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African swine fever (ASF) emerged in Germany in 2020. A few weeks after the initial occurrence, infected wild boar were detected in Saxony. In this study, data from wild boar surveillance in Saxony were analyzed. The analysis focused on the eastern districts of the state, where the disease likely spread through infected wild boar. Additionally, data from Meissen, located approximately 65 km further west, were examined. In Meissen, the disease emerged one year later, and a human-mediated introduction was suspected. To evaluate the progression of ASF in the two study areas over time, data from active and passive surveillance were analyzed both descriptively and using a Bayesian space–time model. Prevalence estimates were calculated for wild boar testing positive for the ASF virus and for ASF-specific antibodies. Higher prevalence estimates were observed in the eastern districts, indicating a stronger viral load, consistent with patterns seen in other European regions. Over time, seroprevalence increased, suggesting an accumulation of surviving wild boar. The findings provide important insights into the epidemiology of ASF in wild boar over time. They complement the existing knowledge and support targeted ASF control measures. This is particularly significant, as ASF continues to spread across Europe rather than being successfully eradicated.

Abstract Targeted search for wild boar carcasses is essential for successful control of African swine fever (ASF) in wild boar populations. To examine whether landscape conditions influence the probability of finding ASF-positive carcasses, this study analyzed Global Positioning System (GPS) coordinates of Latvian wild boar carcasses and hunted wild boar, extracted from the CSF/ASF wild boar surveillance database of the European Union, and random coordinates in Latvia. Geographic information system (GIS) software was used to determine the landscape type and landscape composition of carcass detection sites and to measure distances from the carcasses to nearest waterbodies, forest edges, roads and settlements. The results of the automated measurements were validated by manually analyzing a smaller sample. Wild boar carcasses were found predominantly in forested areas and closer to waterbodies and forest edges than random GPS coordinates in Latvia. Carcasses of ASF-infected wild boar were found more frequently in transitional zones between forest and woodland shrub, and at greater distances from roads and settlements compared to ASF-negative carcasses and random points. This leads to the hypothesis, that ASF-infected animals seek shelter in quiet areas further away from human disturbance. A detailed collection of information on the environment surrounding carcass detection sites is needed to characterize predilection sites more accurately.

The seasonality of African swine fever (ASF) outbreaks in domestic pigs differs between temperate and subtropical/tropical regions. We hypothesise that variations in the importance of wild boar-to-farm and farm-to-farm transmission routes shape these contrasting patterns, and we emphasise the implications for effective ASF control.

The implementation of management activities depends on both the attitude of the people performing the work and their understanding of why the work should be performed. In the context of animal husbandry, the implementation of such practices is crucial for the functionality of biosecurity. Therefore, it is important to know how farmers perceive biosecurity as a whole. An anonymous online survey was conducted among German farmers. In addition to general data about their farm, information about their existing concept of biosecurity, as well as about the assessment of possible introduction routes for animal diseases into the farm with regard to their likelihood, was gathered. Furthermore, information on measures to protect their farm against disease introduction were retrieved. Analysis showed that in general, farmers were aware of the importance of biosecurity and consequently had concepts of biosecurity on their farms. However, awareness about dangerous introduction routes for animal diseases into a farm was associated with a lack of knowledge of how to improve the measures in these areas. The role of the veterinarian in the context of biosecurity was highlighted and further problematic areas were indicated. Overall, the high level of commitment from farmers indicated a good implementation of daily practices.

Abstract Background African swine fever (ASF) has been present in Lithuania since 2014. The disease affects mainly the wild boar population. Thus, hunters play a key role in the performance of disease surveillance and control measures. We used participatory methods to gain insight into the knowledge of hunters and to include their perceptions in the design and the implementation of surveillance and control measures to increase their effectiveness. Results The willingness and the interest of hunters to participate was high, but only eight focus group meetings with 33 hunters could be held due to the COVID-19 pandemic. The overall knowledge of Lithuanian hunters regarding ASF, investigated by semi-structured interviews, was sufficient to understand their part in ASF control and surveillance. However, their knowledge did not necessarily lead to an increased acceptance of some ASF control measures, like the targeted hunting of female wild boar. Participating hunters showed a good understanding of the processes of the surveillance system. Their trust in the performance within this system was highest towards the hunters themselves, thus emphasizing the importance of acknowledging their role in the system. Hunters refused measures including the reduction of hunting activities. They feared a complete elimination of the wild boar population, which in turn demonstrates the necessity to increase professional information exchange. Conclusions The perceptions of Lithuanian hunters regarding ASF surveillance and control in wild boar resembled those obtained in neighboring countries. It is imperative to communicate the results with decision-makers, to consider the views of hunters, when designing or adapting measures to control ASF in wild boar and to communicate with hunters on these measures and their justification.

African swine fever (ASF) has posed a significant threat to Ukrainian pig farming since its identification in 2012. In this study, recognising the pivotal role of pigkeepers in disease control, we conducted ten focus groups involving 52 smallholders across eight regions in Ukraine. Using participatory methods, we revealed their awareness of ASF signs, transmission routes, preventive measures, and the perceptions of stakeholders involved in ASF control. Furthermore, we identified the smallholders’ acceptance of eradication and restriction measures, the perceived impact of zoning consequences, and their main sources of ASF information. Smallholders identified fever and skin haemorrhage as the most indicative signs of ASF and highlighted rodents as a primary transmission concern. Disinfection was seen as the most effective measure for preventing the introduction of ASF. Pigkeepers who perceived their stakeholder role in ASF control showed more trust in themselves and veterinarians than in central veterinary authorities. Farm-level ASF eradication measures were generally accepted; however, culling within the protection zone was least accepted, with economic losses listed as the most impactful consequence for pigkeepers. For ASF information, pigkeepers favour web searches and veterinarians, as well as traditional media and word-of-mouth news. This study provides valuable insights into refining the ASF communication strategies in Ukraine.

Since the first occurrence of African swine fever (ASF) in wild boar in Germany in 2020, the disease has primarily affected the wild boar population in the eastern part of the country close to the border with Poland. Local hunters play a crucial role in implementing surveillance and control. To evaluate their perceptions of existing control measures and analyze regional differences between hunters from ASF-affected and non-affected regions, a questionnaire study was conducted among the German hunting community. Hunters from non-affected areas held a more optimistic view regarding the effectiveness of control measures compared to hunters from affected areas. However, control measures that hinder hunting were generally perceived as ineffective. Measures that collided with hunters’ understanding of fair hunting practices were regarded as controversial. Financial incentives and reducing bureaucracy were the most favored approaches to increase hunters’ participation. Moreover, the possibility of eating or selling the meat of hunted wild boar and the provision of infrastructure for implementing ASF control were considered motivating. Thus, this study highlights the importance of compensating hunters and addressing their concerns to maintain their engagement in ASF control. To enhance compliance with controversial measures, thoughtful communication and raising awareness are essential.

Since 2007, African swine fever (ASF) has spread widely within Europe and beyond. Most affected countries recorded outbreaks in domestic pigs and cases in wild boar. Outbreak data from 2014 to 2021 were used to investigate the seasonal pattern of ASF in domestic pigs and wild boar across affected member states of the European Union, since knowledge of seasonal patterns may provide the potential to adapt prevention, surveillance and control during times of increased risk. In domestic pigs, a yearly peak was observed in many European countries in summer (predominantly in July and August). In wild boar, the patterns showed more variability. In many countries, there was a seasonal peak of ASF occurrence in winter (predominantly in January and December), with an additional summer peak in the Baltic States (predominantly in July) and a further spring peak in Poland (predominantly in March). The observed seasonal effects may be related to the abundance and population dynamics of wild boar and to seasonality in pig farming. Moreover, ASF occurrence may also be influenced by human activities in both domestic pigs and wild boar.

Abstract The first cases of African swine fever (ASF) were detected in the Lithuanian wild boar population in 2014. Since then, the disease spread slowly through the whole country, affecting both, wild boar and domestic pigs. In the other Baltic states, which both are also affected by ASF since 2014, the recent course of ASF prevalence suggests that the countries might be well under way of disease elimination. In contrast, in Lithuania the epidemic seems to be still in full progress. In the present study, we aimed to extend a previous prevalence study in Lithuania. Looking at ASF virus (ASFV) and seroprevalence estimates of wild boar in all months of 2018 and in all affected municipalities in Lithuania, the course of ASF was evaluated on a temporal and spatial scale. A non-spatial beta-binomial model was used to correct for under- or overestimation of the average prevalence estimates. Within 2018 no big differences between the prevalence estimates were seen over time. Despite of the lower sample size, highest ASFV prevalence estimates were found in dead wild boar, suggesting higher detection rates through passive surveillance than through active surveillance. Accordingly, with the maximum prevalence of 87.5% in May 2018, the ASFV prevalence estimates were very high in wild boar found dead. The number of samples originating from hunted animals (active surveillance) predominated clearly. However, the ASFV prevalence in those animals was lower with a maximum value of 2.1%, emphasizing the high value of passive surveillance. A slight increase of the seroprevalence in hunted wild boar could be seen over time. In the center of Lithuania, a cluster of municipalities with high ASFV and seroprevalence estimates was found. The results of the study indicate that ASFV is still circulating within the Lithuanian wild boar population, constituting a permanent risk of disease transmission into domestic pig holdings. However, additional, more recent data analyses are necessary to re-evaluate the course of ASF in Lithuania and thus, to be able to make a statement about the stage of the ASF epidemic in the country. This is of huge importance for Lithuania for evaluating control measures and their efficacy, but also for neighbouring countries to assess the risk of disease spread from Lithuania.

African swine fever (ASF) emerged in Latvia in 2014. In 2020, the virus has been detected in the German federal state, Saxony. In both regions, the virus was probably introduced by infected wild boar coming from affected neighboring countries. As the current ASF control strategy at EU level had not yet been developed at the time of ASF introduction into Latvia, disease control measures in both study areas differed over time. Assessing the course of ASF in Saxony and the implemented control strategies, the first 18 months of the ASF epidemic were epidemiologically compared between Saxony and Latvia. ASF wild boar surveillance data were analyzed and the prevalence of ASF virus-positive wild boar was estimated. For estimating the wild boar density, the numbers of wild boar per km² were calculated for the respective geographical areas. The number of samples collected from hunted wild boar and wild boar found dead was higher in Saxony. The ASF virus prevalence in Latvia was significantly higher than in Saxony, indicating that Saxony has had more time for getting prepared for dealing with an ASF incursion. Experience from other countries and the rapid implementation of new control strategies may have helped Saxony deal with ASF.

African swine fever (ASF) is an internationally-spreading viral pig disease that severely damages agricultural pork production and trade economy as well as social welfare in disease-affected regions. A comprehensive understanding of ASF risk factors is imperative for efficient disease control. As the absence of effective ASF vaccines limits disease management options, the identification and minimisation of ASF-associated risk factors is critical to preventing ASF outbreaks. Here, we compile currently known potential ASF risk factors identified through a systematic literature review. We found 154 observation-based and 1239 potential ASF risk factors, which we were able to group into the following defined risk categories: ‘ASF-virus’, ‘Biosecurity’, ‘Disease control’, ‘Environment’, ‘Husbandry’, ‘Movement’, ‘Network’, ‘Pig’, ‘Society’ and ‘Surveillance’. Throughout the epidemiological history of ASF there have been similar risk categories, such as ‘Environment’-related risk factors, predominantly reported in the literature irrespective of the ASF situation at the time. While ASF risk factor reporting has markedly increased since 2010, the majority of identified risk factors overall have referred to domestic pigs. The reporting of risk factors for ASF in wild boar mostly commenced from 2016 onwards. The compendium of ASF risk factors presented herein defines our current knowledge of ASF risk factors, and critically informs ASF-related problem solving.

Since the first detected African swine fever (ASF) cases in Lithuanian wild boar in 2014, the virus has occurred in many other member states of the European Union (EU), most recently in Belgium in 2018 and in Germany in 2020. Passive surveillance and various control measures are implemented as part of the strategy to stop disease spread in the wild boar population. Within this framework, hunters perform important activities, such as the removal of carcasses, fencing or hunting. Therefore, the successful implementation of these measures largely depends on their acceptability by hunters. Methods of participatory epidemiology can be used to determine the acceptance of control measures. The use of participatory methods allows the involvement of key stakeholders in the design, the implementation and the analysis of control and surveillance activities. In the present study, two studies that had been conducted using participatory epidemiology with hunters in Estonia and Latvia were compared on the topics recruitment, participants, facilitators, focus group discussion (FGDs) and their contents. The aim was to evaluate similarities and differences in the two studies and to identify a broader spectrum of possibilities to increase the willingness of hunters supporting the fight against ASF. Evaluating all conducted FGDs in both countries showed primarily similarities in the perceptions and opinions of the hunters in Estonia and Latvia. One notable difference was that passive surveillance in Latvia was perceived mostly as topic of duty and ethics rather than an issue driven by incentives. Participatory methods have proven to be an effective tool in the evaluation of the acceptance of established ASF control systems. The results of this study point out further chances for improving the cooperation with hunters in the future. Nevertheless, the importance of gathering and analyzing the opinions of hunters in all ASF affected countries individually is highlighted.

African swine fever (ASF) was first detected in Lithuania, Latvia, and Estonia in 2014 and has since been circulating in the Baltic States with a similar epidemiological course characterized by persistence of the disease in the wild boar population and occasional spill-over infections in domestic pigs. The aim of the present study was to evaluate surveillance data on ASF in wild boar from the three countries to improve our understanding of the course of the disease. ASF surveillance and wild boar population data of the countries were analyzed. In all three countries, a decrease in the prevalence of ASF virus-positive wild boar was observed over time. Although somewhat delayed, an increase in the seroprevalence was seen. At the same time, the wild boar population density decreased significantly. Towards the end of the study period, the wild boar population recovered, and the prevalence of ASF virus-positive wild boar increased again, whereas the seroprevalence decreased. The decreasing virus prevalence has obviously led to virus circulation at a very low level. Together with the decreasing wild boar population density, the detection of ASF-infected wild boar and thus ASF control has become increasingly difficult. The course of ASF and its continuous spread clearly demonstrate the necessity to scrutinize current ASF surveillance and control strategies fundamentally and to consider new transdisciplinary approaches.

While numerous risk factors of African swine fever (ASF) transmission to domestic pigs have been described, ASF virus introduction has often not been traced back to one single defined cause. The large number of ASF outbreaks that occurred in domestic pigs in Lithuania from 2014 through to 2018 raised the question regarding whether outbreak-specific risk factors and transmission routes could be identified. Therefore, a prospective matched case-control study was designed. Data from 18 outbreaks that occurred in Lithuanian in 2019 and 36 control farms were analyzed. Conditional multivariable logistic regression showed that two or more visits by veterinary inspection of a farm had a significant preventive effect on the occurrence of ASF on a farm (Odds ratio (OR) 14.21, confidence interval (CI) 1.09–185.60 for farms not inspected vs. farms inspected twice or more a year), while certain practices (e.g., mushroom picking, sharing equipment, etc.), which might facilitate the indirect introduction of ASF from fields and forests into piggeries, significantly increased the odds of an outbreak (OR 5.18, CI 1.10–24.44). The results of the study highlight the importance of veterinary inspections for increasing the biosecurity level on pig farms and the awareness of ASF. The knowledge on potential protective and risk factors may help to improve the prevention and control of ASF outbreaks in domestic pig farms in Lithuania and other affected countries.

African swine fever (ASF) emerged in Estonia in 2014. From February 2019 to August 2020, no pigs or wild boar tested positive for ASF virus (ASFV), only ASFV-specific antibodies could be detected in shot wild boar. However, ASF recently re-emerged in wild boar. We tested three hypotheses that might explain the current situation: (i) ASFV may have been present throughout, but at a prevalence below the detection limit; (ii) seropositive wild boar may have remained infectious (i.e., virus-carriers) and kept the epidemic going; or (iii) ASF was gone for 1.5 years, but was recently re-introduced. Using Estonian surveillance data, the sensitivity of the surveillance system and the confidence in freedom from ASF were estimated. Furthermore, the detection probability was determined and cluster analyses were performed to investigate the role of serological positive wild boar. The results suggest that the surveillance system was not able to detect virus circulation at a design prevalence below 1%. With respect to the confidence in freedom from ASF, the results indicate that circulating virus should have been detected over time, if the prevalence was ≥2%. However, the decreasing wild boar population density and ongoing surveillance activities made ASFV circulation at a low prevalence unlikely. Cluster analyses provided no evidence for a significant accumulation of serologically positive wild boar in temporal connection to the re-emergence of ASFV. Further targeted research, such as long-term experimental studies and molecular epidemiology, is necessary to improve our knowledge on the epidemiology of ASF and to control the disease more effectively.

A detailed understanding of environmental risk factors for African swine fever (ASF) in wild boar will be not only essential for risk assessments but also for timely and spatially informed allocation of resources in order to manage wild boar-targeted ASF control measures efficiently. Here, we review currently known environmental risk factors that can influence the occurrence of ASF virus infection in wild boar when compared to disease occurrence in wild boar of a non-exposed reference scenario. Accordingly, the exposure of wild boar to environmental risk factors related to (1) climate, (2) land cover, (3) human activity, (4) wild boar and (5) ASF were evaluated. As key environmental risk factors in this review, increased ASF occurrence in wild boar was associated with seasonal patterns, forest coverage, presence of water, human presence, farming activities, wild boar density and ASF nearness. The review highlights inconsistencies in some of these risk factor associations with disease detection in space and time and may provide valuable insights for the investigation of ASF transmission dynamics. The examined risk information was applied to consider potential improvements of the ASF control strategy in wild boar regarding disease surveillance, hunting, wild boar carcass searches and ASF barrier implementation.

Abstract For the global pig industry, classical (CSF) and African swine fever (ASF) outbreaks are a constantly feared threat. Except for Sardinia, ASF was eradicated in Europe in the late 1990s, which led to a research focus on CSF because this disease continued to be present. However, ASF remerged in eastern Europe in 2007 and the interest in the disease, its control and epidemiology increased tremendously. The similar names and the same susceptible species suggest a similarity of the two viral diseases, a related biological behaviour and, correspondingly, similar epidemiological features. However, there are several essential differences between both diseases, which need to be considered for the design of control or preventive measures. In the present review, we aimed to collate differences and similarities of the two diseases that impact epidemiology and thus the necessary control actions. Our objective was to discuss critically, if and to which extent the current knowledge can be transferred from one disease to the other and where new findings should lead to a critical review of measures relating to the prevention, control and surveillance of ASF and CSF. Another intention was to identify research gaps, which need to be closed to increase the chances of a successful eradication of ASF and therefore for a decrease of the economic threat for pig holdings and the international trade.

The introduction of genotype II African swine fever (ASF) virus, presumably from Africa into Georgia in 2007, and its continuous spread through Europe and Asia as a panzootic disease of suids, continues to have a huge socio-economic impact. ASF is characterized by hemorrhagic fever leading to a high case/fatality ratio in pigs. In Europe, wild boar are especially affected. This review summarizes the currently available knowledge on ASF in wild boar in Europe. The current ASF panzootic is characterized by self-sustaining cycles of infection in the wild boar population. Spill-over and spill-back events occur from wild boar to domestic pigs and vice versa. The social structure of wild boar populations and the spatial behavior of the animals, a variety of ASF virus (ASFV) transmission mechanisms and persistence in the environment complicate the modeling of the disease. Control measures focus on the detection and removal of wild boar carcasses, in which ASFV can remain infectious for months. Further measures include the reduction in wild boar density and the limitation of wild boar movements through fences. Using these measures, the Czech Republic and Belgium succeeded in eliminating ASF in their territories, while the disease spread in others. So far, no vaccine is available to protect wild boar or domestic pigs reliably against ASF.

African swine fever (ASF) has been present in Lithuania since 2014. It is mainly the wild boar population that is affected. Currently, little is known about the epidemiological course of ASF in Lithuania. In the present study, ASF surveillance data from 2016–2021 were analyzed. The numbers of samples taken from hunted wild boar and wild boar found dead per year and month were recorded and the prevalence was estimated for each study month and administrative unit. A Bayesian space–time model was used to calculate the temporal trend of the prevalence estimates. In addition, population data were analyzed on a yearly basis. Most samples were investigated in 2016 and 2017 and originated from hunted animals. Prevalence estimates of ASF virus-positive wild boar decreased from May 2019 onwards. Seroprevalence estimates showed a slight decrease at the same time, but they increased again at the end of the study period. A significant decrease in the population density was observed over time. The results of the study show that ASF is still present in the Lithuanian wild boar population. A joint interdisciplinary effort is needed to identify weaknesses in the control of ASF in Lithuania and to combat the disease more successfully.

After the introduction of African swine fever (ASF) into Lithuania in 2014, continuous spread of the disease resulted in infection of the wild boar populations in most parts of Lithuania. The virus has been moving closer to other Western European countries where pig density is high. An efficient surveillance system detecting ASF cases early in domestic and wild animals is necessary to manage this disease. To make surveillance appropriate and effective, it is critical to understand how key players perceive the implemented control measures. This study investigated the attitudes and beliefs of hunters in Lithuania regarding currently implemented or proposed measures for the control of ASF in the wild boar population. Study data were collected through questionnaires distributed via the internet and by hunting associations in Lithuania. In total, 621 fully completed questionnaires were received and analyzed. All measures interfering with extensive hunting, like ban of driven or individual hunting or ban of supplementary feeding were considered as unacceptable and as ineffective measures to control ASF in wild boar. However, selective hunting of female wild boar was generally considered as an unethical act and therefore rejected. Some measures that seem to have been successful in other countries, like involvement of additional forces, were rejected by Lithuanian hunters, thus implementation of these measures could be difficult. The study highlighted that there is a need for improving important relationships with other stakeholders, since many hunters expressed a lack of trust in governmental institutions and regarded cooperation with them as insufficient. Hunters emphasized that their motivation to support passive surveillance measures could be improved with financial compensation and reduction of workload. The present study provides insights into hunters’ perceptions, which may be used as a foundation for additional discussions with these important stakeholders and for adapting measures to improve their acceptance if appropriate.

Searching for the term “African swine fever” (ASF) in the title of publications in the Web of Science, PubMed and Scopus during the period of 1955–2020 resulted in an average of 2009 detected articles [...]

Estonia has been combatting African swine fever (ASF) for six years now. Since October 2017, the disease has only been detected in the wild boar population, but trade restrictions had to remain in place due to international regulations. Yet, the epidemiological course of the disease has changed within the last few years. The prevalence of ASF virus (ASFV)-positive wild boar decreased steadily towards 0%. In February 2019, the last ASFV-positive wild boar was detected. Since then, positive wild boar samples have exclusively been positive for ASFV-specific antibodies, suggesting the possible absence of circulating ASFV in the Estonian wild boar population. However, as the role of seropositive animals is controversially discussed and the presence of antibody-carriers is regarded as an indication of virus circulation at EU and OIE level, Estonia remains under trade restrictions. To make the disease status of a country reliable for trading partners and to facilitate the process of declaration of disease freedom, we suggest to monitor the prevalence of seropositive wild boar in absence of ASFV-positive animals. The possibility to include ASF in the list of diseases, for which an official pathway for recognition of disease status is defined by the OIE should be evaluated.

The Encyclopedia of Life (EOL, http://eol.org) aims to provide unprecedented global access to a broad range of information about life on Earth. It currently contains 3.5 million distinct pages for taxa and provides content for 1.3 million of those pages. The content is primarily contributed by EOL content partners (providers) that have a more limited geographic, taxonomic or topical scope. EOL aggregates these data and automatically integrates them based on associated scientific names and other classification information. EOL also provides interfaces for curation and direct content addition. All materials in EOL are either in the public domain or licensed under a Creative Commons license. In addition to the web interface, EOL is also accessible through an Application Programming Interface. In this paper, we review recent developments added for Version 2 of the web site and subsequent releases through Version 2.2, which have made EOL more engaging, personal, accessible and internationalizable. We outline the core features and technical architecture of the system. We summarize milestones achieved so far by EOL to present results of the current system implementation and establish benchmarks upon which to judge future improvements. We have shown that it is possible to successfully integrate large amounts of descriptive biodiversity data from diverse sources into a robust, standards-based, dynamic, and scalable infrastructure. Increasing global participation and the emergence of EOL-powered applications demonstrate that EOL is becoming a significant resource for anyone interested in biological diversity.

Since the introduction of African swine fever (ASF) into Georgia in 2007, the disease has been spreading in an unprecedented way. Many countries that are still free from the disease fear the emergence of ASF in their territory either in domestic pigs or in wild boar. In the past, ASF was often described as being a highly contagious disease with mortality often up to 100%. However, the belief that the disease might enter a naïve population and rapidly affect the entire susceptible population needs to be critically reviewed. The current ASF epidemic in wild boar, but also the course of ASF within outbreaks in domestic pig holdings, suggest a constant, but relatively slow spread. Moreover, the results of several experimental and field studies support the impression that the spread of ASF is not always fast. ASF spread and its speed depend on various factors concerning the host, the virus, and also the environment. Many of these factors and their effects are not fully understood. For this review, we collated published information regarding the spreading speed of ASF and the factors that are deemed to influence the speed of ASF spread and tried to clarify some issues and open questions in this respect.

Introduction Neoadjuvant chemoimmunotherapy has achieved overall survival (OS) benefit for patients with resectable non-small cell lung cancer (NSCLC). Here, we present outcomes after 3 years of follow-up from the first reported study of neoadjuvant atezolizumab+chemotherapy.Methods This open-label, multicenter single-arm investigator-initiated phase II study conducted at three US hospitals tested up to four cycles of atezolizumab, carboplatin, and nab-paclitaxel prior to surgery. Major pathological response (MPR, primary endpoint) was previously reported; here, we report 3-year disease-free survival (DFS), OS, and clinical characteristics of patients developing brain metastases (BM) with integrated data from tumor genomics, gene expression, and quantitative immunofluorescent measurement of immune markers.Results Of 30 enrolled patients, 29 were taken to the operating room. 26 underwent R0 resection, with 17 experiencing MPR (10 pCR). With a median follow-up of 39.5 months, the median OS was 55.8 months, and the median DFS was 34.5 months. Landmark OS at 36 months was 77%. Among 14 patients with recurrent disease, 6 patients had BM. Patients whose tumors had mutations in STK11 and KEAP1 did not have a significantly higher incidence of BM. Reduced copy number of STK11 and KEAP1, both residing on chromosome 19p, was observed in ~1/3 of tumors. Reduced CN of STK11 was significantly associated with worse pathological response and incidence of BM.Conclusions Consistent with recent phase III studies, 3-year OS data with neoadjuvant atezolizumab+chemotherapy was associated with prolonged PFS and OS. Establishing associations between STK11 and KEAP1 genomic alterations and key clinical outcomes in early-stage NSCLC requires further study.

The recent proliferation of Extended Reality (XR) applications in geoscience education and research has opened new avenues for the enhanced visualization and analysis of the Earth’s geodata. This study specifically explores the benefits for teaching when supplementing industry standard software packages, such as Paleoscan, Petrel, and JewelSuite, with 3D visualization in XR. The teaching focuses on but is not limited to an understanding of subsurface seismic and well data. During this study, the undergraduate Petroleum Geoscience students transitioned from 2D computer screen visualizations to immersive XR tools. The dataset selected for teaching focuses on the subsurface carbonate EX field in the South China Sea. The EX-field in Central Luconia is located 100–300 km from Sarawak’s coastline in water depths of 60–140 m. It includes a post-stacked time- 3D seismic cube linked to wells, allowing students to work with seismic data, adjust scales, and conduct preliminary seismic analysis. The findings revealed a significant improvement in respondents’ skills in comprehending and analysing seismic and core data, enhancing the overall learning experience in Petroleum Geoscience. This paper also examines the students’ feedback on their learning experiences during virtual subsurface visualization throughout their university degree in geoscience. For evaluating learning success, we used an approach that merges quantitative and qualitative data, The students’ perceptions were assessed through anonymous quantitative surveys and questions. The analysis of student responses emphasizes the valuable learning experience offered by 3D virtual environments designed for realistic first-person navigation and freedom of movement, like a real field experience. The results highlight the potential of virtual subsurface visualization for imparting essential skills to geosciences.

Summary: Stem cells (SCs) are critical to maintain tissue homeostasis. However, it is currently not known whether signaling through cell junctions protects quiescent epithelial SC reservoirs from depletion during disease-inflicted damage. Using the autoimmune model disease pemphigus vulgaris (PV), this study reveals an unprecedented role for a desmosomal cadherin in governing SC quiescence and regeneration through adhesion signaling in the multipotent mouse hair follicle compartment known as the bulge. Autoantibody-mediated, mechanical uncoupling of desmoglein (Dsg) 3 transadhesion activates quiescent bulge SC which lose their multipotency and stemness, become actively cycling, and finally delaminate from their epithelial niche. This then initiates a self-organized regenerative program which restores Dsg3 function and bulge morphology including SC quiescence and multipotency. These profound changes are triggered by the sole loss of functional Dsg3, resemble major signaling events in Dsg3−/− mice, and are driven by SC-relevant EGFR activation and Wnt modulation requiring longitudinal repression of Hedgehog signaling.

Abstract Background Microscopic examination of human blood samples is an excellent opportunity to assess general health status and diagnose diseases. Conventional blood tests are performed in medical laboratories by specialized professionals and are time and labor intensive. The development of a point-of-care system based on a mobile microscope and powerful algorithms would be beneficial for providing care directly at the patient's bedside. For this purpose human blood samples were visualized using a low-cost mobile microscope, an ocular camera and a smartphone. Training and optimisation of different deep learning methods for instance segmentation are used to detect and count the different blood cells. The accuracy of the results is assessed using quantitative and qualitative evaluation standards. Results Instance segmentation models such as Mask R-CNN, Mask Scoring R-CNN, D2Det and YOLACT were trained and optimised for the detection and classification of all blood cell types. These networks were not designed to detect very small objects in large numbers, so extensive modifications were necessary. Thus, segmentation of all blood cell types and their classification was feasible with great accuracy: qualitatively evaluated, mean average precision of 0.57 and mean average recall of 0.61 are achieved for all blood cell types. Quantitatively, 93% of ground truth blood cells can be detected. Conclusions Mobile blood testing as a point-of-care system can be performed with diagnostic accuracy using deep learning methods. In the future, this application could enable very fast, cheap, location- and knowledge-independent patient care.

Introduction: Whereas tumor biopsy is the reference standard for genomic profiling of advanced NSCLC, there are now multiple assays approved by the Food and Drug Administration for liquid biopsy testing of circulating tumor DNA. Here, we study the incremental value that liquid biopsy comprehensive genomic profiling (CGP) adds to tissue molecular testing. Methods: Patients with metastatic NSCLC were enrolled in a prospective diagnostic study to receive circulating tumor DNA CGP; tissue CGP was optional in addition to their standard tissue testing. Focusing on nine genes listed per the National Comprehensive Cancer Network (NCCN) guidelines, liquid CGP was compared with available tissue testing results across three subcohorts: tissue CGP, standard-of-care testing of up to five biomarkers, or no tissue testing. Results: A total of 515 patients with advanced nonsquamous NSCLC received liquid CGP. Among 131 with tissue CGP results, NCCN biomarkers were detected in 86 (66%) with tissue CGP and 56 (43%) with liquid CGP (p < 0.001). Adding liquid CGP to tissue CGP detected no additional patients with NCCN biomarkers, whereas tissue CGP detected NCCN biomarkers in 30 patients (23%) missed by liquid CGP. Studying 264 patients receiving tissue testing of up to five genes, 102 (39%) had NCCN biomarkers detected in tissue, with an additional 48 (18%) detected using liquid CGP, including 18 with RET, MET, or ERBB2 drivers not studied in tissue. Conclusions: For the detection of patients with advanced nonsquamous NSCLC harboring 9 NCCN biomarkers, liquid CGP increases detection in patients with limited tissue results, but does not increase detection in patients with tissue CGP results available. In contrast, tissue CGP can add meaningfully to liquid CGP for detection of NCCN biomarkers and should be considered as a follow-up when an oncogenic driver is not identified by liquid biopsy.

Background Non-small cell lung cancer (NSCLC) patients bearing targetable oncogene alterations typically derive limited benefit from immune checkpoint blockade (ICB), which has been attributed to low tumor mutation burden (TMB) and/or PD-L1 levels. We investigated oncogene-specific differences in these markers and clinical outcome.Methods Three cohorts of NSCLC patients with oncogene alterations (n=4189 total) were analyzed. Two clinical cohorts of advanced NSCLC patients treated with ICB monotherapy [MD Anderson (MDACC; n=172) and Flatiron Health-Foundation Medicine Clinico-Genomic Database (CGDB; n=894 patients)] were analyzed for clinical outcome. The FMI biomarker cohort (n=4017) was used to assess the association of oncogene alterations with TMB and PD-L1 expression.Results High PD-L1 expression (PD-L1 ≥50%) rate was 19%–20% in classic EGFR, EGFR exon 20 and HER2-mutant tumors, and 34%–55% in tumors with ALK, BRAF V600E, ROS1, RET, or MET alterations. Compared with KRAS-mutant tumors, BRAF non-V600E group had higher TMB (9.6 vs KRAS 7.8 mutations/Mb, p=0.003), while all other oncogene groups had lower TMB (p

BackgroundIn non-small cell lung cancer (NSCLC), PD-L1 expression on either tumor cells (TC) or both TC and tumor-infiltrating immune cells (IC) is currently the most used biomarker in cancer immunotherapy. However, the mechanisms involved in PD-L1 regulation are not fully understood. To provide better insight in these mechanisms, a multiangular analysis approach was used to combine protein and mRNA expression with several clinicopathological characteristics.Patients and methodsArchival tissues from 640 early stage, resected NSCLC patients were analyzed with immunohistochemistry for expression of PD-L1 and CD8 infiltration. In addition, mutational status and expression of a selection of immune genes involved in the PD-L1/PD-1 axis and T-cell response was determined.ResultsTumors with high PD-L1 expression on TC or on IC represent two subsets of NSCLC with minimal overlap. We observed that PD-L1 expression on IC irrespective of expression on TC is a good marker for inflammation within tumors. In the tumors with the highest IC expression and absent TC expression an association with reduced IFNγ downstream signaling in tumor cells was observed.ConclusionsThese results show that PD-L1 expression on TC and IC are both independent hallmarks of the inflamed phenotype in NSCLC, and TC-negative/IC-high tumors can also be categorized as inflamed. The lack of correlation between PD-L1 TC and IC expression in this subgroup may be caused by impaired IFNγ signaling in tumor cells. These findings may bring a better understanding of the tumor-immune system interaction and the clinical relevance of PD-L1 expression on IC irrespective of PD-L1 expression on TC.

Automatic image reconstruction is critical to cope with steadily increasing data from advanced microscopy. We describe here the Fiji macro 3D ART VeSElecT which we developed to study synaptic vesicles in electron tomograms. We apply this tool to quantify vesicle properties (i) in embryonic Danio rerio 4 and 8 days past fertilization (dpf) and (ii) to compare Caenorhabditis elegans N2 neuromuscular junctions (NMJ) wild-type and its septin mutant (unc-59(e261)). We demonstrate development-specific and mutant-specific changes in synaptic vesicle pools in both models. We confirm the functionality of our macro by applying our 3D ART VeSElecT on zebrafish NMJ showing smaller vesicles in 8 dpf embryos then 4 dpf, which was validated by manual reconstruction of the vesicle pool. Furthermore, we analyze the impact of C. elegans septin mutant unc-59(e261) on vesicle pool formation and vesicle size. Automated vesicle registration and characterization was implemented in Fiji as two macros (registration and measurement). This flexible arrangement allows in particular reducing false positives by an optional manual revision step. Preprocessing and contrast enhancement work on image-stacks of 1nm/pixel in x and y direction. Semi-automated cell selection was integrated. 3D ART VeSElecT removes interfering components, detects vesicles by 3D segmentation and calculates vesicle volume and diameter (spherical approximation, inner/outer diameter). Results are collected in color using the RoiManager plugin including the possibility of manual removal of non-matching confounder vesicles. Detailed evaluation considered performance (detected vesicles) and specificity (true vesicles) as well as precision and recall. We furthermore show gain in segmentation and morphological filtering compared to learning based methods and a large time gain compared to manual segmentation. 3D ART VeSElecT shows small error rates and its speed gain can be up to 68 times faster in comparison to manual annotation. Both automatic and semi-automatic modes are explained including a tutorial.

Awareness and knowledge about HIV mother-to-child transmission (MTCT) and preventive measures in different population groups and health personnel were analysed in future intervention areas in western Uganda and south-western Tanzania. In Uganda, a total of 751 persons (440 clients of antenatal and outpatient clinics, 43 health workers, 239 villagers, 29 traditional birth attendants) and in Tanzania, 574 persons (410 clients, 49 health workers, 93 villagers, 18 traditional birth attendants) were interviewed. When given options, knowledge on transmission during pregnancy and delivery in women was 93% and 67% in Uganda and Tanzania respectively, and 86% and 78% for transmission during breastfeeding. In Uganda 59% of male interviewees did not believe that HIV is transmitted during breastfeeding. Expressed acceptance of HIV testing was above 90% in men and women in both countries, but only 10% of the clients in Uganda and 14% in Tanzania had been tested for HIV infection. Health workers' knowledge regarding MTCT was acceptable, while traditional birth attendants' knowledge on both MTCT and preventive measures was extremely poor. Recommendations on infant feeding were not compatible with WHO recommendations for HIV-infected women. If prevention of MTCT (PMTCT) interventions are to be accepted by the population and promoted by health personnel, thorough orientation and training are mandatory.

The majority of pemphigus vulgaris (PV) patients suffer from a live-threatening loss of intercellular adhesion between keratinocytes (acantholysis). The disease is caused by auto-antibodies that bind to desmosomal cadherins desmoglein (Dsg) 3 or Dsg3 and Dsg1 in mucous membranes and skin. A currently unresolved controversy in PV is whether apoptosis is involved in the pathogenic process. The objective of this study was to perform preclinical studies to investigate apoptotic pathway activation in PV pathogenesis with the goal to assess its potential for clinical therapy. For this purpose, we investigated mouse and human skin keratinocyte cultures treated with PV antibodies (the experimental Dsg3 monospecific antibody AK23 or PV patients IgG), PV mouse models (passive transfer of AK23 or PVIgG into adult and neonatal mice) as well as PV patients' biopsies (n=6). A combination of TUNEL assay, analyses of membrane integrity, early apoptotic markers such as cleaved poly-ADP-ribose polymerase (PARP) and the collapse of actin cytoskeleton failed to provide evidence for apoptosis in PV pathogenesis. However, the in vitro and in vivo PV models, allowing to monitor progression of lesion formation, revealed an early, transient and low-level caspase-3 activation. Pharmacological inhibition confirmed the functional implication of caspase-3 in major events in PV such as shedding of Dsg3, keratin retraction, proliferation including c-Myc induction, p38MAPK activation and acantholysis. Together, these data identify low-level caspase-3 activation downstream of disrupted Dsg3 trans- or cis-adhesion as a major event in PV pathogenesis that is non-synonymous with apoptosis and represents, unlike apoptotic components, a promising target for clinical therapy. At a broader level, these results posit that an impairment of adhesive functions in concert with low-level, non-lethal caspase-3 activation can evoke profound cellular changes which may be of relevance for other diseases including cancer.

Here we report on vaccination approaches against infectious bursal disease (IBD) of poultry that were performed with complete yeast of the species Kluyveromyces lactis (K. lactis). Employing a genetic system that enables the rapid production of stably transfected recombinant K. lactis, we generated yeast strains that expressed defined quantities of the virus capsid forming protein VP2 of infectious bursal disease virus (IBDV). Both, subcutaneous as well as oral vaccination regiments with the heat-inactivated but otherwise untreated yeast induced IBDV-neutralizing antibodies in mice and chickens. A full protection against a subsequent IBDV infection was achieved by subcutaneous inoculation of only milligram amounts of yeast per chicken. Oral vaccination also generated protection: while mortality was observed in control animals after virus challenge, none of the vaccinees died and ca. one-tenth were protected as indicated by the absence of lesions in the bursa of Fabricius. Recombinant K. lactis was thus indicated as a potent tool for the induction of a protective immune response by different applications. Subcutaneously applied K. lactis that expresses the IBDV VP2 was shown to function as an efficacious anti-IBD subunit vaccine.

Random mutagenesis is a useful tool to genetically modify organisms for various purposes, such as adaptation to cultivation conditions, the induction of tolerances, or increased yield of valuable substances. This is especially attractive for systems where it is not obvious which genes require modifications. Random mutagenesis has been extensively used to modify crop plants, but even with the renewed interest in microalgae and cyanobacteria for biofuel applications, there is relatively limited current research available on the application of random mutagenesis for these organisms, especially for cyanobacteria. In the presented work we characterized the lethality and rate of non-lethal point mutations for ultraviolet radiation and methyl methanesulphonate on the model cyanobacteria Synechocystis sp. PCC6803. Based on these results an optimal dosage of 10-50 J/m(2) for UV and either 0.1 or 1 v% for MMS was determined. A Synechocystis wildtype culture was then mutagenized and selected for increased temperature tolerance in vivo. During the second round of mutagenesis the viability of the culture was monitored on a cell by cell level from the treatment of the cells up to the growth at an increased temperature. After four distinct rounds of treatment (two with each mutagen) the temperature tolerance of the strain was effectively raised by about 2°C. Coupled with an appropriate in vivo screening, the described methods should be applicable to induce a variety of desirable characteristics in various strains. Coupling random mutagenesis with high-throughput screening methods would additionally allow to select for important characteristics for biofuel production, which do not yield a higher fitness and can not be selected for in vivo, such as fatty acid concentration. In a combined approach with full genome sequencing random mutagenesis could be used to determine suitable target-genes for more focused methods.