• Title/Summary/Keyword: E. coli K1 (E44)

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Combined Effect of Cold Plasma and UV-C Against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on Fresh-cut Lettuce (양상추에 인위접종된 Escherichia coli O157:H7, Salmonella Typhimurium과 Listeria monocytogenes에 대한 저온 플라즈마와 UV-C의 살균 효과)

  • Seong, Ji-Yeong;Park, Mi-Jung;Kwon, Ki-Hyun;Oh, Se-Wook
    • Journal of Food Hygiene and Safety
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    • v.32 no.1
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    • pp.64-69
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    • 2017
  • This study was conducted to investigate the effect of cold plasma combined with UV-C irradiation against Escherichia coli O157:H7, Salmonella enterica serovar Typhimurium, and Listeria monocytogenes on lettuce. E. coli O157:H7, S. Typhimurium, and L. monocytogenes, corresponding to approximately 5.82, 5.09, 5.65 log CFU/g, were inoculated on lettuce, respectively. Then, the lettuce was treated with cold plasma, UV-C and combination (cold plasma + UV-C), respectively. The treated lettuce was stored for 9 days at $4^{\circ}C$ for microbiological analysis and sensory evaluation. Cold plasma reduced the populations of E. coli O157:H7, S. Typhimurium, and L. monocytogenes by 0.26, 0.65, and 0.93 log CFU/g, respectively. Each microorganism were reduced by 0.87, 0.88, and 1.14 log CFU/g after UV-C treatment. And, the combined treatment that was treated by cold plasma after UV-C treatment reduced the populations of inoculated microorganisms by 1.44, 2.70, 1.62 log CFU/g, respectively. The all treatment significantly (p < 0.05) reduced the populations of all inoculated bacteria compared to untreated lettuce. UV-C combined with cold plasma was the most effective for reducing the pathogenic bacteria on lettuce, by showing log-reductions of ${\geq}2.0\;log\;CFU/g$. All treatment was not significantly different until 6 day storage compared to control group in terms of appearance, texture and overall acceptability. Therefore, the combined treatment will be an effective intervention method to control the bacteria on lettuce.

Cloning and Expression of the Bacillus thruingiensis var. kurstaki HD-1 Crystal Protein gene in Eschelichia coli

  • Sang Hyn Kim;You
    • Journal of Sericultural and Entomological Science
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    • v.35 no.2
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    • pp.129-133
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    • 1993
  • The 44Md plasmid of Bacillus thruingiensis var. kurstaki HD-1(B. t k HD-1) was partially digested with Sau3AI and the fragments were cloned into E. coli HB101 on vector pBR322. Of 2, 950 clones with a recombinant pBR322, only one clone KC1 was determined to have the gene for crystal toxic proteins from the 44Md plasmid of B. t k HD-1 at the BamHI site of pBR322. The recombinant pBR322 was named pKC1 and its molecular size was 12kb. The KC1 produced a protein which was toxic to the silkworm and antigenically similar to the crystal toxic protein of B. t k HD-1. Also, electrophoretic mobility of the KC1 protein was apparently the same as that of the crystal toxic protein of B. t k HD-1.

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Differential Induction of Septic Shock by Lipopolysacchrides from E. coli and S. abortus (S. abortus 유래 LPS와 E. coli 유래 LPS에 의한 패혈증성 쇽 유도 작용 비교)

  • Cho, Jae-Youl;Yoo, Eun-Sook
    • YAKHAK HOEJI
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    • v.51 no.1
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    • pp.44-50
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    • 2007
  • Acute septic shock is one of inflammatory diseases mediated by pro-inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$. In this study, we examined the pathological difference and mechanism of lipopolysaccharides isolated from E. coli (E-LPS) or S. abortus (S-LPS) on inducing acute septic shock in ICR mouse. All mice were died by intraperitoneal treatment of S-LPS with 0.75 mg/kg, whereas E-LPS treated with even 3 mg/kg only showed 30% of mice lethal, indicating that S-LPS may be more feasible in triggering a strong septic shock condition. The secretion pattern of TNF-${\alpha}$, a critical pro-inflammatory cytokine in septic shock condition, was also distinct between E-LPS- and S-LPS-treated groups. Thus, S-LPS strikingly increased serum level of TNF-${\alpha}$ (6 ng/ml) at 1 h, while E-LPS just displayed at 2 ng/ml level. However the interaction of S-LPS with LPS receptor toll like receptor (TLR)-4, was not stronger than that of E-LPS, according to experiments with macrophage cell line RAW264.7 cells. Thus, E-LPS rather than S-LPS strongly enhanced the production of TNF-${\alpha}$. Interestingly, S-LPS more strongly up-regulated splenocyte proliferation, compared to E-LPS group, whereas there was no difference between S- or E-LPS treated groups in proliferation of Balb/c- or C57BL/6-originated splenic lymphocytes. Therefore, our data suggest that S-LPS is a more active endotoxin and that the strong septic shock-inducing effect of S-LPS seems due to the enhancement of early TNF-${\alpha}$ production and S-LPS-sensitive lymphocyte proliferation.

Antimicrobial Resistance Patterns of Escherichia coli and Vibrio parahaemolyticus Isolated from Shellfish from the West Coast of Korea (서해안 패류에서 분리한 대장균 및 장염비브리오균의 항균제 내성 패턴)

  • Ryu, A Ra;Park, Kunbawui;Kim, Song Hee;Ham, In Tae;Kwon, Ji Young;Kim, Ji Hoe;Yu, Hong Sik;Lee, Hee Jung;Mok, Jong Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.662-668
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    • 2017
  • This study investigated the antimicrobial resistance patterns of Escherichia coli and Vibrio parahaemolyticus isolated from oysters Crassostrea gigas, short-necked clams Ruditapes philippinarum and corb shells Cyclina sinensis from the West Coast of Korea from June through November 2013. The antimicrobial susceptibility patterns of the isolated strains of E. coli and V. parahaemolyticus to 12 antimicrobial agents used in Korea for clinical or veterinary therapy were analyzed. Antimicrobial resistance to at least one antibiotic was seen in 52.0% of the E. coli isolates (156 strains) and 44.3% of the V. parahaemolyticus isolates (194 strains). The resistance of the E. coli (34.0%) and V. parahaemolyticus (41.8%) isolates to ampicillin was highest. Multiple antimicrobial resistance against at least three antimicrobials was seen in 9.0% of the E. coli isolates and 1.0% of the V. parahaemolyticus isolates.

Prevalence of Pathogenic Bacteria in Livestock Manure Compost and Organic Fertilizer (가축분퇴비와 유기질비료에서 병원성박테리아의 분포도 분석)

  • Jung, Kyu-Seok;Heu, Sung-Gi;Roh, Eun-Jung;Lee, Dong-Hwan;Yun, Jong-Chul;Kim, Kye-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.824-829
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    • 2011
  • In recent years, there has been an increasing public concern about fecal contamination of water, air and agricultural produce by pathogens residing in organic fertilizers such as manure, compost and agricultural by-products. Efforts are now being made to control or eliminate the pathogen populations at on-farm level. Development of efficient on-farm strategies to mitigate the potential risk posed by the pathogens requires data about how the pathogens prevail in livestock manure composts and organic fertilizers. Microbiological analysis of livestock manure composts and organic fertilizers obtained from 32 and 28 companies, respectively, were conducted to determine the total aerobic bacteria count, coliforms, Escherichia coli count and the prevalence of Staphylococcus aureus, Bacillus cereus, Salmonella spp., Escherichia coli O157:H7, Listeria monocytogenes, and Cronobacter sakazakii. The total aerobic bacteria counts in the livestock manure composts and organic fertilizers were in the range of 7 to $9log\;CFU\;g^{-1}$ and 4 to $6log\;CFU\;g^{-1}$, respectively. In the livestock manure composts, coliforms and E. coli were detected in samples obtained from 4 and 2 companies, respectively, in the range of 2 to $5log\;CFU\;g^{-1}$ and $2log\;CFU\;g^{-1}$. In the organic fertilizers, coliforms and E. coli were detected in samples obtained from 4 and 1 companies, respectively, in the range of 1 to $3log\;CFU\;g^{-1}$ and $2log\;CFU\;g^{-1}$. In 3 out 32 compost samples, B. cereus was detected, while other pathogens were not detected. In 28 organic fertilizers, no pathogens were detected. The complete composting process can result in the elimination of pathogens in livestock manure compost and organic fertilizer. The results of this study could help to formulate microbiological guidelines for the use of compost in environmental-friendly agriculture. This research provides information regarding microbiological quality of livestock manure compost and organic fertilizer.

Heterologous Production of Streptokinase in Secretory Form in Streptomyces lividans and in Nonsecretory Form in Escherichia coli

  • Kim,, Mi-Ran;Choeng, Yong-Hoon;Chi, Won-Jae;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.132-137
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    • 2010
  • The skc gene encoding streptokinase (SK) with a molecular mass of approximately 47.4 kDa was cloned from Streptococcus equisimilis ATCC 9542 and heterologously overexpressed in Streptomyces lividans TK24 and E. coli using various strong promoters. When the promoter for sprT [Streptomyces griseus trypsin (SGT)] was used in the host S. lividans TK24, a 47.4-kDa protein was detected along with a smaller hydrolyzed protein (44 kDa), suggesting that posttranslational hydrolysis had occurred as has been reported in other expression systems. The casein/plasminogen plate assay revealed that the plasmid construct containing the SGT signal peptide was superior to that containing the SK signal peptide in terms of SK production. Maximal production of SK was calculated to be about 0.25 unit/ml of culture broth, a value that was five times higher than that obtained with other expression systems using ermE and tipA promoters in the same host. When the skc gene was expressed in E. coli BL21(${\Delta}DE3$)pLys under the control of the T7 promoter, a relatively large amount of SK was expressed in soluble form without hydrolysis. SK activity in E. coli/pET28a-$T7_pSK_m$ was more than 2 units/ml of culture broth, even though about half of the expressed protein formed an inactive inclusion body.

Construction of an Escherichia-Pseudomonas Shuttle Vector Containing an Aminoglycoside Phosphotransferase Gene and a lacZ' Gene for $\alpha$-Complementation

  • Lee, Bheong-Uk;Hong, Ja-Heon;Kahng, Hyung-Yeel;Oh, Kye-Heon
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.671-673
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    • 2006
  • A new 4.87 kb Escherichia-Pseudomonas shuttle vector has been constructed by inserting a 1.27 kb DNA fragment with a replication origin of a Pseudomonas plasmid pRO1614 into the 3.6 kb E. coli plasmid pBGS18. This vector, designated pJH1, contains an aminogly-coside phosphotransferase gene (aph) from Tn903, a lacZ' gene for $\alpha$-complementation and a versatile multiple cloning site possessing unique restriction sites for EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, BspMI, PstI, SphI, and HindIII. When pJH1 was transformed into E. coli DHS${\alpha}$ and into P. putida HK-6, it was episomally and stably maintained in both strains. In addition, the enhanced green fluorescent protein (EGFP) gene which was transcriptionally cloned into pJH1 rendered E. coli cells fluorescence when its transformants were illuminated at 488 nm.

Multiplex PCR for Simultaneous Detection of Aminoglycoside Resistance Genes in Escherichia coli and Klebsiella pneumoniae

  • Kim, Hyun Chul;Jang, Ji-Hyun;Kim, Hyogyeong;Kim, Young-Jin;Lee, Kyoung-Ryul;Kim, Yun-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.155-165
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    • 2012
  • The purpose of this study was to develop a multiplex PCR for the detection of aac(6')-Ib, aph(3')-Ia, and ant(2")-Ia; the genes that encode the most clinically relevant aminoglycoside modifying enzymes (AMEs) in Gram-negative bacteria. Clinical isolates of 80 E. coli and 23 K. pneumoniae from tertiary university hospital were tested by multiplex PCR. The most prevalent AME gene was aac(6')-Ib which was found in 22.3% of the isolates. Of the total 80 E. coli isolates, 1 isolate was found to contain both aph(3')-Ia and ant(2")-Ia simultaneouly. Of the total 23 K. pneumoniae isolates, 2 isolates were found to contain both aac(6')-Ib and aph(3')-Ia, and 1 isolate was found to contain both aac(6')-Ib and ant(2")-Ia simultaneously. Annual (2005~2009) analysis of isolates that contain the AME genes were of no correlation. The sensitivity and specificity of multiplex PCR in detecting AME genes was 94.4% (34 of 36 cases) and 100%, respectively. We suggest the multiplex PCR method we developed could be highly sensitive and specific in detecting the AME genes of E. coli and K. pneumoniae. This study could be the first published investigation in which the multiplex PCR method detects aac(6')-Ib, aph(3')-Ia, and ant(2")-Ia genes.

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Evaluation of Potato Varieties (Solanum tuberosum L.) on Fecal Microflora of Human Volunteers

  • Kim, Young-Mi;Lim, Mi-Youn;Lee, Hoi-Seon
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.420-423
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    • 2005
  • Effects of Dasom Valley and Bora Valley on fecal microflora, fecal moisture, and fecal pH of twelve healthy human volunteers were investigated. Numbers of Bifidobacterium, Clostridium perfringens, Escherichia coli, and Lactobacillus of control group were $9.24{\pm}0.63$, $4.44{\pm}1.21$, $7.75{\pm}0.38$, and $6.98{\pm}0.81$ (Log CFU/g wet feces), respectively. During administration of Dasom Valley, numbers of Bifidobacterium and Lactobacillus were $10.70{\pm}0.44$ and $8.84{\pm}0.77$, whereas those of C. perfringens and E. coli were $2.96{\pm}1.50$ and $6.69{\pm}0.29$, respectively. Administration of Dasom Valley significantly increased growth responses of beneficial bacteria, Bifidobacterium and Lactobacillus, whereas those of harmful bacteria, C. perfringens and E. coli, significantly decreased. Moisture content of feces increased and fecal pH decreased with intake of Dasom Valley. Intake of Bora Valley slightly increased numbers of Bifidbacterium and Lactobacillus and slightly decreased those of C. perfringens and E. coli. Results indicate Dasom Valley has greater intestinal-modulating effect than Bora Valley and Atlantic. Daily intake of Dasom Valley may normalize disturbed physiological functions, resulting in improvement of growth and composition of microbial community within intestinal tract.

Comparison of Statistical Methods for Optimization of Salts in Medium for Production of Carboxymethylcellulase of Bacillus amyloliquefaciens DL-3 by a Recombinant E. coli JM109/DL-3 (Bacillus amyloliquefaciens DL-3의 carboxymethylcellulase를 재조합 균주 E. coli JM109/DL-3에서 생산하는 배지의 염 농도를 최적화하기 위한 통계학적 실험 방법의 비교)

  • Lee, You-Jung;Kim, Hye-Jin;Gao, Wa;Chung, Chung-Han;Lee, Jin-Woo
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1205-1213
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    • 2011
  • The optimal concentrations of salts in medium for cell growth and the production of carboxymethylcellulase (CMCase) by a recombinant E. coli JM109/DL-3 were established using two statistical methods: orthogonal array method (OAM) and response surface method (RSM). The analysis of variance (ANOVA) of data based on OAM indicated that $K_2HPO_4$ gave maximum sum of square (S) and percentage contribution (P) for cell growth as well as production of CMCase. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2SO_4$ in medium for cell growth extracted by Qualitek-4 (W32b) Software were 10.0, 1.0, 0.2, and 0.6 g/l, respectively, whereas those for the production of CMCase by E. coli JM109/DL-3 were 5.0, 1.0, 0.4, and 0.6 g/l. The analysis of variance (ANOVA) resulting from RSM indicated that a highly significant salt for cell growth was $K_2HPO_4$ ("probe>F" less than 0.0001), whereas $K_2HPO_4$ and $MgSO_4{\cdot}7H_2O$ were significant for the production of CMCase. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2SO_4$ for cell growth extracted by Design Expert Software were 7.44, 1.08, 0.22, and 0.88 g/l, respectively, whereas those for production of CMCase were 5.84, 0.69, 0.28, and 0.54 g/l. The optimal concentrations of salts and their influences on cell growth and production of CMCase extracted by OAM were almost the same as those by RSM. Production of CMCase by a recombinant E. coli JM109/DL-3 under optimized concentration of salts was 1.93 times higher than that by Bacillus amyloliquifaciens DL-3.