• 제목/요약/키워드: resazurin

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Inhibition of Pathogenic Bacteria and Fungi by Natural Phenoxazinone from Octopus Ommochrome Pigments

  • Lewis-Lujan, Lidianys Maria;Rosas-Burgos, Ema Carina;Ezquerra-Brauer, Josafat Marina;Burboa-Zazueta, Maria Guadalupe;Assanga, Simon Bernard Iloki;del Castillo-Castro, Teresa;Penton, Giselle;Plascencia-Jatomea, Maribel
    • Journal of Microbiology and Biotechnology
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    • 제32권8호
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    • pp.989-1002
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    • 2022
  • Cephalopods, in particular octopus (Octopus vulgaris), have the ability to alter their appearance or body pattern by showing a wide range of camouflage by virtue of their chromatophores, which contain nanostructured granules of ommochrome pigments. Recently, the antioxidant and antimicrobial activities of ommochromes have become of great interest; therefore, in this study, the pH-dependent redox effect of the extraction solvent on the antioxidant potential and the structural characterization of the pigments were evaluated. Cell viability was determined by the microdilution method in broth by turbidity, MTT, resazurin, as well as fluorescence microscopy kit assays. A Live/Dead Double Staining Kit and an ROS Kit were used to elucidate the possible inhibitory mechanisms of ommochromes against bacterial and fungal strains. The results obtained revealed that the redox state alters the color changes of the ommochromes and is dependent on the pH in the extraction solvent. Natural phenoxazinone (ommochromes) is moderately toxic to the pathogens Staphylococcus aureus, Bacillus subtilis, Salmonella Typhimurium and Candida albicans, while the species Pseudomonas aeruginosa and Pseudomonas fluorescens, and the filamentous fungi Aspergillus parasiticus, Alternaria spp. and Fusarium verticillioides, were tolerant to these pigments. UV/visible spectral scanning and Fourier- transform infrared spectroscopy (FTIR) suggest the presence of reduced ommatin in methanol/ HCl extract with high intrinsic fluorescence.

체세포수(Somatic Cell Counts)를 주로한 원유질의 평가 -원유등급제도에 의한 유질향상과 산유량 증가방안- (Analysis if Somatic Cell Counts of Raw Milk in Korea -Recommendation to Payment for Milk on the Basis of Quality-)

  • 손봉환;강구식
    • 한국동물위생학회지
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    • 제14권2호
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    • pp.87-103
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    • 1991
  • The somatic cell counts SCC and bacteria counts were done by D milk plant, P milk plant, S milk plant and Inch'${\v{\times}}n$ Vet. Serv. Lab from 1987 to 1990 with Coulter counter, Fossomatic 90, Bactoscan, Rolling ball viscometer and Resazurin reduction test. The results were summarized as follows 1. In the distribution of SCC of the bulk herd milk, D milk plant from Nov. 1989 to Oct. 1990 remarks 80.2% on the range below 500, 000, 14.5% ranging from 1, 000, 000 to 1, 500, 000, 1.2% ranging from 1, 500, 000 to 2, 000, 000, 0.69% ranging from 2, 000, 000 to 3, 000, 000, 0.71% on the range over 3, 000, 000. P milk plant remarks 237, 000 in the first half year and 251, 000 in the second half year in 1990 year. S milk plant remarks annual average of 335, 000 in 1987, 273, 000 in 1988 and 262, 000 in 1989. The individual record of Inch'${\v{\times}}n$ Vet. Serv Lab. remarks 79.35% and 80.2% below 500, 000 8.30% and 7.40% from 500, 000 to 1, 000, 000, 2.37% and 3.2% from 1, 000, 000 to 1, 500, 000, 2.77% and 2.30% from 1, 500, 000 to 2, 000, 000, 1.67% and 2.00% from 2, 000, 000 to 3, 000, 000, 5.53% and 4.40% over 3, 000, 000 in 1989 and 1990, respectively. The grade distirbution of SCC is as follows: D milk plant shows 1st grade-80.20%, 2nd grade-l6.5% and 3rd grade-3.30%. And P milk plant shows all 1st grade. S milk plant shows 87.30%, 8.6% and 4.1% in 1987 and 91.90%, 6.1% and 2.0% in 1988, and 92.40%, 6.1% and l.5% in 1989 on the 1st, 2nd and 3rd grade respectively. 2. The distribution of bacteria P milk plant reached 15.123 in 1st half year and 21.515 in 2nd half year. Also, S milk plant reached 81.5%, 12.5%, 6.0% in 1987, and 86.20%, 9.70%, 4.1% in 1988, and 86.2%, 10.8%, 3.0% in 1989 respectively for 1st, 2nd and 3rd grade. 3. The regional SCC distribution in D milk plant shows 1, 540, 000 in three regions and 714, 000 in one region. And monthly SCC distribution shows 671, 000 in December and 1, 165, 000 in June. 4. As a result of the individual SCC test, 9 times for 16 cows in “I”farm(1986-1988), and 6 times for 13 cows in“D”farm(1987-1988) No.3, 5, 9, 14 cows in“I”farm showed the high SCC beyond 1, 000, 000 over 4-5times. 5. If the SCC over 300, 000 reach 40%, the national producing quality of milk can be reduced by 87, 600M /I annually and in the sum of money, it should be about 35.5 billion Won. 6. The difference between high group and low group for SCC in D milk plant reached over 1, 000, 000. In case that the difference reaches 1, 000, 000 in the farm bulk milk at a farm breeding 20 cows which produce 20kg milk per day, it was estimate that the annual difference of producing quantity and sum of money respectively should be reached 26, 280kg in milk and 10, 643, 400 Won in income.

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로즈마리 추출물 및 분획물의 항균·항산화 활성 (Antioxidative and Antimicrobial Activities of Methanol Extract from Rosmarinus officinalis L. and Their Fractions)

  • 유미희;채인경;정영태;정연섭;김혁일;이인선
    • 생명과학회지
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    • 제21권3호
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    • pp.375-384
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    • 2011
  • 허브류의 하나인 로즈마리를 추출, 분획하여 각 용매별 항산화와 항균효과를 관찰하였다. 로즈마리의 항산화효과를 측정하기 위해 DPPH radical 소거능과, ABTS radical 소거능 및 FRAP 활성을 측정하였다. DPPH radical 소거능을 측정한 결과 ethyl acetate분획물의 $RC_{50}$값이 $3.22\;{\mu}g/ml$로 강력한 항산화능을 나타냈으며, water 분획물을 제외한 나머지 분획물(methanol: $5.05\;{\mu}g/ml$, hexane: $6.28\;{\mu}g/ml$, chlorofrom: $5.32\;{\mu}g/ml$, buthanol: $5.74\;{\mu}g/ml$)에서도 강력한 항산화능을 보였다. FRAP활성 또한 ethyl acetate 분획물이 $5.9{\pm}0.3\;{\mu}M/{\mu}g$의 높은 $Fe^{2+}$함량을 나타났으며 water 분획물을 제외한 나머지 분획물에서 높은 $Fe^{2+}$함량을 나타내어 DPPH radical 소거능과 유사한 경향을 보였다. ABTS radical 소거능은 ethyl acetate분획물의 $RC_{50}$값이 $5.05\;{\mu}g/ml$, methanol 추출물의 $RC_{50}$값이 $8.72\;{\mu}g/ml$으로 강력한 항산화능을 나타냈으며 그 외 나머지 분획물들은 DPPH radical 소거능보다 상대적으로 낮은 항산화능을 나타냈다. 로즈마리 단일물질로는 rosmarinic acid, carnosic acid가 DPPH radical 소거능과 FRAP활성에 유사한 경향을 보인 반면 ABTS radical 소거능은 rosmarinic acid만이 $43.16\;{\mu}M$에서 $RC_{50}$값을 나타내었다. 로즈마리의 항균 활성은 Paper disc method를 이용한 Inhibition zone과 MIC (최소 저해 농도)를 측정하여 항균력을 알아본 후 hand plate와 resazurin 환원 실험을 통해 항균제와 식품 보존제로서의 이용 가능성을 알아보았다. 항균 활성 검색 결과 methanol 추출물과 chloroform 분획물이 S. aureus에 대해서 2.5 mg/disc의 농도에서 inhibition zone이 각각 15.3 mm, 15.5 mm로 높게 나타났으며 E. coli에 대해서 14.3 mm, 13.7 mm로 나타나 강력한 항균 효과를 나타내었다. MIC는 S. aureaus 균주와 E. coli 균주 모두 methanol 추출물에서 5 mg/ml의 MIC값을 나타내었고 chloroform 분획물은 2 mg/ml에서 MIC값을 나타내었다. 항균활성의 경우 methanol 추출물과 소수성의 hexane, chloroform 분획물에서 높은 활성을 보였다. 위의 결과를 통해 항균제와 식품 보존제 등의 상품화는 경제적이고 그 활성이 우수한 methanol 추출물의 이용이 가장 이상적인 것으로 판단되어 methanol 추출물을 이용하여 항균제와 식품 보존제로의 이용 가능성을 측정한 결과 항균제로는 hand plate를 통해 5 mg/ml에서 식품 보존제로는 resazurin 환원 실험을 통해 1 mg/ml에서 이용이 가능하리라 생각된다.

Screening the extracts of the seeds of Achillea millefolium, Angelica sylvestris and Phleum pratense for antibacterial, antioxidant activities and general toxicity

  • Sarker, Satyajit Dey;Eynon, Elaine;Fok, Katharine;Kumarasamy, Yashodharan;Murphy, Eavan Marie;Nahar, Lutfun;Shaeen, Ehab Mohammed;Shaw, Nichola Mary;Siakalima, Munachonga
    • Advances in Traditional Medicine
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    • 제3권3호
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    • pp.157-162
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    • 2003
  • Various extracts of higher plants have been used in traditional medicine systems for centuries. While tropical and sub-tropical plants have received considerable attention from the researchers for evaluation of their bioactivity, temeperate plants have always been neglected somewhat. Similarly, seeds of the plants have not been considered seriously compared to other plant parts, e.g. leaves, stems, roots, flowers, etc. as a potential source for biologically active compounds. As part of our on-going evaluation of the extracts of the seeds of temperate plants, especially from Scotland, for biological activity, Achillea millefolium, Angelica sylvestris and Phleum pratense have been chosen for the present study. Both A. millefolium and A. sylvestris are well known for their traditional medicinal uses in Europe and also in the orient, but there is no report on any medicinal properties of P. pratense available to date. Extracts of the seeds of these plants have been assessed for their antioxidant and antibacterial potential and also for general toxicity. Both DCM and MeOH extracts of A. millefolium showed the most significant broad spectrum antibacterial activity among the three plants and inhibited the growth of almost all test strains of bacteria. The DCM extracts of all three species were active against methicillin resistant Staphylococcus aureus (MRSA) and Citrobacter freundii $(MIC=6.25{\times}10^{-1}\;mg/mL)$. While the MeOH extracts of A. millefolium and P. pratense were active against C. freundii, that of P. pratense was also active against MRSA. The MeOH extract of A. sylvestris did not show any antibacterial activity against any of the eight bacterial strains at test concentrations. The MeOH extract of P. pratense showed the most prominent antioxidant activity $(IC_{50}=145\;{\mu}g/ml)$ and there was no antioxidant activity observed with the DCM extract of A. millefolium. The DCM extract of P. pratense was the most toxic $(LC_{50}=20\;{\mu}g/ml)$ among the extracts.

여름철 자연증발시스템(NES)의 腞슬러리 증발효율 평가 (Estimation of Evaporation Rate of Swine Slurry Using the Natural Evaporation System(NES) in summer)

  • 김기연;최홍림;김진길
    • Journal of Animal Science and Technology
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    • 제44권4호
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    • pp.459-474
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    • 2002
  • 본 연구는 실험방법이 간단하고 분석비요이 저렴하여, 단시간내에 세균수를 측정할 수 있는 현장적용 가능한 방법을 확립하기 위하여 냉장우육의 총균수를 표준평판배양법과 배양온도를 25$^{\circ}C$, 30$^{\circ}C$, 35$^{\circ}C$ 별로 레자주린 환원색과 환원시간을 측정하여 상관관계를 산출하였다. 연구결과를 보면, 총균수(25$^{\circ}C$/72시간, Y)와 레자주린 환원시간(X)과의 상관계수는 RRT(Resazurin reducion test) 배양온도가 25$^{\circ}C$와 30$^{\circ}C$, 환원색은 청색에서 분홍색으로 변할 때가 각각 -0.95와 -0.94로서 가장 높았다. 상관계수와 환원시간을 감안할 때 RRT 배양온도는 30$^{\circ}C$가 적합하였으며, 이때 회귀방정식은 Y=-0.4386X + 7.7870이었으며, 돼지고기의 총균수가 $10^2$, $10^3$, $10^4$ cfu/$cm^2$일 경우 검사시간은 13.2시간, 10.9시간, 8.6시간이었다. 총균수(30$^{\circ}C$/72시간, Y)와 레자주린 환원시간(X)과의 상관계수는 RRT 배양온도가 30$^{\circ}C$, 환원색은 청색에서 분홍색으로 변할 때가 -0.93으로서 가장 높았다. 이때 회귀방정식은 Y=0-0.417X + 7.5540이었으며, 돼지고기의 총균수가 $10^2$, $10^3$, $10^4$ cfu/$cm^2$일 경우 검사시간은 13.3시간, 10.9시간, 8.5시간이었다. 총균수(35$^{\circ}C$/48시간, Y)와 레자주린 환원시간(X)과의 상관계수는 RRT 배양온도가 30$^{\circ}C$, 환원색은 청색에서 분홍색으로 변할 때가 -0.93으로서 가장 높았다. 이때 회귀방정식은 Y=-0.3514X + 6.7513이었으며, 돼지고기의 총균수가 $10^2$, $10^3$, $10^4$ cfu/$cm^2$일 경우 검사시간은 13.5시간, 10.7시간, 7.8시간이었다.