• Title/Summary/Keyword: broth culture

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Red fluorescence of oral bacteria is affected by blood in the growth medium (성장배지 혈액 유무가 구강미생물의 적색 형광 발현에 미치는 영향)

  • Jeong, Seung-Hwa;Yang, Yong-Hoon;Lee, Min-Ah;Kim, Se-Yeon;Kim, Ji-Soo
    • Journal of Korean Academy of Oral Health
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    • v.41 no.4
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    • pp.290-295
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    • 2017
  • Objectives: Dental plaque emits red fluorescence under a visible blue light near the ultra-violet end of the light spectrum. The fluorescence characteristics of each microorganism have been reported in several studies. The aim of this study was to evaluate changes in red fluorescence of oral microorganisms that is affected by blood in the culture media. Methods: The gram-positive Actinomyces naeslundii (AN, KCTC 5525) and Lactobacillus casei (LC, KCTC 3109) and gram negative Prevotella intermedia (PI, KCTC 3692) that are known to emit red fluorescence were used in this study. Each bacterium was activated in broth and cultivated in different agar media at $37^{\circ}C$ for 7 days. Tryptic soy agar with hemin and vitamin $K_3$ (TSA), TSA with sheep blood (TSAB), basal medium mucin (BMM) medium, and BMM with sheep blood (BMMB) were used in this study. Fluorescence due to bacterial growth was observed under 405-nm wavelength blue light using the quantitative light-induced fluorescence-digital (QLF-D) device. The red, green, and blue fluorescence values of colonies were obtained using image-analysis software and the red to green ratio (R/G value) and red to total RGB ratio (R/RGB value) were calculated for quantitative comparison. Results: The QLF-D images of the AN, LC, and PI colonies showed red fluorescence in all media, but the fluorescence of all bacteria was reduced in TSA and BMM media, compared with in TSAB and BMMB media. Both the R/G and the R/RGB values of all bacteria were significantly reduced in growth media without blood (P<0.001). Conclusions: Based on this in vitro study, it can be concluded that red fluorescence of oral bacteria can be affected by growth components, especially blood. Blood-containing medium could be a significant factor influencing red fluorescence of oral bacteria. It can be further hypothesized that bleeding in the oral cavity can increase the red fluorescence of dental plaque.

Isolation of a Pseudoalteromonas sp. JH-1 Producing Agarase and Characterization of its Agarase (Agarase를 생산하는 Pseudoalteromonas sp. JH-1의 분리·동정 및 agarase의 특성 연구)

  • Lee, Dong-Geun;Kim, Ju-Hui;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.31 no.5
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    • pp.496-501
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    • 2021
  • In this study, the marine agar-degrading bacterium Pseudoalteromonas sp. JH-1 was isolated, and its growth and agarase properties were investigated. Seawater was collected from the offshore of the Yonggung Temple in Busan, and agar-degrading bacteria were isolated and cultured with marine agar medium. The bacterium Pseudoalteromonas sp. JH-1 was isolated through 16S rRNA gene sequencing. The extracellularly secreted enzyme was obtained from the culture broth of Pseudoalteromonas sp. JH-1 and was used to characterize its agarase. The extracellular agarase exhibited a maximum activity of 116.6 U/l at 50℃ and pH 6.0 of 20 mM Tris-HCl buffer. Relative activities were 31, 59, 94, 100, 45, and 31% at 20, 30, 40, 50, 60, and 70℃, respectively. Relative activities were 49, 85, 100, 86, 81, and 67% at pH 4, 5, 6, 7, 8, and 9, respectively. Residual activity was more than 85% after exposure at 20, 30, and 40℃ for 2 hr, and more than 82% after exposure at 50℃ for 2 hr. Zymogram analysis confirmed that Pseudoalteromonas sp. JH-1 produced at least two agarases of 55 and 97 kDa. As the products of α-agarase and β-agarase have antioxidation, antitumor, skin-whitening, macrophage activation, and prebiotic effects, further studies are needed on the agarase of Pseudoalteromonas sp. JH-1.

Characterization of exopolysaccharide-producing lactic acid bacteria from Taiwanese ropy fermented milk and their application in low-fat fermented milk

  • Ng, Ker-Sin;Chang, Yu-Chun;Chen, Yen-Po;Lo, Ya-Hsuan;Wang, Sheng-Yao;Chen, Ming-Ju
    • Animal Bioscience
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    • v.35 no.2
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    • pp.281-289
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    • 2022
  • Objective: The aim of this study was to characterize the exopolysaccharides (EPS)-producing lactic acid bacteria from Taiwanese ropy fermented milk (TRFM) for developing a clean label low-fat fermented milk. Methods: Potential isolates from TRFM were selected based on the Gram staining test and observation of turbid suspension in the culture broth. Random amplified polymorphic DNA-polymerase chain reaction, 16S rRNA gene sequencing, and API CHL 50 test were used for strain identification. After evaluation of EPS concentration, target strains were introduced to low-fat milk fermentation for 24 h. Fermentation characters were checked: pH value, acidity, viable count, syneresis, and viscosity. Sensory evaluation of fermented products was carried out by 30 volunteers, while the storage test was performed for 21 days at 4℃. Results: Two EPS-producing strains (APL15 and APL16) were isolated from TRFM and identified as Lactococcus (Lc.) lactis subsp. cremoris. Their EPS concentrations in glucose and lactose media were higher than other published strains of Lc. lactis subsp. cremoris. Low-fat fermented milk separately prepared with APL15 and APL16 reached pH 4.3 and acidity 0.8% with a viable count of 9 log colony-forming units/mL. The physical properties of both products were superior to the control yogurt, showing significant improvements in syneresis and viscosity (p<0.05). Our low-fat products had appropriate sensory scores in appearance and texture according to sensory evaluation. Although decreasing viable cells of strains during the 21-day storage test, low-fat fermented milk made by APL15 exhibited stable physicochemical properties, including pH value, acidity, syneresis and sufficient viable cells throughout the storage period. Conclusion: This study demonstrated that Lc. lactis subsp. cremoris APL15 isolated from TRFM had good fermentation abilities to produce low-fat fermented milk. These data indicate that EPS-producing lactic acid bacteria have great potential to act as natural food stabilizers for low-fat fermented milk.

Occurrence of the Onion Moth, Acrolepiopsis sapporensis, in the Welsh Onion Farms and its Treatment Using 'BtPlus' (대파 재배지 파좀나방(Acrolepiopsis sapporensis) 발생 현황과 '비티플러스' 처리 효과)

  • Md Tafim Hossain Hrithik;Gahyeon Jin;Yonggyun Kim
    • Korean journal of applied entomology
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    • v.62 no.4
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    • pp.277-285
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    • 2023
  • The onion moth, Acrolepiopsis sapporensis, was monitored in the farms cultivating the welsh onion, Allium fistulosum, using sex pheromone from transplantation to harvest. Two occurrence peaks were observed at early June and late July after the overwintering population. However, the population sizes were varied among different years and the cultivating environments. To effectively control A. sapporensis with microbial pesticides, different Bacillus thuringiensis strains were screened to select B. thuringiensis kurstaki (BtK). To enhance the insecticidal virulence of BtK, the culture broth of Photorhabdus temperata temperata (Ptt) was added to the BtK. This mixture of two entomopathogenic bacteria was called 'BtPlus', which was superior to BtK alone in the insecticidal virulence. The enhanced virulence was explained by the immunosuppressive activity of the secondary metabolites contained in the Ptt extract. The metabolites inhibited both cellular and humoral immune responses of A. sapporensis, resulting in the enhanced virulence of BtK. These results suggest that A. sapporensis occurs in the welsh onion fields and the resulting economic damage would be effectively prevented by BtPlus application.

Process Optimization of Dextran Production by Leuconostoc sp. strain YSK. Isolated from Fermented Kimchi (김치로부터 분리된 Leuconostoc sp. strain YSK 균주에 의한 덱스트란 생산 조건의 최적화)

  • Hwang, Seung-Kyun;Hong, Jun-Taek;Jung, Kyung-Hwan;Chang, Byung-Chul;Hwang, Kyung-Suk;Shin, Jung-Hee; Yim, Sung-Paal;Yoo, Sun-Kyun
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1377-1383
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    • 2008
  • A bacterium producing non- or partially digestible dextran was isolated from kimchi broth by enrichment culture technique. The bacterium was identified tentatively as Leuconostoc sp. strain SKY. We established the response surface methodology (Box-Behnken design) to optimize the principle parameters such as culture pH, temperature, and yeast extract concentration for maximizing production of dextran. The ranges of parameters were determined based on prior screening works done at our laboratory and accordingly chosen as 5.5, 6.5, and 7.5 for pH, 25, 30, and $35^{\circ}C$ for temperature, and 1, 5, and 9 g/l yeast extract. Initial concentration of sucrose was 100 g/l. The mineral medium consisted of 3.0 g $KH_2PO_4$, 0.01 g $FeSO_4{\cdot}H_2O$, 0.01 g $MnSO_4{\cdot}4H_2O$, 0.2 g $MgSO_4{\cdot}7H_2O$, 0.01 g NaCl, and 0.05 g $CaCO_3$ per 1 liter deionized water. The optimum values of pH and temperature, and yeast extract concentration were obtained at pH (around 7.0), temperature (27 to $28^{\circ}C$), and yeast extract (6 to 7 g/l). The best dextran yield was 60% (dextran/g sucrose). The best dextran productivity was 0.8 g/h-l.

Development of Optimal Culture Media for the Stable Production of Mushroom (버섯의 안정적 생산을 위한 최적배지의 개발)

  • Gal, Sang-Wan;Lee, Sang-Won
    • Applied Biological Chemistry
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    • v.45 no.2
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    • pp.71-76
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    • 2002
  • Several antagonistic bacteria, SD-1, 4, 10, 11, 14, 15, and 16, which have strong CMCase and amylase activities, were isolated from the fermented mushroom media. Among them, SD-1, 10, 11, and 15 have strong antibacterial activities against the mushroom pathogenic bacteria Pseudomonas sp., and SD-1, 10, 11, 14, and 16 have strong antifungal activities against the mushroom pathogenic fungi, Trichoderma sp. SD-14, 15, and 16 did not inhibit the growth of mushroom Pleurotus eryngii ASI-2302, and Pleurotus ostreatus ASI-2042 and ASI-2180. When the culture broth mixture of the seven bacterial strains was applied to the mushroom media, the growths of pathogens, Pseudomonas sp. and Trichoderma sp., were inhibited.

Diversity, Saccharification Capacity, and Toxigenicity Analyses of Fungal Isolates in Nuruk (누룩곰팡이 분리균의 다양성 및 당화능 분석과 독소생산능 조사)

  • Kim, Min Sik;Kim, Sinil;Ha, Byeong-Seok;Park, Hye-Young;BaeK, Seong-Yeol;Yeo, Soo-Hwan;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.3
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    • pp.191-200
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    • 2014
  • Nuruk samples collected from various regions in Korea were investigated in terms of fungal contents and diversity. In measurement of colony forming unit (CFU) in Nuruk suspensions on DRBC agar, Nuruk samples MS4, MS8, and MS10 were among the highest fungal density, with $1,278.9{\pm}21.6$ (${\times}10^4$), $1,868.0{\pm}27.7$ (${\times}10^4$), and $775.1{\pm}19.2$ (${\times}10^4$) were among the samples showing the highest fungal density. CFU per 20 mg Nuruk, respectively. The majority of fungal components were yeasts, including Pichia anomala, P. kudriavzevii, Kluyveromyces marxianus, and Saccharomycopsis fibuligera, whereas Aspergillus oryzae and Rhizopus oryzae, the representative Nuruk fungi, were predominant only in the low fungal density Nuruks (MS2, MS5, and MS11). Saccharification capability of the fungal isolates was assessed by measurement of amylase activity in the culture broth. The highest amylase activity was found in A. niger and A. luchuensis, followed by S. fibuligera. A. oryzae and R. oryzae showed fair amylase activity but significantly lower than those of the three fungal species. R. oryzae was suggested to play an additional role in degradation of ${\beta}$-glucan in crop component of Nuruk since R. oryzae was the only fungus that showed ${\beta}$-glucanase activity among the fungal isolates. To confirm the safety of Nuruk, aflatoxigenicity of the isolated Aspergillus was estimated using the DNA markers norB-cypA, aflR, and omtA. All of the isolates turned out to be non-aflatoxigenic as evidenced by the deletion of gene markers, norB-cypA and aflR, and the absence of aflatoxin in the culture supernatants shown by TLC analysis.

Study on Soluble Concentrate Formulation and Quality Control Techniques of a Microbial Insecticide "Bt-Plus" (미생물살충제 "비티플러스" 액상 제형화 및 품질 분석 기술에 관한 연구)

  • Eom, Seonghyeon;Park, Hyeonji;Kim, Kyusoon;Hong, Youkyeong;Park, Jiyeong;Choi, Bongki;Kim, Joonsung;Kim, Kunwoo;Kang, Moonsoo;Yang, Kyunghyung;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.52 no.2
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    • pp.115-123
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    • 2013
  • A microbial insecticide "Bt-Plus" has been developed to enhance an insecticidal efficacy of an entomopathogenic bacterium, Bacillus thuringiensis (Bt). However, its wettable powder formulation is not preferred by farmers and industry producers due to relatively high cost. This study aimed to develop a soluble concentrate formulation of Bt-Plus. To this end, an optimal mixture ratio of two bacterial culture broths was determined to be 5:4 (v/v) of Bt and Xenorhabdus nematophila (Xn) along with 10% ethanol preservative. In addition, Bt broth was concentrated by 10 times to apply the mixture at 1,000 times fold dilution. The resulting liquid formulation was sprayed on cabbage crop field infested by late instar larvae of the diamondback moth, Plutella xylostella. The field assay showed about 77% control efficacy at 7 days after treatment, which was comparable to those of current commercial biopesticides targeting P. xylostella. For storage test in both low and room temperatures, the liquid formation showed a relatively stable control efficacy at least for a month. To develop a quality control technique to exhibit a stable control efficacy of Bt-Plus, Bt spore density ($5{\times}10^{11}$ spores/mL) and eight active component concentrations of Xn bacterial metabolites in the formulation products have been proposed in this study.

Effect of Oxytetracycline Injection an the Body of Paralichthys Olivaceus (Oxytetracycline의 투여방법에 따른 넙치(Paralichthys olivaceus) 체내의 잔류 특성)

  • Ko, Chang-Sik;Kim, Dong-Hwi;Park, So-Hyun;Moon, Kyung-Mi;Heo, Moon-Soo
    • Journal of Life Science
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    • v.26 no.3
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    • pp.309-317
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    • 2016
  • Industrial advancements have resulted in food culture development, followed by increased seafood consumption and large-scale seafood farming, which has been accompanied by an increased prevalence of fish disease. The antibiotic oxytetracycline (OTC) is commonly used to prevent and treat bacterial diseases in fish. However, overuse of OTC had led to negative aspects. In view of this, we conducted a research with regard to aspects of remnants on olive flounder skin, liver, and muscle through dipping treatment and oral feeding of OTC and analyzed the results with bioassay and HPLC quantitative analyses. The dipping treatment was carried out once with 25 g/ton/hr of OTC, and the oral treatment with 62.5 mg/kg body weight/7 days. The results underwent a bioassay analysis. The dipping group reacted only on the skin right after dipping, while the oral feeding group responded on the skin for 77 days after feeding and on the muscle for 14 days. In the dipping group, the HPLC quantitative analysis revealed remnants in the skin on the 37th day and on the 13th day in the liver group. No remnants were found in the muscle, even immediately after dipping. In the oral feeding group, there was a high concentration (1.07 mg/kg) of remnant in the skin, even on the 77th day. 0.56 mg/kg in the liver, even a small amount, and no remnant in the muscle on the 42nd day. To sum up, the results suggest that it will not be harmful to our body to observe the OTC withdrawal period of 40 days with the muscle because OTC will hardly remain on it. When using olive flounder for sashimi, the skin and liver should not be used for broth, as the quantity of OTC residue is several times higher than that found in muscle. As previous studies reported that the concentration of remnants gradually decreased with heating, so it was likely to lessen, depending on the cooking temperature.

Purification and Characterization of $\beta$-Cyclodextrin Glucanotransferase Excreted by Bacillus firmus var. aikalophilus. (호알칼리성 Bacillus firmus가 생산하는 $\beta$-Cyclodextrin Glucanotransferase의 정제 및 효소반응 특성)

  • Shin, Hyun-Dong;Kim, Chan;Lee, Yong-Hyun
    • Microbiology and Biotechnology Letters
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    • v.26 no.4
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    • pp.323-330
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    • 1998
  • Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0~9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

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