본 연구는 다양한 항생제 내성을 갖는 Salmonella enterica subsp. enterica serovar Typhimurium ATCC 19585 (STWT), S. Typhimurium KCCM 40253 (STKCCM), ciprofloxacininduced S. Typhimurium ATCC 19585 strains (STCIP), and S. Typhimurium CCARM 8009 (STCCARM)에 대한 phage의 흡착 및 용균 특성을 평가하였다. PBST-10, PBST-13, PBST-32, PBST-35, P-22, P-22 B1 phages는 narrow host range를 보였다. 숙주인 STWT, STKCCM, STCIP에 대한 phage 의 흡착률은 각각 47-85%, 58-95%, 61-93%였지만, STCCARM 에 대한 phage의 흡착률은 14-36%의 낮은 수준을 보였다. STWT, STKCCM, STCIP, STCCARM에 대한 phage burst size는 각 각 43-350, 37-530, 66-500, 24-500 plaque-forming unit (PFU)으로 다양하게 관찰되었다. P-22 B1을 제외한 모든 phage는 배양 초기에 STCIP숙주를 효과적으로 저해하였다. 이러한 결과는 항생제 내성균을 저해하기 위해 phage control system 개발에 유용한 정보로 활용될 것이다.

Salmonella Enteritidis (SE) infection, one of a major foodborne disease remains considerable public health concerns globally. In this study, we have constructed that the genetically inactivated SE ghost vaccine candidate, JOL2220 (Δlon ΔcpxR Δasd), which express the immunogenic hemagglutinin portion (HA1) of H1N1 virus on the Salmonella surface. The lysis gene cassette, holin-endolysin system originated from double DNA bacteriophage λ was cloned into the ghost vector plasmid pJHL464, which activated under the control of the convergent promoter system. Scanning electronic microscopic examination revealed that transmembrane tunnels of JOL2220 was formed following the 48 hr of the lysis mediated by the holin-endolysin system. In vitro expression of HA1 antigenic protein was also determined by Western blot analysis. The Chickens immunized with JOL2220 ghost via intramuscular and oral routes showed significant increase in IgG level against both SE and HA1 antigen. Further, we observed markedly enhancement of lymphocyte proliferation and T-cell differentiation in the primed PBMC of the immunized chicken, compared to those in the control group. Collectively, the results indicated that the SE ghosts expressing HA1 antigen may have immunostimulatory properties against Salmonella and influenza infection where the chicken is the predominant reservoir.

Earthworms, especially Eisenia andrei, are cultivated for a variety of purposes including waste disposal and compost production. In this study, bacteria from earthworm gut were cultured and the dominant species identified. Subsequently, we isolated bacteriophages able to lyse the isolated gut bacteria. Two dominant genera of gut bacteria, Aeromonas sp. and Citrobacter sp. were identified by using MALDI-TOF MS analysis and a library was constructed to find lytic phages. Phage EF1 showed lytic activity for C. freundii and two Citrobacter isolates, C. braakii and C. murliniae. These 3 species have similar sensitivities to EF1. Several aspects of the life cycle of EF1 were investigated by using C. freundii under optimal growth conditions. EF1 infects C. freundii with a moderate latent period, approximately 25 min, and a large burst size averaging 5 × 109 per infected cell. Moreover, the antimicrobial activity of EF1 was well maintained under diverse conditions including a broad temperature range of 40°C to 50°C and a wide pH range of 4 to 11. In conclusion, the results indicate that earthworm casting contain a wide range of bacteria species, for which there are various corresponding bacteriophages.

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.

벼 흰잎마름병균의 정확한 진단을 위하여 PCR용 진단 kit를 개발하였다. 본 PCR kit를 개발하기 위하여 벼 흰잎마름 병균 유전체 정보 중 phage-related integrase and transposase gene의 염기서열을 이용하여 프라이머를 각각 제작하였다. 프라이머 염기서열은 XOP-F (5-CGG TCT GCT CAA TGA GGA AGA-3)와 XOP-R2 (5-TGC AAT TGG TGT TCTCCA GG-3), XOT-F (5-GTC ATA GGT GAG GCT TCCC-3)와 XOT-R2 (5-AGT GCG ATC TTT CAG CAG G-3)로 벼 흰잎마름병균의 DNA를 401bp와 492bp를 증폭하게 제작하였다. PCR 증폭은 벼 흰잎마름병균만 증폭하였으며 다른 세균인 Escherichia coli, Agrobacterim, Pectobacterium caratovora subsp. cartovorum, P. atrosepticum, Pseudomonasputida, P. syringae, P. savastanoi pv. phaeolicola, P. savastanoipv. savastanoi and P. marginalis pv. Marginalis 등은증폭되지 않아 특이성이 인정 되었다. 본 프라이머로 병이 의심되는 벼잎과 논물에서 병원균을 3시간 이내에 검출할 수 있었다.

In order to investigate phenotype and genotype of Salmonella enterica subsp. enterica serovar Enteritidis, Forty-eight S. Enteritidis isolates from diarrhea patients were analysed using antimicrobial resistance typing, Phage typing, and Pulsed field gel electrophoresis in Seoul from 2004 to 2005. All of S. Enteritidis were resistant to streptomycin(SM, 37.5%), ampicillin(AM, 43.8%), t icarcillin(TIC, 43.8%), chloramphenicol(CM, 29.2%), t etracycline(TE, 10 .4%) and nalidixic acid(NA, 18.8%) among 16 antimicrobial drugs. Of 48 S. Enteritidis, 8 isolates(16.7%) were resistant to 1 drug, 3 isolates( 6.3%) to 2 drugs, 1 isolate (2.1%) to 3 drugs and 17 isolates(35.7%) to 4 drugs. The basic pattern of 4 drugs resistance was SM, TIC, TE, and CM but 1 drug resistant isolates represent all nalidixic acid resistance. Among 30 antibiotic r esistant S . Enteritidis, 2 1 isolates(70 %) were phage type 2 1, 8 i solates(26.7%) were phage t ype 2 3 and 1 isolate( 3.3%) was RDNC, respectively. Of the phage types observed, all of phage type 23(8 isolates) were nalidixic acid resistant and phage type 21 were AM-TIC-SM-CM multi-resistance(13 isolates; 43.3%), AM-TIC-SM-TE(4 isolates; 13.3%), AM-TIC-SM(1 isolate; 3.3%), AM-TIC-CM(1 isolate; 3.3%), and AM-TIC(2 isolates; 6.7%) resistance and 1 isolate of RDNC was NA-TE resistance. PFGE divided the isolates into two major clusters, A(n=14) and B(n=14). There were four different resistance profiles with resistance to AM, TIC, SM, TE, NA within PFGE A. Also resistance to AM, TIC, SM, CM was common within PFGE B. The PFGE A strains typed as PT21(n=5), PT23(n=8), and RDNC(n=1), While all the PFGE B strains typed as PT21(n=14). In consequence, there was the highly significant concurrence between resistance typing, phage typing and PFGE.

To investigate antimicrobial susceptibility and relationships of the multidrug resistance and phage types of 49 Salmonella enterica serovar Typhimurium isolated from diarrhea patients in Seoul between 1999 and 2002, we analyed stranis by serotyping, antimicrobial susceptibility test and phage typing. Out of 2,705 samples examined for the causative agent from diarrhea patients, total of 512 isolates were confirmed to be genus Salmonella. Out of 512 isolates, 49 isolates(9.6%) were identified as S. Typhimurium. All of the S. Typhimurium strains represented 100% susceptibility to ceftriaxone, cefoxitin, amikacin and ciprofloxacin and were over 90% susceptibility to gentamicin, cephalothin and kanamycin, but were resistant to tetracycline(75.5%), streptomycin(59.2%), sulfonamides(55.1%), ticarcillin(36.7%), ampicillin( 28.6%), and chloramphenicol(20.4%). Out of 49 S. Typhimurium, only 3 isolates(20.4%) were resistant to one drug, 6 isolates(12.2%) to two drugs and 12 isolates(24.5%) to 3 drugs, 1 isolate(2.0%) to 4 drugs, 2 isolates (4.1%) to 5 drugs, and 6 isolates(12.2%) to 6 drugs and 5 isolates (10.2%) to 7 drugs. Out of 49 S. Typhimurium, 10 isolates(20.4%) were DT195, 5 isolates(10.2%) were DT193 and DT206, 4 isolates(8.2%) were DT104L, DT146, DT203 and RDNC, respectively. Phage types observed the resistant patterns of more than 6 drugs were DT104L, DT193, DT194, DT195 and DT67, but those of 3 drugs representing S-S3-TE type was DT195.

This study was conducted to investigate the effects of different carbon sources on the anaerobic fermentation characteristics in the startup phase using the biochemical methane potential test. The treatments for this experiment were combinations of carbon sources (starch, cellulose, and xylan). Anaerobic fermentation was done at 37oC for 18 days with agitation and pH, ammonia nitrogen, volatile solids reduction, gas production, methane content, and methane production were investigated at 0, 1, 2, 3, 4, 5, 7, 9, 12, 15, and 18 days after incubation in triplicate. In the experiment, the pH was changed depending on the characteristics of the carbon source. The ammonia nitrogen concentration was the highest in the starch-treated group at 7, 12, and 15 days after incubation (P < 0.05). Cumulative volatile solids reduction was the highest in the cellulose-treated group at 18 day after incubation (P < 0.05) and cumulative gas production was higher in the cellulose-treated group than for other two treatments at 18 day after incubation (P < 0.05). Methane content was the lowest in the xylan-treated group at 18 days after incubation (P < 0.05). Cumulative methane production was higher in the xylan-and cellulose-treated group than in the starch-treated group at 18 days after incubation (P < 0.05). In this study, the carbon sources had significant effects on anaerobic fermentation characteristics; especially, the carbon source was shown to have a positive effect on the operation time and hydraulic retention time for the anaerobic digestion startup phage. Therefore, carbon sources should be considered systematically for efficient anaerobic digestion of organic waste.