Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations 14357

required by paragraph (e) of this section,

are required for a serial of autogenous

biologic with 50 or fewer containers.

(9) Miscellaneous: The number of

samples from products not in the

categories provided for in paragraphs

(b)(1) through (b)(8) of this section shall

be prescribed in the filed Outline of

Production for the product.

(c) Prelicensing and Outline of

Production changes: Samples needed to

support a license application or a

change in the Outline of Production for

a licensed product shall be submitted

only upon request from the animal and

Plant Health Inspection Service. Except

for miscellaneous products specified in

paragraph (b)(9) of this section, the

number of such samples shall be at least

one and one-half times the number

prescribed for such product in

paragraph (b) of this section. Samples of

Master Seeds and Master Cell Stocks

with a minimum individual volume of

1 ml shall be submitted as follows:

(1) Ten samples of Bacterial Master

Seeds.

(2) Thirteen samples of viral Master

Seeds or nonviral Master Seeds

requiring cell culture propagation. For

Master Seeds isolated or passed in a cell

line different from the species of

intended use, an additional 2 samples

are required for each additional species.

For Master Seeds grown in cell culture

and intended for use in more than one

species, an additional 2 samples are

required for each additional species.

(3) Thirty-six samples of at least 1 ml

each or six samples of at least 1 ml each,

one sample of at least 20 ml, and one

sample of at least 10 ml of Master Cell

Stocks. In the case of Master Cell Stocks

which are persistently infected with a

virus, an additional four samples of at

least 1 ml each are required. If these

persistently infected cell stocks are

intended for use in more than one

species, an additional two samples of at

least 1 ml each are required for each

additional species.

(4) Four samples of the Master Cell

Stock + n (highest passage) cells.

(d) Sterile diluent: A sample of Sterile

Diluent shall accompany each sample of

product, other than Marek’s Disease

Vaccine, if such diluent is required to

rehydrate or dilute the product before

use. The volume of diluent shall be an

appropriate amount to rehydrate or

dilute the product. Samples of Sterile

Diluent prepared for use with Marek’s

Disease Vaccine shall be submitted

upon request from the Animal and Plant

Health Inspection Service.

(e) Reserve samples shall be selected

from each serial and subserial of

biological product. Such samples shall

be selected at random from final

containers of completed product by an

employee of the Department, of the

licensee, or of the permittee, as

designated by the administrator. Each

sample shall:

(1) Consist of 5 single-dose packages,

2 multiple-dose packages, or 2

diagnostic test kits, except that, in the

case of diagnostic test kits in which

final packaging consists of multiple

microtiter test plates or strips, a sample

may consist of a specified number of

test plates or strips along with all other

test reagents as prescribed in a filed

Outline of Production;

(2) Be adequate in quantity for

appropriate examination and testing;

(3) Be truly representative and in final

containers;

(4) Be held in a special compartment

set aside by the licensee or permittee for

holding these samples under

refrigeration at the storage temperature

recommended on the labels for 6

months after the expiration date stated

on the labels. The samples that are

stored in this manner shall be delivered

to the Animal and Plant Health

Inspection Service upon request.

(Approved by the Office of Management and

Budget under control number 0579–0013)

§ 113.309 [Amended]

3. In § 113.309, paragraph (c)(4), the

words ‘‘Veterinary Services’’ are

removed and the words ‘‘Animal and

Plant Health Inspection Service’’ are

added in their place.

Done in Washington, DC, this 13th day of

March 1995.

Terry L. Medley,

Acting Administrator, Animal and Plant

Health Inspection Service.

[FR Doc. 95–6649 Filed 3–16–95; 8:45 am]

Billing Code 3410–34–M

9 CFR Part 113

[Docket No. 92–132–2]

Viruses, Serums, Toxins, and

Analogous Products; Revision of

Standard Requirements

AGENCY: Animal and Plant Health

Inspection Service, USDA.

ACTION: Final rule.

SUMMARY: This rule amends the

Standard Requirements concerning Dog

Safety Testing; Canine Distemper

Vaccine, Killed Virus; Canine Hepatitis

Vaccine, Killed Virus; Canine

Adenovirus Type 2 Vaccine, Killed

Virus; Mink Enteritis Vaccine, Killed

Virus; Canine Hepatitis Vaccine, Live

Virus; Canine Adenovirus Type 2

Vaccine, Live Virus; and Canine

Distemper Vaccine, Live Virus. The

amendments are necessary because new

test methods and procedures have been

developed that can replace current test

requirements and increase the validity

of test results. The effect of the

amendments is to provide new test

methods and procedures and to relax

some of the restrictions currently in

effect. Also, the Standard Requirement

for Canine Distemper Vaccine (Ferret

Virulent) is removed because this

vaccine is no longer manufactured.

EFFECTIVE DATE: April 17, 1995.

FOR FURTHER INFORMATION CONTACT:

Dr. David A. Espeseth, Deputy Director,

Animal and Plant Health Inspection

Service, Biotechnology, Biologics, and

Environmental Protection, Veterinary

Biologics, 4700 River Road Unit 148,

Riverdale, MD 20737–1228, (301) 734–

8245.

SUPPLEMENTARY INFORMATION:

Background

The regulations in 9 CFR part 113

‘‘Standard Requirements’’, (referred to

below as the regulations) consist of test

methods, procedures, and criteria

established by the Animal and Plant

Health Inspection Service (APHIS) for

the evaluation of veterinary biological

products based upon their purity, safety,

potency, and efficacy. The Agency

periodically reviews the regulations and

amends test methods and procedures as

required to ensure that they are

consistent with current scientific

knowledge. On July 23, 1993, we

published in the Federal Register (see

58 FR 39467–39473, Docket No. 92–

132–1) a proposed rule to update the

regulations based upon current

scientific knowledge.

We solicited comments concerning

our proposal for a 60-day comment

period ending September 21, 1993. We

received four comments by that date.

One commenter fully supported the

proposal as written. Three commenters

suggested changes to certain sections

related to the Standard Requirements.

These comments are discussed below.

Two commenters suggested changes

to § 113.204. Both commenters

indicated that the portion of the

regulations dealing with the time(s) of

feces collection for virus detection

required clarification, and suggested

that feces collection at some point from

day 4 to 8 would be appropriate.

APHIS believes that the above

comments have merit. APHIS agrees

that feces collection early or late in the

collection period, or more than once, is

unnecessary. Therefore, APHIS has

revised the regulations in

§ 113.204(b)(2) to specify that feces are
14358 Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations

to be collected once from day 4 through

day 8.

One of the two commenters stated

that the term ‘‘virus isolation’’ should be

defined and that methods other than

that specified, ‘‘virus isolation and/or

fluorescent antibody examination,’’

should be allowed for the detection of

virus in feces. APHIS agrees that not

only ‘‘virus isolation’’ but the whole

phrase ‘‘virus isolation and/or

fluorescent antibody examination’’

needs clarification. Therefore, we have

revised the regulations in

§ 113.204(b)(2) to specify that cell

culture with fluorescent antibody

examination is the acceptable method of

virus detection. APHIS does not agree,

however, with the suggestion that other

methods of virus detection should be

specified in the regulations presently.

We believe that cell culture with

fluorescent antibody examination is the

most sensitive and specific method of

virus detection. Should APHIS become

aware of another method that is superior

to that indicated, it would consider

rulemaking to specify that method in

the regulations.

One of the two commenters also

stated that unvaccinated control mink in

immunogenicity studies should not be

considered susceptible to challenge if

the animals exhibit clinical signs but do

not shed virus. The second commenter

stated that the determination of virus

shedding in animals used for

immunogenicity studies should not be

required if four or five of the five

unvaccinated control mink exhibit

clinical signs. APHIS does not agree

with either commenter. We believe that

the absence of appropriate clinical signs

in vaccinated mink challenged with

virulent mink enteritis virus together

with clinical signs in unvaccinated

control mink after challenge is sufficient

evidence of the effectiveness of the

challenge. We also believe that an

effective vaccine against mink enteritis

should prevent virus shedding. No

change in the regulations is made in

response to these comments.

As a final comment on § 113.204, one

commenter criticized what the

commenter thought was a Standard

Assay Method (SAM) developed by the

National Veterinary Services

Laboratories for challenging mink with

mink enteritis virus. No such SAM has

been prepared. What has been prepared

is more appropriately termed a ‘‘bench

protocol.’’ Since the protocol was not

addressed in the proposed rulemaking,

no change to the regulations is made

based on this comment.

Comments on the three other

Standard Requirements included in the

proposed rule (§§ 113.40, 113.201, and

113.306) were received from another

commenter. Two of the comments

related to the route of canine distemper

virus challenge and the requirements for

satisfactory vaccine performance in a

repeat immunogenicity study. The

commenters requested that § 113.306

concerning live virus vaccines be

changed to specify an intranasal rather

than the traditional intracerebral route

of challenge. No amendments to the

route of challenge or repeat

immunogenicity requirements were

proposed in § 113.306. An intracerebral

challenge has been used successfully for

many years with the live virus vaccine.

It was the proposed amendments to

§ 113.201 that changed the route of

challenge from intranasal to

intracerebral for killed virus vaccines to

be consistent with that specified for live

virus vaccines in § 113.306. Since the

commenters focussed only on § 113.306

and that section is not being amended

as to route of challenge, no change to

the regulations is made in response to

these commenters.

The same commenter also claimed

that the proposal would result in the

overuse of Master Seed and suggested

that material obtained after five passages

of Master Seed be used instead. APHIS

disagrees with this comment. In

requiring that Master Seed be used, the

proposed change is consistent with

other regulations in part 113. We believe

that testing the Master Seed is necessary

for a satisfactory determination of its

use. No change to the regulations is

made in response to this commenter.

Based on the rationale set forth in the

proposed rule and in this document, we

are adopting the provisions of the

proposed rule as a final rule, with the

changes discussed in this document.

Executive Order 12866 and Regulatory

Flexibility Act

This proposed rule has been reviewed

under Executive Order 12866. The rule

has been determined to be not

significant for the purposes of Executive

Order 12866 and, therefore, has not

been reviewed by the Office of

Management and Budget.

This final rule revises the current

Standard Requirements for certain

vaccines. Sections 113.201 and 113.202

are amended to revise the potency test

performed on each serial of product so

that fewer dogs will be used and

serology will be used instead of virus

challenge. Both of these changes will

decrease the costs of production to the

manufacturer. In § 113.204, the potency

test in mink is changed to require that

virus shedding be examined. This

change should result in only a minimal

increase in cost (less than $100 per test)

to the manufacturer. Other changes to

the Standard Requirements generally

update the Standard Requirements to

reflect current scientific knowledge. We

do not expect any increase in cost,

except as noted above, to the 200

biologics manufacturers affected by this

rule. In most cases, we expect the

changes will actually decrease the costs

for the manufacturers.

Under these circumstances, the

Administrator of the Animal and Plant

Health Inspection Service has

determined that this action will not

have a significant economic impact on

a substantial number of small entities.

Executive Order 12372

This program/activity is listed in the

category of Federal Domestic Assistance

under No. 10.025 and is subject to

Executive Order 12372, which requires

intergovernmental consultation with

State and local officials. (See 7 CFR part

3015, subpart V.)

Paperwork Reduction Act

This rule contains no new

information collection or recordkeeping

requirements under the Paperwork

Reduction Act of 1980 (44 U.S.C. 3501

et seq.).

Executive Order 12778

This final rule has been reviewed

under Executive Order 12778, Civil

Justice Reform. This rule: (1) Preempts

all State and local laws and regulations

that are in conflict with this rule; (2) has

no retroactive effect; and (3) does not

require administrative proceedings

before parties may file suit in court

challenging this rule.

List of Subjects in 9 CFR Part 113

Animal biologics, Exports, Imports,

Reporting and recordkeeping

requirements.

Accordingly, 9 CFR part 113 is

amended as follows:

PART 113—STANDARD

REQUIREMENTS

1. The authority citation for part 113

continues to read as follows:

Authority: 21 U.S.C. 151–159; 7 CFR 2.17,

2.51, and 371.2(d).

2. Section 113.40 is revised to read as

follows:

§ 113.40 Dog safety tests.

The safety tests provided in this

section shall be conducted when

prescribed in a Standard Requirement or

in the filed Outline of Production for a

biological product recommended for use

in dogs. Serials which are not found to

be satisfactory when tested pursuant to
Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations 14359

the procedures in this section may not

be released for shipment.

(a) The dog safety test provided in this

paragraph shall be used when the

Master Seed Virus is tested for safety.

(1) The test animals shall be

determined to be susceptible to the

virus under test by a method acceptable

to the Animal and Plant Health

Inspection Service.

(2) Each of at least 10 susceptible dogs

shall be administered a sample of the

Master Seed Virus equivalent to the

amount of virus to be used in one dog

dose of the vaccine, by the method

recommended on the label, and the dog

shall be observed each day for 14 days.

(3) If unfavorable reactions

attributable to the virus occur in any of

the dogs during the observation period,

the Master Seed Virus is unsatisfactory.

If unfavorable reactions occur which are

not attributable to the Master Seed

Virus, the test shall be declared

inconclusive and may be repeated:

Provided: That, if the test is not

repeated, the Master Seed Virus shall be

considered unsatisfactory.

(b) The dog safety test provided in

this paragraph shall be used when a

serial of vaccine is tested for safety

before release.

(1) Each of two healthy dogs shall be

administered 10 dog doses by the

method recommended on the label and

the dogs shall be observed each day for

14 days.

(2) If unfavorable reactions

attributable to the biological product

occur during the observation period, the

serial is unsatisfactory. If unfavorable

reactions occur which are not

attributable to the biological product,

the test shall be declared inconclusive

and may be repeated: Provided, That, if

the test is not repeated, the serial shall

be considered unsatisfactory.

3. Section 113.201 is revised to read

as follows:

§ 113.201 Canine Distemper Vaccine,

Killed Virus.

Canine Distemper Vaccine, Killed

Virus, shall be prepared from virusbearing cell culture fluids. Only Master

Seed Virus which has been established

as pure, safe, and immunogenic shall be

used for vaccine production. All serials

of vaccine shall be prepared from the

first through the fifth passage from the

Master Seed Virus.

(a) The Master Seed Virus shall meet

the applicable general requirements

prescribed in § 113.200.

(b) The immunogenicity of vaccine

prepared from the Master Seed Virus in

accordance with the Outline of

Production shall be established. Vaccine

used for this test shall be at the highest

passage from the Master Seed and

prepared at the minimum

preinactivation titer specified in the

Outline of Production.

(1) Twenty-five canine distemper

susceptible dogs (20 vaccinates and 5

controls) shall be used as test animals.

Blood samples drawn from each dog

shall be individually tested for

neutralizing antibody against canine

distemper to determine susceptibility. A

constant virus-varying serum

neutralization test in cell culture using

50 to 300 TCID50 of virus shall be used.

Dogs shall be considered susceptible if

there is no neutralization at a 1:2 final

serum dilution.

(i) The 20 dogs used as vaccinates

shall be injected with one dose of

vaccine by the method recommended on

the label. If a second dose is

recommended, the second dose shall be

administered at the time specified on

the label.

(ii) At least 14 days after the last

inoculation, the vaccinates and controls

shall each be challenged intracerebrally

with canine distemper virus furnished

or approved by the Animal and Plant

Health Inspection Service and observed

each day for 21 days.

(iii) If at least four of the five controls

do not die and the survivor, if any, does

not show clinical signs of canine

distemper, the test is inconclusive and

may be repeated.

(iv) If at least 19 of the 20 vaccinated

do not survive without showing clinical

signs of canine distemper during the

observation period, the Master Seed

Virus is unsatisfactory.

(c) Test requirements for release. Each

serial shall meet the applicable general

requirements prescribed in § 113.200

and the special requirements for safety

and potency provided in this section.

(1) Safety test. The vaccinates used in

the potency test in paragraph (c)(2) of

this section shall be observed each day

during the postvaccination observation

period. If unfavorable reactions occur

which are attributable to the vaccine,

the serial is unsatisfactory. If

unfavorable reactions occur which are

not attributable to the vaccine, the test

is inconclusive and may be repeated:

Provided, That, if the test is not

repeated, the serial is unsatisfactory.

(2) Potency test—serum neutralization

test. Bulk or final container samples of

completed product shall be tested for

potency using five susceptible dogs

(four vaccinates and one control) as the

test animals. Blood samples drawn from

each dog shall be individually tested for

neutralizing antibody against canine

distemper virus to determine

susceptibility.

(i) A constant virus-varying serum

neutralization test in tissue culture

using 50 to 300 TCID50 of virus shall be

used. Dogs shall be considered

susceptible if there is no neutralization

at a 1:2 final serum dilution.

(ii) Vaccination. Each of the four

vaccinates shall be injected as

recommended on the label. If two doses

are recommended, the second dose shall

be administered at the time specified on

the label. The dogs shall be observed

each day for at least 14 days after the

last inoculation.

(iii) Serology. At the end of the post

vaccination observation period, a

second blood sample shall be obtained

from each of the five dogs and the

serums shall be individually tested for

neutralizing antibody against canine

distemper virus in the same manner

used to determine susceptibility.

(iv) Interpretation of the serum

neutralization test. If the control has not

remained seronegative at 1:2, the test is

inconclusive and may be repeated. If at

least three of the four vaccinates in a

valid test have not developed titers

based upon a final serum dilution of at

least 1:50 and the remaining vaccinate

has not developed a titer of at least 1:25,

the serial is unsatisfactory except as

provided in paragraphs (c)(2) (v) and

(vi) of this section.

(v) Virus challenge test. If the results

of a valid serum neutralization test are

unsatisfactory, the vaccinates and the

control may be challenged

intracerebrally with a virulent canine

distemper virus furnished or approved

by the Animal and Plant Health

Inspection Service and each animal

observed each day for an additional 21

days.

(vi) Interpretation of the virus

challenge test. For a serial to be

satisfactory, all vaccinates must remain

free from clinical signs of canine

distemper while the control must die of

canine distemper. If the control does not

die of canine distemper, the test is

inconclusive and may be repeated

except, that if any of the vaccinates

show signs or dies of canine distemper,

the serial is unsatisfactory.

4. Section 113.202 is revised to read

as follows:

§ 113.202 Canine Hepatitis and Canine

Adenovirus Type 2 Vaccine, Killed Virus.

Canine Hepatitis and Canine

Adenovirus Type 2 Vaccine, Killed

Virus, shall be prepared from virusbearing cell culture fluids. Only Master

Seed Virus which has been established

as pure, safe, and immunogenic shall be

used for vaccine production. All serials

of vaccine shall be prepared from the
14360 Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations

first through the fifth passage from the

Master Seed Virus.

(a) The Master Seed Virus shall meet

the applicable requirements prescribed

in § 113.200.

(b) Each lot of Master Seed Virus used

for vaccine production shall be tested

for immunogenicity by one or both of

the following methods. Vaccine used for

these tests shall be at the highest

passage from the Master Seed and

prepared at the minimum

preinactivation titer specified in the

Outline of Production.

(1) Immunogenicity for canine

hepatitis. Twenty-five canine hepatitis

susceptible dogs shall be used as test

animals (20 vaccinates and 5 controls).

Blood samples shall be drawn from

these animals and individual serum

samples tested. The dogs shall be

considered susceptible if the results are

negative at a 1:2 final serum dilution in

a varying serum-constant virus

neutralization test using 50 to 300

TCID50 of canine adenovirus.

(i) The 20 dogs to be used as

vaccinates shall be injected with one

dose of vaccine and the remaining five

dogs held as controls. If a second dose

is recommended, the second dose shall

be administered at the time specified on

the label.

(ii) Not less than 14 days after the last

inoculation, each vaccinate and control

shall be challenged intravenously with

virulent infectious canine hepatitis

virus furnished or approved by the

Animal and Plant Health Inspection

Service and observed each day for 14

days.

(iii) If at least four of the five controls

do not show severe clinical signs of

infectious canine hepatitis, the test is

inconclusive and may be repeated.

(iv) If at least 19 of the 20 vaccinates

do not survive without showing clinical

signs of infectious canine hepatitis

during the observation period, the

Master Seed Virus is unsatisfactory.

(2) Immunogenicity for canine

adenovirus type 2. Thirty canine

adenovirus type 2 susceptible dogs shall

be used as test animals (20 vaccinates

and 10 controls). Blood samples shall be

drawn from these animals and

individual serum samples tested. The

dogs shall be considered susceptible if

the results are negative at a 1:2 final

serum dilution in a varying serumconstant virus neutralization test using

50 to 300 TCID50 of canine adenovirus.

(i) The 20 dogs to be used as

vaccinates shall be injected with one

dose of vaccine and the remaining 10

dogs held as controls. If a second dose

is recommended, the second dose shall

be administered at the time specified on

the label.

(ii) Not less than 14 days after the last

inoculation, the vaccinates and the

controls shall be challenged by exposure

to a nebulized aerosol of virulent canine

adenovirus type 2 furnished or

approved by the Animal and Plant

Health Inspection Service and observed

each day for 14 days postchallenge. The

rectal temperature of each animal shall

be taken and the presence of respiratory

or other clinical signs of canine

adenovirus type 2 noted and recorded

each day.

(iii) If at least 6 of 10 controls do not

show clinical signs of canine adenovirus

type 2 infection other than fever, the test

is inconclusive and may be repeated.

(iv) If a significant difference in

clinical signs in a valid test cannot be

demonstrated between vaccinates and

controls using a scoring system

approved by the Animal and Plant

Health Inspection Service, the Master

Seed Virus is unsatisfactory.

(c) Test requirements for release. Each

serial shall meet the applicable general

requirements prescribed in § 113.200,

the special requirements for safety

provided in this section, and the

applicable potency tests provided in

this section.

(1) Safety test. The vaccinates used in

the potency test in paragraph (c)(2) and/

or (c)(3) of this section shall be observed

each day during the postvaccination

observation period. If unfavorable

reactions occur which are attributable to

the vaccine, the serial is unsatisfactory.

If unfavorable reactions occur which are

not attributable to the vaccine, the test

is inconclusive and may be repeated:

Provided, That, if not repeated, the

serial is unsatisfactory.

(2) Potency test for canine hepatitis—

serum neutralization test. Bulk or final

container samples of completed product

shall be tested for potency using at least

five susceptible dogs (four vaccinates

and one control) as the test animals.

Blood samples drawn from each dog

shall be individually tested for

neutralizing antibody against canine

adenovirus to determine susceptibility.

(i) A constant virus-varying serum

neutralization test in tissue culture

using 50 to 300 TCID50 of virus shall be

used. Dogs shall be considered

susceptible if there is no neutralization

at a 1:2 final serum dilution.

(ii) Vaccination. Each of the

vaccinates shall be injected as

recommended on the label. If two doses

are recommended, the second dose shall

be administered at the time specified on

the label. The dogs shall be observed

each day for at least 14 days after the

last inoculation.

(iii) Serology. At the end of the

postvaccination observation period, a

second blood sample shall be obtained

from each of the dogs and the serums

shall be individually tested for

neutralizing antibody against canine

adenovirus in the same manner used to

determine susceptibility.

(iv) Interpretation of the serum

neutralization test. If the control(s) has

not remained seronegative at 1:2, the

test is inconclusive and may be

repeated. If at least 75 percent of the

vaccinates in a valid test have not

developed titers based upon final serum

dilution of at least 1:10 and the

remaining vaccinate(s) has not

developed a titer of at least 1:2, the

serial is unsatisfactory except as

provided in paragraphs (c)(2) (v) and

(vi) of this section.

(v) Virus challenge test. If the results

of a valid serum neutralization test are

unsatisfactory, the vaccinates and the

control(s) may be challenged

intravenously with a virulent canine

hepatitis virus furnished or approved by

the Animal and Plant Health Inspection

Service and each animal observed each

day for an additional 14 days.

(vi) Interpretation of the virus

challenge test. For a serial to be

satisfactory, all vaccinates must remain

free of clinical signs of canine hepatitis

while the control(s) must show severe

clinical signs of canine hepatitis. If the

control(s) does not show severe clinical

signs of canine hepatitis, the test is

inconclusive and may be repeated:

Provided, That, if any of the vaccinates

show signs or die of canine hepatitis,

the serial is unsatisfactory.

(3) Potency test for canine adenovirus

type 2. Bulk or final container samples

of completed product shall be tested for

potency using eight susceptible dogs

(five vaccinates and three controls) as

the test animals. Blood samples drawn

from each dog shall be individually

tested for neutralizing antibody against

canine adenovirus to determine

susceptibility.

(i) A constant virus-varying serum

neutralization test in tissue culture

using 50 to 300 TCID50 of virus shall be

used. Dogs shall be considered

susceptible if there is no neutralization

at a 1:2 final serum dilution.

(ii) Vaccination. Each of the five

vaccinates shall be injected as

recommended on the label. If two doses

are recommended, the second dose shall

be administered at the time specified on

the label. The dogs shall be observed

each day for at least 14 days after the

last inoculation.

(iii) Not less than 14 days after the last

inoculation, the vaccinates and the

controls shall be challenged by exposure

to a nebulized aerosol of virulent canine

adenovirus type 2 furnished or
Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations 14361

approved by the Animal and Plant

Health Inspection Service and observed

each day for 14 days postchallenge. The

rectal temperature of each animal shall

be taken and the presence of respiratory

or other clinical signs of canine

adenovirus type 2 noted and recorded

each day.

(iv) If at least two of three controls do

not show clinical signs of canine

adenovirus type 2 other than fever, the

test is inconclusive and may be

repeated.

(v) If a significant difference in

clinical signs cannot be demonstrated

between vaccinates and controls using a

scoring system approved by the Animal

and Plant Health Inspection Service and

prescribed in the Outline of Production,

the serial is unsatisfactory.

5. Section 113.204 is amended by

revising paragraphs (b)(2) and (b)(3) to

read as follows:

§ 113.204 Mink Enteritis Vaccine, Killed

Virus.

* * * * *

(b) * * *

(2) Challenge. At least 2 weeks after

the last inoculation, the five vaccinates

and the five controls shall be challenged

with virulent mink enteritis virus and

observed each day for 12 days. Fecal

material shall be collected on one day

between days 4–8 (inclusive)

postchallenge from each test animal that

remains free of enteric signs and tested

for the presence of mink enteritis virus

by cell culture with fluorescent

antibody examination.

(3) Interpretation. A serial is

satisfactory if at least 80 percent of the

vaccinates remain free of enteric signs

and do not shed virus in the feces, while

at least 80 percent of the controls

develop clinical signs of mink enteritis

or shed virus in the feces. If at least 80

percent of the vaccinates remain free of

enteric signs and do not shed virus in

the feces, while less than 80 percent of

the controls develop clinical signs of

mink enteritis or shed virus in the feces,

the test is considered inconclusive and

may be repeated: Provided, That, if at

least 80 percent of the vaccinates do not

remain well and free of detectable virus

in the feces, the serial is unsatisfactory.

6. Section 113.305 is revised to read

as follows:

§ 113.305 Canine Hepatitis and Canine

Adenovirus Type 2 Vaccine.

Canine Hepatitis Vaccine and Canine

Adenovirus Type 2 Vaccine shall be

prepared from virus-bearing cell culture

fluids. Only Master Seed Virus which

has been established as pure, safe, and

immunogenic shall be used in preparing

the production seed virus for vaccine

production. All serials shall be prepared

from the first through the fifth passage

from the Master Seed Virus.

(a) The Master Seed Virus shall meet

the applicable requirements prescribed

in § 113.300 except that the dog safety

test prescribed in § 113.40(a) shall be

conducted by the intravenous route.

(b) Each lot of Master Seed Virus used

for vaccine production shall be tested

for immunogenicity by one or both of

the following methods:

(1) Immunogenicity for canine

hepatitis. Twenty-five canine hepatitis

susceptible dogs shall be used as test

animals (20 vaccinates and 5 controls).

Blood samples shall be drawn from

these animals and individual serum

samples tested. The dogs shall be

considered susceptible if the results are

negative at a 1:2 final serum dilution in

a varying serum-constant virus

neutralization test using 50 to 300

TCID50 of canine adenovirus.

(i) A geometric mean titer of the dried

vaccine produced from the highest

passage of the Master Seed Virus shall

be established before the

immunogenicity test is conducted. The

20 dogs to be used as vaccinates shall

be injected with a predetermined

quantity of vaccine virus and the

remaining five dogs held as uninjected

controls. To confirm the dosage

calculations, five replicate virus

titrations shall be conducted on a

sample of the vaccine virus dilution

used.

(ii) Not less than 14 days

postinjection, the vaccinates and the

controls shall each be challenged

intravenously with virulent infectious

canine hepatitis virus furnished or

approved by the Animal and Plant

Health Inspection Service and observed

each day for 14 days.

(A) If at least four of the five controls

do not show severe clinical signs of

canine hepatitis, the test is inconclusive

and may be repeated.

(B) If at least 19 of the 20 vaccinates

do not survive without showing clinical

signs of infectious canine hepatitis

during the observation period, the

Master Seed Virus is unsatisfactory.

(iii) The Master Seed Virus shall be

retested for immunogenicity for canine

hepatitis in 3 years unless use of the lot

previously tested is discontinued. Ten

susceptible dogs (8 vaccinates and 2

controls) shall be used in the retest.

Susceptibility shall be determined in

the manner provided in paragraph (b)(1)

of this section.

(A) Each vaccinate shall be injected

with a predetermined quantity of

vaccine virus as provided in paragraph

(b)(1)(i) of this section.

(B) At least 14 days postvaccination,

a second serum sample shall be drawn

from each dog and tested for

neutralizing antibody to canine

adenovirus in the same manner used to

determine susceptibility.

(C) If the two controls have not

remained seronegative at 1:2, the test is

inconclusive and may be repeated.

(D) if at least six of the eight

vaccinates in a valid test do not develop

titers of at least 1:10 based upon final

serum dilution, the Master Seed Virus is

unsatisfactory except as provided in

paragraph (b)(1)(iii)(E) of this section.

(E) if the results of a valid serum

neutralization test are unsatisfactory,

the vaccinates and the controls may be

challenged as provided in paragraph

(b)(1)(ii) of this section. A Master Seed

is satisfactory if all vaccinates remain

free of clinical signs of canine hepatitis,

while both controls develop severe

clinical signs of canine heptatis. If both

controls do not show severe clinical

signs of canine hepatitis, the test is

inconclusive and may be repeated:

Provided, That, if any of the vaccinates

show such signs, the Master Seed Virus

is unsatisfactory.

(2) Immunogenicity for canine

adenovirus Type 2. Thirty canine

adenovirus type 2 susceptible dogs shall

be used as test animals (20 vaccinates

and 10 controls). Blood samples shall be

drawn from these animals and

individual serum samples tested. The

dogs shall be considered susceptible if

the results are negative at a 1:2 final

serum dilution in a varying serumconstant virus neutralization test using

50 to 300 TCID50 of canine adenovirus.

(i) A geometric mean titer of the dried

vaccine produced from the highest

passage of the Master Seed Virus shall

be established before the

immunogenicity test is conducted. The

20 dogs to be used as vaccinates shall

be injected with a predetermined

quantity of vaccine virus and the

remaining 10 dogs held as uninjected

controls. To confirm the dosage

calculations, five replicate virus

titrations shall be conducted on a

sample of the vaccine virus dilution

used.

(ii) Not less than 14 days

postinjection, the vaccinates and the

controls shall be challenged by exposure

to a nebulized aerosol of virulent canine

adenovirus type 2 furnished or

approved by the Animal and Plant

Health Inspection Service and observed

each day for 14 days postchallenge. The

rectal temperature of each animal shall

be taken and the presence of respiratory

or other clinical signs of canine

adenovirus type 2 noted and recorded

each day.
14362 Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations

(A) If at least 6 of 10 controls do not

show clinical signs of canine adenovirus

type 2 infection other than fever, the test

is inconclusive and may be repeated.

(B) if a significant difference in

clinical signs in a valid test cannot be

demonstrated between vaccinates and

controls using a scoring system

approved by the Animal and Plant

Health Inspection Service, the Master

Seed Virus is unsatisfactory.

(iii) the Master Seed Virus shall be

retested for immunogenicity in 3 years

unless use of the lot previously tested is

discontinued. Either 10 vaccinates and

6 controls or 5 vaccinates and 3 controls

shall be used in the retest.

(A) If less than 4 of 6 or 2 of 3 of the

controls show clinical signs of canine

adenovirus type 2 other than fever, the

test is inconclusive and may be

repeated.

(B) a significant difference in clinical

signs shall be demonstrated between

vaccinates and controls in a valid test as

prescribed in paragraph (b)(2)(ii)(B) of

this section.

(iv) an Outline of Production change

shall be made before authorization for

use of a new lot of Master Seed Virus

shall be granted by the Animal and

Plant Health Inspection Service.

(c) Test requirements for release. Each

serial and subserial shall meet the

requirements prescribed in § 113.300

and in this paragraph. Final container

samples of completed product shall be

tested. Any serial or subserial found

unsatisfactory by a prescribed test shall

not be released.

(1) Virus titer requirements. Final

container samples of completed product

shall be tested for virus titer using the

titration method used in paragraph

(b)(1)(i) and/or (b)(2)(i) of this section.

To be eligible for release, each serial and

each subserial shall have a virus titer

sufficiently greater than the titer of

vaccine virus used in the

immunogenicity test(s) prescribed in

paragraph (b) of this section to assure

that when tested at any time within the

expiration period, each serial and

subserial shall have a virus titer of 100.7

greater than that used in such

immunogenicity test(s) but not less than

10.2.5 TCID50 dose. If both

immunogenicity tests in paragraph (b) of

this section are conducted and a

different amount of virus is used in each

test, the virus titer requirements shall be

based on the higher of the two amounts.

7. Section 113.306 is revised to read

as follows:

§ 113.306 Canine Distemper Vaccine.

Canine Distemper Vaccine shall be

prepared from virus-bearing cell culture

fluids or embryonated chicken eggs.

Only Master Seed Virus which has been

established as pure, safe, and

immunogenic shall be used for

preparing the production seed virus for

vaccine production. All serials of

vaccine shall be prepared from the first

through the fifth passage from the

Master Seed Virus.

(a) Master Seed Virus. The Master

Seed Virus shall meet the applicable

requirements prescribed in § 113.300

and the requirements prescribed in this

section.

(1) To detect ferret virulent canine

distemper virus, each of five canine

distemper susceptible ferrets shall be

injected with a sample of the Master

Seed Virus equivalent to the amount of

virus to be used in one dog dose and

observed each day for 21 days. If

undesirable reactions are observed

during the observation period, the lot of

Master Seed is unsatisfactory.

(2) Master Seed Virus propagated in

tissues or cells of avian origin shall be

tested for pathogens by the chicken

embryo test prescribed in § 113.37. If

found unsatisfactory, the Master Seed

Virus shall not be used.

(b) Each lot of Master Seed Virus used

for vaccine production shall be tested

for immunogenicity. The selected virus

dose from the lot of Master Seed Virus

shall be established as follows:

(1) Twenty-five canine distemper

susceptible dogs shall be used as test

animals (20 vaccinates and 5 controls).

Blood samples shall be drawn from

these animals and individual serum

samples tested. The dogs shall be

considered susceptible if the results are

negative at a 1:2 final serum dilution in

a varying serum-constant virus

neutralization test using 50 to 300

TCID50 of canine distemper virus.

(2) A geometric mean titer of the dried

vaccine produced from the highest

passage of the Master Seed Virus shall

be established before the

immunogenicity test is conducted. The

20 dogs used as vaccinates shall be

injected with a predetermined quantity

of vaccine virus and the remaining five

dogs held as uninjected controls. To

confirm the dosage calculations, five

replicate virus titrations shall be

conducted on a sample of the vaccine

virus dilution used.

(3) At least 14 days post-injection, the

vaccinates and the controls shall each

be challenged intracerebrally with

virulent canine distemper virus

furnished or approved by the Animal

and Plant Health Inspection Service and

observed each day for 21 days.

(i) If at least four of the five controls

do not die and the survivor, if any does

not show clinical signs of canine

distemper the test is inconclusive and

may be repeated.

(ii) If at least 19 of the 20 vaccinates

do not survive without showing clinical

signs of canine distemper during the

observation period, the Master Seed

Virus is unsatisfactory.

(4) The Master Seed Virus shall be

retested for immunogenicity in 3 years

unless use of the lot previously tested is

discontinued. Ten susceptible dogs (8

vaccinates and 2 controls) shall be used

in the retest. Susceptibility shall be

determined in the manner provided in

paragraph (b)(1) of this section.

(i) Each vaccinate shall be injected

with a predetermined quantity of

vaccine virus as provided in paragraph

(b)(2) of this section.

(ii) At least 14 days postvaccination,

a second serum sample shall be drawn

from each dog and tested for

neutralizing antibody to canine

distemper virus in the same manner

used to determine susceptibility.

(iii) If the two controls have not

remained seronegative at 1:2, the test is

inconclusive and may be repeated.

(iv) If at least 6 of the 8 vaccinates in

a valid test do not develop titers of at

least 1:50 based upon final serum

dilution, the Master Seed Virus is

unsatisfactory, except as provided in

paragraph (b)(4)(v) of this section.

(v) If the results of a valid serum

neutralization test are unsatisfactory,

the vaccinates and the controls may be

challenged as provided in paragraph

(b)(3) of this section. A Master Seed is

satisfactory if all vaccinates remain free

of clinical signs of canine distemper,

while the two controls die with clinical

signs of canine distemper. If the two

controls do not die with clinical signs

of canine distemper, the test is

inconclusive and may be repeated:

Provided, That, if any of the vaccinates

show such signs, the Master Seed Virus

is unsatisfactory.

(5) An Outline of Production change

shall be made before authorization for

use of a new lot of Master Seed Virus

shall be granted by the Animal and

Plant Health Inspection Service.

(c) Test requirements for release.

Except for § 113.300(a)(3)(ii), each serial

and subserial shall meet the

requirements prescribed in § 113.300

and in this paragraph. Final container

samples of completed product shall be

tested. Any serial or subserial found

unsatisfactory by a prescribed test shall

not be released.

(1) The test for pathogens prescribed

in § 113.37 shall be conducted on each

serial or one subserial of avian origin

vaccine.

(2) Virus titer requirements. Final

container samples of completed product
Federal Register / Vol. 60, No. 52 / Friday, March 17, 1995 / Rules and Regulations 14363

shall be tested for virus titer using the

titration method used in paragraph

(b)(2) of this section. To be eligible for

release, each serial and subserial shall

have a virus titer sufficiently greater

than the titer of vaccine virus used in

the immunogenicity test prescribed in

paragraph (b) of this section to assure

that when tested at any time within the

expiration period, each serial and

subserial shall have a virus titer of 100.7

greater than that used in such

immunogenicity test but not less than

102.5 TCID50 per dose.

§ 113.307 [Removed]

8. Section 113.307 is removed.

Done in Washington, DC, this 13th day of

March 1995.

Terry L. Medley,

Acting Administrator, Animal and Plant

Health Inspection Service.

[FR Doc. 95–6647 Filed 3–16–95; 8:45 am]

BILLING CODE 3410–34–M

DEPARTMENT OF TRANSPORTATION

Federal Aviation Administration

14 CFR Part 71

[Airspace Docket No. 93–ACE–02]

Amendment to Class E Airspace;

Harvard, NE

AGENCY: Federal Aviation

Administration [FAA], DOT.

ACTION: Final rule.

SUMMARY: This amendment modifies the

Class E airspace area at Harvard, NE to

accommodate a planned Very High

Frequency Omnidirectional Range/

Distance Measuring Equipment (VOR/

DME) Standard Instrument Approach

Procedure (SIAP) at the Harvard State

Airport. This action will provide for

additional controlled airspace necessary

for the planned VOR/DME SIAP. It will

also change the airport status from

Visual Flight Rules (VFR) to Instrument

Flight Rules (IFR).

EFFECTIVE DATE: 0901 UTC May 25,

1995.

FOR FURTHER INFORMATION CONTACT:

Kathy Randolph, Air Traffic Operations

Branch, ACE–530c, Federal Aviation

Administration, 6021 E. 12th St., Kansas

City, MO, 64106; telephone (816) 426–

3408.

SUPPLEMENTARY INFORMATION:

History

On January 7, 1994, the FAA

proposed to amend part 71 of the

Federal Aviation Regulations (14 CFR

part 71) by modifying the Class E

airspace area at Harvard, NE (59 FR

3032). The proposed action would

provide additional controlled airspace

to accommodate a VOR/DME SIAP to

Runway 35 at the Harvard State Airport.

Interested parties were invited to

participate in this rulemaking

proceeding by submitting written

comments on the proposal to the FAA.

No comments objecting to the proposal

were received. Class E airspace areas

extending from 700 feet or more above

the surface of the earth are published in

paragraphs 6005 of FAA order 7400.9B,

dated July 8, 1994, and effective

September 16, 1994, which is

incorporated by reference in 14 CFR

71.1. The Class E airspace designation

listed in this document will be

published subsequently in the Order.

The Rule

This amendment to part 71 of the

Federal Aviation Regulations (14 CFR

part 71) amends the Class E airspace

area at Harvard, NE, by providing

additional controlled airspace for

aircraft executing the VOR/DME runway

35 SIAP to the Harvard State Airport.

The FAA has determined that this

regulation only involves an established

body of technical regulations for which

frequent and routine amendments are

necessary to keep them operationally

current. Therefore, this regulation—(1)

is not a ‘‘significant regulatory action’’

under Executive Order 12866; (2) is not

a ‘‘significant rule’’ under DOT

Regulator Policies and Procedures (44

FR 11034; February 26, 1979); and (3)

does not warrant preparation of a

Regulatory Evaluation as the anticipated

impact is so minimal. Since this is a

routine matter that will only affect air

traffic procedures and air navigation, it

is certified that this rule will not have

a significant economic impact on a

substantial number of small entities

under the criteria of the Regulatory

Flexibility Act.

List of Subjects in 14 CFR Part 71

Aviation, Incorporation by reference,

Navigation (air).

Adoption of the Amendment

In consideration of the foregoing, the

Federal Aviation Administration

amends 14 CFR part 71 as follows:

PART 71—[AMENDED]

1. The authority citation for part 71

continues to read as follows:

Authority: 49 U.S.C. app. 1348(a), 1354(a),

1510; E.O. 10854, 24 FR 9565, 3 CFR, 1959–

1963 Comp., p. 389; 49 U.S.C. 106(g); 14 CFR

11.69.

§ 71.1 [Amended]

2. The incorporation by reference in

14 CFR 71.1 of Federal Aviation

Administration Order 7400.9B, Airspace

Designations and Reporting Points,

dated July 18, 1994 and effective

September 16, 1994, is amended as

follows:

Paragraph 6005 Class E airspace areas

extending from 700 feet or more above the

surface of the earth.

* * * * *

ACE NE E5 Harvard, NE [Revised]

Harvard State Airport, NE

(Lat. 40°39′15′′ N, long. 98°04′31′′ W)

That airspace extending upward from 700

feet above the surface within 6.4-mile radius

of the Harvard State Airport and within 2

miles each side of the 180° bearing of the

Harvard State Airport extending from the 6.4-

mile radius to 10 miles south of the airport.

* * * * *

Issued in Kansas City, MO on February 21,

1995.

Clarence E. Newbern,

Manager, Air Traffic Division, Central Region.

[FR Doc. 95–6685 Filed 3–16–95; 8:45 am]

BILLING CODE 4910–13–M

14 CFR Part 97

[Docket No. 28135; Amdt. No. 1654]

Standard Instrument Approach

Procedures; Miscellaneous

Amendments

AGENCY: Federal Aviation

Administration (FAA), DOT.

ACTION: Final rule.

SUMMARY: This amendment establishes,

amends, suspends, or revokes Standard

Instrument Approach Procedures

(SIAPs) for operations at certain

airports. These regulatory actions are

needed because of the adoption of new

or revised criteria, or because of changes

occurring in the National Airspace

System, such as the commissioning of

new navigational facilities, addition of

new obstacles, or changes in air traffic

requirements. These changes are

designed to provide safe and efficient

use of the navigable airspace and to

promote safe flight operations under

instrument flight rules at the affected

airports.

DATES: An effective date for each SIAP

is specified in the amendatory

provisions.

Incorporation by reference-approved

by the Director of the Federal Register

on December 31, 1980, and reapproved

as of January 1, 1982.

ADDRESSES: Availability of matters

incorporated by reference in the

amendment is as follows: