• Title/Summary/Keyword: enrichment culture

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Isolation and Growth Pattern of Bacillus cereus from Ready-to-Eat Foods. (즉석조리식품에서의 Bacillus cereus오염실태조사 및 생육 pattern 분석)

  • 김순한;김미경;강민철;손영욱;이창희;김인복;이영자;최수영
    • Journal of Life Science
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    • v.14 no.4
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    • pp.664-669
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    • 2004
  • The contamination of Bacillus cereus was investigated in 240 RTE (ready-to-eat) food samples including 118 seafoods, 82 Korean packaged meals and 40 other RTE foods. Many B. cereus presumptive strains were isolated from the enrichment culture in Tryptic Soy Broth (TSB) added polymyxin, followed by selective culture in Mannitol Egg Yolk Polymyxin (MYP) agar and Gram staining. A total of 36 strains (16 in seafoods, 17 in Korean pack-aged meals and 3 in other RTE foods) were identified as B. cereus by the analysis of 61 biochemical tests of the API 50CHB/20E system test and supplementary tests of $\beta$-hemolysis, rhizoid growth, motility and oxidase activity. The 28 strains out of 36 B. cereus isolates produced diarrhoeal enter-otoxin in Brain Heart Infusion (BHI) broth. All isolates were resistant to ampicillin and penicillin antibiotics, and most of them were susceptible to gentamicin, vancomycin, bacitracin, chloram-phenicol, kanamycin and streptomycin. The growth of B. cereus was affected by environmental temperature and incubation time. Culture with temperature under 1$0^{\circ}C$ effectively restricted the growth of B. cereus.

Clean-up of the Crude Oil Contaminated Marine Sediments Through Biocarrier-Mediated Bioaugmentation (생물담체 활용 생물접종에 의한 원유로 오염된 해양토양의 정화)

  • Ekpeghere, Kelvin I.;Bae, Hwan-Jin;Kwon, Sung-Hyun;Kim, Byung-Hyuk;Park, Duck-Ja;Kim, Hee-Shik;Koh, Sung-Cheol
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.354-361
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    • 2009
  • This study was carried out to develop an effective biocarrier-mediated bioaugmentation technology which will be useful for remediation of the crude oil-contaminated marine sediments. Enrichment of several microbial communities was made from several oil-polluted seashore sites and the two distinctively functional consortia have been successfully selected. These two consortia were grown together and used to manufacture the microbial agents for bioaugmentation of marine sediments polluted with crude oil. The most dominant species in the mixed culture was identified as Alcanivorax borkumensis based on pure culture and DGGE analysis. Bioaugmentation of oil-polluted marine sediments with the microbial agent MA-2 formulated using the mixed culture and biocarriers (activated carbon and minerals) was more effective, especially in combination with an oxygen producing (releasing) compound (ORC). Ninty percent of TPH was removed in the presence of ORC in 35 days while 74% in the absence of ORC. This indicated that the indigenous consortial degraders could be immobilized on the active carbon as a biocarrier to manufacture microbial agents and then effectively bioaugmented for remediation of the oil-polluted sediments.

The Study on the Paradigm Change of Policy Making in the field of Korean Content Industry (문화콘텐츠 정책평가와 개선방안에 관한 연구)

  • Shim, Sang-Min
    • Review of Culture and Economy
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    • v.17 no.2
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    • pp.103-135
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    • 2014
  • This research dwells on the new paradigm of policy making of some innovative nations in Korean content industry. Especially this paper did evaluation of governmental policy in recent 5 years (2008~2012) in the field of Korean content industry. According to this research, the program of policy in Korean content industry had been obsessed to passive enrichment focusing on some part of content industry. The whole process of policy, scheme of policy and working flow of policy were very constrained in order to encourage more measurable area like CT(culture technology). Thus, we need new strong policy in this new government launched in 2013. The apparent keyword should be 'management'. New activity of policy need to focus on encourage Korean content industry in the real site of field, not in bureaucratic office in remote site. This change reflecting real filed management system would be productive innovation for policy making and activity in Korea.

Edible Culture Media from Cereals and Soybeans for Pre-cultivation of Lactic Acid Bacteria (곡류 및 두류를 이용한 젖산균 전배양용 식용 배지의 제조)

  • Park, So-Lim;Park, Sunhyun;Jang, Jieun;Yang, Hye-Jung;Moon, Sung-Won;Lee, Myung-Ki
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.6
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    • pp.991-995
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    • 2013
  • This study was conducted to develop an edible culture media with various types of cereals and soybeans for the pre-cultivation of lactic acid bacteria (LAB). To manufacture the edible culture media, LAB enrichment media were prepared using cereals such as brown rice (including germinated brown rice, glutinous brown rice, and germinated glutinous brown rice), yellow soybeans (including yellow soybeans, hulled yellow soybeans, germinated yellow soybeans, hulled and germinated yellow soybeans), and black soybeans (black soybeans, hulled black soybeans, germinated black soybeans, hulled and germinated black soybeans). Seven species of LAB were used in the experiment: Lactobacillus (Lb.) farciminis, Lb. homohiochii, Lb. pentosus, Lb. plantarum, Leuconostoc (Leu.) paramesenteroides, Leu. citreum, and Leu. lactis. For edible culture media from cereals, the average viable cell count of the seven starter cultures was 7.6~8.0 log CFU/mL, while that of the MRS culture medium, a synthetic medium, was 9.2 log CFU/mL; thus proliferation was lower by about 1~2 log CFU/mL in starter cultures from cereals compared to the synthetic medium. In the case of the edible culture media from soybeans, most bacteria showed higher proliferation in the hulled and germinated soybean media. In particular, Lb. plantarum showed the highest cell count at 10.08 log CFU/mL. In the case of edible culture media from black soybeans, the proliferation rate was higher in the hulled and germinated black soybean medium. Lb. homohiochii showed the highest proliferation in the hulled and germinated black soybean medium at 9.90 log CFU/mL. All results show that edible culture media using cereals and soybeans are generally good for LAB. Especially, hulled and germinated black soybeans are optimal for the pre-cultivation of LAB medium.

Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Biodegradation of Diazinon by Serratia marcescens DI101 and its Use in Bioremediation of Contaminated Environment

  • Abo-Amer, Aly E.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.1
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    • pp.71-80
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    • 2011
  • Four diazinon-degrading bacteria were isolated from agricultural soil by using an enrichment technique. The biochemical analysis and molecular method including RFLP indicated that these isolates were identical, and one strain designated DI101 was selected for further study. Phylogenetic analysis based on 16S rDNA sequencing indicated that the strain DI101 clearly belongs to the Serratia marcescens group. The ability of the strain to utilize diazinon as a source of carbon and phosphorus was investigated under different culture conditions. The DI101 strain was able to completely degrade 50 mg/l diazinon in MSM within 11 days with a degradation rate of 0.226 $day^{-1}$. The inoculation of sterilized soil treated with 100 mg/kg of diazinon with $10^6$ CFU/g DI101 resulted in a faster degradation rate than was recorded in non-sterilized soil. The diazinon degradation rate by DI101 was efficient at temperatures from 25 to $30^{\circ}C$ and at pHs from 7.0 to 8.0. The degradation rate of diazinon was not affected by the absence of a phosphorus supplement, and addition of other carbon sources (glucose or succinate) resulted in the slowing down of the degradation rate. The maximum degradation rate ($V_{max}$) of diazinon was 0.292 $day^{-1}$ and its saturation constant ($K_s$) was 11 mg/l, as determined by a Michaelis-Menten curve. The strain was able to degrade diethylthiophosphate-containing organophosphates such as chlorpyrifos, coumaphos, parathion, and isazofos when provided as a source of carbon and phosphorus, but not ethoprophos, cadusafos, and fenamiphos. These results propose useful information for the potential application of the DI101 strain in bioremediation of pesticide-contaminated environments.

High Efficiency Binding Aptamers for a Wide Range of Bacterial Sepsis Agents

  • Graziani, Ana Claudia;Stets, Maria Isabel;Lopes, Ana Luisa Kalb;Schluga, Pedro Henrique Caires;Marton, Soledad;Ferreira, Ieda Mendes;de Andrade, Antero Silva Ribeiro;Krieger, Marco Aurelio;Cardoso, Josiane
    • Journal of Microbiology and Biotechnology
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    • v.27 no.4
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    • pp.838-843
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    • 2017
  • Sepsis is a major health problem worldwide, with an extremely high rate of morbidity and mortality, partly due to delayed diagnosis during early disease. Currently, sepsis diagnosis requires bacterial culturing of blood samples over several days, whereas PCR-based molecular diagnosis methods are faster but lack sensitivity. The use of biosensors containing nucleic acid aptamers that bind targets with high affinity and specificity could accelerate sepsis diagnosis. Previously, we used the systematic evolution of ligands by exponential enrichment technique to develop the aptamers Antibac1 and Antibac2, targeting the ubiquitous bacterial peptidoglycan. Here, we show that these aptamers bind to four gram-positive and seven gram-negative bacterial sepsis agents with high binding efficiency. Thus, these aptamers could be used in combination as biological recognition elements in the development of biosensors that are an alternative to rapid bacteria detection, since they could provide culture and amplification-free tests for rapid clinical sepsis diagnosis.

Biodegradation Kinetics of Nonylphenol Ethoxylates by Pseudomonas sp. (Pseudomonas sp.에 의한 Nonylphenol Ethoxylates의 Kinetics)

  • 김수정;이종근;이상준
    • Journal of Environmental Science International
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    • v.2 no.4
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    • pp.271-278
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    • 1993
  • Optimal biodegradation kinetics models to the initial nonylphenol ethoxylates-30 concentration were investigated and had been fitted by the linear regression. Microorganisms capable of degrading nonylphenol ethoxylates-30 were isolated from sewage near Ulsan plant area by enrichment culture technique. Among them, the strain designated as EL-10K had the highest biodegradability and was identified as Pseudomonas from results of taxonomical studies. The optimal conditions for the biodegradation were 1.0 g/ι of nonylphenol ethoxylates-30 and 0.02 g/ι of ammonium nitrate at pH 7.0 and 3$0^{\circ}C$. The highest degradation rate of nonylphenol ethoxylates-30 was about 89% for 30 hours incubation on the optimal condition. Biodegradation data were fit by linear regression to equations for 3 kinetic models. The kinetics of biodegradation of nonylphenol ethoxylates was best described by first order model for 0.1 $\mu\textrm{g}$/ι nonylphenol ethoxylates-30 ; by Monod no growth model and Monod with growth model for 0.5 $\mu\textrm{g}$/mι and 1.0, 5.0 $\mu\textrm{g}$/mι, respectively.

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Isolation and Degradation Characteristics of 2,4,4l-Trichloro-2l-Hydroxydiphenyl Ether Degrading Bacterium (2,4,4l-Trichloro-2l-Hydroxydiphenyl Ether 분해균의 분리 및 분해특성)

  • Han, Nan-Sook;Son, Hong-Joo;Lee, Geon;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.6 no.2
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    • pp.173-182
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    • 1997
  • The bacterial strains, which utilizes 2,4,4'-trichloro-2'-hydroxydiphenyl ether(TCHDPE) as a sole carbon source, were isolated by selective enrichment culture from soil samples of industrial waste deposits. The bacterium that showed the highestt biodegradation activity was designated as EL-O47R The isolated strain EL-O47R was Identified as the genus Pseudomonas from the results of morphological, cultural, and biochemical tests. The optimum conditions of medium for the growth and the degradation of TCHDPE were TCHDPE 500 ppm, (NH4)2SO4 0.1% as the nitrogen source, initial pH 7.0±0.1, and 37℃, respectively. In this conditions, the regradation rate of TCHDPE was about 97%. Pseudomonas sp. EL-O47R was tested for resistance to several metal compounds and antibiotics. Pseudomonas sp. EL-O47R was moderately grown to Cd(NO3)2, ZnCl2, AgSO4, CuSO4 and HgCl2. This strain was sensitive to rifampicin and kanamycln but resistant to ampicillin, penicillin, tetracyclin and chloramphenlcol. Pseudomonas sp. EL-O47R was grown structurally related com- pounds and potential metabolites of TCHDPE, and has the stability on TCHDPE biodegradation.

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Biodegradation Kinetics of Phenol and pcresol by Micrococcus sp. M1 (Micrococcus sp. M1에 의한 Phenol과 p-Creso의 생분해 Kinetics)

  • Son, Hong-Joo;Jang, Woong-Seok;Lee, Geon;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.6 no.2
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    • pp.153-163
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    • 1997
  • In order to fad the most fitted biodegradation model, biodegradation kinetics model to the initial phenol and p-cresot concentrations were investigated and had been fitted by the linear regression. Bacteria capable of degrading p-cresol were isolated from soil by enrichment culture technique. Among them, strain Ml capable of degradillg p.rcresol has also degraded phenal and was identified as the genus Micrococcus from the results from of taxonomical studies. The optimal tonditlons for the biodegradation of phenal and p-cresol by Micrococcus sp. Ml were $NH_4NO_3$ 0.05%, pH 7.0, 3$0^{\circ}C$, respectively, and medium volume 100m1/250m1 shaking flask. iwicrococcus sp. Ml was able to grow on phenal concentration up to 14mM and p-cresol concelltration up to 0.8mM. With increasing substrate concentraction, the lag period increased, but the maximum specific growth rates decreased. The yield coefficient decreased with increasing substrate concentation. The biodegradation kinetics of phenol and p-cresol were best described by Monod with growth model for every experimented concentration. In cultivation of mixed substrate, p-cresol was degraded first and phenol was second. This result implies that p-cresol and phenol was not degraded simultaneously.

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