• Title/Summary/Keyword: EDTA-Tris

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The Effect of Antibiotics in Combination with EDTA-Tris on the Methicillin-Resistant Major Pathogens of Bovine Mastitis in Milk (유즙내에서 메티실린 내성을 지닌 젖소 유방염 주요 원인균에 대한 항생제와 EDTA-Tris의 병합의 효과)

  • Yoo, Jong-Hyun;Park, Hee-Myung
    • Journal of Veterinary Clinics
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    • v.25 no.5
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    • pp.346-354
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    • 2008
  • The combined effects of EDTA-Tris and eighteen antimicrobial agents have been evaluated in eight clinically isolated methicillin-resistant bacteria (Staphylococcus aureus, Escherichia coli, Streptococcus uberis and Streptococcus agalactiae) from bovine mastitis. The antimicrobial activity was evaluated by measuring the minimal bactericidal concentration (MBC) for the antibiotics alone or in combination with EDTA-Tris in Mueller-Hilton broth and milk. Combined use of EDTA-Tris and antibiotics potentiated or antagonized activity of antibiotics against mastitic pathogens. Milk increased the antibiotic potency of erythromycin and spiramycin on S. aureus. Culture in milk changed patterns of EDTA-Tris combinational effects compared with that in standard Mueller-Hilton broth. Combined with EDTA-Tris in milk, synergic effects were observed in colistin, dihydrostreptomycin, kanamycin, erythromycin, gentamycin, oxytetracycline, streptomycin to E. coli, Str. uberis, and Str. agalactiae. However, significant antagonistic effects of milk on antibiotic susceptibility in combination with EDTA-Tris were noted in neomycin, streptomycin, penicillin, roxithromycin, and amoxicillin. This study indicates that combination therapy of EDTA-Tris with antibiotics in bovine mastitis should be used with caution because of the possible antagonistic effects of antibiotic combination with EDTA-Tris on mastitic pathogens. In addition, antibiotic susceptibility test in combination with EDTA-Tris in milk culture condition can be benefit in search of effective treatment regimen for some antibiotic-resistant bacteria of mastitis.

Combination Effects of EDTA-Tris and Antibiotics on Bovine Mastitis Pathogens in Bovine Milk (우유즙중에서 유방염 세균에 대한 EDTA- Tris와 항생제병용의 항균효과)

  • Choi Jun-Pyo;Han Hong-Ryul
    • Journal of Veterinary Clinics
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    • v.5 no.2
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    • pp.73-81
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    • 1988
  • Combinations of EDTA-Tris and gentamicin, oxytetracycline in normal bovine milk were examined for synergistic activities aganist Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium Pyogenes, Escherichia coli, Salmonella dublin, Pseudomonas aeruginosa, and proteus spp. isolated from the milk of acute clinical bovine mastitis. The results were summarized as follows: 1. The minimal inhibitory concentrations of EDTA-Tris and gentamicin, oxytetracycline on Escherichia coli, Salmonella dublin, proteus spp., Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus agalactiae were markedly reduced. 2. The significant synergistic effects observed when the microorganisms were reacted with EDTA-Tris and gentamicin, oxytetracycline. These findings were respectively verified by kinetic studies of microbial death, using one-fourth minimal inhibitory concentrations of EDTA-Tris, gentamicin, and oxytetracycline.

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A Study on Development of Boar Semen Extender Kp for Swine AI I. Stabilization of pH Change and In Vitro Survival of Frozen-Thawed Boar Sperm in Kp Extender (돼지인공수정용 정액액상보존제 Kp 의 개발에 관한 연구 I. Kp 의 pH 조절과 냉동정자에 의한 보존성 검정)

  • 김선의;정구민;서동삼;김득중;김인철;김현종;신영수;임경순
    • Korean Journal of Animal Reproduction
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    • v.22 no.4
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    • pp.405-410
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    • 1998
  • Boar semen extender Kp (Hankook Life-Science, Korea) was newly formulated by authors. This study was carried out to investigate the optimal concentrations of EDTA, Tris and citrate buffers in the Kp extender (Basic Kp) on the pH change during storage. And then the motility of boar sperm with the Kp pH stabilized (Modified Kp) was compared with those of commercial products imported into Korea such as BTS (Mini-tube. Germany; BTSg), BTS (Tri-bio, USA; BTSa) and Modena (SGI, USA). The pH values of all extenders were increased gradually with the storage days. Especially, the initial pH of Basic Kp was higher than that of BTSg, BTSa and Modena, and also higher than physiological pH of boar sperm (6.8∼7.5). When Basic Kp was added with various concentrations of EDTA (0, 0.63, 1.25 & 2.37g/L), Tris (0, 0.18, 0.35, 0.71 & 1.42g/L) and Citrate (0, 0.75, 0.81, 1.00, 1.25 & 1.50g/L) buffers for pH down-regulation and stabilization of pH, the group added with 1.25g EDTA, 1.42g Tris and 1.00g Citrate well maintained the neutral range of pH during storage (6.88 at day1 to 7.33 at day6 in Modified Kp), Especially, the concentrations of the buffers added in Modified Kp were lower, until 1/2∼1/4 ranges, than those in Modena and other extenders. The motility of frozen-thawed boar sperm diluted with Modified Kp was significantly higher than that of Basic Kp, BTSg, BTSa and Modena (87.0% vs. 55.0∼71.0% at day1; 13.3% vs. 0∼6.3% at day6), Conclusively, Modified Kp in this experiment was kept the favorable physiological conditions in spite of low concentrations of the buffers and motility of frozen-thawed boar sperm was obtained better than that of Basic Kp and other commercial products such as BTSg, BTSa and Modena.

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Successful EDTA-Tris treatment of Pseudomonas aeruginosa infection of urinary bladder secondary to urolithiasis in a dog

  • Lee, Sang-Gwan;Hoh, Woo-Pil;Eom, Ki-Dong;Lee, Keun-Woo;Oh, Tae-Ho
    • Korean Journal of Veterinary Research
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    • v.46 no.1
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    • pp.83-86
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    • 2006
  • About 8 year-old castrated male Yorkshire terrier was presented for evaluation of dysuria, stranguria, hemtauria, and pollakiuria. On history taking, dysuria first was observed three months ago and these signs were waxed and waned. Physical examination revealed mild left perineal swelling. On routine laboratory examination, no significant findings were identified. Positive contrast urogram identified peritoneal herniation of urinary bladder. Urinalysis showed proteinuria and hematuria. Urine sediment revealed epithelial cells, white blood cells and rod-shaped bacteria. Pseudomonas aeroginosa was isolated from urine obtained through cystocentesis, and had resistance against fourteen antibiotics. Cystitis caused by P. aeruginosa concurrent with cystolithiasis and perineal hernia was diagnosed. Cystotomy, herniorrhaphy and EDTA-Tris solution lavage of bladder were performed. The patient was recovered to normal condition 2 days after treatment. Two weeks later, bacterial culture of urine was negative and any abnormality in ultrasonogram and urinalysis was not observed except calcium oxalate dihydrate crystals.

The Disruption Yeast Cell Wall by chemical Treatment (화학적 처리방법에 의한 효모의 세포벽 제거)

  • 문정혜;김중균
    • Journal of Life Science
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    • v.8 no.2
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    • pp.197-202
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    • 1998
  • The cell of Kluyveromyces fragilis yeast, which is worthy of an algal substitute, was disrupted by a chemical treatment to increase the digestion of filter-feeders that yeasts are fed to. The optimum conditions of the chemical treatment were obtained by incubating yeasts at 3$0^{\circ}C$ for one hour after treated by 1 M of Na$_{2}$-EDTA that was dissolved in 0.2 M of Tris-buffer and by 0.3 m of 2-mercaptoethanol. The percentage of protop[last production was about 30%. The percentage could be doubled by the pretreatment of three times of 30 seconds sonication.

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The effect of antioxidants on the properties of regenerated cellulose (재생셀룰로오스 제조에 미치는 산화방지제의 영향에 관한 연구)

  • Lee, Soo;Lee, Sang-Won
    • Journal of the Korean Applied Science and Technology
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    • v.27 no.3
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    • pp.378-384
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    • 2010
  • Regenerated cellulose was prepared from Buckeye wood pulp V60 via dissolution in N-methylmorpholin N-oxide (NMMO) solvent system. The effect of antioxidants such as, n-propylgallate (PG), tris(nonylphenyl) phosphite (TRIS), ethylenediamine tetraacetic acid disodium salt (EDTA), and magnesium sulfate on the properties of regenerated cellulose was studied using X-ray diffraction, copper index calculation, and viscometry. Only addition of more than 0.01% of PG into NMMO solvent was effective to avoid the reduction of the degree of polymerization(DP) of regenerated cellulose during dissolution at $110^{\circ}C$. However, the early stage(within 0.5h of dissolution process) degradation of cellulose was not prevented eventhough up to 0.5% PG was appled to hot NMMO system. In addition, to recover the expensive NMMO after cellulose regenerating process, the washing filtrate was studied using simple techniques, such as refractive index, pH, and conductivity measurements. Through conductivity measurement result, 4-time of washing was enough to remove the NMMO completely from regenerated cellulose.

Comparison of Optimal Storage Temperature and Collection Reagents for Living Bacterial Cells in Swab Samples (면봉시료에서 세균의 보존을 위한 최적 보관 온도와 채취 시약의 비교)

  • Lee, Yeong Ju;You, Hee Sang;Lee, Song Hee;Lee, So Lip;Lee, Han;Sung, Ho Joong;Kang, Hee Gyoo;Hyun, Sung Hee
    • Korean Journal of Clinical Laboratory Science
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    • v.53 no.4
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    • pp.326-332
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    • 2021
  • Swabs are useful and common sampling tools in various research fields, such as medicine, ecology, biotechnology, forensic medicine, and pollutant monitoring systems. Collection reagents are one of the essential components in sampling. It is important to develop a sample collection kit and designate an appropriate storage temperature because samples need to be stored for a long time. The purpose of this study was to identify the effects of three collection reagents and three storage temperatures on the recovery of living bacteria without media. We selected Escherichia coli and Staphylococcus aureus as representative environmental bacteria. Distilled water (DW), phosphate buffered saline (PBS), and Tris-EDTA (TE) buffer were used as collection reagents and stored at 22℃, 4℃, and -70℃ after sampling. The results of using each collection reagent and storage temperature on the bacteria were compared using relative light units (RLU) and the number of colony forming units (CFU). When using -70℃ storage temperature and the TE buffer, the number of living bacteria and the RLU values remained constant. It is therefore recommended that the sample be stored at -70℃ immediately after collection and a TE buffer solution be used as the collection reagent.

Optimization of RNA Purification Method from Ecklonia cava Kjellman (Laminariales, Phaeophyceae)

  • Ahn, Jong-Sung;Woo, Seon-Ock;Kim, Jeong-Ha;Oh, Yoon-Sik;Oak, Jung-Hyun;Yum, Seung-Shic
    • ALGAE
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    • v.19 no.2
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    • pp.123-127
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    • 2004
  • A more rapid and efficient method to extract RNA from Ecklonia cava Kjellman (Laminariales, Phaeophyceae) was introduced in this study. Each step of the procedure was evaluated and the optimal concentration of each chemical in the lysis solution was determined. Tissue pulverization with PVPP and β-mercaptoethanol in the lysis solution were not essential for RNA extraction of this species. The highest yield and purity of E. cava RNA were obtained by the lysis solution containing 1% CTAB, 1 M NaCl, 0.7% PVP, 10mM EDTA and 100mM Tris-Cl (pH 9.0). Approximately 8μg of RNA was obtained from 200 mg of ground tissue. The ratios of the absorbance at 260 nm and 280 nm were from 1.6 to 1.8 and those of at 230 nm and 260 nm were from 1.8 to 2.0. The extracted RNAs obtained in this study turned out to have a sufficient quality for cDNA synthesis.

An Alternative Method for Extracting Plasmodium DNA from EDTA Whole Blood for Malaria Diagnosis

  • Seesui, Krongkaew;Imtawil, Kanokwan;Chanetmahun, Phimphakon;Laummaunwai, Porntip;Boonmars, Thidarut
    • Parasites, Hosts and Diseases
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    • v.56 no.1
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    • pp.25-32
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    • 2018
  • Molecular techniques have been introduced for malaria diagnosis because they offer greater sensitivity and specificity than microscopic examinations. Therefore, DNA isolation methods have been developed for easy preparation and cost effectiveness. The present study described a simple protocol for Plasmodium DNA isolation from EDTA-whole blood. This study demonstrated that after heating infected blood samples with Tris-EDTA buffer and proteinase K solution, without isolation and purification steps, the supernatant can be used as a DNA template for amplification by PCR. The sensitivity of the extracted DNA of Plasmodium falciparum and Plasmodium vivax was separately analyzed by both PCR and semi-nested PCR (Sn-PCR). The results revealed that for PCR the limit of detection was $40parasites/{\mu}l$ for P. falciparum and $35.2parasites/{\mu}l$ for P. vivax, whereas for Sn-PCR the limit of detection was $1.6parasites/{\mu}l$ for P. falciparum and $1.4parasites/{\mu}l$ for P. vivax. This new method was then verified by DNA extraction of whole blood from 11 asymptomatic Myanmar migrant workers and analyzed by Sn-PCR. The results revealed that DNA can be extracted from all samples, and there were 2 positive samples for Plasmodium (P. falciparum and P. vivax). Therefore, the protocol can be an alternative method for DNA extraction in laboratories with limited resources and a lack of trained technicians for malaria diagnosis. In addition, this protocol can be applied for subclinical cases, and this will be helpful for epidemiology and control.

An Efficient Method for Co-purification of Eggshell Matrix Proteins OC-17, OC-116, and OCX-36

  • Zhang, Maojie;Wang, Ning;Xu, Qi;Harlina, Putri Widyanti;Ma, Meihu
    • Food Science of Animal Resources
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    • v.36 no.6
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    • pp.769-778
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    • 2016
  • In this study, we improved the eggshell-membrane separation process by separating the shell and membrane with EDTA solution, evaluating effects of three different extraction solutions (acetic acid, EDTA, and phosphate solution), and co-purifying multiple eggshell proteins with two successive ion-exchange chromatography procedures (CM Sepharose Fast Flow and DEAE Sepharose Fast Flow). The recovery and residual rates of eggshell and membrane separated by the modified method with added EDTA solution were 93.88%, 91.15% and 1.01%, 2.87%, respectively. Ovocleidin-116 (OC-116) and ovocalyxin-36 (OCX-36) were obtained by loading 50 mM Na-Hepes, pH 7.5, 2 mM DTT and 350 mM NaCl buffer onto the DEAE-FF column at a flow rate of 1 mL/min, ovocleidin-17 (OC-17) was obtained by loading 100 mM NaCl, 50 mM Tris, pH 8.0 on the CM-FF column at a flow rate of 0.5 mL/min. The purities of OCX-36, OC-17 and OC-116 were 96.82%, 80.15% and 73.22%, and the recovery rates were 55.27%, 53.38% and 36.34%, respectively. Antibacterial activity test suggested that phosphate solution extract exhibited significantly higher activity against the tested bacterial strains than the acetic acid or EDTA extract, probably due to more types of proteins in the extract. These results demonstrate that this separation method is feasible and efficient.