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The successful advancement of xenotransplantation has led to the development of highly sensitive detection systems for the screening of potentially zoonotic viruses in donor pigs and preventing their transmission to the recipient. To validate these methods, genetically modified pigs generated for xenotransplantation, numerous minipigs and other pig breeds have been tested, thereby increasing our knowledge concerning the pig virome and the distribution of pig viruses. Of particular importance are the porcine cytomegalovirus, a porcine roseolovirus (PCMV/PRV) and the hepatitis E virus genotype 3 (HEV3). PCMV/PRV has been shown to reduce the survival time of pig transplants in non-human primates and was also transmitted in the first pig heart transplantation to a human patient. The main aim of this study was to determine the sensitivities of our methods to detect PCMV/PRV, HEV3, porcine lymphotropic herpesvirus-1 (PLHV-1), PLHV-2, PLHV-3, porcine circovirus 2 (PCV2), PCV3, PCV4 and porcine parvovirus 1 (PPV1) and to apply the methods to screen indigenous Greek black pigs. The high number of viruses found in these animals allowed for the evaluation of numerous detection methods. Since porcine endogenous retroviruses (PERVs) type A and B are integrated in the genome of all pigs, but PERV-C is not, the animals were screened for PERV-C and PERV-A/C. Our detection methods were sensitive and detected PCMV/PRV, PLHV-1, PLHV-1, PLHV-3, PVC3 and PERV-C in most animals. PPV1, HEV3, PCV4 and PERV-A/C were not detected. These data are of great interest since the animals are healthy and resistant to diseases.

Xenotransplantation, like allotransplantation, is usually associated with microchimerism, i.e., the presence of cells from the donor in the recipient. Microchimerism was reported in first xenotransplantation trials in humans, as well as in most preclinical trials in nonhuman primates (for review, see Denner, Viruses 2023, 15, 190). When using pigs as xenotransplantation donors, their cells contain porcine endogenous retroviruses (PERVs) in their genome. This makes it difficult to discriminate between microchimerism and PERV infection of the recipient. Here, we demonstrate the appropriate virological methods to be used for the identification of microchimerism, first by screening for porcine cellular genes, and then how to detect infection of the host. Using porcine short interspersed nuclear sequences (SINEs), which have hundreds of thousands of copies in the pig genome, significantly increased the sensitivity of the screening for pig cells. Second, absence of PERV RNA demonstrated an absence of viral genomic RNA or expression as mRNA. Lastly, absence of antibodies against PERV proteins conclusively demonstrated an absence of a PERV infection. When applying these methods for analyzing baboons after pig heart transplantation, microchimerism could be demonstrated and infection excluded in all animals. These methods can be used in future clinical trials.

Erythema multiforme in pigs is an acute, self-limiting disease characterized by red skin areas and often associated with anorexia, fever and respiratory problems. The cause of the disease remains unknown. In a recent study, animals of a commercial breeding herd in Greece were examined, and all animals were found seropositive for porcine reproductive and respiratory syndrome virus (PRRSV). However, neither PRRSV and porcine circovirus type 2 (PCV2) viremia nor antibodies against Aujeszky’s disease virus, African swine fever virus and classical swine fever virus were detected. Here, an extended examination of these pigs was performed on a wide range of porcine viruses using highly sensitive polymerase chain reaction (PCR)-based methods. Affected skin of five animals revealed the presence of porcine lymphotropic herpesvirus-1 (PLHV-1) in all cases, PLHV-2 in one animal and PLHV-3 in four animals. However, neither porcine cytomegalovirus (PCMV) nor porcine circoviruses (PCV1, PCV2, PCV3 and PCV4) were detected. In blood samples, PLHV-1 was present in two animals and PLHV-2, PCV2 and PCV3 in one individual, with PCMV, PCV1 and PCV4 in none of the animals. In one animal, four viruses were found in the blood (PLHV-1, PLHV-2, PCV2 and PCV3). A PRRSV viremia was also not detected. All animals carried porcine endogenous retrovirus C (PERV-C) in their genome, but recombinant PERV-A/C was not detected. The results suggest that porcine viruses may be involved in erythema multiforme in these animals and that further studies are needed to assess the role of these pathogens in the disease.

Minipigs play an important role in biomedical research and have also been used as donor animals in xenotransplantation. To serve as a donor in xenotransplantation, the animals must be free of potential zoonotic viruses, bacteria and parasites. Porcine endogenous retroviruses (PERVs) are integrated in the genome of all pigs and cannot be eliminated as most of the other pig viruses can. PERV-A and PERV-B infect human cells in cell culture and are integrated in all pigs, whereas PERV-C infects only pig cells and it is found in many, but not all pigs. Minipigs are known for a high prevalence of recombinant PERV-A/C viruses able to infect human cells (Denner and Schuurman, Viruses, 2021;13:1869). Here, Mini-LEWE minipigs are screened for the first time for pig viruses including PERV. Peripheral blood mononuclear cells (PBMCs) from 10 animals were screened using PCR-based methods (PCR, RT-PCR, and real-time PCR). In comparison with our previous screening assays, numerous improvements were introduced, e.g., the usage of gene blocks as a PCR standard and foreign RNA to control reverse transcription in RT-PCR. Using these improved detection methods, Mini-LEWE pigs were found to be negative for porcine cytomegalovirus (PCMV), porcine lymphotropic herpesviruses (PLHV-1, -2 and -3), porcine circoviruses (PCV1, 2, 3 and 4), porcine parvovirus (PPV) and hepatitis E virus (HEV). All animals carried PERV-A, PERV-B and PERV-C in their genome. PERV-A/C was not found. In contrast to all other minipig breeds (Göttingen minipigs, Aachen minipigs, Yucatan micropig, Massachusetts General Hospital miniature pigs), Mini-LEWE minipigs have less viruses and no PERV-A/C. Parasitological screening showed that none of the Mini-LEWE minipigs harbored ecto- and gastrointestinal parasites, but at least one animal tested positive for anti-Toxoplasma gondii antibodies.

New town development as a form of large-scale development is not a new phenomenon, particularly in developing countries. This development mainly takes place in peri-urban areas due to the high pressure caused by the growing population and the lack of facilities and infrastructure in city centres. As an effect, local communities who originally occupied the land often lose their rights over the property their livelihood might have relied on. Property rights can be grouped differently, classified according to different bundles: appropriation, ownership, and formality of rights. This paper investigates to what extent new town development in Indonesia has affected the property rights of local communities, in terms of the transformation of rights and security level. Moreover, it examines to what extent this transformation has been affected by urbanisation pressure. Ample attention is paid to the transformation of various bundles of rights concerning different usage of property, both residential and cultivated land. A total of 252 questionnaires were distributed to three different locations of new towns in Indonesia. A before-after analysis was employed to identify the transformation of the property rights and their security level, followed by multiple linear regression analysis to observe the influence of the urbanisation pressure to the security level. The research reveals that the transformation of property rights of local residents mainly concerns the appropriation rights. The analysis also indicates that there is a tendency that the security level decreases. Statistically, this appears to be affected by urbanisation pressure variables: type of land, land use, and occupation. With this study, we offer on the one hand a conceptual framework for assessing property rights, while on the other hand, we provide empirical evidence regarding the effects of new town development on property rights transformation and its security level.

The paper aims to explore the process of land conversion for tourism development in Vietnam, under the present ambiguous and insecure property rights system. Four case studies in different geographical areas were selected to analyse land conversion and land compensation for tourism projects before and after the implementation of the new land law in 2013. The findings of this study show that, in the present legal system of land and property rights, the rights of local people are not sufficiently guaranteed due to the decisive role of the State not only in defining compensation prices for land in the case of compulsory land acquisition but also in determining whether tourism projects are in the public’s interest or not (thus deciding the appropriate land conversion approach as well as affecting price negotiations). The research also found that, although a voluntary land conversion approach (when the project is not in the public’s interest), based on the 2013 Land Law, offers land users a better negotiation position and a higher compensation payment, possibly reducing land-related conflicts between the State and land users, ambiguity over property rights in fact increased due to the government’s substantial discretion to choose between ‘public purpose’ and ‘economic purpose.’ The paper concludes with questioning whether the present legal basis for compulsory land acquisition is future proof since urbanisation pressure is likely to increase, which may lead to even more land conflicts in the near future.

The current paper examines the legitimacy dilemmas that rise from local governments’ direct policy instruments and market interventions. It takes the case of public land management strategies. The paper argues that current societal challenges—such as energy transition, climate change and inclusive urban innovation—require planning practices to be more effective. Direct government instruments such as direct market interventions have proven to significantly reduce the implementation gap of planning practice. Looking at significant urban challenges, municipalities worldwide could be urged to apply such direct government instruments on a larger scale in the future. However, although direct government intervention in markets can be very effective, it is also controversial in terms of legitimacy. It explicitly and inevitably introduces financial incentives to the organization of government. Balancing these incentives against spatial planning interests unavoidably causes dilemmas. Based on eight Dutch case studies, this paper develops a framework to systematically spell out the legitimacy dilemmas that stem from public market intervention. It facilitates an explicit discussion on varying instrumental rationalities and improving the legitimacy of public action.

Ammonia (NH3) emission from agriculture is an environmental threat and a loss of nitrogen for crop production. Mineral fertilizers and manure are significant sources of NH3; therefore, abatement technologies have been introduced to mitigate these emissions. The aim of this study was to demonstrate that low-cost measuring techniques are suitable to assess NH3 emissions in smaller plots, appropriate to test different managements. Two experiments were established to quantify NH3 emissions from urea application in a multi-plot design with radii of 5 (R5) and 20 m (R20). Field was bare soil partially surrounded by shelterbelts. Measurement techniques included passive flux samplers (Leuning), and passive concentration samplers (ALPHA) coupled to WindTrax dispersion model. NH3 emission from R5 was consistent with the emission from R20 when the surface-to-atmosphere exchange was not affected by shelterbelts, and wind speed near surface was greater than 1 m s−1. Both measurement methods gave unreliable NH3 quantification in combination with wind speed lower than 1 m s−1 and low emission strength. Cumulative emission over 60 h was 2% of the supplied N from the plots not affected by the shelterbelt, and 1% from the plots affected by shelterbelts, indicating that these structures can significantly reduce NH3 emissions.