of a systematic review, it was not possible to clearly discern nitrosamines produced from the nitrite added at the authorised levels, from those found in the food matrix without addition of external nitrite. In epidemiological studies there was some evidence to link (i) dietary nitrite and gastric cancers and (ii) the combination of nitrite plus nitrate from processed meat and colorectal cancers. There was evidence to link preformed NDMA and colorectal cancers.

Exploring the association between resistance and outpatient antibiotic use expressed as DDDs or packages.

OBJECTIVES: The objective of this study was to explore the association between resistance and outpatient antibiotic use, expressed as either DDDs per 1000 inhabitants per day (DID) or packages per 1000 inhabitants per day (PID). METHODS: IMS Health data on outpatient penicillin and cephalosporin (ß-lactam) and tetracycline, macrolide, lincosamide and streptogramin (TMLS) use, aggregated at the level of the active substance (WHO version 2011) expressed as DID and PID (2000-07) were linked to European Antimicrobial Resistance Surveillance System (EARSS) data on proportions of penicillin-non-susceptible Streptococcus pneumoniae (PNSP) and erythromycin-non-susceptible S. pneumoniae (ENSP) (2000-09). Combined data for 27 European countries were analysed with a generalized linear mixed model. Model fit for use in DID, PID or both and 0, 1 or 2 year time lags between use and resistance was assessed and predictions of resistance were made for decreasing use expressed as DID, PID or both. RESULTS: When exploring the association between ß-lactam use and PNSP, the best model fit was obtained for use in PID without time lag. For the association between TMLS use and ENSP, the best model fit was obtained for use in both PID and DID with a 1 year time lag. PNSP and ENSP are predicted to decrease when use decreases in PID, but not when use decreases in DID. CONCLUSIONS: Associations between outpatient antibiotic use and resistance and predictions of resistance were inconsistent whether expressing antibiotic use as DID or PID. We recommend that data on antibiotic use be expressed as PID and that time lags between use and resistance be considered when exploring these associations.