• Title/Summary/Keyword: enrichment culture

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Development of a Rapid Enrichment Broth for Vibrio parahaemolyticus Using a Predictive Model of Microbial Growth with Response Surface Analysis (미생물 생장 예측모델과 반응표면분석법을 이용한 Vibrio parahaemolyticus의 신속 증균배지 개발)

  • Yeon-Hee Seo;So-Young Lee;Unji Kim;Se-Wook Oh
    • Journal of Food Hygiene and Safety
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    • v.38 no.6
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    • pp.449-456
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    • 2023
  • In this study, we developed Rapid Enrichment Broth for Vibrio parahaemolyticus (REB-V), a broth capable enriching V. parahaemolyticus from 100 CFU/mL to 106 CFU/mL within 6 hours, which greatly facilitates the rapid detection of V. parahaemolyticus. Using a modified Gompertz model and response surface methodology, we optimized supplement sources to rapidly enrich V. parahaemolyticus. The addition of 0.003 g/10 mL of D-(+)-mannose, 0.002 g/10 mL of L-valine, and 0.002 g/10 mL of magnesium sulfate to 2% (w/v) NaCl BPW was the most effective combination of V. parahaemolyticus enrichment. Optimal V. parahaemolyticus culture conditions using REB-V were at pH 7.84 and 37℃. To confirm REB-V culture efficiency compared to 2% (w/v) NaCl BPW, we assessed the amount of enrichment achieved in 7 hours in each medium and extracted DNA samples from each culture every hour. Real-time PCR was performed using the extracted DNA to verify the applicability of this REB-V culture method to molecular diagnosis. V. parahaemolyticus was enriched to 5.452±0.151 Log CFU/mL in 2% (w/v) NaCl BPW in 7 hours, while in REB-V, it reached 7.831±0.323 Log CFU/mL. This confirmed that REB-V enriched V. parahaemolyticus to more than 106 CFU/mL within 6 hours. The enrichment rate of REB-V was faster than that of 2% (w/v) NaCl BPW, and the amount of enrichment within the same time was greater than that of 2% (w/v) NaCl BPW, indicating that REB-V exhibits excellent enrichment efficiency.

Evaluation of Conventional Culture Methods and Validation of Immunoassays for Rapid Detection of Listeria monocytogenes in Dairy and Processed Foods (유제품 및 가공식품에서 Listeria monocytogenes 검출을 위한 배지법과 신속 검사키트의 유효성 검증)

  • Han, So-Ri;Hyeon, Ji-Yeon;Kim, Hee-Yun;Park, Jong-Seok;Heo, Seok;Shin, Ho-Chul;Seo, Kun-Ho
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.616-622
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    • 2008
  • Listeria monocytogenes is a foodborne pathogen inducing listeriosis in human. We compared two different culture methods for detection of L. monocytogenes and validated two commercial kits, $VIDAS^{(R)}$ and $REVEAL^{(R)}$ for Listeria. L. monocytogenes was inoculated into various food samples to generate partial positive samples. The inoculated samples were enriched in half-Fraser broth for 48 hr at $30^{\circ}C$. The enriched samples were streaked onto Oxford agar at 24 and 48 hr postincubation followed by biochemical confirmation and concurrently analyzed by using the two commercial kits for comparison. When the enrichment period was extended from 24 to 48 hr, the numbers of positive samples were dramatically increased from 6 to 52 out of 80 samples tested using the culture method. With the commercial kits, the numbers of positive samples were also significantly increased from 10 to 18 and 1 to 18, respectively, when the enrichment period was extended from 48 to 72 hr. There was no statistical difference between the 24 hr culture method and $VIDAS^{(R)}$ or $Reveal^{(R)}$ with 48 hr enrichment. In conclusion, the 24 hr for the culture method was insufficient to detect L. monocytogenes in various foods. The commercial kits could be adequate means for presumptive screening of L. monocytogenes in food.

Comparison of Growth Rates of Listeria Interspecies in Different Enrichment Broth (증균배지에서의 Listeria Interspecies의 경쟁생육 비교)

  • Lee, Da Yeon;Cho, Yong Sun
    • Journal of Food Hygiene and Safety
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    • v.33 no.1
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    • pp.65-70
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    • 2018
  • Monitoring of Listeria monocytogenes, the causative agent of listeriosis, in food is inportant for public health. The Korean Food Standards Codex has adopted a 'zero-tolerance' policy for L. monocytogenes. The standard detection method of L. monocytogenes is based on enrichment. Thus, proper enrichment methods need to be instituted to ensure quality control of the detection procedures. In this study, the growth of L. monocytogenes and Listeria innocua as a mixed culture in Listeria enrichment broth (LEB) was monitored during artificial contamination of enrichment culture. We confirmed competitive growth or interspecies inhibitory activity of L. monocytogenes and L. innocua. Interspecies growth differences and the inhibitory activity of different inoculation and mixtures L. innocua against L. monocytogenes were examined. The concentration of L. monocytogenes must be 2.0 log CFU/mL or more than L. innocua to grow better than L. innocua. It is known that Listeria spp. and L. monocytogenes show growth difference during LEB, resulting in the risk of false-negative results. The inhibition of L. monocytogenes by L. innocua was always observed when present at lower concentrations. However, it was confirmed that L. innocua suppressed when L. monocytogenes was present at a higher concentration. Therefore if a mixture of Listeria spp. is present, detecting L. monocytogenes is difficult. Thus, a new enrichment broth to improve the detection rate of L. monocytogenes is needed.

Isolation of an Indigenous Imidacloprid-Degrading Bacterium and Imidacloprid Bioremediation Under Simulated In Situ and Ex Situ Conditions

  • Hu, Guiping;Zhao, Yan;Liu, Bo;Song, Fengqing;You, Minsheng
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1617-1626
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    • 2013
  • The Bacterial community structure and its complexity of the enrichment culture during the isolation and screening of imidacloprid-degrading strain were studied using denaturating gradient gel electrophoresis analysis. The dominant bacteria in the original tea rhizosphere soil were uncultured bacteria, Rhizobium sp., Sinorhizobium, Ochrobactrum sp., Alcaligenes, Bacillus sp., Bacterium, Klebsiella sp., and Ensifer adhaerens. The bacterial community structure was altered extensively and its complexity reduced during the enrichment process, and four culturable bacteria, Ochrobactrum sp., Rhizobium sp., Geobacillus stearothermophilus, and Alcaligenes faecalis, remained in the final enrichment. Only one indigenous strain, BCL-1, with imidacloprid-degrading potential, was isolated from the sixth enrichment culture. This isolate was a gram-negative rod-shaped bacterium and identified as the genus Ochrobactrum based on its morphological, physiological, and biochemical properties and its 16S rRNA gene sequence. The degradation test showed that approximately 67.67% of the imidacloprid (50 mg/l) was degraded within 48 h by strain BCL-1. The optimum conditions for degradation were a pH of 8 and $30^{\circ}C$. The simulation of imidacloprid bioremediation by strain BCL-1 in soil demonstrated that the best performance in situ (tea soil) resulted in the degradation of 92.44% of the imidacloprid (100 mg/g) within 20 days, which was better than those observed in the ex situ simulations that were 64.66% (cabbage soil), 41.15% (potato soil), and 54.15% (tomato soil).

Effects of $\textrm{CO}_2$ Enrichment During Seedling Stage on the Effectiveness of $\textrm{CO}_2$ Enrichment after Transplanting in Leafy Vegetables (엽채류 육묘시 $\textrm{CO}_2$ 시용이 정식 후 $\textrm{CO}_2$ 시용 효과에 미치는 영향)

  • 김일섭;신석범;전익조
    • Journal of Bio-Environment Control
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    • v.11 no.1
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    • pp.35-39
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    • 2002
  • This study was conducted to investigate the effect of early $CO_2$ enrichment during seedling stage on long-term $CO_2$ enrichment after transplanting in the culture of pat-choi (Brassica campesris L), spinach (Spinacia oeracea L.), and leaf lettuce (Lactuca saliva L). During seedling stage, $CO_2$enrichment had significantly higher fresh and dry weight and leaf area of the top parts (above ground) of all three plant species than the control (no $CO_2$ enrichment). About 53%, 70% , and 40% increase in fresh weight of the top parts of pak-choi, spinach, and leaf lettuce were observed, respectively. Also, in seedling stage, dry weights of roots of spinach and leaf lettuce were significantly increased by early $CO_2$ enrichment. Relative fresh weight increment, compared with fresh weight of the control, in the top parts of roll three plants showed the highest values in 10 days after $CO_2$ enrichment treatment. In the long-term $CO_2$ enrichment experiment, early $CO_2$ enrichmented plants had 20% greater leaf area than the control in all three leafy vegetables. Fresh and dry weights of top parts of early $CO_2$-treated plants were also increased from 10 to 20%, as compared with the control plants. However, these increasement rates in the long-term $CO_2$ enrichment were lower than those seedling stage, which had 30-60% increment-rates. After transplanting, photosynthetic rate of each leafy vegetable in the control treatment slowly decreased, but those rates of early $CO_2$ enriched plants rapidly decreased.

Change of fatty acid compositions of rotifer according to enrichment diets and methods in the high density culture (고밀도 배양에 있어서 영양강화 방법 및 종류에 따른 rotifer의 지방산 조성의 변화)

  • PARK Heum Gi;LEE Kyun Woo;LEE Sang-Min;KIM Sung Koo;KIM Hyung Sun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.6
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    • pp.748-752
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    • 1999
  • This study was carried out to compare the growth and fatty acids composition of the rotifer (Brachionus rotundiformis) cultured in high density by the various enrichments and culture methods. The rotifer fed on condensed freshwater Chlorella was enriched with $\omega$-yeast, Algamac, Super Selco and marine Chlorella. In another culture method, the rotifer was cultured with enrichment supplements for 6 hours after feeding with condensed freshwater Chlorella supplement for 18 hour. The rotifer fed with condensed marine Chlorella for 24 hours without fieshwater Chlorella was used as a control group. Culture tanks (5 $\ell$ working volume) was immersed in a water bath ($28^{\circ}C$). The density of rotifer and dissolved oxygen level in water was stable in control group of rotifer cultured with condensed marine Chlorella for 24 hours and the n-3 HUFA content of rotifer was the highest among the rotifer culture methods. However, the density of rotifer and dissolved oxygen level in the groups of rotifers enriched with $\omega$-yeast, Algamac and Super Selco by methods were drastically decreased, The n-3 HUFA contents of rotifers enriched by Super Selco were higher than those of rotifer enriched by either $\omega$-yeast or Algamac in both methods. The results from this experiment indicated that supplementation of condensed marine Chlorella for 24 hour by the semi-continuous culture was effective for the improvement of the nutritional value of rotifer and it could provide the stable growth condition for rotifer culture in high density.

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Micropropagation of Sweetpotato (Ipomoea batatas) in a novel $CO_2$-Enriched Vessel

  • Silva Jaime A. Teixeira da;Giang Dam Thi Thanh;Tanaka Michio
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.67-74
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    • 2005
  • To overcome various disadvantages of conventional cul-ture vessels for micropropagation, a novel disposable vessel, the 'Vitron', made of a multi-layered $OTP^{(R)}$ film and supported by a polypropylene frame, was developed. The film possesses superior properties such as: high light transmittance, low water vapor transmittance and thermal stability and in particular, high gas-permeability. Single nodal explants, which were excised from the multiple shoots derived from shoot-tip culture, were cultured in Vitron and polycarbonate vessels on $3\%$ sugar-containing agar on MS medium and placed at 3000 ppm $CO_2$-enrichment at a low photosynthetic photon flux density (PPFD) ($45{\mu}mol\;m^{-2}\;s^{-1}$). The in vitro and ex vitro growth, and the net photosynthetic rate of in vitro and ex vitro plantlets were significantly enhanced in the Vitron compared to those cultured in a polycarbonate vessel. Explants that were cultured on the same MS medium under low PPFD at various $CO_2$ concentrations were also cultured at 3000 ppm $CO_2$- enrichment at various PPFD: 30, 45, 60, 75 and $90{\mu}mol\;m^{-2}\;s^{-1}$. The best in vitro and ex vitro growth obtained for 3000 ppm $CO_2$-enrichment at $75{\mu}mol\;m^{-2}\;s^{-1}$ PPFD. The novel Vitron vessel, when placed under the two conditions, may replace conventional culture vessels for the successful micropropagation of sweetpotato.

A Functional Matrix Approach to Pedagogical Enrichment of the Dispositional Core of Future Specialists' Experience of Social Interaction

  • Kovalenko, E.V.;Gubarenko, I.V.;Kovalenko, V.I.
    • International Journal of Computer Science & Network Security
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    • v.22 no.11
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    • pp.255-259
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    • 2022
  • The new social reality emerging amid the global rise of communication links and integration processes acutely emphasizes the problems of communication in large and small social systems. The method of their communication becomes one of the keys to ensuring global security. It has become the mission of humanitarian education to prepare the younger generations for life in a changing world with no image of the future and increasing uncertainty. In psychological and pedagogical research, there is a growing scientific interest in the problems of interaction of the individual with the social environment. The mental trace of a person's practice in society shapes the experience of social interaction, which constitutes simultaneously the source, tool, and condition for the emergence and development of personality. The study outlines the methodological foundations for the study of individual experiences of social interaction. A hypothesis about the productivity of the functional matrix method is tested. Materials for the training of specialists in the humanities include interdisciplinary approaches to the study and transformation of the experience of social interaction and systematic methodology for the study of complex objects. Fundamental to the study is the systematic-dialectical method, and the matrix method is employed as the instrumental-technological method. The paper presents the results of a multidisciplinary overview of scientific literature concerning the essential characteristics and functions of social interaction and the respective experience. The overview points to the fragmented nature of scientific understanding of the elements of experience outside its integrity and systemic properties. Based on the formula "personality interacts with the social environment", the study presents an algorithm for the application of a systematic methodology for the study of complex objects, which made it possible to identify the system parameters of experience at three levels of cognition and develop the reference structural and functional matrices for the didactic system of its pedagogical enrichment.

Rapid Detection of Escherichia coli in Fresh Foods Using a Combination of Enrichment and PCR Analysis

  • Choi, Yukyung;Lee, Sujung;Lee, Heeyoung;Lee, Soomin;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Oh, Hyemin;Lee, Yewon;Kim, Yujin;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.38 no.4
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    • pp.829-834
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    • 2018
  • The objective of this study was to determine the minimum enrichment time for different types of food matrix (pork, beef, and fresh-cut lettuce) in an effort to improve Escherichia coli detection efficiency. Fresh pork (20 g), beef (20 g), and fresh-cut lettuce (20 g) were inoculated at 1, 2, and 3 Log CFU/g of Escherichia coli. Samples were enriched in filter bags for 3 or 5 h at $44.5^{\circ}C$, depending on sample type. E. coli cell counts in the samples were enriched in E. coli (EC) broth at 3 or 5 h. One milliliter of the enriched culture medium was used for DNA extraction, and PCR assays were performed using primers specific for uidA gene. To detect E. coli (uidA) in the samples, a 3-4 Log CFU/mL cell concentration was required. However, E. coli was detected at 1 Log CFU/g in fresh pork, beef, and fresh-cut lettuce after 5, 5, and 3-h enrichment, respectively. In conclusion, 5-h enrichment for fresh meats and 3-h enrichment for fresh-cut lettuce in EC broth at $44.5^{\circ}C$, and PCR analysis using uidA gene-specific primers were appropriate to detect E. coli rapidly in food samples.

Reductive Dechlorination of Polychlorinated Biphenyls as Affected by Natural Halogenated Aromatic Compounds

  • Kim Jongseol;Lee Ahmi;Moon Yong-Suk;So Jae-Seong;Koh Sung-Cheol
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.23-28
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    • 2006
  • We investigated the effects of halogenated aromatic compounds (HACs) including naturally occurring ones (L-thyroxine, 3-chloro-L-tyrosine, 5-chloroindole, 2-chlorophenol, 4-chlorophenol and chlorobenzene) on polychlorinated biphenyl (PCB) dechlorination in sediment cultures. A PCB-dechlorinating enrichment culture of sediment microorganisms from the St. Lawrence River was used as an initial inoculum. When the culture was inoculated into Aroclor 1248 sediments amended with each of the six HACs, the extent of dechlorination was not enhanced by amendment with HACs. The dechlorination patterns in the HAC-amended sediments were nearly identical to that of the HAC-free sediments except the 3-chloro-L-tyrosine-amended ones where no dechlorination activity was observed. When these sediment cultures were transferred into fresh sediments with the same HACs, the dechlorination specificities remained the same as those of the initial inoculations. Thus, in the present study, the substrate range of the highly selected enrichment culture could not be broadened by the HACs. It appears that HACs affect PCB dechlorination mainly through population selection rather than enzyme induction of single population.