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Research paper thumbnail of Participatory assessment of risk factors for African swine fever in western Kenya

Research paper thumbnail of High Doses of Inactivated African Swine Fever Virus Are Safe, but Do Not Confer Protection against a Virulent Challenge

Vaccines, 2021

African swine fever (ASF) is currently the major concern of the global swine industry, as a conse... more African swine fever (ASF) is currently the major concern of the global swine industry, as a consequence of which a reconsideration of the containment and prevention measures taken to date is urgently required. A great interest in developing an effective and safe vaccine against ASF virus (ASFV) infection has, therefore, recently appeared. The objective of the present study is to test an inactivated ASFV preparation under a vaccination strategy that has not previously been tested in order to improve its protective effect. The following have been considered: (i) virus inactivation by using a low binary ethyleneimine (BEI) concentration at a low temperature, (ii) the use of new and strong adjuvants; (iii) the use of very high doses (6 × 109 haemadsorption in 50% of infected cultures (HAD50)), and (iv) simultaneous double inoculation by two different routes of administration: intradermal and intramuscular. Five groups of pigs were, therefore, inoculated with BEI- Pol16/DP/OUT21 in diffe...

Research paper thumbnail of African swine fever (ASF) diagnosis, an essential tool in the epidemiological investigation

Research paper thumbnail of First Oral Vaccination of Eurasian Wild Boar Against African Swine Fever Virus Genotype II

Frontiers in Veterinary Science, 2019

Research paper thumbnail of Detection of African Swine Fever Virus Antibodies in Serum and Oral Fluid Specimens Using a Recombinant Protein 30 (p30) Dual Matrix Indirect ELISA

Research paper thumbnail of Phylogenomic analysis of 11 complete African swine fever virus genome sequences

Research paper thumbnail of Enhanced discrimination of African swine fever virus isolates through nucleotide sequencing of the p54, p72, and pB602L (CVR) genes

Research paper thumbnail of Genetic characterisation of African swine fever viruses from recent and historical outbreaks in Sardinia (1978–2009)

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

Transboundary and Emerging Diseases, 2012

Research paper thumbnail of Risk factors for farm-level African swine fever infection in major pig-producing areas in Nigeria, 1997–2011

Preventive Veterinary Medicine, 2012

Research paper thumbnail of Development and inter-laboratory validation study of an improved new real-time PCR assay with internal control for detection and laboratory diagnosis of African swine fever virus

Journal of Virological Methods, 2011

Research paper thumbnail of Gaps in African swine fever: Analysis and priorities

Transboundary and Emerging Diseases, 2017

Research paper thumbnail of Disruption of nuclear organization during the initial phase of African swine fever virus infection

African swine fever virus (ASFV), the causative agent of one of the most devastating swine diseas... more African swine fever virus (ASFV), the causative agent of one of the most devastating swine diseases, has been considered exclusively cytoplasmic, even though some authors have shown evidence of an early stage of nuclear replication. In the present study, an increment of lamin A/C phosphorylation was observed in ASFV-infected cells as early as 4 h postinfection, followed by the disassembling of the lamina network close to the sites where the viral genome starts its replication. At later time points, this and other nuclear envelope markers were found in the cytoplasm of the infected cells. The effect of the infection on the cell nucleus was much more severe than previously expected, since a redistribution of other nuclear proteins, such as RNA polymerase II, the splicing speckle SC-35 marker, and the B-23 nucleolar marker, was observed from 4 h postinfection. All this evidence, together with the redistribution, dephosphorylation, and subsequent degradation of RNA polymerase II after A...

Research paper thumbnail of Effect of mixing and feed batch sequencing on the prevalence and distribution of African swine fever virus in swine feed

Transboundary and Emerging Diseases

It is critical to have methods that can detect and mitigate the risk of African swine fever virus... more It is critical to have methods that can detect and mitigate the risk of African swine fever virus (ASFV) in potentially contaminated feed or ingredients bound for the United States. This work's purpose was to evaluate feed batch sequencing as a mitigation technique for ASFV contamination in a feed mill and to determine if a feed sampling method could identify ASFV following experimental inoculation. Batches of feed were manufactured in a BSL-3Ag room at Kansas State University's Biosafety Research Institute in Manhattan, Kansas. First, the pilot feed manufacturing system mixed, conveyed, and discharged an ASFV-free diet. Next, a diet was manufactured using the same equipment, but contained feed inoculated with ASFV for final concentration of 5.6 × 104 TCID50 /g. Then, four subsequent ASFV-free batches of feed were manufactured. After discharging each batch into a collection container, 10 samples were collected in a double 'X' pattern. Samples were analyzed using a qPCR assay for ASFV p72 gene then the cycle threshold (Ct) and Log10 genomic copy number (CN)/g of feed were determined. Batch of feed impacted the qPCR Ct values (P < 0.0001) and the Log10 genomic CN/g (P < 0.0001) content of feed samples. Feed samples obtained after manufacturing the ASFV contaminated diet contained the greatest amounts of ASFV p72 DNA across all criteria (P < 0.05). Quantity of ASFV p72 DNA decreased sequentially as additional batches of feed were manufactured but was still detectable after batch sequence 4. This subsampling method was able to identify ASFV genetic material in feed samples using p72 qPCR. In summary, sequencing batches of feed decreases concentration of ASFV contamination in feed, but does not eliminate it. Bulk ingredients can be accurately evaluated for ASFV contamination by collecting 10 subsamples using the sampling method described herein. Future research is needed to evaluate if different mitigation techniques can reduce ASFV feed contamination. This article is protected by copyright. All rights reserved.

Research paper thumbnail of Prevalence and Distribution of African Swine Fever Virus in Swine Feed After Mixing and Feed Batch Sequencing

Kansas Agricultural Experiment Station Research Reports

Research paper thumbnail of Identification of a New Genotype of African Swine Fever Virus in Domestic Pigs from Ethiopia

Transboundary and Emerging Diseases, 2016

African swine fever (ASF) is an important emerging transboundary animal disease (TAD), which curr... more African swine fever (ASF) is an important emerging transboundary animal disease (TAD), which currently has an impact on many countries in Africa, Eastern Europe, the Caucasus and the Russian Federation. The current situation in Europe shows the ability of the virus to rapidly spread, which stands to threaten the global swine industry. At present, there is no viable vaccine to minimize spread of the disease and stamping out is the main source of control. In February 2011, Ethiopia had reported its first suspected outbreaks of ASF. Genomic analyses of the collected ASF virus (ASFV) strains were undertaken using 23 tissue samples collected from domestic swine in Ethiopia from 2011 to 2014. The analysis of Ethiopian ASFVs partial p72 gene sequence showed the identification of a new genotype, genotype XXIII, that shares a common ancestor with genotypes IX and X, which comprise isolates circulating in Eastern African countries and the Republic of Congo. Analysis of the p54 gene also followed the p72 pattern and the deduced amino acid sequence of the central variable region (CVR) of the B602L gene showed novel tetramer repeats not previously characterized.

Research paper thumbnail of African Swine Fever Virus p72 Genotype IX in Domestic Pigs, Congo, 2009

Emerging infectious …, 2011

Research paper thumbnail of Surveillance for African Swine Fever in Nigeria, 2006–2009

Transboundary and …, 2010

African swine fever (ASF) has had significant economic and social impact in Nigeria since 1997. H... more African swine fever (ASF) has had significant economic and social impact in Nigeria since 1997. However, there has been no effective national response to bring it under control. In this report, we confirm that ASF is still prevalent and widespread in Nigeria. Results from both ...

Research paper thumbnail of Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How to Improve Surveillance and Control Programmes

Journal of Clinical Microbiology, 2015

This study represents a complete comparative analysis of the most widely used African swine fever... more This study represents a complete comparative analysis of the most widely used African swine fever (ASF) diagnostic techniques in the European Union (EU) using field and experimental samples from animals infected with genotype II ASF virus (ASFV) isolates circulating in Europe. To detect ASFV, three different PCRs were evaluated in parallel using 785 field and experimental samples. The results showed almost perfect agreement between the Universal ProbeLibrary (UPL-PCR) and the real-time (κ = 0.94 [95% confidence interval {CI}, 0.91 to 0.97]) and conventional (κ = 0.88 [95% CI, 0.83 to 0.92]) World Organisation for Animal Health (OIE)-prescribed PCRs. The UPL-PCR had greater diagnostic sensitivity for detecting survivors and allows earlier detection of the disease. Compared to the commercial antigen enzyme-linked immunosorbent assay (ELISA), good-to-moderate agreement (κ = 0.67 [95% CI, 0.58 to 0.76]) was obtained, with a sensitivity of 77.2% in the commercial test. For ASF antibody d...

Research paper thumbnail of Dynamics of African swine fever virus (ASFV) infection in domestic pigs infected with virulent, moderate virulent and attenuated genotype II ASFV European isolates

Transboundary and Emerging Diseases

Research paper thumbnail of Participatory assessment of risk factors for African swine fever in western Kenya

Research paper thumbnail of High Doses of Inactivated African Swine Fever Virus Are Safe, but Do Not Confer Protection against a Virulent Challenge

Vaccines, 2021

African swine fever (ASF) is currently the major concern of the global swine industry, as a conse... more African swine fever (ASF) is currently the major concern of the global swine industry, as a consequence of which a reconsideration of the containment and prevention measures taken to date is urgently required. A great interest in developing an effective and safe vaccine against ASF virus (ASFV) infection has, therefore, recently appeared. The objective of the present study is to test an inactivated ASFV preparation under a vaccination strategy that has not previously been tested in order to improve its protective effect. The following have been considered: (i) virus inactivation by using a low binary ethyleneimine (BEI) concentration at a low temperature, (ii) the use of new and strong adjuvants; (iii) the use of very high doses (6 × 109 haemadsorption in 50% of infected cultures (HAD50)), and (iv) simultaneous double inoculation by two different routes of administration: intradermal and intramuscular. Five groups of pigs were, therefore, inoculated with BEI- Pol16/DP/OUT21 in diffe...

Research paper thumbnail of African swine fever (ASF) diagnosis, an essential tool in the epidemiological investigation

Research paper thumbnail of First Oral Vaccination of Eurasian Wild Boar Against African Swine Fever Virus Genotype II

Frontiers in Veterinary Science, 2019

Research paper thumbnail of Detection of African Swine Fever Virus Antibodies in Serum and Oral Fluid Specimens Using a Recombinant Protein 30 (p30) Dual Matrix Indirect ELISA

Research paper thumbnail of Phylogenomic analysis of 11 complete African swine fever virus genome sequences

Research paper thumbnail of Enhanced discrimination of African swine fever virus isolates through nucleotide sequencing of the p54, p72, and pB602L (CVR) genes

Research paper thumbnail of Genetic characterisation of African swine fever viruses from recent and historical outbreaks in Sardinia (1978–2009)

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

Transboundary and Emerging Diseases, 2012

Research paper thumbnail of Risk factors for farm-level African swine fever infection in major pig-producing areas in Nigeria, 1997–2011

Preventive Veterinary Medicine, 2012

Research paper thumbnail of Development and inter-laboratory validation study of an improved new real-time PCR assay with internal control for detection and laboratory diagnosis of African swine fever virus

Journal of Virological Methods, 2011

Research paper thumbnail of Gaps in African swine fever: Analysis and priorities

Transboundary and Emerging Diseases, 2017

Research paper thumbnail of Disruption of nuclear organization during the initial phase of African swine fever virus infection

African swine fever virus (ASFV), the causative agent of one of the most devastating swine diseas... more African swine fever virus (ASFV), the causative agent of one of the most devastating swine diseases, has been considered exclusively cytoplasmic, even though some authors have shown evidence of an early stage of nuclear replication. In the present study, an increment of lamin A/C phosphorylation was observed in ASFV-infected cells as early as 4 h postinfection, followed by the disassembling of the lamina network close to the sites where the viral genome starts its replication. At later time points, this and other nuclear envelope markers were found in the cytoplasm of the infected cells. The effect of the infection on the cell nucleus was much more severe than previously expected, since a redistribution of other nuclear proteins, such as RNA polymerase II, the splicing speckle SC-35 marker, and the B-23 nucleolar marker, was observed from 4 h postinfection. All this evidence, together with the redistribution, dephosphorylation, and subsequent degradation of RNA polymerase II after A...

Research paper thumbnail of Effect of mixing and feed batch sequencing on the prevalence and distribution of African swine fever virus in swine feed

Transboundary and Emerging Diseases

It is critical to have methods that can detect and mitigate the risk of African swine fever virus... more It is critical to have methods that can detect and mitigate the risk of African swine fever virus (ASFV) in potentially contaminated feed or ingredients bound for the United States. This work's purpose was to evaluate feed batch sequencing as a mitigation technique for ASFV contamination in a feed mill and to determine if a feed sampling method could identify ASFV following experimental inoculation. Batches of feed were manufactured in a BSL-3Ag room at Kansas State University's Biosafety Research Institute in Manhattan, Kansas. First, the pilot feed manufacturing system mixed, conveyed, and discharged an ASFV-free diet. Next, a diet was manufactured using the same equipment, but contained feed inoculated with ASFV for final concentration of 5.6 × 104 TCID50 /g. Then, four subsequent ASFV-free batches of feed were manufactured. After discharging each batch into a collection container, 10 samples were collected in a double 'X' pattern. Samples were analyzed using a qPCR assay for ASFV p72 gene then the cycle threshold (Ct) and Log10 genomic copy number (CN)/g of feed were determined. Batch of feed impacted the qPCR Ct values (P < 0.0001) and the Log10 genomic CN/g (P < 0.0001) content of feed samples. Feed samples obtained after manufacturing the ASFV contaminated diet contained the greatest amounts of ASFV p72 DNA across all criteria (P < 0.05). Quantity of ASFV p72 DNA decreased sequentially as additional batches of feed were manufactured but was still detectable after batch sequence 4. This subsampling method was able to identify ASFV genetic material in feed samples using p72 qPCR. In summary, sequencing batches of feed decreases concentration of ASFV contamination in feed, but does not eliminate it. Bulk ingredients can be accurately evaluated for ASFV contamination by collecting 10 subsamples using the sampling method described herein. Future research is needed to evaluate if different mitigation techniques can reduce ASFV feed contamination. This article is protected by copyright. All rights reserved.

Research paper thumbnail of Prevalence and Distribution of African Swine Fever Virus in Swine Feed After Mixing and Feed Batch Sequencing

Kansas Agricultural Experiment Station Research Reports

Research paper thumbnail of Identification of a New Genotype of African Swine Fever Virus in Domestic Pigs from Ethiopia

Transboundary and Emerging Diseases, 2016

African swine fever (ASF) is an important emerging transboundary animal disease (TAD), which curr... more African swine fever (ASF) is an important emerging transboundary animal disease (TAD), which currently has an impact on many countries in Africa, Eastern Europe, the Caucasus and the Russian Federation. The current situation in Europe shows the ability of the virus to rapidly spread, which stands to threaten the global swine industry. At present, there is no viable vaccine to minimize spread of the disease and stamping out is the main source of control. In February 2011, Ethiopia had reported its first suspected outbreaks of ASF. Genomic analyses of the collected ASF virus (ASFV) strains were undertaken using 23 tissue samples collected from domestic swine in Ethiopia from 2011 to 2014. The analysis of Ethiopian ASFVs partial p72 gene sequence showed the identification of a new genotype, genotype XXIII, that shares a common ancestor with genotypes IX and X, which comprise isolates circulating in Eastern African countries and the Republic of Congo. Analysis of the p54 gene also followed the p72 pattern and the deduced amino acid sequence of the central variable region (CVR) of the B602L gene showed novel tetramer repeats not previously characterized.

Research paper thumbnail of African Swine Fever Virus p72 Genotype IX in Domestic Pigs, Congo, 2009

Emerging infectious …, 2011

Research paper thumbnail of Surveillance for African Swine Fever in Nigeria, 2006–2009

Transboundary and …, 2010

African swine fever (ASF) has had significant economic and social impact in Nigeria since 1997. H... more African swine fever (ASF) has had significant economic and social impact in Nigeria since 1997. However, there has been no effective national response to bring it under control. In this report, we confirm that ASF is still prevalent and widespread in Nigeria. Results from both ...

Research paper thumbnail of Assessment of African Swine Fever Diagnostic Techniques as a Response to the Epidemic Outbreaks in Eastern European Union Countries: How to Improve Surveillance and Control Programmes

Journal of Clinical Microbiology, 2015

This study represents a complete comparative analysis of the most widely used African swine fever... more This study represents a complete comparative analysis of the most widely used African swine fever (ASF) diagnostic techniques in the European Union (EU) using field and experimental samples from animals infected with genotype II ASF virus (ASFV) isolates circulating in Europe. To detect ASFV, three different PCRs were evaluated in parallel using 785 field and experimental samples. The results showed almost perfect agreement between the Universal ProbeLibrary (UPL-PCR) and the real-time (κ = 0.94 [95% confidence interval {CI}, 0.91 to 0.97]) and conventional (κ = 0.88 [95% CI, 0.83 to 0.92]) World Organisation for Animal Health (OIE)-prescribed PCRs. The UPL-PCR had greater diagnostic sensitivity for detecting survivors and allows earlier detection of the disease. Compared to the commercial antigen enzyme-linked immunosorbent assay (ELISA), good-to-moderate agreement (κ = 0.67 [95% CI, 0.58 to 0.76]) was obtained, with a sensitivity of 77.2% in the commercial test. For ASF antibody d...

Research paper thumbnail of Dynamics of African swine fever virus (ASFV) infection in domestic pigs infected with virulent, moderate virulent and attenuated genotype II ASFV European isolates

Transboundary and Emerging Diseases