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The pathogenicity of Streptococcus parauberis isolated from cultured olive flounder Paralichthys olivaceus (양식 넙치, Paralichthys olivaceus에서 분리한 연쇄구균증 원인균, Streptococcus parauberis의 병원성)

  • Choi, Hee-Jung;Cho, Mi-Young;Lee, Jae-Il;Kwon, Mun-Gyeong;Choi, Dong-Lim;Kim, Jin-Woo;Han, Myoung-Chul;Lee, Deok-Chan
    • Journal of fish pathology
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    • v.22 no.3
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    • pp.263-273
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    • 2009
  • There is an increasing number of reports describing Streptococcus parauberis as an important pathogen of cultured olive flounder Paralichthys olivaceus and starry flounder Platichthys stellatus in Korea. We tried to determine the effects of water temperature (14${^{\circ}C}$ and 21${^{\circ}C}$) on the pathogenicity in Streptococcal disease caused by S. parauberis. We have challenged 180 olive flounder by i.p injection to $2.0{\times}10^{7}$ live cells/fish. Mortality was monitored for 21 days post challenge. And histopathological characterizations as infection degree, tissue degeneration and/or bacterial distribution were investigated with H&E stain and in situ hybridization technique. Fifty percent and 16.7% of mortality occurred within 21 days at 21${^{\circ}C}$ and 14${^{\circ}C}$ water temperature, respectively. In most cases, the typical symptoms of olive flounder infected with S. parauberis were darkness of the skin, lethargy, mild abdominal distension cause by ascites, splenomegaly, congested liver and internal organs paleness. The pericardial sac contained large amounts of cloudy fluid. Numerous whitish nodules, which were variable in size and often confluent, were randomly scattered throughout the myocardium. Especially, pericarditis and/or myocarditis was observed in all tested fishes after death. Positive in reaction with S. parauberis were found in all tissues in situ hybridization analysis. The relative numbers of S. parauberis in heart were much more than in liver, spleen, kidney and stomach. We evaluated that S. parauberis strain causes serious damage in the pericardium, shortness of breath and the blood disorder. Therefore, pericarditis and myocarditis caused by S. parauberis were closely related to mortality of olive flounder.

Development of Prevotella nigrescens ATCC $33563^T$-Specific PCR Primers (Prevotella nigrescens ATCC $33563^T$ 균주-특이 중합효소연쇄반응 프라이머 개발)

  • Song, Soo-Keun;Yoo, So-Young;Kim, Mi-Kwang;Kim, Hwa-Sook;Lim, Sun-A;Kim, Do-Kyung;Park, Jae-Yoon;Kook, Joong-Ki
    • Korean Journal of Microbiology
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    • v.44 no.3
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    • pp.212-220
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    • 2008
  • A Pn10 DNA probe was introduced as a Prevotella nigrescens ATCC $33563^T$-specific DNA probe. In that study, the specificity of the Pn10 was tested with only type or reference strains of 5 oral bacterial species. The purpose of this study is to evaluate the specificity of the Pn10 using the wild type strains of P. nigrescens and is to develop the P. nigrescens ATCC $33563^T$-specific PCR primers based on the nucleotide sequence of the Pn10. The specificity of the Pn10 DNA probe was determined by Southern blot analysis. The nucleotide sequence of Pn10 DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pn10 DNA probe were hybridized with the genomic DNAs from P. nigrescens ATCC $33563^T$ and KB6. The Pn10 homologous region, KB6-Pn10, of P. nigrescens KB6 was cloned by PCR and sequenced. The Pn10 and KB6-Pn10 DNA fragments were consisted of 1,875 bp and 1,873 bp, respectively. The percent identity of the two was 98.8% and the divergence of them was 0.6%. The two primer sets (Pn10-F-AC/ Pn10-R-AC and Pn10-F-A/ Pn10-R-A), designed base on the nucleotide sequences of Pn10 DNA probe, were specific to the P. nigrescens ATCC $33563^T$. The two PCR primer sets could detect as little as 4 pg of genomic DNA of P. nigrescens ATCC $33563^T$. These results indicate that the two PCR primer sets have proven useful for the identification of P. nigrescens ATCC $33563^T$, especially with regard to the maintenance of the strain.

Effects of ionizing and ultraviolet radiation on microbial mutation and DNA damage (전리방사선 및 자외선의 미생물 돌연변이와 DNA 손상에 대한 영향)

  • Nam, Ji-Hyun;Shin, Ji-Hye;Lee, Jung-Yun;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.53 no.1
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    • pp.20-28
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    • 2017
  • Physical sterilization methods using ultraviolet radiation and ionizing radiation such as gamma ray and electron beam are applied in various industry fields due to disinfection effects and economic efficiency but may also cause microbial mutation. In this research, Salmonella enterica and Escherichia coli strains were treated with ionizing and ultraviolet radiation and their survival rate, mutation rate, and DNA damage were studied to evaluate the genetic safety. The survival rate of the strains decreased drastically as the irradiation dose of ultraviolet ray, gamma ray, and electron beam increased, and over 90% of the strain was exterminated at a dosage of $0.40{\sim}25.06mJ/cm^3$, 0.11~0.22 kGy, 0.14~0.53 kGy respectively. In SOS / umu-test, genotoxicity causing DNA damage was identified in all samples. In Ames test, back-mutation rate increased to $3.82{\times}10^{-4}$ and $9.84{\times}10^{-6}$ respectively when exposed to ultraviolet ray and gamma ray. At exposure to ultraviolet ray, gamma ray, and electron beam with dosage of over 99.99% extinction rate of S. enterica TA100, back-mutation rate increased 347 times, 220 times, 0.6 times respectively to the spontaneous back-mutation rate. Rifampicin resistance mutation rate of E. coli CSH100 exposed to ultraviolet ray, gamma ray, and electron beam was $2.46{\times}10^{-6}$, $1.66{\times}10^{-6}$, $4.12{\times}10^{-7}$ respectively. Therefore, gamma radiation is effective in microorganism control from the perspective of disinfection and electron beam has the advantage of sterilizing with little DNA damage and bacterial mutation.

Serotype Distribution of Invasive Group B Streptococcal Diseases in Infants at Two University Hospitals in Korea (영아에서의 침습 B군 사슬알균 감염증 및 혈청형 분석: 2개 대학병원 연구)

  • Cho, Hye-Kyung;Nam, Hye Na;Cho, Hye Jung;Son, Dong Woo;Cho, Yong Kyun;Seo, Yiel-Hea;Kim, Yae-Jean;Eun, Byung Wook
    • Pediatric Infection and Vaccine
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    • v.24 no.2
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    • pp.79-86
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    • 2017
  • Purpose: This study was aimed at analyzing the serotypes of group B streptococcus (GBS) isolated from Korean infants with invasive disease and evaluating their association with disease manifestation. Methods: Data were retrospectively collected from invasive GBS infections at Gachon University Gil Medical Center from January 2006 to June 2012 and at Samsung Medical Center from April 2010 to November 2012. Serotypes were determined by slide agglutination test. Results: A total of 37 cases were identified, which included 22 full-term infants and 15 preterm infants. Fifteen cases (40.5%) were early-onset, 19 (51.4%) was late-onset, and three (8.1%) was very late-onset. Early-onset diseases among preterm infants were higher than those among full-term infants (60.0% [9/15] vs. 27.3% [6/22], P=0.17). The most common manifestation was bacteremia (70.3%), followed by meningitis and septic arthritis. Among 24 isolates retrievable for serotyping, serotype III (41.7%) was most common, followed by V (16.7%), Ia, Ib, and II (12.5%, respectively), and non-typeable (4.2%). Serotype III was more common in isolates from full-term infants (10/22) than from preterm infants (0/15), whereas serotype V was more common in isolates from preterm infants (4/15) than from full-term infants (0/22) (P=0.002). No penicillin-resistant strain was detected, and resistance to erythromycin and clindamycin were both 64.9%. Conclusions: GBS is an important pathogen in both preterm and full-term infants, and serotype distribution of GBS causing invasive diseases can differ between preterm and full-term infants. It is necessary to monitor the nationwide epidemiology of GBS diseases, including in preterm infants, in order to prepare preventive measures without underestimating early-onset diseases.

The Effect of Ecklonia stolonifera Extracts on Blood Flow and Serum Lipid Level in Ovariectomized Rats (곰피 추출물이 난소를 절제한 흰쥐의 혈액 유동성 및 혈중 지질함량에 미치는 영향)

  • Kim, Young-Kyoung;Jeon, Min-Hee;Park, Yong-Soo;Hwang, Hyun-Jung;Lee, Sang-Hyeon;Kim, Sung-Gu;Kim, Yuck-Young;Kim, Mi-Hyang
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1851-1858
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    • 2010
  • Estrogen deficiency in peri- and postmenopausal women results in variety of neurovegetative, psychic and somatic symptoms, and may contribute to severe diseases within the aged female population. Ecklonia stolonifera (ES) is an edible brown algae traditionally used in fishery towns in Far East Asia. This study was performed to investigate the effects of ES extracts on blood flow and serum lipid concentration in ovariectomized rats. Weight-matched female Sprague-Dawley strain rats were assigned to four groups. Three groups were surgically ovariectomized (OVX). The fourth group was sham operated. Rats were randomly assigned to the following groups: sham-operated rats (Sham), ovariectomized control rats (OVX-CON), ovariectomized rats supplemented with ES extract at 50 mg/kg bw/day (OVX-ES50) and ovariectomized rats supplemented with ES extract at 200 mg/kg bw/day (OVX-ES200). Serum total cholesterol and triglyceride contents decreased in the sham group compared to the OVX-CON group. Six weeks feeding of ES extract resulted in a significant (p<0.05) lowering of serum triglyceride and a lowering tendency of total cholesterol level. The level of Hdl-cholesterol in serum increased by supplementation of ES extracts at 50 mg/kg and 200 mg/kg bw/day (p<0.05). Blood passage time of the ES extracts-supplemented group was higher than the OVX-CON group. The ability of platelet aggregation of groups treated with ES extracts was less than that of the OVX-CON group. These results suggest that the beneficial effects of ES extract may be used to possibly improve on the metabolic syndrome of menopausal women.

A Study on the Making of Sweet Persimmon (Diospyros kaki, T) Wine (단감(Diospyros kaki, T) 와인 제조에 관한 연구)

  • Cho, Kye-Man;Lee, Jung-Bock;Kahng, Goon-Gjung;Seo, Weon-Taek
    • Korean Journal of Food Science and Technology
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    • v.38 no.6
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    • pp.785-792
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    • 2006
  • The characteristics of alcohol fermentation using sweet persimmon juice were studied in static fermentation in an effort to develop new types of functional wine. The yeast strain Saccharomyces cerevisiae KCCM 12650 was selected for use in the fermentation of sweet persimmon juice. Attempts were made to modify the sweet persimmon juice in order to find suitable conditions for alcohol fermentation. The modified sweet persimmon juice (pH 4.0) that was most suitable for alcohol fermentation contained $24^{\circ}Brix$ of sugar supplemented with sucrose as a carbon source and 0.5 g/L of $(NH_4)_2HPO_4$ as a nitrogen source. After 5 days of fermentation at $25^{\circ}C$, 12.8% of alcohol was produced from the modified juice and its pH was slightly decreased to 3.9. Browning of the wine was observed during storage due to the oxidation of phenolic compounds. The initial browning of 0.08% at $OD_{420}$ after fermentation increased to 0.40 during storage for 11 weeks at room temperature. The addition of $K_2S_2O_5$ was effective in delaying the browning of the wine. The browning of the wine decreased to 0.25 at $OD_{420}$ with the addition of 200 mg/L of $K_2S_2O_5$. The wine produced in this study contained some organic acids such as malic acid (6.82% g/L) and succinic acid (1.40 g/L), some minerals such as $K^+$ (947.8 mg/L) and $Mg^{2+}$ (36.4 mg/L), as well as soluble phenolics (779 mg/L of gallic acid equivalent). Schisandra fruit was added to the sweet persimmon juice during alcohol fermentation in order to improve the sour taste and flavor. The best sensory quality (taste, flavor, and color) was obtained by adding 0.5% schisandra fruit.

Activity and Survival of the Natural Bacteria under the Stressed Conditions Detected by Bioluminescent Phenotype (스트레스 하의 자연세균의 활성 및 생존의 발광표현형을 이용한 탐지)

  • Park, Kyoung-Je;Yoon, Hye-Young;Chun, Se-Jin;Lee, Ho-Sa;Lee, Dong-Hun;Jahng, Deokjin;Lee, Kyu-Ho
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.154-161
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    • 1998
  • To investigate whether the introduced genetic marker is useful to detect the survivalship and activity of the natural bacteria under the stressed conditions, one Gram-negative isolate, KP964 was transformed to the luminous phenotype by transferring luxAB gene. Under the starvation-stress this luminous bacterial culturability (determined by colony-forming-units [CFU] on agar plate) decreased rapidly below the detection limit by 37 days, while its total cell number (determined by AODC) remained almost the same as its initial inocular size. At that time period, the viable cell number was estimated to be 1400 times higher than its CFU number. The bioiuminescence (determined by relative light units [RLU]) produced under the same condition was also monitored and found to decrease more rapidly than the culturability by 5-fold. Under the other stresses, e.g., osmotic shocks, acid shock, and exposure to toxic chemicals, this bacterial strain did not show the reliable correlation between CFU and RLU. These results might not suggest the direct estimation of bioiuminescence from the stressed bacteria be an index of both the survivalship and its activity. However, when the stressed bacterial cells were incubated under the favorable condition by relieving from the existing stress, the potential bioiuminescence (the lag periods before the increase of bioiuminescence, the increase rates of bioiuminescence, and the maximal levels of bioiuminescence) was shown to be highly dependent upon the strengths of the stresses exposed to the bacterial cells. Therefore, analysis of the potential bioiuminescence from the stressed bacteria revealed good relationships with survival as well as activity.

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Biological Control of the Mushroom Fly, Lycoriella mali, Using Bacillus thuringiensis subsp. israelensis (Bacillus thuringiensis subsp. israelensis를 이용한 느타리 재배에서 버섯파리의 생물학적 방제)

  • Moon, Byung-Ju;Lee, Su-Hee;Lim, Eun-Kyung;Kim, Tae-Sung;Kim, Hyun-Ju;Song, Ju-Hee;Kim, Ik-Soo
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.50-55
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    • 2002
  • Biological control against mushroom fly, Lycoriella mali, was performed by using Bacillus thuringiensis subsp. israelensis Bti-D and Bti-U, isolated from dead mushroom fly in oyster mushroom houses. Control values of the bacterial strains Bti-D and Bti-U against L. mali in bottle culture of oyster mushroom were 74.4% and 64.2%, respectively, and the value in small tray culture were 75.8% and 56.8%, respectively. In the experiment to develop the mass, cheap media for Bti-D and Bti-U isolates, the Biji broth (bean curd residue, called Biji in Korean language) was selected as a culture medium for an inexpensive and mass cultivation by the measurement of optical density of the two bacteria grown in the different media tested. Insecticidal effect of the formulation contained different ingredients that were prepared by using the Bti-D strain cultured in the Biji broth was tested in tray and bottle culture of oyster mushroom. The WCS formulation that contained corn starch as bio-gel (86.4%) was more effective to control the mushroom fly than living cells (69.1%) in bottle culture of oyster mushroom. Moreover, insecticidal effect of the WCS formulation was improved when water of pH 8 was used for dilution of the formulation. Effect of the WCS formulation using water of pH 8 and chemicals, Zuron (dimillin) W.P. on the control of mushroom fly and the productivity of oyster mushroom was investigated in tray culture of oyster mushroom. The Zuron W.P. was more effective to control the mushroom fly than the WCS formulation. However, compared with no treatment, the productivity of the mushroom treated with the WCS formulation was improved than that of the mushroom with Zuron W.P.

Neural pathway innervating ductus Deferens of rats by pseudorabies virus and WGA-HRP (흰쥐에서 WGA-HRP와 pseudorabies virus를 이용한 정관의 신경로에 대한 연구)

  • Lee, Chang-Hyun;Chung, Ok-Bong;Ko, Byung-Moon;Lee, Bong-Hee;Kim, Soo-Myung;Kim, In-Shik;Yang, Hong-Hyun
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.11-24
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    • 2003
  • This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.

Thiobarbituric Acid Reactive Substances Levels in Brain Tissue of Aldh2 Knockout Mice Following Ethanol Exposure for 8 Weeks (Aldh2 knockout 마우스에서 8주간 에탄올 노출에 따른 뇌조직의 thiobarbituric acid reactive substances 농도)

  • Moon, Sun-In;Eom, Sang-Yong;Kim, Jung-Hyun;Yim, Dong-Hyuk;Kim, Hyong-Kyu;Kim, Yong-Dae;Kim, Heon
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1163-1167
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    • 2011
  • Excessive alcohol consumption causes various degenerative brain diseases including Alzheimer's disease and Parkinson's disease. Absorbed ethanol is metabolized to acetaldehyde and acetic acid by alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Acetaldehyde is well known as a toxicant through generation of reactive oxygen species (ROS). Therefore, ALDH2 activity may play important roles in the pathogenesis of alcohol-induced brain diseases. In this study, we demonstrated the effects of ALDH2 enzyme activity on lipid peroxidation in brain tissues and urine of mice exposed to ethanol for 8 weeks. Five male, 8-week old Aldh2 (+/+) and Aldh2 (-/-) mice (C57BL/6J strain) in each group were exposed to ethanol for 8 weeks (2 g/kg wt./day) using gavage, and those in the control group received 0.9% saline alone. Thiobarbituric acid reactive substances (TBARS) level, a marker for lipid peroxidation, was measured in whole brain tissue and urine by high performance liquid chromatography. As a result, chronic ethanol treatment did not show any statistical change on the TBARS level of brain tissue in both Aldh2 (+/+) mice and in Aldh2 (-/-) mice. However, following ethanol exposure for 8 weeks in Aldh2 (-/-) mice, the urinary TBARS levels were significantly increased to more than double compared to the pretreatment group. This result was not observed in Aldh2 (+/+) mice. These results suggest that although ALDH2 enzyme activity plays a role in the generation of ROS in the whole body, it does not seem to be important in the pathogenesis of alcohol induced degenerative brain diseases.