Mycobacterium bovis Bacillus Calmette-Guerin (BCG) and BCG-based Vaccines Against Tuberculosis

차승빈,신성재 대한미생물학회 2014 Journal of Bacteriology and Virology Vol.44 No.3

Tuberculosis (TB) is the second leading infectious cause of mortality worldwide with about two million deaths peryear. The only licensed TB vaccine, Mycobacterium bovis bacillus Calmette-Guerin (BCG) shows limited protectionefficacy suggesting an improved vaccination strategy is required. Recently, several TB vaccine candidates have enteredclinical trials. These vaccine candidates are live mycobacterial vaccines designed to replace BCG or subunit vaccinesdesigned to boost immunity induced by BCG. Vaccines with different strategy such as therapeutic vaccines, which canalso be used in combination with drug therapy, are in the early stages of development to resolve latent TB or reactivationfrom the latent state. In this review, we discuss about development of BCG and BCG-based vaccines and further studiesnecessary for novel TB vaccine development to sterilize tuberculosis.

Bacteriophage의 Salmonella enterica serovar Enteritidis에 대한 in vitro 및 in vivo 효능 평가

차승빈,이원정,신민경,노유미,정명환,명길선,안영태,허철성,유한상,Cha, Seung-Bin,Rayamajhi, Nabin,Lee, Won-Jung,Shin, Min-Kyoung,Roh, Yu-Mi,Jung, Myung-Hwan,Myoung, Kil-Sun,Ahn, Young-Tae,Huh, Chul-Sung,Yoo, Han Sang 대한수의학회 2010 大韓獸醫學會誌 Vol.50 No.3

Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.

Brucella abortus 국내 분리주의 세포외막 단백질 분석 및 혈청학적 비교

차승빈,강미란,이원정,신민경,조동희,정석찬,유한상,Cha, Seung-Bin,Kang, Mi-Lan,Lee, Won-Jung,Shin, Min-Kyoung,Cho, Dong-Hee,Jung, Suk-Chan,Yoo, Han-Sang 대한수의학회 2008 大韓獸醫學會誌 Vol.48 No.4

Brucellosis is one of the most important zoonosis in worldwide. As one of the control measures, attempts have been made to develop new diagnostic methods using filed isolates as a national policy in many countries. Currently, bovine brucellosis in Korea have been received attention in both public health and economical aspects due to sudden increase of outbreak. Based on the situation, we compared standard strain (B. abortus 1119-3) with field isolates to reveal the differences among them. Biological and biochemical charateristics, antibiotic resistance profiles, outer membrane proteins (OMPs) and lipopolysaccharide analysis of the strains were included in this study. For the diagnostic purpose, an attempt was made to find out a novel antigen from the Korean isolates by serological analysis. There were differences about 55 kDa, 36-38 kDa and 20 kDa in analysis of OMPs by SDS-PAGE and Western blot with positive sera ($\geq$ 1:400 in SAT titer). Also, a serological diagnostic method, ELISA was conducted using OMPs of the strains as novel antigen. Relationships between O.D. and SAT titer were analyzed using field sera showing different SAT titer. High correlation coefficient was observed between SAT titer and ELISA. Results from this study suggested that a new diagnsotic method should be developed using their own field isolates in each country.

Predicting Genetic Traits and Epitope Analysis of apxIVA in Actinobacillus pleuropneumoniae

신민경,차승빈,Won-Jung Lee,유한상 한국미생물학회 2011 The journal of microbiology Vol.49 No.3

Actinobacillus pleuropneumoniae causes a severe hemorrhagic pneumonia in pigs. Fifteen serotypes of A. pleuropneumoniae express four different Apx toxins that belong to the pore-forming repeats-in-toxin (RTX)group of toxins. ApxIV, which is conserved and up-regulated in vivo, could be an excellent candidate for the development of a protective cross-serotype immunity vaccine, and could aid in the differential diagnosis of diseases caused by A. pleuropneumoniae. We identified and sequenced apxIVA from A. pleuropneumoniae serotype 2 isolated in Korea (Kor-ApxIVA). The Kor-ApxIVA was closely related to Switzerland (AF021919),China (CP000687), and China (GQ332268), showing 98.6%, 98.4%, and 97.2% amino acid homology,respectively. The level of amino acid homology, however, was higher than the nucleotide homology. The structural characteristics of ApxIVA showed RTX proteins, including N-terminal hydrophobic domains,signature sequences for potential acylation sites, and repeated glycine-rich nonapeptides in the C-terminal region of the protein. Thirty glycine-rich nonapeptides with the consensus sequence, L/V-X-G-G-X-G-N/D-D-X,were found in the C-terminus of the Kor-ApxIVA. In addition, the Kor-ApxIVA was predicted for the linear B-cell epitopes and conserved domains with determined peptide sequences. This genetic analysis of the Kor-ApxIVA might be an important foundation for future biological and functional research on ApxIVA.

Salmonella spp. 특이적인 검출을 위한 SYBR Green real-time PCR 기법 적용

신승원,차승빈,이원정,신민경,정명환,유안나,정병열,유한상,Shin, Seung Won,Cha, Seung Bin,Lee, Won-Jung,Shin, Min-Kyoung,Jung, Myunghwan,Yoo, Anna,Jung, Byeng Yeal,Yoo, Han Sang 대한수의학회 2013 大韓獸醫學會誌 Vol.53 No.1

The aim of this study was to applicate and evaluate a SYBR Green real-time PCR for the specific detection of Salmonella spp. Specificity of the PCR method was confirmed with 48 Salmonella spp. and 5 non-Salmonella strains using invA gene primer. The average threshold cycle ($C_T$) of Salmonella spp. was $11.83{\pm}0.78$ while non-Salmonella spp. was $30.86{\pm}1.19$. Correlation coefficients of standard curves constructed using $C_T$ versus copy number of Salmonella Enteritidis ATCC 13076 showed good linearity ($R^2=0.993$; slope = 3.563). Minimum level of detection with the method was > $10^2$ colony forming units (CFU)/mL. These results suggested that the SYBR Green real-time PCR might be applicable for the specific detection of Salmonella spp. isolates.

게르마늄 흑운모의 시험관 내에서의 E. coli와 Salmonella spp.에 대한 흡착력 및 성장 억제 효능 평가

정명환,차승빈,신승원,이원정,신민경,유안나,유한상,Jung, Myunghwan,Cha, Seung Bin,Shin, Seung Won,Lee, Won-Jung,Shin, Min-Kyoung,Yoo, Anna,Yoo, Han Sang 대한수의학회 2012 大韓獸醫學會誌 Vol.52 No.1

Germanium biotite, a natural mineral, has been used as a feed supplement to reinforce innate immune ability. The aim of the present study was to evaluate the effects of germanium biotite on the adsorptive and inhibition of growth abilities against Escherichia (E.) coli and Salmonella spp. in vitro. Two strains of enterotoxigenic E. coli and four strains of two Salmonella serotypes (Salmonella Derby and Salmonella Typhimurium), major bacterial diarrheal pathogens, were used for this experiment. The absorptive ability of germanium biotite against most Salmonella used in present experiment was observed weakly. The germanium biotite, however, showed significant effect of bacterial growth inhibition in most experiment bacteria. These results suggest that the use of the germanium biotite as feed supplement could alleviate diarrhea following inhibition of bacteria growth. It is also presumed that antibiotics usage for farm animals, considered as causes of antibiotic residue in meat and emerging antibiotic resistance, could be reduced through the use of germanium biotite as a feed supplement, in place of antibiotics used for the prevention of diarrhea.

Enterotoxigenic Escherichia coli 특이 박테리오파지 (ΦCJ19)의 사료 첨가제로서의 사양 평가

유안나,차승빈,신민경,박홍태,서효실,김재원,유한상,Yoo, Anna,Cha, Seung Bin,Shin, Min Kyoung,Park, Hong Tae,Seo, Hyo Sil,Kim, Jae Won,Yoo, Han Sang 대한수의학회 2013 大韓獸醫學會誌 Vol.53 No.2

Field efficacy of enterotoxigenic Escherichia coli-specific phage (${\Phi}CJ19$) as a feed additive was evaluated in weaning piglets. Fifty-four piglets at 3-4 weeks old were allocated in three different groups and two of them were fed with bacteriophage at different concentrations ($10^6PFU/kg$ feed and $10^8PFU/kg$ feed, respectively) for 30 days. Body weight and feed intake were measured at 10 days interval and body condition and fecal score were inspected every day. Based on the measurement, feed conversion rate (FCR) and average daily gain (ADG) of each group during 30 days were analyzed. The analysis suggests that the bacteriophage may help the improvement of FCR and ADG at $10^8PFU/kg$ of bacteriophage feeding group in 30 days. A result from analysis of fecal score indicates that the bacteriophage also may help to relieve the intermittent diarrhea in post-weaning stage. Those results suggest that bacteriophage might help the growth of piglets in post-weaning stage.

대두 생산과 활용 방안에 대한 연구

김광현,차승빈 한국지식정보기술학회 2023 한국지식정보기술학회 논문지 Vol.18 No.5

In this paper introduce about the soy bean and world output of soy been where and how to the soy bean be produced in producing the soy bean countries. The most produce soy bean country which is the Brazil and the most import soy bean country is the China. In the Korea soy bean is also very important for making the tofu, soy source and soy bean paste etc. Korea land is too small to cultivate soy bean in quantities, so Korea depend on importing the soy bean from cultivating soy bean countries. To slove this problem that rely on importing soy bean, Korea prepare self-sufficient soy bean in land policy. This policy we can know at last paragraph. Soy bean is the one of the important to the future practical use in Bio-energy of raw material and safe the earth environment problem by reducing the produce of meat from raise the livestock. Instead produce meat developing the Plant-based meat substitute is the more Eco-friendly to the environment. We can see how to soy bean is effected from economical cause and environmental cause. The Korea has the problem about the domestic supply soy bean because of the be small in land but Korea can solve this matter to prepare a supply measure that establish the building for only cultivate the soy bean like a grain. This is the best for the small country to cultivate the grains.

Development of a novel enzyme-linked immunosorbent assay to detect anti-IgG against swine hepatitis E virus

이원정,신민경,차승빈,유한상 대한수의학회 2013 JOURNAL OF VETERINARY SCIENCE Vol.14 No.4

Swine hepatitis E virus (HEV) is widespread throughout pigs in both developing and industrialized countries. This virus is an important zoonotic agent and a public concern worldwide. Infected pigs are asymptomatic, so diagnosing swine HEV relies on detection of the virus or antibodies against the virus. However, several obstacles need to be overcome for effective and practical serological diagnosis. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) that used a purified recombinant capsid protein of swine HEV. The potential clinical use of this assay was evaluated by comparing it with a commercial kit (Genelabs Technologies, Diagnostics, Singapore). Results of the ELISA were highly correlated with those of the commercial kit with a sensitivity of 97% and specificity of 95%. ROC (receiving operator characteristic) analysis of the ELISA data produced a value of 0.987 (95% CI, 0.977∼0.998, p < 0.01). The cut-off value for the ELISA was also determined using negative pig sera. In summary, the HEV-specific ELISA developed in the present study appears to be both practical and economical.

국내 우군에서 소 결핵 진단을 위한 피내검사법과 Interferon-γ 생성 검사의 비교

신승원,신민경,차승빈,우종태,이성모,구복경,조윤상,정석찬,유한상,Shin, Seung Won,Shin, Min Kyoung,Cha, Seung Bin,Woo, Jong Tae,Lee, Sung Mo,Ku, Bok Kyung,Cho, Yun Sang,Jung, Suk Chan,Yoo, Han Sang 대한수의학회 2011 大韓獸醫學會誌 Vol.51 No.2

Bovine tuberculosis (bTB), caused primarily by Mycobacterium bovis, continues to exert an economic loss, even in countries with active control measures, and is one of zoonotic diseases enable to be transmitted to human. The control and eradication of bTB are mainly based on a test and slaughter policy and/or abattoir surveillance. Various factors including limitation of diagnostic tests have been considered as major constraints to eradication. Single intradermal test (SIT) is the official diagnostic test. New diagnostic methods are needed to be developed, because of limitations of the test. In the present study SIT was compared with single intradermal comparative cervical test (SICCT) and interferon (IFN)-${\gamma}$ assay. There was very low correlation between SIT and SICCT. However, high correlation was shown between SIT and IFN-${\gamma}$ assay while no correlation was observed between SICCT and IFN-${\gamma}$ assay. Therefore, our results suggest the possibility of replacement of SIT with IFN-${\gamma}$ assay for the diagnosis of bovine tuberculosis.