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Development of Simultaneous Analytical Method of Veterinary Antibiotics in Manure using Liquid Chromatography Coupled with Tandem Mass Spectrometry (LC-MS/MS를 이용한 퇴비 및 액비 중 항생제 동시 분석법 개발)

  • Chung, Hyung Suk;Lee, Young Jun;Lee, Han Sol;Rahman, Md. Musfiqur;Kabir, Md. Humayun;Park, Byung-Jun;Kim, Jang-Eok;Shim, Jae-Han
    • Korean Journal of Environmental Agriculture
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    • v.36 no.3
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    • pp.201-210
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    • 2017
  • BACKGROUND: The current study was to monitor of 9 veterinary antibiotics (ceftiofur, clopidol, florfenicol, sulfamethazine, sulfamethoxazole, sulfathiazole, tetracycline, tiamulin, and tylosin) in manure using liquid chromatography-tandem mass spectrometry (LC-MS/MS) in positive and negative electrospray ionization mode. METHODS AND RESULTS: Sample preparation was carried out using Mcllvaine buffer and citrate salts to adjust the pH of the sample followed by purification with dispersive solid phase extraction (d-SPE). Separation of analytes during LC-MS/MS analysis was conducted using an Eclipse Plus $C_{18}$ column and the mobile phase was in gradient mode with, 0.1% formic acid and 5 mM ammonium formate in methanol (A) and 0.1% formic acid and 5 mM ammonium formate in distilled water (B). The linearity of the matrix-matched calibrations of all tested antibiotics was good, with $R^2$ determination coefficients ${\geq}0.9920$. The limit of detection (LOD) and quantifications (LOQ) were $0.1-67.0{\mu}g/kg$ and $0.4-200.0{\mu}g/kg$, respectively. Analysis of 13 solid and liquid manure samples taken from the Republic of Korea revealed concentrations less than $0.7{\mu}g/kg$ for tiamulin, $1497.6{\mu}g/kg$ for sulfamethazine. CONCLUSION: To monitor 9 veterinary antibiotics from manure samples in 13 provincial areas throughout the Republic of Korea, an analytical method was developed. The developed method was fully validated and successfully applied for monitoring various veterinary antibiotics in manure samples.

Clinical Study on Hemolytic Uremic Syndrome in Children: Review of 23 Cases (소아 용혈성 요독 증후군 23예에 대한 임상적 고찰)

  • Oh Seungjin;Yook Jinwon;Kim Ji Hong;Kim Pyung-Kil
    • Childhood Kidney Diseases
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    • v.4 no.2
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    • pp.136-143
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    • 2000
  • Purpose : The hemolytic uremic syndrome (HUS) is characterized by microangiopathic hemolytic anemia thrombocytopenia, and acute renal failure. It is ole of tile most common cause of acute renal failure in children but few reports are available in Korea. Thus we investigated the 23 patients diagnosed as HUS during last 14 years. Method : We retrospectively investigated the etiologic factor, clinical manifestations laboratory findings, treatment modalities, and final outcomes of the patients. Then patients were divided into two groups according to outcome, md comparison was performed. Group A(8) comprised patients who progressed to end-stage renal disease or expired. Group B(15) comprised patients who completely recovered after dialysis treatment. Result The number of patients aged less than 4 years were 17; between 5 and 10 were 4 and more than 10 were 2. The gende ratio was M:F=2 : 1. The etiologic factors were as follows: acute gastroenteritis in 14 patients including 4 bloody diarrhea, upper respiratory tract infection in 7 patients, and 1 patient with herbal mediation. The overall mortality rate was 22$\%$: 2 patients died of US complications, 2 patients died of sepsis, and 1 patient died of pulmonary hemorrhage. Group A (Hb 4.8${\pm}$1.2 g/dL) showed lower value in hemoglobin than group B (Hb 6.3${\pm}$1.7 g/dL) during hospital stay (P< 0.05), And the time interval between tile disease onset and dialysis treatment was significantly longer in group A ($11.9{\pm}9.1\;days\;vs\;2.8{\pm}2.1\;days$) (P< 0.05). Conclusion : Overall mortality rate was 22$\%$. Low hemoglobin value and the prolonged time interval between the disease onset and dialysis treatment were related with poor prognosis. So early diagnosis and appropriate intensive care including dialysis treatment is essential to achieve better outcome in children.

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A comparative study of electric and manual toothbrushes on oral hygiene status in fixed orthodontic patients (고정식 교정 환자에서 전동치솔 효과에 관한 연구)

  • Park, Chang-Hun;Hwang, Hyeon-Shik;Lee, Ki-Heon;Hong, Suk_jin
    • The korean journal of orthodontics
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    • v.34 no.4 s.105
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    • pp.363-370
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    • 2004
  • Patients with fixed orthodontic appliances frequently have increased levels of plaque accumulation leading to the possibility of gingivitis or enamel decalcification. Although many methods may be helpful in reducing dental plaque formation, the optimal mechanical removal of plaque is the most important factor during orthodontic treatment. The purpose of this study was to evaluate the efficacy of an electric toothbrush (with a specially designed orthodontic brush head) compared to a manual toothbrush in controlling plaque and gingivitis for patients with fixed orthodontic appliances. Oral hygiene status was measured in thirty-four patients using a plaque index, a gingival index and a bleeding index, before and four weeks after the attachment of fixed orthodontic appliances. Patients were randomly divided into two groups: electric and manual toothbrush groups. Oral hygiene instruction was given according to the type of toothbrush used. The Braun Oral-B D9511 with Braun Oral-B Ortho OD 15-1 brush head was used as the electric toothbrush while the Butler G.U.M. 124 was given as the manual toothbrush. After four md eight weeks, oral hygiene status was measured again. Through a comparison between the electric and the manual toothbrush groups, the following results were obtained. 1. All oral hygiene indices showed an increasing tendency after four weeks of fixed orthodontic appliance. 2. All indices presented a decreasing tendency four and eight weeks after oral hygiene instruction. 3. In case of the gingival index and bleeding index, the decreasing tendency did not show a statistically significant difference between the electric and the manual toothbrush groups. 4. The decreasing tendency of plaque index presented a statistically significant difference between the two groups, showing that the electric toothbrush was more effective in terms of oral hygiene. These findings suggest that an electric toothbrush is useful to orthodontic patients with fixed appliances.

In vitro and In vivo Antibacterial Activity of a New Fluoroquinolone Containing C7-bicyclic Structure (C7-이환체 구조를 갖는 새로운 플루오르퀴놀론계 항생제의 in vitro와 in vivo 항균작용)

  • Han, Seung-Hui;Choe, Mun-Jeong;Kim, Ji-Yeon;Kim, Byeong-O;Sim, Jeom-Sun;Gang, Jin-Seok;Son, Ho-Jeong;Lee, Jae-Uk;Yu, Yeong-Hyo;Park, Myeong-Hwan
    • YAKHAK HOEJI
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    • v.40 no.4
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    • pp.428-437
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    • 1996
  • The in vitro and in vivo antibacterial activities of a new fluoroquinolone, DWP20364(1-cyclopropyl-5-amino-6,8-difluoro-7-(2,7-diazabicyclo[3.3.0]oto-4-ene-7-yl)-1 ,4-di-hydro-4-oxoquinoline-3-carboxylic acid) were evaluated in comparison with those of ciprofloxacin(CPFX), sparfloxacin(SPFX) and ofloxacin(OFLX). DWP20364 was more potent than CPFX and OFLX against Staphylococcus spp., Streptococcus spp. and Enterococcus faecium MD8b and it was similarly or slightly less active than CPFX against Escherichia spp. and Pseudomonas spp.. For MRSA and OFLX resistant strains (Staphylococcus spp.(14),Enterococcus spp.(4), Acinetobacter spp.(2), Pseudomonas spp.(9), Klebsiella spp.(2) and Serratia spp.(6)),DWP20364(MICs for 90% of strains,0.025 and 12.5${\mu}$g/ml, respectively) was 4 to 32 folds more potent than SPFX and CPFX. The activity of DWP20364 decreased moderately in the presence of 5mM $Mg^{2+}$. However, various pHs and the concentrations of various serum had no effect on the activity of DWP20364. DWP20364 possessed a bacteriocidal effect at the 1MIC against gram positive and gram negative strains. The protective effect of DWP20364 against systemic infections in mice caused by S. aureus Smith or S. aureus L2379 was superior to that of CPFX and SPFX but it was less active than that of CPFX against infection by P. aeruginosa E-2.

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Changes in the Profiles of Serum LH, Testosterone, Estrogen and IFG-I during Sexual Development in Male Korean Native Chickens (한국재래 수탉에서 부화 후 혈청내 LH, Testosterone, Estrogen과 IGF-I 농도의 변화)

  • Tae H. J.;Jang B. G.;Choi C. H.;Park Y. J.;Yang H. H.;Kim I. S.
    • Korean Journal of Poultry Science
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    • v.32 no.2
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    • pp.135-141
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    • 2005
  • Changes in serum estradiol, insulin-like growth factor-1, leuteinizing and testosterone levels, and leuteinizing hormone-stimulated testosterone production per testis in vitro from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The changes in the profiles of the levels in the incubation medium of luteinizing hormone-stimulated (100 ng/mL) testosterone secretion per testis in vitro, and the serum LH, testosterone, estradiol, and insulin-like growth factor-I were determined by radioimmunoassay. Serum estradiol levels were not significantly different at week 4 compared to that of 8, 12, 16, 21, 32, :md 44. Significant decreases were observed at weeks 52 and 64. Serum leuteinizing hormone concentrations were not significantly different from 1 week to 12 weeks, increased gradually up to 32 weeks of age, and declined significantly thereafter; the highest value was at 32 weeks, and the lowest value was detected at 2 weeks of age. Serum insulin-like growth factor-I concentrations increased significantly from 1 week to 16 weeks, remained low and unchanged with advancing age. Serum testosterone concentrations were not significantly different at week 1 compared weeks 2, 4, 6, and 8. Significant increases were observed from 10 weeks to 32 weeks of age. Values at weeks 24, 28 and 32 and at weeks 32, 44, 52, and 64 were not significantly different. The highest value was at weeks 28 and the lowest value was detected at weeks 1 week. LH-stimulated testosterone production per testis in vitro increased gradually with age from 1 to 32 weeks and decreased significantly from 44 weeks to 64 weeks of age.

Bacterial Community Dynamics during Swine In vitro Fermentation Using Starch as a Substrate with Different Feed Additives for Odor Reduction

  • Alam, Md.J.;Jeong, C.D.;Mamuad, L.L.;Sung, H.G.;Kim, D.W.;Cho, S.B.;Lee, K.;Jeon, C.O.;Lee, Sang-S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.690-700
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    • 2012
  • The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen ($NH_3$-N) and $H_2S$ gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, $NH_3$-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as $NO_2$, $NO_3$ and $SO_4$ concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that $NH_3$-N, $H_2S$ and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds.