• Title/Summary/Keyword: anaerobic incubator

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Development of a Bottle-Free Multipurpose Incubator for Generating Various Bacterial Culture Conditions

  • Yang, Nam-Woong;Lim, Yong
    • Journal of Microbiology
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    • v.43 no.1
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    • pp.28-33
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    • 2005
  • The purpose of this study was to develop a multipurpose incubator, without the gas cylinders (bottles) which are required for $H_2$ and $CO_2$ supplementation. In our bottle-free multipurpose incubator, the $H_2$ and $CO_2$ were generated by chemical reactions induced within the chamber. The reaction between sodium borohydride and acetic acid at a molar ratio of 1:1 was used to generate $H_2$, according to the following formula: $4NaBH_4+2CH_3COOH+7H_2O{\rightarrow} 2CH_3COONa+Na_2B_4O_7+16H_2$, whereas the other reaction, citric acid and sodium bicarbonate at a 1:1 molar ratio, was used to generate $CO_2$, according to the following formula: $C_6H_8O_7+3NaHCO_3{\rightarrow}Na_3(C_6H_5O_7)+3H_2O+3CO_2$. Five species of obligate anaerobic bacteria, one strain of capnophilic bacterium, and one strain of microaerophilic bacterium were successfully cultured in the presence of their respective suitable conditions, all of which were successfully generated by our bottle-free multipurpose incubator. We conclude that, due to its greater safety, versatility, and significantly lower operating costs, this bottle-free multipurpose incubator can be used for the production of fastidious bacterial cultures, and constitutes a favorable step above existing anaerobic incubators.

EFFECT OF POLYPHOSPHATE IN ROOT CANAL SEALERS ON THE GROWTH OF ORAL BACTERIA (Polyphosphate가 함유된 근관충전재가 구강세균의 성장에 미치는 영향)

  • 박석범;최기운;최호영
    • Restorative Dentistry and Endodontics
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    • v.26 no.2
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    • pp.141-152
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    • 2001
  • Eliminating the infecting bacteria of the root canal system and preventing reinfection must be the main objectives of all endodontic works. None of commercially available root canal sealers have the properties of desirable tissue compatibility and strong antibacterial activity. The purpose of this study is to develope an ideal root canal sealer using commercially available polyphosphate (polyP), Calgon, which is known to be antibacterial and safe. For the study. resin type AH26, zinc oxide eugenol type Tubli Seal. Ca(OH)$_2$ type Apexit as base sealers for polyP (0~3%) and para formaldehyde containing N2 as a control base were selected. Specimens (3$\times$4mm) of the sealers were prepared in a 37$^{\circ}C$ incubator for 3 and 10 days and their antibacterial activity against streptococci and black pigmented anaerobic rods was observed using an agar diffusion method. The result were as follows: 1. Among 3 day old root canal sealers. N2 as a positive control showed the strongest antibacterial effect. followed by AH26. Tubli Seal and. Apexit which barely showed antibacterial activity against the test bacteria. In contrast. 10 day old AH26 showed a greater antibacterial activity than 10 day old N2. 2. All sealer specimens showed a greater antibacterial activity against black pigmented anaerobic rods than streptococci. Three day old ones appeared to be more antibacterial than 10 day old ones except for Apexit. 3. As compared to N2, 3 day old AH26 demonstrated a similar antibacterial activity against black pig mented anaerobic rods but to a lesser extent to streptococci. Ten day old AH26 showed a greater antibacterial activity against black pigmented anaerobic rods than 10 day old N2. 4. As compared to AH26. Tubli Seal generally revealed a lower antibacterial activity but it showed a greater antibacterial activity aginst S. gordonii Challis. 5. Enhancement of antibacterial activity by polyP was more clearly observed when it was added to Ca(OH)$^{\circ}C$ based root canal sealers. Tubli Seal and N2. 6. The addition of polyP enhanced the antibacterial activity of 3 day old AH26 against S. gordonii G9B (16%) and Challis (29%), and P. gingivalis 2561 (24%) only. Moreover, polyP failed to increase antibacterial activity of 10 day old AH26 against the test strains but P. gingivalis A7A1 28(13%). 7. The addition of polyP increased the antibacterial effect of 3 day old Tubli Seal on several test bacteria including s. mutans GS 5 (50%). s. gordonii G9B (47%) and Challis (122%). and all the test strains of P. gingivalis (13~35%) except for 9 14K 1. The addition of polyP to 10 day old Tubli Seal increased antibacterial activity of the root canal sealer against most test strains. 8. 3 day old Apexit failed to show antibacterial activity. if any very little against S. mutans GS 5 and Pr. intermedia ATCC 49046. However. polyP increased its antibacterial activity by 50 and 69%, respectively. Increase of antibacterial activity of 10 day old Apexit by polyP was more clearly observed than that of 3 day old one.

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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.

The Interactive Effect of These Bacterial Substrates on the Growth of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis (Streptococcus gordonii, Fusobacterium nucleatum 및 Porphyromonas gingivalis의 상호작용이 성장에 미치는 영향)

  • Kim, A-Reum;Jeong, Moon-Jin;Ahn, Yong-Soon;Kim, Mi-Na;Kim, Sung-Im;Lim, Do-Seon
    • Journal of dental hygiene science
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    • v.15 no.2
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    • pp.209-219
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    • 2015
  • In order to explore an effect of interaction of Streptococcus gordonii, Fusobacterium nucleatum and Porphyromonas gingivalis that are bacteria relevant to periodontal disease on its growth, the bacteria were incubated in trypticase soy hemin menadione broth at $37^{\circ}C$ $CO_2$ incubator for 7 days through anaerobic jar by single and co-culture with heat treated dead bacteria under anaerobic gas pack. In order to confirm growth level, absorbance was measured and for confirming colony structure and form, it was observed with scanning electron microscope. In order to confirm an effect on pathogenicity of P. gingivalis, real time reverse transcriptase polymerase chain reaction was implemented for expression analysis for rgpA gene that produces HRgpA which is gingipain. As a result, the following conclusion was obtained. Colony formation of S. gordonii and P. gingivalis was increased by other dead bacteria and in case of F. nucleatum, its colony formation was showed an aspect of being increased by dead bacterium of P. gingivalis but decreased by dead bacterium of S. gordonii. Therefore, it is considered that the strains being used for this study would affect interactively through bacterial cell itself as well as their interaction factor at the time of colony formation.

Outbreaks of Strangles due to Capnophilic Streptococcus equi subsp equi in South Korea (호이산화탄소성 Streptococcus equi subspecies equi에 의한 선역의 발생)

  • Moon, Ja-Ho;Kim, Jeong-Hee;Takai, Shinji;Son, Won-Geun
    • Journal of Veterinary Clinics
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    • v.32 no.1
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    • pp.41-44
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    • 2015
  • We reported an outbreak of clinical strangles in thoroughbred horses due to capnophilic Streptococcus equi subsp equi in South Korea. On three different farms, we isolated 17 S equi subsp equi isolates from 29 horses with or without abscesses in their lymph nodes. Of the 17 isolates, two isolates from clinical cases grew well in aerobic conditions, whereas 7/7 isolates from clinical cases and 8/22 isolates from the nasal discharges of horses did not. The latter 15 isolates were capnophilic, oxygen-sensitive, and $CO_2$-requiring S equi subsp equi, which could not grow in aerobic conditions, but which grew well in a $CO_2$ incubator with 5% $CO_2$, in anaerobic conditions using a GasPak, and with reduced oxygen tension in a candle jar. This study is the first report of a strangles outbreak caused by capnophilic S equi subsp equi in South Korea.

MOLECULAR IDENTIFICATION OF BACTERIA FROM OSTEOMYELITIS OF THE JAWS (분자생물학적 기법을 이용한 악골 골수염 병소의 세균 동정)

  • Kim, Mi-Sung;Kim, Su-Gwan;Chung, Hae-Man;Kim, Sang-Gon;Kook, Joong-Ki;Kim, Mi-Kwang;Kim, Hwa-Sook;Yoo, So-Young
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.29 no.1
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    • pp.48-55
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    • 2003
  • The purpose of this study was to isolate and identify the bacteria in osteomyelitis lesion of 3 patients. Two lesions were due to the post-infection after extraction. The other was resulted from mal-fixation of both sides of mandibular angles. Pus samples were collected by needle aspiration from the lesion and examined by culture method. Bacterial culture was performed in three culture systems (anaerobic, $CO_2$, and aerobic incubator). Identification of the bacteria was performed by 16S rRNA gene cloning and nucleotide sequencing method. Our results showed that Streptococci species was predominantly isolated in both lesions of extraction socket. Only one species (Proteus vulagris) was detected in lesion of mandibular angle. This study was not sufficient to identify the causative bacteria in those osteomyelitis. However, our data may be offered the clue to solve the problem.

The Changes of Intestinal Normal Flora in Neonates for Seven Days Postnatally (정상 신생아의 대변에서 생후 1주일 동안 장내세균총의 변화)

  • Sung, Nam-ju;Lee, Seung Gue;Kim, Me Jin;Kim, Young Ho;Yang, Seung;Hwang, Il Tae;Jung, Ji A;Lee, Hae Ran;Kim, Jae-Seok
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.9 no.2
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    • pp.162-168
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    • 2006
  • Purpose: Microbial colonization of the intestine begins just after birth and development of the normal flora is a gradual process. The first bacteria colonizing the intestine in newborns are Staphylococcus, Enterobacteriaceae and Streptococcus. For several days after birth, the number of Bifidobacterium spp. increase. The aim of this study was to investigate the changes of microflora for seven days postnatally in neonatal stool. Methods: Fifteen neonates (breast : formula : mixed feeding 1 : 8 : 6, vaginal delivery : cesarean section 3 : 12) who were born at the Kangdong Sacred Heart Hospital, Hallym University were enrolled. First meconium and stools of postnatal 1-, 3-, and 7-day were innoculated. Blood agar plates for total aerobes, trypton bile X-glucuronide agar for E. coli, phenylethyl alcohol agar for gram positive anaerobes, MRS agar for Lactobacillus spp., bifidobacterium selective agar for Bifidobacterium spp. and cefoxitin-cycloserine-fructose agar for Clostridium difficile were used in the general incubator ($CO_2$ free incubator), $CO_2$ incubator or the anaerobic chamber for 48 or 72 hours at $37^{\circ}C$ and then colony forming units were counted. Results: No microflora was identified in the first meconium. Total aerobes, E. coli, and gram positive anaerobes were significantly increased with advancing postnatal days. In only one baby, Lactobacillus acidophilus was detected $2{\times}10^5CFU/g$ in the seven-day stool. Bifidobacterium spp. was detected in two babies. Clostridium difficile was not detected during the seven days. There were no significant differences in the bowel flora depending on the delivery pattern and feeding method. Conclusion: This study shows many changes in the intestinal normal flora in neonatal stool during seven days postnatally. If these findings are confirmed with larger studies, the data may be preliminary findings to support use of probiotics in neonates.

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FACTORS AFFECTING THE FORMATION OF SOLUBLE GLUCAN BY LACTOCOCCUS LACTIS 1370 (유산구균 1370의 수용성 글루캔 형성에 영향을 미치는 인자)

  • Oh, In-Gyun;Yang, Kyu-Ho;Chung, Jin;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.2
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    • pp.185-191
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    • 2000
  • The water-soluble glucan produced by Lactococcus lactis 1370 affects the formation of dental plaque by Streptococcus mutans. In this study the factors affecting the formation of water-soluble glucan were assessed as the optical density of culture supernatant of Lactococcus lactis 1370 in the spectrophotometer. 1. The optical density of culture supernatant was high when Lactococcus lactis 1370 was cultured in M17 broth with 5% sucrose, while being low in culture supernatant of Streptococcus mutans. 2. The optical density of culture supernatant was higher when Lactococcus lactis 1370 was cultured in M17 broth with 10% sucrose than when being cultured without sucrose (p<0.05), and was higher at pH 7 than pH 5 (p<0.05). 3. The optical density of culture supernatant was the highest at $37^{\circ}C$ among $32^{\circ}C,\;37^{\circ}C\;and\;42^{\circ}C$, and was higher in the anaerobic incubator than in the aerobic incubator (p<0.05). 4. The optical density of culture supernatant was the highest in the media containing 1.0mM $CaCl_2$ (p<0.05), 2.5mM KCl (p<0.05), and 1.6mM $MgCl_2$. 5. When Streptococcus mutans was cultured in the media containing a quarter culture supernatant of Lactococcus lactis 1370 grown in M17 broth, the mean weight of produced artificial plaque was 103.0mg on the wire, whereas being significantly reduced to 5.6mg in the media containing a quarter culture supernatant of Lactococcus lactis 1370 grown in M17 broth containing 5% sucrose (p<0.05). These results indicate that the water-soluble glucan is more formed by Lactococcus lactis 1370 in the media containing sucrose or under the adequate conditions for the growth of bacteria, and inhibits the formation of artificial plaque by Streptococcus mutans.

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The protective effect of hypoxic therapy on paraquat-induced toxicity rat model (백서를 이용한 고독성 제초제 파라쿼트 중독 치료를 위한 저농도산소요법의 효과 연구)

  • Kim, Hoon;Min, Jin-Hong;Han, Kyu-Hong;Kang, Joon-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.4
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    • pp.2189-2198
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    • 2014
  • Paraquat (PQ) is a very effective and widely used herbicide that was commercially introduced in 1962. In this study, instead of using antioxidants like in the past, to inhibit the formation of PQ-induced ROS, we attempted to reduce the oxygen concentration by using non-lethal hypoxia therapy. Therefore, we studied the toxicity of PQ in vivo, analyzed the major effects of ROS on the targeted lung tissue and compared the results with the gross histological changes after the cell protective effect of non-lethal hypoxia therapy. In vivo studies demonstrated that low-concentration oxygen therapy (i.e., 10-12% oxygen) in rats administered with PQ was associated with a higher survival rate than in rats that received only PQ. In vivo non-lethal hypoxia treatment showed better survival and less lung tissue damage. Using a hypoxic/anaerobic incubator with integrated multifaceted molecular analysis, including MDA assay, glutathione assay, and SOD assay, we established an optimal, significantly reduced in vivo non-lethal hypoxia treatment by exploiting the PQ-induced cytotoxicity responses.

Preparation of fermentation broth of Sparassis latifolia containing soluble β-glucan using four Lactobacillus species (수용성 β-glucan을 함유한 꽃송이버섯 발효액의 제조)

  • Jo, Han-Gyo;Choi, Moon-Hee;Shin, Hyun-Jae
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.50-55
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    • 2015
  • Glucan has been shown to have a significant role in the activation of the immune system, including increased activity of macrophages and so on. Sparassis latifolia (formerly S. crispa) is an edible mushroom abundant in dietary fiber and widely known to contain high levels of ${\beta}$-glucan. In the present study, fermentation broths containing soluble ${\beta}$-glucan were prepared by fermentation with mushrooms with four Lactobacillus species (L. plantarum subsp. Plantarum, L. acidophilus, L. helveticus, and L. delbrueckii subsp. Bulgaricus). After culturing four Lactobacillus spp. in MRS broth, each Lactobacillus was inoculated into MRS broth containing S. latifolia powder 5% (w/v) at $37^{\circ}C$ in an anaerobic incubator for five days. It showed that the ${\beta}$-glucan contents were different in each fermentation sample. The suitable conditions for the preparation of mushroom fermentation broths were investigated and discussed.