본 연구는 열 스트레스 (heat stress, HS)에 노출된 브로일러에서 혼합 생균제의 급여가 혈 액 생체지표, 면역반응, 맹장 미생물 및 생산성에 미치는 영향을 조사하였다. 총 400마리의 브로일러 수 컷 (Ross 308)을 각각 100마리씩 4그룹, C (대조군, 실온 25℃), HS (열 스트레스 33℃), HSP (HS플 러스 혼합 생균제 500, 750 mg/kg) 그룹으로 배치하였다. 브로일러의 증체량, 사료섭취, 사료요구율 및 면역기관 무게는 HSP 그룹에서 HS 그룹과 비교했을 때 증가하였다. 혈액 IgG, lymphocytes 농도는 HS 그룹에서 HSP그룹과 비교했을 때 증가하였고, heterophil과 lymphocyte(H:L)비율, 코르티코스테론 농도 및 폐사율은 낮았다. 맹장의 Lactobacillus는 HS 그룹과 비교했을 때 HSP 그룹에서 증가하였으나 Escherichia coli (E. coli), coliform bacteria, aerobic bacteria는 낮아졌다. 본 연구결과는 열 스트레스 에 노출된 브로일러에게 B. subtilis, S. galilaeus 및 Sphingobacteriaceae등 3가지 균주가 포함된 혼합 생균제를 급여해주면 면역반응 증진, 미생물 균형을 유지해줌으로써 폐사율을 낮추고 생산성을 개선할 수 있음을 시사해준다.

본 연구는 열 스트레스 (heat stress, HS)에 노출된 산란계에서 혼합 생균제의 급여가 생산성, 계란품질, 면역반응, 맹장 미생물 및 분 암모니아에 미치는 효과를 조사하였다. 총 400마리의 50주령 Hy-Line brown 산란계를 무작위로 각각 100마리씩 4 그룹, C (대조군, 실온 25℃), HS (열 스트레스 33℃), HSP (HS 플러스 혼합 생균제 500, 750 mg/kg)로 배치하였다. 산란계의 생산성, 계란품질, 비장 무게, 혈액 IgG 및 lymphocyte 농도는 HSP 그룹에서 HS 그룹과 비교했을 때 증가하였고, 코르티코스테론, heterophil과 lymphocyte의 비율 (H:L) 및 폐사율은 유의하게 낮았다. Lactobacillus는 HSP 그룹에서 HS 그룹과 비교했을 때 증가하였으나 Escherichiacoli (E. coli), coliform bacteria 및 aerobic bacteria는 유의하게 낮았다. 분에서 암모니아 발생은 HS 그룹이 HSP 그룹에 비해서 유의하게 증가하였다. 결론적으로, 이러한 혼합 생균제가 여름철 산란계의 더위 피해를 방해주고 면역반응, 맹장 미생물 균형을 경유하여 생산성, 계란품질 및 계분으로부터 악취 발생을 줄이는 데 효율적인 영양전략이 될 수 있음을 나타낸다.

곤충의 면역반응에 대한 연구는 곤충 체내 침입한 미생물들과 직접 반응하는 기작들을 중심으로 연구되었다. 그러나 미생물들이 곤충 체내에 침입 한 후 발생되는 다양한 미생물 분비물질에 의한 곤충 면역반응의 시작여부 등에 대한 연구는 거의 없는 실정이다. 이를 위하여 흰점박이 꽃무지(Protaetia brevitarsis seulensis) 유충의 장내에 존재하는 공생균과 체외 병원균을 동일한 조건에서 배양 하고 다양한 분비물질들이 존재 할 거라 예상되는 배양액만을 분리, 유충에 주사하여 면역반응 여부를 조사하였다. 공생균 배양액을 주입한 유충들은 비교적 건강하고 면역반응도 발생하지 않았으나 병원균 배양액을 주입한 유충의 경우 150시간 후 60% 이상 사망하였고 주사된 자리도 짙은 갈색의 멜라닌화가 관찰 되었다. 이러한 면역반응은 과립혈구세포의 리소좀(Lysosomes) 활성화 여부로 재확인 하였다. 병원균 배양액이 주입된 유충들의 경우 12시간 후 리소좀 이 ~50% 이상 활성화 되었으나 공생균 배양액이 주입된 유충들의 경우 ~5% 미만으로 활성화 되는 것으로 나타났다. 따라서 공생균 배양액내에 는 기주면역반응을 유도하는 물질들이 없거나 량이 매우 적게 존재하는 것을 추측 할 수 있었다.

Viral respiratory infections are common in horses, notably equine herpesvirus infection and equine influenza, which primarily initiate secondary bacterial respiratory infections such as strangles caused by Streptococcus equi equi. A decline in the production of stallions has been associated with these respiratory diseases leading to adverse financial implications. This study investigated the antibody responses against respiratory diseases in horses from Jeju Island a year after vaccination. A low level of equine herpesvirus type 1 (EHV-1) (11.36%) antibodies was detected from stallions, however a high level of EHV-4 (95.84%) antibodies was detected from horses without vaccination against this infection suggesting that EHV-4 is ubiquitous in this horse population. In case of equine influenza, ranch stallions showed low positive rate (12.06%) whereas stallions from Subtropical Livestock Research Institute displayed higher positive rate (81.32%). Antibody responses against equine influenza and strangles revealed positive rates of 26.32% and 55.12%, respectively. These findings may draw attention towards the importance of developing an improved disease prevention and/or immunization program that will effectively control respiratory diseases in horses.

Macrophages play an important role in both the innate and adaptive immune responses. These include phagocytosis, killing of microorganisms, antigen presentation, and induction of immune cytokines and antimicrobial genes. Macrophage activity is reported to be controlled by diverse exogenous antigenic or endogenous metabolic molecules, and the underlying mechanisms are well documented in human and mouse macrophage cells. Bacterial lipopolysaccharide (LPS) is known to be one of the most potent stimuli activating macrophages through the toll like receptor 4 (TLR4) signaling pathway. There are other antigenic molecules, such as muramyl dipeptide (MDP) and outer membrane protein A (OmpA), that are also known to activate immune cells. On the other hand, short chain fatty acids (SCFAs) such as acetate and butyrate are produced by gut microbiota and control host energy metabolism and signal transduction through GPR receptors. However, there are few studies demonstrating the effects of these molecules in macrophages from domestic animals, including domestic pigs. In this study, we attempted to characterize gene expression regulation in porcine macrophages (PoM2, Pig Monocytes clone 2) following treatment with LPS, MDP, OmpA, and two short chain fatty acids using porcine genome microarray and RT-PCR techniques. A number of novel porcine genes, including anti-microbial peptides and others, appeared to be regulated at the transcriptional level. Our study reports novel biomarkers such as SLC37A2, TMEN184C, and LEAP2 that are involved in the porcine immune response to bacterial antigen LPS and two short chain fatty acids.

Cotesia plutellae known as an endoparasitoid parasitizes larvae of the diamondback moth, Plutella xylostella which is a major pest in cruciferous crops. For the successful parasitization, maternal and embryonic factors of C. plutellae such as polydnavirus, ovarian proteins, teratocytes and venom are required. In this study, we identified calreticulin (Cp-CRT) gene from transcriptome data of the venom gland in C. plutellae. cDNA of CRT was cloned from total RNA of the venom gland via PCR and encodes 403 amino acids harboring several structural motifs such as a signal peptide sequence, a repetitive sequence, a putative coiled-coil sequence encompassing, and endoplasmic reticulum-recognizing domain (-KDEL). Phylogenetic analysis showed that the Cp-CRT gene formed a unique cluster with other hymenopteran CRT genes, indicating that the Cp-CRT belongs to the CRT family. To examine the physiological function of Cp-CRT, recombinant Cp-CRT, fused with 6X-His at N-terminal was constructed and expressed in E. coli. Recombinant Cp-CRT was successfully expressed via Western blot analysis and suppressed significant nodule formation when co-injected with E. coli as immune response inducer. These results suggest that the Cp-CRT involves in suppression of cellular immune response in the host

Matrix 2 protein ectodomain (M2e) of influenza virus appears to be a promising vaccine candidate because its sequence is highly conserved among virus strains. However, M2e is too meager to induce a strong immune response by itself. Several approaches are being used to increase the antigenicity of M2e. In an effort to enhance the M2e-specific immune response, we generated a TAT-conjugated M2e recombinant protein. Seven-week-old BALB/c mice were divided into three groups and transcutaneously immunized with 100 μg TAT-8×M2e (TAT conjugated with eight copies of M2e) and 8×M2e (eight copies of M2e) proteins on days 1, 15 and 29. The control mice were injected with PBS on the same days. Antibody titers specific for M2e were measured using indirect ELISA. Mice immunized with the TAT-8×M2e and 8×M2e proteins developed almost the same levels of M2e-specific total IgG and IgG1 antibodies. However, a higher level of M2e-specific IgG2a was observed in mice immunized with TAT-8×M2e than in those immunized with 8×M2e. These results suggest that TAT has an adjuvant effect that induces a Th1-type immune response. Therefore, the TAT-M2e vaccine can be applied to animals as a new influenza vaccine for enhancement of Th1-type immune responses.

Brucellosis is an important bacterial zoonosis in humans and domestic animals. Brucella spp. are taken up, and survive within non-professional and professional phagocytes. In common belief, diabetes mellitus increases susceptibility to pathogenic infection. In this study, Brucella (B.) abortus was inoculated into a diabetic animal model, db/db mice, in order to show the course of brucellosis in diabetic state. The liver proliferation, bacterial burden of the liver, level of cytokines in serum and macrophage migration into liver, were investigated at 14 days post-infection. In comparison with the uninfected control mice, the results revealed that the weight of the liver of infected db/db mice was higher but with lower bacterial load in this organ. The level of MCP-1 mRNA expression in the liver was lower, the levels of IL-12p70, IL-10, TNF-α and IFN-γ in serum was significantly higher and the macrophages migration was significantly lower in infected mice than in the control group. In conclusion, this present study suggested that MCP-1 suppression by B. abortus infection may inhibit the macrophages migration, and consequently may induce to abrogate the bacterial survival in db/db mouse liver. Furthermore, the increased inflammatory cytokines may contribute to inhibition of B. abortus proliferation in diabetic mice.

Insect blood cells, hemocytes, inhibit spreading behavior upon bacterial challenge to perform cellular immune responses. Hemocyte spreading is accomplished by cytoskeleton rearrangement, which is activated by various immune mediators, such as biogenic monoamins, plasmatocyte-spreading peptide(psp), and eicosanoids. However, little is known how these, immune mediators. acitvate hemocyte spreading behavior. A small G protein, Rac1, gene was identified in hemocytes of Spodoptera exiqua. Its expressed in most developmental stages accept egg and expecially expresses in hemocytes and fat body of Larval stage. In response to bacterial challenge, its expression was segnificantly up-regulated. However, RNA inteference (RNAi) of Rac1 expression inhibited hemocyte spreading behavior. under RNAi condition of Rac1, octopamine and psp failed to activate hemocyte spreading behavior. Interestingly, as addition of prostaglandinE2 to the RNAiconditioned Larval rescued the mediation of octopamine and psp. These results indicate that Rac1 is required for mediation of octopamine and psp on hemocytespreading behavior and suggest that Rac1 may activate eicosanoid biosnthesis.

As the immune reactions in human white blood cells of certain substances from insects to defend it when invaded by immune blood cells is increased. We experiment with changes in the total number of blood cells through the blood cells which increases and decreases, as well as to observe whether the immune response through any route is to evaluate what happens. Hemocyte population was analyzed in the last instar larvae of Spodoptera exigua. Granulocyte and plasmatocyte were predominant (>75%) types of hemocytes, whereas spherulocyte, prohemocyte, and oenocytoid hemocytes were observed in small densities (5~10%). Total hemocyte counts (THCs) were varied among different ages (day1-day5) of the last instar, in which day 3 larvae (L5D3) had the maximal density. Upon bacterial challenge to L5D3 larvae, THC was further enhanced within 2 h and then decreased to background level. This rapid THC increase in response to bacterial challenge was inhibited by injection with dexamethasone (1 ㎍ per larva). However, the addition of arachidonic acid reversed the inhibitory activity of dexamethasone and allowed the larvae to increase THC. This THC increase was mediated by cyclooxygenase products, but not by lipoxygenase products.

Phospholipase A2 (PLA2) is the committed catalytic step of eicosanoid biosynthesis, which has been a common molecular target of several entomopathogens to induce insect immunosuppression. Despite critical importance of PLA2 in insect immunity, its gene structure was not known. This study identified insect PLA2 gene associated with immune reactions in the red flour beetle, Tribolium castaneum. Based on a previous study that an immune-associated PLA2 in insect is secretory type of PLA2 (sPLA2), five highly matched cDNA sequences were obtained from T. castaneum genome database using an sPLA2 sequence probe encoded in Drosophila melanogaster. The expressions of these five putative PLA2 were confirmed by reverse transcriptase-polymerase chain reaction. Out of five genes, one PLA2 gene called TcPLA2B was chosen because it showed specific expression in hemocyte and fat body. TcPLA2B was cloned and expressed in Escherichia coli and its protein was purified. The purified TcPLA2B showed PLA2enzyme activity, which was specifically inhibited by bromophenacyl bromide (a specific sPLA2inhibitor) and dithiothreitol (reducing agent of disulfide bond). It was sensitive to pH (optimum at pH 6.0) and reaction temperature (optimum at 10-30°C), and calcium dependency. An immunofluorescence assay indicated that TcPLA2B was localized near to cellular membrane of the cytosol in the hemocytes of T. castaneum at immune chanlenge. Double-stranded RNA (dsRNA) of TcPLA2B-treated larvae showed knockdown of its mRNA expression and did not form hemocyte nodule formation, while control larvae could exhibit time- and bacterial dose-dependent nodule formation in response to bacterial challenge. Addition of arachidonic acid (the catalytic product of PLA2) to the dsRNA-treated larvae rescued the inhibition of nodule formation. These results suggest that TcPLA2B gene is associated with insect immune reaction.