The use of non-therapeutic antibiotics as animal feed additives has raised public health concerns due to the increasing resistance of pathogens to antibiotics. It is therefore required to develop safe and effective alternative feed additives to replace non-therapeutic antibiotics. The aim of this study was to assess the effects of the multiherbal compound, KIOM-C, on growth performance and immune response of growing-to-finishing pigs under farm conditions. The experimental trials were performed in a Korean commercial swine growing-to-finishing complex, and a total of 70-day-old 160 pigs were selected. Eighty pigs were treated with KIOM-C at the level of 2 kg/tonne until slaughter age (KT group), while another 80 pigs were not treated with KIOM-C (NT group). All animals were vaccinated against foot-and-mouth disease (FMD) at 60 and 110 days of age. During the trial period, average daily weight gain (ADWG), average daily feed intake (ADFI), feed conversion ratio (FCR), survival rates, and average slaughter ages were measured. The serum concentrations of tumor necrosis factor-a (TNF-α), interferon-γ (IFN-γ), and IgA were also evaluated. In order to evaluate specific humoral immune responses, the foot-and-mouth disease virus (FMDV) serotype O-specific antibody was measured. The ADWG, ADFI, and FCR of the KT group were significantly greater than those of the NT group (p<0.05). Serum concentrations of IgA in the KT group was statistically higher than the NT group. The antibody levels of the KT group against FMDV serotype O was higher than the NT group, and 86.67% of the KT group tested positive for anti-FMDV antibodies. Overall, these findings suggest that KIOM-C improves growth performance and immune response of pigs under growing-to-finishing farm conditions, and implies that the herbal compound may be used as a suitable alternative feed additive.

The purpose of this study was to investigate probiotic characteristics and immune enhancing effects of Bifidobacterium (B.) longum KBB1-26 and BIF-4, B. breve KBB5-22 isolated from human intestine for probiotic use in humans and animals. We measured acid, bile and heat tolerance, antimicrobial activity against pathogenic bacteria, Escherichia (E.) coli, Salmonella (S.) Enteritidis, Staphylococcus (S.) aureus, and Listeria (L.) monocytogenes. Immune enhancing effects of B. longum and B. breve were investigated by measuring nitric oxide (NO), nuclear factor (NF–κb), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), interleukin-12 (IL-12) and tumor necrosis factor- α (TNF- α) in RAW 264.7 cells or RAW BLUE cells. B. longum KBB1-26 was survived at pH 2.0. B. longum KBB1-26 and BIF-4, B. breve KBB5-22 also showed tolerance to 0.3% of oxgall bile salt. B. longum KBB1-26 was able to survive at 70 ℃ and 80 ℃ for 20 min. KBB1-26 showed the antimicrobial inhibition zone to pathogenic bacteria such as E. coli (12 mm), S. Enteritidis (14 mm), S. aureus (14 mm) and L. monocytogenes (41 mm). The production of NO (4.5 ± 0.00μM/mL) and IL-1β (39.7 ± 0.55pg/mL) of KBB1-26 significantly higher than BIF-4 and KBB5-22, respectively. In addition, KBB1-26 and KBB5-22 induce the production of high level of TNF-α and IL-6 in macrophages. Collectively, B. longum KBB1-26 have acid, bile, heat tolerance, antimicrobial activity and immune enhancing effects. These results suggest that KBB1-26 can be used as probiotics for humans and animals.

본 연구는 가죽나무(Ailantias altissima) 추출물을 유효성분으로 함유하는 면역 증강용 소재를 개발하기 위하여 수행한 것으로, 가죽나무 에탄올 추출물 단독 또는 면역능을 보유한 유산균과의 혼합처리에 의한 면역 활성능을 비교 분석하였다. 가죽나무 에탄올 추출물에 의한 대식세포 증식에 미치는 영향을 살펴본 결과, 10~500 μg/mL 농도 범위로 처리하였을 때 100 μg/mL 농도에서만 대식세포가 활성화되어 높은 세포 증식률을 나타내었다. NF-κB 활성 평가에서는 유산균과 가죽나무 추출물을 1:0.25 비율로 혼합해서 처리하였을 때 추출물을 단독 처리하였을 때보다 NF-κB 활성능이 증가하였는데, 생균보다 사균과 함께 처리하였을 때 그 값이 더 높았다. 특히 Lactobacillus plantarum 4-25 사균과 추출물을 1:0.25 비율로 혼합 시 가장 높은 활성능을 나타내었다. Nitric oxide(NO) 생성량은 가죽나무 에탄올 추출물의 단독 처리에 의해 5.46±0.06 μM/mL로 LPS 처리 대조군(5.73±0.06μM/mL)과 유사하게 가장 높은 값을 보였지만, 혼합 처리군 에서도 Weissella cibaria JW15와 Lactobacillus rhamnosus GG 등 사균과 가죽나무 추출물을 1:0.25 비율로 처리하였을 때만 대조군보다 높은 생성량을 보였다. TNF-α와 IL-1β 등 사이토카인 생성량에서는 가죽나무 추출물 단독처리군보다 유산균들과의 혼합처리군에서 모두 높은 값을 나타내었는데, 특히 Weissella cibaria JW15 균주를 추출물과 혼합한 경우에만 LPS 처리군보다 높은 TNF-α 활성을 보였고, 생균과 사균 간의 큰 차이 없이 1:0.25 비율로 혼합 시 가장 높은 생성량을 보였다. IL-1β 생성량은 Lactobacillus rhamnosus GG 사균과의 혼합에서는 1:0.25 비율로 처리하였을 때 높은 값을 나타내었고, 가죽나무 추출물 단독 처리와 유사한 수준을 나타내었지만, 나머지 혼합군에서는 IL-1β가 검출되지 않았다. 종합적으로 살펴보면 가죽나무 에탄올 추출물과 유산균주와의 혼합에 의한 면역 활성능을 확인한 결과, 가죽나무 추출물 단독처리보다 유산균과 가죽나무 추출물을 1:0.25 비율로 처리한 혼합물에 의해 높은 면역활성능을 보여, 가죽나무 추출물을 유효성분으로 함유하는 소재를 활용하여 면역 저하증 예방 및 치료용 등 건강기능식품 개발을 위하여 유용하게 이용될 수 있을 것으로 기대된다.

Until now, problems related to shortage of organ for transplantation have been continuing. Pigs are the most suitable animal for xenotransplantation. Although primates are most similar to humans, they are not suitable because they have low productivity. Pigs are more productive than primates, and their organ size and physiological characteristics are similar to humans, with the exception of primates. In this study, we breeding the transgenic minipigs using natural mating to produce transgenic pigs. And, transgenic pigs has transmission rate that follow mendel’s rule. There are 20% hDAF gene, 20% US11 gene and 50% both hDAF and US11 gene in transgenic offsprings. Furthermore, transgenic pigs followed normal litter size, and piglets also has normal sex ratio. To suppress the immune function, experiments were performed using porcine ear fibroblast that transfected with hDAF and US11gene. In Cytotoxicity experiment against human complement, hDAF gene and double transgenic cell with both hDAF and US11 gene showed effect to reduce cytotoxicity rate in all of human complement condition. US11 gene and double transgenic cell were significantly reduce the cytotoxicity ratio in human NK cell. Besides, hDAF gene transgenic cell also reduce immune response in 10:1 concentration of human NK cell. In conclusion, natural mating was efficient method for breeding transgenic pigs. And, hDAF and US11 genes has effect for reduce cytotoxicity against human NK cell and human complement conditions.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.