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1.
Sci Rep ; 14(1): 17427, 2024 08 05.
Artículo en Inglés | MEDLINE | ID: mdl-39103377

RESUMEN

The 2018 African swine fever (ASF) outbreak highlighted the importance of biosecurity in food production systems. Despite the significant economic impacts, the sociopsychological consequences on decision-making have been overlooked. Previous studies have focused on algebraic models and simulation-based models without considering the complex psychological and social factors that influence farmers' biosecurity behaviors and decision-making processes. This study aims to classify livestock producers into distinct subgroups based on their attitudes towards biosecurity. We conducted a survey presenting producers with three scenarios to assess their willingness to report suspected ASF cases, trust in government agencies, risk perception, biosecurity knowledge, willingness to purchase livestock insurance, motivation to invest in biosecurity, readiness to report suspected infections, and intention to contact a veterinarian. Using latent class analysis, we identified three distinct classes: Biosecurity Sceptics, Biosecurity Compliant, and Biosecurity Ultra-Compliant. Our results show that producer characteristics significantly influence biosecurity attitudes and class membership, with small-scale producers less likely to adopt ultra-compliant biosecurity practices. Attending at least one eradication program encouraged biosecurity compliance. This research informs the design of targeted food policy and risk communication strategies that account for attitudes of livestock producers to encourage biosecurity adoption and reduce the likelihood of Tier 1 disease incursion.


Asunto(s)
Toma de Decisiones , Agricultores , Animales , Porcinos , Estados Unidos , Agricultores/psicología , Humanos , Bioaseguramiento , Fiebre Porcina Africana/prevención & control , Crianza de Animales Domésticos/métodos , Conocimientos, Actitudes y Práctica en Salud , Encuestas y Cuestionarios , Femenino , Masculino , Actitud
2.
Front Immunol ; 15: 1397780, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39100679

RESUMEN

Enhancing livestock biosecurity is critical to safeguard the livelihoods of farmers, global and local economies, and food security. Vaccination is fundamental to the control and prevention of exotic and endemic high-priority infectious livestock diseases. Successful implementation of vaccination in a biosecurity plan is underpinned by a strong understanding of correlates of protection-those elements of the immune response that can reliably predict the level of protection from viral challenge. While correlates of protection have been successfully characterized for many human viral vaccines, for many high-priority livestock viral diseases, including African swine fever and foot and mouth disease, they remain largely uncharacterized. Current literature provides insights into potential correlates of protection that should be assessed during vaccine development for these high-priority mammalian livestock viral diseases. Establishment of correlates of protection for biosecurity purposes enables immune surveillance, rationale for vaccine development, and successful implementation of livestock vaccines as part of a biosecurity strategy.


Asunto(s)
Ganado , Vacunación , Vacunas Virales , Animales , Ganado/inmunología , Ganado/virología , Vacunas Virales/inmunología , Vacunación/veterinaria , Virosis/prevención & control , Virosis/inmunología , Virosis/veterinaria , Porcinos , Fiebre Aftosa/prevención & control , Fiebre Aftosa/inmunología , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/inmunología , Humanos
3.
J Virol ; 98(7): e0062224, 2024 Jul 23.
Artículo en Inglés | MEDLINE | ID: mdl-38953377

RESUMEN

African swine fever virus causes a lethal hemorrhagic disease in domestic swine and wild boar for which currently licensed commercial vaccines are only available in Vietnam. Development of subunit vaccines is complicated by the lack of information on protective antigens as well as suitable delivery systems. Our previous work showed that a pool of eight African swine fever virus genes vectored using an adenovirus prime and modified vaccinia virus boost could prevent fatal disease after challenge with a virulent genotype I isolate of the virus. Here, we identify antigens within this pool of eight that are essential for the observed protection and demonstrate that adenovirus-prime followed by adenovirus-boost can also induce protective immune responses against genotype I African swine fever virus. Immunization with a pool of adenoviruses expressing individual African swine fever virus genes partially tailored to genotype II virus did not protect against challenge with genotype II Georgia 2007/1 strain, suggesting that different antigens may be required to induce cross-protection for genetically distinct viruses. IMPORTANCE: African swine fever virus causes a lethal hemorrhagic disease in domestic pigs and has killed millions of animals across Europe and Asia since 2007. Development of safe and effective subunit vaccines against African swine fever has been problematic due to the complexity of the virus and a poor understanding of protective immunity. In a previous study, we demonstrated that a complex combination of eight different virus genes delivered using two different viral vector vaccine platforms protected domestic pigs from fatal disease. In this study, we show that three of the eight genes are required for protection and that one viral vector is sufficient, significantly reducing the complexity of the vaccine. Unfortunately, this combination did not protect against the current outbreak strain of African swine fever virus, suggesting that more work to identify immunogenic and protective viral proteins is required to develop a truly effective African swine fever vaccine.


Asunto(s)
Adenoviridae , Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Vectores Genéticos , Genotipo , Vacunas Virales , Animales , Virus de la Fiebre Porcina Africana/genética , Virus de la Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/virología , Fiebre Porcina Africana/inmunología , Porcinos , Vacunas Virales/inmunología , Vacunas Virales/genética , Vacunas Virales/administración & dosificación , Vectores Genéticos/genética , Adenoviridae/genética , Adenoviridae/inmunología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Vacunas de Subunidad/inmunología , Vacunas de Subunidad/genética , Antígenos Virales/inmunología , Antígenos Virales/genética
4.
Prev Vet Med ; 229: 106238, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38870565

RESUMEN

African Swine Fever (ASF) is a contagious viral disease that infects wild and domesticated swine. In early 2022, the virus was found in wild boar in the Apennine mountains of mainland Italy.2 Since then, it has spread from wild boar to domesticated swine. To control the spread of ASF, an effective surveillance system and the implementation of strict biosecurity measures on farms are required yet are unevenly implemented across husbandry systems. Smallholder farms in particular are known to have low levels of biosecurity. In the Apennine mountains of Italy, small commercial farms have been found to have low levels of biosecurity despite being located in areas with high densities of wild boar, and, hence, being high-risk sites for potential ASF incursion and subsequent diffusion. To address the question as to why the level of biosecurity is low, interviews and participant observation were conducted with smallholder commercial farmers. The interviews identified the social, cultural, and ecological factors that affect the implementation of biosecurity measures in small commercial swine farms in the Apennines. Farmers expressed knowledge of priority biosecurity measures and an overall willingness to follow rules and regulations; however, the application of the measures in practice was uneven across farms. Economic, political, and ecological factors as well as farmer beliefs about biosecurity emerged as important factors affecting the implementation of biosecurity measures. These include economic constraints, challenges posed by the mountain environment, a shifting regulatory environment, and ideas about animal welfare. Other important factors include cultural factors such as the use of traditional agricultural methods and norms about customer access to animals, time constraints and the perceived hassle of implementing the measures, farmer age, farmer relationships with government officials and veterinarians, and the role of pigs in reducing farm waste. The study confirmed that wild boar are present in high numbers and in close proximity to smallholder commercial farms in the Apennines.


Asunto(s)
Fiebre Porcina Africana , Crianza de Animales Domésticos , Brotes de Enfermedades , Animales , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/epidemiología , Italia/epidemiología , Porcinos , Crianza de Animales Domésticos/métodos , Brotes de Enfermedades/veterinaria , Brotes de Enfermedades/prevención & control , Sus scrofa , Bioaseguramiento , Agricultores/psicología
6.
Int J Biol Macromol ; 270(Pt 1): 132432, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38761609

RESUMEN

The African swine fever virus (ASFV) continues to pose significant economic and pandemic risks. Consequently, discovering new, efficient vaccines is crucial. Messenger RNA (mRNA) vaccines have emerged as promising candidates, providing minimal risk of insertional mutagenesis, high safety profiles, effectiveness, rapid scalability in production, and cost-effectiveness. In this study, we have developed an ASF p30 mRNA vaccine candidate (mRNA/Man-LNP) employing mannose-modified lipid nanoparticles (LNPs). The mRNA/Man-LNP exhibited effective antigen presentation and facilitated dendritic cells (DCs) maturation. Notably, it elicited strong IgG titers and activated CD4+ and CD8+ T-cells in immunized mice, all while adhering to stringent biosafety standards. This investigation demonstrates that mRNA/Man-LNP can trigger both humoral and cellular immune responses, suggesting its potential as a potent and promising vaccine candidate for controlling African swine fever (ASF).


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Manosa , Nanopartículas , Vacunas Virales , Animales , Nanopartículas/química , Virus de la Fiebre Porcina Africana/inmunología , Virus de la Fiebre Porcina Africana/genética , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/inmunología , Ratones , Vacunas Virales/inmunología , Porcinos , Manosa/química , Células Dendríticas/inmunología , Lípidos/química , Desarrollo de Vacunas , ARN Mensajero/genética , ARN Mensajero/inmunología , Vacunas de ARNm , Femenino , Anticuerpos Antivirales/inmunología , Anticuerpos Antivirales/sangre , Liposomas
7.
Open Vet J ; 14(4): 941-951, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38808296

RESUMEN

African swine fever virus (ASFV) poses a significant threat to global swine populations, necessitating a profound understanding of viral strategies against host antiviral innate immunity. This review synthesizes current knowledge regarding ASFV proteins and their intricate interactions with host defenses. Noteworthy findings encompass the modulation of interferon signaling, manipulation of inflammatory pathways, and the impact on cellular apoptosis. The implications of these findings provide a foundation for advancing vaccine strategies against ASFV. In conclusion, this review consolidates current knowledge, emphasizing the adaptability of ASFV in subverting host immunity. Identified research gaps underscore the need for continued exploration, presenting opportunities for developing targeted vaccines. This synthesis provides a roadmap for future investigations, aiming to enhance our preparedness against the devastating impact of ASFV on global swine populations.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Inmunidad Innata , Proteínas Virales , Vacunas Virales , Virus de la Fiebre Porcina Africana/inmunología , Animales , Porcinos , Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/virología , Proteínas Virales/inmunología , Vacunas Virales/inmunología , Desarrollo de Vacunas
8.
Front Immunol ; 15: 1373656, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38742108

RESUMEN

African swine fever virus (ASFV) is one of the most complex viruses. ASFV is a serious threat to the global swine industry because no commercial vaccines against this virus are currently available except in Vietnam. Moreover, ASFV is highly stable in the environment and can survive in water, feed, and aerosols for a long time. ASFV is transmitted through the digestive and respiratory tract. Mucosal immunity is the first line of defense against ASFV. Saccharomyces cerevisiae (SC), which has been certified by the U.S. Food and Drug Administration and has a generally recognized as safe status in the food industry, was used for oral immunization in this study. ASFV antigens were effectively expressed in recombinant SC strains with high DNA copy numbers and stable growth though surface display technology and chromosome engineering (δ-integration). The recombinant SC strains containing eight ASFV antigens-KP177R, E183L, E199L, CP204L, E248R, EP402R, B602L, and B646L- induced strong humoral and mucosal immune responses in mice. There was no antigenic competition, and these antigens induced Th1 and Th2 cellular immune responses. Therefore, the oral immunization strategy using recombinant SC strains containing multiple ASFV antigens demonstrate potential for future testing in swine, including challenge studies to evaluate its efficacy as a vaccine against ASFV.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Antígenos Virales , Inmunización , Saccharomyces cerevisiae , Vacunas Virales , Animales , Virus de la Fiebre Porcina Africana/inmunología , Virus de la Fiebre Porcina Africana/genética , Saccharomyces cerevisiae/inmunología , Saccharomyces cerevisiae/genética , Administración Oral , Ratones , Vacunas Virales/inmunología , Vacunas Virales/administración & dosificación , Antígenos Virales/inmunología , Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/prevención & control , Porcinos , Inmunidad Mucosa , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Ratones Endogámicos BALB C , Femenino , Inmunidad Humoral
9.
Microbiol Spectr ; 12(7): e0216423, 2024 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-38563791

RESUMEN

African swine fever (ASF) is a highly fatal viral disease that poses a significant threat to domestic pigs and wild boars globally. In our study, we aimed to explore the potential of a multiplexed CRISPR-Cas system in suppressing ASFV replication and infection. By engineering CRISPR-Cas systems to target nine specific loci within the ASFV genome, we observed a substantial reduction in viral replication in vitro. This reduction was achieved through the concerted action of both Type II and Type III RNA polymerase-guided gRNA expression. To further evaluate its anti-viral function in vivo, we developed a pig strain expressing the multiplexable CRISPR-Cas-gRNA via germline genome editing. These transgenic pigs exhibited normal health with continuous expression of the CRISPR-Cas-gRNA system, and a subset displayed latent viral replication and delayed infection. However, the CRISPR-Cas9-engineered pigs did not exhibit a survival advantage upon exposure to ASFV. To our knowledge, this study represents the first instance of a living organism engineered via germline editing to assess resistance to ASFV infection using a CRISPR-Cas system. Our findings contribute valuable insights to guide the future design of enhanced viral immunity strategies. IMPORTANCE: ASFV is currently a devastating disease with no effective vaccine or treatment available. Our study introduces a multiplexed CRISPR-Cas system targeting nine specific loci in the ASFV genome. This innovative approach successfully inhibits ASFV replication in vitro, and we have successfully engineered pig strains to express this anti-ASFV CRISPR-Cas system constitutively. Despite not observing survival advantages in these transgenic pigs upon ASFV challenges, we did note a delay in infection in some cases. To the best of our knowledge, this study constitutes the first example of a germline-edited animal with an anti-virus CRISPR-Cas system. These findings contribute to the advancement of future anti-viral strategies and the optimization of viral immunity technologies.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Sistemas CRISPR-Cas , Edición Génica , Replicación Viral , Animales , Virus de la Fiebre Porcina Africana/genética , Porcinos , Fiebre Porcina Africana/virología , Fiebre Porcina Africana/inmunología , Fiebre Porcina Africana/prevención & control , Edición Génica/métodos , Replicación Viral/genética , Animales Modificados Genéticamente/genética , ARN Guía de Sistemas CRISPR-Cas/genética , Genoma Viral/genética
10.
Prev Vet Med ; 227: 106193, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38626594

RESUMEN

Animal disease outbreaks, such as the recent outbreak of African Swine Fever in 2018, are a major concern for stakeholders across the food supply chain due to their potential to disrupt global food security, cause economic losses, and threaten animal welfare. As a result of their transboundary nature, discussions have shifted to preventive measures aimed at protecting livestock while ensuring food security and safety. Emergency assistance has been a critical response option during pandemics. However, this may not be sustainable in the long run because the expectation of government bailout may encourage risk taking behaviours. Our hypothesis is that an indemnity policy that is conditioned on showing biosecurity practices would increase compliance and reduce government expenditure during disease outbreaks. We developed and launched a survey from March to July 2022 targeted at swine producers across the US. From the survey, we examined livestock farmers' attitudes and intentions regarding biosecurity investment and assessed their attitudes towards the purchase of livestock insurance and reporting suspected infected livestock on their farm. We used a partial proportion odds model analysis to examine the model. Our analysis revealed that intention to call a veterinarian, trust in government agencies and risk perception of farmers were instrumental in the willingness to self-invest in biosecurity, purchase livestock insurance, and promptly report infected livestock on their farms. This provides evidence that biosecurity compliance would increase if indemnification was tied to a demonstration of effort to adopt biosecurity practices. We also show that individuals who have been in the industry for a longer period may become complacent and less likely to report outbreaks. Farmers with a higher share of income from their production operations bear a greater risk from their operational income and are more willing to report any suspected infections on their farms. The data suggest that motivating the willingness of farmers to invest in biosecurity while overcoming cost concerns is achievable.


Asunto(s)
Fiebre Porcina Africana , Brotes de Enfermedades , Agricultores , Animales , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/epidemiología , Fiebre Porcina Africana/psicología , Estados Unidos/epidemiología , Brotes de Enfermedades/veterinaria , Brotes de Enfermedades/prevención & control , Porcinos , Agricultores/psicología , Crianza de Animales Domésticos/métodos , Bioaseguramiento , Humanos , Conocimientos, Actitudes y Práctica en Salud , Masculino , Femenino , Encuestas y Cuestionarios
11.
Vet Q ; 44(1): 1-13, 2024 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38533618

RESUMEN

Despite the great interest in the development of a vaccine against African swine fever (ASF) in wild boar, the immunological mechanisms that induce animal protection are still unknown. For this purpose, tertiary lymphoid organs (TLOs) of wild boar were characterised and compared with mucosa-associated lymphoid tissues (MALTs) by histopathology, histomorphometry and immunohistochemistry (CD3, CD79, PAX5, LYVE1, fibronectin). In addition, real-time polymerase chain reaction (qPCR) and immunohistochemistry (p72) were used to evaluate the presence of ASF virus (ASFV) in blood and tissues samples, respectively. TLOs were observed in animals infected with a low-virulent ASFV isolate (LVI), animals co-infected with low and high-virulent ASFV isolates (LVI-HVI) and animals infected only with the high virulence isolate (HVI). TLOs in LVI and LVI-HVI groups were located adjacent to the mucosa and presented a similar structure to MALT. Immunoexpresion of p72 observed in the inflammatory cells adjacent to TLOs/MALTs confirmed its development and reactivity generated by ASF attenuated isolates. Immunohistochemical evaluation, based on cellular composition (T and B lymphocytes), and histomorphometrical study revealed a more pronounced maturation of TLOs/MALTs in the LVI-HVI group. It is currently unclear whether these formations play a protective role by contributing to local immunity in chronic inflammatory diseases. However, the structural similarities between TLOs and MALTs and the location of TLOs close to the mucosa suggest that they may perform a similar function, facilitating a local protective response. Nevertheless, further investigations are warranted to assess the cellular and humoral dynamics of these lymphoid organs induced by attenuated isolates.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Enfermedades de los Porcinos , Porcinos , Animales , Sus scrofa , Virus de la Fiebre Porcina Africana/fisiología , Fiebre Porcina Africana/prevención & control , Virulencia
12.
Prev Vet Med ; 226: 106168, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38507888

RESUMEN

Several propagation routes drive animal disease dissemination, and among these routes, contaminated vehicles traveling between farms have been associated with indirect disease transmission. In this study, we used near-real-time vehicle movement data and vehicle cleaning efficacy to reconstruct the between-farm dissemination of the African swine fever virus (ASFV). We collected one year of Global Positioning System data of 823 vehicles transporting feed, pigs, and people to 6363 swine production farms in two regions in the U.S. Without cleaning, vehicles connected up to 2157 farms in region one and 437 farms in region two. Individually, in region one vehicles transporting feed connected 2151 farms, pigs to farms 2089 farms, pigs to market 1507 farms, undefined vehicles 1760 farm, and personnel three farms. The simulation results indicated that the contact networks were reduced the most for crew transport vehicles with a 66% reduction, followed by vehicles carrying pigs to market and farms, with reductions of 43% and 26%, respectively, when 100% cleaning efficacy was achieved. The results of this study showed that even when vehicle cleaning and disinfection are 100% effective, vehicles are still connected to numerous farms. This emphasizes the importance of better understanding transmission risks posed by vehicles to the swine industry and regulatory agencies.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Enfermedades de los Porcinos , Humanos , Porcinos , Animales , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/prevención & control , Granjas , Incertidumbre , Simulación por Computador , Fiebre Porcina Africana/epidemiología , Fiebre Porcina Africana/prevención & control , Brotes de Enfermedades/veterinaria
13.
Int J Biol Macromol ; 264(Pt 2): 130689, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38458287

RESUMEN

African Swine Fever Virus (ASFV) is a highly contagious pathogen posing a serious threat to the global swine industry. Despite this, there is currently no effective vaccine against this virus. Within ASFV's core shell structure, p37, a product of polyprotein pp220, shares sequence similarity with SUMO-1 proteases. Localization studies show p37 in various nuclear regions during early infection, shifting to the cytoplasm later on. Research indicates active export of p37 from the nucleus, mediated by CRM1-dependent and -independent pathways. Hydrophobic amino acids in p37 are crucial for these pathways, highlighting their importance throughout the ASFV replication cycle. Additionally, p37 serves as the first nucleocytoplasmic shuttle protein encoded by ASFV, participating in the intranuclear material transport process during ASFV infection of host cells. In this study, we successfully screened five murine monoclonal antibodies targeting p37. Through the truncated expression method, we identified four dominant antigenic epitopes of p37 for the first time. Furthermore, utilizing alanine scanning technology, we determined the key amino acid residues for each epitope. This research not only provides essential information for a deeper understanding of the protein's function but also establishes a significant theoretical foundation for the design and development of ASFV vaccines.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Porcinos , Animales , Ratones , Anticuerpos Monoclonales , Proteínas Virales/química , Fiebre Porcina Africana/prevención & control
14.
Viruses ; 16(3)2024 02 22.
Artículo en Inglés | MEDLINE | ID: mdl-38543702

RESUMEN

In the event of an outbreak of African swine fever (ASF) in pig farms, the European Union (EU) legislation requires the establishment of a restricted zone, consisting of a protection zone with a radius of at least 3 km and a surveillance zone with a radius of at least 10 km around the outbreak. The main purpose of the restricted zone is to stop the spread of the disease by detecting further outbreaks. We evaluated the effectiveness and necessity of the restricted zone in the Baltic States by looking at how many secondary outbreaks were detected inside and outside the protection and surveillance zones and by what means. Secondary outbreaks are outbreaks with an epidemiological link to a primary outbreak while a primary outbreak is an outbreak that is not epidemiologically linked to any previous outbreak. From 2014 to 2023, a total of 272 outbreaks in domestic pigs were confirmed, where 263 (96.7%) were primary outbreaks and 9 (3.3%) were secondary outbreaks. Eight of the secondary outbreaks were detected by epidemiological enquiry and one by passive surveillance. Epidemiological enquiries are legally required investigations on an outbreak farm to find out when and how the virus entered the farm and to obtain information on contact farms where the ASF virus may have been spread. Of the eight secondary outbreaks detected by epidemiological investigations, six were within the protection zone, one was within the surveillance zone and one outside the restricted zone. Epidemiological investigations were therefore the most effective means of detecting secondary outbreaks, whether inside or outside the restricted zones, while active surveillance was not effective. Active surveillance are legally prescribed activities carried out by the competent authorities in the restricted zones. Furthermore, as ASF is no longer a rare and exotic disease in the EU, it could be listed as a "Category B" disease, which in turn would allow for more flexibility and "tailor-made" control measures, e.g., regarding the size of the restricted zone.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Porcinos , Animales , Fiebre Porcina Africana/epidemiología , Fiebre Porcina Africana/prevención & control , Sus scrofa , Brotes de Enfermedades/prevención & control , Brotes de Enfermedades/veterinaria , Países Bálticos
15.
J Microbiol ; 62(2): 125-134, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38480615

RESUMEN

African swine fever virus (ASFV) is the causative agent of the highly lethal African swine fever disease that affects domestic pigs and wild boars. In spite of the rapid spread of the virus worldwide, there is no licensed vaccine available. The lack of a suitable cell line for ASFV propagation hinders the development of a safe and effective vaccine. For ASFV propagation, primary swine macrophages and monocytes have been widely studied. However, obtaining these cells can be time-consuming and expensive, making them unsuitable for mass vaccine production. The goal of this study was to validate the suitability of novel CA-CAS-01-A (CAS-01) cells, which was identified as a highly permissive cell clone for ASFV replication in the MA-104 parental cell line for live attenuated vaccine development. Through a screening experiment, maximum ASFV replication was observed in the CAS-01 cell compared to other sub-clones of MA-104 with 14.89 and log10 7.5 ± 0.15 Ct value and TCID50/ml value respectively. When CAS-01 cells are inoculated with ASFV, replication of ASFV was confirmed by Ct value for ASFV DNA, HAD50/ml assay, TCID50/ml assay, and cytopathic effects and hemadsoption were observed similar to those in primary porcine alveolar macrophages after 5th passage. Additionally, we demonstrated stable replication and adaptation of ASFV over the serial passage. These results suggest that CAS-01 cells will be a valuable and promising cell line for ASFV isolation, replication, and development of live attenuated vaccines.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Porcinos , Animales , Virus de la Fiebre Porcina Africana/genética , Fiebre Porcina Africana/prevención & control , Vacunas Atenuadas/genética , Proteínas Virales/genética , Sus scrofa , Desarrollo de Vacunas , Línea Celular
16.
Nat Commun ; 15(1): 1606, 2024 Feb 21.
Artículo en Inglés | MEDLINE | ID: mdl-38383525

RESUMEN

African Swine Fever Virus is a Nucleo-Cytoplasmic Large DNA Virus that causes an incurable haemorrhagic fever in pigs with a high impact on global food security. ASFV replicates in the cytoplasm of the infected cell and encodes its own transcription machinery that is independent of cellular factors, however, not much is known about how this system works at a molecular level. Here, we present methods to produce recombinant ASFV RNA polymerase, functional assays to screen for inhibitors, and high-resolution cryo-electron microscopy structures of the ASFV RNAP in different conformational states. The ASFV RNAP bears a striking resemblance to RNAPII with bona fide homologues of nine of its twelve subunits. Key differences include the fusion of the ASFV assembly platform subunits RPB3 and RPB11, and an unusual C-terminal domain of the stalk subunit vRPB7 that is related to the eukaryotic mRNA cap 2´-O-methyltransferase 1. Despite the high degree of structural conservation with cellular RNA polymerases, the ASFV RNAP is resistant to the inhibitors rifampicin and alpha-amanitin. The cryo-EM structures and fully recombinant RNAP system together provide an important tool for the design, development, and screening of antiviral drugs in a low biosafety containment environment.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , ARN , Porcinos , Animales , Virus de la Fiebre Porcina Africana/genética , Fiebre Porcina Africana/genética , Fiebre Porcina Africana/prevención & control , Microscopía por Crioelectrón , ARN Polimerasas Dirigidas por ADN/genética , Sus scrofa
17.
Microbiol Spectr ; 12(3): e0140123, 2024 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-38305163

RESUMEN

The African swine fever virus (ASFV) structural protein pA104R is the only histone-like protein encoded by eukaryotic viruses. pA104R is an essential DNA-binding protein required for DNA replication and genome packaging of ASFV, which are vital for pathogen survival and proliferation. pA104R is an important target molecule for diagnosing, treating, and immune prevention of ASFV. This study characterized monoclonal antibodies (mAbs) against pA104R and found them to recognize natural pA104R in ASFV strains with different genotypes, showing high conservation. Confirmation analyses of pA104R epitopes using mAbs indicated the presence of immunodominant B-cell epitopes, and further characterization showed the high antigenic index and surface accessibility coefficients of the identified epitope. Furthermore, the pA104R protein functions through the polar interactions between the binding amino acid sites; however, these interactions may be blocked by the recognition of generated mAbs. Characterizing the immunodominant B-cell epitope of the ASFV critical proteins, such as pA104R, may contribute to developing sensitive diagnostic tools and vaccine candidate targets.IMPORTANCEAfrican swine fever (ASF) is a highly pathogenic, lethal, and contagious viral disease affecting domestic pigs and wild boars. As no effective vaccine or other treatments have been developed, the control of African swine fever virus (ASFV) relies heavily on virus detection and diagnosis. A potential serological target is the structural protein pA104R. However, the molecular basis of pA104R antigenicity remains unclear, and a specific monoclonal antibody (mAb) against this protein is still unavailable. In this study, mAbs against pA104R were characterized and found to recognize natural pA104R in ASFV strains with different genotypes. In addition, confirmation analyses of pA104R epitopes using mAbs indicated the presence of immunodominant B-cell epitopes, and further characterization showed the high antigenic index and surface accessibility coefficients of the identified epitope. Characteristics of the immunodominant B-cell epitope of ASFV proteins, such as pA104R, may contribute to developing sensitive diagnostic tools and identifying vaccine candidate targets.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Vacunas , Ratones , Porcinos , Animales , Virus de la Fiebre Porcina Africana/genética , Epítopos de Linfocito B , Fiebre Porcina Africana/diagnóstico , Fiebre Porcina Africana/prevención & control , Anticuerpos Monoclonales , Sus scrofa
18.
ACS Appl Bio Mater ; 7(3): 1547-1557, 2024 03 18.
Artículo en Inglés | MEDLINE | ID: mdl-38346262

RESUMEN

African swine fever is an acute and highly contagious infectious disease with a mortality rate of up to 100%. The lack of commercial vaccines and drugs is a serious economic threat to the global pig industry. Cell-mediated immunity plays an essential role in protection against viral infection. We previously reported the rational design of a T-cell-activating thermostable scaffold (RPT) for antigen delivery and improved cellular immunity. We conjugated antigens P30, P54, P72, CD2 V, and CP312R to RPT, using a SpyCatcher/SpyTag covalent attachment strategy to construct nanovaccines (multiantigens-RPT). Multiantigens-RPT exhibited significantly higher thermal, storage, and freeze-thaw stability. The specific antibodies IgG and IgG2a of the multiantigen-RPT-immunized were higher than the antigens cocktail-immunized by approximately 10-100 times. ELISpot demonstrated that more IFN-γ-secreting cells were produced by the multiantigen-RPT-immunized than by the antigens cocktail-immunized. Delivery of the multiantigen nanovaccine by a T-cell-activating scaffold induced strong humoral and cellular immune responses in mice and pigs and is a potentially useful candidate vaccine for the African swine fever virus.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Porcinos , Animales , Ratones , Fiebre Porcina Africana/prevención & control , Linfocitos T , Nanovacunas , Adyuvantes Inmunológicos
19.
Braz J Microbiol ; 55(1): 997-1010, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38311710

RESUMEN

The swine industry across the globe is recently facing a devastating situation imparted by a highly contagious and deadly viral disease, African swine fever. The disease is caused by a DNA virus, the African swine fever virus (ASFV) of the genus Asfivirus. ASFV affects both wild boars and domestic pigs resulting in an acute form of hemorrhagic fever. Since the first report in 1921, the disease remains endemic in some of the African countries. However, the recent occurrence of ASF outbreaks in Asia led to a fresh and formidable challenge to the global swine production industry. Culling of the infected animals along with the implementation of strict sanitary measures remains the only options to control this devastating disease. Efforts to develop an effective and safe vaccine against ASF began as early as in the mid-1960s. Different approaches have been employed for the development of effective ASF vaccines including inactivated vaccines, subunit vaccines, DNA vaccines, virus-vectored vaccines, and live attenuated vaccines (LAVs). Inactivated vaccines are a non-feasible strategy against ASF due to their inability to generate a complete cellular immune response. However genetically engineered vaccines, such as subunit vaccines, DNA vaccines, and virus vector vaccines, represent tailored approaches with minimal adverse effects and enhanced safety profiles. As per the available data, gene deleted LAVs appear to be the most potential vaccine candidates. Currently, a gene deleted LAV (ASFV-G-∆I177L), developed in Vietnam, stands as the sole commercially available vaccine against ASF. The major barrier to the goal of developing an effective vaccine is the critical gaps in the knowledge of ASFV biology and the immune response induced by ASFV infection. The precise contribution of various hosts, vectors, and environmental factors in the virus transmission must also be investigated in depth to unravel the disease epidemiology. In this review, we mainly focus on the recent progress in vaccine development against ASF and the major gaps associated with it.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Vacunas de ADN , Vacunas Virales , Porcinos , Animales , Fiebre Porcina Africana/prevención & control , Fiebre Porcina Africana/epidemiología , Virus de la Fiebre Porcina Africana/genética , Vacunas de ADN/genética , Sus scrofa , Vacunas Virales/genética , Vacunas Atenuadas/genética , Desarrollo de Vacunas , Vacunas de Productos Inactivados , Vacunas de Subunidad
20.
Virology ; 593: 110014, 2024 05.
Artículo en Inglés | MEDLINE | ID: mdl-38401340

RESUMEN

African swine fever (ASF) caused by African swine fever virus (ASFV) is a highly infectious and lethal swine disease. Currently, there is only one novel approved vaccine and no antiviral drugs for ASFV. In the study, a high-throughput screening of an FDA-approved drug library was performed to identify several drugs against ASFV infection in primary porcine alveolar macrophages. Triapine and cytarabine hydrochloride were identified as ASFV infection inhibitors in a dose-dependent manner. The two drugs executed their antiviral activity during the replication stage of ASFV. Furthermore, molecular docking studies showed that triapine might interact with the active center Fe2+ in the small subunit of ASFV ribonucleotide reductase while cytarabine hydrochloride metabolite might interact with three residues (Arg589, Lys593, and Lys631) of ASFV DNA polymerase to block new DNA chain extension. Taken together, our results suggest that triapine and cytarabine hydrochloride displayed significant antiviral activity against ASFV in vitro.


Asunto(s)
Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Piridinas , Tiosemicarbazonas , Porcinos , Animales , Virus de la Fiebre Porcina Africana/genética , Virus de la Fiebre Porcina Africana/metabolismo , Fiebre Porcina Africana/prevención & control , Simulación del Acoplamiento Molecular , Antivirales/farmacología , Antivirales/metabolismo , Citarabina/metabolismo , Citarabina/farmacología , Replicación Viral
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