B. Detailed technology for the production of cattle whey in the slaughterhouses of Lyon. Method for obtaining blood serum of adult cattle for culturing animal and human cells 5 with bovine serum

Blood has already been drawn from 1,000 animals, the serum has been bottled and distributed free of charge to almost 20,000 children. Thus, it is shown that the industrial production of whey in a slaughterhouse in compliance with aseptic rules and sanitary regulations is possible.

The serotherapy enterprise does not provide additional guarantees - it does not have the right to perform an autopsy on the donor animal.

The technology used in our production may seem less strict from an aseptic point of view than the classical method. But it has a big advantage in speed, since the serum is completely produced on the day of blood collection.

If current circumstances have prompted us to produce the serum in a slaughterhouse, it is clear that this is a temporary measure, since hematogen and medicinal serums can only be produced in a specialized institute.

Selection of animals. In Lyon, Dr. Guier, the chief veterinarian of the slaughterhouse, and Dr. Fontenay, the veterinary inspector, themselves select donors from among the livestock intended to provide our city with meat. The selected animal is branded on the right shoulder to facilitate further control. After slaughter, the animals' organs are carefully checked. It is known that autopsy is the most reliable method of detecting tuberculosis.

Subsequent operations will show that the serum of the diseased animal was never consumed.

Taking blood from animals. In the blood collection room, the donor bull is securely held by an automatic yoke.

The veterinarian disinfects the skin with iodine at the level of the animal's neck and makes an incision in the area of ​​the jugular vein with a scalpel. Venous puncture is performed with a trocar sterilized by prolonged boiling. Once blood appears, an autoclaved rubber hose is attached to the trocar to connect it directly and aseptically to the defibrinator.

The defibrinating technology and method for sterilizing the defibrinator are described below.

From each animal, 8-10 liters of blood are obtained, which is weighed on scales located under the apparatus. To facilitate the sanitary control operation, each defibrinator has a label with the data of the donor animal.

It should be noted that blood sampling is carried out aseptically thanks to a closed circuit of all components: trocar, rubber hose and defibrinator, which are pre-sterilized.

A label with the date of blood collection accompanies the collected blood from the moment of venous puncture until it is converted into serum and stored in refrigerators.

Blood defibration. In most serotherapy establishments, blood poured into glass vessels separates the serum under the pressure of the load. Under these conditions, blood first obtained from cattle contains little serum (approximately 10%). Therefore, the serotherapy center in Lyon uses a special technology that produces 50% of the serum, and also in a shorter time.

Dr. Merrier was able to develop this method partly after the findings he made at the Royal Institute in Rotterdam and at the Serotherapeutic Institute in Milan.

Once blood is obtained from these institutes, it is defibrinated in sterile machines that resemble churns. Within 5 minutes, the blood is collected in a vessel protected from air. It is necessary to strictly observe the defibration time: if there is a lack of it, coagulation can occur, and if there is an excess, hemolysis (due to the rupture of red blood cells). It is necessary to use a laboratory clock that allows you to mark exactly 15 minutes of defibrination.

At the top of Fig. 4 shows that the number of blood draws has reached 1000, which is recorded on the Center’s emblem.

Centrifugation. Immediately after defibration, the devices are transferred to the laboratory, located a few meters from the blood collection room. The contents of each defibrinator are processed separately so that the serum of an animal that turns out to be sick can be removed.

Blood that does not clot after defibrination is passed through an Alfa Laval separator (a milk separator that we have adapted for the production of whey). Under the influence of centrifugal force, the blood is divided into equal parts: the red part contains red balls, and the transparent part makes up the serum (fibrin remains on the defibrinator blades).

From one animal, 8-10 liters of blood or approximately 4-5 liters of serum are obtained, which is poured into a Pyrex bottle, sterilized at 180°C. A defibrinator label is attached to the bottle, and a card is filled out under the same number to ensure sanitary control.

A special antiseptic is added to the serum, designed so that it is sufficiently active without disturbing the transparency and taste of the serum. For 1 liter of serum, also add 100 ml of a solution containing 1:1000 formaldehyde and 1:5000 syunuxol. (Between preparations, the centrifuge is thoroughly disinfected with an antiseptic.)

In principle, the red fraction of the blood should be returned to the butchers to make blood sausage, but most often it remains unused, so syrup can be prepared from it using the technology described below.

Special cases of preparing syrup from blood. The syrup has two advantages: it makes it possible to use the red part of the blood remaining after receiving the serum, and it has a pleasant taste that children like.

Due to the lack of glycerin, it is difficult to prepare syrup for long-term storage, but you can make a very active product using water and give it to children 2-3 tablespoons per day. 20% of drinking water is added to the red part of the blood and the syrup is stored in glaciers while sanitary control is carried out. Then an equal part of 100% sugar syrup is added (sugar can be obtained from the cards of the groups for whom the syrup is intended).

Lemon or orange extract is added to the syrup to neutralize the taste of blood and poured into 250 ml bottles.

Tyndallization of whey. Immediately after centrifugation, that is, less than an hour after blood collection, the serum is lysed at 56°C for an hour. To do this, it is lowered into a water bath with an automatically maintained temperature. Tyndallization at this temperature (at a higher temperature the whey coagulates) is necessary for partial sterilization of the whey, although the speed of its preparation in itself is a guarantee of asepsis.

It should be noted that each five-liter bottle is provided with the original defibrinator label, thus the numbering of the serum corresponds to the numbering of the donor animals.

Control card file. The card allows you to check at any time the origin of the donor animal, the stages of whey preparation, the bottling date, as well as the distribution of the whey.

Sanitary control. While the vessels are stored in the refrigeration chamber, veterinarian inspectors are engaged in sanitary control of donor animals. After their slaughter, a thorough autopsy is carried out to reveal the slightest symptoms of tuberculosis. If a disease is detected, the corresponding serum can be easily withdrawn. It is known that the blood of each animal is processed separately and the serum is contained in separate, numbered bottles.

Serotherapeutic center of the Public Committee of Childhood Hygiene at the Lyon Slaughterhouses

Control number...

Bovine whey in liters

The above card and label prevents any confusion.

In Lyon, sanitary measures are especially strict, as the same veterinarians select the animals, draw blood and check the meat.

Sterilization of equipment. Animal serum is an excellent breeding ground for microbes and can only be partially sterilized. At temperatures above 56° they coagulate and become cloudy if a strong antiseptic is added. Therefore, during all serum production operations, maximum sterility is required; equipment must be decontaminated before use.

Individual defibrinators are sterilized in the following way: the night before blood collection, they are filled with an antiseptic solution, and a few hours before blood collection, they are emptied of antiseptic using a tap located in the lower part of the device. Centrifuges used to produce serum are also treated with an antiseptic, including between processing the contents of each defibrinator.

All glassware, including five-liter vessels for storing whey, is sterilized in an electric oven at a temperature of 180°C.

250 ml bottles for serum are also sterilized at 180°C. To simplify the operation, the dishes are located in boxes, which are used during bottling and distribution to the public.

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COMPOSITION AND FORM OF RELEASE
Serum against pasteurellosis, salmonellosis, escherichiosis, parainfluenza-3 and infectious rhinotracheitis of cattle is a biological product obtained from the blood of oxen hyperimmunized with inactivated strains of the pathogens of salmonellosis, escherichiosis and pasteurellosis, avirulent strains of parainfluenza-3 and infectious rhinotracheitis viruses. In appearance, it is a light yellow liquid with a reddish tint; during storage, a slight sediment forms, which easily breaks when shaking the bottle. The serum is packaged in 100 ml bottles in dark glass bottles, hermetically sealed with polymer caps and rolled with aluminum caps.

PHARMACOLOGICAL PROPERTIES
Hyperimmune serum promotes the formation of passive immunity to the pathogens of salmonellosis, pasteurellosis, E. coli, infectious rhinotracheitis and parainfluenza-3 in cattle, the intensity of which is 10 days. To form intense active immunity in animals against infections, vaccines are used. The mechanism of action of the serum is based on the binding and neutralization of pathogenic antigens by specific antibodies of the biological product. Hyperimmune serum is effective for the treatment of the above diseases, especially in the early stages of the infectious process and goes well with drugs used for symptomatic treatment (antimicrobials and probiotics).

INDICATIONS
Prescribed to cattle for the purpose of specific prevention and treatment of salmonellosis, pasteurellosis, escherichiosis, parainfluenza-3 and infectious rhinotracheitis.

DOSES AND METHOD OF APPLICATION
Before administering the serum, syringes and needles must be sterilized. A separate sterile needle is used for each animal. Before use, the bottle must be shaken vigorously and the injection site treated with 70% ethyl alcohol. Before administration, the bottle should be heated in a water bath to a temperature of 36-38 ºС. For prophylactic purposes, the serum is administered to animals subcutaneously twice with an interval of 7-10 days: calves 20-30 ml, adults 30-60 ml. For therapeutic purposes, the serum is administered intramuscularly or intravenously in the initial stage of an infectious disease: 40-60 ml for calves, 60-120 ml for adults. The absence of a therapeutic effect after the injection indicates a different etiology of the disease and additional laboratory tests should be performed to clarify the diagnosis.

SIDE EFFECTS
In very rare cases, with increased individual sensitivity in animals, allergic reactions are possible.

CONTRAINDICATIONS
No contraindications have been identified when using the serum.

For cultural and bacteriological work in laboratory conditions, blood serum is of great importance, which is necessary both for the preparation of nutrient media for mammalian cell cultures, and as an integral part of microbiological nutrient media.

Cattle blood serum is widely used, since the raw materials for it are quite accessible in any quantity. However, horse serum, as well as blood serum from other animals, is no less in demand.

In the process of producing high-quality blood serum, great attention must be paid to blood separation and subsequent sterilization of the serum. Only with the use of the most modern filtration systems is maximum quality possible. However, to preserve the quality of the whey, it is important to use sterile containers. To seal, sterilized glass containers of various volumes (where serum from cattle and other animals is packaged) are sealed with rubber stoppers with aluminum caps.

Then each batch is assigned a number, which is duplicated in the product passport and on its label. However, blood serum does not reach the client immediately; before entering the market, they undergo thorough quality checks, and only after passing they receive quality certificates and permission to sell.

Horse serum, as well as bovine serum, can be stored for one year at a temperature of 2 to 8 degrees Celsius above zero. In a frozen state, the shelf life of the serum increases to 2 years.

We always have in stock bovine blood serum and other blood serums for use in laboratory practice. All our products (bovine serum, equine serum, fetal bovine serum or FBS) have quality certificates and registration certificates. We work with the best Russian and foreign serum manufacturers.

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Manufacturer: BioloT

Country Russia

Unit Meas.: bottle

Article: 1.1.1.3

Description

Bovine blood serum for cell cultures, liquid, sterile, in a 100 ml polyethylene bottle. It is a native serum obtained from the heart blood of cows up to three years old, by coagulation, centrifugation and sterilizing filtration through a system of membrane filters. May or may not contain preservatives

A component of nutrient media that stimulates the growth of human and animal cell cultures for the production of medical immunobiological drugs (MIBPs), as an additive in microbiological nutrient media. Used to detect toxigenic strains of Corynebacterium diphtheriae mitis with the formation of nonspecific precipitation bands. For use in medical, health care and research institutions

Transparent liquid from straw-yellow to reddish-orange. During storage, slight opalescence and slight flaky sediment may appear that breaks up when shaken.
Transparency (optical density units) - no more than 0.45;
pH value: from 7.4 to 8.2;
Protein content - in the range from 60.0 to 80.0 g/l;
Hemoglobin content - no more than 0.5 g/l;
Sterile, does not contain mycoplasmas and viruses;
Specific activity - for cell cultures of the LEC, LEK type or diploid cells of human embryonic skin-muscular tissue: formation of a monolayer for 3-4 days, proliferation index after the 5th passage of at least 2.0.
Release form: in polymer bottles, 100 ml, hermetically sealed.
Storage conditions: at a temperature of -10...20°C in a dark place. During defrosting, fibrin may appear, which does not affect the quality of the serum.
Shelf life: 1 year at temperatures from +2...8°C, 2 years at -10...-20°C. After the expiration date indicated on the packaging, it cannot be used.
Registered with Roszdravnadzor

INSTRUCTIONS

on the use of cattle blood serum

native nonspecific

(manufacturer organization: BiAgro Trading House LLC, Vladimir)

I. GENERAL INFORMATION

1. Native nonspecific bovine blood serum without preservative.

2. Native nonspecific cattle blood serum without preservative consists of native cattle blood serum.

3. In appearance, the serum is a transparent liquid of straw-yellow color, a slightly reddish tint is allowed; during storage, opalescence and a slight sediment are possible, which breaks up when shaken.

4. The serum is available in bottles: 200, 400, 450 cm3. The bottles are sealed with rubber stoppers and reinforced with aluminum caps.

A label is placed on each bottle indicating: the manufacturing organization, its address and trademark, name of the serum, quantity (cm3), batch number, date of manufacture (month, year), expiration date (month, year), state registration number, inscription “ For animals”, information on confirmation of conformity, storage conditions, service station designation.

The bottles are wrapped in wrapping paper and placed in cardboard boxes. Instructions for use are included in each box. A label is pasted onto the box indicating: the manufacturing organization, its address and trademark, the name of the serum, the designation of this service station, batch numbers, production date (month, year), expiration date (month, year), state registration number, storage conditions , information about confirmation of conformity, the number of bottles in the box, the inscription “For animals”.

5. Cattle blood serum is stored in a closed, dry place in boxes or transport containers at a temperature of 2? C to 10? C for one year. Storage at a temperature of minus 20? C for one year is allowed.

6. After opening, hermetically sealed vials are stored at a temperature from 2? C to 10? C for one year. Storage at a temperature of minus 20? C for one year is allowed.

7. If the integrity of the bottles is damaged, the presence of foreign impurities that do not break when shaking the flakes, changes in color, violation of capping and labeling, the serum must be disposed of by boiling or autoclaving at 0.11 MPa for 15 minutes.

8. Expired serum does not require special means for disposal.

II. BIOLOGICAL PROPERTIES

9. Cattle blood serum stimulates the growth of continuous, primary, hybrid, bacterial, fungal, mycoplasma cell cultures, as well as the reproduction of viruses.

III. ORDER OF APPLICATION

10. Cattle serum is used as a growth-stimulating additive to nutrient media used for growing cell cultures, bacteria, mycoplasmas, fungi, and virus reproduction.