Effects of different stocking density and various phytogenic feed additives dosage levels on growing-finishing pigs.

This study was to investigate the effects of different phytogenic feed additives (PFA) dosage levels in growing- finishing pigs stressed by high stocking density. A total of 72 mix sexed 12 weeks growing pigs ([Landrace × Yorkshire] × Duroc) with initial body weight (BW) of 49.28 ± 4.58 kg were used for 8 weeks. There were 3 replicate pens in each treatment group, with 3 pigs per pen. The dietary treatment groups consisted of basal diets in animal welfare density (negative control [NC]), basal diet in high stocking density (positive control [PC]), PC + 0.04% essential oil (ES1), PC + 0.08% essential oil (ES2), PC + 0.10% bitter citrus extract & essential oil (CES1), PC + 0.20% bitter citrus extract & essential oil (CES2), PC + 0.05% grape pomace extract (GP1), PC + 0.10% grape pomace extract (GP2). The reduction of space allowance decreased (p < 0.05) average daily gain, feed efficiency, and digestibility of dry matter, crude protein, and gross energy. Also, the fecal score of PC groups increased (p < 0.05) compared with other groups. Basic behaviors (feed intake, standing, lying) were inactive (p < 0.05) and singularity behavior (biting) was increased (p < 0.10) under high stocking density. There was no difference in blood profile. However, the supplementation of PFA alleviated the negative effects such as reduced growth performance, nutrient digestibility, and some increasing stress indicators in th blood (cortisol) and animal behavior (biting). In conclusion, the negative effect of high stocking density was most effectively mitigated by the normal dosage of the mixture of bitter citrus extract and essential oil additive (CES1).

Effect of insect protein and protease on growth performance, blood profiles, fecal microflora and gas emission in growing pig.

Two experiments were conducted to determine the effect of Hermetia illucens larvae (HIL) as protein and protease on growth performance, blood profiles, fecal microflora, and gas emission in growing pig. In experiment 1, the seventy-two crossbred growing pigs ([Landrace × Yorkshire] × Duroc) with an initial body weight (BW) of 27.98 ± 2.95 kg were randomly allotted to one of four dietary treatments (3 pigs per pen and 6 replicates pen per treatments). The experimental design was a 2 × 2 factorial arrangement of treatments evaluating two diets (Poultry offal diets and HIL diets) without or with supplementing protease. The poultry offal in basal diet has been replaced by HIL. In experiment 2, the four crossbred growing pigs ([Landrace × Yorkshire] × Duroc) with an initial BW of 28.2 ± 0.1 kg were individually accepted in stainless steel metabolism cages. The dietary treatments included: 1) PO- (PO-; poultry offal diet), 2) PO+ (PO- + 0.05% protease), 3) HIL- (3% PO of PO- diet was replacement 3% HIL), 4) HIL+ (HIL- + 0.05% protease). In experiment 1, From weeks 0 to 2, average daily gain (ADG) and feed efficiency (G:F) were significantly increased in the PO diet group compared with the HIL group. From weeks 2 to 4, ADG and G:F were higher for protease group than for non-protease group. At weeks 2 and 4, the PO diet group had lower blood urea nitrogen (BUN) levels than HIL diet group. In experiment 2, crude protein (CP) and nitrogen (N) retention were decreased by HIL diet at weeks 2 and 4. The fecal microflora and gas emission were not affected by HIL and protease. The HIL diet showed lower CP digestibility than PO diet and total essential amino acids digestibility tended to higher in PO diet than HIL diet. In summary, the present study revealed that replacement of the PO protein with the HIL protein and the additive of protease in growing pig diets during the overall experimental period had no negative effect.

Effects of tumor vaccine expressing Granulocyte-Macrophage Colony Stimulating Factor and interleukin-18 fusion on cancer cells and its possible application for cancer immunotherapy.

To access antitumor effects of a combined Granulocyte Macrophage-Colony Stimulating Factor (GM-CSF) and interleukin-18 (IL-18), cDNA fusion of murine GM-CSF and mature IL-18 (GMIL-18) was constructed and transfected in mammalian cells. GMIL-18 fusion protein was highly secreted and displayed bifunctional activities, possessing immune response initiation and cytokine roles, including IFN-γ induction in mouse splenocytes and increased proliferation of GM-CSF-dependent cells, M-NSF-60. The GMIL-18 secreting tumor vaccine was generated and it strongly stimulated differentiation of dendrite cells (DCs) and effusive CD8+ and CD4+ cell infiltration into tumor mice. Moreover, growth of CT26 mouse colon cancer cells was significantly retarded by GMIL-18 (CT26GMIL-18), but not by CT26GM-CSF- or CT26IL-18. The efficiency of prophylactic vaccination was greater than that of therapeutic vaccination in terms of tumor size and its inhibitory role in proliferation. In micrometastasis analysis of tumor models, γ-ray irradiated GMIL-18 tumor vaccine showed a smaller number of liver-meta tumor nodules in mouse liver cells. We concluded that bifunctional GMIL-18 fusion protein could be applied as an immune therapy for cancer treatments.

Neuroprotective activity of Viola mandshurica extracts on hydrogen peroxide-induced DNA damage and cell death in PC12 cells.

This study was conducted to examine the neuroprotective effects of acetone extracts from Viola mandshurica (VME). The effect of VME on hydrogen peroxide (H(2)O(2))-induced DNA damage in PC12 cells was evaluated by the comet assay where VME (100 and 250 microg/mL) was a dose-dependent inhibitor of DNA damage induced by 500 micromol/L of H(2)O(2). The protective effect of VME against H(2)O(2)-induced oxidative damage on PC12 cells was investigated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] reduction assay and lactate dehydrogenase (LDH) release assays. After 3 h of cell exposure to 500 micromol/L of H(2)O(2), a marked reduction in cell survival was observed. However, the reduction was significantly prevented by 100 and 250 microg/mL of VME. H(2)O(2) also induced severe apoptosis of the PC12 cells, which was indicated by Hoechst 33342 staining. Interestingly, the H(2)O(2)-stressed PC12 cells that were incubated with 100 and 250 microg/mL of VME had greatly suppressed apoptosis. The results suggest that VME could be a new antioxidant candidate against neuronal diseases.

Comparison of conventional, nested, and real-time PCR assays for rapid and accurate detection of Vibrio vulnificus.

We conducted a prospective study to target toxR in the blood of patients with skin and soft tissue infections who were admitted to four tertiary hospitals to assess the clinical usefulness of real-time quantitative PCR (Q-PCR) as a diagnostic technique. We performed conventional PCR (C-PCR), nested PCR (N-PCR), and Q-PCR assays and compared the results to those obtained using the "gold standard" of microbiological culture. The lower detection limit for the Q-PCR assay was 5 x 10(0) copies/microl. By use of blood samples of patients with skin and soft tissue infections, the sensitivities of the C-PCR and N-PCR assays against the target toxR gene of V. vulnificus as diagnostic tools were determined to be 45% and 86%, respectively. The C-PCR and N-PCR assays had specificities of 100% and 73%, respectively. When we adopted a crossing-point (cp) cutoff value of <38 cp as a positive result, the Q-PCR assay had 100% sensitivity and specificity. Q-PCR to detect V. vulnificus-specific genes is not only the most sensitive and specific of the techniques but also the most rapid diagnostic method. Therefore, the appropriate application of the Q-PCR assay using blood is useful for the rapid diagnosis and subsequent treatment of V. vulnificus sepsis.

Overexpression of extracellular superoxide dismutase (EC-SOD) in mouse skin plays a protective role in DMBA/TPA-induced tumor formation.

Extracellular superoxide dismutase (EC-SOD, EC 1.15.1.1) is a major antioxidant enzyme that is located in the extracellular matrix and on the cell surface. EC-SOD protects against cell and tissue damage initiated by extracellular-produced reactive oxygen species (ROS). We investigated a major role of EC-SOD in the development of tumor formation. In this study, we reported that skin-specific overexpressed EC-SOD transgenic mice showed half the number of tumors compared with the nontransgenic mice in the dimethylbenzanthracene (DMBA)-initiated and a 12-O-tetradecanoylphorbol-13-acetate (TPA)-promoted two-stage skin carcinogenesis model. This model showed a significant increase of the epidermal cell proliferation in the nontransgenic mice, but the proliferative response in the transgenic mice was delayed. The 8-hydroxy-2'-deoxyguanosine (8OH-dG) detection assay showed that the oxidative DNA damage was significantly higher in the nontransgenic mice than in the transgenic mice after TPA treatments. Overall, EC-SOD overexpression inhibited the TPA-induced cell proliferation and DNA damage, and reduced the subsequent formation of tumors. Our data suggest that EC-SOD plays a protective role in DMBA/TPA-induced skin carcinogenesis.