[Federal Register Volume 65, Number 33 (Thursday, February 17, 2000)]
[Rules and Regulations]
[Pages 8014-8023]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 00-3832]


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DEPARTMENT OF AGRICULTURE

Animal and Plant Health Inspection Service

9 CFR Parts 145 and 147

[Docket No. 98-096-2]


National Poultry Improvement Plan and Auxiliary Provisions

AGENCY: Animal and Plant Health Inspection Service, USDA.

ACTION: Final rule.

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SUMMARY: We are amending the National Poultry Improvement Plan (the 
Plan) and its auxiliary provisions by establishing new program 
classifications and providing new or modified sampling and testing 
procedures for Plan participants and participating flocks. These 
changes were voted on and approved by the voting delegates at the 
Plan's 1998 National Plan Conference. These changes will keep the 
provisions of the Plan current with changes in the poultry industry and 
provide for the use of new sampling and testing procedures.

EFFECTIVE DATE: March 20, 2000.

FOR FURTHER INFORMATION CONTACT: Mr. Andrew R. Rhorer, Senior 
Coordinator, Poultry Improvement Staff, National Poultry Improvement 
Plan, Veterinary Services, APHIS, USDA, 1498 Klondike Road, Suite 200, 
Conyers, GA 30094-5104; (770) 922-3496.

SUPPLEMENTARY INFORMATION:

Background

    The National Poultry Improvement Plan (NPIP, also referred to below 
as

[[Page 8015]]

``the Plan'') is a cooperative Federal-State-industry mechanism for 
controlling certain poultry diseases. The Plan consists of a variety of 
programs intended to prevent and control egg-transmitted, hatchery-
disseminated poultry diseases. Participation in all Plan programs is 
voluntary, but flocks, hatcheries, and dealers must qualify as ``U.S. 
Pullorum-Typhoid Clean'' before participating in any other Plan 
program. Also, the regulations in 9 CFR part 82, subpart C, which 
provide for certain testing, restrictions on movement, and other 
restrictions on certain chickens, eggs, and other articles due to the 
presence of Salmonella enteritidis, require that no hatching eggs or 
newly hatched chicks from egg-type chicken breeding flocks may be moved 
interstate unless they are classified ``U.S. S. Enteritidis Monitored'' 
under the Plan or have met equivalent requirements for S. enteritidis 
control, in accordance with 9 CFR 145.23(d), under official 
supervision.
    The Plan identifies States, flocks, hatcheries, and dealers that 
meet certain disease control standards specified in the Plan's various 
programs. As a result, customers can buy poultry that has tested clean 
of certain diseases or that has been produced under disease-prevention 
conditions.
    The regulations in 9 CFR parts 145 and 147 (referred to below as 
the regulations) contain the provisions of the Plan. The Animal and 
Plant Health Inspection Service (APHIS) amends these provisions from 
time to time to incorporate new scientific information and technologies 
within the Plan.
    On August 10, 1999, we published in the Federal Register (64 FR 
43301-43314, Docket No. 98-096-1) a proposal to amend the regulations 
to:
    1. Establish two new classifications: ``U.S. Avian Influenza 
Clean'' for primary and multiplier egg- and meat-type breeding chicken 
flocks and ``U.S. Mycoplasma Meleagridis Clean State, Turkeys.''
    2. Identify the agar gel immunodiffusion (AGID) test and the 
enzyme-linked immunosorbent assay (ELISA) as official tests for avian 
influenza in the Plan.
    3. Allow the use of Food and Drug Administration (FDA) approved 
feed sanitizing agents or salmonella control products in certain 
chicken and turkey breeding flocks.
    4. Eliminate references to Salmonella typhimurium throughout the 
regulations.
    5. Add the colony lift assay for group D salmonella and eliminate 
the referral of all group D salmonella to APHIS' National Veterinary 
Services Laboratories (NVSL) in the laboratory protocol for isolation 
and identification of salmonella in breeding turkeys.
    6. Make several changes to the duties of the General Conference 
Committee of the NPIP.
    7. Establish technical protocol for culturing chick meconium.
    8. Provide for the use of either chick papers or meconium as 
testing samples in the ``U.S. Salmonella Monitored'' program of meat-
type breeding chickens.
    9. Amend the procedure for determining the status of a flock 
reacting to tests for Mycoplasma gallisepticum, M. synoviae, and M. 
meleagridis.
    10. Provide for the participation of emu, rhea, and cassowary 
breeding flocks in the provisions of the Plan.
    11. Remove exceptions to the requirements for pullorum typhoid 
clean States that pertain to turkey hatcheries or supply flocks.
    12. Add or amend several definitions.
    We solicited comments concerning our proposal for 60 days ending on 
October 12, 1999. We received one comment by that date. The comment was 
from a retired State animal health official. The commenter suggested 
that the 35  deg.C plate incubation temperature called for in paragraph 
(f) of proposed Sec. 147.18, ``Chick meconium testing procedure for 
salmonella,'' be changed to 37  deg.C, which is the temperature used 
for the incubation of plates in the procedure set forth in the current 
regulations in paragraph (a) of Sec. 147.11, ``Laboratory procedure 
recommended for the bacteriological examination of salmonella.'' We 
agree that the incubation temperature in Secs. 147.11(a) and 147.18(f) 
should be consistent and have made the commenter's suggested change in 
Sec. 147.18(f) of this final rule.
    Therefore, for the reasons given in the proposed rule and in this 
document, we are adopting the proposed rule as a final rule, with the 
change discussed in this document.

Executive Order 12866 and Regulatory Flexibility Act

    This 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.
    The changes contained in this document are based on the 
recommendations of representatives of member States, hatcheries, 
dealers, flockowners, and breeders who took part in the Plan's 1998 
National Plan Conference. This rule amends the Plan and its auxiliary 
provisions by establishing new program classifications and providing 
new or modified sampling and testing procedures for Plan participants 
and participating flocks. These changes, which were voted on and 
approved by the voting delegates at the Plan's 1998 National Plan 
Conference, will keep the provisions of the Plan current with changes 
in the poultry industry and provide for the use of new sampling and 
testing procedures.
    The Plan serves as a ``seal of approval'' for egg and poultry 
producers in the sense that tests and procedures recommended by the 
Plan are considered optimal for the industry. In all cases, the changes 
made by this rule have been generated by the industry itself with the 
goal of reducing disease risk and increasing product marketability. 
Because participation in the Plan is voluntary, individuals are likely 
to remain in the program as long as the costs of implementing the 
program are lower than the added benefits they receive from the 
program.
    Assuming they wished to voluntarily remain in the program, the cost 
to comply with this rule's protocols, tests, classification schemes, 
etc. will be borne primarily by the approximately 12 primary breeders 
in NPIP. However, the net economic effect of the changes on those 
breeders is expected to be positive over the long term. This is because 
the breeders' compliance costs should be more than offset by the 
expected benefits resulting from compliance (i.e., increased U.S. 
poultry exports). U.S. exports are expected to increase because, by 
serving to reduce disease risk, the protocols and procedures should 
make domestic poultry more marketable in foreign markets. That the net 
economic effect of the changes on the poultry industry is expected to 
be positive is evidenced by the fact that it was the NPIP's industry 
participants who initiated the changes.
    The precise dollar amount of the costs that the breeders will incur 
to comply with this rule is not available. However, those costs are not 
expected to be significant, especially since many of the changes are no 
more than technical corrections to the provisions of the Plan or are 
intended to bring those provisions into conformity with current 
developments in the scientific community. In 1997, the dollar value of 
U.S. exports of meat and edible offal of poultry (fresh, chilled, and 
frozen) totaled $2.2 billion (World Trade Atlas, September 1998 
edition). Even if exports increase by only 1 percent as a result of 
this rule, the benefit would be $22 million.

[[Page 8016]]

    In any event, the breeder participants in NPIP always have the 
option of withdrawing from the Plan, in which case they would not be 
subject to this rule. As indicated above, industry participation in the 
NPIP is voluntary.

Economic Effects on Small Entities

    The Regulatory Flexibility Act requires that agencies consider the 
economic effects of their rules on small entities (i.e., small 
businesses, organizations, and governmental jurisdictions). This rule 
is not expected to have a significant economic effect on a substantial 
number of small entities, if for no other reason than few, if any, of 
those entities most affected by its provisions--NPIP-participating 
breeders and producers--are small in size. The U.S. Small Business 
Administration's small entity threshold for almost all standard 
industrial classification categories for poultry and egg producers is 
annual revenues of $0.5 million or less. We believe that most, if not 
all, breeders and producers participating in the Plan generate annual 
revenues in excess $0.5 million.
    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 Catalog 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.)

Executive Order 12988

    This rule has been reviewed under Executive Order 12988, 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.

Paperwork Reduction Act

    In accordance with the Paperwork Reduction Act of 1995 (44 U.S.C. 
3501 et seq.), the information collection or recordkeeping requirements 
included in this rule have been approved by the Office of Management 
and Budget (OMB) under OMB control number 0579-0007.

List of Subjects in 9 CFR Parts 145 and 147

    Animal diseases, Poultry and poultry products, Reporting and 
recordkeeping requirements.

    Accordingly, we are amending 9 CFR parts 145 and 147 as follows:

PART 145--NATIONAL POULTRY IMPROVEMENT PLAN

    1. The authority citation for part 145 continues to read as 
follows:

    Authority: 7 U.S.C. 429; 7 CFR 2.22, 2.80, and 371.2(d).


    2. Section 145.1 is amended as follows:
    a. The definition of authorized laboratory is revised to read as 
set forth below.
    b. The definition of baby poultry is revised to read as set forth 
below.
    c. A new definition of independent flock is added, in alphabetical 
order, to read as set forth below.
    d. The definition of poultry is amended by adding the words ``emus, 
rheas, cassowaries,'' immediately after the word ``ostriches,''.
    e. The definition of S. typhimurium infection or typhimurium is 
removed.


Sec. 145.1  Definitions.

* * * * *
    Authorized laboratory. A laboratory designated by an Official State 
Agency, subject to review by the Service, to perform the blood testing 
and bacteriological examinations provided for in this part. The 
Service's review will include, but will not necessarily be limited to, 
checking records, laboratory protocol, check-test proficiency, periodic 
duplicate samples, and peer review. A satisfactory review will result 
in the authorized laboratory being recognized by the Service as a 
nationally approved laboratory qualified to perform the blood testing 
and bacteriological examinations provided for in this part.
    Baby poultry. Newly hatched poultry (chicks, poults, ducklings, 
goslings, keets, etc.).
* * * * *
    Independent flock. A flock that produces hatching eggs and that has 
no ownership affiliation with a specific hatchery.
* * * * *


Sec. 145.3  [Amended]

    3. In Sec. 145.3, the introductory text of paragraph (c) is amended 
by adding the words ``emus, rheas, cassowaries,'' immediately after the 
word ``ostriches,''.

    4. In Sec. 145.6, paragraph (e) is redesignated as paragraph (f) 
and a new paragraph (e) is added to read as follows:


Sec. 145.6  Specific provisions for participating hatcheries.

* * * * *
    (e) Any nutritive material provided to baby poultry must be free of 
the avian pathogens that are officially represented in the Plan disease 
classifications listed in Sec. 145.10.
* * * * *

    5. In Sec. 145.10, new paragraphs (r) and (s) are added to read as 
follows:


Sec. 145.10  Terminology and classification; flocks, products, and 
States.

* * * * *
    (r) U.S. Avian Influenza Clean. (See Secs. 145.23(h) and 
145.33(l).)

BILLING CODE 3410-34-P
[GRAPHIC] [TIFF OMITTED] TR17FE00.013

    (s) U.S. M. Meleagridis Clean State, Turkeys. (See Sec. 145.44(e).)

[[Page 8017]]

[GRAPHIC] [TIFF OMITTED] TR17FE00.008

BILLING CODE 3410-34-C

    6. Section 145.14 is amended as follows:
    a. In the introductory text, at the end of the first sentence, the 
words ``and ostriches blood tested under subpart F must be more than 12 
months of age'' are removed and the words ``and ostrich, emu, rhea, and 
cassowary candidates must be blood tested when at least 12 months of 
age or upon reaching sexual maturity, depending upon the species and at 
the discretion of the Official State Agency'' are added in their place.
    b. A new paragraph (d) is added to read as follows:


Sec. 145.14  Blood testing.

* * * * *
    (d) For avian influenza. The official blood tests for avian 
influenza are the agar gel immunodiffusion (AGID) test and the enzyme-
linked immunosorbent assay (ELISA).
    (1) The AGID test must be conducted on all ELISA-positive samples. 
Positive tests by AGID or ELISA must be further tested by Federal 
Reference Laboratories. Final judgment may be based upon further 
sampling or culture results.
    (2) The tests must be conducted using antigens or test kits 
approved by the Department and the Official State Agency and must be 
performed in accordance with the recommendations of the producer or 
manufacturer.
* * * * *

    7. In Sec. 145.21, the definition of chicks is revised to read as 
follows:


Sec. 145.21  Definitions.

* * * * *
    Chicks. Newly hatched chickens.
* * * * *

    8. In Sec. 145.22, a new paragraph (e) is added to read as follows:


Sec. 145.22  Participation.

* * * * *
    (e) Any nutritive material provided to chicks must be free of the 
avian pathogens that are officially represented in the Plan disease 
classifications listed in Sec. 145.10.

    9. Section 145.23 is amended as follows:
    a. In paragraph (b)(3)(i), the words ``, except turkey 
hatcheries,'' are removed.
    b. In paragraph (b)(3)(ii), the words ``, except turkey flocks,'' 
are removed.
    c. In paragraph (b)(3)(viii), the words ``, other than turkey 
flocks,'' are removed.
    d. In paragraph (b)(4), the words ``, other than turkey, waterfowl, 
exhibition poultry, and game bird supply flocks,'' are removed.

    e. Paragraph (d)(1)(ii)(B) is revised to read as follows.


Sec. 145.23  Terminology and classification; flocks and products.

* * * * *
    (d) * * *
    (1) * * *
    (ii) * * *
    (B) Mash feed may contain no animal protein other than an APPI 
animal protein product supplement manufactured in pellet form and 
crumbled: Provided, that mash feed may contain nonpelleted APPI animal 
protein product supplements if the finished feed is treated with a 
salmonella control product approved by the Food and Drug 
Administration.
* * * * *
    f. A new paragraph (h) is added to read as follows:
    (h) U.S. Avian Influenza Clean. This program is intended to be the 
basis from which the breeding-hatchery industry may conduct a program 
for the prevention and control of avian influenza. It is intended to 
determine the presence of avian influenza in breeding chickens through 
routine serological surveillance of each participating breeding flock. 
A flock and the hatching eggs and chicks produced from it will qualify 
for this classification when the Official State Agency determines that 
they have met one of the following requirements:
    (1) It is a primary breeding flock in which a minimum of 30 birds 
have been tested negative for antibodies to avian

[[Page 8018]]

influenza when more than 4 months of age. To retain this 
classification:
    (i) A sample of at least 30 birds must be tested negative at 
intervals of 90 days; or
    (ii) A sample of fewer than 30 birds may be tested, and found to be 
negative, at any one time if all pens are equally represented and a 
total of 30 birds is tested within each 90-day period.
    (2) It is a multiplier breeding flock in which a minimum of 30 
birds have been tested negative for antibodies to avian influenza when 
more than 4 months of age. To retain this classification:
    (i) A sample of at least 30 birds must be tested negative at 
intervals of 180 days; or
    (ii) A sample of fewer than 30 birds may be tested, and found to be 
negative, at any one time if all pens are equally represented and a 
total of 30 birds is tested within each 180-day period.
* * * * *
    10. In Sec. 145.31, the definition of chicks is revised to read as 
follows:


Sec. 145.31  Definitions.

* * * * *
    Chicks. Newly hatched chickens.
* * * * *
    11. In Sec. 145.32, a new paragraph (d) is added to read as 
follows:


Sec. 145.32  Participation.

* * * * *
    (d) Any nutritive material provided to chicks must be free of the 
avian pathogens that are officially represented in the Plan disease 
classifications listed in Sec. 145.10.
    12. Section 145.33 is amended as follows:
    a. In paragraph (b)(3)(i), the words ``, except turkey 
hatcheries,'' are removed.
    b. In paragraph (b)(3)(ii), the words ``, except turkey flocks,'' 
are removed.
    c. In paragraph (b)(3)(viii), the words ``, other than turkey 
flocks,'' are removed.
    d. In paragraph (b)(4), the words ``, other than turkey, waterfowl, 
exhibition poultry, and game bird supply flocks,'' are removed.
    e. In paragraph (h)(1)(ii)(A), at the end of the first sentence, 
the acronym ``(NMFS)'' is added after the word ``Service''.
    f. Paragraph (h)(1)(ii)(B) is revised to read as set forth below.
    g. Paragraph (i)(1)(vi) is amended by removing the words ``meconium 
and'' and adding the words ``meconium or'' in their place.
    h. A new paragraph (l) is added to read as follows.


Sec. 145.33  Terminology and classification; flocks and products.

* * * * *
    (h) * * *
    (1) * * *
    (ii) * * *
    (B) Mash feed may contain no animal protein other than an APPI/NMFS 
animal protein product supplement manufactured in pellet form and 
crumbled: Provided, that mash feed may contain nonpelleted APPI/NMFS 
animal protein product supplements if the finished feed is treated with 
a salmonella control product approved by the Food and Drug 
Administration.
* * * * *
    (l) U.S. Avian Influenza Clean. This program is intended to be the 
basis from which the breeding-hatchery industry may conduct a program 
for the prevention and control of avian influenza. It is intended to 
determine the presence of avian influenza in primary breeding chickens 
through routine serological surveillance of each participating breeding 
flock. A flock and the hatching eggs and chicks produced from it will 
qualify for this classification when the Official State Agency 
determines that they have met one of the following requirements:
    (1) It is a primary breeding flock in which a minimum of 30 birds 
have been tested negative for antibodies to avian influenza when more 
than 4 months of age. To retain this classification:
    (i) A sample of at least 30 birds must be tested negative at 
intervals of 90 days; or
    (ii) A sample of fewer than 30 birds may be tested, and found to be 
negative, at any one time if all pens are equally represented and a 
total of 30 birds is tested within each 90-day period.
    (2) It is a multiplier breeding flock in which a minimum of 30 
birds have been tested negative for antibodies to avian influenza when 
more than 4 months of age. To retain this classification:
    (i) A sample of at least 30 birds must be tested negative at 
intervals of 180 days; or
    (ii) A sample of fewer than 30 birds may be tested, and found to be 
negative, at any one time if all pens are equally represented and a 
total of 30 birds is tested within each 180-day period.
* * * * *
    13. In Sec. 145.41, the definition of poults is revised to read as 
follows:


Sec. 145.41   Definitions.

* * * * *
    Poults. Newly hatched turkeys.
    14. In Sec. 145.42, a new paragraph (d) is added to read as 
follows:


Sec. 145.42  Participation.

* * * * *
    (d) Any nutritive material provided to poults must be free of the 
avian pathogens that are officially represented in the Plan disease 
classifications listed in Sec. 145.10.
    15. In Sec. 145.43, paragraphs (f)(3)(ii) and (f)(3)(iii) are 
revised to read as follows:


Sec. 145.43  Terminology and classification; flocks and products.

* * * * *
    (f) * * *
    (3) * * *
    (ii) Initial feed for poults to 2 weeks of age must be manufactured 
in pellet form. Initial feed may contain no animal protein other than 
animal protein products produced under the Animal Protein Products 
Industry (APPI) Salmonella Education/Reduction Program or the Fishmeal 
Inspection Program of the National Marine Fisheries Service (NMFS). 
Finished feed must be treated with a Food and Drug Administration (FDA) 
approved salmonella control product at FDA-approved levels.
    (iii) Succeeding feed for turkeys 2 weeks or older must be either:
    (A) Pelleted feed that meets the requirements of paragraph 
(f)(3)(ii) of this section; or
    (B) Mash feed that contains no animal protein products; or
    (C) Mash feed that contains an APPI/NMFS animal protein products 
supplement that has been manufactured in pellet form and crumbled. 
Finished feed must be treated with an FDA-approved salmonella control 
product at FDA-approved levels.
* * * * *
    16. In Sec. 145.44, a new paragraph (e) is added to read as 
follows:


Sec. 145.44  Terminology and classification; States.

* * * * *
    (e) U.S. M. Meleagridis Clean State, Turkeys. (1) A State will be 
declared a U.S. M. Meleagridis Clean State, Turkeys, if the Service 
determines that:
    (i) No Mycoplasma meleagridis is known to exist nor to have existed 
in turkey breeding flocks in production within the State during the 
preceding 12 months;
    (ii) All turkey breeding flocks in production are tested and 
classified as U.S. M. Meleagridis Clean or have met equivalent 
requirements for M. meleagridis control under official supervision;
    (iii) All turkey hatcheries within the State only handle products 
that are classified as U.S. M. Meleagridis Clean or have met equivalent 
requirements for M. meleagridis control under official supervision;
    (iv) All shipments of products from turkey breeding flocks other 
than those

[[Page 8019]]

classified as U.S. M. Meleagridis Clean, or equivalent, into the State 
are prohibited;
    (v) All persons performing poultry disease diagnostic services 
within the State are required to report to the Official State Agency 
within 48 hours the source of all turkey specimens that have been 
identified as being infected with M. meleagridis;
    (vi) All reports of M. meleagridis infection in turkeys are 
promptly followed by an investigation by the Official State Agency to 
determine the origin of the infection; and
    (vii) All turkey breeding flocks found to be infected with M. 
meleagridis are quarantined until marketed under supervision of the 
Official State Agency.
    (2) The Service may revoke the State's classification as a U.S. M. 
Meleagridis Clean State, Turkeys, if any of the conditions described in 
paragraph (d)(1) of this section are discontinued. The Service will not 
revoke the State's classification as a U.S. M. Meleagridis Clean State, 
Turkeys, until it has conducted an investigation and the Official State 
Agency has been given an opportunity for a hearing in accordance with 
rules of practice adopted by the Administrator.
* * * * *
    17. In Sec. 145.52, a new paragraph (d) is added to read as 
follows:


Sec. 145.52  Participation.

* * * * *
    (d) Any nutritive material provided to baby poultry must be free of 
the avian pathogens that are officially represented in the Plan disease 
classifications listed in Sec. 145.10.


Sec. 145.53  [Amended]

    18. In Sec. 145.53, paragraph (b) is amended as follows:
    a. In paragraph (b)(3)(i), the words ``, except turkey 
hatcheries,'' are removed.
    b. In paragraph (b)(3)(ii) the words ``, except turkey flocks,'' 
are removed.
    c. In paragraph (b)(3)(viii), the words ``, other than turkey 
flocks,'' are removed.
    d. In paragraph (b)(4), the words ``, other than turkey flocks,'' 
are removed.
    19. The subpart heading for subpart F is revised to read as 
follows:

Subpart F--Special Provisions for Ostrich, Emu, Rhea, and Cassowary 
Breeding Flocks and Products

    20. In 145.61, a definition of chicks is added, in alphabetical 
order, to read as follows:


Sec. 145.61  Definitions.

* * * * *
    Chicks. Newly hatched ostriches, emus, rheas, or cassowaries.
* * * * *
    21. In Sec. 145.62, the introductory text of the section is amended 
by adding the words ``emus, rheas, and cassowaries,'' immediately after 
the word ``ostriches,'' and a new paragraph (c) is added to read as 
follows:


Sec. 145.62  Participation.

* * * * *
    (c) Any nutritive material provided to chicks must be free of the 
avian pathogens that are officially represented in the Plan disease 
classifications listed in Sec. 145.10.


Sec. 145.63  [Amended]

    22. In Sec. 145.63, paragraph (a)(2) is amended by adding the words 
``, emus, rheas, or cassowaries'' immediately after the word 
``ostriches''.

PART 147--AUXILIARY PROVISIONS ON NATIONAL POULTRY IMPROVEMENT PLAN

    23. The authority citation for part 147 continues to read as 
follows:

    Authority: 7 U.S.C. 429; 7 CFR 2.22, 2.80, and 371.2(d).


Sec. 147.4  [Removed and reserved]

    24. Section 147.4 is removed and reserved.
    25. In Sec. 147.6, paragraph (a)(14) is revised to read as follows:


Sec. 147.6  Procedure for determining the status of flocks reacting to 
tests for Mycoplasma gallisepticum, Mycoplasma synoviae, and Mycoplasma 
meleagridis.

* * * * *
    (a) * * *
    (14) If the in vivo bio-assay, PCR-based procedures, or culture 
procedures are positive, the flock will be considered infected. 
However, the following considerations may apply:
    (i) In PCR-positive flocks for which there are other negative 
mycoplasma test results, the flock's mycoplasma status should be 
confirmed through either seroconversion or culture isolation of the 
organism, or through both methods, before final determination of the 
flock's status is made.
    (ii) In flocks for which only the bio-assay is positive, additional 
in vivo bio-assay, PCR-based procedures, or cultural examinations may 
be conducted by the Official State Agency before final determination of 
the flock's status is made.
* * * * *


Secs. 147.11, 147.12, 147.14, 147.15, 147.16  [Footnotes redesignated]

    26. In Secs. 147.11, 147.12, 147.14, 147.15, 147.16, footnotes 6 
through 22 and their references are redesignated as footnotes 7 through 
23, respectively.
    27. A new Sec. 147.9 is added to read as follows:


Sec. 147.9  Standard test procedures for avian influenza.

    (a) The agar gel immunodiffusion (AGID) test should be considered 
the basic screening test for antibodies to Type A influenza viruses. 
The AGID test is used to detect circulating antibodies to Type A 
influenza group-specific antigens, namely the ribonucleoprotein (RNP) 
and matrix (M) proteins. Therefore, this test will detect antibodies to 
all influenza A viruses, regardless of subtype. The AGID test can also 
be used as a group-specific test to identify isolates as Type A 
influenza viruses. The method used is similar to that described by 
Beard. \6\ The basis for the AGID test is the concurrent migration of 
antigen and antibodies toward each other through an agar gel matrix. 
When the antigen and specific antibodies come in contact, they combine 
to form a precipitate that is trapped in the gel matrix and produces a 
visible line. The precipitin line forms where the concentration of 
antigen and antibodies is optimum. Differences in the relative 
concentration of the antigen or antibodies will shift the location of 
the line towards the well with the lowest concentration or result in 
the absence of a precipitin line. Electrolyte concentration, pH, 
temperature, and other variables also affect precipitate formation.
---------------------------------------------------------------------------

    \6\ Beard, C.W. Demonstration of type-specific influenza 
antibody in mammalian and avian sera by immunodiffusion. Bull. Wld. 
Hlth. Org. 42:779-785. 1970.
---------------------------------------------------------------------------

    (1) Materials needed.
    (i) Refrigerator (4  deg.C).
    (ii) Freezer (-20  deg.C).
    (iii) Incubator or airtight container for room temperature 
(approximately 25  deg.C) incubations.
    (iv) Autoclave.
    (v) Hot plate/stirrer and magnetic stir bar (optional).
    (vi) Vacuum pump.
    (vii) Microscope illuminator or other appropriate light source for 
viewing results.
    (viii) Immunodiffusion template cutter, seven-well pattern (a 
center well surrounded by six evenly spaced wells). Wells are 5.3 mm in 
diameter and 2.4 mm apart.
    (ix) Top loading balance (capable of measuring 0.1 gm differences).

[[Page 8020]]

    (x) Pipetting device capable of delivering 50l portions.
    (xi) Common laboratory supplies and glassware--Erlenmeyer flasks, 
graduated cylinders, pipettes, 100  x  15 mm or 60  x  15 mm petri 
dishes, flexible vacuum tubing, side-arm flask (500 mL or larger), and 
a 12- or 14-gauge blunt-ended cannula.
    (2) Reagents needed.
    (i) Phosphate buffered saline (PBS), 0.01M, pH 7.2 (NVSL media 
#30054 or equivalent).
    (ii) Agarose (Type II Medium grade, Sigma Chemical Co. Cat.# A-6877 
or equivalent).
    (iii) Avian influenza AGID antigen and positive control antiserum 
approved by the Department and the Official State Agency.
    (iv) Strong positive, weak positive, and negative control antisera 
approved by the Department and the Official State Agency (negative 
control antisera optional).
    (3) Preparing the avian influenza AGID agar.
    (i) Weigh 9 gm of agarose and 80 gm of NaCl and add to 1 liter of 
PBS (0.01 M, pH 7.2) in a 2 liter Erlenmeyer flask.
    (ii) To mix the agar, either:
    (A) Autoclave the mixture for 10 minutes and mix the contents by 
swirling after removing from the autoclave to ensure a homogeneous 
mixture of ingredients; or
    (B) Dissolve the mixture by bringing to a boil on a hot plate using 
a magnetic stir bar to mix the contents in the flask while heating. 
After boiling, allow the agar to cool at room temperature 
(approximately 25  deg.C) for 10 to 15 minutes before dispensing into 
petri plates.
    (iii) Agar can be dispensed into small quantities (daily working 
volumes) and stored in airtight containers at 4  deg.C for several 
weeks, and melted and dispensed into plates as needed.

    Note: Do not use agar if microbial contamination or precipitate 
is observed.

    (4) Performing the AGID. (i) Detection of serum antibodies.
    (A) Dispense 15 to 17 mL of melted agar into a 100  x  15 mm petri 
plate or 5 to 6 mL agar into a 60  x  15 mm petri plate using a 25 mL 
pipette. The agar thickness should be approximately 2.8 mm.
    (B) Allow plates to cool in a relatively dust-free environment with 
the lids off to permit the escape of water vapor. The lids should be 
left off for at least 15 minutes, but not longer than 30 minutes, as 
electrolyte concentration of the agar may change due to evaporation and 
adversely affect formation of precipitin lines.

    Note: Plates should be used within 24 hours after they are 
poured.

    (C) Record the sample identification, reagent lot numbers, test 
date, and identification of personnel performing and reading the test.
    (D) Using the template, cut the agar after it has hardened. Up to 
seven template patterns can be cut in a 100 x 15 mm plate and two 
patterns can be cut in a 60 x 15 mm plate.
    (E) Remove the agar plugs by aspiration with a 12- to 14-gauge 
cannula connected to a side arm flask with a piece of silicone or 
rubber tubing that is connected to a vacuum pump with tubing. Adjust 
the vacuum so that the agar surrounding the wells is not disturbed when 
removing the plugs.
    (F) To prepare the wells, either:
    (1) Place 50 l of avian influenza AGID antigen in the 
center well using a micropipette with an attached pipette tip. Place 50 
l AI AGID positive control antiserum in each of two opposite 
wells, and add 50 l per well of test sera in the four 
remaining wells. This arrangement provides a positive control line on 
one side of the test serum, thus providing for the development of lines 
of identity (see figure 1); or
    (2) Place 50 l AI AGID positive control antiserum in each 
of three alternate peripheral wells, and add 50 l per well of 
test sera in the three remaining wells. This arrangement provides a 
positive control line on each side of the test serum, thus providing 
for the development of lines of identity on both sides of each test 
serum (see figure 2).

    Note: A pattern can be included with positive, weak positive, 
and negative reference serum in the test sera wells to aid in the 
interpretation of results (see figure 3).

    (G) Cover each plate after filling all wells and allow the plates 
to incubate for 24 hours at room temperature (approximately 25  deg.C) 
in a closed chamber to prevent evaporation. Humidity should be provided 
by placing a damp paper towel in the incubation chamber. Note: 
Temperature changes during migration may lead to artifacts.
    (ii) Interpretation of test results.
    (A) Remove the lid and examine reactions from above by placing the 
plate(s) over a black background, and illuminate the plate with a light 
source directed at an angle from below. A microscope illuminator works 
well and allows for varying intensities of light and positions.
    (B) The type of reaction will vary with the concentration of 
antibody in the sample being tested. The positive control serum line is 
the basis for reading the test. If the line is not distinct, the test 
is not valid and must be repeated. The following types of reactions are 
observed (see figure 3):
    (1) Negative reaction. The control lines continue into the test 
sample well without bending or with a slight bend away from the antigen 
well and toward the positive control serum well.
    (2) Positive reaction. The control lines join with, and form a 
continuous line (line of identity) with, the line between the test 
serum and antigen. The location of the line will depend on the 
concentration of antibodies in the test serum. Weakly positive samples 
may not produce a complete line between the antigen and test serum but 
may only cause the tip or end of the control line to bend inward toward 
the test well.
    (3) Non-specific lines. These lines occasionally are observed 
between the antigen and test serum well. The control lines will pass 
through the non-specific line and continue on into the test serum well. 
The non-specific line does not form a continuous line with positive 
control lines.

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    (b) The enzyme-linked immunosorbent assay (ELISA) may be used as a 
screening test for avian influenza. Use only federally licensed ELISA 
kits and follow the manufacturer's instructions. All ELISA-positive 
serum samples must be confirmed with the AGID test conducted in 
accordance with paragraph (a) of this section.


Sec. 147.11  [Amended]

    28. Section 147.11 is amended as follows:
    a. In paragraph (b)(2)(iii) the words ``A group D colony lift assay 
may be utilized to signal the presence of the hard-to-detect group D 
salmonella colonies on agar culture plates.'' are added after the final 
sentence.
    b. In paragraph (b)(2)(v), the words ``at the National Veterinary 
Services Laboratory'' are removed.
    29. A new Sec. 147.18 is added to read as follows:


Sec. 147.18  Chick meconium testing procedure for salmonella.

    Procedure:
    (a) Record the date, source, and flock destination on the 
``Meconium Worksheet.''
    (b) Shake each plastic bag of meconium until a uniform consistency 
is achieved.
    (c) Transfer a 25 gm sample of meconium to a sterile container. Add 
225 mL of a preenrichment broth to each sample (this is a 1:10 
dilution), mix gently, and incubate at 37  deg.C for 18-24 hours.
    (d) Enrich the sample with selective enrichment broth for 24 hours 
at 42  deg.C.
    (e) Streak the enriched sample onto brilliant green-Novobiocin 
(BGN) agar and xylose-lysine-tergitol 4 (XLT4) agar.
    (f) Incubate both plates at 37  deg.C for 24 hours and process 
suspect salmonella colonies according to Sec. 147.11.
    30. In Sec. 147.43, paragraphs (d)(1) through (d)(4) are 
redesignated as paragraphs (d)(3) through (d)(6), respectively, and new 
paragraphs (d)(1), (d)(2), (d)(7), and (d)(8) are added to read as 
follows:


Sec. 147.43  General Conference Committee.

* * * * *
    (d) * * *
    (1) Advise and make recommendations to the Department on the 
relative importance of maintaining, at all times, adequate departmental 
funding for the NPIP to enable the Senior Coordinator and staff to 
fully administer the provisions of the Plan.
    (2) Advise and make yearly recommendations to the Department with 
respect to the NPIP budget well in advance of the start of the 
budgetary process.
* * * * *
    (7) Serve as a direct liaison between the NPIP and the United 
States Animal Health Association.
    (8) Advise and make recommendations to the Department regarding 
NPIP involvement or representation at poultry industry functions and 
activities as deemed necessary or advisable for the purposes of the 
NPIP.


Sec. 147.45  [Amended]

    31. Section 147.45 is amended by removing the words ``and E'' and 
adding the words ``E, and F'' in their place.
    32. In Sec. 147.46, the introductory text of paragraph (a) is 
amended by removing the word ``four'' and adding the word ``five'' in 
its place, and a new paragraph (a)(5) is added to read as follows:


Sec. 147.46  Committee consideration of proposed changes.

    (a) * * *
    (5) Ostriches, emus, rheas, and cassowaries.
* * * * *

    Done in Washington, DC, this 11th day of February 2000.
Bobby R. Acord,
Acting Administrator, Animal and Plant Health Inspection Service.
[FR Doc. 00-3832 Filed 2-16-00; 8:45 am]
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