Canine parvovirus (CPV) is a highly contagious and relatively common cause of acute, infectious GI illness in young dogs. Although its exact origin is unknown, it is believed to have arisen from feline panleukopenia virus or a related parvovirus of nondomestic animals
Parvo virus infection in dog - preventive medicinerajboy19
Canine parvovirus is a highly contagious infection of dogs caused by a Parvovirus. This slide include virology,transmission,pathogenesis,clinical sign and symptoms,diagnosis, differential diagnosis,treatment, prevention and control.
Feline vaccination is animal vaccination applied to cats. Vaccination plays a vital role in protecting cats from infectious diseases, some of which are potentially fatal. They can be exposed to these diseases from their environment, other pets, or even humans.
African Swine Fever: Nature, Impacts and Threats to the Global Pig Industry Garry D. Lasaga
In August 2018, African Swine Fever (ASF), one of the world’s most feared swine infection made headlines as it hit for the first time ever, the world’s largest pig producer – China. This review paper summarizes the current state of knowledge and very recent updates on ASF.
local names, definition, etiology,epidemiology lifecycle, pathogenesis, clinical findings, necropsy finding, diagnosis,treatment, control and prevention
etiology, local names, definition, transmission, source of infection, epidemiology, pathogenesis, clinical signs, diagnosis, differential diagnosis, treatment prevention and control
There are hundreds of diseases of livestock and pet animals that can be printed through properly used quality vaccines. This presentation summarises different types of vaccines used by veterinarians to control/ prevent diseases. The presentation enlists the vaccine-preventable diseases of pets and livestock, and also the different vaccines used.
Bovine herpesvirus 1 (BHV-1) is associated with several diseases in cattle: infectious bovine rhinotracheitis (IBR), infectious pustular vulvovaginitis (IPV), balanoposthitis, conjunctivitis, abortion, encephalomyelitis, and mastitis.
Canine parvovirus (CPV) is a highly contagious and relatively common cause of acute, infectious GI illness in young dogs. Although its exact origin is unknown, it is believed to have arisen from feline panleukopenia virus or a related parvovirus of nondomestic animals. It is a nonenveloped, single-stranded DNA virus, resistant to many common detergents and disinfectants, as well as to changes in temperature and pH. Infectious CPV can persist indoors at room temperature for at least 2 mo; outdoors, if protected from sunlight and desiccation, it can persist for many months and possibly years.
Canine parvovirus (CPV) is a highly contagious and relatively common cause of acute, infectious GI illness in young dogs. Although its exact origin is unknown, it is believed to have arisen from feline panleukopenia virus or a related parvovirus of nondomestic animals
Parvo virus infection in dog - preventive medicinerajboy19
Canine parvovirus is a highly contagious infection of dogs caused by a Parvovirus. This slide include virology,transmission,pathogenesis,clinical sign and symptoms,diagnosis, differential diagnosis,treatment, prevention and control.
Feline vaccination is animal vaccination applied to cats. Vaccination plays a vital role in protecting cats from infectious diseases, some of which are potentially fatal. They can be exposed to these diseases from their environment, other pets, or even humans.
African Swine Fever: Nature, Impacts and Threats to the Global Pig Industry Garry D. Lasaga
In August 2018, African Swine Fever (ASF), one of the world’s most feared swine infection made headlines as it hit for the first time ever, the world’s largest pig producer – China. This review paper summarizes the current state of knowledge and very recent updates on ASF.
local names, definition, etiology,epidemiology lifecycle, pathogenesis, clinical findings, necropsy finding, diagnosis,treatment, control and prevention
etiology, local names, definition, transmission, source of infection, epidemiology, pathogenesis, clinical signs, diagnosis, differential diagnosis, treatment prevention and control
There are hundreds of diseases of livestock and pet animals that can be printed through properly used quality vaccines. This presentation summarises different types of vaccines used by veterinarians to control/ prevent diseases. The presentation enlists the vaccine-preventable diseases of pets and livestock, and also the different vaccines used.
Bovine herpesvirus 1 (BHV-1) is associated with several diseases in cattle: infectious bovine rhinotracheitis (IBR), infectious pustular vulvovaginitis (IPV), balanoposthitis, conjunctivitis, abortion, encephalomyelitis, and mastitis.
Canine parvovirus (CPV) is a highly contagious and relatively common cause of acute, infectious GI illness in young dogs. Although its exact origin is unknown, it is believed to have arisen from feline panleukopenia virus or a related parvovirus of nondomestic animals. It is a nonenveloped, single-stranded DNA virus, resistant to many common detergents and disinfectants, as well as to changes in temperature and pH. Infectious CPV can persist indoors at room temperature for at least 2 mo; outdoors, if protected from sunlight and desiccation, it can persist for many months and possibly years.
African Swine Fever (ASF) virus genomics and diagnosticsILRI
Presented by Richard Bishop and Cynthia Onzere at the Closing workshop of the BecA‐ILRI‐CSIRO‐AusAID project on Understanding ASF epidemiology as a basis for control, Nairobi, Kenya, 2‐3 October 2013
A Novel Sheeppox virus vectored vaccine for Foot-and-Mouth-Disease was developed from scratch. The vaccine was tested in Sheep and found to elicit strong Antibody and Cell mediated Immune response when used in a prime boost regimen with Protein and DNA vaccines.
a) Describe two ways the researcher could minimise experimenter bias i.docxbickerstaffinell
a) Describe two ways the researcher could minimise experimenter bias in this study.
b) Describe a way the researchers could minimise sample bias in this study. (distinct from your answers in part a)
Nuclear import of SARS-CoV-2 nucleocapsid (N) protein is not inhibited by overmectin B.A. Loney* and M.A. Larkey* *Bundoora Institute for Applied Medical Research. Introduction The causative agent of the current COVID-19 pandemic, SARS-CoV-2, is a single stranded positive sense RNA virus that is closely related to severe acute respiratory syndrome coronavirus (SARS-CoV). It has been previously shown that SARS-CoV-2 nucleocapsid (N) is present in the cytoplasm but can also actively localize to the nucleolus where it can interact with host proteins and also bind to viral RNA. It has also been previously shown that nuclear import of similar nucleocapsid proteins (including the HIV-1 nucleocapsid protein, NC) from other RNA viruses can be inhibited by the drug overmectin resulting in decreased viral replication efficiency. There are multiple nuclear import pathways mediated by different receptors. The two most common nuclear import pathways are mediated by the importin / heterodimer or by the importin homodimer. Overmectin has previously been demonstrated to inhibit nuclear import by disrupting the importin / pathway. In this study SARS-CoV-2 nucleocapsid (N) was transfected into Hela cells and the effectiveness of overmectin at inhibiting its nuclear import was determined. Methods Expression of N protein in the absence of other viral proteins. To investigate the nuclear import of SARS-CoV-2 N protein three constructs were created. 1. pEGFP-NCov2. The N gene (from SARS-CoV-2, isolate BJ04) was cloned into the eukaryotic expression vector pEGFP-C1 (Promega) such that expression of the N gene was under the control of a cytomegalovirus (CMV) polymerase Il promoter to express an N protein fusion with the C-terminal of EGFP. 2. pEGFP. The pEGFP-C1 expression vector alone. 3. EEGFP-TRF1. A positive control for nuclear import. The human TRF1 gene was cloned into the eukaryotic expression vector pEGFP-C1 to express an N protein fusion with the C-terminal of EGFP. TRF1 nuclear localisation has shown to be mediated by the importin homodimer. Hela cells were cultured in 12 separate culture plates at a density of 1 0 5 cells per 9.6 cm 2 plate with each plate containing 2 coverslips. Cells were cultured using Cell Biologics' Culture Complete Growth Medium with 5\% foetal calf serum at 3 7 C and 5% CO 2 . Cells were transfected with 2 g of either pEGFPNCov2 (plates 1,4,7 and 10), pEGFP (plates 2, 5, 8 and 11) or pEGFP-TRF1 (3, 6, 9 and 12) and 50 g of Lipofectamine (GibcoBRL). 12 hours post-transfection, even numbered plates were treated with 5 M overmectin in DMSO and odd numbered plates were left untreated. After 24 hours coverslips were removed, and the cells fixed. DAPI was added to visualise the nuclei and the localisation of GFP determined by fluorescent mic.
Small ruminant keepers’ knowledge, attitudes and practices towards peste des ...ILRI
Presentation by Guy Ilboudo, Abel Sènabgè Biguezoton, Cheick Abou Kounta Sidibé, Modou Moustapha Lo, Zoë Campbell and Michel Dione at the 6th Peste des Petits Ruminants Global Research and Expertise Networks (PPR-GREN) annual meeting, Bengaluru, India, 28–30 November 2023.
Small ruminant keepers’ knowledge, attitudes and practices towards peste des ...ILRI
Poster by Guy Ilboudo, Abel Sènabgè Biguezoton, Cheick Abou Kounta Sidibé, Modou Moustapha Lo, Zoë Campbell and Michel Dione presented at the 6th Peste des Petits Ruminants Global Research and Expertise Networks (PPR-GREN) annual meeting, Bengaluru, India, 29 November 2023.
A training, certification and marketing scheme for informal dairy vendors in ...ILRI
Presentation by Silvia Alonso, Jef L. Leroy, Emmanuel Muunda, Moira Donahue Angel, Emily Kilonzi, Giordano Palloni, Gideon Kiarie, Paula Dominguez-Salas and Delia Grace at the Micronutrient Forum 6th Global Conference, The Hague, Netherlands, 16 October 2023.
Milk safety and child nutrition impacts of the MoreMilk training, certificati...ILRI
Poster by Silvia Alonso, Emmanuel Muunda, Moira Donahue Angel, Emily Kilonzi, Giordano Palloni, Gideon Kiarie, Paula Dominguez-Salas, Delia Grace and Jef L. Leroy presented at the Micronutrient Forum 6th Global Conference, The Hague, Netherlands, 16 October 2023.
Food safety research in low- and middle-income countriesILRI
Presentation by Hung Nguyen-Viet at the first technical meeting to launch the Food Safety Working Group under the One Health Partnership framework, Hanoi, Vietnam, 28 September 2023
Presentation by Hung Nguyen-Viet at the first technical meeting to launch the Food Safety Working Group under the One Health Partnership framework, Hanoi, Vietnam, 28 September 2023
Reservoirs of pathogenic Leptospira species in UgandaILRI
Presentation by Lordrick Alinaitwe, Martin Wainaina, Salome Dürr, Clovice Kankya, Velma Kivali, James Bugeza, Martin Richter, Kristina Roesel, Annie Cook and Anne Mayer-Scholl at the University of Bern Graduate School for Cellular and Biomedical Sciences Symposium, Bern, Switzerland, 29 June 2023.
Assessing meat microbiological safety and associated handling practices in bu...ILRI
Presentation by Patricia Koech, Winnie Ogutu, Linnet Ochieng, Delia Grace, George Gitao, Lily Bebora, Max Korir, Florence Mutua and Arshnee Moodley at the 8th All Africa Conference on Animal Agriculture, Gaborone, Botswana, 26–29 September 2023.
Ecological factors associated with abundance and distribution of mosquito vec...ILRI
Poster by Max Korir, Joel Lutomiah and Bernard Bett presented the 8th All Africa Conference on Animal Agriculture, Gaborone, Botswana, 26–29 September 2023.
Practices and drivers of antibiotic use in Kenyan smallholder dairy farmsILRI
Poster by Lydiah Kisoo, Dishon M. Muloi, Walter Oguta, Daisy Ronoh, Lynn Kirwa, James Akoko, Eric Fèvre, Arshnee Moodley and Lillian Wambua presented at Tropentag 2023, Berlin, Germany, 20–22 September 2023.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Professional air quality monitoring systems provide immediate, on-site data for analysis, compliance, and decision-making.
Monitor common gases, weather parameters, particulates.
Seminar of U.V. Spectroscopy by SAMIR PANDASAMIR PANDA
Spectroscopy is a branch of science dealing the study of interaction of electromagnetic radiation with matter.
Ultraviolet-visible spectroscopy refers to absorption spectroscopy or reflect spectroscopy in the UV-VIS spectral region.
Ultraviolet-visible spectroscopy is an analytical method that can measure the amount of light received by the analyte.
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
DERIVATION OF MODIFIED BERNOULLI EQUATION WITH VISCOUS EFFECTS AND TERMINAL V...Wasswaderrick3
In this book, we use conservation of energy techniques on a fluid element to derive the Modified Bernoulli equation of flow with viscous or friction effects. We derive the general equation of flow/ velocity and then from this we derive the Pouiselle flow equation, the transition flow equation and the turbulent flow equation. In the situations where there are no viscous effects , the equation reduces to the Bernoulli equation. From experimental results, we are able to include other terms in the Bernoulli equation. We also look at cases where pressure gradients exist. We use the Modified Bernoulli equation to derive equations of flow rate for pipes of different cross sectional areas connected together. We also extend our techniques of energy conservation to a sphere falling in a viscous medium under the effect of gravity. We demonstrate Stokes equation of terminal velocity and turbulent flow equation. We look at a way of calculating the time taken for a body to fall in a viscous medium. We also look at the general equation of terminal velocity.
Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
on Io’s surface have been monitored from both spacecraft and ground-based telescopes.
Here, we present the highest spatial resolution images of Io ever obtained from a groundbased telescope. These images, acquired by the SHARK-VIS instrument on the Large
Binocular Telescope, show evidence of a major resurfacing event on Io’s trailing hemisphere. When compared to the most recent spacecraft images, the SHARK-VIS images
show that a plume deposit from a powerful eruption at Pillan Patera has covered part
of the long-lived Pele plume deposit. Although this type of resurfacing event may be common on Io, few have been detected due to the rarity of spacecraft visits and the previously low spatial resolution available from Earth-based telescopes. The SHARK-VIS instrument ushers in a new era of high resolution imaging of Io’s surface using adaptive
optics at visible wavelengths.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
(May 29th, 2024) Advancements in Intravital Microscopy- Insights for Preclini...Scintica Instrumentation
Intravital microscopy (IVM) is a powerful tool utilized to study cellular behavior over time and space in vivo. Much of our understanding of cell biology has been accomplished using various in vitro and ex vivo methods; however, these studies do not necessarily reflect the natural dynamics of biological processes. Unlike traditional cell culture or fixed tissue imaging, IVM allows for the ultra-fast high-resolution imaging of cellular processes over time and space and were studied in its natural environment. Real-time visualization of biological processes in the context of an intact organism helps maintain physiological relevance and provide insights into the progression of disease, response to treatments or developmental processes.
In this webinar we give an overview of advanced applications of the IVM system in preclinical research. IVIM technology is a provider of all-in-one intravital microscopy systems and solutions optimized for in vivo imaging of live animal models at sub-micron resolution. The system’s unique features and user-friendly software enables researchers to probe fast dynamic biological processes such as immune cell tracking, cell-cell interaction as well as vascularization and tumor metastasis with exceptional detail. This webinar will also give an overview of IVM being utilized in drug development, offering a view into the intricate interaction between drugs/nanoparticles and tissues in vivo and allows for the evaluation of therapeutic intervention in a variety of tissues and organs. This interdisciplinary collaboration continues to drive the advancements of novel therapeutic strategies.
Salas, V. (2024) "John of St. Thomas (Poinsot) on the Science of Sacred Theol...Studia Poinsotiana
I Introduction
II Subalternation and Theology
III Theology and Dogmatic Declarations
IV The Mixed Principles of Theology
V Virtual Revelation: The Unity of Theology
VI Theology as a Natural Science
VII Theology’s Certitude
VIII Conclusion
Notes
Bibliography
All the contents are fully attributable to the author, Doctor Victor Salas. Should you wish to get this text republished, get in touch with the author or the editorial committee of the Studia Poinsotiana. Insofar as possible, we will be happy to broker your contact.
Toxic effects of heavy metals : Lead and Arsenicsanjana502982
Heavy metals are naturally occuring metallic chemical elements that have relatively high density, and are toxic at even low concentrations. All toxic metals are termed as heavy metals irrespective of their atomic mass and density, eg. arsenic, lead, mercury, cadmium, thallium, chromium, etc.
1. Lucilla Steinaa, Principal Scientist, ILRI
International Symposium on Animal Functional Genomics & Health
19 Nov 2020
Towards a vaccine for African swine fever
2. • Caused by a large double stranded DNA
virus.
• Between 150-167 genes, 22 genotypes
based on the p72 sequence.
• Belongs to the family Asfaviridae, nearest
relatives are the poxviruses.
• There is a wildlife reservoir : warthogs,
bush pigs. Soft ticks are involved in
transmission.
African swine fever
3. • ASF is now a huge problem in Eastern
Europe, Russia, China, Asia, and Africa.
• ASF is present in about 26 countries in
Africa. Multiple genotypes are present.
• ASF is a major problem for potential pig
farmers, who are discouraged from
entering the pig business due to ASF.
African swine fever
5. Neighbor-Joining tree depicting the p72 gene relationships
and geographical distribution of the major ASFV genotypes
CAM/1/02
CAM/4/85
GHA/1/02
LIS/60
NIG/1/01
ANG/70
IC/2/96
BEN/1/97
SPEC/205
SPEC/209
Mkuzi/79
I
MOZ/1/02
MOZ/1/05
LUS1/93
MOZ/2/02
MOZ/1/03
II
RSA/5/95
SPEC/257
RSA/3/03
RSA/Warmbaths
III
MOZ/1979
MAL/1/02
MOZ/1960
Tengani/62
V
MOZ/8/94
SPEC265
MOZ/1/94
VI
RSA/3/04/
RSA/Warthog
RSA/1/99/W
IV
RSA/2/96
RSA/2/03
RSA/3/96
RSA/1/04
XIX
Lillie
RSA/1/95
XX
XIIIRSA/1/03
RSA/4/95
RSA/1/98
SPEC/154
VII
XIVNYA/12
TAN/2/03
TAN/1/03
XVI
XVTAN/1/01
XIIISUM/14/11
XIIMZI/1/92
XIKAB/6/2
MCH/1/89
MCH/3/89
BAN/1/91
DED/1/89
LIL/20/2
DOWA
CHJ/1/89
KLI/2/88
PHW/1/88
TMB/1/89
VIII
UGA/1/95
KEN/5/01 IX
BUR/1/84
BUR/1/90
Kenya/50
X
100
79
74
57
91
0.005
p72 gene
Contribution: Livio Heath (ARC-OVI)
Distribution of genotypes in Africa
Genotype I
Genotype II,III, IV, V,
VI, VII, XIX, XX, XIII
Genotype VIII, XI, XII, XIII,
XV, XVI, XIV
Genotype IX, X
6. Engineering of African Swine Fever Virus
for Production of Live Attenuated Vaccine
Candidates
7. Engineering of African Swine Fever Virus for
Production of Live Attenuated Vaccine Candidates
• Establishment of vaccine platforms for making attenuated
African swine fever vaccine candidates
• Gene modification by CRISPR-Cas.
• Gene modification by using a synthetic approach – reverse genetics system
• International Livestock Research Institute, J. Craig Venter Institute and
Friedrich Loeffler Institute.
8. • The virus
• Genotype IX isolated from an outbreak area
between Uganda and Kenya. Genomic
sequence available.
• Viruses from current outbreak areas in the
region (Kenya, Uganda, (Tanzania)) have been
sequenced at ILRI. All are genotype IX and X,
and genomes are almost identical to the one
we have. Genotype IX and X have been
detected in outbreaks in Congo.
Virus backbone for modification of ASFV
Gallardo C et al. A.J. Biotech 2011
Onzere C. et al. Virus Genes 2018
10. CRISPR-modification pathway (NHEJ)
Stable CAS
transfected cell line
(WSL)
Transfection with
guide RNA
Infect with
GFPvirus
Harvestsup./cells
with virus
10-1
10-2 10-3
10-4
10-5 10-6
Checkfor genome
modifications
Dilutions
Virusclones
Genome cleavage
detection kit
OR
DNA Sequencing of
modified region
11. Genome cleavage detection
uncleaved
cleaved
Mismatched
heterogenous
duplex
Denature &
Reanneal
GeneArt™GenomicCleavageDetection Kit
Digestw/ detection
enzyme
Cleaved
heteroduplex
Electrophoresis
T C
T-Treated
C- Control
PCRamplify
targeted locus
Banddensitometryfor
cleavageefficiency
G1 G2 C
1 2 3
500
PCRamplify
targetedlocus
1 2 3
Denature& Reanneal
Digest w/ detectionenzyme
Electrophoresis
CRISP-modification of two genes (SingleguideRNA)
500
300
200
1 2 3 1 2 3
negative enzyme
control
13. A clone virus with desired mutation
1 2 3 1 2 3
gRNA target
Stop Codon
14. 17th Oct 2019
Reflections on the NHEJ pathway
• Bias towards in-frame indels using NHEJ, one guide RNA approach.
• Difficulties in cloning due to lack of marker for modification.
Mitigation:
• Use HDR approach
NHEJ pathway
16. GFP plasmid pGFPΔA238L for integration of GFP
in the ASF genome
17th Oct 2019
pGFPΔA238L, synthesised by GenScript
circular
linear
Infection /transfection experiments in WSL– Cas cell line
Kenya 1033 (WT) + plasmid
Kenya 1033 (WT) + amplicon
Kenya 1033 (∆CD2v) + plasmid
Kenya 1033 (∆CD2v) + amplicon
18. PCR confirmation of GFP genome integration
17th Oct 2019
Amplicon sizes:
F1R1 (5′) = 1488 bp
F1R2 = 3740 bp (KO),
F1R2 = 3337 bp (wt),
F2R2 (3′) = 1681 bp
Expected band sizes for both amplicon and plasmid
transfections in Kenya1033 & Kenya1033-ΔCD2v
Kenya1033
amplicon
Kenya1033
plasmid
1033-ΔCD2v
plasmid
1033-ΔCD2v
amplicon
F1R1
F2R2
F1R2
F1R1
F2R2
F1R2
F1R1
F2R2
F1R2
F1R1
F2R2
F1R2
5Kb
3Kb
1.5Kb
PCR to check integration of the plasmid/amplicon into the virus genome
F1 R1 F2 R2
21. Toward a subunit vaccine
• Identification of antigens for use in a subunit vaccine
• Screening of peptide library in ELISpot
• Production of recombinant viral vectors
• Adenovirus/MVA prime-boost
• Testing of pools of viral vectors in pigs for protection
Adenovirus Vaccinia
24. CD2v knockout
• The virus is attenuated.
• 80-90% of pigs survives 103 HAD.
• Clearly, another mutation is needed.
• We are soon testing if pigs immunized
with ASF1033ΔCD2v are protected
against wild-type.
26. Antibody responses in pigs after
immunization with attenuated ASFΔCD2v
0
20
40
60
80
100
120
0 5 10 15 20 25 30
PB188 PB190 PB191 PB192 PB195 PB196 PB197
Days post infection
%inhibition
Cut off
27. Antibody responses in pigs after
immunization with attenuated ASFΔCD2v
D0D28 D0D28 D0D28
0
50
100
%Blocking
Local breed
European breed II
European breed I
28. Recognition by PBMC of overlapping peptides
spanning the entire proteome
immunization with attenuated ASFΔCD2v
• 217 gene pools in all
• 42 pools recognized in all 3
screenings.
• 62 pools not recognized
Genepool
29. Peptide pools recognized in ELISpot using full
proteome library
immunization with attenuated ASFΔCD2v
CD8 cells
PBMC
30. Single antigen recognition by number of animals
CD8 cells
Best recognition
of single Ag:
6/22 animals
PBMC
Best recognition
of single Ag:
17/22 animals
31. Production of viral vectors with ASF genes
• Ten genes from the peptide
screen were selected
• Each of the 10 genes
produced in both
adenovirus (HuAd5) and
Modified Vaccinia virus
Ankara (MVA).
• Next, to test pools in the
animal model
32. Virus batch for challenge experiments
0 2 4 6 8 10
0
50
100
Survival - Groups
Day after challenge
Percentsurvival
102
103
104
105
0 2 4 6 8
38
40
42
Body temperature - Groups
Day after challenge
Bodytemperature
102
103
104
105
0 2 4 6 8 10
0
5
10
15
Clinical score - Groups
Day after challenge
Clinicalscore
102
103
104
105
Establishment of minimal lethal challenging dose
HAD-titer: 102, 103, 104, 105
5 animals per group
Scoring system: King et al.
33. 33
ILRI
Hussein Abkallo
Hanneke Hemmink
Nicholas Svitek
Jeremiah Khayumba
Anna Lacasta
Elias Awino
Rosemary Saya
Bernard Odour
Emanuel Khazalwa
Lucilla Steinaa
Collaborators
Sanjay Vashee, Assoc prof.
J.Craig Venter Instisitute
Walter Fuchs, Group leader
Friedrich Loeffler Institute
Acknowledgements
34. This presentation is licensed for use under the Creative Commons Attribution 4.0 International Licence.
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