• Title/Summary/Keyword: Hemolysis reaction

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Prevalence of Listeria spp in intestinal contents of slaughtered cattle (도축우의 장내용물에서 Listeria속균의 분포도 조사)

  • Lim Hyun-Sook;Suh Dong-Kyun
    • Korean Journal of Veterinary Service
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    • v.28 no.2
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    • pp.113-119
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    • 2005
  • We surveyed the distribution of Listeria spp in intestinal contents of slaughtered cattle from Daegu between March and October 2003. Fourteen Listeria spp were isolated from a total of 100 samples. Two samples contained only L innocua and other six samples contained both L monocytogenes and L innocua. Of the 99 samples positive to esculin reaction in Fraser broth, Listeria spp were isolated only from $8\%$ of the samples. Three selective plating medium were examined for detection of Listeria species including Enhanced hemolysis agar, Oxford agar and Palcam agar, It was found that Enhanced hemolysis agar was more effective than Oxford agar and Palcam agar, and that L monocytogenes needed 48 hour growth to give positive reaction.

Delivery System of Daunorubicin by Red Blood Cells (적혈구를 이용한 Daunorubicin의 배송시스템)

  • Ham, Seong-Ho;Song, Kyung;Ko, Gun-Il;Kim, Jae-Baek;Sohn, Dong-Hwan
    • Journal of Pharmaceutical Investigation
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    • v.24 no.3
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    • pp.131-137
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    • 1994
  • Drug delivery system by the use of red blood cells was established to sustain the release of drugs in the circulatory system by the intravenous injection. The entrapment method by the preswelling technique was re-examined and evaluated for searching the new entrapping conditions without hemolysis. The addition of 4 volume of $0.6{\times}\;hank's$ balanced salt solution (HBSS) into 1 volume of 50% red blood cells suspension did not induce the hemolysis and change the hematocrit level in this experimental condition (within 15 min). Most of daunorubicin could be entrapped into red blood cells within 15 min. While the intracellular adenosine triphosphate (ATP) level followed by the entrapment was reduced to 86% of normal ATP level, the membrane fluidity and the shape factor of red blood cells were not altered. The release rate of daunorubicin from red blood cells was affected by the hemolysis under this condition. To maintain the intracellular ATP in red blood cells, the new reaction buffer was made With the addition of ATP and sodium pyruvate during the entrapment procedure because the hemolysis during the release test would reflect the loss of intracellular ATP that might result in the decrease of the viability in vivo. The addition of ATP raised the intracellular ATP level, which protect the hemolysis during the release test.

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Development of Differential Media and Multiplex PCR Assays for the Rapid Detection of Listeria monocytogenes (Listeria monocytogenes의 신속검출을 위한 선택배지 및 multiplex PCR 기법 개발)

  • Jung, Byeong-yeal;Lim, Hyun-sook;Jung, Suk-chan
    • Korean Journal of Veterinary Research
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    • v.43 no.2
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    • pp.231-237
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    • 2003
  • Listeria (L.) monocytogenes in samples could not be detected occasioally by faster growth of other Listeria spp. especially L. innocua. The aim of this study was to develop the differential media and multiplex polymerase chain reaction (PCR) assays for the rapid detection of L. monocytogenes. L. monocytogenes colonies were characterized by their ${\beta}$-hemolysis with fluorescence under 366 nm UV light on the Listeria hemolysis agar (LHA). L. innocua, a species commonly present in foods, did not produce ${\beta}$-hemolysis on LHA. Therefore, one or more colonies of L. monocytogenes were easily distinguished from large populations of L. innocua. The multiplex PCR assays were developed to distinguish from L. monocytogenes and other Listeria spp. with two pairs of primers. The primers were designed in 16S rRNA and listeriolysin O gene for specific amplification of all members of the genus Listeria and L. monocytogenes, respectively. The multiplex PCR assays produced 560 and 938 bp products in L. monocytogenes; only 938 bp products in the genus Listeria. The multiplex PCR assays could detect as little as 50 pg of L monocytogenes DNA. These results indicated that the differential media and multiplex PCR assays might be useful diagnostic tools for the rapid detection of L. monocytogenes.

Nonhemolytic entrapping method into red blood cells and its release pattern (적혈구내 비용혈 약물봉입과 약물방출)

  • Ham Seong ho;Ko Geon Il;Kim Jae Baek;Sohn Dong Hwan
    • Korean Journal of Clinical Pharmacy
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    • v.5 no.2
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    • pp.75-84
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    • 1995
  • Drug delivery by red cells was established to maintain the release of drugs in the blood. The entrapment method by amphotericin B was re-examined and evaluated for obtaining the suitable entrapping conditions without hemolysis. The amphotericin B treatment below $10{\mu}g/ml$ induced the non-hemolysis to entrap daunorubicin into red cells within 10min. Under these conditions intracellular ATP level was decreased as $18\%$. Membrane fluidity and the shape factor of red cells were maintained. To maintain intracellular ATP, ATP and sodium pyruvate were added during the entrapment procedure because hemolysis during the release test would reflect the loss of intracellular ATP that would be postulate the decrease of the viability invivo. Consequently, the addition of ATP in the reaction solution can raise the intracellular level of ATP.

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Antioxidant activity of water and alcohol extracts of Thuja orientalis leaves

  • Nizam, Iram;Mushfiq, M
    • Advances in Traditional Medicine
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    • v.7 no.1
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    • pp.65-73
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    • 2007
  • Water and alcohol extracts were prepared from dried and powdered leaves of Thuja orientalis (T. orientalis). The reducing power, total phenolic content, the 1,1-diphenyl-2-picrylhydrazyl scavenging activity, inhibitory effect on Fe (II)-EDTA-$H_{2}O_{2}$ (Fenton reaction system) induced DNA damage and inhibitory effect on human red blood cell (RBC) hemolysis were evaluated in the present study. At a concentration of 200 mg, water and alcohol extracts of T. orientalis inhibited the hydrolysis of DNA by 72.859% and 65.312%, respectively. Water and alcohol extracts of T. orientalis also inhibited 2,2'-Azobis(2-amidinopropane) dihydrochloride induced RBC hemolysis to the extent of 69.30% and 54.55%, respectively. The reducing power and antioxidative activity of water extract was found to be more than that of alcohol extract. This is attributable to the presence of higher amount of phenolic compounds in water extract. The present results indicate that the T. orientalis extracts are rich sources of natural antioxidants and can protect DNA and human red blood cells against free radical induced oxidative damage.

Studies on hemolytic reaction of normal chicken serum and sheep erythrocytes (닭혈청(血淸)에 의한 면양적혈구(緬羊赤血球)의 용혈(溶血)에 관한 연구(硏究))

  • Seo, Ik Soo
    • Korean Journal of Veterinary Research
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    • v.14 no.1
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    • pp.77-90
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    • 1974
  • Hemolytic reaction of normal fresh chicken serum on sheep erythrocytes was studied and the following experimental results were obtained and summarized. 1. Chicken sera, 258 (78%) out of 344 samples showed hemolytic activity on sheep erythrocytes. 2. Distribution of a different hemolytic titer of chicken sera was not dependent to sex and age difference of test chicken. 3. Hemolytic activity of serum component obtained from normal fresh chicken was heat inactivated at $56^{\circ}C$. 30 minutes heating. 4. The most enhanced hemolytic activity of chicken serum on sheep erythrocytes was observed at the incubation temperature of $46^{\circ}C$. 5. The most effective pH for the hemolytic reaction of chicken serum on sheep erythrocytes was observed at 7.0, and pH 6.0 or 8.5 resulted less or no hemolysis. 6. Hemolytic reaction of chicken serum and sheep erythrocytes required Mg⧻ and Ca⧻ ions as, co-factor, and the former was required more compared to the latter. 7. Hemolytic activity of chicken serum was observed in ChC 2, 4 fraction but not in ChC 1, 3, ChC 3, 4, ChC 1, 2, 4 and ChC 1, 2, 3 fractions. 8. In electron micrography, morphological changes of sheep erythrocyte membrane by normal chicken serum was similar to that of immune hemolysis: that was, the hemolytic hole was circular and it was surrounded with a white ring. 9. Electron micrography of morphological changes on sheep erythrocyte membrane indicated that the size of hemolytic hole and white ring were functional to the chicken serum concentration used and reaction time.

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Studies of Coagulase Production and Isolation of R-plasmid from Staphylococcus aureus (Staphylococcus aureus의 Coagulase 생성능과 R-플라스미드 분리에 관한 연구)

  • Yoon, Hyo-Sook;Lee, Hyung-Hoan;Kim, Soo-Young
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.259-266
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    • 1987
  • A total of 129 clinical isolates of Staphylococcus species was characterized by the tests of coagulase production, haemagglutination, mannitol fermentation, DNase production and hemolysis. Ninety-nine out of them showed positive reactions to the tests, therefore they were identified as Staphylococcus aureus. The isolates showing positive reaction in haemagglutination test also showed 100% of tube coagulase positive reaction. The haemagglutination test was a reliable method for identifying Staphylococcus aureus in the clinical laboratory. S. aureus produced stronger hemolysis with human blood agar than with sheep blood agar. Antibiotic resistant S. aureus isolates(S-46, S-112, S-126) had 4 to 6 p]asmid DNA elements. The S-112 strain had 6 plasmid DNA elements(1.8, 2.2, 3.7, $26.3{\sim}50$, and 70 Mdaltons), the S-126 had 4 elements(2.6, 4.2, $4.6{\sim}60Md$), and the S-46 had 1 element(${\sim}100Md$). PPSA strain had 4 plasmid DNA elements(2.5, 4.2, $4.6{\sim}60Md$) and S. aureurs(ATCC) strain contained 9.4, 26.3 and ${\sim}50Md$ plasmid DNA elements.

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Studies on Hemolysis of Vibrio Parahaemolyticus to Various Erythrocytes (각종 적혈구에 대한 장염비브리오의 용혈성에 관한 연구)

  • Ju, Jin-Woo;Kim, Young-Hee
    • The Journal of the Korean Society for Microbiology
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    • v.19 no.1
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    • pp.49-54
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    • 1984
  • The authors isolated Vibrio parahaemolyticus from sea water, sea mud and various marine products in Busan shore area from 1981 to 1982, Among 100 isolated strains, 66 strains showed positive reaction in Kanagawa phenomenon. With the above 66 strains, the authors carred out test for detecting hemolysis activity of V.parahaemolyticus on human, rabbit, chicken, pig, goat, sheep and cow erythrocytes, in different media, such as modified Wagatsuma, nutrient, peptone and brain heart infusion agar plates media. The following results were obtained: 1. The media which can be used for Kanagawa phenomenon of V. parahoemolyticus were modified Wagatsuma, nutrient, peptone agar media, but not brain heart infusion agar medium. 2. The erythrocytes which showed positive Kanagawa phenomenon were those of human, rabbit, chicken and pig, but sheep, goat and cow erythrocytes showed no sensitivities.

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Studies on the Simplified Hemagglutination Reaction to Pasteurella multocida and the Stabilization of Erythrocytes (Pasteurella multocida에 대(對)한 간이적혈구(簡易赤血球) 응집반응(凝集反應)과 적혈구(赤血球)의 안정화(安定化)에 관(關)한 연구(硏究))

  • Lee, Hak Cheul;Chung, Yung Gun;Kim, Kyo-Joon
    • Korean Journal of Veterinary Research
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    • v.10 no.1
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    • pp.11-23
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    • 1970
  • Recently Carter(1952) reported the capsule antigens of Pasteurella multocida could be divided into four serological types A,B,C and D by means of precipitation tests. Subsequently he showed that the most sensitive for identification of these types involved the use of capsule substance adsorbed by erythrocytes in hemagglutination test. It may be somewhat difficult to conduct the hemagglutination test in small laboratory, because relatively large amounts of antisera and erythrocytes of the human O type are required for the test. A simple method for serological typing of P. multocida was the slide agglutination test employed by Little et al. (1943) and Namioka et al. (1962), but this method is still in controversy. The author tried adapting Carter's hemagglutination method to the slide method so called "micromethod technique", and studied on the stabilization of erythrocytes for use of slide hemagglutination to P. multocida although many invesigators reported the stabilization of erythrocytes. The results obtained are summarized as follows: 1. A simplified method (slide method) for capsule typing of the organism was developed by adapting Carter's hemagglutination reaction(tube method). Antibody-containing serum can be diluted serially on Boerner's microtest slide with capillary or serological pipetts with a considerable accuracy. The slide reaction can be carried out with case on the slide by adding $0.05m{\ell}$ of antigen-sensitized erythrocytes suspension diluted to one percent on $0.05m{\ell}$ of serially diluted antibody-containing sera, and the final result can be read after 60 minutes at the room temperature ($15^{\circ}C$). 2. It is difficult to determine superiority of inferiority between the slide method and the tube method on the pattern of the reaction of hemagglutination. 3. The pH range of 6.6 to 8.3 is optimal for the slide hemagglutination reaction. 4. The antigen-sensitization against erythrocytes at $37^{\circ}C$ is optimal for the slide hemagglutination. 5. Both the doses and concentration of antigen do not influence the antigen-adsorbing capacity of erythrocytes. 6. The reduction of antigen-sensitizing hours does not influence the antigen-adsorbing capacity of erythrocytes even 30 minutes. 7. The tannic acid treatment against formalinized and non-formalinized erythrocytes showed no effect on the reaction of hemagglutination. 8. The erythrocytes preserved at $4^{\circ}C$ in the ACD solution do not decrease the reactivity on the reaction of hemagglutination for 60 days, while they begin slight hemolysis 30 days after preserving. 9. The stable preparation of erythrocytes can be obtained by treating the cells at $37^{\circ}C$ for 20 hours with from 4 to 8 percent of formalin in saline or buffer. These cells can be preserved at $4^{\circ}C$ for more than 8 months experimented without hemolysis. With low concentration of formalin, the cells were not sufficiently stabilized resulting in the hemolysis after short period of preservation at $4^{\circ}C$. 10. The erythrocytes treated with 16 percent of formalin remain constantly or increase the reactivity for the reaction of hemagglutination. On the contrary, the cells treated with I to 8 percent of formalin decrease the reactivity. 11. There is no difference between nontreated fresh erythrocytes and the erythrocytes preserved in the ACD solution on the reactivity against the hemagglutination, and the erythrocytes treated with 16 percent of formalin showed the reactivity of higher level than that of the above two kinds of erythrocytes. 12. There is no difference between the saline and the isotonic buffer solution on the reaction of hemagglutination.

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Comparison of Efficiency between Pre-storage and Post-storage filtration by Leukoreduction Blood Filter

  • Shin, Geon Sik;Kim, Bohee;Kim, Sung Hoon;Rhee, Ki-Jong;Kim, Yoon Suk
    • Biomedical Science Letters
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    • v.23 no.2
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    • pp.111-117
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    • 2017
  • Leukocytes in blood components are involved in diverse adverse transfusion reactions such as febrile non-hemolytic transfusion reactions. Therefore, leukocyte reduction is required to reduce these adverse reactions. The objective of this study was to compare the efficiency of pre-storage filtration and post-storage filtration. Filtration time, residual leukocyte count, RBC recovery, and hemolysis were assessed after pre-storage or post-storage filtration. Compared to pre-storage filtration, filtration time was prolonged and hemolysis was dramatically increased when post-storage filtration was performed. Residual leukocytes count and RBC recovery after post-storage filtration were similar with those obtained after pre-storage filtration. These results suggest that pre-storage filtration has better efficiency than post-storage filtration. These are thought to contribute to the production of better quality of leukoreduction blood components.