• Title/Summary/Keyword: environmental DNA

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Genotoxicity Studies on Carrageenan: Short-term In Vitro Assays

  • Chung, Young-Shin;Eum, Ki-Hwan;Choi, Seon-A;Oh, Se-Wook;Park, Sue-Nie;Yum, Young-Na;Kim, Joo-Hwan;Seo, Young-Rok;Lee, Michael
    • Toxicological Research
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    • v.25 no.1
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    • pp.51-58
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    • 2009
  • Carrageenan is a naturally-occurring sulfated polygalactan which has been widely used in the dairy industry and a gelling agent in non-dairy products. In this study, four short-term in vitro genotoxicity assays were investigated to evaluate the potential genotoxic effects of carrageenan. The mutagenicity of carrageenan was evaluated up to a maximum dose of 5 mg/plate in Ames test. There was no increase in the number of revertant colonies compared to its negative control at any dose in all of strains tested. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. Carrageenan was not considered to be clastogenic in this assay at up to the highest feasible concentration which could be evaluated. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that carrageenan has no genotoxicity potential, although there was a marginal increase in micronuclei frequencies and DNA damage in the respective micronucleus and comet assays. Taken together, our results indicate that carrageenan was not genotoxic based on four in vitro genotoxicity results.

In Vitro Genotoxicity Assessment of a Novel Resveratrol Analogue, HS-1793

  • Jeong, Min Ho;Yang, Kwangmo;Lee, Chang Geun;Jeong, Dong Hyeok;Park, You Soo;Choi, Yoo Jin;Kim, Joong Sun;Oh, Su Jung;Jeong, Soo Kyung;Jo, Wol Soon
    • Toxicological Research
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    • v.30 no.3
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    • pp.211-220
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    • 2014
  • Resveratrol has received considerable attention as a polyphenol with various biological effects such as anti-inflammatory, anti-oxidant, anti-mutagenic, anti-carcinogenic, and cardioprotective properties. As part of the overall safety assessment of HS-1793, a novel resveratrol analogue free from the restriction of metabolic instability and the high dose requirement of resveratrol, we assessed genotoxicity in three in vitro assays: a bacterial mutation assay, a comet assay, and a chromosomal aberration assay. In the bacterial reverse mutation assay, HS-1793 did not increase revertant colony numbers in S. typhimurium strains (TA98, TA100, TA1535 and TA1537) or an E. coli strain (WP2 uvrA) regardless of metabolic activation. HS-1793 showed no evidence of genotoxic activity such as DNA damage on L5178Y $Tk^{+/-}$ mouse lymphoma cells with or without the S9 mix in the in vitro comet assay. No statistically significant differences in the incidence of chromosomal aberrations following HS-1793 treatment was observed on Chinese hamster lung cells exposed with or without the S9 mix. These results provide additional evidence that HS-1793 is non-genotoxic at the dose tested in three standard tests and further supports the generally recognized as safe determination of HS-1793 during early drug development.

Genetic Diversity of Taenia asiatica from Thailand and Other Geographical Locations as Revealed by Cytochrome c Oxidase Subunit 1 Sequences

  • Anantaphruti, Malinee Thairungroj;Thaenkham, Urusa;Watthanakulpanich, Dorn;Phuphisut, Orawan;Maipanich, Wanna;Yoonuan, Tippayarat;Nuamtanong, Supaporn;Pubampen, Somjit;Sanguankiat, Surapol
    • Parasites, Hosts and Diseases
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    • v.51 no.1
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    • pp.55-59
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    • 2013
  • Twelve 924 bp cytochrome c oxidase subunit 1 (cox1) mitochondrial DNA sequences from Taenia asiatica isolates from Thailand were aligned and compared with multiple sequence isolates from Thailand and 6 other countries from the GenBank database. The genetic divergence of T. asiatica was also compared with Taenia saginata database sequences from 6 different countries in Asia, including Thailand, and 3 countries from other continents. The results showed that there were minor genetic variations within T. asiatica species, while high intraspecies variation was found in T. saginata. There were only 2 haplotypes and 1 polymorphic site found in T. asiatica, but 8 haplotypes and 9 polymorphic sites in T.saginata. Haplotype diversity was very low, 0.067, in T. asiatica and high, 0.700, in T. saginata. The very low genetic diversity suggested that T. asiatica may be at a risk due to the loss of potential adaptive alleles, resulting in reduced viability and decreased responses to environmental changes, which may endanger the species.

Recent Development of Rapid and Automation Technology for Food Microbiological Examination

  • Hiroshi Kurata
    • Proceedings of the Korean Society of Food Hygiene and Safety Conference
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    • 1996.06a
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    • pp.33-33
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    • 1996
  • Interests in the field of rapid methods and automation in microbiology have been growing steadily on an international scale in recent years. International meetings concerned this problem have been held in elsewhere in the world countries since the past twenty years. But, unfortunately in the field of microbial examination in food hygiene, this problem have not yet been developed so much as in the field of clinical microbiology. Today, I would like to introduce you here present aspects of rapid and automation technologies, those which are manly carrying in milk and meats industries. My illustration will be given recent improved technologies using automatic apparatus and instruments along with process of microbial count procedure. Recent direct microbiological counting system (ChemeScan \ulcorner) as real time ultrasensitive analysis created by Cheminex Ltd., France is now most evolutional instrument to provide direct microbial counts, down to one cell, within 30 minutes. The results from these evaluations how a good correlation between the ChemScan system and the standard plate count method. This system will be successful application for not only in the field of pharmacology but also food microbiology. In addition, current identification of microbes by sophisticated instruments suitable for food microbiology, one of which Biology is manual system (BIOLOG\ulcorner), provides reference-level capability at a modes price. For the manual system, the color reactions in the microplate are read by eye and manually keyed into personal computer. Species identification appears on the computer screen within seconds, along with biotype patterns, a list of closely related species, and other useful statistics. In present this is useful application for microbial ecology and epidemiological survey. RiboPrinter system newly produced by DuPont is now focusing among microbiologists in the world, and is one of the biggest microbial characterization system using a DNA-based approach. The technology analyzer is bacterial culture for its genetic fingerprint or riboprint pattern. Finally Bio-cellTracer system for automatic measurement of fungal growth and Fukitori-Maseter, a Surface Hygiene Monitoring Kit by using swabe procedure in food processing environment are briefly illustrated in this presentation.

Microbial Community Changes in the Soil of Plastic Film House as Affected by Anaerobic Fermentation of Rice Bran or Wheat Bran (쌀겨와 밀기울의 토양 혐기발효 처리가 시설 재배지 토양의 미생물상에 미치는 영향)

  • Kim, Hong-Lim;Weon, Hang-Yeon;Sohn, Bo-Kyun;Choi, Young-Hah;Kwack, Young-Bum
    • Korean Journal of Soil Science and Fertilizer
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    • v.42 no.5
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    • pp.341-347
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    • 2009
  • Soil microbial community has been changed after the treatment of anaerobic fermentation using wheat bran or rice bran was applied to the soil. In the dilution plate technique, the number of anaerobic bacteria and fungi was higher in rice bran-treated soil than in non and wheat bran-treated soil, but of yeast was higher in wheat bran-treated soil than in non and rice bran-treated soil. Specially, the fungi were not detected in the wheat bran-treated soil. Identified by 16S rDNA sequencing, the number of aerobic bacteria was similar in all treatments, the dominant bacteria was the genus Bacillus. In the phospholipid fatty acid (PLFA) technique, both Gram-positive and Gram-negative bacteria change slightly in all treatments for 20 days of fermentation process but, after 20day, increased rapidly in wheat or rice bran-treated soil. In conclusion, the microbial communities structure was dramatically changed after the treatment of wheat or rice bran to soil.

Development of SSR Markers and Their Use in Studying Genetic Diversity and Population of Finger Millet (Eleusine coracana L. Gaertn.)

  • Lee, Kyung Jun;Yoon, Mun-Sup;Shin, Myoung-Jae;Lee, Jung-Ro;Cho, Yang-Hee;Lee, Ho-Sun;Ma, Kyung-Ho;Lee, Gi-An
    • Plant Breeding and Biotechnology
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    • v.5 no.3
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    • pp.183-191
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    • 2017
  • Finger millet (Eleusine coracana L. Gaertn.) is an important cereal crop in eastern Africa and southern India with excellent grain storage capacity and the unique ability to thrive in extreme environmental conditions. In this study, we analyzed the genetic diversity and population structure of finger millet using 12 developed microsatellites. By sequencing 815 clones from an SSR-enriched genomic DNA library, we obtained 12 polymorphic SSR markers, which also revealed successful amplicons in finger millet accessions. Using the developed SSR markers, we estimated genetic diversity and population structure among 76 finger millet accessions in Asia, Africa, and unknown origins. The number of alleles ranged from 2 to 9, with an average of 3.3 alleles. The mean values of observed heterozygosity and expected heterozygosity were 0.27 and 0.35, respectively. The average polymorphism information content was 0.301 in all 76 finger millet accessions. AMOVA analysis showed that the percentage of molecular variance among the populations was 1%, that among individuals was 5%, and that within individuals was 94%. In STRUCTURE analysis, the 76 finger millet accessions were divided into two subpopulations which had an admixture of alleles. There was a correspondence among PCoA, AMOVA, and population structure. This study may form the basis for a finger millet breeding and improvement program.

Molecular biological analysis of Bt-transgenic (Bt-9) rice and its effect on Daphnia magna feeding

  • Oh, Sung-Dug;Yun, Doh-Won;Chang, Ancheol;Lee, Yu-jin;Lim, Myung-Ho
    • Korean Journal of Agricultural Science
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    • v.46 no.1
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    • pp.113-124
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    • 2019
  • Insect-resistant transgenic (Bt-9) rice was generated by inserting mCry1Ac1, a modified gene from the soil bacterium Bacillus thuringiensis, into the genome of a conventional variety of rice (Ilmi). With regard to potential problems such as safety, an evaluation of non-target organisms is necessary as an essential element of an environmental risk assessment of genetically modified (GM) crops. We studied the effects of the Bt-9 rice on the survival of cantor Daphnia magna, a commonly used model organism in ecotoxicological studies. D. magna fed on the Bt-transgenic rice (Bt-9) and its near non-GM counterparts (Ilmi) grown in the same environment (a 100% ground rice suspension). The Bt-9 rice was confirmed to have the inserted T-DNA and protein expression evident by the PCR and ELISA analyses. The feeding study showed a similar cumulative immobility and abnormal response of the Daphnia magna between the Bt-9 rice and Ilmi. Additionally, the 48 h-EC50 values of the Bt-9 and Ilmi rice were 4,400 mg/L (95% confidence limits: 3861.01 - 5015.01 mg/L) and 5,564 mg/L (95% confidence limits: 4780.03 - 6476.93 mg/L), respectively. The rice NOEC (No observed effect concentration) value for D. magna was suggested to be 1,620 mg/L. We conclude that the tested Bt-9 and Ilmi have a similar cumulative immobility for D. magna, a widely used model organism, and the growth of Bt-9 did not affect non-target insects.

Effects of short-term fasting on in vivo rumen microbiota and in vitro rumen fermentation characteristics

  • Kim, Jong Nam;Song, Jaeyong;Kim, Eun Joong;Chang, Jongsoo;Kim, Chang-Hyun;Seo, Seongwon;Chang, Moon Baek;Bae, Gui-Seck
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.6
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    • pp.776-782
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    • 2019
  • Objective: Fasting may lead to changes in the microbiota and activity in the rumen. In the present study, the effects of fasting on rumen microbiota and the impact of fasting on in vitro rumen fermentation were evaluated using molecular culture-independent methods. Methods: Three ruminally cannulated Holstein steers were fed rice straw and concentrates. The ruminal fluids were obtained from the same steers 2 h after the morning feeding (control) and 24 h after fasting (fasting). The ruminal fluid was filtrated through four layers of muslin, collected for a culture-independent microbial analysis, and used to determine the in vitro rumen fermentation characteristics. Total DNA was extracted from both control and fasting ruminal fluids. The rumen microbiota was assessed using denaturing gradient gel electrophoresis (DGGE) and quantitative polymerase chain reaction. Microbial activity was evaluated in control and fasting steers at various intervals using in vitro batch culture with rice straw and concentrate at a ratio of 60:40. Results: Fasting for 24 h slightly affected the microbiota structure in the rumen as determined by DGGE. Additionally, several microorganisms, including Anaerovibrio lipolytica, Eubacterium ruminantium, Prevotella albensis, Prevotella ruminicola, and Ruminobacter amylophilus, decreased in number after fasting. In addition, using the ruminal fluid as the inoculum after 24 h of fasting, the fermentation characteristics differed from those obtained using non-fasted ruminal fluid. Compared with the control, the fasting showed higher total gas production, ammonia, and microbial protein production (p<0.05). No significant differences, however, was observed in pH and dry matter digestibility. Conclusion: When in vitro techniques are used to evaluate feed, the use of the ruminal fluid from fasted animals should be used with caution.

Description of Diplotriaena manipoli (Nematoda: Diplotriaenoidea) Detected in the Body Cavity of Garrulus glandarius brandtii from Republic of Korea

  • Hong, Eui-Ju;Ryu, Si-Yun;Chae, Joon-Seok;Kim, Hyeon-Cheol;Park, Jinho;Cho, Jeong-Gon;Choi, Kyoung-Seong;Yu, Do-Hyeon;Park, Bae-Keun
    • Journal of Veterinary Clinics
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    • v.36 no.3
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    • pp.133-138
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    • 2019
  • The present study was performed to identify the nematodes recovered from the Eurasian jay, Garrulus glandarius brandtii, from Daejeon Metropolitan City, the Republic of Korea. Total five nematode worms were detected in the body cavities of two out of the twenty birds necropsied, and they were identified using morphological features, light and scanning electron microscope (SEM), and molecular (18S rRNA analysis) methods. The nematodes were all female Diplotriaena manipoli and had numerous eggs at different developmental stages in the uterus. The nematodes were long and slender measuring about 123-145 mm. The eight submedian cephalic papillae were arranged into four large, outer papillae and four small, inner-circle papillae. A pair of amphidal pores were located at the lateral portion of the mouth. The manubrium apex of trident was rounded and three branches of trident were bluntly rounded at the posterior ends. Using 18S rRNA partial sequence arrangements, DNA analysis of nematode worms was also carried out, and they were identified to be close to the Serratospiculum tendo based on a phylogenic tree analysis. To our knowledge, this is the first report on the molecular characterization and SEM study of D. manipoli.

Risk Assessment of Ethylhexyl Dimethyl PABA in Cosmetics

  • Sung, Chi Rim;Kim, Kyu-Bong;Lee, Joo Young;Lee, Byung-Mu;Kwack, Seung Jun
    • Toxicological Research
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    • v.35 no.2
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    • pp.131-136
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    • 2019
  • Ethylhexyl dimethyl para-aminobenzoic acid (PABA) is an oily yellow liquid derivative of water-soluble PABA commonly used in sunscreen. Ethylhexyl dimethyl PABA is widely used as an ingredient in many cosmetics at an average concentration of 1.25% (0.5-2.0%) in Korea. Previous studies, including those involving animals, have demonstrated that ethylhexyl dimethyl PABA is toxic to the following four organs: testis, epididymis, spleen, and liver. In addition, experiments using human keratinocytes found that ethylhexyl dimethyl PABA inhibits cell growth and DNA synthesis at low concentrations, and halted the cell cycle of MM96L cells (human melanoma cell line) at the G1 phase. Despite limited clinical data in humans, many studies have confirmed increased mutagenicity of ethylhexyl dimethyl PABA following exposure to sunlight, which suggests that this molecule is likely to contribute to onset of sun-induced cancer despite protecting the skin through absorption of UVB. For risk assessment, the no observed adverse effect level (NOAEL) chosen was 100 mg/kg bw/day in a 4 weeks oral toxicity study. Systemic exposure dosage (SED) was 0.588 mg/kg bw/day for maximum use of ethylhexyl dimethyl PABA in cosmetics. Based on the risk assessment and exposure scenarios conducted in this study, the margin of safety (MOS) was calculated to be 180.18 for a sunscreen containing 8% ethylhexyl dimethyl PABA, which is the maximum level allowed by the relevant domestic authorities.