• Title/Summary/Keyword: bioassay test

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Ecological Risk Assessment(ERA) of Abandoned Mine Drainage(AMD) in Korea Based on Vibrio fisheri, Selenastrum capricornutum, and Daphnia magna (국내 폐광산 지역의 Vibrio fisheri, Selenastrum capricornutum, 그리고 Daphnia magna를 이용한 생태 위해성 평가)

  • Kim, Ki-Tae;Lee, Byoung-Cheun;Kim, Dong-Wook;Kim, Sang-Don
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.2
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    • pp.163-168
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    • 2007
  • Ecological risk assessment(ERA) to 5 abandoned mine drainage was investigated by using chemical measurement and bioassay experiment. From the results of chemical analysis, the high concentration of heavy metals are detected in most area. The Arsenite were mostly detected in Songcheon, Nakdong, and Dukum abandoned mine area, and various heavy metals were highly dispersed in Nakdong area. The study area have also high biological toxicity, resulted from the bioassay based on WET(Whole Effluent Toxicity) test by using Vibrio fisheri, Selenastrum copricornutum, and Daphnia magna. The maximum toxicity was shown in the point where the mine waters start to flow. The sensitivity of toxicity by S. capricornutum was relatively high considering the values of toxicity in all samples, from 1.3 to 32.0 TU. The different sensitivities of toxicity recommends the use of battery system, resulted from at least two test species for bioassay or ecological risk assessment of mine drainage. Besides, the results showed high hazard quotient(i.e., greater than 1 HQ value indicating potentially significant toxic risks) with regard to abandoned mine drainage area in this study. On the other hand, the biological toxicity results were sharply decreased by attenuation along further distance from discharging of mine waters. Therefore, environmental parameters including the dilution factor, dissolved organic matter, and hardness should be considered when the remediation and ERA of abandoned mine drainage is planned.

Reoxygenation Stimulates EDRE(s) Release from Endothelial Cells of Rabbit Aorta

  • Suh, Suk-Hyo;Han, Jae-Jin;Park, Sung-Jin;Choi, Jai-Young;Sim, Jae-Hoon;Kim, Young-Chul;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.4
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    • pp.393-404
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    • 1999
  • We have reported that hypoxia stimulates EDRF(s) release from endothelial cells and the release may be augmented by previous hypoxia. As a mechanism, it was hypothesized that reoxygenation can stimulate EDRF(s) release from endothelial cells and we tested the hypothesis via bioassay experiment. In the bioassay experiment, rabbit aorta with endothelium was used as EDRF donor vessel and rabbit carotid artery without endothelium as a bioassay test ring. The test ring was contracted by prostaglandin $F_{2a}\;(3{\times}10^{-6}\;M)$ which was added to the solution perfusing through the aorta. Hypoxia was evoked by switching the solution aerated with 95% $O_2/5%\;CO_2$ mixed gas to one aerated with 95% $O_2/5%\;CO_2$ mixed gas. Hypoxia/reoxygenation were interexchanged at intervals of 2 minutes (intermittent hypoxia). In some experiments, endothelial cells were exposed to 10-minute hypoxia (continuous hypoxia) and then exposed to reoxygenation and intermittent hypoxia. In other experiments, the duration of reoxygenation was extended from 2 minutes to 5 minutes. When the donor aorta was exposed to intermittent hypoxia, hypoxia stimulated EDRF(s) release from endothelial cells and the hypoxia-induced EDRF(s) release was augmented by previous hypoxia/reoxygenation. When the donor aorta was exposed to continuous hypoxia, there was no increase of hypoxia-induced EDRF(s) release during hypoxia. But, after the donor aorta was exposed to reoxygenation, hypoxia-induced EDRF(s) release was markedly increased. When the donor aorta was pretreated with nitro-L-arginine $(10^{-5}$ M for 30 minutes), the initial hypoxia-induced EDRF(s) release was almost completely abolished, but the mechanism for EDRF(s) release by the reoxygenation and subsequent hypoxia still remained to be clarified. TEA also blocked incompletely hypoxia-induced and hypoxia/reoxygenation-induced EDRF(s) release. EDRF(s) release by repetitive hypoxia and reoxygenation was completely blocked by the combined treatment with nitro-L-arginine and TEA. Cytochrome P450 blocker, SKF-525A, inhibited the EDRF(s) release reversibly and endothelin antgonists, BQ 123 and BQ 788, had no effect on the release of endothelium-derived vasoactive factors. Superoxide dismutase (SOD) and catalase inhibited the EDRF(s) release from endothelial cells. From these data, it could be concluded that reoxygenation stimulates EDRF(s) release and hypoxia/reoxygenation can release not only NO but also another EDRF from endothelial cells by the production of oxygen free radicals.

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The Study on the Analysis Method of Tetrodotoxin in Puffer Fish (복어 중 테트로도톡신 분석법에 관한 연구)

  • Kang, Young-Woon;Lee, Yoon-Suk;Park, Sung-Kug;Seo, Jung-Heok;Kim, Mee-Hye
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.37-41
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    • 2012
  • The current standard for testing tetrodotoxin (TTX) in foodstuffs is the mouse bioassay (MBA) in Korea as in many other countries. However, this test suffers from potential ethical concerns over the use of live animals. In addition, the mouse bioassay does not test for a specific toxin thus a sample resulting in mouse incapacitation would need further confirmatory testing to determine the exact source toxin (e.g., TTX, STX, brevotoxin, etc.). Furthermore, though the time of death is proportional to toxicity in this assay, the dynamic range for this proportional relationship is small thus many samples must be diluted and new mice be injected to yield a result that falls within the quantitative dynamic range. Therefore, in recent years, there have been many efforts in this field to develop alternative assays. High performance liquid chromatography (HPLC) coupled with mass spectrometry (MS) has been emerged as one of the most promising options. A LC-MS-MS method involves solid-phase extraction (SPE) and followed by analysis using an electrospray in the positive ionization mode and multiple reactions monitoring (MRM). To adopt LC-MS-MS method as alternative standard for testing TTX, we performed a validation study for the quantification of TTX in puffer fish. This LC-MS-MS method showed good sensitivity as limits of detection (LOD) of $0.03{\sim}0.08{\mu}g/g$ and limits of quantification (LOQ) of $0.10{\sim}0.25{\mu}g/g$. The linearity ($r^2$) of tetrodotoxin were 0.9986~0.9997, the recovery were 80.9~103.0% and the relative standard deviations (RSD) were 4.3~13.0%. The correlation coefficient between the mouse bioassay and LC/MS/MS method was higher than 0.95.

Isolation and identification of Actinomycetes for the Control of Agricultual Pests and Fungal Pathogene (농작물 해충 및 진균류 방제를 위한 방선균의 분리 및 동정)

  • 이은정;강경돈
    • Journal of Sericultural and Entomological Science
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    • v.40 no.1
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    • pp.63-69
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    • 1998
  • Twenty seven out of ca. 5,000 actinomycete strains, which were isolated from soil collected throughout the country, showed antibicrobial effects against fungai, Rhizopus stronifer (ATCC 6227a), Rhizoctonia solani (KCCM 11271) and yeast, Candida albicans (ATCC 10231). From these antifungal microorganisms, we further selected seven strains which seemed to produce insecticidal substances with in vivo test, using silkworm, Bombyx mori and beet armyworm, Spodoptera exigua. Morphological and biochemical experiments revealed that three strains out of seven were Streptomyces. Further investigations on the physical and chemical properties of these antifungal and insecticidal substances are now in progress.

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Allelopathy of Tagetes minuta L. Aqueous Extracts on Seed Germination and Root Hair Growth

  • Kil, Ji-Hyun;Shim, Kew-Cheol;Lee, Kyu-Jin
    • The Korean Journal of Ecology
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    • v.25 no.6
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    • pp.395-398
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    • 2002
  • Present paper showed allelopahtic effects of Tagetes minuta aqueous extracts on seed germination and root hair development. Allelopathy of aqueous extracts derived from T. minuta examined using two test plant species (Lotus comiculatus var. japonicus and Lactuca sativa). The seeds of test species were inoculated in petri dishes containing 0, 10,50 and 100% aqueous extracts from T. minuta. At day 5, the relative seed germination ratio to control was evaluated, and the development of seedling root hairs was observed through light microscopy. Seed germination of L. comiculatus var. japonicus was significantly inhibited proportional to the concentrations of aqueous extract, but that of L. sativa wasn't inhibited. The inhibitory allelopathic effect of T. minuta was found in the development and growth of seedling root hairs. It was concluded that the inhibitory allelophatic effects have been to be investigated using various bioassay, for the allelopathy of plant species shows species-specific and organ-specific.

An Assay Method with Lamina Joints of Korean Rice (우리나라 벼의 Lamima Joint를 이용한 생물검정법)

  • Park, Keun-Hyung;Hyun, Kyu-Hawn;Kim, Dong-Youn
    • Applied Biological Chemistry
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    • v.29 no.1
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    • pp.22-28
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    • 1986
  • To establish the bioassay system not only for brassinosteroids and auxins but also for growth retardants with the lamina joints of rice, excellent domestic cultivars were selected and affectable factors, condition of test material, pH, temperature, concentration of test solution, coexisting metallic ions ana combination with growth regulators, on the assay system were discussed.

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The Controlling Activity of Several Fungicides against Rice Bakanae Disease Caused by Fusarium fujikuroi in Five Assay Methods (몇 가지 살균제의 벼 키다리병과 병원균에 대한 효과 검정)

  • Shin, Myeong-Uk;Lee, Su-Min;Lee, Yong-Hwan;Kang, Hyo-Jung;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.12 no.2
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    • pp.168-176
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    • 2008
  • Recently damage of rice bakanae disease disseminated by infected seeds increased in paddy field in Korea. For controlling rice bakanae disease, the efficacy of 17 fungicides was assessed by 5 kinds of bioassay, spore germination test (SGT), mycelial growth test, detection test on Komada's medium (KDT), pouch test (PT) and greenhouse test (GT). Among ergosterol biosynthesis inhibiting fungicides, prochloraz showed a high controlling activity in all the assay systems while the others showed very low activity except for $500\;{\mu}g/ml$ of hexaconazole in GT and $500\;{\mu}g/ml$ of triflumizole in KDT. Although benomyl and the mixture of benomyl and thiram showed a good activity at 100 and $500\;{\mu}g/ml$ in SGT and PT, respectively, in GT they did a middle activity. Trifloxystrobin and kresoxim-methyl included in strobilurins showed a good activity even at $20\;{\mu}g/ml$ in KDT as well as a middle activity in SGT. Also a high activity not only at $10\;{\mu}g/ml$ in SGT but also at $100\;{\mu}g/ml$ KDT was detected in thiram. The activity of fludioxonil was confirmed in SGT, KDT and PT. Based on these results, it is very important to determine a bioassay system, because the fungicidal activity against rice bakanae disease was fluctuated depending on a assay systems as well as the mechanism of fungicide.

Mutagenicity of Diesel Exhaust Emission by Micronucleus Test (디젤엔진 배기물질의 변이원성 연구)

  • 임국환;배은상;김영환;김광종
    • Journal of Environmental Health Sciences
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    • v.17 no.1
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    • pp.13-19
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    • 1991
  • Interest in the studies of diesel exhaust emission has been increasing by the expected increase in the use of diesel powered automobiles out of concideration of fuel economy. It was well known that diesel exhaust emission was mutagenic in the bioassay as Ames test. The authors tried to find out the cytogenetic effect of diesel exhaust emission by the operational condition of engine such as speed and load. For the investigation of those effects, 66 male mice of ICR strain were used. The benzene-ethanol extracts of diesel exhaust emission were injected intra peritoneum 25rng/kg and 50mg/kg respectively. To evaluate the cytogenetic effect, mouse bone marrow micronucleus test was carried out. The frequency of micronucleus was different among the various groups according to the operational conditions of engine. The frequency of micronucleus in idling group was the highest of all the groups the subgroup of 50mg/kg showed the rate of 1.30%, 25rng/kg subgroup 0.55%. And the group of 2000rpm with 50% load showed the lowest rate of micronucleus appearance as 0.20% and 0.15%. In general, the frequency of micronucleus was shown higher in propotion to load and was shown inversely proportional to speed.

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Assays of Residual Antibiotics after Treatment of γ-ray and UV Irradiation (감마선과 UV 조사에 의한 항생제 분해)

  • Shin, Ji-Hye;Nam, Ji-Hyun;Yu, Seungho;Lee, Myunjoo;Lee, Dong-Hun
    • Journal of Radiation Industry
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    • v.4 no.1
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    • pp.39-45
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    • 2010
  • The pollution of antibiotics is a major cause of spreading antibiotics resistant bacteria in the environment. Applications of ozonation, UV, and ${\gamma}-ray$ irradiations have been introduced to remove antibiotics in the effluents from wastewater treatment system. In this study, we compared the chemical (HPLC) and biological (antimicrobial susceptibility test, AMS) assays in measuring of the concentrations of residual antibiotics after ${\gamma}-ray$ and UV irradiation. Most samples were degraded by ${\gamma}-ray$ irradiation (1~2 kGy). However, lincomycin and tetracycline were not degraded by UV irradiation. The concentration of residual antibiotics, that was treated with ${\gamma}-ray$ and UV irradiation, measuring by bioassay was similar to HPLC. The concentrations of ${\gamma}-ray$ irradiated cephradine measured by AMS test were 2 times higher than that of HPLC assay, indicating AMS test is more sensitive than HPLC assay. These results indicate that ${\gamma}-ray$ irradiation technique is more useful than UV irradiation, and biological assay is more useful to detect the antibiotics and toxic intermediates in antibiotics degradation.

Simplified the Screening and In Vitro Appraisal of Antioxidant, Cytotoxic, Thrombolytic, Antimicrobial and Membrane Stabilizing Activities of Lablab Purpures at a Time

  • Rahman, M. Saifur;Uddin, M. Gias;Alam, M. Badrul;Yoo, Jin Cheol
    • Journal of Integrative Natural Science
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    • v.7 no.3
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    • pp.173-182
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    • 2014
  • To simplify the different biological investigation of the methanolic extract and solvent-solvent partitioning of Lablab purpures (L. purpures) bark. In-vitro anti-oxidant study was determined using total DPPH radical scavenging assay. In vitro antimicrobial study was measured by observing zone of inhibition. The cytotoxic activity was studied using brine shrimp lethality bioassay and thrombolytic activity by clot disruption method. The antioxidant potential was evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Folin-Ciocalteau reagents using butylated hydroxytolune (BHT) and ascorbic acid as standards. The Aqueous soluble fraction revealed the highest free radical scavenging activity ($IC_{50}=48.76{\mu}g/mL$). The antimicrobial screening of the bark of L. purpures exhibited mild to moderate activity in test microorganisms. The CSF showed the maximum relative percentage inhibition against Salmonella parathyphi (34.2%) for bacteria and C. albicans (28.8%) for fungi whereas, lowest relative percentage inhibition against Sarcina lutea (22.0%) for bacteria and Aspergillus niger (24.4%) for fungi. In the brine shrimp lethality bioassay, The $LC_{50}$ values of Carbon tetrachloride and N-Hexane soluble fraction were found $92.18{\mu}g/mL$, and $68.95{\mu}g/mL$ respectively while the $LC_{50}$ values of standard Vincristine sulphate was $1.37{\mu}g/mL$. The methanolic extract and its organic soluble fractions of Lablab purpureus at concentration 2.0 mg/mL, significantly protected the lysis of erythrocyte membrane induced by hypotonic solution and heat as compared to the standard, acetyl salicylic acid (0.10 mg/mL). The MSF and AQSF produced 61.48 % and 53.75% inhibition of hemolysis of RBC caused by hypotonic solution respectively, whereas acetyl salicylic acid (0.10 mg/mL) showed 76.42%. Ethanol extract of L. purpures and all of its different partitions exhibited moderate thrombolytic activity of 37.25%-2.40%. Very good preliminary screening and simplified experiments were able to show the different biological activity of methanolic extract and its soluble fractions of L. purpures at a time.