[Federal Register: September 16, 2008 (Volume 73, Number 180)]
[Page 53406-53407]
From the Federal Register Online via GPO Access [wais.access.gpo.gov]

                                                Federal Register

This section of the FEDERAL REGISTER contains documents other than rules 
or proposed rules that are applicable to the public. Notices of hearings 
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delegations of authority, filing of petitions and applications and agency 
statements of organization and functions are examples of documents 
appearing in this section.


[[Page 53406]]


Food Safety and Inspection Service

[Docket No. FSIS-2008-0007]

Revised FSIS Testing Methodology for Escherichia coli (E. coli) 

AGENCY: Food Safety and Inspection Service, USDA.

ACTION: Notice.


SUMMARY: The Food Safety and Inspection Service (FSIS) is announcing 
that, since January 28, 2008, its laboratories have implemented a 
revised laboratory methodology for detecting and identifying E. coli 
O157:H7 in regulatory verification samples. This new method can be 
found in the Microbiology Laboratory Guidebook (MLG), Chapter 5.04.

DATES: Submit comments by November 17, 2008.

ADDRESSES: FSIS invites interested persons to submit comments on this 
notice. Comments may be submitted by any of the following methods:
     Federal eRulemaking Portal: This Web site provides the 
ability to type short comments directly into the comment field on this 
Web page or attach a file for lengthier comments. Go to http://
www.regulations.gov and, in the ``Search Documents'' box type the 
docket number, FSIS-2008-0007. After accessing the docket, click on 
``Send a Comment or Submission.''
     Mail, including floppy disks or CD-ROMs, and hand- or 
courier-delivered items: Send to Docket Clerk, U.S. Department of 
Agriculture (USDA), FSIS, 1400 Independence Avenue, SW., Room 2534 
South Building, Washington, DC 20250.
    All submissions received must include the Agency name and docket 
number FSIS-2008-0007.
    All comments submitted in response to this notice will be posted to 
Agency's Web site at http://www.fsis.usda.gov/regulations_&_policies/
2008_Notices_Index/index.asp. Individuals who do not wish FSIS to 
post their personal contact information--mailing address, e-mail 
address, telephone number--on the Internet may leave this information 
off of their comments. Comments will also be available for public 
inspection in the FSIS Docket Room at the address listed above between 
8:30 a.m. and 4:30 p.m., Monday through Friday.

FOR FURTHER INFORMATION CONTACT: For further information contact 
William K. Shaw, Jr., Ph.D., Office of Policy and Program Development, 
FSIS, USDA, Room 3533, South Building, 14th and Independence, SW., 
Washington, DC 20250-3700; Telephone (202) 205-0695, Fax (202) 720-
7027, E-mail: william.shaw@fsis.usda.gov.



    FSIS' revised laboratory methodology for detecting and identifying 
E. coli O157:H7 in regulatory verification samples can be found in the 
Microbiology Laboratory Guidebook (MLG), Chapter 5.04 protocol, now 
available on the FSIS Web site at the following URL: http://
The revised MLG 5.04 describes the use of an improved enrichment broth 
and confirmatory identification procedures for E. coli O157:H7. The 
revisions to the protocol include an updated limit of the level at 
which E. coli O157:H7 can be detected for various products, and new 
instructions for preparing sub-samples of beef manufacturing trimmings 
and other ground beef components, and for preparing environmental 
sponge samples for E. coli O157:H7 analysis.
    In recent months, FSIS, in collaboration with the USDA Agricultural 
Research Service, has conducted an extensive validation of an 
enrichment broth, mTSB+n (also known as Modified Tryptone Soya Broth 
with Novobiocin and casamino acids or Modified Trypticase Soy with 
Novobiocin and casamino acids) to replace the broth that it had been 
using for detection of E. coli O157:H7. This broth has been found to 
support more rapid growth of E. coli O157:H7 than the mEC+n broth that 
FSIS laboratories have been using in past years. Moreover, the mTSB+n 
broth is cost-effective and is used by many laboratories around the 
world for detection of E. coli O157:H7.
    The use of mTSB+n will allow FSIS laboratories greater flexibility 
in the conducting of screening test procedures because it allows for 
greater flexibility in incubation time than does the broth FSIS 
previously used. Also, for some samples, its use will allow screening 
tests to be conducted earlier on the second day of analysis because the 
incubation period using this broth can be less than the broth FSIS 
previously used. The ability to conduct screening test procedures 
earlier can assist in more efficient laboratory staffing. Use of the 
replacement enrichment broth will also allow FSIS to obtain results 
faster for some samples. In addition, its use will reduce the number of 
samples that need to be discarded when there are shipping problems that 
delay the arrival of samples.
    Based on testing with inoculated samples, FSIS believes the new 
enrichment procedure is likely to provide increased sensitivity for 
detection in some samples. Therefore, there may be some increase in the 
number of E. coli O157:H7-positive samples found by FSIS laboratories.
    FSIS is also implementing new technologies to make its confirmatory 
identification testing procedures more robust and comprehensive for 
accurately identifying E. coli O157:H7, including variant strains that 
react in an atypical manner. Because serological testing procedures may 
not provide reliable identification for some strains of E. coli 
O157:H7, the revised protocol in the MLG 5.04 includes the use in FSIS 
laboratories of genetic testing to determine ``O157'' and ``H7'' 
identity in circumstances where atypical serology or other reactions 
are encountered. FSIS will use genetic testing to determine if the E. 
coli isolate is positive for Shiga toxin production, positive for the 
Shiga toxin gene, or genetically determined to be ``H7''.
    Since January 28, 2008, FSIS laboratories have been using the 
following terminology and criteria in the MLG 5.04 in reporting E. coli 
O157:H7 sample results:
     Potential Positive--a sample that causes a positive 
reaction with the screen test.
     Presumptive Positive--a sample that has typical colonies, 
observed on

[[Page 53407]]

Rainbow Agar, and reacts specifically with O157 antiserum.
     Confirmed Positive--a biochemically-identified Escherichia 
coli isolate that is serologically or genetically determined to be 
``O157'' that meets at least one of the following criteria:

(1) Positive for Shiga toxin (ST) production
(2) Positive for the Shiga toxin gene (stx)
(3) Genetically determined to be ``H7''

    If an FSIS laboratory's confirmatory test results are insufficient 
to allow identification of E. coli O157:H7, then the isolate is 
transferred to the Outbreaks Section of the Eastern Laboratory 
Microbiology Branch (OSEL), or to an FSIS reference laboratory, for 
further testing prior to issuance of a report of the test results.

State Programs and Foreign Government Programs

    States that have their own meat inspection programs for meat 
products produced and transported solely within the State are required 
to have mandatory ante mortem and post mortem inspection, reinspection, 
and sanitation requirements that are at least equal to those in the 
Federal Meat Inspection Act (21 U.S.C. 661(a)(1)). Therefore, these 
States' sampling procedures and testing methods for E. coli O157:H7 in 
raw beef products must be at least as sensitive as FSIS' procedures and 
testing methods for E. coli O157:H7.
    Foreign countries that are eligible to export meat products to the 
United States must apply inspection, sanitary, and other standards that 
are equivalent to those that FSIS applies to those products (21 U.S.C. 
620). Thus, in evaluating a foreign country's meat inspection system to 
determine the country's eligibility to export products to the United 
States, FSIS considers whether the testing methods and procedures for 
E. coli O157:H7 that the country applies are equivalent to those that 
FSIS uses.

Establishment Testing or Testing on Behalf of Establishments (Non-FSIS 

    As discussed above, FSIS will use genetic testing to determine 
``O157'' and ``H7'' identity in circumstances where atypical serology 
or other reactions are encountered. The genetic testing now used by 
FSIS laboratories for identification of certain strains of E. coli 
O157:H7 detected in FSIS laboratories may pose some difficulties for 
non-FSIS laboratories. Some non-FSIS laboratories may not be able to 
conduct genetic analyses because they do not have the necessary 
resources. Moreover, the improper application of a less robust or 
inadequately validated genetic test by a laboratory might lead to a 
false negative or other misleading results.
    FSIS believes that alternative criteria for confirmatory 
identification of E. coli O157:H7 or E. coli O157:H7-like bacteria may 
be appropriate if it is equivalent to the criteria used by FSIS. To be 
regarded as equivalent to the FSIS criteria, a test would have to 
detect all hypothetical E. coli O157:H7 strains, typical or variant, 
that would be identified using FSIS' confirmatory testing procedures 
and criteria.
    For establishment testing or testing on behalf of an establishment, 
FSIS recognizes that other criteria, while not used specifically by 
FSIS for identification of E. coli O157:H7, may be a significant and 
expedient indicator of the presence of E. coli O157:H7 in products. 
Such tests might be applied as rapid screening procedures to expedite 
analyses. If an establishment uses or contracts with a laboratory that 
uses such rapid screening procedures, and product is found positive by 
that test, FSIS expects the establishment to take appropriate 
corrective action and to ensure the proper disposition of adulterated 
products following a positive test result (9 CFR 417.3).

Additional Public Notification

    Public awareness of all segments of rulemaking and policy 
development is important. Consequently, in an effort to ensure that 
minorities, women, and persons with disabilities are aware of this 
notice, FSIS will announce it online through the FSIS Web page located 
at http://www.fsis.usda.gov/regulations/2008_Notices_Index/. FSIS 
will also make copies of this Federal Register publication available 
through the FSIS Constituent Update, which is used to provide 
information regarding FSIS' policies, procedures, regulations, Federal 
Register notices, public meetings, and other types of information that 
could affect or would be of interest to constituents and stakeholders. 
The Update is communicated via Listserv, a free electronic mail 
subscription service for industry, trade groups, consumer interest 
groups, health professionals, and other individuals who have asked to 
be included. The Update is also available on the FSIS Web page. Through 
the Listserv and Web page, FSIS is able to provide information to a 
much broader and more diverse audience. In addition, FSIS offers an e-
mail subscription service which provides automatic and customized 
access to selected food safety news and information. This service is 
available at http://www.fsis.usda.gov/news_and_events/email_
subscription/. Information is available about a variety of topics 
including recalls, exports, regulations, directives, and notices. 
Customers can add or delete subscriptions themselves, and they have the 
option to password protect their accounts.

    Done at Washington, DC, on September 4, 2008.
Alfred V. Almanza,
[FR Doc. E8-21575 Filed 9-15-08; 8:45 am]