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Study on Low Temperature Tolerant Methane-Producing Bacteria for the Treatment of Agricultural and Livestock Wastes;III. Isolation of Low Temperature Tolerant Methanogens (농축산(農畜産) 폐기물(廢棄物) 처리(處理)를 위(爲)한 저온내성(低溫耐性) 메탄 생성균(生成菌)의 특성(特性)에 관(關)한 연구(硏究);III. 저온내성(低溫耐性) Methanogens의 분리(分離))

  • Kim, Kwang-Yong;Kim, Jai-Joung;Daniels, Lacy
    • Korean Journal of Environmental Agriculture
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    • v.15 no.3
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    • pp.362-371
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    • 1996
  • This study was conducted to investigate the biochemical properties of isolated bacteria, low temperature tolerant methanogens which were selected for use as inoculum for anaerobic fermentation of agricultural and livestock wasted at low temperature. The results, obtained were summarized as follows: Low temperature tolerant methanogens were isolated from the samples which showed the high methanogenesis rate by enrichment culture at low temperature in methanol medium. These methanogens, Methanobacterium M-251 and Methanobacterium M-253 were isolated from swampy sediment at latitude $56.9^{\circ}$, Methanosarcina mazei M-372 from lake sediment IV at latitude $55.0^{\circ}$ N, and Methanobacterium formicicum M-375 from tidal land soil at latitude $37.0^{\circ}N$, respectively. The isolated anaerobic bacteria could not use sugars as carbon sources. The optimum pH value for the growth of M-251 and M-375 was 6.8, but those for M-253 and M-372 6.5 and 7.0, respectively. The minimum growth temperature of isolated, M-251 and M-253 were $8^{\circ}C$ and the optimum temperature $30^{\circ}C$, while the minimum of M-392 and M-395 were $13^{\circ}C$ and the optimum $37^{\circ}C$. The growth rate of isolates at $17.5^{\circ}C$ were lower by 32-50% than that of $30^{\circ}C$. The isolated Methanobacterium strains such as M-251, M-253, and M-375 have lower cell yield, 0.38-1.21g/1M $CH_4$ than 1.14-1.51g/1M $CH_4$ of Methanosarcina mazei M-372.

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Development of a Duplex RT-PCR Assay for the Simultaneous Detection and Discrimination of Avirulent and Virulent Newcastle Disease Virus (NDV) (뉴캣슬병 바이러스 검출 및 병원성 감별을 위한 Duplex RT-PCR법 개발)

  • Kim, Ji-Ye;Lee, Hyun-Jeong;Jang, Il;Lee, Hee-Soo;Yoon, Seung-Jun;Park, Ji-Sung;Seol, Jae-Goo;Kim, Seung-Han;Hong, Ji-Mu;Wang, Zillian;Liu, Hualei;Choi, Kang-Seuk
    • Korean Journal of Poultry Science
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    • v.44 no.2
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    • pp.93-102
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    • 2017
  • A duplex RT-PCR (dRT-PCR) assay was developed for the simultaneous detection and discrimination of non-virulent and virulent Newcastle disease virus (NDV) in a single PCR tube. Primers targeting the large polymerase protein (L) gene and the fusion protein (F) gene of NDV were designed to detect all NDVs (by common type PCR primers) and virulent NDVs (by pathotype PCR primers), respectively and evaluated experimentally with reference NDV strains and other poultry viral pathogens. PCR products of the expected size of 386 bp were amplified from all NDV samples whereas PCR products of the expected size of 229 bp were amplified from virulent NDV samples alone. Cross reaction was not observed with other avian viral pathogens. The detection limit of NDV by the dRT-PCR was estimated to be $10^3$ 50% egg infectious dose/0.1 mL. In the dRT-PCR using field isolates of NDV, the pathotype PCR primers detected specifically all of virulent field isolates of NDV from Malaysia, Pakistan and China whereas common type PCR primers detected 94.4% (51/54) of field isolates of NDV from China. Three Chinese NDV isolates with false negative result were non-virulent viruses. Our results indicate that the dRT-PCR might provide a rapid and simple tool for rapid simultaneous detection and discrimination of non-virulent and virulent NDVs. Therefore the developed dRT-PCR assay provides a powerful novel means for the rapid diagnosis of Newcastle disease.

Use of Antimicrobial Agents for the Treatment of Inpatients in Chonbuk National University Hospital (전북대학교병원 입원환자에서의 항생제 사용 실태)

  • Song, Jae Ho;Kim, Jung Soo
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.225-232
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    • 2000
  • Purpose : The use antimicrobial agents is one of the important strategies for the treatment and prophylaxis of microbial infections. But injudicious abuse and misuse of antimicrobial agents is problem to add an extra weight on medical fee, increase of resistant bacteria and side effects according to the antibiotic use. This study was performed to establish the pertinent use of antimicrobial agent in Chonbuk National University Hospital(CNUH). Characteristics of antibiotics use was analysis by reviewing the medical records of patients admitted to CNUH during the period of May 1998. Methods : One thousand eight hundred and thirty three patients were enrolled in this study(medical division 1,014 cases, surgical division 819 cases). Medical records were retrospectively reviewed to classify the rate of antibiotics use, name of antibiotics used, appropriateness of antibiotics use. Results : The overall rate of antibiotic usage in CNUH was 67.2%(1,231/1,833), showing higher rate in surgical division(89.6%) compare to that of medical division(49.0%). Among 1,231 patients to whom antimicrobial agents were given, only 125(10.2%) were treated with single antimicrobial agents. 311(25.3%) were treated with two antimicrobial agents, and 795(64.5%) patients received 3 or more antibiotics. ${\beta}$-lactams(56.4%) were most frequently used followed by aminoglycosides(35.3%), the others(4.9%) and quinolons(3.4%). Amoxicillin-clavulanate was the mostly commonly used antibiotics followed by amoxicillin and unasyn. Prophylactic use of antibiotics was carried in seven hundred six patients(57.4%), mostly in surgical division, which can be considered somewhat inappropriate in the initiation time and duration of antibiotic use. Conclusion : Importance of monotherapy and appropriate prophylactic antibiotic use should be emphasized. Strategies of antibiotics use, such as restriction of drug use, continuous monitoring system, flow sheet system should be considered to reduce antibiotics use and establish the appropriate use of antibiotics as well as inhibiting the occurrence of resistant strains.

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Salmonellosis in Children in Daejeon, Korea, 1994~1999 (대전지역 살모넬라증 환아에 대한 임상적 연구(1994~1999년))

  • Chun, Jung-Soo;Lee, Kyung-Yil;Lee, Hyung-Shin;Kim, Sang-Yong;Han, Ji-Whan;Her, Jae-Kyun;Whang, Tyung-Tai
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.211-217
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    • 2000
  • Purpose : Salmonellosis including typhoid fever is still prevalent in Korea. Recently it has been reported that the incidence of salmonella gastroenteritis is increasing with a reduction of typhoid fever. We studied the clinical and laboratory features of salmonellosis. Methods : We evaluated the clinical records of 83 patients with salmonellosis who had been confirmed by stool culture from 1994 to 1999. Results : Annual incidence of salmonellosis showed an increasing tendency during recent 4 years. Seasonally, summer(45.8%) was the most prevalent, followed by fall(32.5%). Male to female ratio was 1.4 : 1. In age distribution, 64 cases(77.1%) were under 5 year-old, and 18 cases(21.7%) were younger than 1 year-old. Clinical features included diarrhea(96.4%), fever(91.6%), vomiting(49.4%), bloody stool(42.1%), abdominal pain(40.1%) and tenesmus(12.0%). In serogroups, there was no group A and group B, group C, group D(including 2 cases of S.typhi) and group E were in 41.0%(34 cases), 3.6%(3 csaes), 51.8%(43cases) and 3.6%(3 cases), respectively. In Widal test, 5 cases(13.5%) and 1 case(2.7%) of group B, C and E(total 37 cases) were observed the O titer above 1 : 80 and 1 : 320, respectively. However, in 36 cases of group D, 19 cases(52.7%) and 9 cases(25.0%) were above the O titer 1 : 80 and 1 : 320, respectively. Antibiotics resistant rates to ampicillin, trimethoprim/sulfamethoxazole and chloramphenicol were 23.2%, 10.1% and 51.4%, respectively. Conclusion : Salmonellosis has become a common cause of gastroenteritis in children, especially under the pre-school age. Isolation of salmonella is neccessary for accurate diagnosis of gastroenteritis and typhoid fever. A careful attention of the use of antibiotics is needed to reduce the muilti-drug resistant strains.

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Genotyping and Clinical study of Mycoplasma pneumoniae During 1996-1997 in Korea (1996년과 1997년 소아에서 유행한 Mycoplasma pneumoniae의 Genotype에 따른 임상적 연구)

  • Lim, Hong Hee;Ahn, Byung Moon;Kim, Eun Ryoung;Choi, Sug Ho;Moon, Young Ho;Kim, Il Soo
    • Pediatric Infection and Vaccine
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    • v.5 no.1
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    • pp.79-87
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    • 1998
  • Purpose : The P1 protein of Mycoplasma pneumoniae mediates the attachment of the pathogen to its host cell and elicits a strong humoral immune response during infection with this organism. Mycoplasma pneumoniae strains can be classified into two groups(I and II) by PCR method of P1 cytadhesin gene. In this study, we evaluated the prevalence, epidemiological and clinical characteristics of each group. Methods : From 155 patients with Mycoplasma pneumoniae, who admitted to the Department of Pediatrics, Sung-Ae and Kwangmyung Sung-Ae Hospital between November 1996 and October 1997, we collected their throat swabs or nasopharyngeal aspirates for DNA extraction and serum for indirect hemagglutination test of Mycoplasma pneumoniae. The group specific PCR amplification were performed using specific oligonucleotide primers designed for P1 gene genotyping. Results : Group I(137 patients, 88.4%) occurred frequently than group II(18 patients, 11.6%). In both group, the most prevalent season was winter in 1996(Nov. to Dec.) and fall in 1997(Aug. to Oct.) The prevalent age was four to six years old. The number of male was more than female in both group; Group 1(1.2:1), Goup 2(1.6:1). No significant relationship were found between two groups in duration of fever and hospital days(P>0.05). The rate of high antibody titers(>1:5120) was lower in group I(6/137, 4.4%) than group II(2/18, 11.1%). Conclusion : Group I was much more prevalent than group II during 1996~1997 in Korea. There was no difference between two groups in epidemiological and clinical parameters except the rate of high antibody titers. Further follow-up survey will be needed for the epidemiologic and clinical studies of Mycoplasma pneumoniae in Korea.

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Rotavirus P and G Genotypes Circulating in Kyungsangnamdo, Korea, during 2000~2001 (2000~2001년 경상남도에서 유행한 로타바이러스 유전자형)

  • So, Kyeung Jin;Lee, Mi Hyun;Ma, Sang Hyeok;Kim, Byung Chyeol;Yang, Jai Myung
    • Pediatric Infection and Vaccine
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    • v.11 no.1
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    • pp.59-72
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    • 2004
  • Purpose : Rotaviruses are the major cause of gastroenteritis in infants and young children worldwide. It is important to get the epidemiologic data of rotavirus genotype for the application of rotavirus vaccine. So we tried to investigate the distribution of rotavirus genotypes with RT-PCR. Methods : A total of 120 rotavirus latex agglutinin test positive stool samples were collected continually from 120 children from Sep. 2000 to Apr. 2001. Rotavirus P(VP4), G (VP7) genotypes were determined by RT-PCR. Results : The genotype was identified in 116 stool samples of total 120 samples(96%). The incidence of G genotype was as follow; G1 17(14.2%), G2 74(61.7%), G4 1(0.8%), G9 1(0.8%). There were four cases of multiple genotypes; G1/G2, G1/G4, G1/G9, G8/G9 and genotype of G3, G8 were not found. Twenty three(19.2%) samples were nontypeable. The incidence of P was as follow; P[4] 77(64.2%), P[6] 22(18.3%), P4/P6 12(10%), P[4]/P[8] 1(0.8%) p[8] 1(0.8%). Seven(5.9%) samples were nontypeable. Conclusion : Various combinations of G and P genotypes were observed. Most rotavirus strains were P[4]G2 62(51.74%), followed by P[6]G2 7(5.8%), and P[6]G1 7(5.8%), P[4/P[6] G1 4(3%), P[4]/P[6]G2 4(3%), P[4]G1 3(2.5%), P[8]G2 1(0.8%), P[4]G4 1(0.8%) in Kyoungsangnamdo, Korea during 2000~2001.

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Seasonal Distribution of T Serotyping and emm Genotyping of Group A Streptococci Obtained from Children with Streptococcal Infections in Masan, Korea, 2003~2004 (2003~2004년 경남 마산 지역에서 분리된 A군 연쇄구균의 T 혈청형과 emm 유전자형의 계절별 분포)

  • Jeon, Ho-Sang;Park, Hwa-Jin;Lee, Hee-Joo;Ma, Sang-Hyuk;Cha, Sung-Ho
    • Pediatric Infection and Vaccine
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    • v.12 no.1
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    • pp.52-60
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    • 2005
  • Purpose : The aim of this study is to know seasonal distribution of group A streptococci obtained from one center using emm genotyping and T serotyping in Masan from 2003 through 2004. Methods : Among children who visited the Changwon Fatima Hospital at Masan, Korea from June 2003 through February 2004, 100 patients who had clinical findings of acute pharyngitis, scarlet fever, and cellulitis were confirmed as GAS by culture, and were enrolled in our study. All obtained GAS were sent to the WHO Collaborative Center for Reference and Research on Streptococci, University of Minnesota, Minneapolis for T serotyping and emm genotyping. We classified these results again according to seasonal and disease's entities. Results : 19 different T serotypes was typed. T4(27.5%), T1(17.6%), T6(13.7%), and T12(13.7%) serotypes were relatively common in summer, while T4(28.3%), T12(15.2%), and T12/B3264(8.7%) were common in winter. T4 and T12 were persistent all year around. Distribution of T serotypes in 89 patients with pharyngotonsillitis were T4(26.7%), T12(14.0%), T1(12.8%), and T6(11.6%) in order of frequency. 15 different emm genotypes was typed. The number of emm 1, emm 6, emm 9, and emm 44 genotypes decreased or disappeared in winter, and the number of emm 3, emm 12, and emm 89 genotypes increased or reappeared in winter. Conclusion : Because T serotyping and emm genotyping are useful tools for evaluating epidemiology and pathogenesis of group A streptococci, we should monitor these strains every year, and should serotype and genotype GAS obtained from the invasive streptococcal infections.

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Antimicrobial resistance patterns of Listeria species and Staphylococcus aureus isolated from poultry carcasses in Korea (계육에서 분리한 Listeria species 와 Staphylococcus aureus의 항생제 내성패턴)

  • Hur, Jin;Kim, Jun Man;Kwon, Nam Hoon;Park, Kun Taek;Lim, Ji Youn;Jung, Woo Kyoung;Hong, Soon Keun;Park, Yong Ho
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.217-224
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    • 2004
  • This study was carried out to investigate the antibiotic resistance pattern of Listeria spp. and Staphylococcus aureus. A total of 17 (14.8%) L. monocytogenes, 13 (11.3%) L. innocua, 7 (7%) L. welshimeri, and 83 (72.2%) S. aureus were isolated from commercial poultry carcasses in Seoul and Kyonggi province during the period between 2001 and 2003. Antibiotic susceptibility test of all Listeria strains isolated was performed by the disk agar diffusion method. Antibiotics used in the study were as follows; Amikacin (An), Ampicillin (Am), Cephalothin (Cf), Chloramphenicol (C), Ciprofloxacin (Cip), Erythromycin (E), Gentamicin (Gm), Imipenem (Ipm), Kanamycin (K), Minocycline (Mi), Neomycin (N), Norfloxacin (Nor), Ofloxacin (Ofx), Penicillin (P), Streptomycin (S), Tetracycline (Te), Tobramycin (Nn), Trimethoprim (Tmp), Trimethoprim/Sulfamethoxazloe (Sxt), and Vancomycin (Va). The antibiotic resistance pattern of S. aureus isolates was performed by the disk agar diffusion method. For the latter program, antibiotics used to the study were as follows; Cf, C, Cip, Clindamycin (Cc), E, Gm, Ipm, Nafcillin (Nf), Oxacillin (Ox), P, Te, Sxt, and Va. Of the 17 L. monocytogenes isolates, 94.1% were resistant to Te, 88.2% to Mi, 11.8% to Nor, 11.8% to S, 5.9% to Cip, and 5.9% to C. Of 13 L. innocua, 53.8% were resistant to Te, 23.1% to Mi, 23.1% to S, 7.7% to Cip, and 7.7% to Nor. Of 7 L. welshimeri, 57.1% were resistant to Te, and 14.3% to Am. Of 83 S. aureus, 100% were resistant to Te, 86.7% to Gm, 34.9% to P, 15.7% to Cip, 12% to Cc, 9.6% to E. The multiple antibiotic resistance patterns of L. monocytogenes isolates were observed in Te Mi Cip (5.9%), Te Mi Nor (5.9%), Te Mi (76.5%), and Te Nor (5.9%). Multiple antibiotic resistance was also found in L. innocua isolates. Resistant to Te Mi S Cip Nor was 7.7%, Te Mi S (7.7%), Te Mi (7.7%), and was 7.7% to Te S. Antibiotic resistance patterns for S. aureus isolats were demonstrated to Te Gm P Cip Cc E (6.0%), Te Gm Cip Cc E (3.6%), Te Gm P Cc (1.2%), Te Gm P (15.6%), Te Gm Cip (2.4%), Te P Cip (2.4%), Te Gm Cc (1.2%), Te Gm (56.6%), Te P (9.6%), and to Te Cip (1.2%). The results of this study suggest a high incidence of Lsteria spp. and S. aureus on poultry carcasses. The contaminated poultry carcasses may be a potential vehicle for foodborne infections due to multiple antimicrobial resistant organisms.

Characterization and Volatile Flavor Components in Glutinous Rice Wines Prepared with Different Yeasts of Nuruks (누룩에서 분리한 효모를 이용한 찹쌀발효주의 이화학적 특성 및 휘발성 향기성분)

  • Kim, Hye-Ryun;Kwon, Young-Hee;Jo, Sung-Jin;Kim, Jae-Ho;Ahn, Byung-Hak
    • Korean Journal of Food Science and Technology
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    • v.41 no.3
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    • pp.296-301
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    • 2009
  • In order to investigate the effect of different yeasts (La Parisienne (LP), Y18-2, Y54-3, Y90-2, Y90-9 and Y272-7) from nuruks on the quality of Glutinous rice wines, physicochemical properties and volatile flavor components were evaluated. Glutinous rice wines prepared with different yeasts were analyzed for ethanol, pH, total acid, amino acid, soluble solid, coloring degree, UV absorbance, reducing sugar, organic acid, free sugar and volatile compounds. After fermentation for 17 days, the ethanol contents ranged from 13.40 to 14.50%, while the total acid levels were from 0.33 to 0.44%. The amino acid contents in six samples ranged from 0.13 to 0.18%, while soluble solid contents ranged from 12.1 to $14.7^{\circ}Bx$. The glutinous rice wine prepared with LP showed the highest level of coloring degree, soluble solid and reducing sugar among six samples. Organic acid contents of the glutinous rice wine prepared with LP had the highest levels of lactic acid and acetic acid, while the glutinous rice wine prepared with Y90-9 had the highest level of succinic acid. In all glutinous rice wines tested, the most abundant free sugars were glucose followed by maltose. Volatile flavor components in the glutinous rice wines were identified by using GC-MSD. Nineteen esters, ten alcohols, eight acids, one aldehyde and one miscellaneous compound were identified in the glutinous rice wines. Using relative peak area, it was found that other than ethyl alcohol, hexadecanoic acid ethyl ester was the major component, predominantly found in the range of 2.73-10.41%. Phenylethyl alcohol, isoamyl alcohol, ethyl oleate, ethyl linoleate and tetradecanoic acid ethyl ester were some of the major volatile components present through the fermentation, respectively. Overall, it was shown that different yeast strains from nuruks greatly affected chemical and volatile characteristics of the glutinous rice wines.

Isolation and Characterization of Bacillus Strain as a Potential Biocontrol Agent (환경친화적 미생물농약으로서의 잠재성을 가진 세균의 분리 및 특성)

  • Lee, Ye-Ram;Lee, Sang-Mee;Jang, Eun-Young;Hong, Chang-Oh;Kim, Keun-Ki;Park, Hyean-Cheal;g Lee, Sang-Mon;Kim, Young-Gyun;Son, Hong-Joo
    • Journal of Life Science
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    • v.25 no.12
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    • pp.1408-1414
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    • 2015
  • In this study, to retain a stable bacterial inoculant, Bacillus strains showing antifungal activity were screened. The improved production, antifungal mechanism, and stability of the antifungal metabolite by a selected strain, AF4, a potent antagonist against phytopathogenic Botrytis cinerea, were also investigated. The AF4 strain was isolated from rhizospheric soil of hot pepper and identified as Bacillus subtilis by phenotypic characters and 16S rRNA gene analysis. Strain AF4 did not produce antifungal activity in the absence of a nitrogen source and produced antifungal activity at a broad range of temperatures (25-40℃) and pH (7-10). Optimal carbon and nitrogen sources for the production of antifungal activity were glycerol and casein, respectively. Under improved conditions, the maximum antifungal activity was 140±3 AU/ml, which was higher than in the basal medium. Photomicrographs of strain AF4-treated B. cinerea showed morphological abnormalities of fungal mycelia, demonstrating the role of the antifungal metabolite. The B. subtilis AF4 culture exhibited broad antifungal activity against several phytopathogenic fungi. The antifungal activity was heat-, pH-, solvent-, and protease-stable, indicating its nonproteinous nature. These results suggest that B. subtilis AF4 is a potential candidate for the control of phytopathogenic fungi-derived plant diseases.