• Korean Journal of Veterinary Research
• /
• v.36 no.3
• /
• pp.681-692
• /
• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.

• Korean Journal of Food Science and Technology
• /
• v.31 no.2
• /
• pp.465-474
• /
• 1999

RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

• Korean Journal of Food Science and Technology
• /
• v.11 no.4
• /
• pp.242-248
• /
• 1979

To detect proper lytic assay conditions of the crude zumolyase from Arthrobacter luteus, effets of the various factors involved in the lytic system of Sacchromyces sake cultured with shaking in the malt extracts medium were investigated. The results are summarized as follows : 1. The susceptibilities of viable cells of S. sake from logarithmic growth phase to the lytic enzmye were much greater than those of the cells in lag and stationary phases. The cells cultured for 18 hr were the most susceptible to the enzyme. 2. Lytic activity of the enzyme toward the viable cells of S. sake was very low. It was, however, enhanced 4 folds of more by the pretreatment of the cells with 0.05 M sodium sulfite. 3. Lytic activity of the enzyme toward commercial baker's yeast cells was negligible, and the effect of sodium sulfite on the lysis of the cells also was nothing but a little. 4. The lyophilized cells of the baker's yeast showed more susceptibility to the lytic enzyme than viable cells of the yeast. No definite effect of sodium sulfite on the lysis of the lyophilized cells, however, was observed either baker's yeast of S. sake. 5. It appeared that the relationship between the reaction rate and the enzyme concentration on the lysis of the yeast cell walls followed enzyme kinetic theory, but one between the reaction rate and concentration of the yeast cells as substrates showed different pattern from that in enzyme kinetic theory. 6. After the preparation of crude zymolyase was kept at $7^[\circ}C$ for 10 days in the 0.05 M phosphate buffer, pH 7.5, the remainning lytic activity was about 80 %.

• Resources Recycling
• /
• v.29 no.2
• /
• pp.55-61
• /
• 2020

Selective catalytic reduction (SCR) has been a promising technology to reduce the air pollution caused by nitrogen oxides (NOx) in several industries. The consumption of SCR catalysts increases every year as technology evolves, however those have a limited lifespan and usually end up in landfills after they deactivate. Currently, the most widely used catalyst for and stationary applications is V₂O₅-WO₃/TiO₂ which can contain around 50% wt V₂O₅ and 7-10% wt of WO₃. The vast uses for both vanadium and tungsten and the worldwide interest in recycling methods that allow for the extraction of metals from secondary sources represent the major motivation for this research. The extraction time, pH dependency, extraction concentration studies were carried out using Aliquat 336 in exxol D80 as the extractant. It was determined that to optimize the extraction of both metals 30min of contact time with an organic phase containing 0.5mol/L of Aliquat 336 are needed at a slightly acidic pH (~5.0). In addition, counter McCabe-Thiele studies allowed us to determine that one stage is necessary for the removal of 99% of vanadium while 2 stages are necessary for the extraction of tungsten and counter current simulations proved that the theoretical approach was correct.