[Federal Register Volume 60, Number 52 (Friday, March 17, 1995)]
[Rules and Regulations]
[Pages 14357-14363]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 95-6647]
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DEPARTMENT OF AGRICULTURE
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.
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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 Sec. 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 Sec. 113.204(b)(2) to specify that feces are [[Page 14358]] 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 Sec. 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 Sec. 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 (Secs. 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 Sec. 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 Sec. 113.306. An
intracerebral challenge has been used successfully for many years with
the live virus vaccine. It was the proposed amendments to Sec. 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 Sec. 113.306. Since the commenters focussed only on
Sec. 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 Sec. 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:
Sec. 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
[[Page 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:
Sec. 113.201 Canine Distemper Vaccine, Killed Virus.
Canine Distemper Vaccine, Killed Virus, 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 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 Sec. 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 Sec. 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:
Sec. 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 virus-bearing 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 [[Page 14360]] first through the
fifth passage from the Master Seed Virus.
(a) The Master Seed Virus shall meet the applicable requirements
prescribed in Sec. 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 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 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 Sec. 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
[[Page 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:
Sec. 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:
Sec. 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 Sec. 113.300 except that the dog safety test prescribed
in Sec. 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 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 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. [[Page 14362]]
(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 Sec. 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:
Sec. 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 Sec. 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 Sec. 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
Sec. 113.300(a)(3)(ii), each serial and subserial shall meet the
requirements prescribed in Sec. 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 Sec. 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 [[Page 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.
Sec. 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
