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.

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.

Chlorine dioxide (ClO2) has recently emerged as an ideal disinfectant and has shown a wide range of antimicrobial activities in various pathogenic microorganisms. In this study, the virucidal effect of ClO2 at low concentration (0.02 ppm) and higher concentration (0.06 – 0.09 ppm) against Adenovirus and Herpesvirus was evaluated based on the NF T 72-281 and ASTM 1053-11 standard methods at different exposure times. The virus suspension was dried onto the carrier and then exposed to gaseous ClO2 (gClO2) at 22 ± 2∘C. For Adenovirus, exposure at a low concentration of ClO2 at the middle height resulted in the average log10 reduction of 0.95, 2.65, and 5.30 after 1, 3, and 6 h post-exposure (pe), respectively. Moreover, more than 4-log10 reduction was achieved at 4 and 6 h pe with higher concentrations of ClO2. On the other hand, the antiviral activity of gClO2 at the middle height was also effective against Herpesvirus. In particular, at 1 h pe, a less than 4-log10 reduction was observed at all examined concentrations of ClO2, whereas exposure for 3 and 6 h (with low concentration) or 2 h (with higher concentration) inactivated completely viruses attached to the carrier. These results suggested that ClO2 fumigation is a potential alternative method for disinfecting healthcare facilities, high-containment laboratories, and households with a safe concentration for human health.

The mechanical safety of the container designed according to the IP-2 type technology standard was analyzed for the temporary storage and transportation of Very-Low-Level-Waste (VLLW) for liquid occurring at the nuclear facilities decommissioning site. The container was designed and manufactured as a composite shielding container with the effect of storing and shielding liquid radioactive waste using High Density Polyethylene (HDPE) and eco-friendly shielding material (BaSO4) with corrosion and chemical resistance. The main material of the composite shielding container is HDPE and BaSO4, the material of the cover, cage and pallet is SUS304, and the angle guard is elastic rubber. The test and analysis requirements were analyzed for structural analysis of container drop and lamination test. As test requirements for IP-2 type transport containers should be verified by performing drop and lamination tests. There should be no loss or dispersion of contents through the 1.2 m high free-fall drop and lamination test for a load five times the amount of transported material. ABAQUS/Explicit, a commercial finite element analysis program, was used for structural analysis of the drop and lamination test of the transport and storage container. (Drop test) It was confirmed that the container was most affected when it falls from a 45-degree slope. Although plastic deformation was observed at the edge axis of the cover, it was evaluated that the range of plastic deformation was limited to the cover and cage, and stress within the elastic limit occurred in the inner container. In the analysis results for other falling direction conditions, it was evaluated that stress within the elastic limit was generated in the inner container except for minor plastic deformation. In the case of on-site simulation evaluation, deformation of the inner container and frame due to the drop impact occurred, but leakage and loss of contents, which are major evaluation indicators, did not occur. (Lamination test) The maximum stress was calculated to be 19.9 MPa under the lamination condition for a load 5 times the container weight, and the maximum stress point appeared at the corner axis of the pallet. The calculated value for the maximum stress is about 10%, assuming the conservative yield strength of SUS304 is 200 MPa. It was evaluated that stress within the limit occurred. In the case of on-site simulation evaluation, it was confirmed that there was no container deformation or loss of contents due to the load.

We investigated the effect of a synthetic complement peptide C3a on the outcome of Brucella abortus 544 infection in a murine macrophage cell line RAW264.7 cell. First, we determined the highest non-cytotoxic concentration of the peptide in the cell line. We also found that the peptide significantly increased the growth of the bacteria at 8 and 24 h. Although the number of bacterial CFU was also elevated at 48 and 72 h, the increases were not significant as compared to controls. We further investigated the effect of C3a peptide on the growth of Brucella by pre-incubating the peptide at various temperatures and found that the effect was reversed at 24 h post-incubation suggesting that incubation of peptide at high temperatures including 65°C or 95°C could inactivate its action. This also could indicate the beneficial effect of high temperature during infection. Although several studies reported the inhibitory effect of different antimicrobial peptides including C3a, the present study preliminarily revealed that it had no positive contribution on the control of B. abortus 544 infection in vitro and indirectly to its receptor, CD88, which belongs to GPCR. Moreover, the encouraged further exploration of the effect of other similar peptides would be performed for the purpose of finding Brucella-host cell interaction for the control of disease progression.

This study aims to investigate the effects of exogenous succinic acid (SCA) on Brucella (B.) abortus infection in macrophage RAW 264.7 cells and ICR mice. Firstly, the in vitro experiment was conducted by MTT cytotoxicity and bacterial internalization assay to evaluate the uptake of B. abortus into macrophage cells. Two non-cytotoxic concentrations of SCA demonstrated attenuated invasion of Brucella into macrophages at 30 and 45 min post- infection (pi). Secondly, ICR mice were treated with SCA and infected with B. abortus. On day-14 pi, spleen and blood serum were collected to evaluate the bacterial burden and total spleen weight as well as the production of cytokine/chemokine, respectively. The results showed that SCA treatment promoted bacterial growth and reduced the total spleen weight in mice. Furthermore, SCA treatment increased the level of IL-10 cytokine in the sera, while dampening the production of MCP-1 chemokine compared to the control. The results of bacterial load in spleen and spleen weight together with cytokine/chemokine production profile in the sera indicated that SCA induced the host anti-inflammatory response which is beneficial for the survival of Brucella. Therefore, these findings suggest that SCA contributed to host immunity against Brucella infection and the emerging potential topic-immunometabolism should be invested for further investigations.

We investigated the cytotoxic potential of three different commercially available absorbent feminine hygiene products and one transdermal patch using direct contact and extract exposure methods. Two different cell lines were used – mouse fibroblast L929 and normal human skin fibroblast CCD-986sk cell lines. The test samples were extracted using three different methods in accordance to International Organization for Standardization (ISO). Viability of cells was analyzed using MTT assay and morphology of the cells were also observed using phase contrast microscopy. Overall, the direct contact method using L929 cells showed that all the test samples had no toxic effect when exposed to extracts for 1 h. For the exposure method, no toxic effect was observed in both L929 and CCD986sk cells incubated with all the test samples regardless of the extraction methods used.

This study investigated the efficacy of four Brucella (B.) abortus recombinant proteins, namely adenylate kinase (Adk), nucleoside diphosphate kinase (Ndk), 50S ribosomal protein (L7/L12) and preprotein translocase subunit (SecB), as a combined subunit vaccine (CSV) against B. abortus infection in BALB/c mice. Immunoblotting assay showed that these four recombinant proteins as well as pcold-TF vector reacted individually with Brucella-positive serum, but not with Brucella-negative serum. The peripheral blood CD4+ T cell population was increased in CSV-immunized mice compared to PBS and pcold-TF vector groups. In addition, CSV and pcold-TF groups displayed induced IgG1 and IgG2a antibodies production compared to PBS and RB51 group, whereas IgG2a titer was higher than IgG1 titer in CSV group. The secretion profiles of IgG1 and IgG2a production together with an enhancement of CD4+ T cell population suggested that CSV did not only induce T helper 1 (Th1) T cell immunity but also humoral immunity. Therein, Th1 T cell immunity is more predominant in eliminating intracellular bacteria B. abortus. Furthermore, CSV immunization significantly reduced the bacterial burden in the spleen as well as the spleen weight in comparison to PBS and pcold-TF groups. Altogether, combination of these antigens could be potential to induce protective immunity against B. abortus infection in animals.

BALB/c mice were vaccinated with Brucella (B.) abortus recombinant protein L27 (50S ribosomal protein L27) cloned into a pMal vector system. L27 was induced, purified and injected intraperitoneally (IP). Mice were vaccinated on 0-, 15- and 35-day. Serum cytokines were evaluated on 36- and 49-day from first vaccination. Mice were intraperitoneally infected with 5×104 CFU of virulent B. abortus 544 on day-50 and sacrificed after two weeks from infection. Bacterial burden from the spleen was quantified and showed a 0.7- and 0.9-log reduction in vaccinated mice in comparison to PBS and MBP (maltose binding protein) groups respectively. Cytokines in the serum demonstrated increased interferon-gamma (IFN-γ) and other pro-inflammatory cytokines such as macrophage chemoattractant protein-1 (MCP-1) and interleukin 6 (IL-6). On the other hand, interleukin 10 (IL-10) was attenuated in the sera of vaccinated mice. This cytokine profile is indicative of a cell-mediated type of immune response which is favorable for the eradication of intracellular infections. The current study showed the potential of another B. abortus ribosomal protein in inducing protective immunity against B. abortus infection.

In this study, we examined the protective immunity of a combination of seven Brucella abortus recombinant proteins; superoxide dismutase (rSodC), riboflavin synthase subunit beta (rRibH), 50S ribosomal protein (50s rL7/L12), nucleoside diphosphate kinase (rNdk), malate dehydrogenase (rMDH), arginase (rRocF), and elongation factor (rTsf) cloned in a pMal vector system and expressed in DH5α. Mice groups were immunized thrice with a combined subunit vaccine (CSV-7) at 0, 2, and 5 weeks and subsequently challenged with B. abortus at 5 × 104 CFU at 6 weeks. At four weeks post-infection, the mice were sacrificed and the bacterial burden in their spleens was quantified. Results revealed bacterial log reductions of 0.63 and 0.34 in comparison to PBS and maltose-binding protein (MBP), respectively. Cytokine profiling revealed a marked increase in IFN-γ (interferon-gamma), MCP-1 (macrophage chemoattractant protein-1) and IL-6 (interleukin 6) cytokines at 5-weeks post-immunization. On the other hand, only TNF was heightened at 7-weeks post-immunization. In general, this cytokine profile is consistently reflective of a Th1 immune response, which is beneficial for host immunoresistance.

This study evaluated the protective effects of a combination of eight B. abortus recombinant proteins that were cloned and expressed into a pMal vector system and DH5α: nucleoside diphosphate kinase (rNdk), 50S ribosomal protein (rL7/L12), malate dehydrogenase (rMDH), DNA starvation/stationary phase protection protein (rDps), elongation factor (rTsf), arginase (rRocF), superoxide dismutase (rSodC), and riboflavin synthase subunit beta (rRibH). The proteins were induced, purified, and administered intraperitoneally into BALB/c mice. The mice were immunized three times at weeks 0, 2, and 5 and then infected intraperitoneally (IP) with 5×104 CFU of virulent B. abortus 544 one week after the last immunization. The spleens were collected and the bacterial burden was evaluated at four weeks post-infection. The results showed that this combination produced a significant reduction of the bacterial burden in the spleen with a log reduction of 1.01 compared to the PBS group. Cytokine analysis revealed induction of the cell-mediated immune response in that TNF (tumor necrosis factor) and proinflammatory cytokines IL-6 (Interleukin 6) and MCP-1 (macrophage chemoattractant protein-1) were elevated significantly. In summary, vaccination with a combination of eight different proteins induced a significant protective effect indicative of a cell mediated immune response.

The present study was carried out to investigate the acute oral toxicity of Chamaecyparis obtusa (C. obtusa) essential oil in ICR male and female mice. Acute oral treatment with C. obtusa essential oil did not reveal any sign of toxicity or mortality in treated mice. Mouse body weights were not affected after single oral administration of C. obtusa essential oil during the 14-day observation period. In the hematological and blood biochemical analysis, all parameters of the treated group with 2,000 mg/kg body weight of the essential oil were not significantly different those of the control group. Therefore, the lethal dose 50 of the essential oil was estimated to be greater than 2,000 mg/ kg body weight in mice, which indicated that the essential oil is non-toxic. In conclusion, this study suggests that C. obtusa essential oil orally safe ICR mice.

본 연구는 정향추출물의 E. coli O157:H7에 대한 항균 효과와 E. coli O157:H7 감염 마우스에 대한 치료효과를 평가하기 위해 수행되었다. 정향 메탄올 추출물(FSAE)을 이용하여 E. coli O157:H7 에 대한 항균효과 확인 시험을 수행한 결과, 배양 후 24 시간째에, FSAE를 첨가한 모든 군들에서 E. coli O157:H7 의 생존율이 무투여 대조군에 비해 통계적으로 유의성 있게 감소하는 결과를 보여(0.269 mg/ml, p < 0.05; 0.538과 1.075 mg/ml, p < 0.001), FSAE가 E. coli O157:H7의 증식 억제 효과가 뛰어난 것으로 확인되었다. 또한, E. coli O157:H7을 감염시킨 마우스에 FSAE를 경구로 투여한 결과, 투여 후 3일째에, FSAE를 1.075 (p < 0.05)와 2.15 mg/ ml (p < 0.01)로 투여한 군들에서 대조군과 비교하여 분변 내 E. coli O157:H7의 균수가 유의성 있게 감소하였으며, 투여 7일째에는, 모든 FSAE 투여군들에서 대조군과 비교 하여 E. coli O157:H7의 균수가 통계적으로 유의성 있게 감소하였다(0.538 mg/ml, p < 0.05; 1.075와 2.15 mg/ml, p < 0.001). 이상의 연구결과로부터, FSAE를 E. coli O157: H7에 감염된 마우스에 경구로 투여할 경우, 감염증상을 완화 시킬 수 있을 것으로 기대된다.

Antibiotic Detection Kit (Combination I), a lateral flow immunoassay (LFIA) developed for detecting antibiotic residues in milk, was utilized for the analysis of antibiotic residues in the muscle tissue of olive flounder. After 5-h treatment of samples by placing them in water dosed with sulfadimethoxine (SDM; 200 g/ton water), the residue depletion of SDM was investigated in 25 cultured olive flounders (Paralichthys olivaceus). Muscles from fish were sampled before treatment and on the 1st, 2nd, 3rd, 4th and 5th days after treatment. The concentration of SDM in the muscle was then determined by LFIA. The absorbance ratio of the sample to the control blank (Bs/Bo) was employed as an index to determine the residue in olive flounder muscle. To investigate the recovery rate, standard solutions were added to muscle samples to obtain final concentrations of 25 and 50 ng/mL in the muscle. The recovery rates of all spiked samples were >96.6% of the spiked value. SDM was detectable in the muscle of fish treated with the drug until the 1st day of the withdrawal period. The present study shows that the LFIA can be easily adopted to detect SDM residues in the tissue of farmed fish.

This study investigated the antibacterial effects of Galla rhois extract (GRE) against Campylobacter jejuni and Campylobacter coli. The minimum inhibitory concentrations (MICs) of GRE against C. jejuni and C. coli were 0.28 and 0.55 mg/mL, respectively, and the corresponding minimum bactericidal concentrations (MBCs) were 4.4 and 5.5 mg/mL. C. jejuni treated with the MIC, MBC or 2×MBC of GRE showed significant inhibition of growth compared with that of the control group during the incubation period, and no viable bacteria were detected at 24 h after incubation. C. coli treated with MIC, MBC or 2×MBC of GRE also showed inhibition of growth compared with that of the control group during the incubation period, and in the C. coli cultures treated with MBC and 2×MBC of GRE, no viable bacteria were detected at 24 h after incubation. In conclusion, GRE is a candidate antibacterial agent against C. jejuni and C. coli, and may have applications for the control of Campylobacter infection in poultry.

To date, most serodiagnostic methods for brucellosis screening are based on antibodies against lipopolysaccharides of Brucella spp. However, this approach has the drawback of yielding false-positive results due to cross-reactivity with lipopolysaccharides of other related pathogens, especially Yersinia enterocolitica O:9. In this study, Brucella abortus AspC was cloned and expressed by PCR amplification into a pCold TF expression system to obtain recombinant AspC (rAspC). The immunogenicity of rAspC was confirmed by western blotting of Brucella-positive bovine serum. rAspC-based ELISA was performed to determine whether rAspC could be used in the serodiagnosis of bovine brucellosis. rAspC reacted strongly with anti-Brucella antibodies in positive sera in the tube agglutination test (TAT), but did not show strong reaction with most negative samples. In particular, the average OD492 value at the highest TAT titer showed a 1.4-fold increase with respect to the cutoff value. The accuracy, specificity, and sensitivity of rAspC were 71.88%, 78.33%, and 68%, respectively. These findings suggest that rAspC might be valuable for the serological diagnosis of bovine brucellosis.