Human hepatitis B virus (HBV) infection remains a serious public health problem worldwide, as it is one of the main risk factors for hepatocellular carcinoma (HCC). Cross-species transmission of HBV has been reported in non-human primates, and pigs may also be infected with HBV or an HBV-like agent. The purpose of this study was to demonstrate the presence of HBV antigens and anti-HBV antibodies in pig sera, providing further support for the existence of HBV or an HBV-like agent in pig populations. The HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) in pig serum samples were detected using HBsAg and HBeAg ELISA Kits, respectively. Antibodies to HBsAg and the Hepatitis B core antigen (HBcAg) in serum samples were also detected using anti-HBsAg and anti-HBcAg antibody ELISA Kits, respectively. HBsAg and HBeAg were detected in 7 of 442 (1.6%) and 7 of 184 (3.8%) pig serum samples, respectively. Furthermore, antibodies specific to HBsAg and HBcAg were identified in 45 of 442 (10.2%) and 39 of 434 (9.0%) pig serum samples, respectively. However, neither HBV DNA nor antibodies to HBeAg were detected in 409 and 298 pig serum samples, respectively. HBV antigens and anti-HBV antibodies were both present in a considerable number of pig serum samples, suggesting that pigs could be infected with a variant HBV or an HBV-like agent. Further studies will be necessary to confirm cross-species infection of pigs with HBV.

Mycoplasma hyorhinis (M. hyorhinis) is considered an etiological agent of arthritis in suckling pigs. Recently, some M. hyorhinis strains were shown to produce pneumonia that is indistinguishable from the mycoplasmosis caused by M. hyopneumoniae. In this study, we developed a sensitive and specific PCR assay to detect M. hyorhinis and applied the developed PCR assay for detection of Mycoplasma infection in clinical piglets infected with M. hyorhinis. We developed a new PCR assay using a M. hyorhinis-specific primer pair, Mrhin-F and Mrhin-R, designed from the Mycoplasma 16S-23S rRNA internal transcribed spacer (ITS) region. The primers and probe for the assay were designed from regions in the Mycoplasma 16S-23S rRNA ITS unique to M. hyorhinis. The developed PCR assay was very specific and sensitive for the detection of M. hyorhinis. The assay could detect the equivalent of 1 pg of target template DNA, which indicates that the assay was very sensitive. In addition, M. hyorhinis PCR assay detected only M. hyorhinis and not any other Mycoplasma or bacterial spp. of other genera. The new developed PCR assay effectively detected M. hyorhinis infection in pigs. We suggest that this PCR assay using a M. hyorhinis-specific primer pair, Mrhin-F and Mrhin-R, could be useful and effective for monitoring M. hyorhinis infection in pigs.

62세 여자가 약 7-10일간의 발열과 심와부 복통 및 오한으로 내원하여 단순한 급성 담석성 담낭염으로 진단받고 적극적인 시술과 약물치료를 받았으나, 임상경과가 호전되지 않고 비장경색의 합병증이 동반되어 추가 시행한 검사에서 삼일열 말라리아로 확진 받았다. 이후 절제된 담낭의 염증 소견은 말라리아 관련 급성 담낭염 소견을 시사하였다. 따라서 단순 급성 담석성 담낭염이라 하더라도 임상양상이 통상적인 회복소견을 보이지 않을 경우 다른 전신 질환에 합병된 담낭염의 가능성을 적극 고려해 보아야 한다.

A female wild raccoon dog was referred with a history of generalized seizure. Mild leukocytosis was noted on laboratory tests. Gross lesions included nasal hemorrhage, hemothorax, and hemorrhage in the urinary bladder with hematuria. Microscopically, interstitial and purulent bacterial pneumonia was observed in the lungs. In the cerebellum, characteristic eosinophilic intracytoplasmic and intranuclear inclusion bodies were found in Purkinje cells, and severe demyelination was observed in the cerebellar white matter. Canine distemper virus (CDV) infection was suspected and confirmed after detection of CDV nucleoprotein RNA in the cerebrum and the lungs by nested reverse transcription polymerase chain reaction (RT-PCR) and reverse transcription loop-mediated isothermal amplification (RT-LAMP). Based on the histopathological and molecular diagnostic findings, it was concluded that the raccoon dog was infected with CDV.

The effective use of disinfection and sterilization is important in preventing healthcare associated infections. Especially, multi-drug resistant organism and spore-forming bacteria are serious pathogens. Therefore, control of a nosocomial infection is emerging as a big problem. It is efficiently necessary to eliminate the pathogen. In this study, the components of sterilant were optimized. The optimized sterilant was applied for efficiency of sterilization using G. stearothermophilus E. coli, S. aureus as a standard microorganism. Our results were shown as follows : The decomposition or loss of potency of sterilant in small containers did not appeared for one month at ambient temperature. All of hydrogen peroxide 0.5%, peracetic acid 0.01%, and IPA 60% was effective for sterilization of E. coli, S. aureus using dilution method. Two kinds of sterilants(hydrogen peroxide 2%+peracetic acid 0.1%+IPA 0.5%, hydrogen peroxide 4%+peracetic acid 0.06%+IPA 1%) were bacteriocidal(> 6 log reduction). Microorganisms(E. coli, S. aureus) which treated with these sterilants for 2min were not grown in LB broth at 7 days culture. The gas plasma system for sterilization was applied. The sterilant solution(hydrogen peroxide 20%+IPA 5%) was vaporized in the sterilization chamber. The vaporized sterilant was bacteriocidal and sporocidal(> 6 log reduction) using G. stearothermophilus E. coli, S. aureus as biological indicators

Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

The Wolbachia bacterium is one of the best studied reproductive manipulators found in variuos invertebrate taxa. Theoretically it infects two thirds of invertebrates. It is thought to have a huge impact on its hosts’ evolution. The ant is a very successful organism in terrestrial ecosystems especially with its charicteristic sociality. However, the influence of Wolbachia on the animal remains to be paid more attention. This study is to examine Wolbachia infection frequency in some ant species in Korea. The result shows that 9 out of 12 ant species harbor the bacterium. Furthermore, at least two species show polymorphism in their infection status at the colony level. This will be used as a foundation of further study on the interactions between the bacterium and ant species.

Bromoviridae과 Cucumovirus속에 속하는 대표 바이러스인 오이모자이크바이러스 (Cucumber mosaic virus: CMV)는 많은 경제적으로 중요한 원예작물 및 관상식물들에 심한 손실을 초래하는 바이러스이다. 다중염기서열 비교는 현재까지 서브그룹1에 속하는 모든 CMV 계통들에서 3'말단부의 보전적 염기서열들이 존재하고 있음을 보여주었다. 이런 관찰에 기초하여, 우리는 CMV RNA3와 상동성을 가지는 162 bp 상보적 DNA를 포함하며 CMV감염에 대하여 식물 유래 RNA간섭 현상을 유도할 수 있는 도치된 반복 구조를 가지는 머리핀RNA (pIR-CMVNCR)를 발현시킬 수 있는 벡터를 제작하였다. 아그로박테리움을 이용하여 IR-CMVNCR의 일시 발현은 CMV 감염을 저해하였으며, 아그로박테리움이 접종된 식물의 상엽에서는 CMV 병징이 발현되지 않았다. 또한 RT-PCR결과는 아그로박테리움이 접종된 식물의 접종엽 및 상엽에서 CMV 서브유전자 4를 포함하는 CMV RNA들이 모두 검출이 되지 않았다. CMV 유래 작은 저해 RNA들의 축적이 관찰되었으며, 이의 결과의 의미는 아그로박테리움에 의해 IRCMVNCR을 일시 발현시킨 야생담배 (Nicotiana benthamiana)의 접종엽에서 RNA 간섭 현상이 유도되어 CMV 감염을 억제시키는 것으로 판명되었다.

The aim of the present study was to evaluate the use of an immunochromatographic test (ICT) strip using recombinant MPB70 (rMPB70) protein as a complementary tool for the diagnosis of naturally occurring tuberculosis in cattle. The study was performed on 249 cattle from populations known to be free from Mycobacterium bovis (M. bovis) and 119 cattle with M. bovis infection, confirmed postmortem. Compared to reference standards (culture isolation and/or visible lesion), the sensitivity of ICT was 94.12% (95% CI: 89.89∼98.35%) while the specificity was 96.80% (95% CI; 94.62∼96.82%). The findings indicate that the ICT strip is efficient for diagnosing bovine tuberculosis in cattle from Korea.

The Riptortus (stinkbug) has a specialized symbiotic organ, M4 midgut, to harboring symbiont Burkholderia. M4 midgut is located in abdomen and surrounded with insect hemolymph. Recently our group demonstrated that symbiotic Burkholderia showed different physiology after adapting in M4 gut compare with in vitro cultured Burkholderia. And population of symbiotic Burkholderia in the M4 midgut is regulated by special organ. However, the molecular mechanism to prevent spreading and migrating symbiont bacteria to other host tissues from symbiotic organ is not clear. Therefore, we assumed that symbiont Burkholderia are susceptible to host humoral immunity after established infection in M4 midgut to prevent spreading and migrating into the other host tissues through Riptortus hemolymph. To prove this assuming, we tested the susceptibility and survival rate of symbiont Burkholderia in hemolymph of Riptortus in vitro and in vivo. We also examined the susceptibility of symbiont Burkholderia using purified antimicrobial peptides (AMP), pyrrhocoricin-like, thanatin-like and defensin-like AMPs. Finally, we tested inducing ability for AMPs by systemic infection of symbiotic Burkholderia. Gene expression of purified AMPs was not different after systemic infection of both symbiont and in vitro cultured Burkholderia. Surprisingly, in vitro cultured Burkholderia resisted on bacteria injected hemolymph and purified AMPs but symbiont Burkholderia were highly susceptible in bacteria injected hemolymph and purified AMP. These results suggest that symbiont Burkholderia can't survive in the hemolymph after escaping symbiotic organ. Moreover, humoral immunity of host Riptortus is important to prevent spreading and migrating symbiont Burkholderia into the other host tissue or organ from symbiotic organ.

The phylogenetic analysis the queen polymorphic Vollenhovia emeryi ant reveals derivation of the Wolbachia- free short-winged from the Wolbachia-infected long-winged. However, intriguingly, some Japanese short-winged colonies harbor Wolbachia. Wolbachia specific bacteriophage (WO) is also detected in more than half of the infected colonies with no clear distribution pattern across the host insect lineage. We hypothesized that 1) the infected Japanese short-winged is in the intermediate stage to complete loss of Wolbachia and 2) the phage invaded the host after the host insect diverged. To test the hypotheses, we studied the strain diversity using the multi-locus sequence typing (MLST) of five ant colonies; three longwinged colonies from Korea and one long-winged colony and one short-winged colony from Japan. Both Korean and Japanese V. emeryi colonies show unexpectedly high level of Wolbachia strain diversity. However, the diversity is not significantly different between the long-winged and the short-winged against our first hypothesis. Phylogenies of Wolbachia show Korean strains and Japanese strains are largely monophyletic indicating prior infection before the host divergence. The strain diversity of the phage is also surprisingly high. Phylogenies of orf2 and orf7 genes are incongruent to that of Wolbachia and geographically distinct. This indicates that the phage is spatially static and the current infection pattern may be the consequence of local repeated gain and loss of the phage.

The ant species, Vollenhovia emeryi Wheeler (Hymenoptera: Myrmicinae) is endemic in East Asia and has invaded into North America. In this species, the queen caste shows polymorphism in its wing morphology; long-winged queen and short-winged queen, and two morphs are thought not to coexist in nature. This research is conducted to 1) deduce the phylogeographical structure of the two wing morphs in South Korea and to trace the distribution pattern from East Asia to North America, and 2) investigate the Wolbachia and WO phage infection frequency of the species. Either individuals or colonies of V. emeryi were collected from 80 locations, encompassing 68 locations in South Korea, 11 in Japan, and one in USA. Among the collected samples in South Korea, the long-winged morph is dominant and considered as the ancestral type, while the short-winged morph is very rare and derived character. The origin of the US population is neither Korea nor Japan at least in this study. However, we do not exclude the possibility that its origin is the other parts of Japan or the other countries. All of the long-winged morph are infected with Wolbachia, while the short-winged seems to be geographically partially infected. It suggests the possibility that the short wing trait is linked with the evolution of resistance to Wolbachia infection. Bacteriophage WO infection status has no correlation with host insect lineage.

Brucellosis is a zoonotic infectious disease of domestic animals, wild animals and humans. Innate immunity is a rapid and non-specific immune response that occurs during the early stages of Brucella invasion. Physical barriers such as epithelial cells and gastric juice secretions form the first line of defense. Humoral components such as complement and lysozyme can remove microorganisms by opsonization and bactericidal actions. Cellular components of the immune system, including macrophages, dendritic cells, neutrophils and innate T cells, have major roles in innate immunity. They recognize invading Brucella spp. by various cell surface receptors and then kill both the invading microorganisms and infected cells owing to their phagocytic or cytotoxic activity. In addition, they present Brucella antigens or produce cytokines to trigger adaptive immunity. Activated adaptive immunity consists of T helper cells, cytotoxic T cells and antigen-specific antibody-producing B cells. These can eliminate Brucella spp. effectively via antigen-specific mechanisms and by immunological memory. T cells activate bactericidal functions in macrophages by producing cytokines such as IFN-γ and by exerting cytotoxic effects on the infected cells. B cells produce antigen-specific antibodies that neutralize or opsonize the antigen. Because Brucella spp. can survive in macrophages and other host cells, Th-1 cellular immunity that enhances the bactericidal effects of phagocytic cells and the cytotoxic effects of lymphocytes is more important than humoral immunity in Brucella infection.

본 실험은 박과류(오이, 참외, 멜론)를 공시작물로 하여 친환경 육묘시 친환경 제제와 시용조건에 따른 흰가루병의 방제효과를 구명하고 기존 등록된 농약과 대조하여 그 효과를 검정하기 위해 수행되었다. 친환경제제는 유효 미생물로서 Ampelomyces quisqualis 94013(AQ94013)과 Bacillus subtilis Y1336(BS Y1336), 식물추출물로서 님오일(Neem oil)과 대황(Rheum undulatum)추출물, 미네랄 제제는 황수화제와 석회보르도액(Lime Bordeaux mixture)을 단용 또는 혼용처리하여 병 발생시기(발생전, 초기, 중기)에 따라 처리하였다. 모든 처리구에서 시설내 일 평균온도 30oC 이상, 하루 중 최고기온이 40oC 이상의 조건이 일주일 이상 지속되는 기간 동안 유묘의 흰가루 병의 발병도는 감소하였고, 대조구인 무처리구에서의 발병도도 크게 감소되어 10% 미만으로 나타났다. 발생 전 처리시 방제효과는 모든 작물에서 황수화제가 가장 높았고, BS Y1336는 방제가가 20~40%로서 가장 낮게 나타났다. 오이의 경우 발생전 처리시 님오일과 황수화제 처리는 살균제보다 높은 방제효과를 보였으며, 멜론에서는 농약과 비슷한 방제가를 보였다. 대황추출물은 전 생육 기간 동안 미생물제제보다 높은 방제가를 나타내었고, 오이와 멜론작물에서 발생초기 처리시 효과가 우수하였다. 물살포구는 무처리구와 비슷한 발병도를 보여 방제 효과가 없었고, 참외와 멜론에서 물살포 처리는 무처리보다 더 높은 발병도를 보였다. 병발생후 친환경제제 처리시 방제효과는 처리하는 시기가 늦어질수록 감소하였다. 친환경제제의 혼용처리에 의한 방제효과를 조사하였을 때, 대황추출물(1주차) + 황수화제(2주차) + 님오일(3 주차)의 조합으로 한 가지의 친환경제제를 주당 1회씩 총 3주 동안 혼용처리 했을 때, 전 육묘기간 동안 오이에서는 90% 이상, 모든 작물에서 평균 80% 이상의 방제가를 나타내어 다른 조합의 처리에 비해 높은 방제효과를 보였다. 본 실험결과를 통해 박과채소의 친환경 육묘시 흰가루병에 대한 친환경제제의 방제효과는 단용처리 했을 때, 병발생 전후 처리에 상관없이 모든 작물에서 황수화제 처리가 가장 높게 나타났으며, 혼용처리시에는 대황추출물(1주차) + 황수화제(2주차) + 님오일(3주 차)의 조합이 가장 효과가 높은 것으로 나타났다.

The present study was performed reviewing references determine the infection status of Korean freshwater fishes and relation with climate change of Korean peninsula. The most common infections of Korean freshwater fishes are flatworms (trematodes) and spiny-headed worms (acanthocephalans). Total 61 species of Korean freshwater fishes for infectious disease has been reviewed. Among them, 44 species of freshwater fish belong to Cyprinidae and 17 species for Cobitidae, Siluridae, Bagridae, Amblycipitidae, Synbranchidae, Centropomidae, Odontobutidae, Belontiidae, and Channidae. The Korean freshwater fishes had total 35 kinds of parasite pathogenic organisms (Acanthocephala opsariichthydis, Centrocestus sp., Cyathocotyle sp., Diplostomum orientale, Diplostomum sp., Echinochasmus sp., Echinostoma sp., Genarchopsis sp., Holostephanus metorchis, Metacercaria hasegawai, Metagonimus sp., Metorchis taiwanensis, Neoplagioporus zacconis, Palliolisentis chinanensis, Palliolisentis chunjuensis, Prosorhpynchus sp., Centrocestus armatus, Clonorchis sinensis, Holostephanus nipponicus, Metorchis orientalis, Cyathocotyle orientalis, Pseudexorchis major, Exorchis oviformis, Trachelobdella sinensis, Metagonimus miyata, Isoparorchis hypselobagri, Clinostomum complanatum, Centrocestus asadai, Metagonimus yokogawai, Echinostoma cinetorchis, Echinochasmus japonicus, Diplozoon nipponicum, Metagonimus takahashii, Carassotrema koreanum, and Echinostoma hortense). Maximum infections belonged to Pseudorasbora parva as Cyprinidae had 19 kinds of parasite organisms, including Centrocestus armatus, Clinostomum complanatum, Clinostomum complanatum, Clonorchis sinensis, Cyathocotyle orientalis, Echinochasmus japonicus, Exorchis oviformis, Holostephanus metorchis, Holostephanus nipponicus, Metacercaria hasegawai, Metagonimus yokogawai, Metorchis orientalis, Metorchis taiwanensis, Pseudexorchis major, Cyathocotyle sp., Diplostomum sp., Echinochasmus sp., Metagonimus sp. and Prosorhpynchus sp. Human foodborne trematode infections are an important public health concern in Korea. In the future, under the climatic extremes on Korean peninsula such as floods, drought and temperature variability, in particular, the susceptibility of freshwater fishes to infection will be decreased.

Peptidoglycan recognition proteins (PGRPs) are family of innate immune molecules that recognize bacterial peptidoglycan. PGRP-LE, a member of the PGRP family, selectively binds to diaminopimelic acid (DAP)-type peptidoglycan to activate both the immune deficiency (IMD) and proPhenoloxidase (proPO) pathways in insects. A PGRP-LE-dependent induction of autophagy to control Listeria monocytogenes has also been reported. We identified and partially characterized a novel PGRP-LE homologue, from Tenebrio molitor and analyzed its functional role in the survival of the insect against infection by a DAP-type PGN containing intracellular pathogen, L. monocytogenes. The cDNA is comprised of an open reading frame (ORF) of 990 bp and encodes a polypeptide of 329 residues. TmPGRP-LE contains one PGRP domain, but lacks critical residues for amidase activity. Quantitative RT-PCR analysis showed a broad constitutive expression of the transcript at various stages of development spanning from larva to adult. RNAi mediated knockdown of the transcripts followed by a challenge with L. monocytogenes showed a significant reduction in survival rate of the larvae, suggesting a putative role of TmPGRP-LE in sensing and control of L. monocytogenes infections in T. molitor. These results implicate PGRP-LE as a defense protein necessary for survival of T. molitor against infection by L. monocytogenes.

The intracellular endosymbiont bacterium Wolbachia is currently considered the most abundant in arthropods. Many Wolbachia manipulate host reproductive systems, it lead to male-killing, cytoplasmic incompatibility, parthenogenesis and feminization of genetic males with large impact on host ecology and evolution in arthropods. In this study, we investigated the distribution of Wolbachia infection in Ae. albopictus according to geographical distribution. We selected 9 areas and collected Ae. albopictus, these areas were distinguished with mountain chain and waterway. Whole genomic DNA were extracted from collected specimens and PCR analysis were accomplished in each specimen using Wolbachia specific primers such as 16S and WSP genes. As a results, we collected 730 Ae. albopictus at least 30 specimens in each area. Wolbachia infection rate showed different patterns between geographical region. Cheonbook, Cheonnam and Yeongdong showed 100% infection rate followed by Jeju (97%), Chungchung (88%), and Gyungnam (85%) with two Wolbachia marker genes. Our results indicated that Ae. albopictus are commonly infected with Wolbachia, it is possible that Wolbachia may act as endosymbiont in Ae. albopictus regardless of geographical region. Although, low infection rate of Wolbachia, we need continuous survey for the evaluation of Wolbachia strain within Ae. albopictus as a vector of dengue fever.

This study evaluated the efficacy and safety of medi- cal shampoo containing terbinafine hydrochloride and chlorhexidine gluconate in dogs with dermatophytos is complicated with bacterial infection. Although several stud- ies in dogs and cats with fungal dermatitis have found that oral administration of terbinafine is effective for controlling clinical signs, the topical form of terbinafine has rarely been studied in dogs and cats. Therefore, this study evaluated the efficacy of medical shampoo containing terbinafine hy- drochloride and chlorhexidine gluconate in dogs with der- matophytos is complicated with bacterial infection. Eight dogs infected with Microsporum spp. complicated with bacterial infection were enrolled in this study. These dogs were further blindly divided into Group 1 (no treatment, fourdogs) and group 2 (treated with medical shampoo with terbinafine/chlorhexidine, four dogs). Clinical improvement was evaluated using bacterial and fungal cultural evaluation combined with clinical evaluation. This study found that the medical shampoo has sufficient efficacy to treat varying degrees of dermatophytosis complicated with bacterial in- fection in dogs, although the speed of improvement differed according to the degree and type of infection. Our study also found that combined therapy using antifungal and antibac- terial agents can improve clinical signs more effectively and rapidly, suggesting that concurrent bacterial infection plays a significant role in the development of dermatitis.