National Poultry Improvement Plan and Auxiliary Provisions

DEPARTMENT OF AGRICULTURE

Animal and Plant Health Inspection Service

9 CFR Parts 145 and 147

[Docket No. 98-096-1]

National Poultry Improvement Plan and Auxiliary Provisions

AGENCY: Animal and Plant Health Inspection Service, USDA.

ACTION: Proposed rule.

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SUMMARY: We are proposing to amend 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. The

proposed changes were voted on and approved by the voting delegates at

the Plan's 1998 National Plan Conference. These changes would keep the

provisions of the Plan current with changes in the poultry industry and

provide for the use of new sampling and testing procedures.

DATES: We invite you to comment on this docket. We will consider all

comments that we receive by October 12, 1999.

ADDRESSES: Please send your comment and three copies to: Docket No. 98-

096-1, Regulatory Analysis and Development, PPD, APHIS, Suite 3C03 4700

River Road, Unit 118, Riverdale, MD 20737-1238. Please state that your

comment refers to Docket No. 98-096-1.

You may read any comments that we receive on this docket in our

reading room. The reading room is located in room 1141 of the USDA

South Building, 14th Street and Independence Avenue, SW., Washington,

DC. Normal reading room hours are 8 a.m. to 4:30 p.m., Monday through

Friday, except holidays. To be sure someone is there to help you,

please call (202) 690-2817 before coming.

APHIS documents published in the Federal Register, and related

information, including the names of organizations and individuals who

have commented on APHIS rules, are available on the Internet at http://

www.aphis.usda.gov/ppd/rad/webrepor.html.

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.

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SUPPLEMENTARY INFORMATION:

Background

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

as ``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. In this document, we are proposing 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.

These proposed amendments are consistent with the recommendations

approved by the voting delegates to the National Plan Conference that

was held from July 15 to 17, 1998.

Participants in the 1998 National Plan Conferences represented

flockowners, breeders, hatcherymen, and Official State Agencies from

all cooperating States. The proposed amendments are discussed in

greater detail below.

U.S. Avian Influenza Clean

We are proposing to add a new Sec. 145.23(h) to establish a new

``U.S. Avian Influenza Clean'' classification for egg-type chickens and

meat-type chickens. This proposed program is intended to be the basis

from which the breeding-hatchery industry could conduct a program for

the prevention and control of avian influenza. The program would enable

flockowners 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 would qualify for this proposed classification when the Official

State Agency determined that they have met the qualifying requirements.

For primary breeding flocks, a minimum of 30 birds would have to

have been tested negative for antibodies to avian influenza when the

flock is more than 4 months of age to qualify for the classification.

After qualifying, a sample of at least 30 birds from the flock would

have to be tested negative at intervals of 90 days to retain the

classification. As noted above, this routine serological surveillance

would allow flockowners to monitor their flocks for the presence of

avian influenza. Under the proposed classification criteria,

flockowners could test samples of fewer than 30 birds at any one time

if all pens were equally represented and a total of 30 birds was tested

within each 90-day period. This would provide an alternative for

flockowners who may find it easier to spread the necessary testing out

over a period of time rather than testing all the birds at the same

time.

The qualifying requirements for multiplier breeding flocks would be

the same as for primary breeding flocks with one exception: Instead of

having to test a sample of 30 birds every 90 days to retain the

classification, the testing interval for multiplier breeding flocks

would be 30 birds every 180 days. This longer testing interval for

multiplier breeding flocks is used throughout the Plan in other disease

classifications and is appropriate because there are many more

multiplier breeding flocks than primary breeding flocks--the ratio is

roughly 5\1/2\ to 1. With the much larger number of multiplier breeding

flocks, it works out that multiplier breeding flocks would actually be

tested nearly three times more often during the course of a year than

the primary breeding flocks in a given State. Given that the multiplier

breeding flocks are held in comparatively closer proximity and looser

biosecurity conditions, relative to the primary breeding flocks, the

health status of one multiplier flock is considered a reliable

indicator of the health status of the surrounding multiplier flocks.

This is especially true with regard to avian influenza, given the fact

that the level of avian influenza infection in the flocks in an area

where the disease is present would be very high, if not 100 percent.

Given these considerations, we believe that this longer interval for

testing multiplier breeding flocks would provide an appropriate level

of surveillance for avian influeza.

U.S. M. Meleagridis Clean State, Turkeys

We are proposing to add a new Sec. 145.44(e) to establish a new

``U.S. M. Meleagridis Clean State'' classification for turkeys. This

proposed new classification would be given to qualifying States in

which all turkey flocks have been shown to be free of Mycoplasma

meleagridis and in which no M. meleagridis has been detected in

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turkey flocks for at least the previous 12 months.

For a State to qualify for this proposed new classification, all

turkey breeding flocks in production in the State would have to qualify

as ``U.S. M. Meleagridis Clean'' or its equivalent, and all turkey

hatcheries within the State would have to handle only products that are

classified as ``U.S. M. Meleagridis Clean'' or its equivalent.

Additionally, all shipments of products from turkey breeding flocks

other than those classified as ``U.S. M. Meleagridis Clean'' or its

equivalent into the State would be prohibited.

All persons performing poultry disease diagnostic services within

the State would be required to report to the Official State Agency

within 48 hours the source of all turkey specimens that are identified

as being infected with M. meleagridis; such reports would have to be

followed by an investigation by the Official State Agency to determine

the origin of the infection. Any turkey breeding flock found to be

infected with M. meleagridis would have to be quarantined until

marketed under supervision of the Official State Agency.

If a State no longer met any of the above conditions, or if

repeated outbreaks of M. meleagridis occurred in turkey breeding

flocks, or if an infection spread from the premises on which it

originated, APHIS would have grounds to revoke its determination that

the State was entitled to the classification. Such action would not be

taken until APHIS had conducted a thorough investigation and the

Official State Agency had been given an opportunity for a hearing in

accordance with rules of practice adopted by the Administrator.

Tests for Avian Influenza

We are proposing to amend Sec. 145.14, ``Blood testing,'' to

designate the agar gel immunodiffusion (AGID) test and the enzyme-

linked immunosorbent assay (ELISA) as the official Plan blood tests for

avian influenza. These tests would have to be conducted using antigens

or test kits approved by the Department and the Official State Agency

and would have to be performed in accordance with the recommendations

and instructions provided by the test's producer or manufacturer. These

proposed requirements would ensure that the tests are routinely

conducted in a consistent and accurate manner. We would allow the use

of either test because some laboratories find the ELISA a less labor-

intensive test to perform, but the AGID is recognized by the Office of

International Epizootics as the international standard test for avian

influenza. We would require, however, that any ELISA positive tests

would have to be check tested using the AGID, since the AGID test is

specifically required by many of the countries to which the United

States poultry industry exports its products.

The instructions for conducting the AGID and ELISA tests would be

set out in a new Sec. 147.9. Paragraph (a) of the proposed new section

would provide detailed instructions regarding the use of AGID test as a

screening test for avian influenza, including lists of the materials

and reagents needed for the test and directions for preparing the avian

influenza AGID agar, performing the AGID test, and interpreting test

results. Paragraph (b) of the proposed new section would explain that

the ELISA may also be used as a screening test for avian influenza and

would require the use of federally licensed ELISA kits in accordance

with the manufacturer's instructions. The AGID testing protocols, which

are set out in Sec. 147.9 in the rule portion of this document, were

developed by NVSL and have been reviewed by avian influenza technical

experts. Because proposed Sec. 147.9 contains a footnote, we would also

renumber the remaining footnotes in part 147 to accommodate its

inclusion.

Feed and Salmonella Control Products

The definitions of baby poultry in Sec. 145.1, chicks in

Secs. 145.21 and 145.31, and poults in Sec. 145.41 all refer to newly

hatched birds that have not been fed or watered. The limitation on

feeding and watering can be traced back to the standard practices for

shipping mail order chicks and poults that were developed when it was

impractical to include food or water in the chick or poult boxes. Now,

however, gels are available that can easily be placed in chick and

poult boxes. The use of these gels has become widespread in the

industry and has virtually eliminated primary mortality in baby poultry

due to dehydration. Therefore, we are proposing to amend the

definitions of baby poultry, chicks, and poults to remove the words

``that have not been fed or watered'' in order for the regulations in

part 145 to reflect actual poultry industry practice.

We do believe, however, that it is important to ensure that the

gels or other nutrients provided to the baby poultry in participating

flocks and hatcheries do not expose the chicks or poults to any of the

diseases addressed by Plan programs. Accordingly, we are proposing to

add a paragraph to each of the subparts in part 145 to inform Plan

participants that any nutritive material provided to baby poultry must

be free of the avian pathogens that are officially represented in Plan

disease classifications, which are listed in Sec. 145.10. This

paragraph would be added to Sec. 145.6, ``Specific provisions for

participating hatcheries,'' in subpart A and to the ``Participation''

sections (i.e., Secs. 145.21, 145.31, 145.41, 145.51, and 145.61) of

the other five subparts.

We are also proposing to amend Secs. 145.23(d), 145.33(h), and

145.43(f) to provide for the use of FDA-approved salmonella control

products on finished feed as an additional measure for reducing

salmonella in breeding flocks. The Plan's provisions currently provide

for the use of feed with no animal protein or require feed containing

animal protein to meet specified requirements. Allowing salmonella

control products that have been approved by the FDA to be used in

poultry feed would provide flockowners with an alternative means of

reducing the likelihood of salmonella being introduced into their

breeding flocks through feed.

Addition of Emus, Rheas, and Cassowaries

We are proposing to amend parts 145 and 147 to provide for the

participation of emu, rhea, and cassowary breeding flocks in the

provisions of the Plan. The proposed addition to the Plan of provisions

for emu, rhea, and cassowary breeding flocks was voted on and approved

by the voting delegates at the Plan's 1998 National Plan Conference and

follows the addition in 1998 of provisions for the participation of

ostrich breeding flocks. Adding provisions to the Plan for emu, rhea,

and cassowary breeding flocks would make it possible for the owners of

those flocks to voluntarily participate in the Plan's programs for the

prevention and control of egg-transmitted, hatchery-disseminated

poultry diseases. To integrate emus, rheas, and cassowaries into the

provisions of the Plan, we are proposing to amend several sections of

the regulations.

First, we would add emus, rheas, and cassowaries to the definition

of poultry in Sec. 145.1 to ensure that the general provisions of the

regulations would apply, where applicable, to emus, rheas, and

cassowaries as well as to the types of poultry already covered by the

Plan. With the proposed addition of emus, rheas, and cassowaries, the

definition of poultry would read: ``Domesticated fowl, including

chickens, turkeys, ostriches, emus, rheas, and cassowaries, waterfowl,

and game birds, except doves and pigeons, which are bred for the

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primary purpose of producing eggs or meat.''

Under Sec. 145.3(c), ``Participation,'' a Plan participant in any

State must participate with all of his poultry hatching egg supply

flocks and hatchery operations in that State. To demonstrate compliance

with that requirement, the Plan participant must submit a report of

each of his breeding flocks within the State to the Official State

Agency before the birds in a breeding flock reach 24 weeks of age or,

in the case of ostriches, before the birds reach 20 months of age.

Under the provisions of this proposed rule, those participation

requirements would also apply to emu, rhea, and cassowary hatching egg

supply flocks and hatchery operations. Because emus, rheas, and

cassowaries mature at a rate comparable to that of ostriches, a

participant would have to report his or her emu, rhea, or cassowary

breeding flocks to the Official State Agency before the birds in the

flock reach 20 months of age, as is the case for ostriches, rather than

24 weeks of age as required for other poultry.

We would amend the introductory text of Sec. 145.14 by adding a

provision regarding the blood testing of emus, rheas, and cassowaries.

That text currently states that poultry must be more than 4 months of

age when blood tested for an official classification, except for

turkeys, which may be blood tested at 12 weeks of age; game birds,

which may be blood tested when more than 4 months of age or upon

reaching sexual maturity, whichever comes first; and ostriches, which

must be more than 12 months of age.

In providing for the blood testing of emus, rheas, and cassowaries,

we are also proposing to amend the exception regarding ostriches.

Specifically, we would provide that ostrich, emu, rhea, and cassowary

candidates would 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. (As noted in the previous

paragraph, ostriches currently must be ``more than 12 months of age''

when blood tested.) We would provide for blood testing to occur when

the birds are at least 12 months of age or upon reaching sexual

maturity because these four species will not reach sexual maturity at

the same age, although approximately a year after hatching is an

appropriate general time frame. The immature birds are kept in a

juvenile rearing facility for about a year after hatching, so it would

not be necessary to test them for an official classification until such

time as they were ready to be integrated into a breeding flock.

The special provisions for emu, rhea, and cassowary breeding flocks

would be added to subpart F (Secs. 145.61 through 145.63), which

currently pertains only to ostriches. To include emus, rheas, and

cassowaries in subpart F, we would add the words ``emu, rhea, and

cassowary'' after the word ``ostrich'' in the following places:

The title of the subpart. As amended, the title would read

``Special Provisions for Ostrich, Emu, Rhea, and Cassowary Breeding

Flocks.''

The introductory text of Sec. 145.62. Emus, rheas, and cassowaries

would be subject to the section's requirement that participating

flocks, and the eggs and chicks produced from them, must comply with

the applicable general provisions of subpart A and the special

provisions of subpart F.

Paragraph (a) of Sec. 145.62. Emus, rheas, and cassowaries would

lose their identity under Plan terminology--that is, they would not be

considered U.S. Pullorum-Typhoid Clean poultry--if they were not

maintained under the conditions prescribed in Sec. 145.5(a). Under

Sec. 145.5(a), poultry equipment, poultry houses, and the land in their

immediate vicinity must be kept in sanitary condition, and the

participating flock, its eggs, and all equipment used in connection

with the flock must be kept separated from nonparticipating flocks. The

sanitation and segregation described in Sec. 145.5(a) are important

factors in maintaining the health of flocks, which is why we would

require that those conditions be met in order for started poultry to

retain its identity under Plan terminology.

Paragraph (b) of Sec. 145.62. The hatching eggs produced by emu,

rhea, and cassowary primary breeding flocks would have to be fumigated

or otherwise sanitized; that paragraph also refers the reader to

Sec. 147.22, which contains procedures for the sanitation of hatching

eggs. This proposed requirement for the sanitation of hatching eggs

would serve to help prevent the transmission of egg-disseminated

diseases that could be spread by unsanitized eggs.

Paragraph (a) of Sec. 145.63. Emu, rhea, and cassowary flocks would

be subject to the same qualifying criteria for the U.S. Pullorum-

Typhoid Clean classification as are ostrich flocks. Emu, rhea, and

cassowary flocks seeking the U.S. Pullorum Typhoid Clean classification

would have to demonstrate their freedom from pullorum and typhoid to

the Official State Agency through annual blood testing or a

bacteriological monitoring program.

The regulations in Sec. 147.45 regarding official delegates to Plan

conferences refer to the programs prescribed in subparts B, C, D, and E

of part 145. Similarly, the regulations in Sec. 147.46 refer to four

committees within the Plan (egg-type chickens, meat-type chickens,

turkeys, and waterfowl, exhibition poultry, and game birds) that have

been established to consider possible changes to the Plan's provisions.

In order to fully integrate ostrich, emu, rhea, and cassowary flocks

into the Plan and provide for the full participation of their

flockowners, we are proposing to amend Sec. 147.45 so that it refers to

subpart F and Sec. 147.46 so that it refers to a committee for

ostriches, emus, rheas, and cassowaries.

Mycoplasma Status of Flocks

In Sec. 147.6, ``Procedure for determining the status of flocks

reacting to tests for Mycoplasma gallisepticum, Mycoplasma synoviae,

and Mycoplasma meleagridis,'' paragraph (a)(14) currently provides that

a flock will be considered infected with mycoplasma based on the

results of an in vivo bio-assay, polymerase chain reaction (PCR) based

procedures, or cultural examinations. That paragraph does, however,

provide that if only the bio-assay is positive, additional in vivo bio-

assays, PCR-based procedures, or cultural examinations may be conducted

by the Official State Agency before a final determination on the

flock's mycoplasma status is made. In this document, we are proposing

to amend that paragraph to provide the same opportunity for additional

testing in instances when only the results of the PCR-based procedure

are positive. This proposed change would allow Official State Agencies

to corroborate the findings of the PCR-based procedures through the use

of seroconversion or culture isolation of the mycoplasma organism.

Colony Lift Assay

We are proposing to amend Sec. 147.11(b), which contains

bacteriological examination procedures for use with turkey specimens

and environmental specimens from turkey flocks, to provide for the use

of the colony lift assay as a means for laboratories to pick group D

salmonella colonies from selective and non-selective agar culture

plates. Group D salmonella colonies are difficult to detect on agar

culture plates, so allowing the use of a group D colony lift assay

would increase the sensitivity of the culture procedure by eliminating

the randomness of selecting colonies, as the

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randomness could lead to group D cultures being missed on the agar

plate.

We are also proposing to amend the turkey culturing provisions in

Sec. 147.11(b) to remove the requirement that all salmonella group D

cultures be referred to NVSL for serotyping. Authorized laboratories

are capable of conducting the serotyping themselves, so there is no

need for the cultures to be referred to NVSL. These proposed changes

would make the turkey culturing requirements consistent with the

corresponding requirements for egg-type and meat-type chickens.

Chick Meconium Testing Procedure

We are proposing to add a new Sec. 147.18 to provide a testing

procedure for chick meconium. This procedure, which is set out in the

rule portion of this document, would be added because the ``U.S.

Salmonella Monitored'' classification requires the testing of chick

meconium. Because the testing is required by the Plan, it is necessary

to provide an official procedure for the collection of samples and

laboratory testing. The testing protocol was developed by scientists

from the Primary Poultry Breeders Veterinarian Roundtable who have

expertise in salmonella isolation and identification.

General Conference Committee

Section 147.43 explains the membership, duties, and functions of

the Plan's General Conference Committee (GCC), which is the body that

provides advice and assistance to the Department in its administration

of the NPIP. At the 1998 National Plan Conference, the voting delegates

approved additional duties that the Plan membership wishes the GCC to

undertake. Those additional duties are:

Advise and make recommendations to the Department to the

relative importance of maintaining, at all times, adequate Department

funding for the NPIP to enable the Senior Coordinator and staff to

fully administer the provisions of the Plan.

Advise and make yearly recommendations to the Department

with respect to the NPIP budget well in advance of the start of the

budgetary process.

Serve as a direct liaison between the NPIP and the United

States Animal Health Association.

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.

We are, therefore, proposing to amend Sec. 147.43 to reflect these

additional advisory and liaison duties.

Definitions

In Sec. 145.1, we are proposing to amend the definition of

authorized laboratory and to add a definition of independent flock. The

definition of authorized laboratory currently reads: ``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.'' We are proposing to add to the end of that

definition the following: ``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.'' Authorized laboratories have developed into a

significant component of the Plan, and the types of tests that are

conducted by authorized laboratories on behalf of the NPIP have become

more varied in recent years as the Plan has become involved in the

certification of essentially all of the live poultry and poultry meat

products produced in the United States. The delegates at the Plan's

1998 National Plan Conference voted to add the specific review elements

described above to the definition of authorized laboratory in order to

provide for uniformity and consistency among the Plan's 125 authorized

laboratories.

There are three categories of participation in the NPIP:

Hatcheries, independent flocks, and dealers. Hatcheries and dealers are

already addressed in Sec. 145.1, but there is not currently a

definition of the term ``independent flock.'' Therefore, we are

proposing to add the following definition of independent flock to

Sec. 145.1: ``A flock that produces hatching eggs and that has no

ownership affiliation with a specific hatchery.''

We are also proposing to amend Sec. 145.61, which provides

definitions for the specific provisions of subpart F. That section does

not currently include a definition for the term ``chick,'' which is

used several times in that subpart. Therefore, we are also proposing to

amend Sec. 145.61 to add a definition of chick, which would read

``Newly hatched ostriches, emus, rheas, or cassowaries.'' Adding this

definition, which is consistent with the definition provided for the

same term in the other four subparts of part 145, would clarify what is

intended when the term ``chick'' is used in subpart F.

Miscellaneous

Prior to 1970, the provisions of the regulations that apply to

turkeys were not part of the NPIP, but were instead part of the

National Turkey Improvement Plan (NTIP). Because turkeys were not

included in the NPIP, the NPIP regulations specifically excluded turkey

hatcheries, hatchery supply flocks, and breeding flocks from the

criteria used to determine the pullorum-typhoid status of meat-type and

egg-type chicken breeding flocks and waterfowl, exhibition poultry, and

game bird breeding flocks. When the NTIP was integrated into the NPIP,

those exemptions should have been removed from the regulations but were

not, which has resulted in a discrepancy between the U.S. Pullorum-

Typhoid Clean classification criteria for turkeys and the same criteria

for chickens and waterfowl, exhibition poultry, and game birds. A

similar discrepancy exists between the U.S. Pullorum-Typhoid Clean

classification criteria for egg- and meat-type chicken supply flocks

and the requirements for waterfowl, exhibition poultry, and game bird

supply flocks. In order to eliminate those discrepancies, we are

proposing to amend Secs. 145.23, 145.33, and 145.53 to eliminate the

incorrect exemptions discussed in this paragraph.

We are also proposing to amend Sec. 145.1 to remove the definition

of S. typhimurium infection or typhimurium because the disease is not

referred to, nor is the term itself used, in part 145. Further, because

the Plan does not include any programs for the prevention or control of

Salmonella typhimurium, the instructions provided in Sec. 147.4, ``The

tube agglutination test for S. typhimurium,'' are unnecessary.

Therefore, we are proposing to remove Sec. 147.4 from the regulations.

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.

The proposed 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. The proposed changes would amend the Plan and

its auxiliary

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provisions by establishing new program classifications and providing

new or modified sampling and testing procedures for Plan participants

and participating flocks. The proposed changes were voted on and

approved by the voting delegates at the Plan's 1998 National Plan

Conference. These changes would 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

proposed in this document 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 the proposed protocols, tests, classification schemes,

etc. would be borne primarily by the approximately 12 primary breeders

in NPIP. However, the net economic effect of the proposed 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 proposed protocols and procedures

should make domestic poultry more marketable in foreign markets. That

the net economic effect of the proposed changes on the poultry industry

is expected to be positive is evidenced by the fact the industry

participants of NPIP themselves initiated the proposed changes.

The precise dollar amount of the costs that the breeders would

incur to comply with the proposed changes is not available. However,

those costs are not expected to be significant, especially since many

of the proposed 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 increased by only 1

percent as a result of the proposed changes, the benefit would be $22

million.

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 the proposed changes. 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 its rules on small entities, i.e., small

businesses, organizations, and governmental jurisdictions. The changes

proposed in this document are 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 the

proposed changes--i.e., 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 would

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 proposed rule has been reviewed under Executive Order 12988,

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

In accordance with section 3507(d) of the Paperwork Reduction Act

of 1995 (44 U.S.C. 3501 et seq.), the information collection or

recordkeeping requirements included in this proposed rule have been

submitted for approval to the Office of Management and Budget (OMB).

Please send written comments to the Office of Information and

Regulatory Affairs, OMB, Attention: Desk Officer for APHIS, Washington,

DC 20503. Please state that your comments refer to Docket No. 98-096-1.

Please send a copy of your comments to: (1) Docket No. 98-096-1,

Regulatory Analysis and Development, PPD, APHIS, suite 3C03, 4700 River

Road, Unit 118, Riverdale, MD 20737-1238, and (2) Clearance Officer,

OCIO, USDA, room 404-W, 14th Street and Independence Avenue, SW.,

Washington, DC 20250. A comment to OMB is best assured of having its

full effect if OMB receives it within 30 days of publication of this

proposed rule.

The NPIP is a voluntary Federal-State-industry mechanism for

controlling certain poultry diseases and for improving poultry breeding

flocks and products through disease control techniques. APHIS is

responsible for administering the Plan, the primary purpose of which is

to protect the health of the U.S. poultry population.

This proposed rule would, among other things, amend the provisions

of the Plan to provide for the participation of emu, rhea, and

cassowary breeding flocks in the Plan. This would make it possible for

the owners of these breeding flocks to voluntarily participate in the

NPIP's programs for the prevention and control of egg-transmitted,

hatchery-disseminated poultry diseases. Including emu, rhea, and

cassowary in the provisions of the Plan would enhance our ability to

protect the United States against certain poultry diseases.

Our proposed rule would also establish a new ``U.S. M. Meleagridis

Clean State'' classification for turkeys that would be awarded to

qualifying States in which all turkey flocks have been shown to be free

of this disease. Achieving this classification would enhance the value

of turkey products in national and international trade, and would

provide flock owners with added incentive to eliminate this disease

from their flocks.

Expanding the Plan to include emu, rhea, and cassowary breeding

flocks and establishing a ``U.S. M. Meleagridis Clean State''

classification for turkeys will necessitate the use of two information

collection activities that will (1) alert us to the disease status of

turkeys in any given State and (2) alert us when any given owner of

emu, rhea, or cassowary flocks opts to enroll these flocks in the Plan.

We are asking OMB to approve our use of these information collection

activities, which are a necessary element of the Plan's

[[Page 43307]]

programs to prevent the spread of contagious poultry diseases within

the United States.

We are soliciting comments from the public (as well as affected

agencies) concerning our proposed information collection and

recordkeeping requirements. These comments will help us:

(1) Evaluate whether the proposed information collection is

necessary for the proper performance of our agency's functions,

including whether the information will have practical utility;

(2) Evaluate the accuracy of our estimate of the burden of the

proposed information collection, including the validity of the

methodology and assumptions used;

(3) Enhance the quality, utility, and clarity of the information to

be collected; and

(4) Minimize the burden of the information collection on those who

are to respond (such as through the use of appropriate automated,

electronic, mechanical, or other technological collection techniques or

other forms of information technology, e.g., permitting electronic

submission of responses).

Estimate of burden: Public reporting burden for this collection of

information is estimated to average 0.2 hours per response.

Respondents: Flock owners, breeders, hatchery operators, and State

veterinary medical officers.

Estimated annual number of respondents: 10.

Estimated annual number of responses per respondent: 1.

Estimated annual number of responses: 10.

Estimated total annual burden on respondents: 2 hours.

Copies of this information collection can be obtained from:

Clearance Officer, OCIO, USDA, room 404-W, 14th Street and Independence

Avenue, SW., Washington, DC 20250.

List of Subjects in 9 CFR Parts 145 and 147

Animal diseases, Poultry and poultry products, Reporting and

recordkeeping requirements.

Accordingly, we are proposing to amend 9 CFR parts 145 and 147 as

follows:

PART 145--NATIONAL POULTRY IMPROVEMENT PLAN

1. The authority citation for part 145 would continue to read as

follows:

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

2. Section 145.1 would be amended as follows:

a. The definition of authorized laboratory would be revised to read

as set forth below.

b. The definition of baby poultry would be revised to read as set

forth below.

c. A new definition of independent flock would be added, in

alphabetical order, to read as set forth below.

d. The definition of poultry would be amended by adding the words

``emus, rheas, cassowaries,'' immediately after the word

``ostriches,''.

e. The definition of S. typhimurium infection or typhimurium would

be 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) would be

amended by adding the words ``emus, rheas, cassowaries,'' immediately

after the word ``ostriches,''.

4. In Sec. 145.6, paragraph (e) would be redesignated as paragraph

(f) and a new paragraph (e) would be 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) would be 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-U

[GRAPHIC] [TIFF OMITTED] TP10AU99.018

[[Page 43308]]

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

[GRAPHIC] [TIFF OMITTED] TP10AU99.019

BILLING CODE 3410-34-C

6. Section 145.14 would be 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'' would be 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'' would be

added in their place.

b. A new paragraph (d) would be 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 or 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 would be revised to

read as follows:

Sec. 145.21 Definitions.

* * * * *

Chicks. Newly hatched chickens.

* * * * *

8. In Sec. 145.22, a new paragraph (e) would be 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 would be amended as follows:

a. In paragraph (b)(3)(i), the words ``, except turkey

hatcheries,'' would be removed.

b. In paragraph (b)(3)(ii), the words ``, except turkey flocks,''

would be removed.

c. In paragraph (b)(3)(viii), the words ``, other than turkey

flocks,'' would be removed.

d. In paragraph (b)(4), the words ``, other than turkey, waterfowl,

exhibition poultry, and game bird supply flocks,'' would be removed.

e. Paragraph (d)(1)(ii)(B) would be revised.

f. A new paragraph (h) would be added 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 non-pelleted APPI animal

protein product supplements if the finished feed is treated with a

salmonella control product approved by the Food and Drug

Administration.

* * * * *

(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

[[Page 43309]]

determines that they have met one of the following requirements:

(1) It is a primary breeding flock in which 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 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 would be revised to

read as follows:

Sec. 145.31 Definitions.

* * * * *

Chicks. Newly hatched chickens.

* * * * *

11. In Sec. 145.32, a new paragraph (d) would be 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 would be amended as follows:

a. In paragraph (b)(3)(i), the words

``, except turkey hatcheries,'' would be removed.

b. In paragraph (b)(3)(ii), the words

``, except turkey flocks,'' would be removed.

c. In paragraph (b)(3)(viii), the words ``, other than turkey

flocks,'' would be removed.

d. In paragraph (b)(4), the words

``, other than turkey, waterfowl, exhibition poultry, and game bird

supply flocks,'' would be removed.

e. Paragraph (h)(1)(ii)(B) would be revised.

f. Paragraph (i)(1)(vi) would be amended by removing the words

``meconium and'' and adding the words ``meconium or'' in their place.

g. A new paragraph (l) would be 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 non-pelleted 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 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 would be revised to

read as follows:

Sec. 145.41 Definitions.

* * * * *

Poults. Newly hatched turkeys.

14. In Sec. 145.42, a new paragraph (d) would be 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) would be

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) would be 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

[[Page 43310]]

or have met equivalent requirements for M. meleagridis control under

official supervision;

(iv) All shipments of products from turkey breeding flocks other

than those 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) would be 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) would be amended as follows:

a. In paragraph (b)(3)(i), the words ``, except turkey

hatcheries,'' would be removed.

b. In paragraph (b)(3)(ii) the words ``, except turkey flocks,''

would be removed.

c. In paragraph (b)(3)(viii), the words ``, other than turkey

flocks,'' would be removed.

d. In paragraph (b)(4), the words ``, other than turkey flocks,''

would be removed.

19. The subpart heading for subpart F would be 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 would be 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 would be amended by

adding the words ``emus, rheas, and cassowaries,'' immediately after

the word ``ostriches,'' and a new paragraph (c) would be 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) would be 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 would continue 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 would be removed and reserved.

25. In Sec. 147.6, paragraph (a)(14) would be 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 would be redesignated as footnotes 7

through 23, respectively.

27. A new Sec. 147.9 would be 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 immunodifussion Bull. Wld.

Hlth. Orig. 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

(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.

[[Page 43311]]

(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).

(x) Pipetting device capable of delivering 50 l 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

(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 (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 would be 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.'' would be added after the

final sentence.

b. In paragraph (b)(2)(v), the words ``at the National Veterinary

Services Laboratory'' would be removed.

29. A new Sec. 147.18 would be 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 35 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) would be

redesignated as paragraphs (d)(3) through (d)(6), respectively, and new

paragraphs (d)(1), (d)(2), (d)(7), and (d)(8) would be 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 National Poultry

Improvement Plan 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 would be amended by removing the words ``and E''

and adding the words ``E, and F'' in their place.

32. In Sec. 145.46, the introductory text of paragraph (a) would be

amended by removing the word ``four'' and adding the word ``five'' in

its place, and a new paragraph (a)(5) would be 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 4th day of August 1999.

Bobby R. Acord,

Acting Administrator, Animal and Plant Health Inspection Service.

[FR Doc. 99-20540 Filed 8-9-99; 8:45 am]

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