• Title/Summary/Keyword: Broth

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Evaluation and Optimization of a Serum-based Minimum Inhibitory Concentration Assay to Caspofungin in Candida albicans Clinical Isolates

  • Yoo, Young Bin;Kim, Sung-Soon;Kim, Young Kwon;Kim, Sunghyun
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.174-183
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    • 2016
  • In the present study, a serum-based minimum inhibitory concentration (MIC) testing to caspofungin was optimized and evaluated to solve the limitations of the conventional Clinical and Laboratory Standards Institute (CLSI) guideline-based antifungal agent MIC test and the usefulness of this testing for clinical application was determined. A total of 105 Candida albicans clinical isolates were used for measuring MIC to caspofungin. Results showed that growth characteristics were different according to types of serum and the mouse serum was the most suitable for this assay. In order to measure the optimal concentration of mouse serum, 0 to 100% mouse serum were added to the media during fungal culture. The optimal concentration of serum was 50% when consideration of antifungal agent administration and inoculum size, serum components and ease of hyphae separated, and the consideration of the degree of growth. In comparison of the usefulness between the conventional Alamar-modified broth microdilution MIC assay and 50% mouse serum-based MIC testing, the range of $MIC_{80}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.42{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $2.0{\sim}32.0{\mu}g/mL$ (SD ${\pm}9.01{\mu}g/mL$). The range of $MIC_{50}$ of the Alamar-modified broth microdilution MIC assay was $0.13{\sim}2.0{\mu}g/mL$ (SD ${\pm}0.40{\mu}g/mL$) and that of the 50% mouse serum-based MIC assay was $1.0{\sim}16.0{\mu}g/mL$ (SD ${\pm}2.36{\mu}g/mL$). The MICs of 50% mouse serum-based MIC testing was increased by up to 4 to 64 times than Alamar-modified broth microdilution MIC assay. In conclusion, a 50% mouse serum-based MIC assay was more useful for measuring MIC in Candida albicans clinical isolates than conventional colorimetric broth microdilution MIC testing.

Lactic Acid Fermentation and Biological Activities of Rubus coreanus (복분자의 유산발효와 생리활성 평가)

  • Chang, Hak-Gil;Park, Young-Seo
    • Applied Biological Chemistry
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    • v.46 no.4
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    • pp.367-375
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    • 2003
  • The puree of Rubus coreanus was fermented using lactic acid bacteria and its biological activities were examined. Lactobacillus acidophilus KCCM 32820, L. casei KCCM 12452, Lactococcus lactis subsp. lactis KCCM 40104, and Streptococcus thermophilus KCCM 40430 were used as a single or mixed starter for the lactic acid fermentation, and their cultures at the late logarithmic growth phase were inoculated to final concentration of 2% (v/v). L. casei fermented the puree of Rubus coreanus best when used as a single starter, and the culture of L. casei and L. lactis with the inoculation ratio of one to one showed the highest fermentation activity when used as a mixed starter. However, the fermented broth of the puree of Rubus coreanus using L. acidophilus and S. thermophilus showed the best results in the sensory evaluation. The optimal lactic acid fermentation conditions were as follows; the concentration of oligosaccharide added was 1% (w/v), pH of puree and fermentation temperature were 4.0 and $37^{\circ}C$, respectively, and fermentation time was $72{\sim}96$ hours. Glucose and fructose were major free sugars, and the content of lactic acid was 698.2 mg/100 g in the fermented broth. The fermented broth of the puree of Rubus coreanus showed the electron donating ability and nitrite scavenging ability with the value of 69% and 38.3% at pH 1.2, respectively. SOD-like activity and inhibitory activity on xanthine oxidase were also found in the fermented broth with the value of 60.3% and 41.8%, respectively. When the antimicrobial activities of the fermented broth were examined, it showed the highest growth inhibitory activity against Escherichia coli O-157:H7, and also contained antimicrobial activities against Salmonella typhimurium, Bacillus cereus, and Staphylococcus aureus.

Evaluation of Selective Media for Isolation of Foodborne Bacteria (주요 식중독균 분리용 선택배지의 타당성 연구)

  • Jo, Seo-Hee;Ha, Ji-Hyoung;Kim, Keun-Sung;Shim, Young-Hwan;Kwon, Ki-Sung;Han, Jeong-A;Hwang, In-Gyun;Ha, Sang-Do;Oh, Deog-Hwan
    • Journal of Food Hygiene and Safety
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    • v.22 no.4
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    • pp.388-394
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    • 2007
  • This study was conducted to evaluate the selective media listed in currently available Food Code in Korea. The 29 different types of media of five different types of foodborne bacteria including Salmonella spp., Bacillus cereus, Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus were tested in the broth and food. The recovery test for five different types of foodborne bacteria was performed in the artificially inoculated into chicken, rice, pork and mackerel. There was no significant differences in isolation capabilities among twenty nine different types of isolation selective media for five different types of foodborne bacteria in broth condition, while there was significantly a little differences in isolation capabilities among those on foods (P<0.05). The higher number of foodborne pathogens were isolated from conventional selective media approved in Food Code than newly developed selective media such as chromogenic media. This results suggest that there was differences of selectivities among currently available isolation selective media in many countries and further studies are needed to be approved by Korean Food and Drug Administration.

Ethanol Fermentation of Fusant between Heterologous Transformant of Saccharomyces cerevisiae and Candida tropicalis in Pilot Scale (Pilot Scale에서의 Fusant의 Ethanol 발효)

  • Seu, Jung-Hwn;Kim, Young-Ho;Lee, Soon-Mo;Bang, Byung-Ho
    • Microbiology and Biotechnology Letters
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    • v.17 no.1
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    • pp.14-18
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    • 1989
  • As the final experiment to assess the possibility of industrial application of FSC14-75, ethanol productivity from liquefied sweet potato starch was examined in a pilot scale of 300 liters. FSC14-75 produced 6.6%(v/v) of ethanol from 13.3% of liquefied sweet potato starch in 8 days, and the residual sugar was 3.15%. The corresponding efficiency was 70% of the theoretical maximum. Since we could isolate unicellular cell and flocculent cell from the fermentation broth, we designated them FSC14-75(S) and FSC14-75(F), respectively. We investigated ethanol productivity of FSC14-75(F) compared with that of FSC14-75(S) from liquefied potato starch in a mini·tar tormentor scale of 2.5 liters. FSC14-75(F) was found more favorable than the counterpart in terms of ethanol productivity, and produced 8.1%(v/v) of ethanol from 15% of liquefied potato starch with an efficiency of 75%. In a pilot scale fermentation with 15% of liquefied sweet potato starch, ethanol productivity of FSC14-75(F) reached maximum level of 7.7%(v/v) after 8 days, and the residual sugar was 1.9%. However, the ethanol productivity was not enhanced by a supplementary addition of Thermamyl to the fermentation broth after sterilization.

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Isolation and Charaterization of Myxobacteria with Proteolytic Activity (Myxobacteria의 Proteolytic Activity 특성)

  • Kim, Jae-Young;Chung, Jin-Woo;Cho, Kyung-Yun;Yi, Yong-Sub
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.183-188
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    • 2009
  • Seven isolates showing strong proteolytic activity, KYC 1028, 1100, 1134, 1139, 1151, 1159, and 1182, were collected. Out of them, the broth of KYC 1134 and KYC 1139 showed the high proteolytic activity measured by azocazein. To determine 16S rDNA sequences for identification, 16S rDNA of seven isolates were amplified and compared with the 16S rDNA sequences of other myxobacteria at NCBI. It is evident from the phylo-genetic tree that the isolates belong to the genus Myxococcus. Sharing high percentage similarity values with myxobacteria, the 16S rDNA sequences were involved in two species, Myxococcus macrospores and M. Fulvus. Biochemical characteristics of KYC 1134 broth, which showed the highest proteolytic activity, showed increased activity 8 times to seven days after culture, and protein production were increased gradually and stopped at five days. The broth had optimal temperature at $60^{\circ}C$ for proteolytic activity, and stability of pH was ranged from pH 5 to 10, at $50^{\circ}C$ and 60, respectively. To classify proteases being in the broth, ten inhibitors were determined and only bestatin showed 27% inhibition effect. The inhibition result demonstrates that the broth contains kinds of amino peptidases and other exopeptidases.

Identification and Fermentation Characteristics of Lactic Acid Bacteria Isolated from the Fermentation Broth of Korean Traditional Liquor, Andong-Soju (안동소주 발효액으로부터 분리한 젖산 세균의 동정 및 발효 특성)

  • Bae, Kyung-Hwa;Shin, Kee-Sun;Ryu, Hee-Young;Kwon, Chong-Suk;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.310-315
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    • 2007
  • To investigate the effect of lactic acid bacteria in Andong-Soju fermentation and traditional nuruk maturation, several lactic acid bacteria were isolated from the Andong-Soju fermentation broth and traditionally matured nuruks using Lactobacilli MRS agar containing bromocresol purple. Among the isolated bacteria, ADS-L1 showed the highest lactic acid production and was dominant species in fermentation broth. Based on physiological characteristics and 16S rDNA sequencing results, the ADS-L1 was identified as Pediococcus acidilactici. The ADS-L1 grew well at $50^{\circ}C$, and under the acidic conditions at pH 4, whereas the ADS-L1 failed to grew by treatments of 12% (w/v) ethanol or 0.01N HCl. Considering the high temperature of nuruk above $50^{\circ}C$ during nuruk maturation and the high ethanol concentration of broth above 12% at the end-stage of Andong-Soju fermentation, these results suggested that the ADS-L1 is popular in matured nuruks and plays role in the early-stage of fermentation. Analysis of pH, brix, reducing sugar content, lactic acid production, and cell growth during the cultivation of ADS-L1 further suggested that the ADS-L1 may contribute the prevention of contamination by rapid and steady acidification of broth, and do not cause problems by rapid death at the end-stage of fermentation.

Control of Ragweed (Ambrosia artemisiifolia) of Mutant N-29 Broth Filtrate of Streptomyces scopuliridis KR-001 (토양 방선균 N-29 배양 여액의 생태계교란 식물 돼지풀 방제효과)

  • Kim, Jae-Deok;Kim, Young-Sook;Kwak, Hwa-Sook;Kim, Hye-Jin;Lee, Youn-Me;Ko, Young-Kwan;Park, Kee-Woong;Choi, Jung-Sup
    • Weed & Turfgrass Science
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    • v.7 no.3
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    • pp.219-229
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    • 2018
  • We isolated Streptomyces scopuliridis KR-001 that produced herbicidal active metabolite(s) against several grass and broad leaf weeds. In this study, potential as natural herbicide of mutant N-29 broth filtrate of S. scopuliridis KR-001 was investigated to Ambrosia artemisiifilia in a greenhouse and field condition. The broth filtrate of mutant N-29 by foliar application showed a strong herbicidal activity to A. artemisiifilia with leaf stage in a greenhouse condition. Also, field trial of foliar application within treatment range had effectively controlled with early and middle stage of A. artemisiifilia at the natural habitats. Phytotoxic symptoms of mutant N-29 broth filtrate by foliar application were wilting and discoloration, and burn-down of leaves and finally plant death. These results suggest that mutant N-29 broth filtrate is considered possible as a natural herbicide for controlling environmentally friend to invasive alien plant such as A. artemisiifilia and may provide a new lead molecule for a more efficient herbicide.

Control efficacy of BtPlus against two mosquitoes, Aedes koreicus and Culex vagans (한국숲모기와 줄다리집모기에 대한 비티플러스 방제 효과)

  • Kim, Yonggyun;Minoo, Sajjadian;Ahmed, Shabbir
    • Korean journal of applied entomology
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    • v.59 no.1
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    • pp.41-54
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    • 2020
  • Two mosquito species were collected in still-water near farming area in Andong, Korea. Based on morphological characters, these two mosquitoes were identified as Aedes koreicus and Culex vagans, respectively. DNA barcode analyses supported the identification. An entomopathogenic bacterium, Bacillus thuringiensis subsp. israelensis (BtI), exhibited insecticidal activities against the two mosquito species and its virulence was more potent than that of B. thuringiensis subsp. kurstaki. It has been known that the bacterial metabolites of Xenorhabdus spp. suppress insect immunity and enhance pathogenicity of B. thuringiensis. This study tested the effect of the bacterial culture broth of Xenorhabdus spp. on enhancing BtI pathogenicity. Among three Xenorhabdus spp., culture broth of X. ehlersii (Xe) was relatively effective to enhance BtI pathogenicity against both mosquito species. Indeed, organic extracts of Xe culture broth suppressed the hemocyte-spreading behavior, suggesting the presence of immunosuppressant in the culture broth. These results suggest a formulation of BtPlus by mixing BtI spore and Xe culture broth to be applied to control the two mosquito species.

Physiological Characteristics and GABA Production of Lactobacillus plantarum K74 isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum K74의 GABA 생산 및 생리적 특성)

  • Park, Sun-Young;Shim, Hye-Young;Kim, Kee-Sung;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.31 no.2
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    • pp.143-152
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    • 2013
  • Aminobutyric acid (GABA), a major inhibitory neurotransmitter in the central nervous system of animals, has several physiological effects including anti-hypertensive, diuretic, tranquilizing, and anti-stress properties, in humans. The purpose of this study was to investigate Lactobacillus plantarum K74, which was isolated from kimchi and selected as a strain with a high ability to produce GABA, to develop a new starter culture for fermented milk production. L. plantarum K74 produced $134.52{\mu}g/mL$ GABA in MRS broth containing 1% MSG, $212.27{\mu}g/mL$ GABA in MRS broth containing 2% MSG, and $234.63{\mu}g/mL$ GABA in MRS broth containing 3% MSG. The optimum growth temperature of L. plantarum K74 was $34^{\circ}C$, reaching a pH of 4.4 after 18 hours of growth. L. plantarum K74 was most sensitive to novobiocin out of 16 different antibiotics tested, and was most resistant to kanamycin and polymyxin B. L. plantarum K74 did not produce ${\beta}$-glucuronidase, a carcinogenic enzyme, and was comparatively tolerant to bile juice and low pH. Furthermore, it displayed resistance to Escherichia coli, Salmonella typhimurium, and Staphylococcus aureus at rates of 54.9%, 46.3%, and 0.7%, respectively.

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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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