lishment unless the virus has been prepared and handled in accordance with the provisions of Parts 101 to 122 of this subchapter.

§ 118.12 Filling and labeling containers.

No immediate or true container of hogcholera virus shall be filled in whole or in part, and no label shall be affixed to such container, except under the supervision of an inspector.

HYPERIMMUNIZING VIRUS

§ 118.25 Inoculations for hyperimmunizing virus.

For use in the production of hyperimmunizing virus, licensees shall inoculate healthy young pigs weighing not more than 160 pounds each with at least 2 cc. of highly virulent hog-cholera virus: Provided, That when hog cholera from pen infection is manifested by the animals after the fourth day subsequent to admission to the premises of the licensed establishment, they need not be so inoculated.

§ 118.26 Requirements for hyperimmunizing virus.

Hyperimmunizing virus shall be collected at licensed establishments only from pigs which are observed on veterinary inspection to be visibly sick with hog cholera and which manifest well-marked and increasingly grave symptoms thereof attended with progressively abnormal temperatures common to the acute type of this disease.

SIMULTANEOUS VIRUS

§ 118.30 Inoculations for simultaneous virus.

(a) For use in the production of simultaneous virus, licensees shall inoculate young healthy pigs of good quality with at least 2 cc. each of highly virulent virus. Such pigs when inoculated shall weigh not less than 40 pounds nor more than 125 pounds.

(b) Pigs which are eligible only for the production of hyperimmunizing virus shall be inoculated and held in separate pens from those to be used for simultaneous virus. Such separation shall be made on or before the third day after inoculation and such pigs held thereafter in separate pens until released by the inspector.

pigs which become visibly sick on or before the third day, or subsequent to the seventh day after the time of inoculation. The physical condition of all pigs from which simultaneous virus is to be collected shall be recorded daily on and after the third day subsequent to inoculation. The observations required by the regulations in this part to be made on the third day may be made on the fourth day if the third day falls on Sunday or a holiday.

§ 118.32

Requirements for simultaneous virus, etc.

(a) Simultaneous virus and other hog-cholera virus intended for the inoculation of pigs for any purpose shall be collected at licensed establishments only from pigs which are observed by an inspector to be visibly sick with hog cholera within 7 days after the time of inoculation and which manifest wellmarked and increasingly grave symptoms of hog cholera attended with proigressively abnormal temperatures common to the acute type of this disease.

(b) Simultaneous virus shall be prepared in licensed establishments in batches of not to exceed 50,000 cc. The defibrinated blood in each batch shall not exceed 45,000 cc. and shall be mixed thoroughly in a single container before phenolization. All simultaneous virus shall be constantly agitated during the bottling operation.

§ 118.33

Samples of simultaneous virus.

The following representative samples of simultaneous virus shall be taken at licensed establishments and properly identified by an inspector: (a) At time of mixing but before phenolization, (1) "purity test sample" of not less than 30 cc. in a single container, (2) "test sample A❞ of not less than 5 cc. in a single container; (b) After mixing and phenolization, (1) "phenol test sample" of not less than 30 cc. in one container, (2) one reserve sample of 30 cc. to be forwarded to the Division in event the pigeon or mouse test is unsatisfactory and to be returned to the licensee if tests of the sample are satisfactory, (3) "test sample B" of not less than 5 cc. in a single container; (c) At time of bottling, a "stock sample" of at least 30 cc. in one container. All "A" and "B" test samples shall be held at approximately 75° F. under Division lock until used. All other samples shall be held under Division lock at 35° to 45° F.

§ 118.36

Disposition of samples of simultaneous virus.

At least one container of the stock sample of simultaneous virus shall be held at the licensed establishment unopened in the manner provided in § 102.77 (c) of this subchapter for at least 3 months after the latest expiration date shown upon the labels affixed to the immediate or true containers of the product corresponding to the sample.

§ 118.37 Test animals.

Two healthy calves, with mouths free from abrasions, as described in § 117.3 of this subchapter, or three healthy pigs immunized by the simultaneous treatment against hog cholera for at least 14 days, shall be furnished for intravenous injection with the purity test sample. These animals shall be given veterinary inspection immediately before the test is begun. All himals used for the testing of simultaneous virus shall be injected only under the supervision of an inspector and shall be marked as provided in Parts 101 to 122 of this subchapter. All test animals shall be given veterinary inspection as frequently as practicable during the test period to determine whether any symptoms or lesions of a vesicular or other disease develop.

§ 118.38 Purity test of simultaneous virus.

Each of the animals selected for testing the purity of simultaneous virus at licensed establishments shall be injected with 15 cc. of the purity-test sample into either the auricular or the jugular vein

If none of the animals which are treated with hog-cholera virus as prescribed in § 118.38 manifests symptoms of any infectious, contagious, or communicable disease, or if only one animal develops hog cholera, the test will be declared "satisfactory for purity," and the product released for marketing: Provided, It is otherwise satisfactory under the provisions of the regulations. Should any of the animals in the test succumb or should more than one develop hog cholera, another test may be made as in the first instance, except that not less than 15 cc. of the phenolized virus shall be used for the inoculation of each animal.

§ 118.41 Swine erysipelas.

Representative samples of each batch or serial of simultaneous virus shall be tested at licensed establishments in the following manner to determine its freedom from swine erysipelas (Erysipelothrix rhusiopathiae):

(a) Within 1 day after the first virus in a batch is collected, at least 1 cc. of test sample A shall be injected intramuscularly into each of three or more young pigeons or 0.2 cc. of such sample shall be injected subcutaneously into each of three or more suitable mice susceptible to swine erysipelas. These test animals and birds shall be held under the observation of an inspector for 10 or more days after being injected with the virus under test.

(b) Three or more days after phenolization of the batch of virus, at least 1 cc. of test sample B shall be injected intra-muscularly into each of three or more young pigeons or 0.2 cc. of such sample shall be injected subcutaneously into each of three or more suitable mice susceptible to swine erysipelas. These test animals and birds shall be held

under the observation of an inspector for 7 or more days after being injected with the virus under test.

(c) If all test animals or birds injected with test sample A survive for 10 days or more, and all test animals or birds injected with test sample B survive for 7 days or more, after injection, the batch or serial represented by the samples may be marketed if it otherwise conforms to the requirements of Parts 101 to 122 of this subchapter.

(d) Should any of the inoculated animals or birds die during the test, the product shall not be released for marketing and the reserve 30-cc. sample shall be forwarded to the Division.

(e) All animals or birds, after being once used in the tests provided in this section, shall be killed and their carcasses destroyed by incineration or tanking as provided in § 108.16 of this subchapter. Also all virus blood and simultaneous virus which are contaminated with erysipelothrix rhusiopathiae shall be destroyed in like manner. § 118.42 Marking "U.S. Released."

Each immediate or true container of simultaneous hog-cholera virus produced at licensed establishments which has been tested and found not to be worthless, contaminated, dangerous, or harmful, may have a cap affixed which, if approved by the Director, may bear the words "U. S. Released." These caps shall be affixed to the aforesaid containers only under the supervision of an inspector and shall be held under Division lock except when needed for this purpose. No simultaneous virus shall be released for marketing unless and until all information required by the regulations has been affixed to the containers thereof under supervision of an inspector. All simultaneous virus on which the expiration date has expired shall be destroyed as prescribed in § 108.16 of this subchapter. § 118.43 Expiration date.

The expiration date placed on the label of each immediate or true container of simultaneous virus produced at licensed establishments shall be one of the following:

(a) A date within 90 days after the first blood in the batch was collected: Provided, That the simultaneous virus is stored and marketed in containers acceptable to the Division;

(b) A date within 120 days after the first blood in the batch was collected

ments shall be healthy at the time of hyperimmunization, and this fact shall be determined by a thorough veterinary inspection. The weight of each animal in a given group shall be determined and recorded accurately by the licensee before hyperimmunization of the group. § 119.5 Dosage of virus.

All hogs which are used to produce anti-hog-cholera serum at licensed establishments shall receive, for hyperimmunization, a single intravenous injection of at least 5 cc. of hog-cholera virus for each pound of the animal's weight when injected.

§ 119.6 Temperatures before bleeding.

The temperatures of the hogs in each group or lot used to produce anti-hogcholera serum at licensed establishments shall be determined under normal handling conditions and recorded accurately by the licensee either on the afternoon before, or on the day of, bleeding and at such other times as the inspector may require. There shall be provided clean, light quarters equipped with a satisfactory chute and all other facilities for expediting temperature taking and veterinary inspection.

§ 119.7 Inspection before bleeding.

All hogs which are used to produce anti-hog-cholera serum at licensed establishments shall be subjected to a thorough veterinary inspection before each bleeding. Groups containing any hogs that are lame or otherwise suspected of being affected with a vesicular disease shall be given special examination for vesicles and the like after thorough cleansing of their feet, including examination of the coronary bands, snouts, and lips. Only those hogs which are found to have a temperature of less than 104° F. and are free from any infectious, contagious, or communicable diseases or other abnormal conditions shall be bled for serum. No hyperimmune hog in a lot or group of like origin having a significant number of high temperatures or showing other abnormalities indicative of an infectious or communicable disease shall be subjected to bleeding until such conditions of the lot or group as a whole no longer exist. § 119.8 Bleeding and examination.

(a) Anti-hog-cholera serum shall be derived at licensed establishments only from hyperimmune hogs which have been subjected to not more than four

successive bleedings, except that additional bleedngs may be authorized by the Director in emergencies. The first bleeding shall take place not earlier than the eleventh day after hyperimmunization; subsequent bleedings shall not take place more frequently than once in 7 days; and the last bleeding shall be made on a date not later than 40 days after hyperimmunization: Provided, That, in emergencies, final bleeding may be deferred when specifically authorized by the Director.

(b) Autopsies shall be performed at licensed establishments on hyperimmune hogs that succumb in order to obtain, if possible, information as to the cause of death. Employees of the licensed establishment, under the supervision of an inspector, shall perform the labor incident to these examinations.

(c) Anti-hog-cholera serum derived at licensed establishments from final bleedings shall be kept separate from other serum until it has been determined by post mortem examination that the hog from which the serum is derived was not so affected with any infectious, contagious, or communicable disease or in such condition as to render the serum worthless, contaminated, dangerous, or harmful.

§ 119.9 Constitutional symptoms.

Anti-hog-cholera serum derived at licensed establishments from hogs which, after hyperimmunization, manifest symptoms indicative of an affection of a constitutional character other than those usually observed immediately following hyperimmunization shall not be mixed with other serum, unless after due consideration of the prevailing conditions, this action is permitted by a veterinary inspector. Such serum, if collected only from hogs as prescribed in § 119.8, may be prepared separately and tested as prescribed in Parts 101 to 122 of this subchapter and if, as a result of these tests, the product is found satisfactory, it may be marketed. Otherwise, the serum shall be destroyed as provided in § 108.16 of this subchapter under the supervision of an inspector.

§ 119.10 Post mortem examination.

(a) All hogs from which anti-hogcholera serum is derived at licensed establishments shall be subjected, after final bleeding, to a thorough post mortem examination by an inspector. If, as a result of such inspection, it is found

that any hog is so affected with any infectious, contagious, or communicable disease or is in such condition as to render the serum worthless, contaminated, dangerous, or harmful, the serum collected from such hog shall be destroyed by the licensee, as provided in § 108.16 of this chapter under the supervision of an inspector.

(b) If serum-producing hogs at a licensed establishment become exhausted as a result of tail bleeding, dressing of the animals may be permitted provided they are given veterinary inspection immediately before throat bleeding and provided the animals bleed properly. The carcasses of such hogs may be dressed for food if disposition thereof is made in accordance with the meat inspection regulations (Subchapter A of this chapter). The blood of such animals may be used for serum if the tail and throat bleeding operations are such that no more time elapses between tail bleeding and throat bleeding than is necessary for removing the animals from the tail-bleeding station and restraining them at a regular throat-bleeding station.

ANTI-HOG-CHOLERA SERUM PREPARATION PROCEDURE

§ 119.20 Heating; time and conditions. All anti-hog-cholera serum produced at licensed establishments shall be heated under the supervision of an inspector in such a manner as to subject the product and the entire container thereof to a temperature of 58.5° C. for 30 minutes with a tolerance of 0.5° above and below that temperature, by methods prescribed by the Director.

Metal containers of a capacity not to exceed 50 liters shall be used in heating anti-hog-cholera serum at licensed establishments. Such containers shall be equipped with satisfactory agitators, and facilities for cooling and preserving the product shall also be provided. All serum shall be handled prior to heating so that practically all "foam" is eliminated before beginning the heating process and shall be properly agitated while being heated, cooled, and preserved. Each container of serum at time of heating shall be so submerged that the water line in the bath will be at least 2 inches above the upper surface of the lid. No container or other equipment intended for heating, cooling, preserving, and storing