Performance of Different Solid and Liquid Culture Media for the Improvement of Tuberculin Production

2020 ◽  
Keyword(s):  
Culture Media ◽  
Synthetic Medium ◽  
Chemical Properties ◽  
Economic Loss ◽  
Delayed Type Hypersensitivity ◽  
Basic Medium ◽  
Primary Methods ◽  
Lowenstein Jensen ◽  
Time Required ◽  
Modified Media

Tuberculosis (TB) is one of the most important zoonotic bacterial diseases. A huge economic loss which could be direct or indirect are associated with the disease. Currently, the primary methods used for detection of TB in humans and cattle include the measurement of a delayed type hypersensitivity to purified protein derivative (PPD). So, the need for preparation of purified PPD with adequate properties and increasing the final PPD yield with decreasing the time of tuberculin production has stimulated the interest in the development of its preparation. Our study was performed to compare between the standard and modified media for improving tuberculin production. Middle brook 7H10 agar medium was used as a modified basic medium for mycobacterial growth, followed by cultivation of mycobacteria on Middle brook 7H9 broth medium. For the production, strains were inoculated onto the culture medium (Dorest Henly synthetic medium). Other steps for tuberculin production was done according to standard Weighbridge protocol. The results demonstrated that the using of both Middle brook 7H10 agar and Middle brook 7H9 broth instead of Lowenstein-Jensen (LJ) and glycerin broth media which used in currently produced tuberculin, have better physical and chemical properties. In addition, reducing the time required for production by accelerating the time of microbial growth. Also, it was found that the tuberculin produced using modified media was slightly more potent or the same as currently tuberculin produced. So, both Middle brook 7H10 agar and Middle brook 7H9 broth media are recommended for production of tuberculin saving time and increasing potency of the product but more investigation was recommended for estimation types of protein present in both locally prepared and modified tuberculin.

scholarly journals Study on Isolation and Partial Purification of Lactase (β-galactosidase) Enzyme from Lactobacillus Bacteria Isolated from Yogurt

2011 ◽  
Vol 4 (1) ◽  
pp. 239 ◽  
Author(s):  
N. H. M. R. Mozumder ◽  
M. Akhtaruzzaman ◽  
M. A. Bakr ◽  
F. Tuj Zohra
Keyword(s):  
Specific Activity ◽  
Culture Media ◽  
Deae Cellulose ◽  
Chemical Properties ◽  
Total Activity ◽  
Partial Purification ◽  
Anion Exchange Column ◽  
Ammonium Sulphate Precipitation ◽  
Agar Media ◽  
Modified Media

Lactase has many applications in dairy industry including for the treatment of lactose intolerance. The present study was conducted to identify the activity of lactase enzyme produced by Lactobacillus bacteria isolated from yogurts available in Dhaka city. The strains were identified to be gram positive, catalase negative, fermentative and lactase producer when cultured on selective MRS agar media by using standard bacteriological procedures and techniques. The study revealed that enzymes produced by lactobacilli were capable to produce glucose from substrate lactose in lactose modified media using lactase assay Kit Glu IB and their highest protein concentration (17.25 mg/ml) was observed in the supernatant of culture media isolated from L. lactis. Highest total activity (850.69 U/l) and specific activity (50.04 U/mg) of lactase enzyme was observed in the strain of L. bulgaricus. The crude extract which showed highest activity was further purified by ammonium sulphate precipitation followed by anion exchange column chromatography (DEAE cellulose). Final specific activity and fold purification of lactase enzyme reached to 62.80 U/mg and 1.47 respectively. The highest physic-chemical properties (Effect of pH and temperature) of lactase enzyme were observed at PH 6.0 which was 43.98 U/mg of protein and at 70°c temperature which was 111.11 U/mg of protein.Keywords: β-Galactosidase; Specific activity; DEAE cellulose; Fold purification; Yogurt.© 2012 JSR Publications. ISSN: 2070-0237 (Print); 2070-0245 (Online). All rights reserved.doi: http://dx.doi.org/10.3329/jsr.v4i1.8478J. Sci. Res. 4 (1), 239-249 (2012)

PSEUDOURIDINE FROM CULTURE MEDIA OF AGROBACTERIUM TUMEFACIENS

1964 ◽  
Vol 10 (6) ◽  
pp. 867-875 ◽  
Author(s):  
T. Suzuki ◽  
R. M. Hochster
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Melting Point ◽  
Culture Media ◽  
Synthetic Medium ◽  
Periodate Oxidation ◽  
Chemical Properties ◽  
Crystalline Form ◽  
Infrared Spectrophotometry ◽  
Elementary Analysis ◽  
Color Reactions

Cultures of Agrobacterium tumefaciens grown on a synthetic medium containing glucose plus a pyrimidine produced a pyrimidine riboside with unusual chemical properties. This substance was isolated in crystalline form and identified as 5-ribosyluracil (pseudouridine) on the basis of data obtained from ion exchange and paper chromatography, electrophoresis, ultraviolet and infrared spectrophotometry, color reactions, melting point, elementary analysis, hydrazinolysis, and periodate oxidation followed by bromine treatment.

scholarly journals Comparison of four culture media for isolation of Mycobacterium avium complex from porcine tissues

1979 ◽  
Vol 9 (2) ◽  
pp. 194-196
Author(s):  
C O Thoen ◽  
E M Himes ◽  
J L Jarnagin ◽  
R Harrington
Keyword(s):  
Mycobacterium Avium ◽  
Mycobacterium Avium Complex ◽  
Culture Media ◽  
Agar Medium ◽  
Egg Yolk ◽  
Sodium Pyruvate ◽  
Acid Fast Bacilli ◽  
Lowenstein Jensen ◽  
Time Required ◽  
Porcine Tissues

The efficiency of four culture media was compared for the isolation of Mycobacterium avium complex from 197 procine tissues. In 82 tissues with microscopic granulomas and acid-fast bacilli, a significantly greater number of isolates were obtained on Middlebrook 7H10 medium with sodium pyruvate than on Stonebrink medium, Herrold egg yolk agar medium, or Lowenstein-Jensen medium (P=0.01). In 46 tissues in which no microscopic granulomas or acid-fast bacilli were observed, a significantly greater number of isolates were made on Middlebrook 7H10 medium or Herrold egg yolk agar medium than on Stonebrink medium or on Lowenstein-Jensen medium (P=0.01). The time required to grow M. avium complex on Lowenstein-Jensen medium was significantly greater than the time required to observe growth on Stonebrink, Middlebrook 7H10, or Herrold egg yolk agar medium (p=0.001).

scholarly journals Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable Mycobacterium tuberculosis

2021 ◽  
Vol 10 (15) ◽  
pp. 3249
Author(s):  
Annelies W. Mesman ◽  
Seung-Hun Baek ◽  
Chuan-Chin Huang ◽  
Young-Mi Kim ◽  
Sang-Nae Cho ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Media ◽  
Multidrug Resistant ◽  
Culture Conditions ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Solid Culture ◽  
Lowenstein Jensen ◽  
Genetic Mechanisms ◽  
Lipid Resuscitation

An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable Mycobacterium tuberculosis (Mtb) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom Mtb could not be grown in standard solid culture media. When we attempted to re-grow Mtb from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of Mtb from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with Mtb’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance.

scholarly journals BCG Substrains Change Their Outermost Surface as a Function of Growth Media

Vaccines ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 40
Author(s):  
Sandra Guallar-Garrido ◽  
Farners Almiñana-Rapún ◽  
Víctor Campo-Pérez ◽  
Eduard Torrents ◽  
Marina Luquin ◽  
...  
Keyword(s):  
Culture Media ◽  
Chemical Properties ◽  
Growth Conditions ◽  
Neutral Red ◽  
Culture Conditions ◽  
Host Cells ◽  
Growth Media ◽  
Phenotypic Characteristics ◽  
Outermost Surface ◽  
Red Dye

Mycobacterium bovis bacillus Calmette-Guérin (BCG) efficacy as an immunotherapy tool can be influenced by the genetic background or immune status of the treated population and by the BCG substrain used. BCG comprises several substrains with genetic differences that elicit diverse phenotypic characteristics. Moreover, modifications of phenotypic characteristics can be influenced by culture conditions. However, several culture media formulations are used worldwide to produce BCG. To elucidate the influence of growth conditions on BCG characteristics, five different substrains were grown on two culture media, and the lipidic profile and physico-chemical properties were evaluated. Our results show that each BCG substrain displays a variety of lipidic profiles on the outermost surface depending on the growth conditions. These modifications lead to a breadth of hydrophobicity patterns and a different ability to reduce neutral red dye within the same BCG substrain, suggesting the influence of BCG growth conditions on the interaction between BCG cells and host cells.

scholarly journals 3D Printed Laminated CaCO3-Nanocellulose Films as Controlled-Release 5-Fluorouracil

Polymers ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 986 ◽  
Author(s):  
Denesh Mohan ◽  
Nur Fatin Khairullah ◽  
Yan Ping How ◽  
Mohd Shaiful Sajab ◽  
Hatika Kaco
Keyword(s):  
Drug Delivery ◽  
Chemical Properties ◽  
Pharmaceutical Compounds ◽  
The Body ◽  
Therapeutic Effects ◽  
Layer By Layer ◽  
3D Printed ◽  
Time Required ◽  
Uptake And Release ◽  
And Chemical Properties

Drug delivery constitutes the formulations, technologies, and systems for the transport of pharmaceutical compounds to specific areas in the body to exert safe therapeutic effects. The main criteria for selecting the correct medium for drug delivery are the quantity of the drug being carried and the amount of time required to release the drug. Hence, this research aimed to improve the aforementioned criteria by synthesizing a medium based on calcium carbonate-nanocellulose composite and evaluating its efficiency as a medium for drug delivery. Specifically, the efficiency was assessed in terms of the rates of uptake and release of 5-fluorouracil. Through the evaluation of the morphological and chemical properties of the synthesized composite, the established 3D printing profiles of nanocellulose and CaCO3 took place following the layer-by-layer films. The 3D printed double laminated CaCO3-nanocellulose managed to release the 5-fluorouracil as an effective single composition and in a time-controlled manner.

80 DEVELOPMENT OF A SYNTHETIC MEDIUM FOR THE IN VITRO CULTURE OF BOVINE EMBRYOS

2014 ◽  
Vol 26 (1) ◽  
pp. 154 ◽  
Author(s):  
D. Moreno ◽  
A. Neira ◽  
L. Dubreil ◽  
L. Liegeois ◽  
S. Destrumelle ◽  
...  
Keyword(s):  
Growth Factors ◽  
Embryo Culture ◽  
Disease Transmission ◽  
Culture Medium ◽  
Culture Media ◽  
Synthetic Medium ◽  
Wilcoxon Test ◽  
Blastocyst Development ◽  
Negative Effect

In the majority of media for embryo culture, 2 of typical components used are FCS or BSA; however, the presence of FCS in the culture medium has been shown to have a negative effect on embryo quality and the use of animal-derived proteins in culture media increases the risks of disease transmission through in vitro embryo production. The aim of this study was to develop an in vitro embryo culture medium free from FCS and BSA, but with the addition of various growth factors and cytokines (GF-CYK: IGF-I, IGF-II, bFGF, LIF, GM-CSF) 50 ng mL–1 and (TGF-β1) 100 ng mL–1 supplemented with hyaluronan (HA) and recombinant albumin (RA). Bovine oocytes (n = 1043, 6 replicates) from abattoir ovaries were matured in TCM-199 medium with 60 μg mL–1 penicillin, 60 μg mL–1 streptomycin, and 10 ng mL–1 EGF for 24 h at 39°C and 5% CO2 in humidified air. Afterward, the oocytes were fertilized in IVF-TALP medium with 6 mg mL–1 fatty acid-free BSA and 1.7 IU mL–1 heparin for 18 h under the same conditions. After fertilization, presumptive zygotes were divided into two groups and cultured in 30 μL droplets of SOF supplemented with (1) 0.4% BSA + 5 μg mL–1 insulin, 5 μg mL–1 transferrin, and 5 ng mL–1 selenium (ITS) as a control; or (2) GF-CYK + 0.5 mg mL–1 HA + 0.15% RA (M1). Droplets were preserved under mineral oil in a humidified atmosphere of 5% CO2, 5% O2, and 90% N2 at 39°C. Blastocyst development and blastocyst diameter was observed at 7 and 8 days post-fertilization (dpf). Developmental and diameter data were analysed using the Wilcoxon test by using R software. The blastocyst rates were not significantly different between the control and M1 medium: at 7 dpf (22.9% ± 4.8 and 30.2% ± 3.0), and at 8 dpf (29.6% ± 5.1 and 37.4% ± 2.0 respectively; P > 0.05). The blastocyst diameter obtained with the M1 medium was significantly greater (P < 0.05) than that of the control at 7 dpf (173.3 μm ± 4.9 and 157.2 μm ± 4.1, respectively); however, no significant differences were observed at 8 dpf (190.3 μm ± 5.2 and 179.7 μm ± 5.3, respectively). In conclusion, the FCS- and BSA-free medium with GF-CYK, HA, and RA (M1) showed a comparable development rate to the control medium at 7 and 8 dpf. These growth factors and cytokines in association with hyaluronan and recombinant albumin have a synergistic action by promoting an increase in the blastocyst diameter at 7 dpf. This is fully synthetic method of embryo culture; it presents a valuable tool to reduce the risks of disease transmission via embryo transfer.

scholarly journals Nanoparticle colloidal stability in cell culture media and impact on cellular interactions

2015 ◽  
Vol 44 (17) ◽  
pp. 6287-6305 ◽  
Author(s):  
Thomas L. Moore ◽  
Laura Rodriguez-Lorenzo ◽  
Vera Hirsch ◽  
Sandor Balog ◽  
Dominic Urban ◽  
...  
Keyword(s):  
Cell Culture ◽  
Colloidal Stability ◽  
Culture Media ◽  
Chemical Properties ◽  
Cellular Interactions ◽  
Biological Media ◽  
Cell Culture Media ◽  
Biological Fate ◽  
Physico Chemical ◽  
Shed Light

This review discusses nanoparticle colloidal stability in biological media in an attempt to shed light on the difficulty correlating nanoparticle physico-chemical properties and biological fate.

scholarly journals Comparison of Different Microscopic Methods with Conventional TB Culture

1970 ◽  
Vol 1 (1) ◽  
pp. 46-50 ◽  
Author(s):  
Farjana Rahman ◽  
Saurab Kishore Munshi ◽  
SM Mostofa Kamal ◽  
ASM Matiur Rahman ◽  
M Majibur Rahman ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Culture Media ◽  
Light Emitting Diode ◽  
Empirical Therapy ◽  
Culture Method ◽  
Resistance Rate ◽  
Clinical Samples ◽  
Mdr Tb ◽  
Directly Observed Treatment ◽  
Lowenstein Jensen

Tuberculosis is considered as the greatest cause of death worldwide. In the developing countries including Bangladesh, population density, poverty, malnutrition, and highly congested environment may create substantial risk for infection with Mycobacterium tuberculosis. The principal obstacle in the treatment of tuberculosis is the consumption of time and inaccurate diagnosis as well. Therefore, a retrospective study was carried out in order to compare results obtained by both the conventional AFB (Acid Fast Bacilli) microscopy and Lowenstein-Jensen (L-J) culture method for detection of Mycobacterium spp. in clinical samples from different categories of patients. Among one hundred and fifty samples, 83 (55.3%) AFB+ results were found under Bright-Field (BF) microscopy, 78 (52%) AFB+ and 91 (60.7%) AFB+ results were observed under conventional and Light Emitting Diode (LED) fluorescence microscopy. On L-J culture media, 103 (68.7%) AFB+ isolates were found which reveals that the culture could be a gold standard for diagnosis of TB. Significantly, the sensitivity and specificity of LED fluorescence microscopy was higher than those of other microscopic methods compared to culture. The resistance rate of isolates was higher for isoniazid (65%) and rifampicin (63.1%) than streptomycin (53.4%) and ethambutanol (38.8%). The isolates showed resistance to more than one drug. Among one hundred and three cases, total 36 (24.7%) cases reported as MDR-TB (resistant to all first-line drugs) in previously treated patients but it was nil in the new cases. Thus, this study emphasizes the need of Directly Observed Treatment Short Course (DOTS) program for selection of proper anti-tubercular drug therapy towards the patient rather than providing them with empirical therapy and thereby reducing the alarming increase of drug resistance. DOI: http://dx.doi.org/10.3329/sjm.v1i1.9133 Stamford Journal of Microbiology, Vol.1(1), July 2011, p.46-50

scholarly journals The Role of Smooth Muscle Cells in Vessel Wall Pathophysiology and Reconstruction Using Bioactive Synthetic Polymers

2011 ◽  
pp. 419-437 ◽  
Author(s):  
M. PAŘÍZEK ◽  
K. NOVOTNÁ ◽  
L. BAČÁKOVÁ
Keyword(s):  
Cell Culture ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Tissue ◽  
Culture Media ◽  
Vascular Grafts ◽  
Chemical Properties ◽  
Vascular Diseases ◽  
Cyclic Strain ◽  
Muscle Cells

This review summarizes recent trends in the construction of bioartificial vascular replacements, i.e. hybrid grafts containing synthetic polymeric scaffolds and cells. In these advanced replacements, vascular smooth muscle cells (VSMC) should be considered as a physiological component, although it is known that activation of the migration and proliferation of VSMC plays an important role in the onset and development of vascular diseases, and also in restenosis of currently used vascular grafts. Therefore, in novel bioartificial vascular grafts, VSMCs should be kept in quiescent mature contractile phenotype. This can be achieved by (1) appropriate physical and chemical properties of the material, such as its chemical composition, polarity, wettability, surface roughness and topography, electrical charge and conductivity, functionalization with biomolecules and mechanical properties, (2) appropriate cell culture conditions, such as composition of cell culture media and dynamic load, namely cyclic strain, and (3) the presence of a confluent, mature, semipermeable, non-thrombogenic and non-immunogenic endothelial cell (EC) barrier, covering the luminal surface of the graft and separating the VSMCs from the blood. Both VSMCs and ECs can also be differentiated from stem and progenitor cells of various sources. In the case of degradable scaffolds, the material will gradually be removed by the cells and will be replaced by their own new extracellular matrix. Thus, the material component in advanced blood vessel substitutes acts as a temporary scaffold that promotes regeneration of the damaged vascular tissue.

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