The African Swine Fever Isolate ASFV-Kenya-1033-IX is highly virulent and stable after growth in the wild boar cell line WSL

2021 ◽  
Author(s):  
Johanneke D. Hemmink ◽  
Hussein M. Abkallo ◽  
Sonal P. Henson ◽  
Emmanuel M. Khazalwa ◽  
Bernard Oduor ◽  
...  
Keyword(s):  
Wild Boar ◽  
Cell Line ◽  
Genome Stability ◽  
Central Africa ◽  
African Swine Fever ◽  
Single Nucleotide ◽  
In The Wild ◽  
Genotype Ix ◽  
Set Up

In this study, we describe an African swine fever genotype IX virus (ASFV-Kenya-1033-IX), which was isolated from a domestic pig in Western Kenya during a reported outbreak, including efficiency of virus replication, in vivo virulence, and genome stability in pulmonary alveolar macrophages (PAM) and in a wild boar cell line (WSL). The ASFV-Kenya-1033-IX stock, which underwent multiple passages in WSL (more than 20), retained its ability to replicate in primary macrophages and it also retained the virulence in vivo. At the genomic level, only a few single nucleotide differences were observed between the macrophage and WSL-grown virus. Thus, we propose that the WSL cell line is suitable to produce live attenuated ASFV vaccine candidates based on this isolate and probably of similar viruses. The genome sequences for ASFV-Kenya-1033-IX grown in macrophages and in WSL cells was submitted to GenBank and a challenge model based on this isolate was set up, which will aid the development of vaccines against genotype IX ASFV circulating in Eastern and Central Africa.

scholarly journals Involvement of the Reparative DNA Polymerase Pol X of African Swine Fever Virus in the Maintenance of Viral Genome Stability In Vivo

2013 ◽  
Vol 87 (17) ◽  
pp. 9780-9787 ◽  
Author(s):  
M. Redrejo-Rodriguez ◽  
J. M. Rodriguez ◽  
C. Suarez ◽  
J. Salas ◽  
M. L. Salas
Keyword(s):  
Dna Polymerase ◽  
Viral Genome ◽  
Genome Stability ◽  
African Swine Fever Virus ◽  
African Swine Fever ◽  
Fever Virus

scholarly journals Thermodynamically stable and genetically unstable G-quadruplexes are depleted in genomes across species

2019 ◽  
Vol 47 (12) ◽  
pp. 6098-6113 ◽  
Author(s):  
Emilia Puig Lombardi ◽  
Allyson Holmes ◽  
Daniela Verga ◽  
Marie-Paule Teulade-Fichou ◽  
Alain Nicolas ◽  
...  
Keyword(s):  
Negative Selection ◽  
Genome Stability ◽  
Genome Instability ◽  
Regulation Of Gene Expression ◽  
Biological Processes ◽  
Single Nucleotide ◽  
Folding Potential ◽  
Species Specific

Abstract G-quadruplexes play various roles in multiple biological processes, which can be positive when a G4 is involved in the regulation of gene expression or detrimental when the folding of a stable G4 impairs DNA replication promoting genome instability. This duality interrogates the significance of their presence within genomes. To address the potential biased evolution of G4 motifs, we analyzed their occurrence, features and polymorphisms in a large spectrum of species. We found extreme bias of the short-looped G4 motifs, which are the most thermodynamically stable in vitro and thus carry the highest folding potential in vivo. In the human genome, there is an over-representation of single-nucleotide-loop G4 motifs (G4-L1), which are highly conserved among humans and show a striking excess of the thermodynamically least stable G4-L1A (G3AG3AG3AG3) sequences. Functional assays in yeast showed that G4-L1A caused the lowest levels of both spontaneous and G4-ligand-induced instability. Analyses across 600 species revealed the depletion of the most stable G4-L1C/T quadruplexes in most genomes in favor of G4-L1A in vertebrates or G4-L1G in other eukaryotes. We discuss how these trends might be the result of species-specific mutagenic processes associated to a negative selection against the most stable motifs, thus neutralizing their detrimental effects on genome stability while preserving positive G4-associated biological roles.

scholarly journals Rhes, a striatal-enriched protein, promotes mitophagy via Nix

2019 ◽  
Vol 116 (47) ◽  
pp. 23760-23771 ◽  
Author(s):  
Manish Sharma ◽  
Uri Nimrod Ramírez-Jarquín ◽  
Oscar Rivera ◽  
Melissa Kazantzis ◽  
Mehdi Eshraghi ◽  
...  
Keyword(s):  
Cell Death ◽  
Cell Line ◽  
Neuronal Cell ◽  
Dependent Manner ◽  
Systemic Injection ◽  
Neuronal Cell Line ◽  
In The Wild ◽  
Striatal Lesion ◽  
The Brain

Elimination of dysfunctional mitochondria via mitophagy is essential for cell survival and neuronal functions. But, how impaired mitophagy participates in tissue-specific vulnerability in the brain remains unclear. Here, we find that striatal-enriched protein, Rhes, is a critical regulator of mitophagy and striatal vulnerability in brain. In vivo interactome and density fractionation reveal that Rhes coimmunoprecipitates and cosediments with mitochondrial and lysosomal proteins. Live-cell imaging of cultured striatal neuronal cell line shows Rhes surrounds globular mitochondria, recruits lysosomes, and ultimately degrades mitochondria. In the presence of 3-nitropropionic acid (3-NP), an inhibitor of succinate dehydrogenase, Rhes disrupts mitochondrial membrane potential (ΔΨm) and promotes excessive mitophagy and cell death. Ultrastructural analysis reveals that systemic injection of 3-NP in mice promotes globular mitochondria, accumulation of mitophagosomes, and striatal lesion only in the wild-type (WT), but not in the Rhes knockout (KO), striatum, suggesting that Rhes is critical for mitophagy and neuronal death in vivo. Mechanistically, Rhes requires Nix (BNIP3L), a known receptor of mitophagy, to disrupt ΔΨm and promote mitophagy and cell death. Rhes interacts with Nix via SUMO E3-ligase domain, and Nix depletion totally abrogates Rhes-mediated mitophagy and cell death in the cultured striatal neuronal cell line. Finally, we find that Rhes, which travels from cell to cell via tunneling nanotube (TNT)-like cellular protrusions, interacts with dysfunctional mitochondria in the neighboring cell in a Nix-dependent manner. Collectively, Rhes is a major regulator of mitophagy via Nix, which may determine striatal vulnerability in the brain.

scholarly journals African Swine Fever Status in Europe

Viruses ◽  
2019 ◽  
Vol 11 (4) ◽  
pp. 310 ◽  
Author(s):  
Przemyslaw Cwynar ◽  
Jane Stojkov ◽  
Klaudia Wlazlak
Keyword(s):  
Eastern Europe ◽  
Wild Boar ◽  
Western Europe ◽  
African Swine Fever ◽  
Disease Spread ◽  
Wild Boar Population ◽  
Domestic Pig ◽  
Domestic Pigs ◽  
In The Wild ◽  
Virus Spreading

African Swine Fever (ASF) is a highly contagious disease that affects the domestic pig and wild boar population. The aim of this study was to describe the introduction and spread of the ASF virus in Western Europe (1960–1995) and in Eastern Europe (2007–2018), with particular emphasis on the current ASF situation in Poland and its challenges and future perspectives. The first ASF outbreak in Europe was reported in Portugal in 1957, with the virus spreading over most of Western Europe over the next 30 years. In Eastern Europe, the virus was first observed in Georgia in 2007, from where the disease spread quickly to other neighboring countries, reaching Poland in 2014. Since then, there have been 3341 confirmed cases in the wild boar population in Poland. Although there have been no confirmed cases of wild boars coming into contact with domestic pigs, the first notified case concerning domestic pigs was reported in July 2014. Since then, there have been a total of 213 confirmed outbreaks of ASF on Polish pig farms. Given the virulence of the ASF virus and the myriad of transmission routes across Europe, the monitoring of this disease must be a priority for Europe.

scholarly journals African Swine Fever in Wild Boar (Poland 2020): Passive and Active Surveillance Analysis and Further Perspectives

Pathogens ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1219
Author(s):  
Maciej Piotr Frant ◽  
Anna Gal-Cisoń ◽  
Łukasz Bocian ◽  
Anna Ziętek-Barszcz ◽  
Krzysztof Niemczuk ◽  
...  
Keyword(s):  
Wild Boar ◽  
Western Europe ◽  
African Swine Fever ◽  
Hemorrhagic Disease ◽  
Wild Boar Population ◽  
Domestic Pigs ◽  
In The Wild ◽  
Surveillance Analysis ◽  
Eastern And Western Europe

African swine fever (ASF) is a fatal hemorrhagic disease of wild boar and domestic pigs which has been present in Poland since 2014. By 2020, the ASF virus (ASFV) spread across Central, Eastern and Western Europe (including Germany), and Asian countries (including China, Vietnam, and South Korea). The national ASF eradication and prevention program includes continuous passive (wild boar found dead and road-killed wild boar) and active (hunted wild boar) surveillance. The main goal of this study was to analyze the dynamic of the spread of ASF in the wild boar population across the territory of Poland in 2020. In that year in Poland, in total 6191 ASF-positive wild boar were declared. Most of them were confirmed in a group of animals found dead. The conducted statistical analysis indicates that the highest chance of obtaining an ASF-positive result in wild boar was during the winter months, from January to March, and in December 2020. Despite the biosecurity measures implemented by holdings of domestic pigs, the disease also occurred in 109 pig farms. The role of ASF surveillance in the wild boar population is crucial to apply more effective and tailored measures of disease control and eradication. The most essential measures to maintain sustainable production of domestic pigs in Poland include effective management of the wild boar population, along with strict implementation of biosecurity measures by domestic pig producers.

scholarly journals Stability of Murine Cytomegalovirus Genome after In Vitro and In Vivo Passage

2009 ◽  
Vol 84 (5) ◽  
pp. 2623-2628 ◽  
Author(s):  
Tammy P. Cheng ◽  
Mark C. Valentine ◽  
Jian Gao ◽  
Jeanette T. Pingel ◽  
Wayne M. Yokoyama
Keyword(s):  
Selection Pressure ◽  
Genome Stability ◽  
Murine Cytomegalovirus ◽  
Mutation Rates ◽  
Nucleotide Level ◽  
Single Nucleotide ◽  
Dna Viruses ◽  
Single Base Pair

ABSTRACT While large DNA viruses are thought to have low mutation rates, only a small fraction of their genomes have been analyzed at the single-nucleotide level. Here, we defined the genetic stability of murine cytomegalovirus (MCMV) by whole-genome sequencing. Independently assembled sequences of three sister plaques showed only two single-base-pair substitutions after in vitro passage. In vivo-passaged MCMV likewise demonstrated low mutation rates, comparable to those after in vitro passage, indicating high genome stability of MCMV at the single-nucleotide level in the absence of obvious selection pressure.

Monitoring of African Swine Fever in the Wild Boar Population of the Most Recent Endemic Area of Spain

2012 ◽  
Vol 59 (6) ◽  
pp. 526-531 ◽  
Author(s):  
L. Mur ◽  
M. Boadella ◽  
B. Martínez-López ◽  
C. Gallardo ◽  
C. Gortazar ◽  
...  
Keyword(s):  
Wild Boar ◽  
Endemic Area ◽  
African Swine Fever ◽  
Wild Boar Population ◽  
In The Wild

scholarly journals The Spillover of African Swine Fever in Western Poland Revealed Its Estimated Origin on the Basis of O174L, K145R, MGF 505-5R and IGR I73R/I329L Genomic Sequences

Viruses ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 1094 ◽  
Author(s):  
Natalia Mazur-Panasiuk ◽  
Marek Walczak ◽  
Małgorzata Juszkiewicz ◽  
Grzegorz Woźniakowski
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Wild Boar ◽  
Tandem Repeats ◽  
African Swine Fever ◽  
Whole Genome Sequence ◽  
Reference Sequence ◽  
Nucleotide Sequencing ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
First Case

The African swine fever epidemic occurred in Poland at the beginning of 2014 and, up to date, the disease has been spreading mainly in the eastern part of the country. Unexpectedly, in November 2019 an infected wild boar case was confirmed in Lubuskie voivodship in western Poland. During the following weeks, several dozen African swine fever virus (ASFV)-positive animals were notified in the neighboring area, causing severe concern regarding further spread of the disease to the mostly pig-dense region in Poland, namely, Wielkopolskie voivodship. Moreover, almost a year after, several infected wild boar cases were confirmed for the first time in Germany, just beyond the Polish border, sending out a shock wave through the global pig market. The whole genome sequence of ASFV, isolated from the first case of ASF in western Poland, and three selected viruses from other affected areas, revealed the tandem repeat and single nucleotide polymorphism (SNP) variations in reference to the Georgia 2007/1 strain. These data, supported by the conventional sequencing of selected genomic regions from a total of 154 virus samples isolated between 2017 and 2020 in Poland, shed a new light on pathogen epidemiology. The sequence variations within the O174L gene detected in this study showed that cases identified in western Poland might be originating from the so-called southern Warsaw cluster. Moreover, the viruses originating from the northern Warsaw cluster do not possess single nucleotide polymorphism (SNP) mutations within the K145R and MGF 505-5R genes, which are specific to all of the other Polish ASFV strains. These results led to a conclusion of their distinct origin. Supporting these results, the nucleotide sequencing of I73R/I329L intergenic region revealed its new, previously undescribed variant, called IGR IV, with an additional three tandem repeats of 10 nucleotides in comparison to the reference sequence of the Georgia 2007/1 strain.

scholarly journals African swine fever virus (ASFV) in Poland: Prevalence in a wild boar population (2017–2018)

2020 ◽  
Vol 65 (No. 4) ◽  
pp. 143-158 ◽  
Author(s):  
MP Frant ◽  
M Lyjak ◽  
L Bocian ◽  
A Barszcz ◽  
K Niemczuk ◽  
...  
Keyword(s):  
Wild Boar ◽  
Active Surveillance ◽  
African Swine Fever Virus ◽  
Winter Season ◽  
African Swine Fever ◽  
Disease Spread ◽  
Passive Surveillance ◽  
Wild Boar Population ◽  
Wild Boars ◽  
In The Wild

African swine fever (ASF) was first described in 1921 in Kenya. The latest epidemic of ASF started in 2007 in Georgia. The virus was introduced to Poland in 2014. Since the beginning of the epidemics, the National Veterinary Research Institute in Pulawy (NVRI) has been testing wild boar samples from restricted areas and other parts of Poland to conduct passive and active surveillance for ASFV in these groups of animals. The aim of this study was to summarise the last two years of the ASF epidemiological status in Poland and the attempt to find disease patterns in the wild boar population. The period between 2017 and 2018 brought a massive number of new ASF cases in Poland. The number of ASF-positive wild boars jumped from 91 in 2016 to 1 140 in 2017 (approximately a 12 × increase), and 2018 was even worse, with the disease affecting 4 083 animals (2 435 cases; one case could even be 10 animals or more if they are found in one place next to each other). The percentage of positive wild boars found dead (passive surveillance) in the restricted area increased in 2018 to 73.1% from 70.8% in 2017. The chance of obtaining positive results in this group was six times higher in December and 4.5 times higher in January than in August and September. The percentage of positive wild boars detected through active surveillance reached 1.5% in 2018. The data suggested that, not only in Poland, but also in other ASF-affected countries, during the epizootic stage of the disease spread the most important measure is an effective passive surveillance of dead wild boars especially, in the winter season rather than in the summer.

scholarly journals What Do Lithuanian Hunters Think of African Swine Fever and Its Control—Perceptions

Animals ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 525
Author(s):  
Evelina Stončiūtė ◽  
Katja Schulz ◽  
Alvydas Malakauskas ◽  
Franz J. Conraths ◽  
Marius Masiulis ◽  
...  
Keyword(s):  
Wild Boar ◽  
Surveillance System ◽  
African Swine Fever ◽  
Study Data ◽  
Control Measures ◽  
Attitudes And Beliefs ◽  
Passive Surveillance ◽  
Wild Boar Population ◽  
In The Wild ◽  
Governmental Institutions

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.

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