• Title/Summary/Keyword: Microbial Culture

Search Result 884, Processing Time 0.03 seconds

Application of Polymerase Chain Reaction (PCR) to the Diagnosis of Tuberculosis (Polymerase Chain Reaction (PCR)을 이용한 결핵의 진단에 관한 연구)

  • Kim, Ho-Joong;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
    • /
    • v.39 no.6
    • /
    • pp.517-525
    • /
    • 1992
  • Background: Since its development by Saiki et al, polymerase chain reaction (PCR) has been very useful in various fields of molecular biology. PCR can be used for the detection of a very small amount of microbial agent, and is especially useful in those patients who are difficult to diagnose microbiologically or serologically. Mycobacterium tuberculosis is a very slowly growing organism and AFB staining frequently shows false negative results, and therefore PCR would be a very rapid, easy, and sensitive diagnostic method for the diagnosis of Mycobacterium tuberculosis. Method: To compare PCR with conventional methods in diagnosing Mycobacterium tuberculosis in sputum, we used sputa of patients who visited or were admitted to Seoul National University Hospital. The amplification targets were 383 base pair DNA, a part of 2520 base pair DNA encoding 65 kD Mycobacterium tuberculosis specific protein (the primers are TB-1, -2), and 123 base pair DNA, a part of IS6110 fragment, which multiple copies are known to exsist PCR one genome (the primers are Sal I-1, -2). We also requested AFB staing and culture to the lab of Seoul National University Hospital with the same sample and compared the results. Results: 1) Using TB-1, -2 primers, PCR was positive in 73.1% (19/26) of culture positive sputa, in 12.5% (1/8) of culture negative. but clinically diagnosed tuberculous sputa, and was negative in all sputa of patients who were clinically diagnosed as non-tuberculous etiology. 2) Using Sal I-I, -2 primers, PCR was positive in 94.1% (32/34) of culture positive sputa, in 23.1% (6/26) of culture negative, but clinically diagnosed tuberculous sputa, and was negative in 87.5% (14/16) of sputa from patients who were clinically diagnosed as non-tuberculous etiology. Conclusion: PCR could be a very rapid, sensitive and specific method for the diagnosis of Mycobacterium tuberculosis in sputa, and further studies should be followed for the development of easier method.

  • PDF

Phylogenetic Diversity of Bacteria in an Earth-Cave in Guizhou Province, Southwest of China

  • Zhou, Jun-Pei;Gu, Ying-Qi;Zou, Chang-Song;Mo, Ming-He
    • Journal of Microbiology
    • /
    • v.45 no.2
    • /
    • pp.105-112
    • /
    • 2007
  • The objective of this study was to analyze the phylogenetic composition of bacterial community in the soil of an earth-cave (Niu Cave) using a culture-independent molecular approach. 16S rRNA genes were amplified directly from soil DNA with universally conserved and Bacteria-specific rRNA gene primers and cloned. The clone library was screened by restriction fragment length polymorphism (RFLP), and representative rRNA gene sequences were determined. A total of 115 bacterial sequence types were found in 190 analyzed clones. Phylogenetic sequence analyses revealed novel 16S rRNA gene sequence types and a high diversity of putative bacterial community. Members of these bacteria included Proteobacteria (42.6%), Acidobacteria (18.6%), Planctomycetes (9.0 %), Chloroflexi (Green nonsulfur bacteria, 7.5%), Bacteroidetes (2.1%), Gemmatimonadetes (2.7%), Nitrospirae (8.0%), Actinobacteria (High G+C Gram-positive bacteria, 6.4%) and candidate divisions (including the OP3, GN08, and SBR1093, 3.2%). Thirty-five clones were affiliated with bacteria that were related to nitrogen, sulfur, iron or manganese cycles. The comparison of the present data with the data obtained previously from caves based on 16S rRNA gene analysis revealed similarities in the bacterial community components, especially in the high abundance of Proteobacteria and Acidobacteria. Furthermore, this study provided the novel evidence for presence of Gemmatimonadetes, Nitrosomonadales, Oceanospirillales, and Rubrobacterales in a karstic hypogean environment.

Low Ruminal pH Reduces Dietary Fiber Digestion via Reduced Microbial Attachment

  • Sung, Ha Guyn;Kobayashi, Yasuo;Chang, Jongsoo;Ha, Ahnul;Hwang, Il Hwan;Ha, J.K.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.2
    • /
    • pp.200-207
    • /
    • 2007
  • In vitro rumen incubation studies were conducted to determine effects of initial pH on bacterial attachment and fiber digestion. Ruminal fluid pH was adjusted to 5.7, 6.2 and 6.7, and three major fibrolytic bacteria attached to rice straw in the mixed culture were quantified with real-time PCR. The numbers of attached and unattached Fibrobacter succinogenes, Ruminococcus flavefaciens and Ruminocococcus albus were lower (p<0.05) at initial pH of 5.7 without significant difference between those at higher initial pH. Lowering incubation media pH to 5.7 also increased bacterial numbers detached from substrate regardless of bacterial species. Dry matter digestibility, gas accumulation and total VFA production were pH-dependent. Unlike bacterial attachment, maintaining an initial pH of 6.7 increased digestion over initial pH of 6.2. After 48 h in vitro rumen fermentation, average increases in DM digestion, gas accumulation, and total VFA production at initial pH of 6.2 and 6.7 were 2.8 and 4.4, 2.0 and 3.0, and 1.2 and 1.6 times those at initial pH of 5.7, respectively. The lag time to reach above 2% DM digestibility at low initial pH was taken more times (8 h) than at high and middle initial pH (4 h). Current data clearly indicate that ruminal pH is one of the important determinants of fiber digestion, which is modulated via the effect on bacterial attachment to fiber substrates.

Rapid and exact molecular identification of the PSP (paralytic shellfish poisoning) producing dinoflagellate genus Alexandrium

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
    • /
    • 2003.10a
    • /
    • pp.132-133
    • /
    • 2003
  • The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

  • PDF

Ruminal microbial responses in fermentation characteristics and dry matter degradability to TDN level of total mixed ration

  • Lee, Seung-Uk;Jo, Jin-Ho;Park, Sung-Kwon;Choi, Chang-Weon;Jeong, Jun;Chung, Ki-Young;Chang, Sun-Sik;Li, Xiang Zi;Choi, Seong-Ho
    • Korean Journal of Agricultural Science
    • /
    • v.43 no.1
    • /
    • pp.80-86
    • /
    • 2016
  • An in vitro trial was conducted to examine the effects of total mixed rations (TMR) on fermentation characteristics and effective degradability (ED) by rumen microbes. Three TMR diets were growing period TMR (GR-TMR, 67% TDN), early fattening period TMR (EF-TMR, 75.4% TDN) and late fattening TMR (LF-TMR, 80% TDN). Three TMR diets (3 g of TMRs in each incubation bottles) was added to the mixed culture solution of stained rumen fluid with artificial saliva (1:1, v/v) and incubated anaerobically for 48 hours at $39^{\circ}C$. The pH in all incubation solutions tended to decrease up to 48h, but the opposite results were found in concentration of total gas production, ammonia-N and total VFA in all incubations.The total gas production (p<0.05) in LF-TMR was highest compared with those of other diets. Also, concentration of total VFA was tended to increase in LF-TMR compared with other TMR diets in all incubations. The EDDM in both EF-TMR and LF-TMR was tended to high compared with GR-TMR (p=0.100). In this in vitro trials, concentration of propionate in all incubation solution was not affected by increased concentration of TDN. The results of the present in vitro study indicate that TMR may provide more favorable condition for nutrient digestion both in the rumen.

Isolation and characterization of microorganisms biological damage of Dongchundang (동춘당 생물학적 가해 미생물의 분리 및 특성)

  • Lee, Jeung-Min;Kim, Young-Hee;Hong, Jin-Young;Jo, Chang-Wook;Kim, Soo Ji;Seo, Min Seok
    • 보존과학연구
    • /
    • s.35
    • /
    • pp.111-119
    • /
    • 2014
  • Microorganisms were isolated from Dongchundang(wooden cultural heritage) with PDA medium culture. Nineteen species shows the cellulolytic activity. Methylobacterium sp. was the most active in cellulose degradation. The growth curve and pH were measured during incubation of the microorganism for 72 hours. The pH was increased with the increasing of microbial growth. The degree of cellulose degradation was determined with the amount of reducing sugar by use of dinitrosalicylic acid (DNS) method. The amount of reducing sugar was decreased after 45 hours. As a results, It should suggested that wood component were deteriorated by Methylobacterium sp..

  • PDF

Antagonistic Bacillus species as a biological control of ginseng root rot caused by Fusarium cf. incarnatum

  • Song, Minjae;Yun, Hye Young;Kim, Young Ho
    • Journal of Ginseng Research
    • /
    • v.38 no.2
    • /
    • pp.136-145
    • /
    • 2014
  • Background: This study aimed to develop a biocontrol system for ginseng root rot caused by Fusarium cf. incarnatum. Methods: In total, 392 bacteria isolated from ginseng roots and various soils were screened for their antifungal activity against the fungal pathogen, and a bacterial isolate (B2-5) was selected as a promising candidate for the biocontrol because of the strong antagonistic activity of the bacterial cell suspension and culture filtrate against pathogen. Results: The bacterial isolate B2-5 displayed an enhanced inhibitory activity against the pathogen mycelial growth with a temperature increase to $25^{\circ}C$, produced no pectinase (related to root rotting) an no critical rot symptoms at low [$10^6$ colony-forming units (CFU)/mL] and high ($10^8CFU/mL$) inoculum concentrations. In pot experiments, pretreatment with the bacterial isolate in the presumed optimal time for disease control reduced disease severity significantly with a higher control efficacy at an inoculum concentration of $10^6CFU/mL$ than at $10^8CFU/mL$. The establishment and colonization ability of the bacterial isolates on the ginseng rhizosphere appeared to be higher when both the bacterial isolate and the pathogen were coinoculated than when the bacterial isolate was inoculated alone, suggesting its target-oriented biocontrol activity against the pathogen. Scanning electron microscopy showed that the pathogen hyphae were twisted and shriveled by the bacterial treatment, which may be a symptom of direct damage by antifungal substances. Conclusion: All of these results suggest that the bacterial isolate has good potential as a microbial agent for the biocontrol of the ginseng root rot caused by F. cf. incarnatum.

Protection of Tobacco Plants from Bacterial Wilt with an Avirulent Isolate of Pseudomonas solanacearum (비병원성 Pseudomonas solanacearum을 이용한 담배 세균성마름병의 방제)

  • Yi Y. K.;Kim J. H.;Park W. M.
    • Korean Journal Plant Pathology
    • /
    • v.1 no.1
    • /
    • pp.17-21
    • /
    • 1985
  • Microbial antagonism between virulent and avirulent isolates of Pseudomonas solanacearum was studied in relation to the control of bacterial wilt of tobacco. In nutrient broth media or in soil, the avirulent isolate of P. solanacearum grew faster than did the virulent one. Inhibitory effect of avirulent isolate against growth of virulent one was negligible in mixed culture of the two isolates. The disease severity of bacterial wilt was significantly reduced when the roots of cultivar BY 4 susceptible to bacterial wilt was dipped in suspension of an avirulent isolate for 6 hours prior to transplanting to the soil infested with virulent bacteria. When the seedlings of tobacco were poured with the suspension of an avirulent isolate onto the soil in pre-planting pots 24 hours before ransplanting, there was a significant reduction in disease severity in the field. However, the reduction was noticed until early July, but after middle of July, no difference between the avirulent isolate-treated and non-treated plants was found in severity of the bacterial wilt.

  • PDF

Incorporation of silver nanoparticles on the surface of orthodontic microimplants to achieve antimicrobial properties

  • Venugopal, Adith;Muthuchamy, Nallal;Tejani, Harsh;Anantha-Iyengar-Gopalan, Anantha-Iyengar-Gopalan;Lee, Kwang-Pill;Lee, Heon-Jin;Kyung, Hee Moon
    • The korean journal of orthodontics
    • /
    • v.47 no.1
    • /
    • pp.3-10
    • /
    • 2017
  • Objective: Microbial aggregation around dental implants can lead to loss/loosening of the implants. This study was aimed at surface treating titanium microimplants with silver nanoparticles (AgNPs) to achieve antibacterial properties. Methods: AgNP-modified titanium microimplants (Ti-nAg) were prepared using two methods. The first method involved coating the microimplants with regular AgNPs (Ti-AgNP) and the second involved coating them with a AgNP-coated biopolymer (Ti-BP-AgNP). The topologies, microstructures, and chemical compositions of the surfaces of the Ti-nAg were characterized by scanning electron microscopy (SEM) equipped with energy-dispersive spectrometer (EDS) and X-ray photoelectron spectroscopy (XPS). Disk diffusion tests using Streptococcus mutans, Streptococcus sanguinis, and Aggregatibacter actinomycetemcomitans were performed to test the antibacterial activity of the Ti-nAg microimplants. Results: SEM revealed that only a meager amount of AgNPs was sparsely deposited on the Ti-AgNP surface with the first method, while a layer of AgNP-coated biopolymer extended along the Ti-BP-AgNP surface in the second method. The diameters of the coated nanoparticles were in the range of 10 to 30 nm. EDS revealed 1.05 atomic % of Ag on the surface of the Ti-AgNP and an astounding 21.2 atomic % on the surface of the Ti-BP-AgNP. XPS confirmed the metallic state of silver on the Ti-BP-AgNP surface. After 24 hours of incubation, clear zones of inhibition were seen around the Ti-BP-AgNP microimplants in all three test bacterial culture plates, whereas no antibacterial effect was observed with the Ti-AgNP microimplants. Conclusions: Titanium microimplants modified with Ti-BP-AgNP exhibit excellent antibacterial properties, making them a promising implantable biomaterial.

Fermentative Hydrogen Production from the Pretreated Food-Processing Waste and Sewage Sludge using Chemical/Ultra-Sonication (두부제조폐기물과 하수슬러지의 화학/초음파 전처리에 의한 가용화 및 혐기발효 수소생산)

  • Kim, Mi-Sun;Lee, Dong-Yeol;Kim, Dong-Hun;Kim, Ok-Sun;Lim, So-Yung
    • Transactions of the Korean hydrogen and new energy society
    • /
    • v.21 no.6
    • /
    • pp.580-586
    • /
    • 2010
  • Acid and alkali pretreatments were applied to tofu processing waste (TPW) to increase the solubility of ingredients in TPW. Pretreatment at 1.0% of HCl and 2.5% of NaOH condition resulted in the increase of SCOD concentration from 3.2 g COD/L to 27 g COD/L and 33 g COD/L, respectively. The acid and alkali-pretreated TPW was studied for its fermentative $H_2$ production capacity in batch mode using a thermophillic mixed culture. Alkali pretreatment on presence of 2.5% NaOH exhibited more soluble portion released compared to acid pretreatment using HCl, however the $H_2$ production from acid pretreated TPW was better than alkali-pretreated TPW probably due to the sodium inhibition on microbial activity. In addition, sewage sludge was externally added to the acid-pretreated (1.0% HCl) TPW by 20% (on volume basis). Average H2 production rate was increased from 31 to 78 ml/L-broth/hr, and it was attributed to the high buffer capacity and abundant nutrients especially divalent cation in sewage sludge.