This study investigated viability variation of airborne bacteria in indoor environments. The survival in air as a temporal function of bioaerosol viability was reported for Escherichia coli (KCCM 12119, ATCC 11775). Bacteria suspended in distilled water were aerosolized and entered the vertical duct oriented downward. After measurement of number concentration and colony forming unit (CFU) of the bacteria at different locations of the duct, the viability function was calculated. It was found that the bacteria viability(%) decreased with time after aerosolization, 28.454e^-0.132x (x:time, min). This study demonstrated the potential application of viability function of airborne bacteria to studies of exposure assessment and infection risk analysis.

Recently pathogenic E. coli is one of the main foodborne pathogens resulting in many patients in Korea. To understand the characteristics of pathogenic E. coli outbreaks in Korea, the epidemiological investigation reports of pathogenic E. coli outbreak in 2009 (41 reports) and in 2010 (27 reports) were collected in the web site of the Korea Centers for Disease Control and Prevention, reviewed and analysed in this study. The main places of the pathogenic E. coli outbreaks were food catering service area (64.8%) and restaurants (25.0%). The main type of the pathogens were EPEC (44.7%) and ETEC (34.2%). EAEC and EHEC was responsible for 10.5 and 9.2%, respectively. Eight of 68 outbreak cases were caused by more than 2 types of pathogenic E. coli which implicates the complicated contamination pathways of pathogenic E. coli. The incidence rate of pathogenic E. coli was 33.6 ± 30.5% and the main symptoms were diarrhea, stomach ache, nausea, vomiting, and fever etc. The two identified food sources were identified as frozen hamburger pattie and squid-vegetable mixture. To improve the food source identification by epidemiological investigation, food poisoning notification to the agency should not be delayed, whole food items attributed the outbreak should be collected and detection method of the various pathogenic E. coli in food has to be improved. In conclusion, the characteristics between the EHEC outbreaks in the western countries and the EPEC or ETEC outbreaks in Korea needs to be distinguished to prepare food safety management plan. In addition, the development of the trace back system to find the contamination pathway with the improved detection method in food and systemic and cooperative support by the related agencies are necessary.

The objectives of the current study were to evaluate the therapeutic effect of dioctahedral smectite (smectite) against calf diarrhea caused by pathogenic E. coli and/or Salmonella typhimurium. In this study, 20 calves (aged 2~3 months) with diarrhea were used for evaluation of the efficacy of smectite on calf diarrhea with 20% smectite suspension in PBS. Calves received 10 ml smectite suspension three times per day after feeding, and fecal samples were collected at the gate of treatment and on the first, second, third, fourth, and fifth day after administration. On the fifth day after treatment with smectite suspension, the diarrhea index showed a significant decrease in the treated group, compared to the control group (P<0.001). The number of pathogenic E. coli in feces of the treated group was significantly decreased, compared to each control group from the second day after treatment (P<0.001), and that of Salmonella typhimurium was significantly decreased from the first day after treatment (P<0.05). According to the results of the current study, 20% smectite suspension had a therapeutic effect on diarrhea caused by E. coli and/or Salmonella in calves.

Hemorrhagic Escherichia coli (E. coli) is one of the most important agent of diarrhea in piglet. The aim of this study was to investigate the characteristics of virulence genes and genetic diversity in hemorrhagic E. coli isolated from piglets with diarrhea. Among 122 hemorrhagic E. coli, 62 isolates carried single toxin gene like heat-stable toxin (ST), heatlabile toxin (LT), verotoxin 1 (VT1), verotoxin 2 (VT2), spa and eae. The most prevalent toxin gene carried by isolates was ST gene (23 isolates), while the most common association was ST/LT (14 isolates) and ST/LT/VT2 (13 isolates). In pulsed field gel electrophoresis, isolates were not classified as one cluster by epidemiological information including isolated area, isolated year and possessed toxins.

To determine current rate of antimicrobial resistance, a total of 236 isolates from milk samples of dairy cattle with mastitis in Korea during 2010-2011 were examined against 12 antimicrobials using disc diffusion method: 67 Staphylococcus aureus, 74 coagulase-negative Staphylococcus spp. (CNS), and 95 Escherichia coli isolates. The isolates examined in this study were submitted by Local Veterinary Service Laboratories located in 13 provinces and metropolitan cities nationwide. The highest rates of resistance among S. aureus isolates were against ampicillin (56.7%) and penicillin (56.7%), followed by kanamycin (11.9%). All S. aureus isolates were sensitive to lincomycin, amikacin, and cephalothin. Only one isolate showed resistance to tetracycline and oxacillin, respectively. Less than 10% of the S. aureus isolates presented resistance to erythromycin, neomycin, and gentamicin. Among CNS isolates, the most frequently observed resistance was to lincomycin (44.5%), followed by penicillin (28.3%), ampicillin (18.9%), tetracycline (17.5%), kanamycin (13.5%), and erythromycin (9.4%). All or most of the CNS isolates were sensitive to cephalothin, amikacin, neomycin, and gentamicin. The highest rate of resistance among E. coli isolates was against tetracycline (26.3%), followed by streptomycin (21%), neomycin (15%), kanamycin (12.6%), and gentamicin (10.5%). Amikacin was the only antimicrobial to which no E. coli isolates showed resistance. Around 10% of the S. aureus isolates and 15% of the CNS isolates showed resistance against three or more antimicrobials simultaneously, while more than 30% of the E. coli isolates did.

서울시내에서 유통되는 식품과 식품접객업소(집단급식소 포함)의 조리식품을 대상으로 식중독 원인균 분석 및 위생미생물 검사를 실시한 결과, 분리된 대장균의 항생제 감수성 시험을 통하여 이들의 내성 정도를 파악하고, 내성 유전자와 병원성유전자의 분포도 알아보았다. 모두 1313건 의 샘플 중 50건에서 대장균이 검출되어 3.8%의 검출률을 보였다. 이중 육회 1건에서 장출혈성대장균 O26 1건, 김밥 에서 장병원성대장균 1건이 각각 검출되었다. 50건의 대장 균중 50%가 16종의 항생제에 모두 감수성을 보였으며 내성이 높게 나타난 항생제는 ampicillin(36%), amoxicillin/ clavulanic acid(32%) 그리고 tetracycline(22%)의 순이었다. 이들의 내성유전자 분포는 TEM이 1건, tetB 4건이 각각 검출되었다.

Bacillus thuringiensis 1-3 (Bt 1-3), belonging to subsp. aizawai (H7), showed different characteristics in plasmid profiles from type strain and had cry2A gene in addition to cry1Aa, cry1Ab, cry1C and cry1D. To clone its plasmids and construct E.coli-Bt shuttle vector, we constructed the plasmid capture system (PCS) by inserting attB sites including lacZ between transposable elements (designated as pPCS-Troy). Through in vitro transposition with total plasmids DNA of Bt 1-3, 53 clones were acquired and their sizes were approximately 10 kb. Based on the sequence analysis, they were classified in four groups showing similarities with four known Bt plasmids, pGI3, pBMB175, pGI1 and pGI2, respectively. One of pGI3-like clones, named as pBt1-3, was fully sequenced and its putative open reading frames (ORFs), Rep-protein, double-strand origin of replication (dso), single-strand origin of replication (sso), have been identified. The structure of pBt1-3 showed high similarity with pGI3 which is one of rolling-circle replication (RCR) group VI family. As a donor for construction of shuttle vector, pDonr-attPEm vector harboring erythromycin resistant gene between attP sites was constructed. Through BP recombination with pPCS-Troy-cloned Bt plasmids and pDonr-attPEm, erythromycin resistant gene was transposed to Bt plasmids. This scheme proposes that in vitro transposition using pPCS-Troy and BP recombination using pDonr-attPEm can easily clone Bt plasmids and construct novel shuttle vectors.

The desmutagenic activity of the water extract of Cassia tora L on the mutagenicity induced by 4- nitroquinoline 1-oxide (4-NQO) and N-methyl-N-nitro-N'-nitrosoguani-dine (MNNG) was studied using the SOS Chromotest with Escherichia coli PQ37. The inhibition rates of water extract of Cassia tora L. at concentration of 100 μg/assay were 27.5% and 40% against 4-NQO and MNNG. The water extract of Cassia tora L. was separated into methanol soluble and methanol insoluble parts. The methanol soluble part exhibited higher inhibition effects than the methanol insoluble part against the mutagenic activities of 4-NQO and MNNG. Step-wise fractionation of methanol soluble part was done to obtain methanol, ethyl acetate and water fractions. Among these fractions, water fraction had the strongest inhibitory effects of 23.0 and 19.0% against mutagenicities of MNNG and 4NQO, repectively. The results clearly indicated that the water fraction showed much stronger antimutagenicity against MNNG than 4NQO. The inhibition rates of aqueous fraction of methanol-soluble from water extracted Cassia tora L. at concentrations of 1.0 10, 100 and 250 μg/assay were 8.0%, 12.0%, 25.5% and 43.0%, respectively. The water fraction showed the inhibitory effects with dose response against the mutagenic activities induced by MNNG.

Carbon Nano Tubes could be either metallic or semi-conducting in nature, depending on their diameter. Its photocatalytic behavior has given an impetus to use it as an anti-microbial agent. More than 95% Escherichia coli and Staphylococcus aureus bacteria got killed when exposed to Carbon Nano Tubes for 30 minutes in presence of sunlight. Carbon Nano Tubes are supposed to have smooth surface on to which it accumulates positive charges when exposed to light. The surface that is non illuminated has negative charge. At the cellular level microorganisms produce negative charges on the cell membrane, Therefore damaging effect of multi walled carbon nano tubes (exposed to light) on the microorganisms is possible. In this paper, photo catalytic killing of microbes by multi walled carbon nano tubes is reported. Killing was due to damage in the cell membrane, as seen in SEM micrographs. Moreover biochemical analysis of membrane as well as total cellular proteins by SDS PAGE showed that there was denaturation of membrane proteins as well as total proteins of both the microbes studied. The killed microbes that showed a decrease in number of protein bands (i.e. due to breaking down of proteins) also showed an increase in level of free amino acids in microbes. This further confirmed that proteins got denatured or broken down into shorter units of amino acids. Increased level of free amino acids was recorded in both the microbes treated with multi walled carbon nano tubes and sunlight.

본 연구는 시판되는 증자 녹차의 에탄올 추출물과 에탄올 추출물로부터 분획한 분획물로부터 최근 병원성 식중독균으로서 문제시되고 있는 E. coli O157:H7균에 대한 항균성을 검색하였다. 에탄올 추출물로부터 그 유효성분을 조사하기 위하여 핵산, 에틸아세테이트, 클로로포름 및 물로서 분획한 후 MIC를 검색한 결과 에틸아세테이트 분획물이 500μg/disk 농도로 가장 강한 활성을 나타내었으며, 헥산 분획물은 활성이 없었다. 가장 강한 항균력을 나타낸 에틸아세테이트 분획물의 농도별 항균력 및 pH에 대한 영향을 조사한 결과 균종에 따라 차이는 있지만 3균주 모두 250-2000μg/disk 농도에서 새육이 억제되었다. 또한 E. coli O157:H7균은 산성조건에서는 생육이 가능하지만 pH 10 이상의 강한 알칼리 조건에서는 생육이 크게 억제되었다. 35℃에서 E. coli O157:H7 933과 932균에 대한 에틸아세테이트 분획물의 농도별 생육저해 효과를 검색한 결과 두 균주 모두 250μg/ml , 500μg/ml의 농도에서는 12시간 이후부터 생육하기 시작하였으나 1000μg/ml의 농도에서는 생육이 크게 저해되었다.

식품으로부터 다양한 병원 미생물을 신속 검출하기 위하여 다양한 검출 원리를 이용한 키트들이 개발·시판되고 있다. 검사키트는 신속, 정확하고 간단하게 사용할 수 있으므로 검사기관이나 실험실 뿐 아니라 식품회사에서 QC 또는 QA를 수행하기 위하여 사용이 증가되고 있는 추세이다. 이에 본 연구에서 E. coli O157:H7의 단클론항체를 이용하여 면역크로마토그래피법에 의해 개발한 E. coli O157:H7 검출 키트(Donga Co, Korea, D-kit)에 대한 검출감도 및 특이성을 확인하고 식품 시료에 적용 가능성을 평가하였다. 면역크로마토그래피법에 의해 개발?시판되고 있는 Reveal E. coli O157:H7 (Neogen Co., USAm R-kit)와 VIP EHEC kit (Biocontrol INC., USA, V-kit)를 비교 키트로 사용하였다. E. coli O157:H7 표준균주를 사용하여 실시한 검출감도 확인시험 결과 R-kit 및 D-kit는 104/㎖의 농도에서 양성으로 확인되었고 10³/㎖에서도 약한 양성 반응을 보였으나, V-kit는 10?/㎖ 농도로 검출감도가 낮았다. 또한 배양액을 가열하여 kit에 적용하는 것이 가열하지 않은 경우보다 검출감도를 높일 수 있었다. E. coli O157:H7 분리 22주, verotoxin 생성 E. coli 7주 E. coli 분리주 40주 및 38종의 장내세균에 대한 특이성 시험 결과 세 가지 키트 모두에서 동일한 결과를 보였는데, 시험균주 107주 중 3주를 제외한 모든 균에서 음성의 결과를 보여 특이성을 확인하였다. 세 키트에 위양성 반응을 보인 것은 E. coli O157:H19, E. coli O157:H18 및 Salmonella galinarium으로 이들 혈청형과 O157:H7 사이에는 유사한 혈청학적 특성이 존재하는 것으로 추정되었다. 이상의 실험결과로 D-kit는 E. coli O157:H7을 검출하는데 감도 및 특이성 면에서 기존 키트인 R-kit 및 V-kit와 같이 유용한 것으로 확인되었다.

In the 5'-flanking region of the D-AAT, AspAT and AlaDH gene, I found three or two pairs of sequences(designated as P1, P2, P3) which show significant similarity to the E.coli consensus sequences of -35 and -10 for promoters. The spacing between -35 and -10 is 16 to 18bp in all the three putative promoters Pl, P2 and P3 which is in good agreement with the preferred spacer length in E.coli and in B.subtilis. Therefore, the putative promoters may also function to increase the efficiency of transcriptional initiation. The most stable, double-helical $quot;Shine-Dalgarno$quot; pairing is formed with a free energy change(△G) of -13.0 ㎉/㏖, -9.6 ㎉/㏖, -15.8 ㎉/㏖.

The in vitro effects of trisodium phosphate and cetylpyridinium chloride on E. coli O157:H7 and L. monocytogenes were investigated. The trisodium phosphate and cetylpyridinium chloride was bactericidal toward E. coli O157:H7 and L. monocytogenes. The killing effects of the 1 × 10^(-2) M trisodium phosphate on E. coli O157:H7 and L. monocytogenes were 30-40%, 40-50%, respectively. The killing effects of the 5 × 10^(-7) M cetylpyridinium chloride on E. coli 0157:H7 and L. monocytogenes were 90-95%, 95-99%, respectively. The killing effects of the trisodium phosphate was 105 times that of the cetylpyridinium chloride. Factors effecting the bactericidal action of trisodium phosphate and cetylpyridinium chloride were investigated and the action depended on temperature and pH.

최근 산업의 발달로 인한 식품의 안전성에 대한 국민의 우려가 증가되고 있으며 특히 식품을 미생물의 증식으로부터 안전하게 보존하기 위한 천연식품보존제의 개발이 요구되고 있다. 지구상에 풍부한 천연자원인 키토산과 합성화학보존제인 소르빈산을 사용하여 그람음성 병원성 식중독 원인균인 E. coli O157:H7과 대표적 그람양성 식중독 원인균인 Staph. aureus에 대해 실험한 결과는 다음과 같았다. 5. coli O157:H7에 대한 키토산의 최소발육억제농도는 pH 5.0에서 500 ppm, pH 5.5에서 250 ppm, pH 6.0에서 500 ppm, 그리고 pH 6.5에서 2000 ppm이었으며, Staph. aureus에 대한 키토산의 최소발육억제농도는 pH 5.0에서 31.3 ppm이었으며 , pH 5.5 이상에서는 62.5 ppm이었다. 소르빈산의 최소발육억제농도는 pH 5.0에서 500 ppm, pH 5.5에서 1500 ppm, 그리고 PH 6.0 이상에서는 2000ppm이상이었다. Staph. aureus에 대한 소르빈산의 최소발육억제농도는 pH 5.0에서 1500ppm이었으며, pH 5.5 이상에서는 2000 ppm 이상이었다. E. coli O157:H7에 대한 키토산과 소르빈산의 병 용처리효과는 pH에 크게 영향을 받아 pH 6.5에서는 키토산농도 500 ppm과 소르빈산 500 ppm농도에 발육이 억제되었으나 pH 5.0에서는 키토산농도 250 ppm과 소르빈산 31.3 ppm에 억제되었다. 한편, Staph. aureus에서는 pH에 대한 영향이 별로 없었으며, 키토산의 영향을 크게 받으나 소르빈산의 영향은 거의 없었다. pH 6.5, 37℃의 조건하에서 E. coli O137:H7은 키토산 500 ppm 및 소르빈산 500 ppm 처리와 키토산 250 ppm 및 소르빈산 250 ppm 병용처리군에서 모두 증식이 억제되었으나 키토산과 소르빈산 단독처리군에서는 배양시간이 증가함에 따라 균의 증식이 계속되었다. Staph. aureus는 소르빈산에 영향을 받지않았으며 병용효과보다는 키토산에 대한 영향이 컸다. pH 5.5, 37℃의 조건하에서 E. coli O157:H7은 키토산 500 ppm, 250 ppm 단독첨가시와 병용처리시 모두 3시간 이내에 균증식이 억제되었다. Staph. aureus의 경우는 키토산 50 ppm 단독처리군과 키토산 25 ppm과 소르빈산 2000 ppm 병용처리군에서 균증식이 사면되었다. E. coli O157:H7에 대한 키토산과 소르빈산병용처리시 MIC와의 상관관계는 pH 6.5에서 R=0.95(p<0.01), pH 6.0에서 R=0.99(p<0.01), pH 5.5에서 R=0.97(p<0.01), 그리고 pH 5.0에서 R=0.99(p.0.01)로 높은 상관관계를 나타내었다. Staph. aureus에 대해서는 소르빈산의 병용처리에 의한 효과는 거의 없었다. 이상의 결과로 종합하면 pH변화에 따른 키토산과 소르빈산 단독 및 병용처리로 인한 상승효과를 확인할 수 있었으며 이 결과 천연식품인 키토산을 병용함으로써 인체에 영향이 있는 합성화학보존제인 소르빈산의 양을 감소시킬 수 있었으며, 또한 식품의 보존성을 더 증가 또는 유지시킬수 있을 것으로 기대되었다.

A study was conducted to determine contamination status of fish sold at wholesale market in Seoul. A total of 79 samples (35 different kindry fish) were collected from the wholesale market. E. coli and coliform group bacteria were cultured and tested for sensitivity against antibiotics. The results are summarized as follows; 1. E. coli was isolated from 23 out of 79 samples (29.1%), and coliform groups from 53 out of 79 (67.1%). 2. Of coliform group, Citrobacter freundii was the most common and Enterobacter cloacae was the next. 3. 23 E. coli strains isolated from fishes were resistant to Oxacillin, Erythromycin and Lincomycin, meanwhile 23 E. coli strains were sensitive to Cefoperazone, Ceftazidime, Imipenem, and Ciprofloxacin.

The growth inhibition effect of orally administrated yogurt ACE and Metchnikoff upon E. coli O157:H7 and S. typhimurium inoculated into gastric lumen of rabbits was investigated. The rabbits challenged with each 1 ml of suspension containing 10^8 CFU/ml of the pathogens were divided into 4 groups by the interval of yogurt administration: A group; preadministrated 7 days before inoculation of the pathogens and fed daily; B group; administrated daily after inoculation of the pathogens, C group; administrated every 3 days after inoculation of the pathogens; Control group, not fed after inoculation of the pathogens. Each 3 ml of yogurt containing 10^9 CFU/ml was orally administrated into rabbits. All yogurt administrated groups (A, B, C) showed growth inhibition effect on E. coli O157:H7 in one day after inoculation of the pathogen by the level of 0.8-1.0 log CFU/g, compared with the result differences between the control group and the yogurt administrated groups. In the control group after 5 days of inoculation, the number of colonized pathogens was 10^5-10^6 CFU/g, whereas 10³-10⁴ CFU/g was detected in the yogurt administrated groups. After 10 days of inoculation, the viable pathogen number per gram (g) of the rabbit feces was 10³ CFU/g in the control group, whereas the number below 10¹ CFU/g was detected in the group A, and 10² CFU/g in the group B and C. The growth inhibition effect of yogurt administration on E. coli O157:H7 was highly increased in the order of A, B, and C group. The same effect on S. typhimurium was observed at the level of 2 log CFU/g in the Metchnikoff yogurt administrated groups, compared with the control group result in one day after inoculation of the pathogen. In 7 days after inoculation of the pathogen, the viable number was increasingly decreased, and finally after 15 days no viable cell of S. typhimurium was discharged into the fecal samples in the group A, and the mean level of 10¹ CFU/g was detected in the group B, but there was no growth inhibition effect in the group C. The growth inhibition effect on S. typhimurium was observed at the same level of viable cell number between the yogurt ACE administrated groups and the control group in 5 days after inoculation. But, after 10 days of inoculation the viable cell number was started to decrease, and the viable cell of S. typhimurium was not discharged from rabbit intestinal contents after 15 days of inoculation in the yogurt ACE administrated groups. In such a case that yogurt was administrated in order to prevent the pathogens, pre-administration on a daily basis one week before inoculation of the pathogens exerted considerable effect in growth inhibition. In comparison with two kinds of yogurt tested in this study, the growth inhibition effect on two kinds of pathogens was observed more highly in the Metchnikoff administrated group than the ACE administrated group.

Lactobacillus acidophilus NCFM, Lactobacillus casei YIT 9018, Bifidobacterium longum 8001, and Bifadobacterium longum 8025 at the level of 10^6 cfu/ml were cultured with 10⁴ cfu/ml of Escherichia colt O157:H7 KSC 109 or Salmonella typhimurium ATCC 14028, in order to verify the effects of lactic acid bacteria and bifidobacteria on the growth of the pathogens. In the mixed culture of lactic acid bacteria with E. colt O157:H7 KSC 109, growth inhibition and atypical microcolonies of E. colt O157:H7 KSC 109 were observed. The pathogens inoculated grew for 5 hours (pH 5.3), by the time L. acidophilus NCFM reached the exponential growth phase, and then the surviving pathogens were decreased to 10¹ cfu/ml after 35 hours. When L. casei YIT 9018 was grown with the pathogens, they grew for 10 hours (pH 4.6), by the time L. casei YIT 9018 reached the end of exponential growth phase, and then the surviving pathogens were decreased drastically. Up to the stationary growth phase of lactic acid bacteria, L. acidophilus NCFM exhibited stronger inhibition against the pathogens than L. casei YIT 9018 did, which might be attributed to its faster growth. Likewise bifidobacteria inhibited the growth of the pathogens. tested, bifidobaceria was weaker in the inhibitory activity than lactic acid bacteria. When Bifidobacterium longum 8001 was cultured with the pathogens, E. colt O157:H7 KSC 109 was gradually inhibited at the stationary growth phase of bifidobacteria, atypical microcolonies were formed on Levine EMB medium after 48 hours, and Salmonella grew up to 10^6 cfu/ml, then was drastically inhibited at the exponential growth phage of Bifxdobacterium longum 8001. But when Bifidobacteriuam longum 8025 was cultured with the pathogens, the pathogens grew to the same level of Bifidobacteriuam longum 8025 after 10 hours, then the surviving pathogens were decreased drastically.