[Federal Register Volume 75, Number 234 (Tuesday, December 7, 2010)]
[Notices]
[Pages 76008-76009]
[FR Doc No: 2010-30582]

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DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

[Docket No. FDA-2010-N-0606]

Agency Information Collection Activities; Proposed Collection;
Comment Request; Additional Listing Information for Medical Device
Registration and Listing

AGENCY: Food and Drug Administration, HHS.

ACTION: Notice.

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SUMMARY: The Food and Drug Administration (FDA) is announcing an
opportunity for public comment on the proposed collection of certain
information by the Agency. Under the Paperwork Reduction Act of 1995
(the PRA), Federal Agencies are required to publish notice in the
Federal Register concerning each proposed collection of information,
including each proposed extension of an existing collection of
information, and to allow 60 days for public comment in response to the
notice. This notice solicits comments on the reporting and
recordkeeping burden requirements associated with additional listing
information for medical device registration and listing by non-
electronic means.

DATES: Submit either electronic or written comments on the collection
of information by February 7, 2011.

ADDRESSES: Submit electronic comments on the collection of information
to http://www.regulations.gov. Submit written comments on the
collection of information to the Division of Dockets Management (HFA-
305), Food and Drug Administration, 5630 Fishers Lane, rm. 1061,
Rockville, MD 20852. All comments should be identified with the docket
number found in brackets in the heading of this document.

FOR FURTHER INFORMATION CONTACT: Daniel Gittleson, Office of
Information Management Food and Drug Administration, 1350 Piccard Dr.,
PI50-400B, Rockville, MD 20850,<PHONE_NUMBER>,
<EMAIL_ADDRESS>
SUPPLEMENTARY INFORMATION: Under the PRA (44 U.S.C. 3501-3520), Federal
Agencies must obtain approval from the Office of Management and Budget
(OMB) for each collection of information they conduct or sponsor.
``Collection of information'' is defined in 44 U.S.C. 3502(3) and 5 CFR
1320.3(c) and includes Agency requests or requirements that members of
the public submit reports, keep records, or provide information to a
third party. Section 3506(c)(2)(A) of the PRA (44 U.S.C. 3506(c)(2)(A))
requires Federal Agencies to provide a 60-day notice in the Federal
Register concerning each proposed collection of information, including
each proposed extension of an existing collection of information,
before submitting the collection to OMB for approval. To comply with
this requirement, FDA is publishing notice of the proposed collection
of information set forth in this document.
    With respect to the following collection of information, FDA
invites comments on these topics: (1) Whether the proposed collection
of information is necessary for the proper performance of FDA's
functions, including whether the information will have practical
utility; (2) the accuracy of FDA's estimate of the burden of the
proposed collection of information, including the validity of the
methodology and assumptions used; (3) ways to enhance the quality,
utility, and clarity of the information to be collected; and (4) ways
to minimize the burden of the collection of information on respondents,
including through the use of automated collection techniques, when
appropriate, and other forms of information technology.

Additional Listing Information for Medical Device Registration and
Listing (OMB Control Number 0910-0387)--Extension

    The Food and Drug Administration Amendments Act of 2007 (FDAAA),
enacted September 27, 2007, requires that device establishment
registrations and listings under 21 U.S.C. 360(p) (including the
submission of updated information) be submitted to the Secretary of
Health and Human Services (the Secretary) by electronic means,

unless the Secretary grants a request for waiver of the requirement,
because the use of electronic means is not reasonable for the person
requesting the waiver. The collections of information under sections
222, 223, and 224 of FDAAA have been approved under OMB control number
0910-0625. Registration by electronic means for device establishments
replaced FDA Forms 2891 and 2891a, ``Registration of Device
Establishment,'' and FDA Form 2892, ``Medical Device Listing,'' with
FDA Form 3673, ``Device Registration and Listing Module.'' The scope of
this information collection addresses only the reporting and
recordkeeping requirements by non-electronic means under Sec.  807.31
(21 CFR 807.31).
    Under Sec.  807.31(a) through (d), each owner or operator is
required to maintain an historical file containing the labeling and
advertisements in use on the date of initial listing, and in use after
October 10, 1978, but not before the date of initial listing. The owner
or operator must maintain in the historical file any labeling or
advertisements in which a material change has been made anytime after
initial listing, but may discard labeling and advertisements from the
file 3 years after the date of the last shipment of a discontinued
device by an owner or operator. Section 807.31(e) requires that the
owner or operator be prepared to submit to FDA copies of: (1) All
device labeling, (2) all device labeling and representative
advertising, or (3) only representative package inserts, depending upon
whether the device is subject to the regulatory controls under sections
514 and 515 of the Federal Food, Drug, and Cosmetic Act (the FD&C Act)
(21 U.S.C. 360d and 360e), or restrictions imposed by 21 CFR 801.109 or
otherwise by section 520(e) of the FD&C Act.
    The information collected under these provisions is used by FDA to
identify: (1) Firms subject to FDA's regulations, (2) geographic
distribution of firms in order to effectively allocate FDA's field
resources for inspections, and (3) the class of the device that
determines the frequency of inspection. As a result, when complications
occur with a particular device or component, all manufacturers of
similar or related devices can easily be identified.
    The likely respondents to this information collection are domestic
and foreign device establishments who must register and submit a device
list to FDA, e.g., establishments engaged in the manufacture,
preparation, propagation, compounding, assembly, or processing of
medical devices intended for human use and commercial distribution.
    FDA estimates the burden of this collection of information as
follows:

                                 Table 1--Estimated Annual Reporting Burden \1\
----------------------------------------------------------------------------------------------------------------
                                                      Annual
         21 CFR Section              Number of     frequency of    Total annual      Hours per      Total hours
                                    respondents      response        responses       response
----------------------------------------------------------------------------------------------------------------
807.31(d)(2)....................           2,250               1           2,250              .5           1,125
807.31(e).......................          22,500               1          22,500              .5          11,250
                                 -------------------------------------------------------------------------------
    Total.......................  ..............  ..............  ..............  ..............          12,375
----------------------------------------------------------------------------------------------------------------
\1\ There are no capital costs or operating and maintenance costs associated with this collection of
  information.

                               Table 2--Estimated Annual Recordkeeping Burden \1\
----------------------------------------------------------------------------------------------------------------
                                                      Annual
         21 CFR Section              Number of     frequency per   Total annual      Hours per      Total hours
                                   recordkeepers   recordkeeping      records         record
----------------------------------------------------------------------------------------------------------------
807.31(a-c).....................          22,500               4          90,000            0.50          45,000
                                 -------------------------------------------------------------------------------
    Total.......................  ..............  ..............  ..............  ..............          45,000
----------------------------------------------------------------------------------------------------------------
\1\ There are no capital costs or operating and maintenance costs associated with this collection of
  information.

    The annual respondent reporting burden for device establishment
registrations and listings for additional information is estimated to
be 12,375 hours and the annual respondent recordkeeping burden is
estimated to be 45,000 hours. Therefore, the total burden hours for
this collection are estimated to be 57,375. The estimates cited in
tables 1 and 2 of this document are based primarily on fiscal year 2010
data from current systems and on conversations with industry and trade
association representatives.

    Dated: December 1, 2010.
Leslie Kux,
Acting Assistant Commissioner for Policy.
[FR Doc. 2010-30582 Filed 12-6-10; 8:45 am]
BILLING CODE 4160-01-P


[Federal Register: September 12, 2008 (Volume 73, Number 178)]
[Notices]
[Page 53062-53064]
[DOCID:fr12se08-110]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-58462; File No. SR-CBOE-2008-91]

Self-Regulatory Organizations; Chicago Board Options Exchange,
Inc.; Notice of Filing and Immediate Effectiveness of Proposed Rule
Change by Chicago Board Options Exchange, Inc. Amending CBOE Rule 52.3

September 4, 2008.
    Pursuant to Section 19(b)(1) \1\ of the Securities Exchange Act of
1934 (the ``Act'') \2\ and Rule 19b-4 thereunder,\3\ notice is hereby
given that on August 27, 2008, Chicago Board Options Exchange, Inc.
(``CBOE'' or the ``Exchange'') filed with the Securities and Exchange
Commission (``Commission'') the proposed rule change described in Items
I, and II below, which Items have been prepared by the Exchange. The
Commission is publishing this notice to solicit comments on the
proposed rule change from interested persons.
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    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 15 U.S.C. 78a.
    \3\ 17 CFR 240.19b-4.
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I. Self-Regulatory Organization's Statement of the Terms of Substance
of the Proposed Rule Change

    The Exchange proposes to amend CBOE Rule 52.3 in order to clarify a
circumstance under which the Exchange will commence a trading halt in
Derivative Securities Products listed and trading on the Exchange.

II. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

    In its filing with the Commission, the self-regulatory organization
included statements concerning the purpose of, and basis for, the
proposed rule change and discussed any comments it received on the
proposed rule change. The text of these statements may be examined at
the places specified in Item IV below. The Exchange has prepared
summaries, set forth in sections A, B, and C below, of the most
significant parts of such statements.

A. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

1. Purpose
    In order to provide clarity to CBSX Traders,\4\ the Exchange
proposes to amend its Rules by adding new CBOE Rule 52.3(d), in order
to clarify that the Exchange will halt trading in Derivative Securities
Products \5\ listed and trading on the CBOE Stock Exchange \6\
(``CBSX'') when the Exchange becomes aware that the Net Asset Value
(``NAV'') of the Derivative Securities Products (and in the case of
managed fund shares, the disclosed portfolio) is not being disseminated
to all market participants at the same time.
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    \4\ See CBOE Rule 50.3 for the definition of ``CBSX Traders.''
    \5\ Pursuant to Rule 52.3(c)(5)(i), the term Derivative
Securities Product includes a series of Equity-Linked Term Notes,
Index-Linked Exchangeable Notes, IPRs, IPSs, or TIRs, which is based
on an underlying security or index. Those products are described in
CBOE Rules 31.5I, 31.5O, 31.5L, 31.5M, and 31.5N, respectively.
    \6\ The CBOE Stock Exchange is the stock trading facility of
CBOE.
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    Specifically, with respect to Derivative Securities Products listed
on CBSX for which an NAV (and in the case of managed fund shares, a
disclosed portfolio) is disseminated, if the Exchange becomes aware
that the NAV (or in the case of managed fund shares, the disclosed
portfolio) is not being disseminated to all market participants at the
same time, it will halt trading in the affected Derivative Securities
Product on CBSX until such time as the NAV (or in the case of managed
fund shares, the disclosed portfolio, as applicable) is available to
all market participants. The Exchange believes that adding this
clarification to its rules promotes the just and equitable principles
of trade and promotes a fair and transparent marketplace.
    The Exchange notes that, in the event the NAV (or disclosed
portfolio, as applicable) for one of the securities referred to above
is no longer calculated or disseminated, the Exchange would halt
trading in such security and would resume trading at such time as the
NAV (or disclosed portfolio, as applicable) is available.
2. Statutory Basis
    The Exchange believes that the proposed rule change is consistent
with and furthers the objectives of Section 6(b)(5) of the Act,\7\ in
that it is designed to prevent fraudulent and manipulative acts and
practices, to promote just and equitable principles of trade, to remove
impediments to, and perfect the mechanism of, a free and open market
and a national market system, and, in general, to protect investors and
the public interest.
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    \7\ 15 U.S.C. 78f(b)(5).
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B. Self-Regulatory Organization's Statement on Burden on Competition

    The Exchange does not believe that the proposed rule change will
impose any burden on competition that is not necessary or appropriate
in furtherance of the purpose of the Act.

C. Self-Regulatory Organization's Statement on Comments on the Proposed
Rule Change Received From Members, Participants or Others

    No written comments were solicited or received with respect to the
proposed rule change.

III. Date of Effectiveness of the Proposed Rule Change and Timing for
Commission Action

    The Exchange has designated the proposed rule change as one that:
(i) Does not significantly affect the protection of investors or the
public interest; (ii) does not impose any significant burden on
competition; and (iii) by its terms, does not become operative for 30
days from the date on which it was filed, or such shorter time as the
Commission may designate if consistent with the protection of investors
and the public interest. Additionally, the Exchange provided the
Commission with written notice of its intention to file the proposed
rule change at least five business days before its filing. Therefore,
the foregoing rule change has become effective pursuant to Section
19(b)(3)(A) of the Act \8\ and Rule 19b-4(f)(6) thereunder.\9\
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    \8\ 15 U.S.C. 78s(b)(3)(A).
    \9\ 17 CFR 240.19b-4(f)(6).
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    The Exchange has requested that the Commission waive the 30-day
operative delay, which would make the rule change operative upon
filing. The Exchange believes that waiver of the 30-day operative delay
is consistent with the protection of investors and the public interest
as it will clarify a circumstance under which the Exchange will halt
trading in Derivative Securities Products listed on the Exchange. This
proposal is similar to rule changes by other national securities
exchanges, and does not raise any new or unique issues.\10\ Therefore,
the Commission designates the proposed rule change as operative
immediately.\11\
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    \10\ See Securities Exchange Act Release No. 58111 (July 7,
2008), 73 FR 40643 (July 15, 2008) (SR-Amex-2008-40; SR-NASDAQ-2008-
046; SR-NYSE-2008-39; SR-NYSEArca-2008-50).
    \11\ For purposes only of waiving the operative delay for this
proposal, the Commission has considered the proposed rule's impact
on efficiency, competition, and capital formation. See 15 U.S.C.
78c(f).
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IV. Solicitation of Comments

    Interested persons are invited to submit written data, views, and
arguments concerning the foregoing, including whether the proposed rule
change is consistent with the Act. Comments may be submitted by any of
the following methods:

Electronic Comments

     Use the Commission's Internet comment form (http://
www.sec.gov/rules/sro.shtml); or
     Send an e-mail to<EMAIL_ADDRESS>Please include
File Number SR-CBOE-2008-91 on the subject line.

Paper Comments

     Send paper comments in triplicate to Secretary, Securities
and Exchange Commission, 100 F Street, NE., Washington, DC 20549-1090.

All submissions should refer to File Number SR-CBOE-2008-91. This file
number should be included on the subject line if e-mail is used. To
help the Commission process and review your comments more efficiently,
please use only one method. The Commission will post all comments on
the Commission's Internet Web site (http://www.sec.gov/rules/
sro.shtml). Copies of the submission, all subsequent amendments, all
written statements with respect to the proposed rule change that are
filed with the Commission, and all written communications relating to
the proposed rule change between the Commission and any person, other
than those that may be withheld from the public in accordance with the
provisions of 5 U.S.C. 552, will be available for inspection and
copying in the Commission's Public Reference Room, 100 F Street, NE.,
Washington, DC 20549, on official business days between the hours of 10
a.m. and 3 p.m. Copies of the filing also will be available for
inspection and copying at the principal office of the self-regulatory
organization. All comments received will be posted without change; the
Commission does not edit personal identifying information from
submissions. You should submit only information that you wish to make
available publicly. All submissions should refer to File Number SR-
CBOE-

2008-91 and should be submitted on or before October 3, 2008.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\12\
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    \12\ 17 CFR 200.30-3(a)(12).
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Florence E. Harmon,
Acting Secretary.
[FR Doc. E8-21167 Filed 9-11-08; 8:45 am]

BILLING CODE 8010-01-P


Memo to Docket  Subject: Call Between Freeport McMoRan Miami Inc. (FMMI) and EPA Date: May 28, 2014

Participants: Name:

Organization:

Chris West

Freeport-McMoRan

Jeff Harrington

AMEC (FMMI contractor)

Margaret Peloso

Vinson & Elkins (external counsel)

Todd Weaver

Freeport-McMoRan

Eugene Chen

EPA R9  Geoffrey Glass

EPA R9 Charlotte Withey Colleen McKaughan

EPA R9 EPA R9

Summary: 	    * This call was requested by FMMI to discuss the compliance deadline for the Miami Smelter in the Arizona RH FIP.    * FMMI described need for shakedown period and reiterated request for a compliance date four years from publication of final rule.    * FMMI described possible interim requirements.


[Federal Register: October 22, 2010 (Volume 75, Number 204)]
[Notices]
[Page 65312]
[DOCID:fr22oc10-58]

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DEPARTMENT OF ENERGY

Federal Energy Regulatory Commission

[Project No. 13831-000]

Vortex Hydro Energy, LLC; Notice of Competing Preliminary Permit
Application Accepted for Filing and Soliciting Comments and
Interventions

October 15, 2010.
    On August 9, 2010, Vortex Hydro Energy, LLC filed an application
for a preliminary permit, pursuant to section 4(f) of the Federal Power
Act, proposing to study the feasibility of the St. Clair Hydrokinetic
Project, located on the St. Clair River, in St. Clair County, Michigan.
The sole purpose of a preliminary permit, if issued, is to grant the
permit holder priority to file a license application during the permit
term. A preliminary permit does not authorize the permit holder to
perform any land disturbing activities or otherwise enter upon lands or
waters owned by others without the owners' express permission.
    The proposed project would consist of: (1) Two medium scale VIVACE
converters with each unit consisting of approximately nine cylinders
that is gravity held at the bottom of the river bed; (2) an underwater
inverter that would convert the electric signal to 3 Phase 480 Volts;
and (3) a proposed underwater cable that would transmit to the nearby
Dunn Paper Plant. The proposed St. Clair Hydrokinetic Project would
have an estimated average annual generation of 0.788 gigawatt-hours.
    Applicant Contact: Gus Simiao, CEO, Vortex Hydro Energy, LLC, 2512
Carpenter Road, Suite 201-A1, Ann Arbor, MI 48108; phone: (734) 971-
4020.
    FERC Contact: Bryan Roden-Reynolds at<PHONE_NUMBER>, or via e-mail
at<EMAIL_ADDRESS>    Competing Applications: This application competes with Project No.
13694-000 filed March 30, 2010.
    Deadline for filing comments and motions to intervene: 60 days from
the issuance of this notice. Comments and motions to intervene may be
filed electronically via the Internet. See 18 CFR 385.2001(a)(1)(iii)
and the instructions on the Commission's Web site (http://www.ferc.gov/
docs-filing/ferconline.asp) under the ``eFiling'' link. For a simpler
method of submitting text only comments, click on ``eComment.'' For
assistance, please contact FERC Online Support at
<EMAIL_ADDRESS>call toll free at<PHONE_NUMBER>; or, for
TTY, contact<PHONE_NUMBER>. Although the Commission strongly
encourages electronic filing, documents may also be paper-filed. To
paper-file, mail an original and eight copies to: Kimberly D. Bose,
Secretary, Federal Energy Regulatory Commission, 888 First Street, NE.,
Washington, DC 20426.
    More information about this project, including a copy of the
application, can be viewed or printed on the ``eLibrary'' link of the
Commission's Web site at http://www.ferc.gov/docs-filing/elibrary.asp.
Enter the docket number (P-13617) in the docket number field to access
the document. For assistance, contact FERC Online Support.

Kimberly D. Bose,
Secretary.
[FR Doc. 2010-26690 Filed 10-21-10; 8:45 am]
BILLING CODE 6717-01-P


[Federal Register Volume 85, Number 118 (Thursday, June 18, 2020)]
[Notices]
[Pages 36910-36911]
[FR Doc No: 2020-13117]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-89057; File No. SR-NYSEArca-2019-77]

Self-Regulatory Organizations; NYSE Arca, Inc.; Notice of
Designation of a Longer Period for Commission Action on Proceedings To
Determine Whether To Approve or Disapprove a Proposed Rule Change To
List and Trade Shares of the AdvisorShares Pure U.S. Cannabis ETF Under
NYSE Arca Rule 8.600-E

June 12, 2020.
    On December 13, 2019, NYSE Arca, Inc. (``NYSE Arca'') filed with
the Securities and Exchange Commission (``Commission''), pursuant to
Section 19(b)(1) of the Securities Exchange Act of 1934 (``Act'') \1\
and Rule 19b-4 thereunder,\2\ a proposed rule change to list and trade
shares of the AdvisorShares Pure U.S. Cannabis ETF under NYSE Arca Rule
8.600-E (Managed Fund Shares). The proposed rule change was published
for comment in the Federal Register on December 26, 2019.\3\ On January
28, 2020, pursuant to Section 19(b)(2) of the Act,\4\ the Commission
designated a longer period within which to approve the proposed rule
change, disapprove the proposed rule change, or institute proceedings
to determine whether to approve or disapprove the proposed rule
change.\5\ On March 13, 2020, the Commission

instituted proceedings pursuant to Section 19(b)(2)(B) of the Act \6\
to determine whether to approve or disapprove the proposed rule
change.\7\ The Commission has received no comment letters on the
proposed rule change.
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    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 17 CFR 240.19b-4.
    \3\ See Securities Exchange Act Release No. 87791 (December 18,
2019), 84 FR 71057.
    \4\ 15 U.S.C. 78s(b)(2).
    \5\ See Securities Exchange Act Release No. 88066, 85 FR 6009
(February 3, 2020).
    \6\ 15 U.S.C. 78s(b)(2)(B).
    \7\ See Securities Exchange Act Release No. 88378, 85 FR 15834
(March 19, 2020).
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    Section 19(b)(2) of the Act \8\ provides that, after initiating
disapproval proceedings, the Commission shall issue an order approving
or disapproving the proposed rule change not later than 180 days after
the date of publication of notice of filing of the proposed rule
change. The Commission may extend the period for issuing an order
approving or disapproving the proposed rule change, however, by not
more than 60 days if the Commission determines that a longer period is
appropriate and publishes the reasons for such determination. The
proposed rule change was published for notice and comment in the
Federal Register on December 26, 2019. June 23, 2020 is 180 days from
that date, and August 22, 2020 is 240 days from that date.
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    \8\ 15 U.S.C. 78s(b)(2).
---------------------------------------------------------------------------

    The Commission finds it appropriate to designate a longer period
within which to issue an order approving or disapproving the proposed
rule change so that it has sufficient time to consider the proposed
rule change. Accordingly, the Commission, pursuant to Section 19(b)(2)
of the Act,\9\ designates August 22, 2020 as the date by which the
Commission shall either approve or disapprove the proposed rule change
(File No. SR-NYSEArca-2019-77).
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    \9\ Id.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\10\
---------------------------------------------------------------------------

    \10\ 17 CFR 200.30-3(a)(57).
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J. Matthew DeLesDernier,
Assistant Secretary.
[FR Doc. 2020-13117 Filed 6-17-20; 8:45 am]
BILLING CODE 8011-01-P


Interim Quantitative Method for Evaluating the Efficacy of Antimicrobial Test Substances on Porous Surfaces Against Bacteria
                                 (12/08/2022)

Scope
The Environmental Protection Agency (EPA) Office of Pesticide Programs (OPP) recommends that applicants utilize this interim method to support efficacy criteria for the registration of products bearing claims for use on soft, porous surface claims.  The method provides a quantitative assessment of the performance of antimicrobial substances against Pseudomonas aeruginosa and Staphylococcus aureus on soft-porous surfaces.
This method provides log reduction (LR) as the quantitative measure of efficacy for disinfectants against the test microbes on a soft-porous surface.

Method Overview
In brief, the method uses 1 cm diameter discs (carriers) of a set of representative soft-porous surface materials.  Each disc receives 10 uL of microbial inoculum (with a three-part organic and inorganic soil load) deposited in the center of each carrier. The inoculum is allowed to dry and is then exposed to 50 uL of the antimicrobial treatment; control carriers receive an equivalent volume of an innocuous fluid (e.g., phosphate buffered saline). The exposure time is allowed to elapse; a liquid neutralizer is then added to the vial to halt the antimicrobial action. Each vial with the carrier is vortexed, serially diluted, and the contents are filtered to recover viable microorganisms. Based on the difference between the mean log10 density values of the untreated control and treated carriers, a mean log10 reduction (LR) in viable bacteria is calculated. The LR value is used as the measure of product effectiveness.
      Appropriate safety procedures should always be used when working with laboratory test systems which include human pathogenic microorganisms. Laboratory safety is discussed in the current edition of "Biosafety in Microbiological and Biomedical Laboratories (BMBL)" 6[th] edition, from the subject matter experts within the U.S. Department of Health and Human Services (HHS), including experts from the Centers for Disease Control and Prevention (CDC) and National Institutes of Health (NIH).
                               TABLE OF CONTENTS

1) Special Apparatus and Materials...............................................................
3
2) Carriers.............................................................................................
6
3) Preparation of Test Cultures and Carrier Inoculation ......................................
7
4) Performance Assessment  -  Efficacy..........................................................
9
5) Data Requirements...............................................................................
12
Attachment A: Preparation of Frozen Stock Culture...........................................
14
Attachment B: Neutralization Assay and Flowcharts...........................................
16

 Special Apparatus and Materials
 Test microbes: Pseudomonas aeruginosa (ATCC #15442) and Staphylococcus aureus (ATCC #6538).
 Additional bacteria may be tested (for an additional label claim) per the Agency's guidance.
 Culture media
 Tryptic Soy Broth (TSB).  Use to rehydrate lyophilized cultures. Purchase broth from a reputable source or prepare according to manufacturer's instructions.
 Synthetic broth (SB). Growth medium for test cultures.  Commercial media (HIMEDIA, Synthetic Broth, AOAC, #M334-500G). Suspend 16.9 g in 1000 mL DI water. Heat if necessary, to dissolve the medium completely. Final pH at 25°C should be 7.1+-02. Medium may be dispensed in 10 mL amounts in 20x150 mm culture tubes or alternatively in 500 mL volumes in a 1 L bottle; steam sterilize at 15 lbs pressure (121°C) for 15 minutes. Cool to room temperature and just before use, aseptically add 0.1 mL of 10% sterile dextrose solution. Store prepared SB at 2-8°C.
 Alternatively, SB made in-house per the recipe provided in AOAC Methods 955.15, 964.02, and 955.14 may be substituted.
 10% dextrose solution. Add 5.0 g dextrose to 50 mL de-ionized water and mix by stirring. Filter sterilize the solution using a 0.2 um filter. Store the sterile solution at 2-5°C for up to 30 days.
 TSB with 15% (v/v) glycerol. Use as a cryoprotectant.  Suspend 7.5 g tryptic soy broth in 212.5 mL de-ionized water. Add 37.5 mL glycerol and stir, warm slightly to dissolve. Dispense into bottles and steam sterilize for 15 min at 121°C.
 Tryptic soy agar (TSA) and TSA with 5% sheep blood.  Use for culturing, isolation, and characterization of the test microbes. Purchase plates from a reputable source or prepare according to manufacturer's instructions.
 Selective media. (optional) Mannitol salt agar and Cetrimide agar. Use for quality control of test microbes listed in this procedure. Purchase plates or prepare according to manufacturer's instructions.
 Reagents
 Neutralizer.  A liquid reagent used to inactivate and/or dilute the antimicrobial treatment to end the contact time.
 Phosphate buffered saline stock solution (e.g., 10X). To prepare 1X phosphate buffered saline. The stock solution has a pH of approximately 7.2+-0.2.
 Phosphate buffered saline (PBS), 1X. Dilution blanks and filtration. PBS with a pH of approximately 7.0+-0.5 is desirable.
 Soil load, 3-part. Use as the soiling agent. Add to the test suspension in the following manner:
 BSA: Add 0.5 g bovine serum albumin (BSA, radio immunoassay (RIA) grade or equivalent, CAS# 9048-46-8) to 10 mL of PBS, mix and pass through a 0.2 um pore diameter (polyethersulfone) membrane filter, aliquot (e.g., a minimum of 50 uL), and store at -20+-2°C.
 Yeast Extract: Add 0.5 g yeast extract to 10 mL of PBS, mix, and pass through a 0.2 um pore diameter (polyethersulfone) membrane filter, aliquot (e.g., a minimum of 70 uL), and store at -20+-2°C.
 Mucin: Add 0.04 g mucin (from bovine submaxillary gland, CAS # 84195-52-8) to 10 mL of PBS, stir or vortex-mix until thoroughly dissolved, and pass through a 0.2 um pore diameter (polyethersulfone) membrane filter, aliquot, and store at -20+-2°C.
 The three stock solutions of the soil load are single use only. Do not refreeze; store up to one year at -20+-2°C.
 Antimicrobial Test substance. Ready-to-use, activated, or concentrated antimicrobial. If the antimicrobial test substance is prepared by diluting a concentrate in hard water, then use prepared test substance within 3 hours of preparation or as otherwise instructed by the manufacturer. Measuring error increases as delivery volume decreases. To minimize variability due to measuring error, a minimum of 1.0 mL or 1.0 g of concentrated antimicrobial test substance should be used when preparing use-dilutions for testing. Use v/v dilutions for liquids antimicrobial test substances and w/v dilutions for solid antimicrobial test substances. The use of a positive displacement pipette is recommended for viscous liquids.
 1 N NaOH and 1 N HCl. Used for pH adjustment of media/reagents.
 375 ppm hard water. Used for the preparation of antimicrobial test substances.  Adjust the recipe for volumes other than 1 L.
 Prepare Solution A by dissolving 19.84 g anhydrous magnesium chloride (or 42.36 g MgCl·6H2O) and 46.24 g anhydrous calcium chloride (CaCl2) in de-ionized water and dilute to 1,000 mL. Sterilize by membrane filtration. Store the solution in the refrigerator (2-8°C) for up to 30 days; do not use after that time.
 Prepare Solution B by dissolving 35.02 g sodium bicarbonate (NaHCO3) in water and dilute to 1,000 mL. Sterilize by membrane filtration. Store the solution in the refrigerator (2-8°C) for up to 30 days; do not use after that time.
 To prepare 1 L of 375 ppm hard water, place 600-700 mL of deionized water in a 1,000 mL volumetric flask and add 6.0 mL of Solution A and then 8.0 mL of Solution B. Mix and add water to the flask to reach 1,000 mL. The pH of the hard water should be 7.0+-0.2 at room temperature. If necessary, adjust the pH by using 1 N NaOH or 1 N HCl.  Ensure sterility of hard water.
 Prepare the hard water under aseptic conditions and use within 5 days of preparation. On the day of the test, measure the hardness of the water using a water hardness test kit or other suitable titration method.
 The target hardness expressed as mg/L calcium carbonate (CaCO3) is 375 mg/L +5%/-10% (338-394 ppm).
 Water. De-ionized (DI), distilled water or water with equivalent quality for making reagent solutions and culture media.
 Tween-80 (polysorbate 80). Used to prepare PBS-T.
 Gram stain. Used for diagnostic staining.
 Apparatus
 Carriers: Discs (1 cm in diameter) cut from porous material.  Carriers are single use only.  See Section 2 for carrier specifications.
 Hole punch: If necessary, for use in the preparation of 1 cm disc from material.  Model number: SKU# HP-MEI448R or equivalent
 Calibrated 10 uL positive displacement pipette with corresponding 10 uL tips, for carrier inoculation.
 Filter paper. Whatman No. 2, to line glass Petri plates.
 Calibrated micropipettes (e.g., 200 uL, 1 mL) with appropriate corresponding tips, for deposition of test substance on carriers and preparing dilutions.
 Bottle-top dispensers, squirt bottles, pre-measured volumes in tubes, or pipettes, bottles, etc. For rinsing vials and filters.
 Forceps, straight or curved, non-magnetic, disposable with smooth flat tips to handle membrane filters, appropriate to pick up the carriers for placement in vials.
 Polyethersulfone (PES) membranes.  Use for recovery of test microbe, 47 mm diameter and 0.2 um pore size.
 Use filter membranes in either a reusable or disposable filtration unit.
 Filter Sterilization Unit (with PES, 0.2 um pore size).  Use to filter sterilize soil components.
 20 x 150 m glass culture tubes with Morton closures for test culture preparation.
 Spectrophotometer. For culture standardization (if deemed necessary)
 Vials with lids (plastic or comparable). Sterile, flat-bottomed, wide-mouthed (at least 25 mm diameter), approximately 20 mL capacity, for holding inoculated carriers to be exposed to the test chemical and for accommodating neutralizer.
 Transparent vials are more desirable to facilitate application of 50 uL test substance or control substance to inoculated carrier.
 Certified timer. Readable in minutes and seconds, for tracking of timed events and intervals.
 Desiccation unit (with gauge to measure vacuum level) with fresh desiccant (e.g., anhydrous CaCO3). For drying inoculated carriers.
 Vacuum source. In-house line or suitable vacuum pump capable of achieving 0.068 to 0.085 MPa, for drying inoculated carriers in desiccation unit and to perform membrane filtration.
 Titration kit. (i.e., Hach digital titrator) Used for measuring water hardness.
 Vortex-style mixer. Used for vortex-mixing of various solutions.
 15 mL conical centrifuge tubes. Used for centrifugation of test cultures.
 Centrifuge (with rotor capable of achieving 5,000g). Used for test culture preparation.
 Carriers
 Carrier Materials
 Privacy Curtain Fabric (PCF-03): 54% Polyester, 46% Fire Resistant (FR) Polyester. CF Stinson, LLC. Mambo MAM34 Nights.
 Non-PVC Fabric (NVF-01): Polyurethane Face made with Polycarbonate and Polyether Resins, Polyester Backing. CF Stinson, LLC. Kid BlueSky KID17.
 Vinyl Seating Fabric (VF-01): Vinyl Face, Polyester Backing. CF Stinson, LLC. Hopsack - HOP24 Fjord.
                                           Vinyl
                                    (VF-01)
                                Privacy Curtain
                                   (PCF-03)
                                    Non-PVC
                                   (NVF-01)
Figure 1: Examples of carrier materials cut into 1 cm discs; materials 2.a.i, 2.a.ii, and 2.a.iii (from left to right)
 Carrier Preparation
 Punch, or obtain, 1 cm round carriers or use comparable cutting procedure from fabric.
 Visually screen carriers to ensure consistent surface characteristics; trim any jagged edges or loose fabric.
 No pre-cleaning of carriers is necessary. To sterilize carriers, sterilize using a gravity cycle, 121°C for 20 minutes; ensure carriers are dry following sterilization.  Test sterility of carriers prior to testing.
 Carriers may not be entirely flat after autoclaving; however, minor distortion of carriers is acceptable for testing.
 Prior to use in testing, document the condition of the screened and sterile carriers (e.g., digital photographs).
 Preparation of Test Culture and Carrier Inoculation
 Refer to Attachment A for preparation of the frozen stock cultures.
 Defrost a cryovial; defrost rapidly to avoid loss in the viability of the preserved cells. Each cryovial is single use only.
 Within 15 minutes prior to inoculation, using a calibrated pipette to aseptically add 0.1 mL of 10% sterile dextrose (w/v) solution to each 10 mL tube of SB.
 Using a calibrated micropipette, add 100 uL of defrosted stock culture to 10 mL SB with dextrose, briefly vortex-mix and incubate for 24+-2 h at 36+-1°C.
 Incubate without disrupting the culture.
 In addition, inoculate an agar plate (e.g., TSA or TSA with 5% sheep blood) from the inoculated tube and streak for isolation. Incubate plate with the test culture.
 Following incubation, use the SB cultures to prepare a test suspension for each organism.
 The 24+-2 h culture should exhibit a titer of at least 10[8] CFU/mL.
 For P. aeruginosa, inspect culture prior to harvest; visible pellicle on the surface of the culture is expected to form during incubation (record its presence). Discard the culture if pellicle has been disrupted (fragments in culture).
 Remove visible pellicle on surface of medium and around associated interior edges of the tube by pipetting or with vacuum suction.
 Using a serological pipette, withdraw the remaining broth culture (at least 5 mL) avoiding any sediment on the bottom of the tube and transfer it into a 15 mL conical centrifuge tube.
 Record approximate volume harvested and transferred to 15 mL conical tube.
 For S. aureus, briefly vortex-mix the 24+-2 h culture and transfer to a 15 mL conical centrifuge tube.
 Within 15 min, centrifuge the 24+-2 h harvested broth cultures at 5,000gN for 20+-5 min.
 Remove the supernatant without disrupting the pellet. Re-suspend the pellet in 5-10 mL PBS.  Record resuspension volume.
 Prepare the final test suspension within 30 min of resuspending the culture.
 If necessary, disrupt the pellet using vortex-mixing or repetitive tapping/striking against a hard surface to disaggregate the pellet completely prior to re-suspending it in 10 mL. If necessary, add 1 mL of PBS to the pellet to aid in the disaggregation.
 If needed, dilute the 5-10 mL of resuspended culture in PBS to achieve a mean control carrier count level of 4.0-5.5 logs CFU/carrier for S. aureus and P. aeruginosa.
 Optical density/absorbance (e.g., 650 nm) may be used as a tool to monitor/adjust the diluted test suspension.
 Use the resuspended or diluted culture to prepare the final test suspension with the addition of the soil load.
 To obtain 500 uL of the final test suspension with the 3-part soil load, vortex-mix each component and combine in the following order using a calibrated micropipette:
 25 uL BSA stock
 35 uL yeast extract stock
 100 uL mucin stock
 Vortex soil suspension for 10s prior to adding microbial test suspension.
 340 uL microbial test suspension
 Briefly vortex the final test suspension with 3-part soil load (at room temperature, 21+-3°C) and use to inoculate carriers within 30 min of preparation.
 Streak inoculate an agar plate with a loopful of the final test suspension.  Incubate plate with the treated and control carrier plates and examine for purity after incubation at 36+-1°C for 72+-4 h.
 It is advisable to briefly rescreen each sterilized carrier for abnormalities prior to inoculation. Place carriers screened side up inside an empty, sterile plastic Petri dish (no more than 20 carriers/dish).
 Privacy curtain carriers have no backing material and may be inoculated on either side.
 Non-PVC and vinyl carriers are layered materials comprised of a smooth, colored top surface and a white fabric bottom; only the top surface will be inoculated.
 Vortex-mix the final test suspension for 10 s following the addition of the soil load and immediately prior to use.
 Inoculate the number of carriers required for the evaluation of the test substance (3 controls and 5 treated) along with a few extra carriers.
 Using a calibrated positive displacement pipette with a 10 uL tip, withdraw 10 uL of the final test suspension and deposit it at the center of each carrier (clean, screened and sterile); avoid contact of pipette tip with carrier and do not spread the final test suspension with the pipette tip.
 For consistency, vortex-mix the inoculum frequently during inoculation of the carrier set.
 The same pipette tip may be used to inoculate all carriers (unless the tip is compromised).
 Discard any inoculated carrier where the final test suspension has run over the edge.
 Transfer the Petri dish(es) with the inoculated carriers into a desiccation unit (with desiccant) and completely remove the lid of the Petri dish.  Close the desiccation unit door (or lid) and seal the unit. Apply vacuum to evacuate the desiccation unit.
 Note: do not exceed 40 inoculated carriers per desiccator to ensure carriers dry within the prescribed time.
 Maintain and monitor the vacuum level using a gauge. Achieve and maintain consistent level of vacuum (at 20-25 in of mercury, 508-635 torr, 677-847 mbar, or 68000-85000 Pascal) by leaving the vacuum on during the drying period with the desiccator stopcock opened or closed as necessary.
 Hold the inoculated carriers in the evacuated desiccation unit at 21+-3°C for 45 to 60 min. Visually inspect inoculated carriers to verify that they have completely dried and remove from desiccation unit. Do not use carriers that are visibly wet for testing.
 Record the time for all timed events.
 Depressurize the desiccator slowly to avoid the potential for carriers to move or flip.
 Use dried inoculated carriers for testing within 30 min following removal from desiccation unit; hold carriers in closed Petri dish at room temperature (21+-3°C) until use.
 Performance Assessment  -  Efficacy
 Evaluate 3 control carriers and 5 treated carriers for each test substance tested (one test organism and contact time /carrier type combination) unless specified otherwise.
 One set of control carriers per carrier type may be used for evaluating multiple test substances against one organism on one test day (assuming the carrier material, neutralizer, and soil load are the same).
 Using sterile forceps, transfer each dried carrier with the inoculated side up to a flat-bottom vial and cap the vial. Repeat until all carriers are transferred.
 Prepare the antimicrobial test substance. Use antimicrobial test substance within 3 hours of preparation or as specified by the manufacturer.
 In a timed fashion with appropriate intervals, sequentially deposit 50 uL of the test substance (equilibrated to 21+-3°C) with a calibrated micropipette over the dried inoculum on each test carrier, ensuring complete coverage.
 Note: Gently apply the antimicrobial test substance at a perpendicular angle to the inoculated carrier; do not forcefully deposit the disinfectant.
 Use a new tip for each carrier; do not touch the carrier surface with a pipette tip during the application of the test substance or the control substance; replace with new carrier(s) and vial(s) if this occurs.  Do not cap the vials.
 For non-foaming aerosols and pump/trigger spray products, obtain the test substance by dispensing the product into a sterile vessel for collection. Cap the vessel and use dispensed product within 30 min.
 For foaming spray formulations, allow the foam to break down for at least 5-10 minutes for the generation of a 1-2 mL liquid sample.  Cap the vessel and use dispensed product within 30 min.
 Do not process carriers where the test substance runs off the carrier or does not completely cover the inoculum spot; replace with new carrier(s) and vial(s) if this occurs.
 Conduct the test at room temperature (21+-3°C) for the selected contact time.  Use a certified timer to ensure that each carrier receives the required contact time.
 Process control carriers last. Each control carrier receives 50 uL PBS, equilibrated to 21+-3°C, instead of the test substance. Hold the control carriers for the same contact time as used for the test substance.
 Within +-5 s of the end of the contact period, add 10 mL of neutralizer equilibrated to 21+-3°C to each vial in the specified order according to the predetermined schedule.  Briefly vortex-mix (2-3 s) each vial following the addition of the neutralizer.
 For calculation purposes, the solution in the neutralized vial with carrier is considered to be 10[0] dilution.
 The neutralizer for the control carriers is the same as that for the treated carriers.
 Immediately following the addition of the neutralizer and briefly (2-3 s) vortex, allow carriers to sit in the vials for 5 minutes undisturbed then proceed as follows:
 Vortex-mix vials at high speed for 30 s (vortex-mix #1).
 Allow carriers to sit undisturbed in the vials for 5 minutes.
 Vortex-mix vials at high speed for 30 s (vortex-mix #2).
 Allow carriers to sit undisturbed in the vials for 5 minutes.
 Vortex-mix vials at high speed for 30 s (vortex-mix #3).
 Initiate dilutions within 30 min after neutralization and vortex-mixing. Initiate filtration within 30 min of preparing the dilutions.
 Dilute and filter samples from the treated and control carriers; process treated carriers first.
 Serially dilute the eluate from the 10[0] dilution prior to filtration by transferring 1 mL into 9 mL PBS in a dilution tube.
 Turn on vacuum and leave on for the duration of the filtration process.
 Prior to filtration, pre-wet each membrane filter with ~10 mL PBS.
 Use separate membrane filters for each eluate (neutralized solution); however, the same filtration unit may be used for processing eluates from a given carrier set starting with the most dilute sample first.
 Filter each sample through a separate 0.2 um PES membrane filter.
 For eluates from treated carriers remaining in the vial (10[0] dilution), vortex-mix the vial for ~5 s, carefully pour the eluate into the filter unit.
 If a carrier falls onto the filter membrane, aseptically remove it using sterile forceps.
 Rinse the treated vial with ~20 mL PBS, vortex-mix for ~5 s, pour the wash into the same filter unit.  For dilution tubes, rinse tube once with ~10 mL PBS, briefly vortex-mix, and pour into filter unit.
 Swirl the contents of the filter unit and quickly filter with limited pooling of liquid in the filter apparatus.
 Rinse the inside surface of the funnel unit with at least 20 mL PBS and filter the contents.
 Aseptically remove the membrane filter and place on the appropriate recovery medium.  Avoid trapping any air bubbles between the filter and the agar surface.
 Sterility controls.
 On the day of the test, filter ~20 mL of neutralizer and ~20 mL of the PBS used in the test using two separate membrane filters and place on TSA.
 Incubate these filters along with a plate of recovery medium (e.g., TSA) for 72+-4 h at 36+-1°C, record sterility results.
 Incubate plates at 36+-1°C for 48+-4 h for control carriers and for a minimum of 72+-4 h for treated carriers.
 Monitor filters daily to optimize counting of colonies. CFUs may be counted daily. Record controls after 48+-4 h and treated carriers after 72+-4 h.
 Count colonies and record results.
 Any level of contamination which interferes with the recording and interpretation of results will result in invalid data.
 For example, contamination occuring on multiple filters within one set of serial dilutions and/or across multiple carriers is considered systemic and the test is deemed invalid.
 For colony counts on filters in excess of 200 record as Too Numerous to Count (TNTC).
 If no colonies are present, record as zero.
 Report non-conforming data (e.g., systemic contamination and atypical serial dilution results) and repeat tests as necessary.
 Systemic contamination
 Atypical serial dilution results (e.g., higher CFUs at more dilute levels).
 Inspect the growth on the filters for purity and typical characteristics of the test microbe, see Attachment A, Table 1.
 If isolated colonies are present, assess one representative colony per 5-carrier set (treated) or 3-carrier set (controls) using a Gram stain.
 If confluent growth is present, perform a streak isolation on TSA or TSA with 5% sheep blood on growth taken from at least 1 carrier incubate at 36+-1°C for 24-48 h.
 If additional verification of the test organism is required, perform further confirmatory analyses (e.g., Vitek and biochemical analyses) and isolation streaks on selective media.
 Data Requirements
 Per test, use colony counts to determine log reduction.
 For an acceptable test, each of the three control carriers must exhibit counts between 4.0-5.5 logs CFU/carrier.
 Use values with at least three significant figures when performing calculations (e.g., log density, mean log density).  Report the final mean log reduction value with two significant figures (e.g., round up to the nearest tenth).
 Calculate the Colony Forming Units (CFU)/carrier using the following equation:  Log10i=1nYii=1nCixDixV,
where:
Y
                                       =
CFU per filter,
C
                                       =
volume filtered,
V
                                       =
total volume of neutralizer,
D
                                       =
10[-k],
k
                                       =
dilution,
n
                                       =
number of dilutions, and
i
                                      =
lower limit of summation (the fewest number of dilutions).
 When TNTC (Too Numerous To Count) values are observed for each dilution filtered, substitute 200 for the TNTC at the highest (most dilute) dilution and account for the dilution factor in the calculations.
 Calculate the log density of each carrier by taking the log10 of the density per carrier.
 Calculate the mean log10 density across treated carriers.
 Calculate the mean log10 density across control carriers.
 Calculate the log10 reduction (LR) for treated carriers: log10 reduction = the mean log10 density for control carriers minus the mean log10 density for treated carriers.
 For a set of treated carriers: when the 10[0] dilution (the contents of the vial with the carrier) is filtered either by itself or in addition to other dilutions and the data for each carrier result in zeros for each dilution filtered, report the LR as greater than or equal to the mean log10 density for the control carriers.
 Log reduction data based on estimates due to the occurrence of TNTC outcomes at each dilution in a dilution series for control and treated carriers is deemed unacceptable.

Attachment A
                      Preparation of Frozen Stock Culture

 Initiate new stock cultures from lyophilized cultures of Pseudomonas aeruginosa and Staphylococcus aureus from a reputable vender at least every 18 months.
          New frozen stock culture may be initiated one time using an existing, unexpired frozen stock culture as the source. Begin process at step 3 below, by streaking a loopful of the frozen stock culture onto 2 TSA plates.
 Open ampule of freeze-dried organism per manufacturer's instructions. Using a tube containing 5-6 mL of TSB, aseptically withdraw 0.5 to 1.0 mL and rehydrate the lyophilized culture. Aseptically transfer the entire rehydrated pellet back into the original tube of broth. Mix thoroughly. Incubate broth culture at 36+-1°C for 24+-2 h.
 At the end of the incubation timeframe, streak a loopful of the broth culture onto 2 TSA plates to obtain isolated colonies. Perform a streak isolation of the broth culture onto BAP as a purity check and streak the broth culture onto the appropriate selective media. Refer to appropriate selective media in Table 1.  Incubate all plates for 24+-2 h at 36+-1°C.
       Record results at the end of the incubation timeframe. Refer to Table 1 for results on selective media and diagnostic characteristics of the test microbes.
 From the TSA plates, select 3-5 isolated colonies of the test organism and re-suspend in 1 mL of TSB. For S. aureus, select only golden yellow colonies. For P. aeruginosa, select colonies from each of the two possible phenotypes present. Spread plate 0.1 mL of the suspension onto each of 6-10 TSA plates. Incubate the plates for 24+-2 h at 36+-1°C. If necessary, to obtain more frozen stock cultures, a larger suspension (e.g., 2 mL) may be prepared using the same ratio of TSB (1 mL) to number of colonies (3-5 colonies).
       Using the TSB suspension, perform a streak isolation of the suspension onto a BAP as a purity check, and streak on the appropriate selective media (refer to Table 1).
       Incubate all plates for 24+-2 h at 36+-1°C. Record results. Refer to Table 1 for results on selective media and diagnostic characteristics of the test microbes.
 After the incubation period, harvest growth from TSA plates by adding approximately 5 mL sterile cryoprotectant solution (TSB with 15% (v/v) glycerol) on the surface of each plate.  Re-suspend the growth in the cryoprotectant solution using a sterile spreader without damaging the agar surface.  Aspirate the suspension from the plate with a pipette and place it in a sterile vessel large enough to hold about 30 mL.
 Repeat the growth harvesting procedure with the remaining plates and continue adding the suspension to the vessel (more than 1 vessel may be used if necessary). Mix the contents of the vessel(s) thoroughly; if more than 1 vessel is used, pool the vessels prior to aliquoting culture.
 Immediately after mixing, dispense 0.5-1.0 mL aliquots of the harvested suspension into cryovials; these represent the frozen stock cultures.
          For QC purposes, perform a streak isolation of the pooled culture onto a BAP as a purity check and streak on appropriate selective media (refer to Table 1).
          Incubate all plates for 24+-2 h at 36+-1°C.
          Record results. Refer to Table 1 for results on selective media and diagnostic characteristics of the test microbes.
          After incubation, perform a Gram stain on growth from the BAP; observe the Gram reaction by using brightfield microscopy at 1000X magnification (oil immersion).
          Conduct confirmation using an automated identification system (i.e., Vitek) or biochemical and antigenic analyses from growth taken from the BAP according to the manufacturer's instructions.
 Store the cryovials at approximately -80°C for a maximum of 18 months. These cultures are single use only.
 If the characteristics of the organism are not consistent with the information in Table 1 at any step in the process, or the Vitek profile is inconsistent with the organism, discard the cultures and re-initiate the process.

Table 1. Selective media and diagnostic characteristics for P. aeruginosa and S. aureus
                                    Aspect
                                P. aeruginosa*
                                   S. aureus
                              Gram stain reaction
                                   Negative
                                   Positive
                     Mannitol Salt Agar
(Selective medium)
                                      N/A
       Circular, small, yellow colonies, agar turning fluorescent yellow
                       Cetrimide Agar
(Selective medium)
Circular, small, initially opaque, turning fluorescent green over time; agar fluorescent yellowish green
                                      N/A
                               Blood agar (BAP)
Flat, opaque to off-white, round spreading (1), metallic sheen, slightly beta hemolytic
         Small, circular, yellow or white, glistening, beta hemolytic

                      Typical Microscopic Characteristics
                                Cell appearance
Straight or slightly curved rods, single polar flagella, rods formed in chains; 0.5-1.0 um in diameter x 1.5-5.0 um in length
Spherical, occurring singly, in pairs and tetrads, sometimes forming irregular clusters; 0.5-1.0 um in diameter
      *After 24+-2 h (1) P. aeruginosa may display two phenotypes.

Attachment B
                             Neutralization Assay
The purpose of this section is to assess the effectiveness of the neutralization processes associated with this method.  Perform the neutralization assay with both microbes for each carrier type prior to testing to demonstrate the neutralizer's ability to inactivate the chemical.
Select a neutralizing medium that is not inhibitory to the test microbe. The acceptance criteria for acceptable neutralization is <= 50% difference in colony counts between the neutralization effectiveness, neutralization toxicity control, titer control, and carrier control.
 Refer to Section 3 in the preceding method for preparation of the test cultures. Conduct preliminary tests as necessary to determine appropriate dilution(s) of Test Suspension A (used to prepare Test Suspension B) to achieve the target challenge of 20-200 CFU per 10 uL or per carrier.
 Prepare Test Suspension A (without soil load). Serially dilute the microbial test suspension with PBS (e.g., through 10[-4] or 10[-5]). Select appropriate dilutions of Test Suspension A so that after the addition of the soil load, the Test Suspension B will achieve an average challenge of 20-200 CFU per 10 uL. Use Test Suspension A within 30 min of preparation.
 Prepare Test Suspension B (with soil load). Prepare the soil load: using a vortex, mix each component and combine 25 uL bovine serum albumin (BSA), 35 uL yeast extract, and 100 uL of mucin; then vortex-mix the solution. Combine 340 uL of diluted Test Suspension A and the 160 uL of the soil load (SL) and vortex-mix for 10 seconds.
 Ensure Test Suspension B provides an average challenge of 20-200 CFU per 10 uL.
 Two separate serial dilutions of Test Suspension A may be used to prepare two different concentrations of Test Suspension B to ensure at least one dilution with an average challenge of 20-200 CFU per 10 uL.
 A calibration curve (OD @ 650nm) may be used to estimate the number of viable organisms in Test Suspension A.
 Neutralization Treatments (see Attachment B, Figure 2)
 Treatment 1: Neutralizer Effectiveness. Add 50 uL of the test substance to each of three reaction vessels. At timed intervals, add 10 mL neutralizer to each vessel and briefly swirl (by hand). After 10 s, gently add 10 uL of neutralizer test suspension using a micropipette to each vessel and briefly vortex. Proceed with section 4).
 Treatment 2: Neutralizer Toxicity Control. Add 10 mL neutralizer to each of three reaction vessels.  At timed intervals, add 10 uL of Test Suspension B using a micropipette to each vessel and briefly vortex. Proceed with section 4).
 Treatment 3: Titer Control. Add 10 mL PBS to each of three reaction vessels.  At timed intervals, add 10 uL of Test Suspension B using a micropipette to each vessel and briefly vortex. Proceed with section 4).
 Treatment 4: Carrier Interference Control.  Add one carrier to each of three reaction vessels. At timed intervals, add 10 mL neutralizer to each vessel and briefly swirl (by hand).  After 10 s. gently add 10 uL of Test Suspension B using a micropipette to each vessel and briefly vortex.  Proceed with section 4).
 Hold the neutralization treatments for 10+-1 at room temperature (21+-3°C).
 At the conclusion of the holding period, vortex each reaction and filter each mixture through a separate, pre-wetted 0.2 um PES membrane filter.
 Wash each reaction vessel with ~20 mL PBS and vortex; filter the wash through the same filter membrane.  Finish the filtering process by rinsing the inside of the funnel unit with ~20 mL PBS and filter the rinsing liquid through the same filter membrane.
 Initiate filtration as soon as possible (e.g., within 30 min).
 Remove the membrane aseptically with sterile forceps and place it carefully over the surface of the recovery medium.  Avoid trapping air bubbles between the filter and the agar surface.
 Count and record CFUs daily, up to 72+-4 h.
                          Titer Interference Control
Add 50 uL of test substance to each vessel. At timed intervals add 10 mL neutralizer and swirl by hand.
                           Neutralizer Effectiveness
Add 10 uL of Test Suspension B to each vessel containing 50 uL test substance and 10 mL neutralizer.
Wait 10 s
Vortex and hold for 10 min at 21+-3°C. Proceed to vortexing/filtering.
Treatment 1
 50 uL test substance
Add 10 uL of Test Suspension B to each vessel containing 10 mL neutralizer.
                         Neutralizer Toxicity Control
Vortex and hold for 10 min at 21+-3°C. Proceed to vortexing/filtering.
Treatment 2
Add 10 uL of Test Suspension B to each vessel containing 10 mL PBS.
                                 Titer Control
Vortex and hold for 10 min at 21+-3°C. Proceed to vortexing/filtering.
Treatment 3
Add 1 sterile carrier to each vessel. At timed intervals, add 10 mL neutralizer and swirl by hand.
Treatment 4
Add 10 uL of Test Suspension B to each vessel containing the carrier and 10 mL neutralizer.
Wait 10 s
Vortex and hold for 10 min at 21+-3°C. Proceed to vortexing/filtering.
Figure 2: Neutralization Schematic


[Federal Register: October 29, 2008 (Volume 73, Number 210)]
[Notices]
[Page 64374-64376]
[DOCID:fr29oc08-101]

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SECURITIES AND EXCHANGE COMMISSION

[Release No. 34-58832; File No. SR-FINRA-2008-053]

Self-Regulatory Organizations; Financial Industry Regulatory
Authority, Inc.; Notice of Filing of Proposed Rule Change To Amend
Section 4(c) of Schedule A of the FINRA By-Laws To Increase Certain
Qualification Examination Fees

October 22, 2008.
    Pursuant to Section 19(b)(1) of the Securities Exchange Act of 1934
(``Act'') \1\ and Rule 19b-4 thereunder,\2\ notice is hereby given that
on October 15, 2008, Financial Industry Regulatory Authority, Inc.
(``FINRA'') (f/k/a National Association of Securities Dealers, Inc.
(``NASD'')) filed with the Securities and Exchange Commission (``SEC''
or ``Commission'') the proposed rule change as described in Items I,
II, and III below, which Items have been prepared by FINRA. The
Commission is publishing this notice to solicit comments on the
proposed rule change from interested persons.
---------------------------------------------------------------------------

    \1\ 15 U.S.C. 78s(b)(1).
    \2\ 17 CFR 240.19b-4.
---------------------------------------------------------------------------

I. Self-Regulatory Organization's Statement of the Terms of Substance
of the Proposed Rule Change

    FINRA is proposing to amend Section 4(c) of Schedule A to the FINRA
By-Laws (``Schedule A'') to increase certain qualification examination
fees. Below is the text of the proposed rule change. Proposed new
language is italicized; proposed deletions are in brackets.
* * * * *
    Schedule A to the By-Laws of the Corporation
* * * * *
Section 4--Fees
    (a) and (b) No change
    (c) The following fees shall be assessed to each individual who
registers to take an examination as described below. These fees are in
addition to the registration fee described in paragraph (b).

Series 4.............................  Registered Options Principal...........  [$80] $90
Series 6.............................  Investment Company Products/Variable     [$75] $85
                                        Contracts Representative.
Series 7.............................  General Securities Representative......  [$250] $265
Series 9.............................  General Securities Sales Supervisor--    [$60] $70
                                        Options Module.
Series 10............................  General Securities Sales Supervisor--    [$100] $110
                                        General Module.
Series 11............................  Assistant Representative--Order          [$60] $70
                                        Processing.
Series 14............................  Compliance Official....................  [$300] $320
Series 16............................  Supervisory Analyst....................  [$200] $210
Series 17............................  Limited Registered Representative......  [$65] $70
Series 22............................  Direct Participation Programs            [$75] $85
                                        Representative.
Series 23............................  General Securities Principal Sales       [$75] $85
                                        Supervisor Module.
Series 24............................  General Securities Principal...........  [$95] $105
Series 26............................  Investment Company Products/Variable     [$75] $85
                                        Contracts Principal.
Series 27............................  Financial and Operations Principal.....  [$95] $105
Series 28............................  Introducing Broker[/]-Dealer Financial   [$75] $85
                                        and Operations Principal.
Series 37............................  Canada Module of S7 (Options Required).  [$150] $160
Series 38............................  Canada Module of S7 (No Options          [$150] $160
                                        Required).
Series 39............................  Direct Participation Programs Principal  [$75] $80
Series 42............................  Registered Options Representative......  [$60] $65
Series 55............................  Limited Representative--Equity Trader..  [$85] $95
Series 62............................  Corporate Securities Limited             [$75] $80
                                        Representative.
Series 72............................  Government Securities Representative...  [$85] $95
Series 82............................  Limited Representative--Private          [$75] $80
                                        Securities Offering.
Series 86............................  Research Analyst--Analysis.............  [$150] $160
Series 87............................  Research Analyst--Regulatory...........  [$105] $115

    (1) through (3) No change
    (d) through (h) No change
* * * * *

II. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

    In its filing with the Commission, FINRA included statements
concerning the purpose of and basis for the proposed rule change and
discussed any comments it received on the proposed rule change. The
text of these statements may be examined at the places specified in
Item IV below. FINRA has prepared summaries, set forth in sections A,
B, and C below, of the most significant aspects of such statements.

A. Self-Regulatory Organization's Statement of the Purpose of, and
Statutory Basis for, the Proposed Rule Change

1. Purpose
    Any person associated with a member firm who is engaged in the
securities business of the firm must register with FINRA. As part of
the registration process, securities professionals must pass a
qualification examination to demonstrate competence in each area in
which they intend to work. These mandatory qualification examinations
cover a broad range of subjects on the markets, products, a person's
responsibilities in a given position, securities industry rules and the
regulatory structure. Some qualification examinations are sponsored
(i.e., developed) solely by FINRA while others are sponsored by the
Municipal Securities Rulemaking Board (``MSRB''), the North American
Securities Administrators Association (``NASAA''), the National Futures
Association (``NFA''), the Federal Deposit Insurance Corporation
(``FDIC''), other self-regulatory organizations (``SROs'') or jointly
among these entities.\3\
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    \3\ For example, FINRA administers and delivers the Series 6, 24
and 27 examinations, which are sponsored by FINRA. FINRA also
administers and delivers client-sponsored examinations, such as the
Series 9 and 10, which are sponsored jointly by several SROs (NYSE
Alternext U.S. LLC (``NYSE Alternext'') (formerly American Stock
Exchange), Chicago Board Options Exchange (``CBOE''), MSRB, FINRA,
NYSE Arca, Inc. (``NYSE Arca'') (formerly Pacific Stock Exchange,
Inc.) and NASDAQ OMX PHLX, Inc. (``Phlx'') (formerly Philadelphia
Stock Exchange)).
---------------------------------------------------------------------------

    FINRA administers qualification examinations via computer through
the PROCTOR[supreg] system \4\ at test centers operated by vendors
under contract with FINRA. FINRA charges an examination fee to
candidates for FINRA-sponsored and co-sponsored examinations. For
qualification examinations sponsored by a FINRA client and
administered/delivered by FINRA, FINRA charges a delivery fee that
represents either a portion of or the entire examination fee for a
particular examination.
---------------------------------------------------------------------------

    \4\ PROCTOR[supreg] is a technology system that supports
computer-based testing and training.
---------------------------------------------------------------------------

    The proposed rule change would amend Schedule A to increase certain
qualification examination fees.\5\ Each year, FINRA conducts a
comprehensive review of the examination fee structure, including an
analysis of the costs of developing, administering and delivering
qualification examinations. FINRA's 2008 review revealed that certain
operational costs have increased and will continue to increase over the
next few years. In particular, these costs consist of: (1) the cost of
providing the extensive network of test delivery centers; and (2)
technology costs required to maintain the PROCTOR[supreg] system. Based
on these findings, the proposed rule change would increase examination
fees with no single examination increasing more than $20. FINRA
believes that the proposed rule change would result in a better
alignment of program fees to associated operating costs.
---------------------------------------------------------------------------

    \5\ Schedule A sets forth examination fees for those
examinations that are sponsored or co-sponsored by FINRA and/or that
may be required by FINRA for its members.
---------------------------------------------------------------------------

    Specifically, the proposed rule change would amend Schedule A as
follows:

Series 4.............................  Registered Options Principal (Sponsored  From $80 to $90
                                        jointly by NYSE Alternext,CBOE, FINRA,
                                        NYSE Arca and Phlx).
Series 6.............................  Investment Company Products/Variable     From $75 to $85
                                        Contracts Representative.
Series 7.............................  General Securities Representative......  From $250 to $265
Series 9.............................  General Securities Sales Supervisor--    From $60 to $70
                                        Options Module (Sponsored jointly by
                                        NYSE Alternext, CBOE, FINRA, MSRB,
                                        NYSE Arca and Phlx).

Series 10............................  General Securities Sales Supervisor--    From $100 to $110
                                        General Module (Sponsored jointly by
                                        NYSE Alternext, CBOE, FINRA, MSRB,
                                        NYSE Arca and Phlx).
Series 11............................  Assistant Representative--Order          From $60 to $70
                                        Processing.
Series 14............................  Compliance Official....................  From $300 to $320
Series 16............................  Supervisory Analyst....................  From $200 to $210
Series 17............................  Limited Registered Representative......  From $65 to $70
Series 22............................  Direct Participation Programs            From $75 to $85
                                        Representative.
Series 23............................  General Securities Principal Sales       From $75 to $85
                                        Supervisor Module.
Series 24............................  General Securities Principal...........  From $95 to $105
Series 26............................  Investment Company Products/Variable      From $75 to $85
                                        Contracts Principal.
Series 27............................  Financial and Operations Principal.....  From $95 to $105
Series 28............................  Introducing Broker-Dealer Financial and  From $75 to $85
                                        Operations Principal.
Series 37............................  Canada Module of S7 (Options Required).  From $150 to $160
Series 38............................  Canada Module of S7 (No Options          From $150 to $160
                                        Required).
Series 39............................  Direct Participation Programs Principal  From $75 to $80
Series 42............................  Registered Options Representative......  From $60 to $65
Series 55............................  Limited Representative-Equity Trader...  From $85 to $95
Series 62............................  Corporate Securities Limited             From $75 to $80
                                        Representative.
Series 72............................  Government Securities Representative...  From $85 to $95
Series 82............................  Limited Representative--Private          From $75 to $80
                                        Securities Offering.
Series 86............................  Research Analyst--Analysis.............  From $150 to $160
Series 87............................  Research Analyst--Regulatory...........  From $105 to $115

    The effective date of the proposed rule change would be January 2,
2009. Specifically, the proposed examination fees would become
effective for ``120-day examination windows'' opened in the CRD[supreg]
on or after January 2, 2009.
2. Statutory Basis
    FINRA believes that the proposed rule change is consistent with the
provisions of Section 15A(b)(5) of the Act,\6\ which requires, among
other things, that FINRA rules provide for the equitable allocation of
reasonable dues, fees and other charges among members and issuers and
other persons using any facility or system that FINRA operates or
controls. FINRA believes that the proposed rule change to increase
certain qualification examination fees in the fee table in Schedule A
reflects FINRA's increased costs in developing, administering and
delivering qualification examinations.
---------------------------------------------------------------------------

    \6\ 15 U.S.C. 78o-3(b)(5).
---------------------------------------------------------------------------

B. Self-Regulatory Organization's Statement on Burden on Competition

    FINRA does not believe that the proposed rule change will result in
any burden on competition that is not necessary or appropriate in
furtherance of the purposes of the Act.

C. Self-Regulatory Organization's Statement on Comments on the Proposed
Rule Change Received From Members, Participants, or Others

    Written comments were neither solicited nor received.

III. Date of Effectiveness of the Proposed Rule Change and Timing for
Commission Action

    Within 35 days of the date of publication of this notice in the
Federal Register or within such longer period (i) as the Commission may
designate up to 90 days of such date if it finds such longer period to
be appropriate and publishes its reasons for so finding or (ii) as to
which the self-regulatory organization consents, the Commission will:
    (A) By order approve such proposed rule change, or
    (B) Institute proceedings to determine whether the proposed rule
change should be disapproved.

IV. Solicitation of Comments

    Interested persons are invited to submit written data, views, and
arguments concerning the foregoing, including whether the proposed rule
change is consistent with the Act. Comments may be submitted by any of
the following methods:

Electronic Comments

     Use the Commission's Internet comment form (http://
www.sec.gov/rules/sro.shtml); or
     Send an e-mail to<EMAIL_ADDRESS>Please include
File Number SR-FINRA-2008-053 on the subject line.

Paper Comments

     Send paper comments in triplicate to Secretary, Securities
and Exchange Commission, 100 F Street, NE., Washington, DC 20549-1090.

All submissions should refer to File Number SR-FINRA-2008-053. This
file number should be included on the subject line if e-mail is used.
To help the Commission process and review your comments more
efficiently, please use only one method. The Commission will post all
comments on the Commission's Internet Web site (http://www.sec.gov/
rules/sro.shtml). Copies of the submission, all subsequent amendments,
all written statements with respect to the proposed rule change that
are filed with the Commission, and all written communications relating
to the proposed rule change between the Commission and any person,
other than those that may be withheld from the public in accordance
with the provisions of 5 U.S.C. 552, will be available for inspection
and copying in the Commission's Public Reference Room, 100 F Street,
NE., Washington, DC 20549, on official business days between the hours
of 10 a.m. and 3 p.m. Copies of such filing also will be available for
inspection and copying at the principal office of FINRA. All comments
received will be posted without change; the Commission does not edit
personal identifying information from submissions. You should submit
only information that you wish to make available publicly. All
submissions should refer to File Number SR-FINRA-2008-053 and should be
submitted on or before November 19, 2008.

    For the Commission, by the Division of Trading and Markets,
pursuant to delegated authority.\7\
---------------------------------------------------------------------------

    \7\ 17 CFR 200.30-3(a)(12).
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Florence E. Harmon,
Acting Secretary.
[FR Doc. E8-25808 Filed 10-28-08; 8:45 am]

BILLING CODE 8011-01-P