• Title/Summary/Keyword: toxin production

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Analysis of Transgenic Mouse, for the Production of Immunodeficiency Animals (면역결핍동물의 생산을 위한 형질전환생쥐의 분석)

  • 나루세겐지;양정희;이승현;최화식;이성호;박창식;진동일
    • Korean Journal of Animal Reproduction
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    • v.27 no.2
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    • pp.179-185
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    • 2003
  • To determine whether the diphtheria toxin-A (DT) gene disrupts development of thymocytes in transgenic animal, the DT-A gene was used for the production of transgenic mice directed by proximal Ick promoter sequences. Two transgenic founder mice that contained several copies of transgene were produced by DNA microinjection and integration of transgene in transgenic mice was confirmed by PCR and Southern blotting analysis. Transgenic $F_1$ and $F_2$ mice were produced by outbreeding of founder and $F_1$ mice to investigate expression of transgene and phenotypes in transgneic mice. Expression of the diphtheria toxin gene was confirmed in thymus, spleen and liver of transgenic mice by RT-PCR. In circulating blood of transgenic mice, lower number of circulating white blood cells and platelets were observed compared with that of normal mice. In addition, transgneic mice had reduced number of circulating peripheral T-cells analyzed by FACS with anti-CD3 antibody. The data in these transgenic mice indicate that DT gene can play a disruptive role in developing thymocytes of transgenic mice resulted in lower number of T-cells that can be applicable to a wide range of tissues in other animals.

Production of 8-ketotrichothecenes by Fusarium graminearum on Corn and Barley (옥수수와 보리에서 Fusarium graminearum의 8-ketotrichothecenes 생성)

  • 서영수;서정아;손황배;이인원
    • Korean Journal Plant Pathology
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    • v.14 no.5
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    • pp.418-424
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    • 1998
  • The production of 8-ketotrichothecenes, deoxynivalenol (DON), nivalenol (NIV), and their monoacetyl derivatives was studied in rice and corn cultures using 8 isolates of Fusarium graminearum which were obtained from corn and barley samples. Higher concentrations of trichothecenes were produced on rice than corn, and production of the toxins on rice was enhanced by growing the fungi at $25^{\circ}C$. The isolates were used for evaluation of toxin production and pathogenicity after artificial inoculation to 5 corn and 3 barley cultivars. The kinds and the relative amounts of trichothecenes produced in cultures were consistent with those in infected kernels of corn and barley with some exceptions. As for DON chemotypes, the ratios of 15-acetyl-DON to 3-acetyl-DON were varied among the pathogen-cultivar interactions. The corn and barley cultivars showed the significant differences of resistance to the Fusarium isolates in disease severity and seedling blight, and resistance ranking to the different isolates was varied. However, significant correlations were observed between the total concentrations of trichothecenes in infected kernels of corn and barley and pathogenicities of the Fusarium isolates to the hosts.

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Relationship between Gb3 Expression and Cytotoxicity of Shiga-like Toxin I (Shiga-like Toxin I의 세포독성과 수용체 Gb3 발현과의 관계)

  • Lim, Suk-Hwan;Kim, Gi-Young;Kim, Hyung-Chun;Kim, Young-Hee;Son, Yong-Hae;Oh, Yang-Hyo;Park, Yeong-Min
    • Clinical and Experimental Pediatrics
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    • v.46 no.2
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    • pp.143-153
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    • 2003
  • Purpose : Infection with Shiga-like toxin (SLT)-producing Escherichia coli, an emerging human pathogen found particularly in young children under 5 years of age, causes a spectrum of illnesses with high morbidity and mortality, ranging from diarrhea to hemorrhagic colitis and hemolytic uremic syndrome. Host mediators play an important role in the pathogenesis of SLT-I toxicity. The experiments described here were designed to investigate the effect of SLT-I on TNF-${\alpha}$ production and to understand the effect of TNF-${\alpha}$ on GB3 expression. We also further examine the relationship between the Gb3 level and the differential susceptibility of cells to the cytotoxic action of SLT-I. Methods : The effect of purified SLT-1 from E. coli O157 : H7 (ATCC 43890) on tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) production in Raw264.7 cells was investigated. Many mediators regulate endothelial cell membrane expression of the glycolipid globotriaosyleramide (Gb3), which serves as the toxin receptor, suggesting that the host response to the toxin or other bacterial products may contribute to pathogenesis by regulating target cell sensitivity to the toxins. Therefore, the relationships between Gb3 expression and cytotoxicity against SLT-I on three types of cells were evaluated. Results : Detectable levels of TNF-${\alpha}$ were produced as early as six hours after induction and continued to increase during 48 hours by SLT-I. It was also found that Vero cells and dendritic cells (DC2.4 cells) expressed high levels of Gb3, 83% and 68%, respectively, and that Raw264.7 cells had a low level of Gb3 (29%) and appeared refractory to cytotoxicity against SLT-I. Vero cells and DC2.4 cells expressing high levels of Gb3 were highly susceptible to SLT-I. Furthermore, macrophages showed a resistance to SLT-I cytotoxicity, despite the fact that Gb3 expression was enhanced. Conclusion : These results strongly suggest that the expression of Gb3 is necessary but not sufficient to confer sensitivity of macrophages to SLT-I and further underpin the important role of SLT-I and its Gb3 receptors in the pathogenesis of E. coli O157 infection.

Transfer of Genes for Antimicrobial Resistance and Toxin of Hemolytic Escherichia coli Isolated from Feces of Pig Suffering Diarrhea to Human Isolates (설사 증상의 돼지 분변에서 분리된 용혈성 대장균의 항생제 내성과 독소의 인체로부터 분리된 균주로의 전이)

  • Lee Kyenam;Jung Byeong Yeal;Lee Yeonhee
    • Korean Journal of Microbiology
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    • v.40 no.4
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    • pp.286-294
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    • 2004
  • Between 1997 and 1998 in Korea, 56 isolates of Escherichia coli were obtained from pig suffering diarrhea. Among those, 38 isolates that showed the hemolytic activity, antimicrobial resistance, and toxin production were studied. Among 38 isolates, thirty-six isolates $(94.7\%)$ were resistant to tetracycline, 27 isolates $(71.0\%)$ were resistant to ampicillin, 26 isolates $(68.4\%)$ were resistant to chloramphenicol, and 21 isolates $(55.2\%)$ were resistant to trimethoprim, while none was resistant to aztreonam, amikacin, and norfloxacin. Among these iso­lates, 21 isolates $(55.3\%)$ were multiple drug resistant to at least four different class antimicrobial agents. Extended spectrum $\beta-lactamase$ producing isolates were not detected in the double disk synergy test. In these hemolytic Escherichia coli, heat-stable enterotoxin $(89.5\%)$ was the most prevalent toxin, followed by vero­toxins $(47.4\%),$ and then heat-labile enterotoxin $(31.6\%).$ Except 8 isolates $(21.0\%)$ which produced ST only, 12 isolates $(31.6\%)$ produced ST and LT, 13 isolates $(34.2\%)$ produced ST, VT, and VTe, and 5 isolates $(13.2\%)$ produced VT and VTe. However, none produced all 4 types of toxin, simultaneously. The predominant serotype could not be determined by the agglutination method. Sixteen isolates $(42.1\%)$ were strongly adhered to T-24 bladder cell and 17 isolates $(44.7\%)$ were to Caco-2 intestinal cell. Especially, 11 strains $(28.9\%)$ were evaluated as strongly adhesive to both T-24 cells and Caco-2 cells. Genes for toxin and the antimicrobial resistance were transferred to clinical isolates of Escherichia coli from human urine by the filter mating method. Results suggest the possibility that antimicrobial resistance and toxin can be transferred from animals to humans by direct con­tact of resistant bacteria as well as gene transfer, although there was no correlation between toxin production, adherent activity, and antimicrobial resistance among hemolytic E. coli isolated from pig suffering diarrhea.

P2 Receptor-mediated Inhibition of Vasopressin-stimulated Fluid Transport and cAMP Responses in AQP2-transfected MDCK Cells

  • Kim, Yang-Hoo;Choi, Young-Jin;Bae, Hae-Rahn;Woo, Jae-Suk
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.1
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    • pp.9-14
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    • 2009
  • We cultured canine kidney(MDCK) cells stably expressing aquaporin-2(AQP2) on collagen-coated permeable membrane filters and examined the effect of extracellular ATP on arginine vasopressin(AVP)-stimulated fluid transport and cAMP production. Exposure of cell monolayers to basolateral AVP resulted in stimulation of apical to basolateral net fluid transport driven by osmotic gradient which was formed by addition of 500 mM mannitol to basolateral bathing solution. Pre-exposure of the basolateral surface of cell monolayers to ATP(100 ${\mu}M$) for 30 min significantly inhibited the AVP-stimulated net fluid transport. In these cells, AVP-stimulated cAMP production was suppressed as well. Profile of the effects of different nucleotides suggested that the $P2Y_2$ receptor is involved in the action of ATP. ATP inhibited the effect of isoproterenol as well, but not that of forskolin to stimulate cAMP production. The inhibitory effect of ATP on AVP-stimulated fluid movement was attenuated by a protein kinase C inhibitor, calphostin C or pertussis toxin. These results suggest that prolonged activation of the P2 receptors inhibits AVP-stimulated fluid transport and cAMP responses in AQP2 transfected MDCK cells. Depressed responsiveness of the adenylyl cyclase by PKC-mediated modification of the pertussis-toxin sensitive $G_i$ protein seems to be the underlyihng mechanism.

Prevalence and Toxin Genes of Food-Borne Pathogens Isolated from Toothbrush in Child Care Center (보육시설 유아 사용 칫솔의 식중독 미생물 분포 및 독소 유전자)

  • Kim, Jong-Seung;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.242-248
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    • 2015
  • This study was performed to investigate the microbiological contamination on toothbrushes, toothbrush caps, and tooth cleaning cups in the child care centers and to evaluate the toxin genes, toxin production ability and antibiotic resistance of food-borne pathogens. The average number of total aerobic bacteria and fungi were 5.3 log CFU and 3.2 log CFU. Coliform bacteria were detected in 41 (54.7%) of 75 toothbrushes, 13 (44.8%) of 29 toothbrush caps, and 29 (44.6%) of 65 tooth cleaning cups. Salmonella spp. was not detected in all of samples but Bacillus cereus was isolated from 1 (1.3%) of 75 toothbrushes and 2 (3.1%) of 65 tooth cleaning cups. Staphylococcus aureus was detected in 1 (1.5%) of 65 tooth cleaning cups. The nheA, nheB, nheC, hblC, hblD, hblA and entFM toxin genes were possessed in B. cereus isolated from toothbrush which also produce NHE and HBL enterotoxins. S. aureus was resistant to ampicillin and penicillin, while B. cereus was resistant to ${\beta}-lactam$ antibiotics. These results indicated that the sanitary conditions of toothbrushes and tooth cleaning cups in the child care centers should be improved promptly. The UV sterilization after drying and then storage in dried condition is required to improve the sanitary condition of toothbrushes and tooth cleaning cups in the child care center.

Effect of Polyphosphates on the Growth of Penicillium griseofulvum and the Production of Patulin (Penicillium griseofulvum 성장과 Patulin 생성에 미치는 인산염의 효과)

  • 김승교;강성조;송재영;전향숙;강진순;김일환;정덕화
    • Journal of Environmental Health Sciences
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    • v.21 no.3
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    • pp.23-27
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    • 1995
  • To extend the shelf lives of rice and corn products, the effects of the polyphosphates[$Na(PO_3)n$, n=11] on the growth of Penicillium griseofulvum and patulin production were investigated. The growth was completely inhibited in the potatoes dextrose agar medium treated with 2% polyphosphate. Moisture content had a considerable influence on the production of patulin. At 30% moisture content, the amounts of patulin produced in rice and corn were $61.40 \mu g/ml$ and $40.74 \mu g/ml$, respectively, but the level of the toxin was significantly decreased to 93~95% by addition of 1% polyphosphates. No patulin was detected in both rice and corn medium added 2% polyphosphate when the incubation time prolonged. The result of scanning electron microscopy was supposed that the biocidal action of polyphosphate on fungi was related to the collapse of cell wall structure.

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Cytochalasin E Production by Rosellinia necatrix and Its Pathogenicity on Apple (사과흰날개무늬병균의 Cytochalasin E 독소 생산과 병원성)

  • Lee, Dong-Hyuk;Choi, Kyung-Hee;Uhm, Jae-Youl
    • Research in Plant Disease
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    • v.15 no.1
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    • pp.46-50
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    • 2009
  • Cytochalasin E (CE) is a secondary metabolite secreted by Rosellinia necatrix, caused by white root rot, and has toxicity to apple as a toxin during disease progress. This study was conducted to demonstrate the relationship between the production of CE and its pathogenicity. CE producing isolates and non-producing isolates of R. nectatrix were isolated from the mycerial mat of diseased roots and was detected on that using a TLC and HPLC analysis and in vivo pathogenicity test. CE non-producing isolates were not pathogenic to apple roots and not detected CE by TLC and HPLC analysis. It was shown that the production of CE was related to the pathogenicity of R. nectatrix.

Detection, Characterization and Antibiotic Susceptibility of Clostridioides (Clostridium) difficile in Meat Products

  • Muratoglu, Karlo;Akkaya, Esra;Hampikyan, Hamparsun;Bingol, Enver Baris;Cetin, Omer;Colak, Hilal
    • Food Science of Animal Resources
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    • v.40 no.4
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    • pp.578-587
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    • 2020
  • Clostridioides (Clostridium) difficile is a Gram (+), anaerobic, spore forming, rod shaped bacterium that can produce toxin. The objective of this study is to reveal the presence of C. difficile in meat products, to analyze the ribotype diversity by PCR and to evaluate the antibiotic susceptibility of isolated strains. The organism was isolated in 22 out of 319 (6.9%) examined meat product samples and 9 out of 22 (40.9%) isolates were identified as RT027 and all isolates had the ability of toxin production. In terms of antibiotic susceptibility, all isolates were susceptive to amoxicillin-clavulanic acid, tetracycline and vancomycin and 21 (95.4%) isolates to metronidazole. On the other hand, imipenem and cefotaxim resistance was observed in all. In conclusion, the results of this comprehensive study conducted in Turkey deduced the presence of C. difficile in different meat products. Therefore, these products can be evaluated as a potential contamination source of C. difficile from animals to humans especially for elders, youngsters, long terms wide spectrum antibiotic used and immuno-suppressed individuals.

A Technique to Enhance Bacillus thuringiensis Spectrum and Control Efficacy Using Cry Toxin Mixture and Immunosuppressant (Cry 독소단백질 혼합과 면역억제제 첨가를 통한 Bacillus thuringiensis 살충제 적용범위 및 방제력 증진 기술)

  • Eom, Seonghyeon;Park, Youngjin;Kim, Yonggyun
    • The Korean Journal of Pesticide Science
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    • v.18 no.3
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    • pp.181-190
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    • 2014
  • An entomopathogenic bacterium, Bacillus thuringiensis (Bt), can sporulate along with production of insecticidal Cry toxins. Bt Cry toxins exhibit relatively narrow spectrum to target insects due to their specific interactions with midgut receptors. This study designed several strategies to enhance Bt efficacy in target insect spectrum and insecticidal activity. Four Cry toxins were purified from four different Bt strains and showed relatively narrow target insect spectrum. However, the Cry mixtures significantly expanded their target insect spectra. The additional effect of baculovirus to Cry toxin was tested with recombinant baculoviruses expressing Cry1Ac or Cry1Ca. However, the baculovirus was little effective to expand target insect spectrum. Bacterial culture broth of Xenorhabdus nematophila (Xn) significantly suppressed insect cellular immune response and increased Cry toxicity. The addition of Xn culture broth to Cry mixture significantly enhanced Bt efficacy in target insect spectrum and insecticidal activity.