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Collection of penicillin by submerged fermentation

The inoculum or “seed” for large fermentation tanks from 20,000 to 115,000 liters of capacity is prepared by the development of a stock culture of the fungus from spores that are dried on a substrate of nutrient agar. Several liters of culture medium, generally constitutes about 5 to 10% of the total, are prepared in a series of reservoirs and serve as seed for planting a large fermentation Cuba.

The four main phases of the production of penicillin are:

* Fermentation
* Separation of fermentation broth and mycelium extract of penicillin by means of solvents.
* Solvent Purification and training of the sodium salt of penicillin.
* Testing testing, storage and sale.

The breeding ground for fermentation is obtained by aqueous infusion of corn, adding a 2 to 3% lactose, and inorganic compounds are also added containing hydrogen, oxygen, phosphorus, sulfur, potassium, magnesium, nitrogen and traces of iron , copper and zinc. The addition of certain compounds that promote growth of the fungus should be avoided as they may be tolerated when administering the product, serious economic or disposal. After adjusting the pH to 4.5-5.0, the culture medium is passed to the fermenter, which is equipped with a vertical agitator, with an introduction of air system filter sterilized and coils to maintain the desired temperature. The fungus enters through sterile pipes using compressed air. During growth the medium is sterilized with steam under pressure, and temperature is kept between 23 and 25 º C. The sterile air allows aerobic fungal growth, and agitation facilitates uniform distribution within the liquid. It requires a volume of air per minute per volume of culture medium. The process is controlled intervals ranging between 3 and 6 hours after 50 to 90 hours growth is slowing, indicating that the fungus has been fully developed. The mass is cooled to 5 ° C. because of the instability of penicillin at room temperature, and separated the mycelium in a rotating drum filter.

In the old process, penicillin was extracted from the filtrate by adsorption on charcoal. It eluía amyl acetate, the eluate concentrated once cooled to 0 ° C and acidified to pH 2.0 with an organic acid. In the solvent extraction process is skipped the step of adsorption with active carbon and the filtrate (called “beer”) is adjusted to pH 2.5 with phosphoric acid in the same drive. It makes a continuous countercurrent extraction with amyl acetate and then with chloroform, concentrating on successive Podbielniak type centrifugal pumps, and the final liquid is treated with phosphate buffer and sodium bicarbonate to form the sodium salt. This product is sterilized by filtration and removes water and other asépticamente98 solvent crystallization, which is obtained crystalline penicillin, once dry can be packed in polythene bags or in glass or stainless steel.
Immunizing biologicals

Another area deals with bacterial vaccines, live vaccines and antitoxins. The immunity to a disease is accomplished by stimulating the formation of specific antibodies or by administration of antibodies previously formed.

This group includes vaccines for typhoid fever, plague, diphtheria, tetanus, influenza virus, mumps, polio, rabies, smallpox and typhus. The vaccine of influenza virus is unique, it is a mixture of two or more races, each of which is produced and tested separately, and that comes from cultures supplied by the National Institutes of Health. The virus comes in a blister properly prepared it is diluted 100,000 times in eggs inoculated before.

The actual cycle of production begins to give birth to first fertile eggs are examined under the light, then disinfected by applying a solution of iodine and finally drilled a small strawberry. The virus is introduced through this small hole is sealed with collodion and incubated at 37.2 º C for 48hs. It removes a circular section of the outside of the egg and extract the live virus allantoic fluid is separated from the egg fluid by centrifugation, so that the heavier viral particles are sedimented at the bottom of the centrifuge tube. Collect the virus and resuspended in saline solution. The live virus is inactivated by treatment with formalin at 40 ° C for 24h.