• Title/Summary/Keyword: Broth media

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Viability of eggs, filariform larvae and adults of Stronglyloides venezuelensis (Nematoda: Strongyloidea) maintained in vitro (베네수엘라분선충 (Strongvloides venezuelensis)의 충란, 감염자충 및 성충의 실험관 내 배양)

  • ;M.
    • Parasites, Hosts and Diseases
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    • v.36 no.2
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    • pp.99-108
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    • 1998
  • The present study was performed to check the viability of eggs, filariform larvae and adults of Strongvloines venezueLensis exposed to various conditions for an in vitro maintenance. The eggs in the feces remained viable for about 25 days at $4^{\circ}C$ and 15 days at room temperature. However, the isolated eggs in sterile saline lost their viability within 24 hr at $4^{\circ}C$. The eggs in morula stage were very sensitive to air drying and rapidly lost their viability (=12 hrs. Filariform larvae survived for a maximum period of 45 days in fecal suspension and 28 days in 0.12% nutrient broth in polyvinyl culture bags maintained at $20^{\circ}C$. On the other hand, those isolated from nutrient broth cultures survived for a maximum period of 32 days in tap water and 22 days in sterile saline at $20^{\circ}C$. The mature adult worms obtained from experimentally infected rats survived maximally for 9 days in serum supplemented (10% rat-serum) 0.12% nutrient broth and 4 days in serum free nutrient broth at $37^{\circ}C$ while the culture media were changed at an alternate day. The adult female worms deposited fertile eggs in serum supplemented and serum free nutrient broth cultures, however, the hatched larvae (Ll) were not able to develop to the filariform stage in the culture media and found to die within 24 hr of maintenance. The present findings on an in vitro maintenance of different stages of 5. uenezueLetis may provide useful information for biological and biochemical studies with Strongyloines species. Key words: Strongvloides venezuelensis. viability in vitro maintenance, free-living filariform larvae (L3), embryonation of eggs

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INHIBITORY EFFECT OF CRUDE IgY ON ACID PRODUCTION AND ENAMEL DEMINERALIZATION BY STREPTOCOCCUS MUTANS (Streptococcus mutans의 산 생성과 법랑질 탈회에 대한 조난황항체(IgY)의 억제 효과)

  • Oh, Se-Yeong;Lee, Kwang-Hee;Kim, Dae-Eup
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.1
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    • pp.76-83
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    • 2002
  • The purpose of study was to determine the effectiveness of crude IgY to S. mutans in preventing the acid production and the demineralization of primary tooth enamel in vitro. The acid production by S. mutans in Todd Hewitt broth with and without 5% sucrose was inhibited by 2.5% crude IgY, and as the concentration of crude IgY increased from 2.5% to 17.5%, the pH drop of the media after incubation continued to decrease. There were high positive correlations between the concentration of crude IgY and the pH of media in the late incubation period. The inhibition rate of demineralization of primary tooth enamel by S. mutans was determined by measuring the surface microhardness after incubation in 5% sucrose Todd Hewitt broth for 12 hours. The inhibition rate was 32.28% in 2.5% IgY, 42.28% in 7.5% IgY, 64.06% in 12.5% IgY, and 92.79% in 17.5% IgY. There was high positive correlation between the concentration of crude IgY and the surface microhardness of enamel after demineralization These results suggest that it would be possible to prevent dental caries through passive immunization using crude IgY.

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Occurrence of Glutathione Sulphydryl (GSH) and Antioxidant Activities in Probiotic Lactobacillus spp.

  • Yoon, Yung H.;Byun, Jung R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.11
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    • pp.1582-1585
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    • 2004
  • The antioxidative ability on the basis of reduced glutathione sulphydryl level, the inhibition activities of linoleic acid peroxidation of cell free extract of Lactobacillus spp. and the effects of types of media and growth phase of the cells on the cellular GSH level have been determined. Correlation between reduced glutathione sulphydryl level and antioxidative ability of Lactobacillus spp. was analyzed: Lactobacillus casei HY 2782 contained 25.15 $\mu$mole/g of GSH, the cellular GSH level of L. casei HY 2782 reached maximum after 24 h of cultivation and tended to decrease on further cultivation up to 72 h. There was a significantly higher level of cellular GSH when grown in de Man Rogosa and Sharpe (MRS) broth than in tryptone phytone yeast extract (TPY) broth or bromcresol pruple dextrose (BCP) broth (p<0.05). The antioxidant activity of cell free extract of Lactobacillus spp. have been shown to be significantly different among strains in the inhibition of linoleic acid peroxidation by thiobarbituric acid (TBA) test (p<0.01). L. casei HY 2782 and L. acidophilus ATCC 4356 revealed a high degree of antioxidative effect in linoleic acid oxidation system. Spearmans' rank correlation coefficient between inhibitory activity on linoleic acid peroxidation and cellular GSH levels of Lactobacillus spp. was 0.65, which means a significant positive correlation.

Effect of Cellobiose Octaacetate, Avicel, and KC-flock on Production of Avicelases from Penicilliurn verruculosum (Penicillium verruculosum의 Acicelase 생성에 대한 Cellobiose Octaactate와 Avicel 및 KC-flock 의 영향)

  • 조남철;김강화;전순배;정기철
    • Microbiology and Biotechnology Letters
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    • v.18 no.4
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    • pp.383-389
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    • 1990
  • During the cultivation of Penkillium uerrmulosum in the media containing cellobiose octaacetate (COA), avicel, or KC-flock as an inducer and as a sole carbon source for 21 days, cellulolytic activity and SDS-PAGE pattern of proteins in the culture broth were investigated. Protein concentration and cellulolytic activity were highest in the COA medium. As cultivation period was increased, protein content and avicel hydrolytic activity of culture broth were increased as similar extent but neither $\beta$-glucosidase nor CMC hydrolytic activity was correlated to protein content. When crude proteins from the culture broth were separated on DEAE column by HPLC, distribution of avicel-hydrolytic activities were well correlated with that of major proteins. From those results it was suggested that three major proteins having 60 K, 68 K, and 76 K of Mr. were avicel-hydrolytic enzymes.

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Optimization of Indole-3-acetic Acid (IAA) Production by Bacillus megaterium BM5

  • Lee, Jae-Chan;Whang, Kyung-Sook
    • Korean Journal of Soil Science and Fertilizer
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    • v.49 no.5
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    • pp.461-468
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    • 2016
  • One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

Effect of Ammonium Phosphate on Mycelial Growth and Exopolysaccharides Production of Ganoderma lucidum in an Air-Lift Fermenter

  • Lee, Kyu-Min;Lee, Shin-Young;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.726-731
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    • 1999
  • It was discovered that ammonium phosphate in the medium played an important role in both growing mycelium and producing exopolysaccharides (EPS) from G. lucidum. In lower concentration levels of ammonium phosphate (0-3 g/l), an improved mycelial growth was observed by maintaining more filamentous morphology than in high concentrations (5-11 g/l). In addition, it was confirmed by comparing the factual dimension and frequency of the area regarding the mycelial pellets. This must be attributed to limitations of nutrient transfer by maintaining filamentous mycelium during the cultivation in a low ammonium phosphate containing medium. On the other hand, the best EPS production was observed in medium with the absence or low concentration of ammonium phosphate. The shear stress of the culture broth was greatly affected by the shear rate, as compared with that of the culture broth with high ammonium phosphate concentration. The rheological characteristics of the fermentation broth and filtrate worked well according to the Herschel-Bulkley model. It was also found that the morphological changes of the mycelium resulting from the ammonium phosphate concentration directly affected the rheological characteristics of the system and resulted in reversely affecting the EPS production levels. Based on these results, it can be concluded that delicate regulation of the ammonium phosphate concentration in the culture media should be provided in order to obtain optimal mycelial growth and/or EPS production.

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Antimycotic Susceptibility Testing of Trichophyton Rubrum by Microculture Method (96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사)

  • Lee, Moo-Woong;Kim, Jong-Chul;Choi, Jong-Soo;Kim, Ki-Hong
    • Journal of Yeungnam Medical Science
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    • v.9 no.2
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    • pp.396-406
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    • 1992
  • Various susceptibility tests have been used to determine minimal inhibition concentration(MIC) of dermatophytes. They have limitations to apply practically because they need long time to determine MIC. Authors examined MIC of T. rubrum to ketoconazole and itraconazole using 96-well microplate and 24-well macroplate by method of Granade and Artis and tried to check the possibility of this method on clinical application. Nine strains of T. rubrum from patients with dermatophytosis were used. Evaluations of the factors affecting MIC were also tried. The results were as follows. 1. Effect of inoculation density on determination time and MIC : Determination of MIC were possible in 4th days after inoculation at higher inoculation density Caborbance 2.0, 1.0) compared to 6th days at lower inoculation density(absorbance 0.5, 0.25). 2. Effect of incubation temperature on MIC : When incubating at $37^{\circ}C$, MIC were below 0.006-$0.04{\mu}g/ml$ to ketokckonazole and below 0.006-$0.04{\mu}g/ml$ to itraconazole while at $25^{\circ}C$ 0.08-$5.68{\mu}g/ml$ to ketoconazole and 0.006-$0.71{\mu}g/ml$ to itraconazole. Significant reduction of MIC was observed at $37^{\circ}C$ compared to $25^{\circ}C$. 3. Effect of container size on determination time and MIC : When incubating in 96-well microplate and 24-well macroplate, determination of MIC was possible in 4th to 6th days after inoculation in broth-containig 96-well microplate compared to 8th to 12th days in broth-containing 24-well macroplate. But no difference in MIC was observed between different container size. 4. Effect of media on MIC : When using broth as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconazole, below 0.006-$0.36{\mu}g/ml$ to itraconazole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconzole, below 0.006-$5.68{\mu}g/ml$ to intraconzole in agar-containing 24-well macroplate. There was slight increase of MIC with agar media compared to broth media. 5. These findings confirm that determination of MIC of dermatophtes by method of Granade and Artis is fast and simple technique for antifungal susceptibility test.

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Comparison of Real-Time PCR and Culture Methods for Detection of Campylobacter jejuni in Various Foods (다양한 식품에서 Campylobacter jejuni 검출을 위한 real-time PCR과 배지배양법의 비교검증)

  • Chon, Jung-Whan;Hyeon, Ji-Yeon;Hwang, In-Gyun;Kwak, Hyo-Sun;Han, Jeong-A;Kim, Moo-Sang;Kim, Jong-Hyun;Song, Kwang-Young;Seo, Kun-Ho
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.119-123
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    • 2011
  • In this study, performances of culture methods using two selective media and real-time PCR were evaluated for detection of Campylobacter jejuni (C. jejuni) in various food samples. Sausage, ground beef, and radish sprouts inoculated with C. jejuni were enriched in Hunt broth and then streaked onto modified cefoperazone charcoal deoxycholate agar and Preston agar, followed by incubation under microaerobic conditions. The enriched Hunt broth (1 mL) was used in real-time PCR assay. No statistical differences were observed in sensitivity among the two selective media and real-time PCR for sausage and ground beef. However, the number of positives by real-time PCR in radish sprouts was much higher than the two selective media (p<0.05). It appears that real-time PCR could be used as an effective screening tool to detect C. jejuni, particularly in foods with a high number of background microflora such as fresh vegetables.

Isolation and Cultural Characteristics of a Phosphate-solubilizing Fungus, Penicillium sp. PS-113 (인산가용화 사상균, Penicillium sp. PS-113 균주의 분리 및 배양특성)

  • 강선철;최명철
    • KSBB Journal
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    • v.13 no.5
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    • pp.497-501
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    • 1998
  • A fungus with high phosphate solubilizing activites was isolated from soil using potato dextrose agar-calcium phosphate medium and identified to Penicillium sp. PS-113, based on the morphological characteristics of conidiophore and conidia; flask shape of phialide, simple branching type of conidiophore, and columnar shape of conidial head, in malt extract agar and potato dextrose agar media. The optimum temperature ad initial pH to solubilize rock phosphate in potato dextrose broth-rock phosphate medium were 30$^{\circ}C$ and pH 8.0, respectively. In these conditions phosphate solubilizing activities of Penicillium sp. PS-113 against four types of insoluble phosphate like tricalcium-phosphate, aluminium phosphate, hydroxyapatite and rock phosphate were quantitatively determined. As results, this fungus highly produced free phosphates to the culture broth with the concentrations of 1,283 ppm against tricalcium-phosphate, 585 ppm against rock phosphate, 528 ppm against aluminium phosphate, and 242 ppm against hydroxyapatite, respectively.

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Immunological ObservatioIn on Gandidiasis (Candida증(症) 발생(發生)의 면역학적(免疫學的) 기전(機轉)에 관(關)한 연구(硏究))

  • Kim, Hong-Sik
    • The Korean Journal of Mycology
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    • v.6 no.2
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    • pp.15-18
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    • 1978
  • The study was performed to investigate the inhibitory action on the growth rate of Candida albicans under the various immunological conditions, Candida albicans was grown in broth media containing 0.1ml. (in total broth volume 4.0ml.) of normal human serum, diabetic patient serum, albumin solution, artifical bulla content and nutrient solution as control subject. The inhibitory action of Candida albicans was indicated by mesurement of transparency rate with use of the spectrophotometer. The results are as follows: 1. Normal human serum shows inhibitory effect on the growth of the candida distinctly. 2. Albumin solution reveals almost similar to that of the effect of normal human serum. 3. Artificial bulla content which obtained by irradiation of ultraviolot ray after application of 1% 8-Methoxy-psolaren cream and the diabetic patient serum shows reduction of inhibitory effect as compared with that of the normal human serum. 4. It is estimated that the titer of negative effect of diabetic patient serum is not related with the variation of immunoglobulin titer in patient serum.

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