AN EFFICIENT GENERAL-PURPOSE CULTURE MEDIUM FOR AEROBES AND ANAEROBES

1966 ◽  
Vol 12 (2) ◽  
pp. 323-331 ◽  
Author(s):  
Omar Khairat
Keyword(s):  
Growth Factors ◽  
Culture Medium ◽  
Agar Medium ◽  
General Purpose ◽  
Strictly Anaerobic ◽  
Viridans Streptococci ◽  
Heat Stable ◽  
Agar Media ◽  
Microbial Strains ◽  
Stable Growth

Thirty-three microbial strains (all but one freshly isolated from human pathologic material) and comprising aerobes, anaerobes, and CO2-dependent bacteria, were used to compare the growth capabilities of a new general-purpose medium, with two commonly used media; a heart infusion and a soy agar.The author"s new medium proved superior since it grew organisms (for example a pneumococcus) that failed to grow, even from massive inocula, on the two other media. A strictly anaerobic streptococcus from a bacteremia grew aerobically on the surface of the author"s medium while massive inocula of the same anaerobe failed to grow even anaerobically on the two other agar media. When colony size was compared (2070 colonies were measured) the author"s medium proved superior for 29 of the33 organisms tested.The soy agar medium gave poor α-hemolysis with pneumococci and viridans streptococci when compared with the two other media. The synergistic effect of both liver digest and yeast extract used in the author"s medium, and the heat-stable growth factors derived from them, are believed to be responsible for its superiority as a general-purpose medium for both aerobes and anaerobes.

Assessment of different selective agar media for enumeration and isolation ofListeriafrom dairy products

1988 ◽  
Vol 55 (4) ◽  
pp. 579-583 ◽  
Author(s):  
Lucas Dominguez ◽  
José Francisco Fernández ◽  
Victor Briones ◽  
José Luis Blanco ◽  
Guillermo Suárez
Keyword(s):  
Dairy Products ◽  
Agar Medium ◽  
Raw Milk ◽  
Superior Performance ◽  
Direct Plating ◽  
Selective Agar ◽  
Natural Microflora ◽  
Agar Media ◽  
The Difference ◽  
Better Than

SummaryDifferent selective agar media were compared for the recovery and isolation of five species ofListeriafrom raw milk and cheese. The selective media examined were Beerens medium, MacBride medium and that described by Dominguezet al.(1984) with 6 mg/1 acriflavine, listeria selective agar medium (LSAM), and LSAM with 12 mg/1 acriflavine (LSAM × 2A); a non-selective yeast glucose Lemco agar was included for comparison. When the difference between listeria and the natural microflora of raw milk and cheese was 102cfu/ml, listeria could be isolated by direct plating on all media tested. When it was lower than 103–104cfu/ml, listeria were isolated by direct plating only on LSAM and LSAM × 2A. When the difference was greater than 104cfu/ml, a previous enrichment was necessary to isolate them. LSAM and LSAM × 2A media performed better than the other media tested for isolating listeria by direct plating and improved their isolation from dairy products. This superior performance was evaluated by the ability of these media to support colony formation of different species ofListeriatested, the easy recognition of these colonies from those formed by other microorganisms and by their capacity to inhibit the natural microflora of these foods.

scholarly journals Improvement of the Culture Medium for the Dichlorvos-Ammonia (DV-AM) Method to Selectively Detect Aflatoxigenic Fungi in Soil

Toxins ◽  
2018 ◽  
Vol 10 (12) ◽  
pp. 519 ◽  
Author(s):  
Kimiko Yabe ◽  
Haruna Ozaki ◽  
Takuya Maruyama ◽  
Keisuke Hayashi ◽  
Yuki Matto ◽  
...  
Keyword(s):  
Culture Medium ◽  
Agar Medium ◽  
Carbon Sources ◽  
Tween 80 ◽  
Selective Medium ◽  
Selective Detection ◽  
Aflatoxigenic Fungi ◽  
Aspergillus Nomius ◽  
Rhizopus Sp ◽  
Triton X

The dichlorvos-ammonia (DV-AM) method is a simple but sensitive visual method for detecting aflatoxigenic fungi. Here we sought to develop a selective medium that is appropriate for the growth of aflatoxigenic fungi among soil mycoflora. We examined the effects of different concentrations of carbon sources (sucrose and glucose) and detergents (deoxycholate (DOC), Triton X-100, and Tween 80) on microorganisms in soils, using agar medium supplemented with chloramphenicol. The results demonstrated that 5–10% sucrose concentrations and 0.1–0.15% DOC concentrations were appropriate for the selective detection of aflatoxigenic fungi in soil. We also identified the optimal constituents of the medium on which the normal rapid growth of Rhizopus sp. was completely inhibited. By using the new medium along with the DV-AM method, we succeeded in the isolation of aflatoxigenic fungi from non-agricultural fields in Fukui city, Japan. The fungi were identified as Aspergillus nomius based on their calmodulin gene sequences. These results indicate that the new medium will be useful in practice for the detection of aflatoxigenic fungi in soil samples including those from non-agricultural environments.

Refining insulin concentrations in culture medium containing growth factors BMP15 and GDF9: An in vitro study of the effects on follicle development of goats

2017 ◽  
Vol 185 ◽  
pp. 118-127 ◽  
Author(s):  
D.J. Dipaz-Berrocal ◽  
N.A.R. Sá ◽  
D.D. Guerreiro ◽  
J.J.H. Celestino ◽  
J. Leiva-Revilla ◽  
...  
Keyword(s):  
Growth Factors ◽  
Culture Medium ◽  
In Vitro Study ◽  
Vitro Study ◽  
Follicle Development

scholarly journals THE PRODUCTION OF SELECTIVE ADDITIVES TO GROWING MEDIUM DRIGALSKI LACTOSE AGAR

2019 ◽  
pp. 95-102
Author(s):  
Владимир Ермаков ◽  
Vlyadimir Ermakov ◽  
Оксана Датченко ◽  
Oksana Datchenko ◽  
Юлия Курлыкова ◽  
...  
Keyword(s):  
Nutrient Medium ◽  
First Generation ◽  
Animal Species ◽  
Agar Medium ◽  
Nutrient Media ◽  
Gram Negative ◽  
Selective Media ◽  
Growing Medium ◽  
Agar Media ◽  
Growth And Reproduction

The purpose of the study is to improve the selective supplement for selective media with the purpose to produce enterobacteria. Tasks of the study are to identify the sensitivity of strains obtained of enterobacteria in regard to an-tibiotics; develop a new selective supplement with antibiotics to the nutrient medium Drigalski Lactose Agar. Media should have a content that in the best way possible ensures the growth and reproduction of microorganisms of cer-tain species or family. Intensive biotechnology development and Microbiology allows today to develop new nutrient media and modify the already existing content of media. The object of the study was a new selective additive with antibiotics to the nutrient medium Drigalski Lactose Agar. 253 isolates of bacteria produced from the intestinal mi-crobiotope of different animal species have been the Material for research. The study was conducted in the period from 2010 to 2017. Carbenicillin 30±2.3 from the group of carboxypenicillins and piperacillin 37±2.5 from the group of ureidopenicillins, kanamycin 24±1.5, amikacin 26±1.7 and gentamicin 25±0,8, cefepime 38±3.2 from the group of IV generation cephalosporins, tetracycline 28±1.6, doxycycline 34±2.3 and chloramphenicol 31±2.5, nalidixic acid 37±2.8, trimethoprim 35±3,4 demonstrated the greatest antimicrobial activity against all cultures of enterobac-teria that has been achieved. The high resistance of enterobacteria was shown to benzylpenicillin from the group of natural penicillins, to streptomycin, cephalotine from the group of cephalosporins of the first generation, to polymyx-in B, to ofloxacin (tarivid) and metronidazole. Antibacterial drugs effective against the accompanying gram-positive and gram-negative microflora were considered as the samples of the selective components. Vancomycin from the group of glycopeptides, linezolid from the group of oxazolidinones, and telithromycin from the group of ketolides were chosen. Antibiotics vancomycin and telithromycin in a dose of 0.008 g/dm3, linezolid 0.004 g/dm3 were cho-sen as the selective additive to Drigalski Lactose Agar medium.

scholarly journals EXPRESSION OF b-XYLOSIDASE ENCODING GENE IN PHIS/ Bacillus megaterium MS SYSTEM

2015 ◽  
Vol 2 (1) ◽  
pp. 25 ◽  
Author(s):  
Sri Sumarsih
Keyword(s):  
Culture Medium ◽  
Culture Supernatant ◽  
Specific Activity ◽  
Agar Medium ◽  
Nitrilotriacetic Acid ◽  
Protoplast Transformation ◽  
E Coli ◽  
Recombinant Cells ◽  
Relative Molecular Weight ◽  
Encoding Gene

b-Xylosidase encoding gene from G. thermoleovorans IT-08 had been expressed in the pHIS1525/ B. megaterium MS941 system. The b-xylosidase gene (xyl) was inserted into plasmid pHIS1525 and propagated in E. coli DH10b. The recombinant plasmid was transformed into B. megaterium MS941 by protoplast transformation. Transformants were selected by growing the recombinant cells on solid LB medium containing tetracycline (10 µg/ ml). The expression of the b-xylosidase gene was assayed by overlaid the recombinant B. megaterium MS941 cell with agar medium containing 0.2% ethylumbelliferyl-b-D-xyloside (MUX). This research showed that the b-xylosidase gene was succesfully sub-cloned in pHIS1525 system and expressed by the recombinant B. megaterium MS941. Theaddition of 0.5% xylose into the culture medium could increase the activity of recombinantactivity of recombinant of recombinantb-xylosidase by 2.74 fold. The recombinant B. megaterium MS941 secreted 75.56% of the expressed b-xylosidase into culture medium. The crude extract b-xylosidase showed the optimum activity at 50° C and pH 6. The recombinant b-xylosidase was purified from culture supernatant by affinity chromatographic method using agarose containing Ni-NTA (Nickel-Nitrilotriacetic acid). The pure b-xylosidase showed a specific activity of 10.06 Unit/mg protein and relative molecular weight ± 58 kDa.

Basic FGF and TGF-beta 1 influence commitment to melanogenesis in neural crest-derived cells of avian embryos

Development ◽  
1991 ◽  
Vol 111 (2) ◽  
pp. 635-645 ◽  
Author(s):  
K.M. Stocker ◽  
L. Sherman ◽  
S. Rees ◽  
G. Ciment
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Neural Crest ◽  
Cell Fate ◽  
Transforming Growth Factor Beta ◽  
Culture Medium ◽  
Transforming Growth Factor ◽  
Neutralizing Antibody ◽  
Pigment Cells ◽  
Tgf Beta

In previous studies, we showed that neural crest (NC)-derived cells from embryonic quail dorsal root ganglia (DRG) and peripheral nerve (PN), which do not normally give rise to melanocytes, become committed to melanogenesis following treatment in culture with the phorbol ester drug 12-O-tetradecanoyl phorbol-13-acetate (TPA). These and other observations support the notion that melanocytes and Schwann cells are derived from a common bipotent intermediate in the neural crest lineage—the melanocyte/Schwann cell progenitor. In this study, we test the possibility that peptide growth factors found in the embryonic environment might act similarly to TPA to influence the fates of these cells. DRG and PN explants were cultured in medium supplemented with a variety of growth factors, and then the cultures were examined for the presence of pigment cells. We found that basic fibroblast growth factor (bFGF), but not various other growth factors, induced pigmentation in about 20% of these cultures. When low concentrations of TPA were included in the culture medium, bFGF augmented the TPA-induced pigmentation, significantly increasing the proportion of pigmented cultures. These effects of bFGF were age-dependent, and could be blocked by addition of a bFGF-neutralizing antibody to the culture medium. In contrast to these stimulatory effects of bFGF, transforming growth factor-beta 1 (TGF-beta 1) was found to inhibit the TPA- or bFGF-induced pigmentation of DRG cultures. These data suggest, therefore, that at least some NC-derived cells are responsive to bFGF and TGF-beta 1, and that these growth factors may play an important role in the control of NC cell fate.

Procedures for Recovery of Stressed and Injured Cells of Yersinia Enterocolitica from Meat and Meat Products

1989 ◽  
Vol 52 (5) ◽  
pp. 360-362 ◽  
Author(s):  
ZHEKO KOUNEV
Keyword(s):  
Yersinia Enterocolitica ◽  
Agar Medium ◽  
Alkali Treatment ◽  
Meat Products ◽  
The Other ◽  
Step Procedure ◽  
Selective Agar ◽  
Agar Media ◽  
Phosphate Buffered Saline

A new three-step procedure (TSP) for the recovery of Yersinia enterocolitica 0:3 from frozen meat, salted and dried meat products, raw dried meat products, and cooked perishable sausages, has been developed. The TSP is based mainly on enrichment in 0.15 M phosphate-buffered saline at 25°C. In the TSP, selected dilutions of samples are enriched for 1, 2, 3, 4, 24, and 48 h and then plated onto nonselective and selective agar media after or without alkali treatment. Additional enrichment was performed with half of the samples at 25°C for 24 h, and the rest at 4°C for up to 2 wk, followed by alkali treatment and plating on selective agar medium. Recovery of Y. enterocolitica was better using the TSP than the other method used for isolating the organism from meats.

Influence du mode de culture dans la toxinogénèse de Plectridium tetani

1970 ◽  
Vol 16 (2) ◽  
pp. 135-136 ◽  
Author(s):  
G. Vinet ◽  
V. Fredette
Keyword(s):  
Tetanus Toxin ◽  
Culture Medium ◽  
Strictly Anaerobic ◽  
Anaerobic Conditions

Production of tetanus toxin under strictly anaerobic conditions does not favor high titers comparable to those obtained when the surface of the culture medium is largely exposed to the air.

Low Incidence of Aeromonas sp. in Livestock Feces

1987 ◽  
Vol 50 (1) ◽  
pp. 66-69 ◽  
Author(s):  
NORMAN J. STERN ◽  
E. S. DRAZEK ◽  
S. W. JOSEPH
Keyword(s):  
Aeromonas Hydrophila ◽  
Recovery Rate ◽  
Agricultural Research ◽  
Agar Medium ◽  
Brain Heart Infusion ◽  
Aeromonas Sobria ◽  
Aeromonas Caviae ◽  
Retail Meat ◽  
Agar Media ◽  
Low Incidence

Pig, beef, sheep and turkey fecal specimens were assayed for recovery of inoculated Aeromonas sp. by directly plating the samples on five different agar media. Of these, starch-ampicillin was optimal with respect to selectivity and ability to differentiate from other resident microflora. Generally, the numbers of inoculated Aeromonas sp. recovered on starch-ampicillin agar were similar to those recovered on brain heart infusion and blood ampicillin agar media, and were 101 to 103 greater than the recovery rate on either MacConkey-ampicillin or cefsulodinirgasan-novobiocin agars. The sensitivity for the direct recovery of Aeromonas sp. from inoculated beef feces with naturally contaminating microflora, using streaked starch-ampicillin agar medium, was between 102 and 103 cells per gram. Using starch-ampicillin agar, the incidence of Aeromonas detected from feces of beef, pig, sheep and turkey held at the Beltsville Agricultural Research Center was one of 32, none of 22, none of 24 and three of 21, respectively. Based upon current taxonomic criteria, the isolate from the beef feces had characteristics consistent with both Aeromonas sobria and Aeromonas caviae, whereas three isolates from turkey feces were identified as A. caviae or Aeromonas hydrophila. The organism was isolated from five of five packages of ground beef from retail sources. The discrepancy in the consistent presence of the organism in retail meat suggests that many of the food isolates are probably not of fecal origin.

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.

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