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Mycobacterium avium-intracellulare complex (MAC) is one of the most prevalent pathogenic nontuberculous mycobacteria that cause chronic pulmonary disease. The prevalence of MAC infection has been rising globally in a wide range of hosts, including companion animals. MAC infection has been reported in dogs; however, little is known about interaction between MAC and dogs, especially in immune response. In this study, we investigated the host immune response driven by M. intracellulare using the co-culture system of canine T helper cells and autologous monocyte-derived macrophages (MDMs). Transcriptomic analysis revealed that canine MDMs differentiated into M1-like macrophages after M. intracellulare infection and the macrophages secreted molecules that induced Th1/Th17 cell polarization. Furthermore, canine lymphocytes co-cultured with M. intracellulare-infected macrophages induced the adaptive Th17 responses after 5 days. Taken together, our results indicate that M. intracellulare elicits a Th17 response through macrophage activation in this system. Those findings might help the understanding of the canine immune response to MAC infection and diminishing the potential zoonotic risk in One Health aspect.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease (JD), and it causes diarrhea and weakness in cattle. During a long subclinical stage, infected animals without clinical signs shed pathogens through feces. For this reason, the diagnosis of JD during the subclinical stage is very important. Circulating miRNAs are attracting attention as useful biomarkers in various veterinary diseases because of their expression changes depending on the state of the disease. Based on current knowledge, circulating miRNAs extracted from bovine serum were used to develop a diagnostic tool for JD. In this study, the animals were divided into 4 groups according to fecal shedding, the presence of antibodies, and clinical signs. Gene expression was analyzed by performing miRNA sequencing for each group, and it was identified that the miRNA expression changed more as the MAP infection progressed. The eight miRNAs that were differentially expressed in all infected groups were selected as biomarker candidates based on their significant differences compared to the control group. These biomarker candidates were validated by qRT-PCR. Considering the sequencing data, two upregulated miRNAs and two downregulated miRNAs showed the same trend in the validation results. Network analysis was also conducted and the results showed that mRNAs (IL-10, TGF-β1) associated with regulatory T cells were predicted to be activated in the subclinical stage. Taken together, our data suggest that two miRNAs (bta-miR-374b, bta-miR-2887) may play major roles in the immune response to MAP infection during the subclinical stage.

Paratuberculosis (PTB) is a chronic contagious granulomatous enteritis of wild and domestic ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). PTB causes considerable economic losses to the dairy industry through decreased milk production and premature culling. PTB-affected cattle undergo a subclinical stage without clinical signs and initiate fecal shedding of MAP into the environment. Current diagnostic tools have low sensitivity for the detection of subclinical PTB infection. Therefore, alternative diagnostic tools are required to improve the diagnostic sensitivity of subclinical PTB infection. In this study, we performed ELISA for three previously identified host biomarkers (fetuin, alpha-1-acid glycoprotein, and apolipoprotein) and analyzed their diagnostic performance with conventional PTB diagnostic methods. We observed that serum fetuin levels were significantly lowered in the subclinical shedder and clinical shedder groups than in the healthy control group, indicating its potential utility as a diagnostic biomarker for bovine PTB. Also, fetuin showed an excellent discriminatory power with an AUC = 0.949, a sensitivity of 92.6%, and a specificity of 94.4% for the detection of subclinical MAP infection. In conclusion, our results demonstrated that fetuin could be used as a diagnostic biomarker for enhancing the diagnostic sensitivity for the detection of subclinical MAP infections that are difficult to detect based on current diagnostic methods.

Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne’s disease, which is a chronic granulomatous enteropathy in ruminants. Determining the genetic diversity of MAP is necessary to understand the epidemiology and biology of MAP, as well as establishing disease control strategies. In the present study, whole genome-based alignment and comparative analysis were performed using 40 publicly available MAP genomes, including newly sequenced Korean isolates. First, whole genome-based alignment was employed to identify new genomic structures in MAP genomes. Second, the genomic diversity of the MAP population was described by pangenome analysis. A phylogenetic tree based on the core genome and pangenome showed that the MAP was differentiated into two major types (C- and S-type), which was in keeping with the findings of previous studies. However, B-type strains were discriminated from C-type strains. Finally, functional analysis of the pangenome was performed using three virulence factor databases (i.e., PATRIC, VFDB, and Victors) to predict the phenotypic diversity of MAP in terms of pathogenicity. Based on the results of the pangenome analysis, we developed a real-time PCR technique to distinguish among S-, B- and C-type strains. In conclusion, the results of our study suggest that the phenotypic differences between MAP strains can be explained by their genetic polymorphisms. These results may help to elucidate the diversity of MAP, extending from genomic features to phenotypic traits.

Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne’s disease, which is a chronic and debilitating disease in ruminants. MAP is also considered to be a possible cause of Crohn’s disease in humans. However, few studies have focused on the interactions between MAP and human macrophages to elucidate the pathogenesis of Crohn’s disease. We sought to determine the initial responses of human THP-1 cells against MAP infection using single-cell RNA-seq analysis. Clustering analysis showed that THP-1 cells were divided into seven different clusters in response to phorbol-12-myristate-13-acetate (PMA) treatment. The characteristics of each cluster were investigated by identifying cluster-specific marker genes. From the results, we found that classically differentiated cells express CD14, CD36, and TLR2, and that this cell type showed the most active responses against MAP infection. The responses included the expression of proinflammatory cytokines and chemokines such as CCL4, CCL3, IL1B, IL8, and CCL20. In addition, the Mreg cell type, a novel cell type differentiated from THP-1 cells, was discovered. Thus, it is suggested that different cell types arise even when the same cell line is treated under the same conditions. Overall, analyzing gene expression patterns via scRNA-seq classification allows a more detailed observation of the response to infection by each cell type.

Johne’s disease (JD) caused by Mycobacterium avium subsp. paratuberculosis (MAP) is a chronic, wasting infectious disease in ruminants that causes enormous economic losses to the dairy and beef cattle industries. Understanding the mechanism of persistency of MAP is key to produce novel ideas for the development of new diagnostic methods or prevention techniques. We sought interactions between the host and MAP using epithelial passage model, which mimic initial stage of infection. From the transcriptomic analysis of bovine immune cells (PBMCs), it was suggested that infection through the epithelial cells elicited prolonged Th17-derived immune response, as indicated by upregulation of IL-17A, IL-17F and RORC until 120 h p.i., compared to directly infected PBMCs. Global downregulation of gene expression was observed after 72 h p.i., especially for genes encoding cell surface receptors of phagocytic cells, such as Toll-like receptors and MHC class II molecules. In addition, the cholesterol efflux transporters ABCA1, ABCG1, and APOE, which are regulated by the LXR/RXR pathway, were downregulated. In summary, it would be suggested that the host initiate immune response to activate Th17-derived cytokines, and MAP survives persistently by altering the host adaptive immune response by suppressing surface receptors and manipulating lipid metabolism in phagocytic cells.

Non-tuberculous mycobacteria (NTM) are ubiquitous microorganisms that have the potential to cause disease in both humans and animals. Recently, NTM infections have rapidly increased in South Korea, especially in urbanized areas. However, the distribution of species and the antibiotic resistance profile of NTM in environmental sources have not yet been investigated. Therefore, we analyzed the distribution of species and the antibiotic resistance profile of NTM in soil within urban areas of South Korea. A total of 132 isolates of NTM were isolated from soil samples from 1 municipal animal shelter and 4 urban area parks. Among the 132 isolates, 105 isolates were identified as slowly growing mycobacteria (SGM) and 27 isolates as rapidly growing mycobacteria (RGM) based on the sequences of the rpoB and hsp65 genes. The antibiotic resistance patterns of NTM isolates differed from species to species. Additionally, a mutation in the rrs gene found in this study was not associated with aminoglycoside resistance. In conclusion, our results showed that NTM isolates from South Korean soil exhibit multidrug resistance to streptomycin, amikacin, azithromycin, ethambutol, isoniazid, and imipenem. These results suggest that NTM may pose a public threat.

The aim of this study was to describe the genetic diversity of Mycobacterium avium subsp. paratuberculosis (MAP) obtained from individual cows in Korea. Twelve MAP-positive fecal DNA samples and 19 MAP isolates were obtained from 10 cattle herds located in 5 provinces in Korea. In addition, 5 MAP isolates obtained from the Czech Republic and Slovakia and 3 isolates from Australia were genotyped for comparison with the domestic isolates. The most prevalent strains in Korea were of the "bison-type" genotype (23 of 31 fecal DNA/isolates) and were distributed nationwide. The remaining MAP isolates (8) and all of the foreign isolates were identified as "cattle-type". The bison-type strains which were discriminated only as INMV 68 in variable-number tandem repeats of mycobacterial interspersed repetitive units (MIRU-VNTR) typing. Multilocus short sequence repeat (MLSSR) typing differentiated the bison-type strains into 3 different subtypes. The cattle-type strains were divided into 3 subtypes by MIRU-VNTR and 8 subtypes by MLSSR. The allelic diversities in the MIRU-VNTR and MLSSR results were calculated as 0.567 and 0.866, respectively. These results suggest that MIRU-VNTR typing cannot provide a sufficient description of the epidemiological situation of MAP. Therefore, an alternative method, such as MLSSR, is needed for typing of MAP strains to elucidate the molecular epidemiology of MAP infections. Overall, this study is the first epidemiological survey report in Korea using both MIRU-VNTR and MLSSR typing methods, and it has provided basic data necessary to elucidate the characteristics of MAP infections in Korea.

Thermal conditions are an important environmental factor in maintaining healthy pigs because they affect feed intake, growth efficiency, reproduction and immune responses in pigs. RAVI, a regenerative far-infrared heating system, can effect pig production by emitting an optimal far-infrared wavelength. Far-infrared radiation has been reported to increase microvascular dilation and vascular flow volume. The purpose of this study was to evaluate the immunobiological differences between pigs raised with the RAVI system and the gasoline heater system. Twenty-six-week-old weaned pigs were raised in two rooms that were equipped with a RAVI system or a gasoline heater for 8 weeks. A porcine atrophic rhinitis vaccine was administered after two weeks and transcriptome analysis in whole blood were analyzed at 2-week intervals. Signaling pathway analyses of the RAVI group at 8 weeks showed the activation of pathways related to nitric oxide (NO) production. This suggests that the application of RAVI might induce the production of NO and iNOS, which are important for increasing the immune activity. Similar to the result of microarray, phenotypic changes were also observed at a later period of the experiment. The increase in body weight in the RAVI group was significantly higher than the gasoline heater group at 8 weeks. The antibody titer against the vaccine in the RAVI group was also higher than that the gasoline heater group at 4 weeks and 8 weeks. This evaluation of the use of a far-infrared heating system with pigs will be helpful for applications in the pig farm industry and pig welfare.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of bovine paratuberculosis (PTB). The first step in the control of PTB is the identification and isolation of sub-clinical fecal shedders from the herd. In the current study, real-time and nested PCR targeting MAP-specific genetic elements (IS900 and ISMAP02) DNA isolated from fecal samples were used to detect MAP infection in cattle. Of the 1,562 fecal samples obtained from 37 herds, regardless of diarrhea, 35 samples tested positive in both IS900-targeted real-time and ISMAP02-targeted nested PCR. At the herd level, 12 of the 37 herds were found to be positive for MAP. Detection rates of the PCR tests were similar to those reported for ELISA-based methods. These results suggest that PCR can be used to detect sub-clinical fecal shedders of MAP.

Infection with Brucella abortus causes contagious zoonosis, brucellosis, and leads to abortion in animals and chronic illness in humans. Chitosan nanoparticles (CNs), biocompatible and nontoxic polymers, acts as a mucosal adjuvant. In our previous study, B. abortus malate dehydrogenase (Mdh) was loaded in CNs, and it induced high production of pro-inflammatory cytokines in THP-1 cells and systemic IgA in BALB/C mice. In this study, the time-series gene expression analysis of nasal-associated lymphoid tissue (NALT) was performed to identify the mechanism by which Mdh affect the target site of nasal immunization. We showed that intranasal immunization of CNs-Mdh reduced cell viability of epithelial cells and muscle cells at first 1 h, then induced cellular movement of immune cells such as granulocytes, neutrophils and lymphocytes at 6h, and activated IL-6 signaling pathway at 12h within NALT. These activation of immune cells also promoted signaling pathway for high-mobility group box 1 protein (HMGB1), followed by the maturation of DCs required for mucosal immunity. The CNs also triggered the response to other organism and inflammatory response, showing it is immune-enhancing adjuvant. The ELISA showed that significant production of specific IgA was detected in the fecal excretions and genital secretions from the CNs-Mdh-immunized group after 2 weeks-post immunization. Collectively, these results suggest that B. abortus Mdh-loaded CNs triggers activation of HMGB1, IL-6 and DCs maturation signaling within NALT and induce production of systemic IgG and IgA.

The aim of this study was to investigate the induction of mucosal immune responses by an important Brucella abortus antigen, malate dehydrogenase (Mdh), loaded in mucoadhesive chitosan nanoparticles (CNs) and immunized intranasally in a BALB/c mouse model. The production of cytokines was investigated in human leukemic monocyte cells (THP-1 cells) after stimulation with the nanoparticles. Mdh-loaded CNs (CNs-Mdh) induced higher interleukin (IL)-6 production than unloaded antigens and TF loaded CNs (CNs-TF). Using ELISpot to quantify cytokines and antibody-secreting cells in the intranasally immunized mice, IL-4 and IgG-secreting cells were found to be significantly increased at 4 weeks and 6 weeks post-immunization in the CNs-Mdh immunized group, respectively. Increases in Mdh-specific IgG, IgG1, and IgG2a antibodies were confirmed at 6 weeks after immunization, indicating a predominant IgG1 response. Analysis of the mucosal immune response in the intranasally immunized mice revealed, Mdh-specific IgA and total IgA in the nasal washes, genital secretions, fecal extracts and sera that were remarkably increased in the CNs-Mdh-immunized group compared to the CNs-TF-immunized group except total IgA of nasal wash. Therefore, the results indicated that the intranasal immunization of CNs-loaded B. abortus Mdh antigen effectively induced antigen-specific mucosal immune responses through the elicitation of Th2-related immune responses.

Johne's disease is a chronic wasting disease of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP), resulting in inflammation of intestines and persistent diarrhea. The initial host response against MAP infections is mainly regulated by the Th1 response, which is characterized by the production of IFN-γ. With the progression of disease, MAP can survive in the host through the evasion of the host's immune response by manipulating the host immune response. However, the host response during subclinical phases has not been fully understood. Immune regulatory genes, including Th17-derived cytokines, interferon regulatory factors, and calcium signaling-associated genes, are hypothesized to play an important role during subclinical phases of Johne's disease. Therefore, the present study was conducted to analyze the expression profiles of immune regulatory genes during MAP infection in whole blood. Different expression patterns of genes were identified depending on the infection stages. Downregulation of IL-17A, IL-17F, IL-22, IL-26, HMGB1, and IRF4 and upregulation of PIP5K1C indicate suppression of the Th1 response due to MAP infection and loss of granuloma integrity. In addition, increased expression of IRF5 and IRF7 suggest activation of IFN-α/β signaling during subclinical stages, which induced indoleamine 2,3-dioxygenase mediated depletion of tryptophan metabolism. Increased expression of CORO1A indicate modulation of calcium signaling, which enhanced the survival of MAP. Taken together, distinct host gene expression induced by MAP infection indicates enhanced survival of MAP during subclinical stages.

Escherichia (E.) coli is commensal bacteria found in the intestine; however, some pathogenic strains cause diseases in animals and humans. Although E. coli does not typically produce hydrogen sulfide (H2S), H2S-producing strains of E. coli have been identified worldwide. The relationship between virulence and H2S production has not yet been determined. Therefore, characteristics of H2S-producing isolates obtained from swine feces were evaluated including antibiotic resistance patterns, virulence gene expression, and genetic relatedness. Rates of antibiotic resistance of the H2S- producing E. coli varied according to antibiotic. Only the EAST1 gene was detected as a virulence gene in five H2S-producing E. coli strains. Genes conferring H2S production were not transmissible although the sseA gene encoding 3-mercaptopyruvate sulfurtransferase was detected in all H2S-producing E. coli strains. Sequences of the sseA gene motif CGSVTA around Cys238 were also identical in all H2S- producing E. coli strains. Diverse genetic relatedness among the isolates was observed by pulsed-field gel electrophoresis analysis. These results suggested that H2S-producing E. coli strains were not derived from a specific clone and H2S production in E. coli is not associated with virulence genes.

(Received: April 18, 2015; Accepted: May 11, 2015)Abstract : Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is one of the mostwidespread and economically important diseases in cattle. Current diagnostic methods are based on the detection ofanti-MAP antibodies in serum or isolation of the causative agent. However, these techniques are often not applicablefor cases of subclinical infection due to relatively low sensitivity. Therefore, finding new antigen candidates that stronglyreact with the host immune system had been attempted. To effectively detect infection during the subclinical stage,several antigen candidates were selected based on previous researches. Characteristics of the selected antigen candidateswere analyzed using bioinformatics-based prediction tools. A total of nine antigens were selected (MAP0862,MAP3817c, MAP2077c, MAP0860c, MAP3954, MAP3155c, MAP1204, MAP1087, and MAP2963c) that have MAP-specific and/or high immune responses to infected animals. Using a transmembrane prediction tool, five of the nineantigen candidates were predicted to be membrane protein (MAP3817c, MAP3954, MAP3155c, MAP1087, andMAP1204). Some of the predicted protein structures identified using the I-TASSER server shared similarities withknown proteins found in the Protein Data Bank database (MAP0862, MAP1204, and MAP2077c). In future studies,the characteristics and diagnostic efficiency of the selected antigen candidates will be evaluated.Keywords : antigen, bioinformatics, discovery, Johne’s disease

Mycobacterium avium subsp. paratuberculosis (MAP) causes paratuberculosis or Johne"s disease, an intestinal granulomatous infection in domestic and wild animals. The study aimed to develop and evaluate a panel of multiplex quantitative real-time polymerase chain reaction (mqPCR) assay for simultaneous detection of three MAP-specific genes (IS900, F57 and ISMAP02 genes). The analytic sensitivity (i.e., limit of detection, expressed as cells per 1 ml) was 150 for IS900, 1500 for F57, and 50 for ISMAP02. The specificity of the method was determined by testing 152 bovine fecal samples. Based on the test, it showed that the assay simultaneously detected the target genes in short period of time and at lower cost compared to laboratory routine tests. The test agreement between the assay and routine test was 94%. The discrepancy in the results was due to samples that were tested positive by the panel but negative by the routine tests, suggesting that the assay has higher sensitivity than the routine tests. In conclusion, the mqPCR assay could be a rapid and accurate testing tool for investigating paratuberculosis or Johne"s disease cases in domestic and wild animals.

Mycobacterium avium subsp. paratuberculosis (MAP) is a causative agent of Johne's disease or paratuberculosis (PTB), which is a chronic debilitating disease in ruminants, that is characterized by incurable enteritis and persistent diarrhea. ISMap02 is one of the major targets of PCR because it is present in multicopies (six copies) and known to be specific to MAP. However, in the present study, non-MAP mycobacteria were shown to be positive by ISMap02 targeting PCR. Two bacterial isolates (Sample ID: BO-038 and BO-042) were cultured from bovine fecal samples that produced positive results in three of two ISMap02 targeting PCR analyses with negative results in IS900 real-time PCR. Species identification using 16S rRNA gene sequencing and hsp65 gene partial sequencing revealed that strains BO-038 and BO-042 were M. virginiense and M. nonchromogenicum, respectively, which both belong to the M. terrae complex (MTC). Moreover, the two isolates shared a novel insertion sequence (IS) with high similarity to some parts of nucleotide sequences of ISMap02, and IS was presumed to be identical to that present in M. heraklionense. Both the novel IS and ISMap02 were characterized as IS1182 family members, and several sequences similar to ISMap02 were identified by BLAST analysis. In addition, the DDE transposase of the novel IS showed great similarity in the N-terminal portion with the IS5/1182 DDE transposase of other mycobacteria. These results suggest that ISMap02 has a conserved region with similarity to other ISs, and that the diagnostic value of the primer sets targeting that region should be re-addressed.

A 1-year-old, female domestic shorthair cat was presented with anorexia, depression and weight loss, accompanied by multifocal nodules affecting the face, pinnae and periarticular tissue. Routine medical treatments were ineffective. The animal's physical condition continued to deteriorate and it finally died. Post-mortem examination revealed multifocal to coalescing firm nodules with occasional ulceration affecting the ears, peri-ocular areas, nasal planum, oral cavity and laryngopharyngeal region. Tan-coloured, firm, nodular lesions were also observed in the periarticular tissue, lungs and tracheobronchial and mediastinal lymph nodes. Impression smears of several of these lesions revealed a myriad of slender rod-shaped organisms, mainly in the cytoplasm of macrophages. Histopathological examination showed severe pyogranulomatous inflammation with or without necrosis in the nodules. Acid-fast staining revealed large numbers of acid-fast bacilli. Mycobacterium kansasii was detected in the tissues using multiplex polymerase chain reaction and DNA sequencing. No protozoal or fungal organisms were detected using special stains. On the basis of these results, the cat was diagnosed with systemic M. kansasii infection. To our knowledge, there have been few reports of M. kansasii infection, especially with systemic spread, in cats.

본 연구는 설계 과정의 분해, 시공 과정의 조립, 공사비 적산 등 공사의 계획과 관리에 걸친 모든 공사 관리의 업무를 체계화할 수 있는 공동주택의 공사정보분류체계를 제시 하였다. 또한, 본 연구는 이 공사정보분류체계를 작업순서에 따라 관계형 데이터베이스 (Data Base)로 구축 방법을 제시하였고, 구축된 데이터베이스를 근거로 적산 자동화 시스 템 개념 모형을 구축하였다. 이러한 적산 자동화 시스템 개념 모형은 기존 적산 시스템들 의 근본적인 문제점이었던 부적절함을 해소하여 공동주택 건설현장에서 효과적으로 적용 가능한 과학적인 적산 시스템으로 활용할 수 있을 것이다. 공동주택, 공사정보분류체계, 개념 모형, 적산 자동화, 관계형 데이터베이스 c 2014 Korea Society of Disaster Information All rights reserved 공동주택의 공사정보분류체계를 활용한 적산 자동화 개념 모형 개발 A Conceptual Model for Automated Cost Estimating Using Work Information Classification System of Apartment House Lee Yang Kyu, Park Hong Tae a Department of Civil Environmental Daelim University, 29 imgokro, Dongangu, anyang-si Gyeonggi-do, 431-715, Republic Korea b Department of Civil Engineering, Kongju National University, 275 budae-dong Seobuk-gu, Cheonan-si, Chungnam 331-717, Republic Korea 1 Tel. 82-31-467-4917. Fax. 82-31-467-4910 Email. yklee@daelim.ac.kr * Corresponding author Tel. 82-41-521-9310. Fax. 82-41-521-9310, Email:htpark@kongju.ac.kr ARTICLE HISTORY Recieved Jan. 27, 2014 Revised Feb. 12, 2014 Accepted Mar. 10, 2014 Journal of the Korea Society of Disaster Information Vol.10 No.1 ❙ pp. 15 24 Available online at www.kosdi.or.kr 1976-2208 c 2014 Korea Society of Disaster Information All rights reserved. Journal of the Korea Society of Disaster Information. Vol.10, No.1, pp.15 24

This study attempted to estimate construction costs in accordance with the resource-based cost estimation (unit cost price) system by diameter for TBM method, and analyzed the direct cost and the total cost. Based on such figures, this study performed a regression analysis and proposed a model for an equation for estimating construction costs. model for the resource-based cost estimation (unit cost price) system classified by diameter for TBM method proposed by this study can be effectively applied to business planning, preliminary investigation, feasibility study, construction cost estimations in the early design stages.

Inspection system which was performed in the past selected the final winning bidder by separating Pre-Qualification, the basic design evaluation and price evaluation. However, turnkey and alternative bidding construction has selected way to determine the successful bidder as the best design and the most low cost method, comprehensive evaluation method (bid price adjustment, design score adjustments, weights standard), the best design and the fixed amount method from january 1, 2010 to the present. Due to these institutional problems, orders institutions were often a way to determine the successful bidder to apply differently for identical or similar construction. Therefore this study winning bidder selection criteria through analysis of construction the number and construction order performance, order performance by work division and institutional orders, the weights bidder order performance and orders status of turnkey alternative tendering construction to civil part performed in domestic.

In this study, a work information classification system standard for TBM tunnel construction is presented, and based on this standard, a work information classification system is constructed hierarchically based on DB structure. The proposed system divides into facility classification level, functional component classification level, work classification level and resource classification level hierarchically.

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of bovine paratuberculosis (PTB). The first step in the control of PTB is the identification and isolation of sub-clinical fecal shedders from the herd. In the current study, real-time and nested PCR targeting MAP-specific genetic elements (IS900 and ISMAP02) DNA isolated from fecal samples were used to detect MAP infection in cattle. Of the 1,562 fecal samples obtained from 37 herds, regardless of diarrhea, 35 samples tested positive in both IS900-targeted real-time and ISMAP02-targeted nested PCR. At the herd level, 12 of the 37 herds were found to be positive for MAP. Detection rates of the PCR tests were similar to those reported for ELISA-based methods. These results suggest that PCR can be used to detect sub-clinical fecal shedders of MAP.

The aim of shotcrete is to increase the bearing capacity and to protect the excavated surface from erosion by preventing falling of rock mass. Shotcreting method is divided into two types as dry process and wet process. Since 1997, wet process method has been used more frequently than dry process method in field works. The failure to bond, so called rebound, occurs in many case during shotcrete works. The excess amount of rebound has a significant effect on the total construction cost. For example, material and craft-man cost increases, the shooting time delays due to deceleration of work execution stage, work efficiency of craft-man decreases and additional cost to remove the muck generates. In this study, therefore, the experimental analysis of rebound amount and strength was conducted by analyzing the actual construction data for wet process type of shotcreting method upon accelerator contents. Also, the effective and rational method was suggested, which can be actually implemented in the Korea construction sites.

This research collected work drilling report of TBM method site developed by WRITH company to TBM equipment company in Germany and analyzed work operating productivity. Based Oil the data analyzed TBM operating productivity, This research derived and presented excavation speed (m/day) by TBM diameter (3.0m, 3.5m, 3.8m) and rock. Also, based on the excavation speed (m/day) by TBM diameter, This research estimated a day direct construction cost and total direct construction costs by applying a direct construction cost which spent on per 1m. When we perform a similar geological construction in the future, excavation speed and direct construction cost which were derived by TBM diameter and rock is thinking the effective utilization data to estimate construction cost and plan schedule management before the start of construction.

The schedule management in construction project have been performed independently by two method, WBS base management and CBS based management, However, the integrated management model is needed according to efficiency of construction project management and introduction of new EVMS system of KOREA government. For this purpose, interworking database system is designed through efficient modeling of WBS based management data and CBS based management data and relation establishment between entities in this paper. WBS based management DB consists of 4 entities and CBS based management DB consists of 3 entity and one of the 4 WBS DB entities is used to play interworking role between WBS and CBS, Also, attributes of all entities are well defined so that data consistency, integrity and newest are guaranteed. The designed interworking database model was realized using Nex-Pert Pro and the efficiency of data interworking was proved.

Turnkey and alternative-design contracts was introduced operation in the country from january 2004 to February 2010. this was introduced operation. The selecting method of contractor for turnkey projects has being operated by an open forum type that consists of technical committee and appraisal committee in an integrated design approach deliberations. But to improve the operation of the problems, the government is operating design review committee consisting of the central, local and special design advisory committee in march 2010. But this system has also been suggested as still problems, too many contracts by large companies, increasing of design cost and excessive competition. This study is intended to contribute that turnkey and alternative-design contracts can be well operated in domestic construction markets. In this paper suggests some improvement methods and operational problems caused from application of an open forum type for selecting contractor of turnkey project.