• Title/Summary/Keyword: anaerobic culture

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INDIRECT IMMUNOFLUORESCENCE FOR THE IDENTIFICATION OF ACTINOMYCES SPECIES IN PATIENTS WITH PERIAPICAL LESION (치근단 병변에서 Actinomyces 종의 검정을 위한 간접 면역형광법적 연구)

  • Chang, Won-Jung;Yoon, Soo-Han;Kwon, O-Yang
    • Restorative Dentistry and Endodontics
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    • v.21 no.1
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    • pp.121-135
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    • 1996
  • Actinomyces are Gram-positive, non-acid-fast, anaerobic or microaerophilic filamentous bacteria. These organisms are frequently detected from infected root canals and periapical lesion. The purpose of this study was to use indirect immunofluorescence to determine the prescence of select Actinomyces species in a survey of teeth associated with periapical lesion, to clarify the relationship between clinical symptoms of periapical lesions and the Actinomyces species and to study on the cross reaction among Actinomyces. Actinomyces israelii serotype I (ATCC 12102), Actinomyces israelii serotype II (ATCC 29322), Actinomyces viscosus serotype II (ATCC 19246), Actinomyces naslundii serotype I (ATCC 12104) were cultured in anaerobic condition. Rabbit antisera were prepared by intravenous injection of formalized whole cells. Indirect immunofluorescence method was used to achieve the purpose. The following results were obtained. 1. There was a relationship between Actinomyces and periapical disease. 2. A. israelii serotype I, II were frequently identified with Indirect Immunofluorescence and most often assosiated with periapical disease. In culture finding, there was no significant difference between each group. 3. Indirect Immunofluoresence is both more sensitive and more rapid than culture for identification of Actinomyces species in patients with periapical lesion. 4. A. israelii serotype I, II was highly isolated in infected root canals with local swelling, A. naslundii serotype I was highly isolated in those with foul odor, and A. israelii serotype I was found in higher frequncy in those with exudate than other bacteria. 5. In the Indirect Immunofluorescence (1 : 320), A positive cross reaction was obtained between A. israelii serotype I and A. israelii serotype II, also, A. viscosus serotype II and A. naslundii serotype I. There was no cross reaction between A. israelii serotype I, II and A. viscosus serotype II, A. naslundii serotype I.

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Isolation and Identification of a Lactic Acid Bacterial Strain KJ-108 and Its Capability for Deodorizing Malodorous Gases Under Anaerobic Culture Conditions

  • KIM, JEONG-DONG;JUNG-HOON YOON;YONG-HA PARK;DAE-WEON LEE;KYOU-SEUNG LEE;CHANG-HYUN CHOI;WON-YEOP PARK;KOOK-HEE KANG
    • Journal of Microbiology and Biotechnology
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    • v.13 no.2
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    • pp.207-216
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    • 2003
  • A number of different sources, such as composts, leachates, and pig feces samples, were collected from different pig farms in Korea, and several microorganisms were screened for their ability to deodorize the malodorous gases. Consequently, a novel malodorous gases-deodorizing bacterial strain, KJ-108. was isolated, because it was highly abundant in nitrate-supplemented minimal medium ($MM-NO_3^-$) under anaerobic culture conditions. Airtight crimp-sealed serum bottles containing $MM-NO_3^-$ , medium were inoculated with KJ-108. Nitrate concentration was decreased rapidly after 20 h of incubation, and incubation was carried out until nitrite production reached almost zero. Taxonomic identification, including 16S rDNA base sequencing and phylogenetic analysis, indicated that the isolate had $100\%$ homology in its 165 rDNA base sequence with Lactobacillus pentosus. Among the volatile fatty acids, acetic acid contained in large amounts in fresh piggery slurry was decreased by about $40\%$ after 50 h incubation with strain KJ-108. n-Butyric acid, n-valeric acid, and isovaleric acid were gradually decreased, and isobutyric acid and capronic acid were dramatically eliminated at theinitial period with the treatment. Moreover, NH, removal efficiency reached a maximum of $98.5\%$ after 50 h of incubation, but the concentration of $H_2S$ was not changed.

Identification and Antibiotic Susceptibility of the Bacteria from Non-odontogenic Infectious Lesions

  • Kim, Yong Min;Kim, Jae-Jin;Kim, Mija;Park, Soon-Nang;Kim, Hwa-Sook;Kook, Joong-Ki;Kim, Hak Kyun
    • International Journal of Oral Biology
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    • v.39 no.2
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    • pp.87-95
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    • 2014
  • The purpose of this study was to isolate and identify bacteria from the 4 patients with non-odontogenic infectious lesions (mucormycosis, chronic inflammation from wound infection, and two actinomycosis) and determine their antimicrobial susceptibility against eight antibiotics. Bacterial culture was performed under three culture conditions (anaerobic, $CO_2$, and aerobic incubator). The bacterial strains were identified by 16S rRNA gene (16S rDNA) sequence comparison analysis method. For investigating the antimicrobial susceptibility of the bacteria against eight antibiotics, penicillin G, amoxicillin, tetracycline, cefuroxime, erythromycin, clindamycin, vancomycin, and Augmentin$^{(R)}$ (amoxicillin + clavulanic acid), minimum inhibitory concentration (MIC) measurement was performed using broth microdilution assay. Nosocomial pathogens such as Enterococcus faecalis, Klebsiella pneumoniae, Bacillus subtilis, and Neisseria flavescens were isolated from mucormycosis. Veillonella parvula, Enterobacter hormaechei, and Acinetobacter calcoaceticus were isolated from chronic inflammatory lesion. Actinomyces massiliensis was isolated from actinomycosis in parotid gland. Capnocytophaga ochracea was isolated from actinomycosis in buccal region in anaerobic condition. There was no susceptible antibiotic to all bacteria in mucormycosis. Tetracycline was susceptible to all bacteria in chronic inflammation. C. ochracea was resistant to vancomycin and penicillin G; and other antibiotics showed susceptibility to all bacteria in actinomycosis. The results indicated that the combined treatment of two or more antibiotics is better than single antibiotic treatment in mucormycosis, and penicillin is the first recommended antibiotic to treat actinomycosis.

Composition and Diversity of Gut Bacteria Associated with the Eri Silk Moth, Samia ricini, (Lepidoptera: Saturniidae) as Revealed by Culture-Dependent and Metagenomics Analysis

  • MsangoSoko, Kondwani;Gandotra, Sakshi;Chandel, Rahul Kumar;Sharma, Kirti;Ramakrishinan, Balasubramanian;Subramanian, Sabtharishi
    • Journal of Microbiology and Biotechnology
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    • v.30 no.9
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    • pp.1367-1378
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    • 2020
  • The polyphagous eri silk moth, Samia ricini, is associated with various symbiotic gut bacteria believed to provide several benefits to the host. The larvae of S. ricini were subjected to isolation of gut bacteria using culture-dependent 16S rRNA generic characterization, metagenomics analysis and qualitative enzymatic assays. Sixty culturable aerobic gut bacterial isolates comprising Firmicutes (54%) and Proteobacteria (46%); and twelve culturable facultative anaerobic bacteria comprising Proteobacteria (92%) and Firmicutes (8%) were identified inhabiting the gut of S. ricini. The results of metagenomics analysis revealed the presence of a diverse community of both culturable and un-culturable gut bacteria belonging to Proteobacteria (60%) and Firmicutes (20%) associated with seven orders. An analysis of the results of culturable isolation indicates that these bacterial isolates inhabited all the three compartments of the gut. Investigation on persistence of bacteria coupled with metagenomics analysis of the fifth instar suggested that bacteria persist in the gut across the different instar stages. In addition, enzymatic assays indicated that 48 and 75% of culturable aerobic, and 75% of anaerobic gut bacterial isolates had cellulolytic, lipolytic and nitrate reductase activities, thus suggesting that they may be involved in food digestion and nutritional provision to the host. These bacterial isolates may be good sources for profiling novel genes and biomolecules for biotechnological application.

Most Probable Number 방법을 이용하여 측정한 중랑천 하상토양의 혐기성 세균의 수와 수질과의 상관 관계

  • Park, Doo-Hyun;Kim, Byung-Hong;Lim, Si-Keun;Choi, Young-Hyo
    • Microbiology and Biotechnology Letters
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    • v.24 no.3
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    • pp.364-370
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    • 1996
  • Sediments collected from the Jungnang-cheon and its tributaries were used to enumerate anaerobic bacteria by most probable number (MPN) methods. A simple method was developed to detect ferrous ion in the culture fluid in order to count the number of iron ion reducers, and sulfate-reducing bacteria (SRB) and methanogens were detected by the presence of FeS precipitate in the culture or methane in the head space, respectively. The numbers of iron reducer was in the range of 10$^{7}$ - 10$^{8}$ /g in the sediment of the stream containing higher organic content than the tributaries. The sediments of tributaries were analyzed to contain iron reducers less than 10$^{7}$ cells/g. With one exception the numbers of SRB and methanogens were less than 10$^{3}$ cells/g in the sediment. From these results it is concluded that organics in the sediment support the growth of iron reducers, which out-compete SRB and methanogens.

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Kinetic Study of pH Effects on Biological Hydrogen Production by a Mixed Culture

  • Jun, Yoon-Sun;Yu, Seung-Ho;Ryu, Keun-Garp;Lee, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • v.18 no.6
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    • pp.1130-1135
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    • 2008
  • The effect of pH on anaerobic hydrogen production was investigated under various pH conditions ranging from pH 3 to 10. When the modified Gompertz equation was applied to the statistical analysis of the experimental data, the hydrogen production potential and specific hydrogen production rate at pH 5 were 1,182 ml and 112.5 ml/g biomass-h, respectively. In this experiment, the maximum theoretical hydrogen conversion ratio was 22.56%. The Haldane equation model was used to find the optimum pH for hydrogen production and the maximum specific hydrogen production rate. The optimum pH predicted by this model is 5.5 and the maximum specific hydrogen production rate is 119.6 ml/g VSS-h. These data fit well with the experimented data($r^2=0.98$).

Enterococcus faecium 19-46-4에 의한 Cholic Acid의 생산

  • Chung, Eun-Young;Kim, Myung-Soo;Lee, Chul-Hoon;Kim, Byung-Hong
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.540-545
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    • 1996
  • A facultative anaerobe Enterococcus faecium 19-46-4 was used to study the production of an antimicrobial substance in anaerobic conditions. Major part of the antibiotic activity was found in the culture filtrate of the bacterium. The active compound was extracted by an equal volume of iso-butanol and concentrated in vacuo (at 50$\circ$C) before purification by C-18 liguid column chromatography and HPLC. A chromatographically pure compound was obtained by two passages of HPLC columns, The compound appeared as a pale-yellow powder. The yield was about 2.5 mg 1$^{-1}$ culture filtrate. The compound was named as KIST 194. KIST 194 were identified as cholic acid (3$\alpha$, 7$\alpha$, 12$\alpha$-trihydroxy-5$\beta$-cholan 24-oic acid) based on its physico-chemical properties determined by UV, IR, $^{1}H-NMR, $^{13}$C-NMR, El-MS and LC-MS.

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Culture Conditions and Growth Characteristics of Bifidobacterium longum

  • KIM, WANG-JUNE;SEONG-KWAN CHA
    • Journal of Microbiology and Biotechnology
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    • v.5 no.3
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    • pp.149-153
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    • 1995
  • A simple and low-cost medium was developed for the growth of Bifidobacterium longum KFRI 977. Of three bifidobacterial strains, B. longum KFRI 977 (ATCC 15707) showed the best growth in MRSC broth containing 0.3% oxgall, grew well in partially anaerobic condition, exhibited highest $\beta$-galactosidase activity, and was inhibitory against Clostridium perfringens KFRI 434. Of three developed media, the population of B. longum KFRI 977 was highest (1.9$\times$$10^9$/ml) in ISP based medium. The composition of ISP based medium is ISP (5%), glucose (1%), L-cysteine HCI (0.05%), Trypticase peptone (0.5%), yeast extract (0.5%), $MgSO_4$ (0.05%), Tween-80 (0.1%), and phosphate buffer (pH 7.0). Hydrolysis of ISP by Protease A was unnecessary, and the use of phosphate buffer (pH 7.0) prevented the formation of protein precipitate. Associative culture of B. longum KFRI 977 with Lactobacillus acidophilus KFRI 233 was proven to be deleterous to the growth of B. longum KFRI 977.

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Study on the Fermentation Conditions Influencing the Production of Vitamin $B_{12}$ by Propionibacterium shermanii (Propionibacterium shermanii에 의한 Vitamin $B_{12}$생성에 영향을 미치는 발효조건에 관한 연구)

  • 김지영;김공환구양모
    • KSBB Journal
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    • v.7 no.2
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    • pp.126-131
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    • 1992
  • The effects of fermentation conditions and medium compositions on the production of vitamin $B_{12}$ by Propionibacterium shermanii IFO 1239 were studied. Changes from an anaerobic to aerobic condition and a complex to synthetic medium after 48hr resulted in a 100% increase in vitamin $B_{12}$ production compared to an anaerobic culture alone. Glucose, fructose and lactose were found to be equally good as a carbon source for vitamin $B_{12}$ production. Addition of succinate and malate to the synthetic medium with glucose as a carbon source led to an increase in vitamin $B_{12}$ production by 33.6% and 17.2% respectively.

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Metabolic Flux Distribution in a Metabolically Engineered Escherichia coli Strain Producing Succinic Acid

  • Hong, Soon-Ho;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.10 no.4
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    • pp.496-501
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    • 2000
  • Escherichia cole NZN111, which is known as a pfl ldhA double mutant strin, was metabolically engineered to produce succinic acid by overexpressing malic enzyme into the E. coli controlled by a trc promoter. Fermentation studies were carried out in a LB medium by first growing cells aerobically to an $OD_{600}$ of 5. At this point, 0.01 mM IPTG was added to induce the overexpression of malic enzyme and the agitation speed was gradually lowered. When the culture $OD_{600}$ reached 11, a complete anaerobic condition was achieved by flushing with a $CO_3-H_2$ gas mixture. When NZN111(pTrcML) was cultured at $37^{\circ}C$, the final succinic acid concentration of 2.8 g/l could be obtained after 30 h of anaerobic cultivation. The fermentation results were analyzed by the calculation of metabolic fluxes. Metaolic flux analysis showed that about 85% of phosphoenolpyruvate (PEP) was converted to pyruvate, and further converted to malic acid by malic enzyme.

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