Journal: The Journal of Veterinary Medical Science

Article Title: Pathological lesions and presence of viral antigens in four surviving pigs in African swine fever outbreak farms in Vietnam

doi: 10.1292/jvms.21-0409

Figure Lengend Snippet: Hematoxylin & eosin and immunohistochemical staining for African swine fever phosphoprotein p30 in tonsils. The inset show African swine fever virus-positive cell. No remarkable changes were noticed in the tonsils of case no. 4 (a) . Positive staining of mononuclear cells was detected in apposition to the tonsillar crypt in case no. 4 (b) .

Article Snippet: The rabbit polyclonal ASFV phosphoprotein p30 antibody (Alpha Diagnostic International, San Antonio, TX, USA) was used as the primary antibody.

Techniques: Immunohistochemical staining, Staining, Virus

Journal: The Journal of Veterinary Medical Science

Article Title: Pathological lesions and presence of viral antigens in four surviving pigs in African swine fever outbreak farms in Vietnam

doi: 10.1292/jvms.21-0409

Figure Lengend Snippet: Hematoxylin & eosin and immunohistochemical staining for African swine fever phosphoprotein p30 in lymph nodes. Marked fibrosis predominantly presented in the sinuses, especially in the subcapsular area of case no. 2 (a) . Antigen-labeled cell was not present in the lesion of case no. 2 (b) .

Article Snippet: The rabbit polyclonal ASFV phosphoprotein p30 antibody (Alpha Diagnostic International, San Antonio, TX, USA) was used as the primary antibody.

Techniques: Immunohistochemical staining, Staining, Labeling

Journal: The Journal of Veterinary Medical Science

Article Title: Pathological lesions and presence of viral antigens in four surviving pigs in African swine fever outbreak farms in Vietnam

doi: 10.1292/jvms.21-0409

Figure Lengend Snippet: Hematoxylin & eosin and immunohistochemical staining for African swine fever phosphoprotein p30 in lungs. The inset show African swine fever virus-positive cell. Bronchus-associated lymphoid tissue hyperplasia appeared in case no. 2 (a) with positive cells (b) . Thickened alveolar septum due to fibrous tissue was apparent in case no. 3 (c) with presentation of viral antigen-labeled cells in the alveoli (d) .

Article Snippet: The rabbit polyclonal ASFV phosphoprotein p30 antibody (Alpha Diagnostic International, San Antonio, TX, USA) was used as the primary antibody.

Techniques: Immunohistochemical staining, Staining, Virus, Labeling

Journal: The Journal of Veterinary Medical Science

Article Title: Pathological lesions and presence of viral antigens in four surviving pigs in African swine fever outbreak farms in Vietnam

doi: 10.1292/jvms.21-0409

Figure Lengend Snippet: Immunohistochemical (IHC) staining for African swine fever (ASF) phosphoprotein p30 (a) and double IHC staining for ASF phosphoprotein p30 as the brown color with Iba-1 (b) , vimentin (c) , and AE1/AE3 (d) as the purplish grey color in stomach of case no. 4. The inset show African swine fever virus-positive cell. A positive reaction occurred in amorphous cells in the gastric glands (a). No double stained cells were observed (b–d).

Article Snippet: The rabbit polyclonal ASFV phosphoprotein p30 antibody (Alpha Diagnostic International, San Antonio, TX, USA) was used as the primary antibody.

Techniques: Immunohistochemical staining, Immunohistochemistry, Virus, Staining

Journal: Pathogens

Article Title: A Practical Framework for ASFV Disinfectant Evaluation: Differentiating Cytopathic Effects from Cytotoxicity via Integrated Analytical Methods

doi: 10.3390/pathogens14050451

Figure Lengend Snippet: Damage scores in Vero cells exposed to ASFV and disinfectant treatments. Bright-field images of Vero cells were captured following treatment with ASFV alone (A), disinfectant only (D), or ASFV plus disinfectant (AD) at various dilutions (175×, 275×, 375×) and observed daily from 1 to 6 days post inoculation (DPI). Images were acquired at a total magnification of 200× using an Olympus IX73 inverted microscope equipped with a 20×/0.40 objective. All images are shown at the same scale for comparative purposes; therefore, individual scale bars are not included. Scoring system for cell damage: 0: No visible damage; cells appear healthy; 1: Minimal damage; slight rounding or detachment of a few cells; 2: Mild damage; increased rounding and detachment of cells (~10–20%); 3: Moderate damage; significant rounding, detachment, and cell loss (~30–50%); 4: Severe damage; widespread cell rounding and detachment (~50–80%); 5: Complete damage; almost all cells are detached or destroyed (>80%).

Article Snippet: Cells were blocked with 5% fetal bovine serum (FBS) in PBS for 30 min and incubated overnight at 4 °C with a primary antibody specific to ASFV p54 (1:200 dilution in blocking buffer, GeneTex, Irvine, CA, USA).

Techniques: Inverted Microscopy

Journal: Pathogens

Article Title: A Practical Framework for ASFV Disinfectant Evaluation: Differentiating Cytopathic Effects from Cytotoxicity via Integrated Analytical Methods

doi: 10.3390/pathogens14050451

Figure Lengend Snippet: Antigen detection results for ASFV with different disinfectant treatments. ( A ) Lateral flow assay (LFA) results for ASFV detection after treatment with various disinfectants. Band intensities were quantified using ImageJ, and values are shown next to each band. Positive results are highlighted in red, and negative results are indicated in black. ( B ) The quantification of band intensities from ( A ), focusing on the groups outlined in black. The strongest signal, “Supernatants-6” from the ASFV-only group (190.24), was set as 100%, and all other values were normalized accordingly. Asterisks indicate statistically significant differences between groups (*** p < 0.001), determined by a one-way ANOVA.

Article Snippet: Cells were blocked with 5% fetal bovine serum (FBS) in PBS for 30 min and incubated overnight at 4 °C with a primary antibody specific to ASFV p54 (1:200 dilution in blocking buffer, GeneTex, Irvine, CA, USA).

Techniques: Lateral Flow Assay

Journal: Pathogens

Article Title: A Practical Framework for ASFV Disinfectant Evaluation: Differentiating Cytopathic Effects from Cytotoxicity via Integrated Analytical Methods

doi: 10.3390/pathogens14050451

Figure Lengend Snippet: Detection of ASFV antigen (p54) and cellular integrity under different treatment conditions. Vero cells were treated with ASFV alone, ASFV combined with disinfectants at dilution rates of 175×, 275×, and 375×, or left untreated (N.C.). Cells were examined by bright-field microscopy, an indirect immunofluorescence assay (IFA; anti-p54, green), and DAPI staining (blue). The ASFV antigen was detected only in the ASFV-only group. Nuclear damage increased with higher disinfectant concentrations, while the 375× dilution group showed improved nuclear preservation. All images were acquired at the same magnification using an Olympus IX73 inverted microscope equipped with a 20×/0.40 Ph1 objective (total magnification: 200×). Images were cropped for layout consistency.

Article Snippet: Cells were blocked with 5% fetal bovine serum (FBS) in PBS for 30 min and incubated overnight at 4 °C with a primary antibody specific to ASFV p54 (1:200 dilution in blocking buffer, GeneTex, Irvine, CA, USA).

Techniques: Microscopy, Immunofluorescence, Staining, Preserving, Inverted Microscopy

Journal: Pathogens

Article Title: A Practical Framework for ASFV Disinfectant Evaluation: Differentiating Cytopathic Effects from Cytotoxicity via Integrated Analytical Methods

doi: 10.3390/pathogens14050451

Figure Lengend Snippet: LDH release and MTT assay-based cytotoxicity analysis across experimental groups. ( A ) Daily LDH release (%) measured over six days (DPI 1–6) to assess cytotoxicity. ( B ) MTT assay absorbance values on DPI-6 to evaluate cell viability. ( C ) Cumulative LDH-based cytotoxicity (%) across the experimental period. ( D ) MTT assay-based cell viability (%) on DPI-6. Statistically significant differences were observed between the 375× + ASFV group and the D 375× group ( p < 0.01), as determined by a one-way ANOVA followed by post hoc analysis. Asterisks (**) indicate p < 0.01.

Article Snippet: Cells were blocked with 5% fetal bovine serum (FBS) in PBS for 30 min and incubated overnight at 4 °C with a primary antibody specific to ASFV p54 (1:200 dilution in blocking buffer, GeneTex, Irvine, CA, USA).

Techniques: MTT Assay

Journal: Frontiers in Immunology

Article Title: Vaccination With a Gamma Irradiation-Inactivated African Swine Fever Virus Is Safe But Does Not Protect Against a Challenge

doi: 10.3389/fimmu.2022.832264

Figure Lengend Snippet: Electron micrograph of a 30 kGy irradiated African swine fever virus shows the integrity of the particle. Negative staining with 1% phosphotungstic acid. Scale bar 100 nm.

Article Snippet: To verify the integrity of the p72 antigen in the irradiated virus suspension, the Ingezim ® ASFV CROM Ag (Eurofins Technologies Ingenasa) lateral flow assay, which is a double antibody sandwich immunochromatographic assay for the detection of ASFV antigen in blood samples ( ) was used.

Techniques: Irradiation, Negative Staining