Roux bottle

Streptomyces hygroscopicus NRRL 5491 was grown and maintained on oatmeal-tomato paste agar slants (T. G. Pridham et al. Antibiotic Annual 1956-1957, Medical Encyclopedia Inc., New York, p. 947) and in Roux bottles containing the same medium. Good growth was obtained after 7 days of incubation at 28°C. Spores from one Roux bottle were washed off and suspended into 50 ml of sterile distilled water. This suspension was used to inoculate the first stage inoculum. 

The basic need for a solid support guides all production choices involving industrial processes for adherent cells. A large variety of vessels has been developed for adherent cell cultures. Petri dishes, Roux bottles, T-flasks, and roller bottles are examples of cell culture vessels with a glass or polystyrene surface. The system of choice is dependent on the seal-ability of multiple steps, as well as the cost of equipment and qualified operators. 

Table 18.1 compares the relationship between cell culture surface area and bioreactor volume in many different culture systems usually used with adherent cells. For microcarriers, this coefficient might reach 60 cm2/ml of medium for culture area prepared with 10 mg of microcarriers per milliliter. For Roux bottles, this coefficient is around 3 cm2/ml. In cultures initiated with 2 mg of microcarriers per milliliter of medium, high cell densities of even 3 X 106 cells/ml are often reached, compared with smaller cell densities from 2 to 3 X 105 cells/ml usually observed in Roux bottle systems. Another great advantage of the use of microcarrier culture systems is the possibility of preparing cell cultures with hundreds or even thousands of liters (Montagnon et al., 1984). 

Some glass bottles, e.g. Roux bottles, are manufactured especially for growth of cells in culture and these have the advantage of improved optical qualities allowing easier microscopic examination of the growing cells. Some of the larger bottles are known by the name of the person who introduced them. 

Seed 8 X 106 cells into 90 mm Petri dishes. Seed 20-30 X 106 cells into Roux bottles. Only about half the seeded embryo cells attach and grow. If the cell concentration is lowered, the fraction which attaches decreases particularly at seedings of less than 1 X 106/90 mm dish (105 cells/ml)... 

Organism, medium, and growth conditions. Rhodopseudomonas sp. No.7 and its maintenance were described previously [2]. A basal salt medimn containing both 0.1% ethanol and 0.2% sodium bicarbonate or 0.27% sodium acetate was used for the main culture. The organism was grown at 30°C in screw-capped 1.5 liter Roux bottles filled with the medium under photoanaerobic conditions. 

The batch culture was carried out in a bioreactor (B. Braun) with a working volume of 11. Forty-eight hours after the last Roux bottle inoculation, the cells were inoculated at about 2 xlO viable cells H in the bioreactor. 

In SSF, several parameters such as particle size, moisture content, incubation time, initial pH, and the amount of nitrogen sources have been shown to be important factors affecting the fungal growth and chitosan production (Nwe and Stevens 2004). An important aspect related to the production of chitosan in SSF has been use of different kinds of laboratory model bioreactors including Erlenmeyer flasks, rotary drum bioreactors, roux bottles, trays, and glass columns to evaluate... 

Inoculate a Roux bottle from a stock slant and incubate for 24 hours at 32-35°C. Wash the growth into sterile distilled water and standardize the resulting suspension so that the 100 units per milliliter solution of polymyxin standard will produce an inhibition zone 15 mm. in diameter on seeded agar plates. 

Prepare a spore suspmision by growing the organism in Roux bottles on nutrient agar for one week at 37°C. Suspend the spores in sterile distilled water and heat for 30 minutes at 65°C. Wash the spore suspension three times with sterile distilled water. Heat again for 30 minutes at 65 °C. and resuspend in sterile distiUed water. Maintain the spore suspension at approximately 15°C. Determine by appropriate... 

A fresh slant (7-14 day old) of each strain was used in each replicate batch. The hay steep mycelial inoculum was prepared by blending each slant in sterile water and transferring 10 ml of the resulting suspension down both sides of one-liter Roux bottles containing 240 ml of sterile hay steep. Two bottles of each strain were inoculated per replication. Bottles were incubated at 25 C (room temperature) on laboratory counter tops. Bottles were incubated for at least two weeks or until the establishment of a confluent mat. 

The hay steep mats were harvested and used to inoculate sterile Czapek-Dox production medium. The mats were removed with sterile forceps and blended in 125 ml sterile water. Ten ml of this suspension were dispensed into sterile one-liter Roux bottles containing 240 ml of Czapek-Dox medium supplemented with 0.3% yeast extract. Bottles were incubated at 25°C (room temperature) on laboratory counter tops for two weeks. Individual mats were harvested, blotted dry and weighed. Each... 

Production of slaframine and swainsonine by five isolates of Rhizoctonia leguminicola was examined. Cultures were grown in one-liter Roux bottles on yeast extract-supplemented Czapek-Dox medium. Differences were noted in growth, slaframine, and swainsonine production between the various isolates and between replicate batches of the same isolate. In addition, the ratio of slaframine to swainsonine produced by each isolate varied. These data suggest that regulation of slaframine and swainsonine synthetic pathways may vary between strains of R. leguminicola and are undoubtedly subject to alteration by environmental and nutritional factors. 

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