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Background: Chronic non-cancer pain (CNCP) is one of the leading causes of disability. The use of strong opioids (SOs) in the management of CNCP is increasing, although evidence supporting their use remains limited. Primary care (PC) plays a key role in this context. Objectives: Our objectives were to determine the prevalence and profile of patients using SOs for CNCP in PC consultations in Valladolid in 2022, and to describe the consumption of SO prescribed for CNCP from 2020 to 2023. Methods: We conducted a descriptive and retrospective study using data extracted from the Pharmaceutical Consumption Information System of Castilla y León. Patients in Valladolid with SO use for more than three months due to CNCP were analyzed. Sociodemographic and clinical characteristics of these patients in 2022 were described. The number of defined daily doses (nº DDDs) and costs from 2020 to 2023 were analyzed. Results: A total of 3642 patients were included (0.7% of the population of Valladolid), 71.8% of whom were women. Of the patients, 62.4% were aged 70 or older, 39.8% lived in rural areas, and 9.9% resided in nursing homes. The most frequently prescribed SOs in nº DDDs were fentanyl and tapentadol. The highest consumption in nº DDDs was in patients who lived in nursing homes, were over 70 years old and were resident in rural areas. The number of DDDs from 2020 to 2023 for SOs in DCNO increased by 41%. Conclusions: In total, 0.7% of the population of Valladolid consumes SOs for CNCP, mostly women and people over 70 years old. The consumption of strong opioids in DDDs grew by 41% from 2020 to 2023.

The SARS-CoV-2 pandemic significantly affected the epidemiology of Streptococcus pyogenes, a pathogen associated with various clinical presentations such as pharyngitis, scarlet fever, and invasive diseases. This study analyzed the incidence and characteristics of S. pyogenes infections between 2018 and 2023, examining 915 cases categorized as either respiratory or non-respiratory. Respiratory infections predominantly affected children, accounting for 76% of cases, with a median age of 5 [3, 8] years, while non-respiratory infections were more common in adults, with a median age of 46.5 [34, 64] years. Invasive respiratory infections, such as pneumonia and empyema, were more frequent in children (54.8%), whereas invasive non-respiratory infections, such as primarily cellulitis, were predominantly seen in adults (90.5%). A sharp decline in S. pyogenes infections was observed during the pandemic, with respiratory cases decreasing tenfold in 2020 compared to the previous year, and non-respiratory cases experiencing a twofold reduction. However, infection rates returned to pre-pandemic levels by 2022 and 2023, with a notable resurgence of invasive respiratory infections in children following a public health alert in the United Kingdom in late 2022. These findings highlight distinct infection patterns between pediatric and adult populations and emphasize the significant impact of the pandemic on respiratory infections, particularly in children.

Abstract Surveillance data published since 2010, although limited, showed that there is no evidence of zoonotic parasite infection in market quality Atlantic salmon, marine rainbow trout, gilthead seabream, turbot, meagre, Atlantic halibut, common carp and European catfish. No studies were found for greater amberjack, brown trout, African catfish, European eel and pikeperch. Anisakis pegreffii, A. simplex (s. s.) and Cryptocotyle lingua were found in European seabass, Atlantic bluefin tuna and/or cod, and Pseudamphistomum truncatum and Paracoenogonimus ovatus in tench, produced in open offshore cages or flow‐through ponds or tanks. It is almost certain that fish produced in closed recirculating aquaculture systems (RAS) or flow‐through facilities with filtered water intake and exclusively fed heat‐treated feed are free of zoonotic parasites. Since the last EFSA opinion, the UV‐press and artificial digestion methods have been developed into ISO standards to detect parasites in fish, while new UV‐scanning, optical, molecular and OMICs technologies and methodologies have been developed for the detection, visualisation, isolation and/or identification of zoonotic parasites in fish. Freezing and heating continue to be the most efficient methods to kill parasites in fishery products. High‐pressure processing may be suitable for some specific products. Pulsed electric field is a promising technology although further development is needed. Ultrasound treatments were not effective. Traditional dry salting of anchovies successfully inactivated Anisakis. Studies on other traditional processes – air‐drying and double salting (brine salting plus dry salting) – suggest that anisakids are successfully inactivated, but more data covering these and other parasites in more fish species and products is required to determine if these processes are always effective. Marinade combinations with anchovies have not effectively inactivated anisakids. Natural products, essential oils and plant extracts, may kill parasites but safety and organoleptic data are lacking. Advanced processing techniques for intelligent gutting and trimming are being developed to remove parasites from fish.

Abstract The utility of reverse transcription-polymerase chain reaction (RT-PCR) in analysis SARS-COV-2 variants was evaluated. RT-PCR tests were used to analyse the majority of new SARS-CoV-2 cases (n = 9315) in a tertiary hospital (Madrid, Spain) throughout 2021. Subsequently, whole genome sequencing (WGS) was conducted on 10.8% of these samples (n = 1002). Notably, the Delta and Omicron variants emerged rapidly. There were no discrepancies between RT-PCR and WGS results. Continuous surveillance of SARS-CoV-2 variants is essential, and RT-PCR is a highly useful method, specially during periods of high COVID-19 incidence. This feasible technique can be implemented in all SARS-CoV-2 laboratories. However, WGS remains the gold standard method for comprehensive detection of all existing SARS-CoV-2 variants.

Summary: Background: Anti-COVID-19 hyperimmune immunoglobulin (hIG) can provide standardized and controlled antibody content. Data from controlled clinical trials using hIG for the prevention or treatment of COVID-19 outpatients have not been reported. We assessed the safety and efficacy of subcutaneous anti-COVID-19 hyperimmune immunoglobulin 20% (C19-IG20%) compared to placebo in preventing development of symptomatic COVID-19 in asymptomatic individuals with SARS-CoV-2 infection. Methods: We did a multicentre, randomized, double-blind, placebo-controlled trial, in asymptomatic unvaccinated adults (≥18 years of age) with confirmed SARS-CoV-2 infection within 5 days between April 28 and December 27, 2021. Participants were randomly assigned (1:1:1) to receive a blinded subcutaneous infusion of 10 mL with 1 g or 2 g of C19-IG20%, or an equivalent volume of saline as placebo. The primary endpoint was the proportion of participants who remained asymptomatic through day 14 after infusion. Secondary endpoints included the proportion of individuals who required oxygen supplementation, any medically attended visit, hospitalisation, or ICU, and viral load reduction and viral clearance in nasopharyngeal swabs. Safety was assessed as the proportion of patients with adverse events. The trial was terminated early due to a lack of potential benefit in the target population in a planned interim analysis conducted in December 2021. ClinicalTrials.gov registry: NCT04847141. Findings: 461 individuals (mean age 39.6 years [SD 12.8]) were randomized and received the intervention within a mean of 3.1 (SD 1.27) days from a positive SARS-CoV-2 test. In the prespecified modified intention-to-treat analysis that included only participants who received a subcutaneous infusion, the primary outcome occurred in 59.9% (91/152) of participants receiving 1 g C19-IG20%, 64.7% (99/153) receiving 2 g, and 63.5% (99/156) receiving placebo (difference in proportions 1 g C19-IG20% vs. placebo, −3.6%; 95% CI -14.6% to 7.3%, p = 0.53; 2 g C19-IG20% vs placebo, 1.1%; −9.6% to 11.9%, p = 0.85). None of the secondary clinical efficacy endpoints or virological endpoints were significantly different between study groups. Adverse event rate was similar between groups, and no severe or life-threatening adverse events related to investigational product infusion were reported. Interpretation: Our findings suggested that administration of subcutaneous human hyperimmune immunoglobulin C19-IG20% to asymptomatic individuals with SARS-CoV-2 infection was safe but did not prevent development of symptomatic COVID-19. Funding: Grifols.

Abstract Between October 2021 and September 2022 Europe has suffered the most devastating highly pathogenic avian influenza (HPAI) epidemic with a total of 2,520 outbreaks in poultry, 227 outbreaks in captive birds, and 3,867 HPAI virus detections in wild birds. The unprecedent geographical extent (37 European countries affected) resulted in 50 million birds culled in affected establishments. In the current reporting period, between 10 September and 2 December 2022, 1,163 HPAI virus detections were reported in 27 European countries in poultry (398), captive (151) and wild birds (613). A decrease in HPAI virus detections in colony‐breeding seabirds species and an increase in the number of detections in waterfowl has been observed. The continuous circulation of the virus in the wild reservoir has led to the frequent introduction of the virus into poultry populations. It is suspected that waterfowl might be more involved than seabirds in the incursion of HPAI virus into poultry establishments. In the coming months, the increasing infection pressure on poultry establishments might increase the risk of incursions in poultry, with potential further spread, primarily in areas with high poultry densities. The viruses detected since September 2022 (clade 2.3.4.4b) belong to eleven genotypes, three of which have circulated in Europe during the summer months, while eight represent new genotypes. HPAI viruses were also detected in wild and farmed mammal species in Europe and North America, showing genetic markers of adaptation to replication in mammals. Since the last report, two A(H5N1) detections in humans in Spain, one A(H5N1), one A(H5N6) and one A(H9N2) human infection in China as well as one A(H5) infection without NA‐type result in Vietnam were reported, respectively. The risk of infection is assessed as low for the general population in the EU/EEA, and low to medium for occupationally exposed people.

Here we focus on designing, for the first time, microsatellite markers for evolutionary and ecological research on aquatic beetles from the genus Ochthebius (Coleoptera, Hydraenidae). Some of these non-model species, with high cryptic diversity, exclusively inhabit supratidal rockpools, extreme and highly dynamic habitats with important anthropogenic threats. We analysed 15 individuals of four species (O. lejolisii, O. subinteger, O. celatus, and O. quadricollis) across 10 localities from the Mediterranean coasts of Spain and Malta. Using next-generation sequencing technology, two libraries were constructed to interpret the species of the two subgenera present consistently (Ochthebius s. str., O. quadricollis; and Cobalius, the rest of the species). Finally, 20 markers (10 for each subgenus) were obtained and successfully tested by cross-validation in the four species under study. As a by-catch, we could retrieve the complete mitochondrial genomes of O. lejolisii, O. quadricollis, and O. subinteger. Interestingly, the mitochondrial genome of O. quadricollis exhibited high genetic variability compared to already published data. The novel SSR panels and mitochondrial genomes for Ochthebius will be valuable in future research on species identification, diversity, genetic structure, and population connectivity in highly dynamic and threatened habitats such as supratidal coastal rockpools.

Abstract The bumblebee Bombus terrestris is used worldwide for crop pollination. Despite its positive impact on crop yield, it has become a widespread threat to biodiversity due to its interactions with local bumblebee populations. Commercial subspecies introduced to the Iberian Peninsula since the 1990s without any regulation have colonized the environment, with evidence of naturalization and introgression with the endemic subspecies Bombus terrestris lusitanicus. We have used mitochondrial and nuclear genetic data to describe the current genetic diversity of the Iberian population and to estimate the expansion of commercial bumblebees. Samples from the natural distribution range of the commercial subspecies, the natural intergradation area between the two subspecies and from a period prior to the use of commercial colonies (i.e., before the 1990s) have been used for comparison. Our results show that the mitochondrial haplotype of the commercial breeds has spread throughout the territory, which, together with subtle changes observed in the nuclear genetic diversity of the populations, indicates that hybridization and consequent introgression are occurring in most of the peninsula. It is, therefore, necessary to improve the existing legislation concerning the management and exportation of commercial bumblebees to conserve locally adapted populations.

Abstract The 2021–2022 highly pathogenic avian influenza (HPAI) epidemic season is the largest HPAI epidemic so far observed in Europe, with a total of 2,467 outbreaks in poultry, 47.7 million birds culled in the affected establishments, 187 outbreaks in captive birds, and 3,573 HPAI virus detections in wild birds with an unprecedent geographical extent reaching from Svalbard islands to South Portugal and Ukraine, affecting 37 European countries. Between 11 June and 9 September 2022, 788 HPAI virus detections were reported in 16 European countries in poultry (56), captive (22) and wild birds (710). Several colony‐breeding seabird species exhibited widespread and massive mortality from HPAI A(H5N1) virus along the northwest coast of Europe. This resulted in an unprecedentedly high level of HPAI virus detections in wild birds between June and August 2022 and represents an ongoing risk of infection for domestic birds. HPAI outbreaks were still observed in poultry from June to September with five‐fold more infected premises than observed during the same period in 2021 and mostly distributed along the Atlantic coast. Response options to this new epidemiological situation include the definition and rapid implementation of suitable and sustainable HPAI mitigation strategies such as appropriate biosecurity measures and surveillance strategies for early detection in the different poultry production systems. The viruses currently circulating in Europe belong to clade 2.3.4.4b with seven genotypes, three of which identified for the first time during this time period, being detected during summer. HPAI A(H5) viruses were also detected in wild mammal species in Europe and North America and showed genetic markers of adaptation to replication in mammals. Since the last report, two A(H5N6), two A(H9N2) and one A(H10N3) human infections were reported in China. The risk of infection is assessed as low for the general population in the EU/EEA, and low to medium for occupationally exposed people.

Abstract The 2021–2022 highly pathogenic avian influenza (HPAI) epidemic season is the largest epidemic so far observed in Europe, with a total of 2,398 outbreaks in poultry, 46 million birds culled in the affected establishments, 168 detections in captive birds, and 2,733 HPAI events in wild birds in 36 European countries. Between 16 March and 10 June 2022, 1,182 HPAI virus detections were reported in 28 EU/EEA countries and United Kingdom in poultry (750), and in wild (410) and captive birds (22). During this reporting period, 86% of the poultry outbreaks were secondary due to between‐farm spread of HPAI virus. France accounted for 68% of the overall poultry outbreaks, Hungary for 24% and all other affected countries for less than 2% each. Most detections in wild birds were reported by Germany (158), followed by the Netherlands (98) and the United Kingdom (48). The observed persistence of HPAI (H5) virus in wild birds since the 2020–2021 epidemic wave indicates that it may have become endemic in wild bird populations in Europe, implying that the health risk from HPAI A(H5) for poultry, humans, and wildlife in Europe remains present year‐round, with the highest risk in the autumn and winter months. Response options to this new epidemiological situation include the definition and the rapid implementation of suitable and sustainable HPAI mitigation strategies such as appropriate biosecurity measures and surveillance strategies for early detection measures in the different poultry production systems. Medium to long‐term strategies for reducing poultry density in high‐risk areas should also be considered. The results of the genetic analysis indicate that the viruses currently circulating in Europe belong to clade 2.3.4.4b. HPAI A(H5) viruses were also detected in wild mammal species in Canada, USA and Japan, and showed genetic markers of adaptation to replication in mammals. Since the last report, four A(H5N6), two A(H9N2) and two A(H3N8) human infections were reported in China and one A(H5N1) in USA. The risk of infection is assessed as low for the general population in the EU/EEA, and low to medium for occupationally exposed people.

Frente a la pandemia actual ocasionada por el coronavirus se crea una situación de crisis humanitaria, haciéndose imprescindible considerar los aspectos éticos en el otorgamiento de Cuidados Paliativos para brindar atención integral a la población al final de la vida. Entendiendo la Enfermería como un pilar en el equipo de trabajo de los Cuidados Paliativos, el presente ensayo tiene como objetivo reflexionar sobre la Ética en Enfermería en Cuidados Paliativos al final de la vida en tiempos de COVID-19. Los Cuidados Paliativos se desarrollan bajo los principios éticos, entendiendo que todas las personas deben tener acceso universal a salud, incluso cuando la enfermedad se encuentra en etapa avanzada; brindando calidad de vida y acompañamiento en una muerte digna. Sin embargo, frente al alto grado de contagio del coronavirus, personas mueren aisladas en servicios de cuidados intensivos sin el acompañamiento de la familia; siendo responsabilidad del equipo de salud otorgar compañía en la muerte. Ante esta realidad, son necesarias actividades de autocuidado para el equipo de salud y la familia, además, del aseguramiento de las condiciones de seguridad en el trabajo y el uso de equipo de protección. Es así como Enfermería tiene un rol ético fundamental en la crisis humanitaria actual, principalmente la entrega de Cuidados Paliativos al final de la vida en tiempos de COVID-19, acompañando en el final de la vida, evitando llegar al aislamiento e incluso la muerte en soledad.

Abstract Teosinte, wild maize relatives originating from Mexico and Central America, emerged as a noxious agricultural weed in France and Spain. In 2016, the European Food Safety Authority (EFSA) issued a technical report that assessed the available scientific information on teosinte for its relevance for the environmental risk assessment (ERA) and risk management (RM) of genetically modified (GM) maize MON810, Bt11, 1507 and GA21 for cultivation. It was concluded that the impact of insect resistance and/or herbicide tolerance in GM teosinte hybrid progeny (potentially acquired through hybridisation between GM maize and teosinte) on target and non‐target organisms, the abiotic environment and biogeochemical cycles would be very low under EU conditions. Following a request of the European Commission, EFSA evaluated whether the ERA conclusions and RM recommendations of EFSA (2016) remain applicable, or require revision in light of new scientific evidence on teosinte that has become available since the publication of EFSA (2016). A protocol was developed to clarify the interpretation of the terms of reference of the mandate and make them operational. The assessment relied on evidence retrieved via an extensive literature search and from reports of the Competent Authorities of France and Spain, and on hearing expert testimonies. A limited collection of 18 publications of varying relevance and quality was retrieved and assessed. Based on this evidence, it is concluded that the ERA conclusions and RM recommendations of EFSA (2016) remain applicable, except those pertaining to the use of glyphosate‐based herbicides on maize GA21 which should be considered under Regulation (EC) No 1107/2009. In infested agricultural areas (especially in regions where maize MON810 is widely grown), weed management measures implemented to monitor, control and/or eradicate teosinte must remain in place, as they will contribute to further reduce the low vertical gene flow potential between GM maize and EU teosinte.

Abstract Between 9 December 2021 and 15 March 2022, 2,653 highly pathogenic avian influenza (HPAI) virus detections were reported in 33 EU/EEA countries and the UK in poultry (1,030), in wild (1,489) and in captive birds (133). The outbreaks in poultry were mainly reported by France (609), where two spatiotemporal clusters have been identified since October 2021, followed by Italy (131), Hungary (73) and Poland (53); those reporting countries accounted together for 12.8 of the 17.5 million birds that were culled in the HPAI affected poultry establishments in this reporting period. The majority of the detections in wild birds were reported by Germany (767), the Netherlands (293), the UK (118) and Denmark (74). HPAI A(H5) was detected in a wide range of host species in wild birds, indicating an increasing and changing risk for virus incursion into poultry farms. The observed persistence and continuous circulation of HPAI viruses in migratory and resident wild birds will continue to pose a risk for the poultry industry in Europe for the coming months. This requires the definition and the rapid implementation of suitable and sustainable HPAI mitigation strategies such as appropriate biosecurity measures, surveillance plans and early detection measures in the different poultry production systems. The results of the genetic analysis indicate that the viruses currently circulating in Europe belong to clade 2.3.4.4b. Some of these viruses were also detected in wild mammal species in the Netherlands, Slovenia, Finland and Ireland showing genetic markers of adaptation to replication in mammals. Since the last report, the UK reported one human infection with A(H5N1), China 17 human infections with A(H5N6), and China and Cambodia 15 infections with A(H9N2) virus. The risk of infection for the general population in the EU/EEA is assessed as low, and for occupationally exposed people, low to medium.

Abstract Following a request from five European Nordic countries, the EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA) was tasked to provide scientific advice on a tolerable upper intake level (UL) or a safe level of intake for dietary (total/added/free) sugars based on available data on chronic metabolic diseases, pregnancy‐related endpoints and dental caries. Specific sugar types (fructose) and sources of sugars were also addressed. The intake of dietary sugars is a well‐established hazard in relation to dental caries in humans. Based on a systematic review of the literature, prospective cohort studies do not support a positive relationship between the intake of dietary sugars, in isocaloric exchange with other macronutrients, and any of the chronic metabolic diseases or pregnancy‐related endpoints assessed. Based on randomised control trials on surrogate disease endpoints, there is evidence for a positive and causal relationship between the intake of added/free sugars and risk of some chronic metabolic diseases: The level of certainty is moderate for obesity and dyslipidaemia (> 50–75% probability), low for non‐alcoholic fatty liver disease and type 2 diabetes (> 15–50% probability) and very low for hypertension (0–15% probability). Health effects of added vs. free sugars could not be compared. A level of sugars intake at which the risk of dental caries/chronic metabolic diseases is not increased could not be identified over the range of observed intakes, and thus, a UL or a safe level of intake could not be set. Based on available data and related uncertainties, the intake of added and free sugars should be as low as possible in the context of a nutritionally adequate diet. Decreasing the intake of added and free sugars would decrease the intake of total sugars to a similar extent. This opinion can assist EU Member States in setting national goals/recommendations.

Abstract The 2020–2021 avian influenza epidemic with a total of 3,777 reported highly pathogenic avian influenza (HPAI) detections and approximately 22,900,000 affected poultry birds in 31 European Countries appears to be one of the largest HPAI epidemics that has ever occurred in Europe. Between 15 May and 15 September 2021, 162 HPAI virus detections were reported in 17 EU/EEA countries and the UK in poultry (51), in wild (91) and captive birds (20). The detections in poultry were mainly reported by Kosovo (20), Poland (17) and Albania (6). HPAI virus was detected during the summer months in resident wild bird populations mainly in northern Europe. The data presented in this report indicates that HPAI virus is still circulating in domestic and wild bird populations in some European countries and that the epidemic is not over yet. Based on these observations, it appears that the persistence of HPAI A(H5) in Europe continues to pose a risk of further virus incursions in domestic bird populations. Furthermore, during summer, HPAI viruses were detected in poultry and several wild bird species in areas in Russia that are linked to key migration areas of wild waterbirds; this is of concern due to the possible introduction and spread of novel virus strains via wild birds migrating to the EU countries during the autumn from the eastern breeding to the overwintering sites. Nineteen different virus genotypes have been identified so far in Europe and Central Asia since July 2020, confirming a high propensity for this virus to undergo reassortment events. Since the last report, 15 human infections due to A(H5N6) HPAI and five human cases due to A(H9N2) low pathogenic avian influenza (LPAI) virus have been reported from China. Some of these cases were caused by a virus with an HA gene closely related to the A(H5) viruses circulating in Europe. The viruses characterised to date retain a preference for avian‐type receptors; however, the reports of transmission events of A(H5) viruses to mammals and humans in Russia, as well as the recent A(H5N6) human cases in China may indicate a continuous risk of these viruses adapting to mammals. The risk of infection for the general population in the EU/EEA is assessed as very low, and for occupationally exposed people low, with large uncertainty due to the high diversity of circulating viruses in the bird populations.

Abstract The 2020‐2021 epidemic with a total of 3,555 reported HPAI detections and around 22,400,000 affected poultry birds in 28 European Countries appears to be one of the largest and most devastating HPAI epidemics ever occurred in Europe. Between 24 February and 14 May 2021, 1,672 highly pathogenic avian influenza (HPAI) virus detections were reported in 24 EU/EEA countries and the UK in poultry (n=580), and in wild (n=1,051) and captive birds (n=41). The majority of the detections in poultry were reported by Poland that accounted for 297 outbreaks occurring in a densely populated poultry area over a short period of time, followed by Germany with 168 outbreaks. Germany accounted for 603 detections in wild birds, followed by Denmark and Poland with 167 and 56 detections, respectively. A second peak of HPAI‐associated wild bird mortality was observed from February to April 2021 in north‐west Europe. The observed longer persistence of HPAI in wild birds compared to previous years may result in a continuation of the risk for juveniles of wild birds and mammals, as well as for virus entry into poultry farms. Therefore, enhanced awareness among farmers to continue applying stringent biosecurity measures and to monitor and report increases in daily mortality and drops in production parameters, are recommended. Sixteen different genotypes were identified to date in Europe and Russia, suggesting a high propensity of these viruses to reassort. The viruses characterized to date retain a preference for avian‐type receptors; however, transmission events to mammals and the identification of sporadic mutations of mammal adaptation, indicate ongoing evolution processes and possible increased ability of viruses within this clade to further adapt and transmit to mammals including humans. Since the last report, two human infections due to A(H5N6) HPAI were reported from China and Laos and 10 human cases due to A(H9N2) low pathogenic avian influenza (LPAI) virus identified in China and Cambodia. The risk of infection for the general population in the EU/EEA is assessed as very low and for occupationally exposed people low. People exposed during avian influenza outbreaks should adhere to protection measures, strictly wear personal protective equipment and get tested immediately when developing respiratory symptoms or conjunctivitis within 10 days after exposure.

Abstract The European Commission requested EFSA to assess if different thermal processes achieve a 5 log10 reduction in Enterococcus faecalis or Salmonella Senftenberg (775W) and (if relevant) a 3 log10 reduction in thermoresistant viruses (e.g. Parvovirus) as well as if different chemical processes achieve a 3 log10 reduction of eggs of Ascaris sp., in eight groups of Category 2 and 3 derived products and animal by‐products (ABP). These included (1) ash derived from incineration, co‐incineration and combustion; (2) glycerine derived from the production of biodiesel and renewable fuels; (3) other materials derived from the production of biodiesel and renewable fuels; (4) hides and skins; (5) wool and hair; (6) feathers and down; (7) pig bristles; and (8) horns, horn products, hooves and hoof products. Data on the presence of viral hazards and on thermal and chemical inactivation of the targeted indicator microorganisms and biological hazards under relevant processing conditions were extracted via extensive literature searches. The evidence was assessed via expert knowledge elicitation. The certainty that the required log10 reductions in the most resistant indicator microorganisms or biological hazards will be achieved for each of the eight groups of materials mentioned above by the thermal and/or chemical processes was (1) 99–100% for the two processes assessed; (2) 98–100% in Category 2 ABP, at least 90–99% in Category 3 ABP; (3) 90–99% in Category 2 ABP; at least 66–90% in Category 3 ABP; (4) 10–66% and 33–66%; (5) 1–33% and 10–50%; (6) 66–90%; (7) 33–66% and 50–95%; (8) 66–95%, respectively. Data generation on the occurrence and reduction of biological hazards by thermal and/or chemical methods in these materials and on the characterisation of the usage pathways of ABP as organic fertilisers/soil improvers is recommended.

Abstract Between 16 September and 8 December 2021, 867 highly pathogenic avian influenza (HPAI) virus detections were reported in 27 EU/EEA countries and the UK in poultry (316), in wild (523) and in captive birds (28). The detections in poultry were mainly reported by Italy (167) followed by Hungary and Poland (35 each). Tha majority of the detections in wild birds were reported by Germany (280), Netherlands (65) and United Kingdom (53). The observed persistence and continuous circulation of HPAI viruses in migratory and resident wild birds will continue to pose a risk for the poultry industry in Europe for the coming months. The frequent occurrence of HPAI A(H5) incursions in commercial farms (including poultry production types considered at low avian influenza risk) raises concern about the capacity of the applied biosecurity measures to prevent virus introduction. Short‐term preparedness and medium‐ and long‐term prevention strategies, including revising and reinforcing biosecurity measures, reduction of the density of commercial poultry farms and possible appropriate vaccination strategies, should be implemented. The results of the genetic analysis indicate that the viruses characterised during this reporting period belong to clade 2.3.4.4b. Some of the characterized HPAI A(H5N1) viruses detected in Sweden, Germany, Poland and United Kingdom are related to the viruses which have been circulating in Europe since October 2020; in North, Central, South and East Europe novel reassortant A(H5N1) virus has been introduced starting from October 2021. HPAI A(H5N1) was also detected in wild mammal species in Sweden, Estonia and Finland; some of these strains characterised so far present an adaptive marker that is associated with increased virulence and replication in mammals. Since the last report, 13 human infections due to HPAI A(H5N6) and two human cases due to LPAI A(H9N2) virus have been reported from China. Some of these A(H5N6) cases were caused by a reassortant virus of clade 2.3.4.4b, which possessed an HA gene closely related to the A(H5) viruses circulating in Europe. The risk of infection for the general population in the EU/EEA is assessed as low, and for occupationally exposed people, low to medium, with large uncertainty due to the high diversity of circulating viruses in the bird populations.

The unique composition and technological properties of some oat bran components (mainly protein and soluble fiber) and olive oil make them a good choice to form oil-in-water vegetable emulsions. The different concentrations of oat bran were studied to form olive oil-in water (O/W) emulsions to apply as a replacement for fat and meat. As a result, four O/W emulsions (OBE) were formulated with 10% (OBE10), 15% (OBE15), 20% (OEB20), and 30% (OBE30) oat bran concentrations and 40% olive oil, with the corresponding amount of water added for each O/W emulsion. Composition, technological properties (thermal stability, pH, texture), and lipid structural characteristics were evaluated. The results showed that low oat bran content (OEB10)—with a lower concentration of oat protein and β-glucans—resulted in an O/W emulsion with an aggregated droplet structure and lower thermal stability and hardness. These connections between composition, technology, and structural properties of olive O/W emulsions elaborated with oat bran could help in making the optimal choice for their potential application in the production of foods such as healthier meat products.

Background: Decidualization of the uterine mucosa drives the maternal adaptation to invasion by the placenta. Appropriate depth of placental invasion is needed to support a healthy pregnancy; shallow invasion is associated with the development of severe preeclampsia (sPE). Maternal contribution to sPE through failed decidualization is an important determinant of placental phenotype. However, the molecular mechanism underlying the in vivo defect linking decidualization to sPE is unknown. Methods: Global RNA sequencing was applied to obtain the transcriptomic profile of endometrial biopsies collected from nonpregnant women who suffer sPE in a previous pregnancy and women who did not develop this condition. Samples were randomized in two cohorts, the training and the test set, to identify the fingerprinting encoding defective decidualization in sPE and its subsequent validation. Gene Ontology enrichment and an interaction network were performed to deepen in pathways impaired by genetic dysregulation in sPE. Finally, the main modulators of decidualization, estrogen receptor 1 (ESR1) and progesterone receptor B (PGR-B), were assessed at the level of gene expression and protein abundance. Results: Here, we discover the footprint encoding this decidualization defect comprising 120 genes—using global gene expression profiling in decidua from women who developed sPE in a previous pregnancy. This signature allowed us to effectively segregate samples into sPE and control groups. ESR1 and PGR were highly interconnected with the dynamic network of the defective decidualization fingerprint. ESR1 and PGR-B gene expression and protein abundance were remarkably disrupted in sPE. Conclusions: Thus, the transcriptomic signature of impaired decidualization implicates dysregulated hormonal signaling in the decidual endometria in women who developed sPE. These findings reveal a potential footprint that could be leveraged for a preconception or early prenatal screening of sPE risk, thus improving prevention and early treatments. Funding: This work has been supported by the grant PI19/01659 (MCIU/AEI/FEDER, UE) from the Spanish Carlos III Institute awarded to TGG. NCM was supported by the PhD program FDGENT/2019/008 from the Spanish Generalitat Valenciana. IMB was supported by the PhD program PRE2019-090770 and funding was provided by the grant RTI2018-094946-B-100 (MCIU/AEI/FEDER, UE) from the Spanish Ministry of Science and Innovation with CS as principal investigator. This research was funded partially by Igenomix S.L.

Abstract Background Ladybird beetles (Coleoptera: Coccinellidae) are abundant predatory species in many agroecosystems, are valued for their biological pest control functions, and have been recommended as test species for studies supporting the assessment of non-target effects of insect-resistant Bt crops. Although insecticidal Bt proteins are known to be highly specific against target pests, some recent laboratory studies reported putative toxic effects of Bt proteins on ladybird species. While such studies have been criticised because of methodological shortcomings or inconsistencies, they cast doubt on the insecticidal spectrum of activity of some Bt proteins. Performing a systematic review that synthesises all existing evidence on this controversial topic may help to resolve the remaining scientific uncertainties. The review question to be addressed by the systematic review is the following: Are ladybird beetles (Coleoptera: Coccinellidae) affected by Bt proteins expressed in genetically modified insect-resistant crops? The systematic review will focus on studies performed under controlled environmental conditions. Methods An extensive literature search will be conducted to identify the articles relevant to the review question. A wide range of electronic bibliographic databases, the internet search engine Google Scholar, and websites of specialized organizations will be searched. Citation searching, reference list-checking and searching of key journals will also be performed. The relevance of the identified articles will be assessed against a set of pre-defined eligibility criteria, following a two-step approach. In the first step, title and abstract (or summary) will be screened, whilst in the second step the full text of all remaining articles will be assessed by two members of the review team. All relevant studies will be subjected to an appraisal of external (generalisability) and internal (risk of bias) validity. Data from the selected studies will be extracted and synthesised in a narrative report. If a sufficient number of datasets generated with comparable experimental setup is available, statistical meta-analyses will be conducted on a range of comparisons and including sensitivity analyses.

One of the main goals of neuromorphic computing is the implementation and design of systems capable of dynamic evolution with respect to their own experience. In biology, synaptic scaling is the homeostatic mechanism which controls the frequency of neural spikes within stable boundaries for improved learning activity. To introduce such control mechanism in a hardware spiking neural network (SNN), we present here a novel artificial neuron based on phase change memory (PCM) devices capable of internal regulation via homeostatic and plastic phenomena. We experimentally show that this mechanism increases the robustness of the system thus optimizing the multi-pattern learning under spike-timing-dependent plasticity (STDP). It also improves the continual learning capability of hybrid supervised-unsupervised convolutional neural networks (CNNs), in terms of both resilience and accuracy. Furthermore, the use of neurons capable of self-regulating their fire responsivity as a function of the PCM internal state enables the design of dynamic networks. In this scenario, we propose to use the PCM-based neurons to design bio-inspired recurrent networks for autonomous decision making in navigation tasks. The agent relies on neuronal spike-frequency adaptation (SFA) to explore the environment via penalties and rewards. Finally, we show that the conductance drift of the PCM devices, contrarily to the applications in neural network accelerators, can improve the overall energy efficiency of neuromorphic computing by implementing bio-plausible active forgetting.

The taxonomic status of Bombus terrestris subspecies is complex and has deep implications in the management of commercial bumblebees for crop pollination as well as in the establishment of appropriate conservation plans. Herein, the complete mitogenome of the endemic Canary Islands subspecies Bombus terrestris canariensis is newly sequenced and compared with available mitochondrial sequences in order to shed light into its taxonomic status. The mitochondrial genome was 17,300 bp in length and contained 37 genes, including 13 protein-coding genes (PCGs), two rRNAs, and 22 tRNAs and a partial sequence of the AT rich control region. The phylogenetic analysis of PCGs of the mitogenome was congruent with its subspecific status and a close relationship with the North African subspecies africanus as previously suggested. The sequencing of the mitogenome of B. t. canariensis provides useful genetic information to study the conservation genetics and genetic diversity of these island bumblebee populations.

Abstract The European Commission requested EFSA assess antimicrobial‐resistant bacteria responsible for animal transmissible diseases, with a view to listing such pathogens for European Union action. This Scientific Opinion addresses the ad hoc method developed: (i) to give a global state of play as regards resistant animal pathogens that cause transmissible animal diseases, (ii) to identify the most relevant bacteria in the EU and (iii) to summarise their actual or potential animal health impact, and to perform their assessment for being listed and categorised according to the criteria of Articles 7, 5, 9 and 8 within the Animal Health Law (AHL) framework. An extensive literature review is carried out to give the global state of play of selected resistant bacteria that constitute a threat to animal health (i). An expert judgement procedure, based on the outcome of the literature review, is applied to identify which among those bacteria subjected to the literature review are the ‘most relevant’ in the European Union (ii). Their animal health impact in the European Union and their assessment for being listed and categorised according to the AHL framework will follow the ‘ad hoc method for the assessment on listing and categorisation of animal diseases within the framework of the Animal Health Law’ that EFSA has developed in the past (iii). The assessment of (i) and (ii) is addressed in distinct scientific opinions that are published separately by animal species of interest (dogs and cats, horses, pigs, poultry, cattle, small ruminants, rabbits and aquatic animal species). The assessment of (iii) is addressed in distinct scientific opinions and published separately by the animal pathogen.

Between 8 December 2020 and 23 February 2021, 1,022 highly pathogenic avian influenza (HPAI) virus detectionswere reported in 25 EU/EEA countries and the UK in poultry (n=592), wild (n=421) and captive birds (n=9).The majority of the detections were reported by Francethat accounted for 442 outbreaks in poultry,mostly located inthe Landes regionandaffecting the foie gras production industry,and six wild bird detections; Germany,who reported 207 detections in wild birds and 50 poultry outbreaks; Denmark,with 63 detections in wild birds and one poultry outbreak; and Poland,with 37 poultry outbreaks and 24 wild bird detections. Due to the continued presence of HPAI A(H5) viruses in wild birds and the environment,there is still a risk of avian influenza incursions with the potential further spread between establishments, primarily in areas with high poultry densities. As the currently circulating HPAI A(H5N8) virus cancause high mortality also in affected duck farms, mortality eventscan be seen as a good indicator of virus presence. However,also subclinical virusspread in this type of poultry production system have been reported.To improve early detection of infection in poultry within the surveillance zone, the clinical inspection of duck establishments should be complemented by encouraging farmers to collect dead birds to be pooled and tested weekly (bucket sampling).Six different genotypes were identified to date in Europe and Russia, suggesting a high propensity of these viruses to undergo multiple reassortment events. To date, no evidence of fixation of known mutations previously described as associated to zoonotic potential has been observed in HPAI viruses currently circulanting in Europe based on the available sequences.Seven cases due to A(H5N8) HPAI virus have been reported from Russia, all were poultry workerswith mild or no symptoms. Five human cases due to A(H5N6) HPAI and 10 cases due to A(H9N2) LPAI viruseshave been reported from China. The risk for the general population as well as travel‐related imported human cases is assessed as very lowand the risk forpeople occupationally exposedpeople as low.Any human infections with avian influenza viruses are notifiablewithin 24 hoursthrough the Early Warning and Response System (EWRS) and the International Health Regulations (IHR) notification system.

Background: Persistent and relapsing prosthetic joint infection (PJI) due to Staphylococcus aureus presents a clinical challenge. This study aimed to provide an extensive description of phenotypic and genomic changes that could be related to persistence or relapse. Methods: Initial and second S. aureus isolates from 6 cases of persistent and relapsing PJI, along with clinical isolates from 8 cases, with favorable outcome were included. All isolates were studied by phenotypic and genotypic approaches. Results: Recurrent S. aureus isolates exhibited a significant increase in adhesive capacity, invasion and persistence compared to resolved isolates. No association was found for the presence or absence of certain genes with the persistence or relapse of PJI. All sequential isolates showed identical sequence type (ST). Resistance gene loss during the infection and a great diversity of variants in different virulence genes between the pair of strains, mainly in genes encoding adhesins such as fnbA, were observed. Conclusions: S. aureus-caused relapse and persistence PJI is associated with bacterial phenotypical and genotypical adaptation. The main paths of adaptation were persistence in the intracellular compartment, and the loss of antibiotic resistance genes and variant acquisition, especially in genes encoding adhesins.

Abstract This guidance document is intended to assist the applicant in the preparation and the presentation of an application, as foreseen in Article 7.6 of Regulation (EC) No 1831/2003, for the renewal of the authorisation of additives for use in animal nutrition. Draft Endorsed by the FEEDAP Panel 14 November 2019 Submitted for public consultation 29 November 2019 End of public consultation 26 January 2020 Adoption by the FEEDAP Panel 19 November 2020 Implementation date 27 March 2021

Between 16 May and 15 August 2020, seven highly pathogenic avian influenza (HPAI) A(H5N8) virus outbreaks were reported in Europe in poultry, with one outbreak reported in Bulgaria(n=1) andsix in Hungary (n=6) and one low pathogenic avian influenza (LPAI) A(H5N3) virus outbreak was reported in poultry in Italy. All six outbreaks detected in Hungary were secondary outbreaks and seem to be the tail end of the HPAI A(H5N8) epidemic that wasobserved in poultry over the winter and spring in central Europe from December 2019 (n=334).Genetic analysis of the HPAI A(H5N8) viruses isolated during this reporting period from Bulgaria and Hungary did not identify any major changes compared tothe viruses collected in the respective countries during the first months of 2020. This suggests a persistence of the virus in the two countries rather than new introductions via infectedwild birds. HPAI A(H5N8) virus has been detected in poultry and wild birds in western Russia within the reporting period, and as of the middle of September also in Kazakhstan. The presence of HPAI virus in western Russiaand in north Kazakhstan,spatially associated with autumnmigration routes of wild waterbirds, is of concern due to the possible spread of the virus via wild birds migrating to the EU.It is highly recommended thatMember States take appropriate measures to promptly detect suspected cases of HPAI, including increasing biosecurity measures. According to past experiences (2005‐2006 and 2016‐2017 epidemic waves), the northern and eastern European areas might be at higher risk of virus introduction in the coming autumn‐winter seasonand should be the key regions where prompt response measures to early detect the virusshould be set up. One human case due to A(H9N2) avian influenza virus infection was reported during the reporting period.

Between 16 February and 15 May 2020, 290highly pathogenic avian influenza (HPAI) A(H5) virus outbreakswere reported in Europe in poultry (n=287), captive birds (n=2) and wild birds (n=1)in Bulgaria, Czechia,Germany,Hungary andPolandand two low pathogenic avian influenza (LPAI) A(H7N1) virus outbreaks were reported in poultry in Italy. 258 of 287 poultry outbreaks detected in Europe were secondary outbreaks, suggesting that in the large majoryty of cases the spread of the virus was not due to wild birds.Allthe HPAI outbreaks were A(H5N8) apart from three,which were reported as A(H5N2) from Bulgaria. Genetic analysis of the HPAI A(H5N8) viruses isolated from the eastern and central European countries indicates that this is a reassortant between HPAI A(H5N8) viruses from Africa and LPAI viruses from Eurasia. Two distict subtypes were identified in Bulgaria, a novel reassortant A(H5N2) and A(H5N8) that is persisting in the country since 2016. There could be several reasons why only very few HPAI cases were detected in wild birds in this 2019‐2020 epidemic season and a better knowledge of wild bird movements and virus‐host interaction (e.g. susceptibility of the hosts to this virus) could help to understand the reasons for poor detection of HPAI infected wild birds. In comparison with the last reporting period, a decreasing number of HPAI A(H5)‐affected countries and outbreaks were reported from outside Europe. However, there is considerable uncertainty regarding the current epidemiological situation in many countries out of Europe. Four human cases due to A(H9N2) virus infection were reported during the reporting period from China.

Lifelong learning has deeply underpinned the resilience of biological organisms respect to a constantly changing environment. This flexibility has allowed the evolution of parallel-distributed systems able to merge past information with new stimulus for accurate and efficient brain-computation. Nowadays, there is a strong attempt to reproduce such intelligent systems in standard artificial neural networks (ANNs). However, despite some great results in specific tasks, ANNs still appear too rigid and static in real life respect to the biological systems. Thus, it is necessary to define a new neural paradigm capable of merging the lifelong resilience of biological organisms with the great accuracy of ANNs. Here, we present a digital implementation of a novel mixed supervised-unsupervised neural network capable of performing lifelong learning. The network uses a set of convolutional filters to extract features from the input images of the MNIST and the Fashion-MNIST training datasets. This information defines an original combination of responses of both trained classes and non-trained classes by transfer learning. The responses are then used in the subsequent unsupervised learning based on spike-timing dependent plasticity (STDP). This procedure allows the clustering of non-trained information thanks to bio-inspired algorithms such as neuronal redundancy and spike-frequency adaptation. We demonstrate the implementation of the neural network in a fully digital environment, such as the Xilinx Zynq-7000 System on Chip (SoC). We illustrate a user-friendly interface to test the network by choosing the number and the type of the non-trained classes, or drawing a custom pattern on a tablet. Finally, we propose a comparison of this work with networks based on memristive synaptic devices capable of continual learning, highlighting the main differences and capabilities respect to a fully digital approach.

Between 16 November 2019 and 15 February 2020, 36 highly pathogenic avian influenza (HPAI) A(H5N8) virus outbreakswere reported in Europe in poultry (n=34), captive birds (n=1) and wild birds (n=2), in Poland, Hungary, Slovakia, Romania, Germany, Czechiaand Ukraine,one HPAI outbreakcaused by a simultaneous infection with A(H5N2) and A(H5N8) was reported in poultry in Bulgaria, andtwo low pathogenic avian influenza (LPAI) A(H5) virus outbreaks were reported in poultryin the United Kingdom and in Denmark. Genomic characterisation of the HPAI A(H5N8) viruses suggests that they are reassortants of HPAI A(H5N8) viruses from Africa and LPAI viruses from Eurasia. It is likely that this reassortment occurred in wild migratory birds in Asia during the summer and then spread to eastern Europe with the autumnmigration. This is the first time that wild bird migration from Africa to Eurasia has been implicated in the long‐distance spread of HPAI viruses to the EU. Given the late incursion of HPAI A(H5N8) virus into the EU in this winter season (first outbreak reported on 30 December 2019), its overall restriction to eastern Europe, and the approaching spring migration, the risk of the virus spreadingfurther in the west via wild birds is decreasing for the coming months. Genetic analysis of the HPAI A(H5N2) and A(H5N8) viruses detected in the Bulgarian outbreak reveals that these virusesare both related to the 2018‐19 Bulgarian HPAI A(H5N8) viruses and not to the HPAI A(H5N8) viruses currently circulating in Europe.An increasing number of HPAI A(H5N1), A(H5N2), A(H5N5) and A(H5N6) virus outbreaks in poultry in Asia were reported during the time period for this report compared with the previous reporting period. Single outbreaks of HPAI A(H5N8) virus were notified by Saudi Arabia and South Africa. Furthermore, in contrast to the last report, HPAI virus‐positive wild birds were reported from Israel and one of the key migration areas in northern China.Two human cases due to A(H9N2) virus infection were reported during the reporting period.

Background: Smear microscopy is used to assess the patient's infectiousness at the time of initial diagnosis of pulmonary tuberculosis. However, its limited sensitivity and specificity highlights the need for new diagnostic strategies. The aim of our study was to assess the diagnostic accuracy of GX Ct value as a predictor of smear status and its usefulness to quantify mycobacterial load. Methods: All GX-positive sputum samples during a seven-year period were included in the study. Correlations among Ct values, smear status and TTD on liquid culture were calculated. An optimal Ct value for ruling in infectious patients was established. Clinical and radiological variables were also analyzed. Results: Sixty-eight samples from 65 patients were included. Ct value and TTD yielded a positive correlation (ρ = 0.714; p

Abstract Background The ability of Staphylococcus aureus to invade tissues and cause an infectious disease is the result of a multi-factorial process supported by the huge number of virulence factors inherent to this microorganism tightly regulated by the accessory gene regulator (agr). During antimicrobial therapy bacteria may be exposed to sub-inhibitory concentrations (subMICs) of antibiotics that may trigger transcriptional changes that may have an impact on the pathogenesis of infection. The objective of this study was to investigate the effect of oxacillin sub-MICs on agr system expression as the key component in the regulation of virulence in methicillin-susceptible (MSSA) and -resistant S. aureus (MRSA) strains. Furthermore, we studied the genetic basis of the agr locus and their potential association with the expression levels. Methods We have examined the expression of RNAIII and agrA mRNA as biomarkers for agr expression in the presence and absence of oxacillin subMICs in 10 MSSA and 4 MRSA clinical strains belonging to 5 clonal complexes (CC45-agrI, CC8-agrI, CC5-agrII, CC15-agrII and CC30-agrIII) causing endovascular complications. The DNA sequences of agr locus were obtained by whole genome sequencing. Results Our results revealed that exposure to subMICs of oxacillin had an impact on agr locus expression modifying the relative levels of expression with increases in 11 strains and with decreases in 3 strains. Thereby, the exposure to subMICs of oxacillin resulted in higher levels of expression of agr in CC15 and CC45 and lower levels in CC30. We also observed the presence of mutations in agrC and agrA in 13/14 strains with similar mutation profiles among strains within individual CCs except for strains of CC5. Although, agr expression levels differed among strains within CCs, the presence of these mutations was associated with differences in agr expression levels in most cases. Conclusions Changes in agr expression induced by exposure to oxacillin subMICs should be considered because they could lead to changes in the virulence modulation and have an adverse effect on the course of infection, especially in certain clonal complexes.

Between 16 February and 15 August 2019, five HPAI A(H5N8) outbreaks at poultry establishments in Bulgaria, two low pathogenic avian influenza (LPAI) A(H5N1) outbreaks in poultry in Denmark and one in captive birds in Germany, one LPAI A(H7N3) outbreak in poultry in Italy and one LPAI A(H7N7) outbreak in poultry in Denmark were reported in Europe. Genetic characterisation reveals that viruses from Denmark cluster with viruses previously identified in wild birds and poultry in Europe; while the Italian isolate clusters with LPAI viruses circulating in wild birds in Central Asia. No avian influenza outbreaks in wild birds were notified in Europe in the relevant period for this report. A decreased number of outbreaks in poultry and wild birds in Asia, Africa and the Middle East was reported during the time period for this report, particularly during the last three months. Furthermore, only six affected wild birds were reported in the relevant time period of this report. Currently there is no evidence of a new HPAI virus incursion from Asia into Europe. However, passive surveillance systems may not be sensitive for early detection if the prevalence or case fatality in wild birds is very low. Therefore, it is important to encourage and maintain passive surveillance in Europe encouraging a search for carcasses of wild bird species that are in the revised list of target species in order to detect any incursion of HPAI virus early and initiate warning. No human infections due to HPAI viruses ‐ detected in wild birds and poultry outbreaks in Europe ‐ have been reported during the last years and the risk of zoonotic transmission to the general public in Europe is considered very low.

Abstract This opinion updates the information included in the previous EFSA Scientific Opinion concerning the in planta control measures for Xylella fastidiosa, with a systematic review and critical analysis of the potential treatment solutions that have been published against this pest so far. The output of this opinion focuses on the application of chemical or biological treatments on living plants. In vitro studies, hot water treatments, use of resistant varieties and vector control are excluded from the review. The use of antibiotics is not considered due to the risk of antimicrobial resistance development. The use of weakly virulent or avirulent strains of X. fastidiosa is covered in this review, although this organism is an EU quarantine plant pest and its introduction in the EU territory is banned. Experiments were recently conducted to assess the effect of application of zinc, copper, and citric acid biocomplex, of N‐acetylcysteine, and of ‘diffusible signal factor’ (and of its homologs). Their results showed that these control measures were sometimes able to reduce symptoms caused by X. fastidiosa. Recent experiments also showed that several species of endophytic microorganisms, some bacteriophages and inoculation of weakly virulent/avirulent strains of X. fastidiosa could offer some protection against the Pierce's disease. However, based on the reviewed results, the Panel concludes that, although several published experiments show some effects in reducing symptoms development, the tested control measures are not able to completely eliminate X. fastidiosa from diseased plants. The Panel confirms as previously stated that there is currently no control measure available to eliminate the bacteria from a diseased plant in open field conditions.

No human infections due to highly pathogenic avian influenza (HPAI) A(H5N8) or A(H5N6) viruses ‐ detected in wild birds and poultry outbreaks in Europe ‐ have been reported so far and the risk of zoonotic transmission to the general public in Europe is considered very low. Between 16 November 2018 and 15 February 2019, two HPAI A(H5N8) outbreaks in poultry establishments in Bulgaria, two HPAI A(H5N6) outbreaks in wild birds in Denmark and one low pathogenic avian influenza (LPAI) A(H5N3) in captive birds in the Netherlands were reported in the European Union (EU). Genetic characterisation of the HPAI A(H5N6) viruses reveals that they cluster with the A(H5N6) viruses that have been circulating in Europe since December 2017. The wild bird species involved were birds of prey and were likely infected due to hunting or scavenging infected wild waterfowl. However, HPAI virus was not detected in other wild birds during this period. Outside the EU, two HPAI outbreaks were reported in poultry during the reporting period from western Russia. Sequence information on an HPAI A(H5N6) virus found in a common gull in western Russia in October 2018 suggests that the virus clusters within clade 2.3.4.4c and is closely related to viruses that transmitted zoonotically in China. An increasing number of outbreaks in poultry and wild birds in Asia, Africa and the Middle East was observed during the time period for this report. Currently there is no evidence of a new HPAI virus incursion from Asia into Europe. However, passive surveillance systems may not be sensitive enough if the prevalence or case fatality in wild birds is very low. Nevertheless, it is important to encourage and maintain a certain level of passive surveillance in Europe testing single sick or dead wild birds and birds of prey as they may be sensitive sentinel species for the presence of HPAI virus in the environment. A well‐targeted active surveillance might complement passive surveillance to collect information on HPAI infectious status of apparently healthy wild bird populations.

Migratory beekeeping is a widely extended practice aimed at increasing the yield of products and pollination services of honey bee colonies. However, it represents a stress factor, as it facilitates the dissemination of diseases and may compromise the genetic identity of the colonies involved. To analyze the extent of these effects, pathogens infestation rate and genetic composition were monitored in a field experiment comparing stationary and migratory colonies sharing the same environmental conditions but differing in management (stationary vs. migratory) and genetic background. We studied the pathogens infestation rate (Varroa destructor, Nosema spp., and Deformed Wing Virus (DWV)) at four different times: before migratory operation, two weeks later, at the end of the migratory period, and two weeks after the return of the migratory hives. An increased incidence of V. destructor and Nosema ceranae and a lower DWV viral load were found in migratory colonies. Temporary changes in genetic diversity were detected regardless of colony type, suggesting that stressors other than management affect the genetic diversity of the colonies. Our study demonstrates that migratory practices have variable effects on the health and genetic diversity of honey bee colonies, which should be taken into account for the development of sustainable beekeeping.

Wind power plants depend greatly on weather conditions, thus being considered intermittent, uncertain and non-dispatchable. Due to the massive integration of this energy resource in the recent decades, it is important that transmission and distribution system operators are able to model their electrical behaviour in terms of steady-state power flow, transient dynamic stability, and short-circuit currents. Consequently, in 2015, the International Electrotechnical Commission published Standard IEC 61400-27-1, which includes generic models for wind power generation in order to estimate the electrical characteristics of wind turbines at the connection point. This paper presents, describes and details the models for wind turbine topologies Types 1 and 2 following IEC 61400-27-1 for electrical simulation purposes, including the values for the parameters for the different subsystems. A hardware-in-the-loop combined with a real-time simulator is also used to analyse the response of such wind turbine topologies under voltage dips. The evolution of active and reactive powers is discussed, together with the wind turbine rotor and generator rotational speeds.

Between 16 August and 15 November 2018, 14 highly pathogenic avian influenza (HPAI) A(H5N8) outbreaks in poultry establishments in Bulgaria and seven HPAI A(H5N6) outbreaks, one in captive birds in Germany and six in wild birds in Denmark and the Netherlands were reported in the European Union (EU). No human infection due to HPAI A(H5N8) and A(H5N6) viruses have been reported in Europe so far. Seroconversion of people exposed during outbreaks in Russia has been reported in one study. Although the risk of zoonotic transmission to the general public in Europe is considered to be very low, appropriate personal protection measures of people exposed will reduce any potential risk. Genetic clustering of the viruses isolated from poultry in Bulgaria suggests three separate introductions in 2016 and a continuing circulation and transmission of these viruses within domestic ducks. Recent data from Bulgaria provides further indication that the sensitivity of passive surveillance of HPAI A(H5N8) in domestic ducks may be significantly compromised. Increased vigilance is needed especially during the periods of cold spells in winter when aggregations of wild birds and their movements towards areas with more favourable weather conditions may be encouraged. Two HPAI outbreaks in poultry were reported during this period from western Russia. Low numbers of HPAI outbreaks were observed in Africa and Asia, no HPAI cases were detected in wild birds in the time period relevant for this report. Although a few HPAI outbreaks were reported in Africa and Asia during the reporting period, the probability of HPAI virus introductions from non‐EU countries via wild birds particularly via the north‐eastern route from Russia is increasing, as the fall migration of wild birds from breeding and moulting sites to the wintering sites continues. Furthermore, the lower temperatures and ultraviolet radiation in winter can facilitate the environmental survival of any potential AI viruses introduced to Europe.

Bumblebees (genus Bombus Latreille) are pollinator insects of great ecological and economic importance, which commercial use for pollination has increased since the 80s. However, the introduction of foreign Bombus terrestris (Linnaeus) has resulted in a decline of native bumblebee populations in Japan, Chile or Argentina among others. To study the potential introgression of commercial B. terrestris into the Iberian endemic subspecies Bombus terrestris lusitanicus Krüger, it is necessary to find a precise molecular marker that differentiates both subspecies. For this purpose, comparative analyses were carried out between B. t. lusitanicus and B. t. terrestris (Linnaeus) from Spain and from Belgium by sequencing the nuclear genes elongation factor 1-α and arginine kinase and the mitochondrial gene 16S ribosomal RNA, and genotyping with eleven microsatellite loci. No differentiation was observed at the nuclear level, but haplotypes found within the 16S sequence correlated with the morphological characterization of the subspecies. In a case study including individuals sampled before the establishment of bumblebee rearing companies and others from recent samplings, we detected hybrid individuals (those with non-matching morphological subspecies and 16S haplotype) more frequently in the south supporting the naturalization of commercial B. t. terrestris and introgression events between both subspecies. This marker should be used in Iberian populations with the aim to support management and conservation actions in endemic populations of B. t. lusitanicus.

Staphylococcus aureus bacteremia (SAB) is associated with high morbidity and mortality, which varies depending on the source of infection. Nevertheless, the global molecular epidemiology of SAB and its possible association with specific virulence factors remains unclear. Using DNA microarrays, a total of 833 S. aureus strains (785 SAB and 48 colonizing strains) collected in Spain over a period of 15 years (2002–2017) were characterized to determine clonal complex (CC), agr type and repertoire of resistance and virulence genes in order to provide an epidemiological overview of CCs causing bloodstream infection, and to analyze possible associations between virulence genes and the most common sources of bacteremia. The results were also analyzed by acquisition (healthcare-associated [HA] and community-acquired [CA]), methicillin-resistant (MRSA) and methicillin-susceptible (MSSA) strains, and patient age (adults vs. children). Our results revealed high clonal diversity among SAB strains with up to 28 different CCs. The most prevalent CCs were CC5 (30.8%), CC30 (20.3%), CC45 (8.3%), CC8 (8.4%), CC15 (7.5%), and CC22 (5.9%), which together accounted for 80% of all cases. A higher proportion of CC5 was found among HA strains than CA strains (35.6 vs. 20.2%, p < 0.001). CC5 was associated with methicillin resistance (14.7 vs. 79.4%, p < 0.001), whereas CC30, CC45, and CC15 were correlated with MSSA strains (p < 0.001). Pathogen-related molecular markers significantly associated with a specific source of bacteremia included the presence of sea, undisrupted hlb and isaB genes with catheter-related bacteremia; sed, splE, and fib genes with endocarditis; undisrupted hlb with skin and soft tissue infections; and finally, CC5, msrA resistance gene and hla gene with osteoarticular source. Our study suggests an association between S. aureus genotype and place of acquisition, methicillin resistance and sources of bloodstream infection, and provides a valuable starting point for further research insights into intrinsic pathogenic mechanisms involved in the development of SAB.

Between 16 May and 15 August 2018, three highly pathogenic avian influenza (HPAI) A(H5N8) outbreaks in poultry establishments and three HPAI A(H5N6) outbreaks in wild birds were reported in Europe. Three low pathogenic avian influenza (LPAI) outbreaks were reported in three Member States. Few HPAI and LPAI bird cases have been detected in this period of the year, in accordance with the seasonal expected pattern of LPAI and HPAI. There is no evidence to date that HPAI A(H5N8) and A(H5N6) viruses circulating in Europe have caused any human infections. The risk of zoonotic transmission to the general public in Europe is considered to be very low. Several HPAI outbreaks in poultry were reported during this period from Russia. The presence of the A(H5N2) and A(H5N8) viruses in parts of Russia connected with fall migration routes of wild birds is of concern for possible introduction and spread with wild birds migrating to the EU. Although few AI outbreaks were observed in Africa, Asia and the Middle East during the reporting period, the probability of AI virus introductions from non‐EU countries via wild birds particularly via the north‐eastern route from Russia is increasing, as the fall migration of wild birds will start in the coming weeks. Further, the lower temperatures in autumn and winter may facilitate the environmental survival of avian influenza viruses potentially introduced to Europe.

Honey bee hives are moved yearly mainly for pollination, but also to take advantage of consecutive flowering events to get as many harvests of honey as possible and/or to find favorable sites for food sources and summer temperatures. Such movements may lead to pathogen spill-over with consequences on the honey bee health and finally on population decline. Ascosphaera apis is the causative agent of the chalkbrood disease, a pathology affecting honey bee larvae that significantly harms population growth and colony productivity. In this study, we detected the presence of A. apis in adult worker honey bees by PCR-amplification of the intergenic transcribed spacer (ITS1) of the ribosomal gene (rDNA). We first optimized the DNA extraction by testing different protocols in individual and pooled (colony level) adult honey bee samples. Subsequently, the presence of the fungus A. apis was assessed in both stationary and migratory colonies (subjected to small scale regional level movements) to determine the effect of migratory practices on the dispersal of this pathogen. Results confirmed a higher prevalence of A. apis in migratory apiaries when compared to stationary ones, indicating that migratory colonies are more likely to develop chalkbrood disease. Given these results, we suggest that beekeepers should be aware of the risks of pathogens spreading while moving beehives, even within a reduced geographic range.

Between 16 February and 15 May 2018, three highly pathogenic avian influenza (HPAI) A(H5N6) and 11 HPAI A(H5N8) outbreaks in poultry holdings, one HPAI A(H5N6) and one HPAI A(H5N8) outbreak in captive birds, and 55 HPAI A(H5N6) wild bird events were reported in Europe. There is no evidence to date that HPAI A(H5N6) viruses circulating in Europe are associated with clades infecting humans. Fewer HPAI wild bird cases have been detected than during the same period of previous year. Most of mortality events among wild birds involved single birds and species listed in the revised list of target species for passive surveillance. Raptor species constitute 74% of the HPAI‐infected wild birds found dead. Those raptor species probably became infected by hunting or scavenging HPAI virus‐positive birds, and so raptor cases may predominate later in the course of an HPAI epidemic. Despite the important HPAI virus incursion via wild birds there have been few associated HPAI A(H5N6) outbreaks in poultry. Fifteen low pathogenic avian influenza (LPAI) outbreaks were reported in three Member States. The risk of zoonotic transmission to the general public in Europe is considered to be very low. The situation in Africa and the Middle East should be closely monitored with regards to HPAI A(H5N1) and A(H5N8). Uncontrolled spread of the virus and subsequent further genetic evolution in regions geographically connected to Europe may increase uncertainty and the risk for further dissemination of virus. Long‐distance migrating wild birds from southern Africa, e.g. the common tern (Sterna hirundo), may be included in targeted active surveillance schemes at a few priority locations in Europe in order to detect HPAI A(H5)‐infected migrating birds early. However, the risk of HPAI introduction from non‐EU countries via migratory wild birds to Europe is still considered to be much lower for wild birds crossing the southern borders than for those crossing the north‐eastern borders.

In power systems with high wind energy penetration, the conjunction of wind power fluctuations and power system inertia reduction can lead to large frequency excursions, where the operating reserves of conventional power generation may be insufficient to restore the power balance. With the aim of evaluating the demand-side contribution to frequency control, a complete process to determine critical wind oscillations in power systems with high wind penetration is discussed and described in this paper. This process implies thousands of wind power series simulations, which have been carried out through a validated offshore wind farm model. A large number of different conditions have been taken into account, such as frequency dead bands, the percentages of controllable demand and seasonal factor influence on controllable loads. Relevant results and statistics are also included in the paper.

Moderate consumption of wine seems to produce positive health effects derived from the occurrence of bioactive polyphenols. The gut microbiota is involved in the metabolism of phenolic compounds, and these compounds and/or their metabolites may modulate gut microbiota through the stimulation of the growth of beneficial bacteria and the inhibition of pathogenic bacteria. The characterization of bacterial metabolites derived from polyphenols is essential in order to understand their effects, including microbial modulation, and therefore to associate dietary intake with particular health effects. This review aims to summarize the current information about the two-way “wine polyphenols–gut microbiota” interaction, from a perspective based on the experimental and analytical designs used. The availability of advanced methods for monitoring bacterial communities, along with the combination of in vitro and in vivo models, could help to assess the metabolism of polyphenols in the human body and to monitor total bacterial communities, and, therefore, to elucidate the implications of diet on the modulation of microbiota for delivering health benefits.

Abstract 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.

Staphylococcus aureus is a major cause of bacteremia and, even with appropriate clinical management, causes high morbidity, and mortality due to its involvement in endovascular complications and metastatic infections. Through different pathogenic in vivo and in vitro models we investigated the behavior of S. aureus most relevant clonal complexes (CCs) causing endovascular complications. We analyzed 14 S. aureus strains representing CC5, CC8, CC15, CC30, and CC45 that caused endovascular complications, including methicillin susceptible and resistant isolates and strains with different functionality of the agr global regulator. Their adherence to collagen, interaction with the endothelium, resistance to immune attack, capacity to form biofilm and virulence in the Galleria mellonella model were analyzed. CC30 and CC45 showed greater adhesion to collagen and CC8 showed a trend towards higher rate of intracellular persistence in endothelial cells. All CCs exhibited similar tolerance to neutrophil antimicrobial peptide hNP-1 and were capable of forming biofilms under static conditions. The virulence assay in the G. mellonella model demonstrated that CC15 and CC30 were the most and least virulent, respectively. The analysis of the genomic sequences of the most relevant virulence genes identified some CC15 specific gene patterns (absence of enterotoxins and sak gene) and variants (mainly in leucocidins and proteases), but did not reveal any gene or variant that could be responsible for the increased virulence detected for CC15 strains. Even though all the CCs were capable of causing endovascular complications, our results showed that different CCs are likely to produce these complications through different mechanisms which, if confirmed in more sophisticated models, would indicate the need to more specific management and therapeutic approaches.

Dietary polyphenols, including red wine phenolic compounds, are extensively metabolized during their passage through the gastrointestinal tract; and their biological effects at the gut level (i.e., anti-inflammatory activity, microbiota modulation, interaction with cells, among others) seem to be due more to their microbial-derived metabolites rather than to the original forms found in food. In an effort to improve our understanding of the biological effects that phenolic compounds exert at the gut level, this paper summarizes the changes observed in the human fecal metabolome after an intervention study consisting of a daily consumption of 250 mL of wine during four weeks by healthy volunteers (n = 33). It assembles data from two analytical approaches: (1) UPLC-ESI-MS/MS analysis of phenolic metabolites in fecal solutions (targeted analysis); and (2) UHPLC-TOF MS analysis of the fecal solutions (non-targeted analysis). Both approaches revealed statistically-significant changes in the concentration of several metabolites as a consequence of the wine intake. Similarity and complementarity between targeted and non-targeted approaches in the analysis of the fecal metabolome are discussed. Both strategies allowed the definition of a complex metabolic profile derived from wine intake. Likewise, the identification of endogenous markers could lead to new hypotheses to unravel the relationship between moderate wine consumption and the metabolic functionality of gut microbiota.

An alternative approach for combined frequency control in multi-area power systems with significant wind power plant integration is described and discussed in detail. Demand response is considered as a decentralized and distributed resource by incorporating innovative frequency-sensitive load controllers into certain thermostatically controlled loads. Wind power plants comprising variable speed wind turbines include an auxiliary frequency control loop contributing to increase total system inertia in a combined manner, which further improves the system frequency performance. Results for interconnected power systems show how the proposed control strategy substantially improves frequency stability and decreases peak frequency excursion (nadir) values. The total need for frequency regulation reserves is reduced as well. Moreover, the requirements to exchange power in multi-area scenarios are significantly decreased. Extensive simulations under power imbalance conditions for interconnected power systems are also presented in the paper.