Porcine edema disease (ED) is an enterotoxemia of pigs caused by Escherichia coli that produces Stx2e. In this study, the protective efficacy of a recombinant modified Stx2e toxoid was evaluated as a vaccine candidate against ED in piglets. The recombinant Stx2e toxoid was expressed and purified using a commercial E. coli expression system. A total of 25 piglets were used and divided into 5 groups (A to E), with 5 piglets in each group. All piglets (except those in group A) were intramuscularly immunized at 5 days of age (0 weeks post prime immunization; 0 WPPI) and again at 3 weeks of age (2 WPPI). Group B piglets were inoculated with sterile PBS, while groups C to E were immunized with 25 μg/piglet, 50 μg/piglet, and 100 μg/piglet of the recombinant toxoid, respectively. All piglets in groups B to E were orally challenged with virulent wild-type Stx2e⁺ F18⁺ E. coli isolates at 5 weeks of age (4 WPPI). Serum IgG titers in groups D and E were significantly increased from 2 WPPI until the end of the study. Furthermore, no clinical signs were observed in groups A and E during the 7 days following the challenge, while clinical signs of ED were observed in 80%, 60%, and 20% of piglets in groups B, C, and D, respectively. These results indicate that intramuscular vaccination with 100 μg/piglet of the recombinant modified Stx2e toxoid can provide effective protection against ED in piglets.

The primary therapeutic approach for Brucella species infections has mainly been based on antibiotic treatment. However, the development of vaccines for brucellosis control remains controversial. Furthermore, there is currently no licensed vaccine available for human brucellosis. This study aims to evaluate the effect of a combination of recombinant protein vaccines against Brucella (B.) abortus infection using a mouse model. Two B. abortus genes, namely dapB and gpm, were cloned and expressed in competent Escherichia (E.) coli DH5α using the pCold-TF vector. Successfully cloned vectors were subjected to PCR amplification using specific primer pairs. The apparent sizes of dapB and gpm were detected at 807 bp and 621 bp, respectively. Besides, the purified recombinant proteins dapB and gpm were detected using SDS-PAGE electrophoresis with correct sizes of 82.86 kDa and 87.61 kDa, respectively. These recombinant proteins were used to immunize mice as a combined subunit vaccine (CSV) to elicit host immunity against B. abortus infection. Mice immunized with CSV exhibited increased proliferation of CD4+ and/or CD8+ T cells at week 7th and 9th before sacrifice, in comparison to the control group. Notably, CSV immunization showed a significant decrease in bacterial burden in the spleen compared to the control group. Altogether, CSV using dapB and gpm induced host adaptive immune response against Brucella infection, suggesting its potential as an effective new subunit vaccine candidate.

To identify some significant phenotypic characteristics of maize(zea mays) seeds, we have obtained Red, Green, Blue(RGB) digital image data from 82 recombinant inbred lines. Based on the collected image data, their morphological and color data were analyzed, and seven significant parameters were selected, including area, perimeter, length, width, circularity, roundness, and surface texture. The extracted RGB data were converted into color hex codes to visualize the representative colors of the seeds. These visualized colors were categorized into six groups: gray, yellowish white, yellow, grayish orange, purple, and brown. The results of maize seed phenotypic analysis using the RGB digital images in this study will serve as a useful tool for constructing a database of seed phenotyping database and establishing a standardized classification system.

Extensive research and testing continue to be conducted for the development of vaccines targeting zoonotic diseases such as brucellosis. In this study, the potential of the DapB as a recombinant protein vaccine to effectively combat Brucella abortus 544 infection in BALB/c mice was evaluated. Western blotting assay results showed that recombinant protein DapB reacted with Brucella-positive serum, indicating its potential immunoreactivity. In vivo results showed that the peripheral blood CD4+ and CD8+ T cell population significantly increased in the DapB-immunized mice group after the first, second and third blood collection, compared to the control group that received PBS. Additionally, at the fourth blood collection, an increase in CD4+ T cell activation was observed in three vaccination groups compared to PBS negative control group. These results indicate the potential of DapB in stimulating cellular immunity. Fourteen days after infection, the bacterial load in the spleen was evaluated. The reduction in bacterial replication in the spleen by both DapB and RB51 highlights their protective efficacy against Brucella infection. These findings contribute to the ongoing efforts in developing effective vaccines against brucellosis and provide valuable insights for further research in this field.

Subunit vaccines are being developed as a potential therapy for preventing microbial pathogen infection. In this study, the immunogenicity of recombinant Brucella (B.) abortus Fe/Mn superoxide dismutase (rFe/Mn SOD) protein as a subunit vaccine against B. abortus was investigated in BALB/c mice model. Brucella Fe/Mn SOD gene was cloned into a pcold-TF DNA vector. The bacterial recombinant protein was expressed using the Escherichia coli DH5α strain with a size of 82.50 kDa. The western blotting assay showed that rFe/Mn SOD reacted with Brucella-positive serum, indicating the potential immunoreactivity of this recombinant protein. After the second and third vaccinations, the peripheral CD4+ T cell population was increased significantly in the rFe/Mn SOD-immunized mice group compared to the PBS control group. Moreover, immunization of this recombinant protein increased the CD4+ T cell population from the first vaccination to the third vaccination. Meanwhile, the CD8+ T cells were slightly enhanced after the second vaccination compared to the first vaccination and compared to control groups. Fourteen days after the bacterial infection, the splenomegaly and the number of bacteria in the spleen were evaluated. The result showed that both rFe/Mn SOD and positive control RB51 decreased the bacterial replication in the spleen and the splenomegaly compared to control groups. Altogether, these results suggested that rFe/Mn SOD could induce host immunity against B. abortus infection.