[Federal Register Volume 59, Number 195 (Tuesday, October 11, 1994)]
[Unknown Section]
[Page 0]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 94-25056]
[[Page Unknown]]
[Federal Register: October 11, 1994]
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DEPARTMENT OF AGRICULTURE
Animal and Plant Health Inspection Service
9 CFR Part 113
[Docket No. 93-039-1]
Viruses, Serums, Toxins and Analogous Products; Standard
Requirement for Escherichia Coli Bacterins
AGENCY: Animal and Plant Health Inspection Service, USDA.
ACTION: Proposed rule.
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SUMMARY: We are proposing to amend the regulations by adding a Standard
Requirement for Escherichia coli bacterins. This amendment would
provide a uniform procedure to demonstrate the immunogenicity of
reference bacterins made from E. coli master seed bacteria. This
amendment would also provide uniform procedures to requalify a
reference bacterin for which the dating period has expired. In
addition, the amendment would specify the potency test requirements for
the release of serials of E. coli bacterins for immunogenicity and
potency.
The purpose of this regulation is to standardize the requirements
for the production and testing of E. coli bacterins for immunogenicity
and potency.
DATES: Consideration will be given only to comments received on or
before December 12, 1994.
ADDRESSES: Please send an original and three copies of your comments to
Chief, Regulatory Analysis and Development, PPD, APHIS, USDA, room 804,
Federal Building, 6505 Belcrest Road, Hyattsville, MD 20782. Please
state that your comments refer to Docket No. 93-039-01. Comments
received may be inspected at USDA, room 1141, South Building, 14th
Street and Independence Avenue SW., Washington, DC, between 8 a.m. and
4:30 p.m., Monday through Friday, except holidays. Persons wishing to
inspect comments are requested to call ahead (202) 690-2817 to
facilitate entry into the comment reading room.
FOR FURTHER INFORMATION CONTACT:
Dr. David A. Espeseth, Deputy Director, Veterinary Biologics, BBEP,
APHIS, USDA, room 838, Federal Building, 6505 Belcrest Road,
Hyattsville, MD 20782, (301) 436-8245.
SUPPLEMENTARY INFORMATION:
Background
Standard Requirements are prescribed in 9 CFR part 113 for the
preparation and testing of veterinary biological products. A Standard
Requirement consists of test methods, procedures, and criteria which
define the standards of purity, safety, potency, and efficacy for a
given type of veterinary biological product. Where a Standard
Requirement for a product does not exist in the regulations, production
procedures and specifications for purity, safety, and potency of a
product are provided in an Outline of Production filed with the Animal
and Plant Health Inspection Service. Once uniform standards for a type
of product are established, they are codified in the regulations.
Because there is no Standard Requirement in 9 CFR part 113 for
Escherichia coli (E. coli) bacterins, each manufacturer of these
products has devised its own procedures, which are usually a part of a
filed Outline of Production, to meet the requirements of the Virus-
Serum-Toxin Act that all veterinary biological products be pure, safe,
potent, and efficacious. Therefore, even though all of the procedures
and methods must be satisfactory to the Animal and Plant Health
Inspection Service, there are a number of different procedures which
are used by firms producing biological products containing E. coli as
one of the components. At this time, we are prepared to propose adding
a new Sec. 113.124 to the standards which would contain uniform test
methods, procedures, and criteria to be used by licensed manufacturers
of E. coli bacterins to provide assurance that these bacterins are
immunogenic and potent.
Immunogencity
The effectiveness of a veterinary biologic to ameliorate disease in
animals is of paramount importance to the consumers of veterinary
biological products. The test for immunogenicity ascertains the
efficacy of a veterinary biologic. Based on experience with
immunogenicity tests, the Animal and Plan Health Inspection Service is
proposing uniform requirements for establishing the immunogenicity of a
Master Reference made from E. coli master seed. This would be
accomplished by specifying in proposed Sec. 113.124(a): (1) The minimum
number of pregnant dams to vaccinate and the number of nonvaccinated
controls; (2) procedures for challenge of neonates with virulent E.
coli; and (3) the criteria for satisfactory protection against
challenge and thereby demonstration of immunogenicity of the Master
Reference. The immunogenicity test of master seed bacteria described in
proposed Sec. 113.124(a) would be required of all new product license
applications prior to licensure. The test would establish the
immunogenicity of the Master Reference used in the relative potency
test required for release of serials of biological product.
As do all biologics, reference bacterins may lose potency and
immunogenicity with the passage of time. The loss of potency and
immunogenicity is not constant for all biologics because of differences
between master seeds, production methods, adjuvants, diluents,
conditions of storage and other variables used in the manufacture of
biologics. Therefore the proposed amendment in Sec. 113.124(b)
describes the testing that must be conducted on an E. coli Master
Reference to determine if it is still sufficiently potent and
immunogenic after its date of expiration to permit extending the date
of expiration.
Potency
Different potency tests are currently used for E. coli bacterns.
Potency tests used to compare serials of product to a reference
bacterin are: mouse tests, guinea pig tests, and enzyme linked
immunosorbent assays (ELISA). As proposed, new Sec. 113.124(c) would
require the use of a standard in vitro method for testing serials of E.
coli bacterins. The proposed standard method is a parallel line
immunoassay that compares, for both the unknown and the reference, the
linear regression of the optical density (OD) versus the logarithm of
antigen concentration determined by ELISA. The Animal and Plant Health
Inspection Service has proposed use of this standard in vitro potency
test for E. coli bacterins
because its use provides three advantages: (1) It ensures the use of a
valid test method and makes potency testing more uniform; (2) it
ensures that serials of product are potent before release; and (3) it
reduces significantly the number of laboratory animals required for
testing E. coli bacterins.
If adopted, this standard test would require that manufacturing
firms have personnel proficient in performing the ELISA procedure and
the equipment necessary to perform the tests.
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 purposes of
Executive Order 12866, and, therefore, has not been reviewed by the
Office of Management and Budget.
This proposed rule, if adopted, would aid firms manufacturing E.
coli bacterins. The proposal contains Standard Requirements for
immunogenicity testing which would provide uniformity among firms
instead of each firm having to meet the Animal and Plant Health
Inspection Service requirements by its own designed methods. This would
reduce a firm's cost of research and development needed to design a
method to test immunogenicity. The proposed rule would prescribe in
vitro potency testing of serials of product with minimal costs to
manufacturing firms since the National Veterinary Services Laboratories
of the Animal and Plant Health Inspection Service have developed and
standardized the tests and would provide the critical reagents needed
to perform the tests.
The manufacturers producing E. coli bacterins would need to have
personnel proficient in conducting ELISA tests. The necessary equipment
and personnel, however, should already be available for other routine
procedures in most biologic, research and diagnostic laboratories.
Also, the ELISA in vitro potency test is less expensive than the mouse
potency test now used by the majority of firms producing E. coli
bacterins.
Under these circumstances, the Administrator of the Animal and
Plant Health Inspection Service has determined that this action would
not have a significant economic impact on a substantial number of small
entities.
Executive Order 12778
This proposed rule has been reviewed under Executive Order 12778,
Civil Justice Reform. If this proposed rule is adopted: (1) All State
and local laws and regulations that are in conflict with this rule will
be preempted; (2) no retroactive effect will be given to this rule; and
(3) administrative proceedings will not be required before parties may
file suit in court challenging this rule.
Paperwork Reduction Act
This proposed rule contains no new information collection or
recordkeeping requirements under the Paperwork Reduction Act of 1980
(44 U.S.C. 3501 et seq.).
List of Subjects in 9 CFR Part 113
Animal biologics, Export, Imports, Reporting and recordkeeping
requirements.
Accordingly, 9 CFR part 113 would be amended as follows:
PART 113--STANDARD REQUIREMENTS
1. The authority citation for part 113 would continue to read as
follows:
Authority: 21 U.S.C. 151-159; 7 CFR 2.17, 2.51, and 371.2(d).
2. A new Sec. 113.124 would be added to read as follows:
Sec. 113.124 Escherichia Coli Bacterin (E. coli).
Bacterins for the prevention of colibacillosis in mammalian
neonates born to vaccinated dams shall be prepared from bacterial
cultures which are inactivated and nontoxic. Each lot of Master Seed
Bacteria and serial or subserial of product containing Escherichia coli
shall meet the applicable requirements described in Sec. 113.100 and
the special requirements prescribed in this section. A lot of Master
Seed Bacteria found unsatisfactory by any prescribed test shall not be
used. A serial or subserial found unsatisfactory by any prescribed test
shall not be released.
(a) Each lot of Master Seed Bacteria shall be tested for
immunogenicity in each species for which the bacterin is recommended.
The immunogenicity of the Master Seed Bacteria shall be established as
follows:
(1) A Qualifying Serial of product with a minimum level of potency
shall be produced from the highest allowable passage of the Master Seed
Bacteria for use in the immunogenicity test. The allowable passage
level must be specified in the filed Outline of Production. The
relative potency of the Qualifying Serial compared to the References
shall be established as provided in paragraph (b) of this section.
(2) At least 30 pregnant cows or heifers (20 vaccinates and 10
controls) or at least 13 sows or gilts (8 vaccinates and 5 controls)
shall be used as test animals. The animals shall be randomly divided
between vaccinates and controls. If the neonates to be protected are
ovine or caprine, the number of test animals must be specified in a
protocol approved by the Animal and Plant Health Inspection Service.
(3) Pregnant dams used as vaccinates shall be injected with one
dose of the serial of product by the method recommended on the label.
If two doses are recommended, the second dose shall be administered at
the time interval recommended on the label. Serum samples shall be
collected from each dam immediately prior to each inoculation, 2 weeks
after the last inoculation, and at parturition. Colostral samples shall
be collected from each dam at parturition.
(4) Challenge of the neonates.
(i) Challenge culture(s), one for each E. coli pilus type for which
protection is claimed, shall be provided or approved by the Animal and
Plant Health Inspection Service. The challenge culture(s) used in the
efficacy test(s) shall be aliquoted into individual doses, and stored
frozen at minus 70 deg.C until needed.
(ii) After parturition, each neonate shall be allowed to suckle
normally for at least 4 hours. Four to 12 hours after parturition, each
neonate shall be weighed, have a serum sample collected, and then be
challenged.
(iii) Each neonate shall receive orally a predetermined dose of
challenge culture.
(iv) Each neonate shall be examined at least twice daily for 7 days
postchallenge for signs of colibacillosis. Neonates that die shall be
necropsied to determine the cause of death.
(5) Interpretation of results.
(i) If the mortality of the neonates from nonvaccinated bovine dams
is less than 60 percent, the test is inconclusive and may be repeated.
(ii) If at least 80 percent of the neonates from vaccinated bovine
dams do not survive without showing clinical signs of colibacillosis,
the immunogenicity of the serial of product and the Master Seed
Bacteria is unsatisfactory.
(iii) For porcine, ovine, and caprine neonates, if a statistically
significant greater number of neonates from vaccinated dams do not
survive, or the survivors do not show a significant reduction in
clinical signs of colibacillosis, or both, when compared with neonates
from nonvaccinated control dams, the immunogenicity of the serial of
product and the Master Seed Bacteria is unsatisfactory. The level of
significance required is p <0.05. clinical="" signs="" shall="" be="" evaluated="" by="" a="" method="" of="" scoring="" and="" statistical="" analysis="" acceptable="" to="" the="" animal="" and="" plant="" health="" inspection="" service.="" (b)="" references="" for="" in="" vitro="" potency="" tests.="" (1)="" comparison="" of="" the="" qualifying="" serial="" and="" working="" references="" (i)="" if="" the="" qualifying="" serial="" is="" the="" working="" reference,="" no="" comparative="" potency="" testing="" between="" the="" qualifying="" serial="" and="" the="" reference="" is="" required.="" (ii)="" if="" the="" qualifying="" serial="" is="" not="" the="" working="" reference,="" then="" the="" potency="" of="" the="" qualifying="" serial="" relative="" to="" the="" working="" reference="" shall="" be="" determined="" using="" a="" parallel="" line="" immunoassay="" that="" is="" approved="" by="" the="" animal="" and="" plant="" health="" inspection="" service.="" to="" be="" acceptable,="" the="" geometric="" mean="" of="" the="" relative="" potency="" values="" obtained="" for="" the="" qualifying="" serial="" in="" at="" least="" 5="" independent="" parallel="" line="" immunoassays="" shall="" be="" less="" than="" or="" equal="" to="" 1.00.="" (iii)="" if="" the="" master="" reference="" is="" different="" from="" either="" the="" working="" reference="" or="" the="" qualifying="" serial,="" or="" both,="" the="" dilution="" of="" the="" master="" reference="" with="" a="" potency="" equal="" to="" that="" of="" the="" working="" reference="" shall="" be="" determined="" by="" using="" a="" parallel="" line="" immunoassay="" as="" specified="" in="" paragraph="" (b)(1)(ii)="" of="" this="" section.="" (2)="" dating="" period="" for="" references.="" references="" shall="" have="" an="" initial="" dating="" period="" equal="" to="" the="" dating="" period="" of="" the="" product="" or="" as="" supported="" by="" data="" acceptable="" to="" the="" animal="" and="" plant="" health="" inspection="" service.="" the="" dating="" period="" begins="" on="" the="" date="" of="" initiation="" of="" the="" immunogenicity="" test="" or="" repeat="" immunogenicity="" test.="" the="" expiration="" date(s)="" of="" references="" shall="" be="" stated="" in="" the="" outline="" of="" production.="" (3)="" requalifying="" a="" master="" reference.="" (i)="" to="" requalify="" and="" extend="" the="" dating="" period="" of="" a="" master="" reference,="" a="" repeat="" immunogenicity="" test="" shall="" be="" conducted="" using="" a="" qualifying="" serial="" of="" product="" as="" defined="" in="" sec.="" 101.5(q)(2).="" if="" the="" master="" reference="" or="" working="" reference="" is="" a="" serial="" of="" product,="" the="" qualifying="" serial="" becomes="" the="" new="" master="" reference="" or="" the="" new="" working="" reference.="" (ii)="" the="" protocol="" and="" the="" method="" of="" evaluation="" of="" the="" repeat="" immunogenicity="" test="" to="" requalify="" a="" master="" reference="" must="" be="" approved="" by="" the="" animal="" and="" plant="" health="" inspection="" service.="" the="" results="" of="" the="" immunogenicity="" test="" specified="" in="" paragraphs="" (a)(2)="" through="" (a)(5)="" of="" this="" section="" will="" be="" a="" determining="" factor="" in="" what="" constitutes="" an="" appropriate="" test="" to="" requalify="" a="" master="" reference.="" different="" methods="" of="" requalifying="" a="" master="" reference="" are:="" (a)="" challenge="" of="" neonates="" born="" to="" vaccinated="" host="" animals.="" (b)="" challenge="" of="" vaccinated="" animals="" of="" another="" species="" whose="" immunological="" response="" has="" been="" shown="" to="" correlate="" with="" protection="" of="" neonates="" of="" the="" species="" for="" which="" the="" product="" is="" recommended.="" (c)="" serum="" and="" colostral="" antipilus="" titers="" of="" dams,="" or="" serum="" antipilus="" titers="" of="" neonates,="" or="" both,="" when="" antibody="" titers="" show="" a="" meaningful="" correlation="" to="" protection="" in="" the="" original="" immunogenicity="" test="" specified="" in="" paragraphs="" (a)(2)="" through="" (a)(5)="" of="" this="" section.="" (iii)="" requalifying="" and="" extending="" the="" dating="" period="" of="" a="" master="" reference="" may="" also="" be="" done="" by="" monitoring="" the="" potency="" of="" the="" master="" reference="" by="" in="" vitro="" methods="" over="" time="" by="" procedures="" approved="" by="" the="" animal="" and="" plant="" health="" inspection="" service="" and="" then="" conducting="" a="" repeat="" immunogenicity="" test="" as="" in="" paragraphs="" (b)(3)(i)="" and="" (b)(3)(ii)="" of="" this="" section="" when="" any="" decline="" in="" potency="" is="" detected.="" (4)="" an="" outline="" of="" production="" change="" must="" be="" approved="" by="" the="" animal="" and="" plant="" health="" inspection="" service="" to="" provide="" an="" extension="" of="" the="" expiration="" date="" of="" a="" master="" reference.="" (c)="" test="" requirements="" for="" release="" of="" serials.="" (1)="" each="" serial="" and="" subserial="" shall="" meet="" the="" applicable="" requirements="" prescribed="" in="" sec.="" 113.100="" and="" the="" requirements="" of="" this="" paragraph.="" any="" serial="" or="" subserial="" found="" unsatisfactory="" by="" a="" prescribed="" test="" shall="" not="" be="" released.="" (2)="" potency="" test.="" bulk="" or="" final="" container="" samples="" of="" completed="" product="" shall="" be="" tested="" for="" potency="" using="" the="" parallel="" line="" immunoassay="" and="" the="" working="" reference="" correlated="" directly="" to="" the="" immunogenicity="" test="" described="" in="" paragraph="" (a)="" or="" indirectly="" as="" described="" in="" paragraph="" (b)(1)="" of="" this="" section.="" the="" potency="" test(s)="" must="" be="" specific="" for="" each="" pilus="" type="" for="" which="" protection="" is="" claimed.="" the="" antigen="" capture="" antibody="" and="" the="" antigen="" indicator="" monoclonal="" antibody="" for="" the="" in="" vitro="" potency="" test="" is="" supplied="" by="" the="" animal="" and="" plant="" health="" inspection="" service.="" to="" be="" satisfactory="" and="" eligible="" for="" release,="" each="" pilus="" antigen="" in="" each="" serial="" of="" product="" shall="" have="" a="" relative="" potency="" greater="" than="" or="" equal="" to="" 1.00="" when="" compared="" to="" the="" working="" reference(s).="" serials="" not="" satisfactory="" on="" the="" initial="" relative="" potency="" test="" may="" be="" retested="" in="" accordance="" with="" sec.="" 113.8="" (c)(1)="" through="" (c)(5).="" done="" in="" washington,="" dc,="" this="" 4th="" day="" of="" october="" 1994.="" terry="" l.="" medley,="" acting="" administrator,="" animal="" and="" plant="" health="" inspection="" service.="" [fr="" doc.="" 94-25056="" filed="" 10-7-94;="" 8:45="" am]="" billing="" code="" 3410-34-m="">