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REPORT TO THE
MOBILE BAY ENFORCEMENT CONFERENCE
ON
BACTERIOLOGICAL CRITERIA FOR SHELLFISH
HARVESTING
by
MOBILE BAY TECHNICAL COMMITTEE
MOBILE BAY, ALABAMA
AUGUST 1972
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MOBILE BAY TECHNICAL COMMITTEE
Member
Robert F. Schneider, Chairman
W. F. Anderson
Arthur N. Beck
J. David Clem
James H. Coi1, Jr.
C. B. Kelly
F. James Silva
Representing
Environmental Protection Agency,
National Field Investigations
Center-Denver, Colorado.
Alabama Department of Conservation
and Natural Resources.
Alabama Water Improvement Commission
and Alabama State Health Department.
Department of Health, Education
and Welfare, Public Health Service,
Food and Drug Administration.
Scott Paper Company, Mobile, Alabama.
Environmental Protection Agency,
Water Supply Programs Division.
Environmental Protection Agency
Region IV, Atlanta, Georgia.
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TABLE OF CONTENTS
Page
MOBILE BAY TECHNICAL COMMITTEE 1
INTRODUCTION 3
THE MOBILE BAY SHELLFISH HARVEST PROBLEM 8
A SUBSTITUTE BACTERIOLOGICAL CRITERION 9
TECHNICAL COMMITTEE RECOMMENDATION 11
LITERATURE CITED 12
APPENDIX A
Request for the Establishment of the Mobile
Bay Technical Committee 13
APPENDIX B
Progress Report 28
APPENDIX C
Mobile Bay Technical Committee Meeting, Dauphin Island,
Alabama - March 2k, 1972 33
APPENDIX D
Correspondence Concerning a Proposed
Recommendation 56
APPENDIX E
Rapid Recovery of Escherichia coli from Estuarine
Waters 68
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INTRODUCTION
A Conference on the Matter of Pollution Affecting Shellfish in
Mobile Bay, Alabama was called by the Secretary of the Interior, and
was held in the City of Mobile on January 27-28, 1970. The Conference
Proceedingsl/contained a recommendation (No. 6) that emphasized
the need for reevaluation of the bacteriological criterion for
shellfish harvesting areas. The recommendation read: "In determin-
ing acceptability of areas in Mobile Bay for shellfish harvesting,
a more realistic criterion than the present method be employed to
more effectively evaluate the probability of pathogen presence. A
coordinated effort by the U. S. Department of Health, Education and
Welfare and the U. S. Department of the Interior, in cooperation with
the States, should be undertaken immediately to develop such a
criterion with respect to approval of areas for shellfish harvesting.
A technical committee shall be established by the Conferees to
assist in the development of such a criterion. This committee should
report to the Conferees no later than one year from the date of the first
session of conference."
After a lengthy delay, the Conference summary was approved;
on November 30, 1971 the EPA Enforcement Division, Region IV, Atlanta,
Georgia requested that the National Field Investigations Center-Denver
establish a technical committee in compliance with the Mobile Bay
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Enforcement Conference Recommendation No. 6. Subsequently, through
the Conferees the Technical Committee membership was established
in February, 1972. (Appendices A and B)
This report summarizes the considerations used by the Mobile
Bay Technical Committee for proposing a substitute bacteriological
criterion to evaluate the probability of pathogen presence in Mobile
Bay shellfish harvest areas.
HISTORICAL REVIEW OF THE COLIFORM BACTERIA CRITERION
The Mobile Bay Technical Committee reviewed the historical
and current rationale behind the present bacteriological criterion
for shellfish growing areas.
Near the turn of the century, investigators found that Escherichia
coli and other related bacteria normally occurred in the intestinal
tracks of mammals.— This assemblage of intestinal organisms became
known as the coliform bacteria group. Later, it was discovered
that the coliform bacteria were consistently present in sewage,
polluted water, and contaminated soil,— — — and the coliform
organisms became accepted as indicators of fecal pollution. Furthei—
more, it was found that water containing coliform bacteria often
contained enteric pathogens; thus the results of the "coliform test"
have become important considerations in judging the sanitary quality
of water in shellfish harvesting areas.
In 1936— , Dr. C. A. Perry proposed that Escherichia coli rather
than other coliform group bacteria be the index of pollution for both
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shellfish and shellfish waters. Five years later, a report titled,
"Public Health Aspects of Clamming in Raritan Bay" concluded that
"70 coliforms/100 ml, on the basis of average results from waters
overlying the shellfish beds, is recommended as the limiting allow-
able standards for the taking of hard clams to be eaten raw."
The 19^6 Manual of Recommended Practice for Sanitary Control
of the Shellfish IndustryZ/ presented the bacteriological criteria
for growing areas as follows: "The median bacteriological content
of samples of x^ater . . . shall not show the presence of organisms
i
of the coliform group [total coliform bacterial density] in excess
of 70/100 ml of water."
At the 5th and 6th National Shellfish Sanitation Workshops, par-
ticipants were in disagreement with the 70/100 ml criterion.
Studies—' 9/ had shown that the fecal coliform bacterial group was a
more sensitive index of fecal pollution than the total coliform
group. This conclusion was based on the fact that, in most instances,
the presence of fecal coliform organisms could be correlated with
the presence of fecal wastes from humans or other warm blooded
animals; conversely, investigators found that large numbers of non-
fecal coliforms could be recovered from waters and soils that were
remote from any known sources of warm blooded animal pollutants.
The Workshop participants reviewed these study findings and
discussed coliform standard for shellfish meats and a coliform
standard for the waters in which the shellfish are grown. An attempt
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was made by some participants to convince the Workshop that the
fecal coliform organisms were a better indicator of water pollution
than total coliform bacteria. Indicator levels of fecal coliform
bacteria were suggested for approved shellfish growing areas by
those who favored changing the bacteriological criterion. After
considerable consternation among the 1961* and 1968 Workshop repre-
sentatives, they recommended continued use of coliform organism
criteria and further study of the fecal coliform group in shell-
fish harvesting waters.
The substitution of the fecal coliform indicator group for the
total coliform group in judging shellfish harvest area sanitation
came under discussion again at the 7th National Shellfish Sanitation
Program (NSSP) in 1971.12/ During the Microbiology Task Force
deliberations at the Workshop, median fecal coliform values of 7-8
and 1*3 were recommended as growing area standards by representatives
of Maryland and Virginia, respectively. A spokesman from Lousiana
remarked that in his State a 23 fecal coliform MPN median standard
would close a high percent of "approved" shellfish-growing areas
which could not be shown to be affected by known sources of pollu-
tion. Delegates from Rhode Island proposed that the words "not
more than 10 percent of the samples ordinarily exceed an MPN of
230 per 100 ml" be omitted from the coliform standard. These actions
indicate the diversity of opinions expressed by scientists who were
actively trying to Devaluate bacteriological criteria for approved
shellfish-growing areas. After considerable discussion, all but two
State representatives in the 1971 NSSP Task Force voted to support
the following FDA recommendation:
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"The Food and Drug Administration (FDA) recommends to the 7th
National Workshop that the bacteriological criteria for shellfish
harvest areas be maintained as stated in the 19&5 Revision of the
NSSP Manual of Operations [not to exceed a median total coliform
value of 70/100 ml— ]. It is also recommended that the NSSP
continue to use the fecal coliform test as defined in Recommended
Procedures for the Examination of Seawater and Shellfish, APHA 1970,
as an adjunct to the coliform test to provide additional information
relative to water quality of shellfish harvest areas."
The FDA based this recommendation on five conclusions.
1. The present bacteriological criteria for approved growing areas
has been tested for many years and has demonstrated its effective-
ness in providing safe shellfish.
2. The 70/100 ml value of the coliform standard is well above the
lower limits of sensitivity of the multiple tube fermentation
procedure. Acceptable levels for a fecal coliform standard
would be in the range of the lower limits of sensitivity of the
multiple tube fermentation test resulting in more indeterminate
values of water quality data.
3. The coliform indicator group is a broad spectrum indicator of
pollution compared to the species of organisms recovered by the
fecal coliform test.
k. The relationship of numbers of fecal coliform organisms to direct
fecal sources has not been established.
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5. At the present time, there is no universal agreement among public
health experts as to the advisability of accepting a fecal
coliform growing area criterion for shellfish growing areas.
THE MOBILE BAY SHELLFISH HARVEST PROBLEM
The Proceedings of the Mobile Bay Enforcement Conference of 1970
contain a discussion of the problems affecting shellfish harvesting
in Mobile Bay.il/ It was reported that during the spring, flood
runoff (532,000 cfs) from the Tombigbee-Mobile River system trans-
i
ported large numbers of coliform bacteria into Mobile Bay. The
sanitary significance of these coliform densities was unknown;
yet, shell fishing areas in Mobile Bay were closed by the State of
Alabama during portions of 8 of the past 16 years because the bacterial
densities over the shellfish beds exceeded the 70/100 ml total coli-
form criterion for harvesting. The region has suffered substantial
economic losses because of these closures.
The problem was reiterated at a Mobile Bay Technical Committee
meeting at Dauphin Island, Alabama on March 2k, 1972 (Appendix C).
Committee spokesmen for the State of Alabama contended that the
application of the total coliform criterion unduly penalized the
Gulf Coast of the Southeastern Unived States. Coastal areas have a
potential for the occurrence of certain bacteria of the coliform group
in sources of no sanitary sgnificance without concomitant growth of
enteric pathogens. The Committee was reminded by representatives from
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the State of Alabama that during the past decade, portions of
Mobile Bay oyster reefs were closed on nine occasions for average
periods of 150 days. As of March 2^, 1972, a closure (effective
February k, 1972) resulted in a dockside economic loss of nearly
$100,000. This was equivalent to a total economic loss to the
Mobile area of almost $^00,000.
A SUBSTITUTE BACTERIOLOGICAL CRITERION
During the Technical Committee deliberations, opinions were
unanimous that a substitute bacteriological criterion for the total
coliform test was needed for evaluating shellfish harvest areas in
Mobile Bay. The Committee agreed that representatives of EPA and
FDA who are responsible for criteria development should consider the
following characteristics in the design of a substitute bacteriological
cri terion.
The substitute bacterial indicator should: (1) be more specific
for fecal pollution than the currently used indicator group; (2) stand
on its own merit as an indicator of actual sources of fecal pollution
without a dependency upon a specific relationship with the coli-
form group; (3) be more indicative of potential pathogen contamination;
(k) consistently indicate the sanitary quality of shellfish-growing
waters in diverse geographical locations; and (5) have a survival
time in the estuary consistent with that of bacterial pathogens
regardless of nutrient value of the water.
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After considering several avenues of approach, the Technical
Committee agreed that the two indicator tests most promising for
evaluating bacteriological contamination in Mobile Bay shellfish
harvest areas were (1) fecal coliform or (2) isolation of Escherichia
coli. Major benefits and weaknesses of these substitute criteria
were outlined by Committee members as follows:
(1) Presently, the fecal coliform test requires less time and
effort than the E_. col i measurement; therefore, results
can be obtained sooner, at less expense, and by fewer
microbiologists. However, studies by the EPA, FDA and
the State of Alabama have failed to demonstrate a consistent
total coliform/fecal coliform ratio, or determine a fecal
coliform value that can be directly related to known
sources of fecal pollution and consistently provide public
health protection to consumers of raw shellfish.
(2) Sanitary surveys and published documentation confirm that
the presence of E_. coli consistently indicates fecal
pollution and potential contamination by pathogenic
bacteria.
By letter to the Technical Committee on April 2k, 1972, the
Chairman proposed that the criterion for judging sanitary conditions
in Mobile Bay shellfish areas be based on the measurement of Escher ichia
coli (Appendix D). A majority of the Committee agreed with the
recommended E_. coli substitute criterion. Furthermore, the spokesman
for FDA cited a study on the rapid recovery of E_. col i from estuarine
waters (Appendix E). The study describes new methodology and concludes
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that £. col i can be isolated from raw seawater within 21* hours
without significant loss of accuracy. If the method accuracy can
be verified by collaborating studies, it will provide public health
protection to consumers of raw shellfish by rapidly and consistently
confirming fecal pollution and indicating potential contamination
by pathogenic bacteria.
TECHNICAL COMMITTEE RECOMMENDATION
In compliance with Recommendation No. 6 of the Mobile Bay
Enforcement Conference, the Technical Committee submits the follow-
ing statements to the Conferees: "To assist in the development of
a more realistic guideline than the present method for evaluating
the probability of pathogen presence in Mobile Bay, it is recommended
that the bacteriological criterion be based on the measurement of
Escherichia coli in waters overlying shellfish harvest areas. Further-
more, the Technical Committee recommends that the Environmental
Protection Agency, Food and Drug Administration and State shellfish
sanitation control agency combine their efforts to support the
research needed to develop a rapid, reliable method for measuring signifi'
cant numbers of E_. coli and to compare these values with other pollu-
tion indicator bacterial densities in estuarine waters used for
shellfish harvesting."
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LITERATURE CITED
1. Mobile Bay Enforcement Conference Proceedings.
2. Escherich, T. 1885. Die Darmbakterin des Neugeborenen und
Saughlings. Fortsch. Med. 3, 515 and 5^7.
3. Rogers, L. A., W. M. Clark, and A. C. Evans. 1915.
The characteristics of bacteria of the colon type found
in bovine feces. Jour. Infect. Dis. 15i 99.
4. Bards ley, D. A. 193**. The distribution and sanitary signi-
ficance of E. coli, B. lactus aerogenes and intermediate
types of coliform bacilli in water, soil, faeces and ice cream.
Jour. Hyg. 3**, 38.
5. Chen, C. C. and L. F. Rettger. 1920. A correlation study of the
colon-aerogenes group of bacteria with special reference
to the organisms occurring in the soil. Jour. Bact. 5, 253.
6. Perry, C. A. 1936. Examination of shellfish for fecal pollution.
Committee Report, APHA Yearbook 1935-1936, pp 111-117.
7. USPHS, 19^6. Manual of recommended practice for sanitary control
of the shellfish industry. U. S. Department of Health, Edu-
cation and Welfare, Washington, D.C.
8. Beck, W. J. 1964. Bacteriological criteria for shellfish
growing areas. Proc. 5th National Shellfish Sanitation Workshop.
9. Strebel, G. A. 1968. Coliform-fecal coliform bacteria in tidal
waters. Jour. San. Eng. Div., Proc. Amer. Soc. Civil Eng.
August 1968. pp. 6M-655.
10. Slanetz, L. W., C. H. Bartley, and K. W. Stanley. 1971.
Comparison of numbers of coliform and fecal coliform bacteria
in shellfish growing waters in relation to bacteriological
standards. Proc. 7th National Shellfish Sanitation Workshop.
11. USPHS, 1965. national shellfish, sanitation program, manual of
operations, Part III Public Health Service appraisal of state
shellfish sanitation programs. U. S. Department of Health,
Education and Welfare, Washington, D.C.
12. Gallagher, T. P., F. J. Silva, L. W. Olinger and R. A. Whatley.
1969. Pollution affecting shellfish harvesting in Mobile
Bay, Alabama. FWPCA, U. S. Department of the Interior, South-
east Water Laboratory, Athens, Georgia (mimeo).
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APPENDIX A
REQUEST FOR THE ESTABLISHMENT
OF THE
MOBILE BAY TECHNICAL COMMITTEE
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ENVIRONMENTAL PROTECTION AGENCY
Dalt: November 30, 1971
- Actine Director, Enforcement Division
Region IV
Subjut: Mobile Enforcement Conference Recommendation, In Re Shellfish
Technical Committee
To: Director, DFI-Denver Center
In accordance with our telephone conversation sometime back and
the follow-up conversation we had during our visit to Memphis
in September, I was directed by Mr. Stein to advise you that it
was your responsibility to see that recommendation 6 of the
enclosed conclusions and recommendations for the Mobile Bay
Enforcement Conference is followed.
As I indicated to you, when the project summary was approved on
March 2 of 1971 after a lengthy delay, the Regional Office had
moved to establish a technical committee in compliance with the
above referenced recommendation. However, in contacting personnel
in Mr. Stein's office, I was advised that this committee was to
be established at the national level a.nd that it would be
coordinated in Washington. Vic Lambou started working on this
but never got further than preliminary discussions with HEW.
After the standards section was removed from the Enforcement
Office, Lambou ceased working on the project.
Also as indicated to you, Jim Silva at my request prepared a
charter and developed a list of prospective committee members
(copy enclosed).
You will note in the recommendation that the committee is to
report to the conferees within a year. Because of the delay in
getting approval of the conference summary, the due date for the
committee's report is March 2, 1972.
o
Your early attention to this matter will be appreciated.
^
^1-i..-C-^—•
Enclosures: Conclusions & Recommendations
Charter & Member List
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YIRONMENTAL PROTECTION AGE
OFFICE OF ENFORCEMENT
DIVISION OF FIELD INVESTIGATIONS-DENVER CENTER
BUILDING 22 , ROOM 'MO , DENVER FEDERAL CENTER
DENVER. COLORADO 80225
10 : Mr. John White, Acting Director, DATE: December 6, 1971
Enforcement Division, Reg. IV, EPA
Atlanta
FROM : Director, DPI - Denver Center
SUBJECT: Establishment of Shellfish Technical Committee -
Mobile Bay
I have received your memorandum of November 30, 1971, requesting establish-
ment of a committee to recommend acceptable bacteriological criteria for
Mobile Bay. This request from your office is made to assure compliance
with Recommendation No. 6 of the Mobile Bay Enforcement Conference.
I am sure you recognize the delay in transmitting this request effectively
precludes completion of the required report by March 2, 1972. I am asking
Mr. Robert Schneider, Chief, Biology Branch, to serve as representative
from this office on the prospective coiiirni ttee. We have also requested
that Mr. David Clem, Director, Division of Shellfish Sanitation, serve as
the DHEW representative. At this time I would like to request the services
of Jim Silva of the Atlanta Regional Office, EPA, as the regional repre-
sentative. It is my intention that these people should meet at the
earliest mutual opportunity to designate an industrial and state repre-
sentative. Recommendations of this committee should be based upon con-
siderations relevant to Mobile Bay only. It is presently anticipated that
the committee will submit a progress report to the Conferees by March 1972.
Your early attention to this proposal will be appreciated and we look forward
to working with you and your excellent staff in completing this assignment in
a timely manner. You will be kept fully informed of committee activities.
If you have any further questions, please do not hesitate to contact me.
Thomas P. Gallagher
cc: Adm. , Reg. IV, EPA
Dir., Enf. Proceedings Div., EPA, w/cy
memo, Act. Dir., Enf. Div., Reg. IV
11/30/71 , same subject
Chief, Bio. Br., DPI, w/memo 6 encl
Dir., Div. of Shellfish San., DHEW
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22 'llO
December 6, 1971
Mr. David Clem, Director
Division of Shellfish Sanitation
DHEW, 200 "C" St., S.V,'.
Washington, D. C. 2020*i
.Dear Mr. Clem:
As you know, Recommendation Ho. 6 of the Mobile Boy Enforcement
Conference directed that a technical committee be established to
develop a more realistic criterion than the present method to nore
effectively evaluate the probability of pathogen presence in Mobile
Bay. This responsibility has been assigned to Division of Field
Investigations Center - Denver, EPA.
I have designated Mr. Robert Schneider as this Division's repre-
sentative on this corrr.'ii ttee and I would like to solicit you as a
representative also. I have asked Mr. Schneider to be in touch with
you to arrange a rceeting at the earliest convenience. A copy of my
letter to the Regional Office is also enclosed. I believe that your
participation on this committee is absolutely essential to the de-
velopment- of relevant recommendations to the Conferees.
Your early attention to this matter v/Ill be greatly appreciated.
Sincerely,
Thomas P. Gallagher
Di rector
encl. (1)
Cy memo 12/6/71 to Act. Dir.
Enf. Div., Reg. IV
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DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
PUBLIC HEALTH SERVICE
FOOD AND DRUG ADMINISTRATION
WASHINGTON. D.C. 20204
December 14, 1971
Mr. Thomas P. Gallagher
Director
Environmental Protection Agency
Office of Enforcement
Division of Field Investigations
Building 22, Room 410
Denver, Colorado 80225
Dear Tom:
This replies to your letter of December 6 regarding the
establishment of a technical committee to consider recommendation
No. 6 of the Mobile Bay Enforcement Conference. I would be
pleased to participate on this committee and to attend a
meeting of the committee in early 1972. 1 would also like
to request that Mr. Daniel A. Hunt, Assistant Director, be
appointed as my alternate to the committee.
I will look forward to hearing from you or Mr. Robert Schneider
regarding the time and place of the meeting.
Sincerely yours,
J. David Clem
Director
Division of Shellfish Sanitation (BF-230)
Office of Food Sanitation
Bureau of Foods
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ENVIRONMENTAL PROTECTION AGEN'"'
i
Date: December 23, 1971
Reply to Director, Enforcement Division
Ann of: Region !v
Subject: Establishment of Shellfish Technical Committee--Mobile Bay
To: Director, DFI - Denver Center
The proposal set forth in your memorandum of December 6 concerning the
above subject is satisfactory.
Mr. Traina has authorized Mr. Jim Silva to serve as regional representa-
tive on the committee.
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NVIRONMENTAL PROTECTION AGE^ ^Y
OFFICE OF ENFORCEMENT
DIVISION OF FIELD INVESTIGATIONS-DENVER CENTER
BUILDING 22 • ROOM ^10 • DENVER FEDERAL CENTER
DENVER, COLORADO 80225
k January 1972
J. David Clem, Director
Division of Shellfish Sanitation
Office of Food Sanitation
Bureau of Foods
Washington, D.C.
Dear Dave:
The first meeting of the Technical Committee to consider
recommendation No; 6 of the Mobile Bay Enforcement Conference
wi11 be held in:
Environmental Protection Agency
Region IV Office
3rd Floor
1A21 Peachtree Street, NE
Atlanta, Georgia
We will meet in Mr. Jim Silva's office at 1:00 p.m. on
Tuesday, January 18, 1972. I have prepared a charge that
we will discuss at this first meeting.
1 will look forward to seeing you in Atlanta.
Sincerely,
Robert F. Schneider, Acting Chief
Biology Branch
RFS:ls
cc: T. P. Gallagher, Director, DFI-DC
F. J. Silva, Atlanta Regional Office
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ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF ENFORCEMENT
DIVISION OF FIELD INVESTIGATIONS-DENVER CENTER
BUILDING 22 , ROOM ^10 , DENVER FEDERAL CENTER
DENVER, COLORADO 80225
COPY
10 : Mr. F. J. Silva DATE: 12 January 1972
Atlanta Regional Office
FROM : Chief, Biology Section
SUBJECT: Postponement of Technical Committee to Study Mobile Bay Enforcement
Conference Recommendation No. 6.
Following our telephone conversation of January 10, 1972 I contacted
Mr. Clem to advise him of the need to postpone our Technical Committee
Meeting until February. He has arranged his calendar accordingly.
As it is set up now, we will meet with you in your Atlanta Office
at 1:00 p.m. on Tuesday, February 1, 1972. There will be five of
us at this first meeting as follows:
Comm i 11ee Membe r s
Robert Schneider - DFI-DC
Jim Silva - Atlanta Region - EPA
Dave Clem - FDA Headquarters
Others
Bobby Carroll - SEWL - EPA
Roger Olmsted - Atlanta Region - FDA
Thanks for letting me know of your January calendar conflict. I am
looking forward to our February meeting.
R. F. Schneider
RFS/lms
cc: T. P. Gallagher, DFI-DC
J. D. Clem, FDA, Washington, D.C.
B. J. Carroll, SEWL, Athens, GA
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MEMORANDUM DEPARTMENT OF HEALTH, ED' VTION, AND WELFARE
PUBLIC HEALTH SERVICE
FOOD AND DRUG ADMINISTRATION
Regional Food ft Drug Director
10 : (ATL-F1) DATE: January 17, 1972
ATTN: Chief, Special Programs Branch (ATL-F15)
Director
ROM : Division of Shellfish Sanitation (BF-230)
Office of Food Sanitation/Bureau of Foods
Jlill^CT1
First. Meeting of the Technical Committee on Recommendation 6
of the Mobile Bay Enforcement Conference
1. This is to confirm my plans to visit the Regional
Office, February 1, in connection with a meeting of the
technical committee organized to consider recommendation
No. 6 of the Mobile Bay Enforcement Conference. I suggest
that Mr. Roger Olmsted accompany me to the meeting that
has been scheduled for 1:00 p.m. in Mr. Jim Silva's office.
I plan to visit your office, opening of business, Feb. 1.
JX David Clem
cc= R. Schneider, EPA, Colorado
T. Gallagher, DFI-DC
F. Silva, ATL Regional Office
R. Dimsted, ATL Regional Office
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I
ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF ENFORCEMENT
DIVISION OF FIELD INVESTIGATIONS-DENVER CENTER
BUILDING 22 . ROOM /no , DENVER FEDERAL CENTER
DENVER. COLORADO 80225
10 February 1972
Mr. James McDermott
1921 Jefferson Davis Highway
Arlington, Virginia 202^2
Dear Mr. McDermott:
Through the Environmental Protection Agency - Office of Enforcement,
I have been requested to establish a committee to recommend acceptable
bacteriological criteria for Mobile Bay shellfish harvesting areas.
I would Hke to solicit the services of Mr. C. B. Kelly of your staff
to join this committee as a representative for Water Hygiene.
As a way of giving you some background, I will briefly describe the
history behind this request. On January 27 and 28, 1970 a Conference
called by the Secretary of the Interior was held in Mobile, Alabama
concerning the economic effects of pollution in Mobile Bay upon the
Alabama shellfish industry. Recommendation No. 6 of the "Proceedings--
Conference in the Matter of Pollution Affecting Shellfish Harvesting
in Mobile Bay, Alabama" deals with shellfish growing water criteria.
The recommendation reads as follows: "In determining acceptability
of areas in Mobile Bay for shellfish harvesting, a more realistic
criterion than the present methods [total coliform density of 70 per
100 ml] be employed to more effectively evaluate the probability of
pathogen presence. A coordinated effort by the U. S. Department of
Health, Education and Welfare and the U. S. Department of the Interior
(now E.P.A. responsibility) in cooperation with the states should be
undertaken immediately to develop such a criterion with respect to
approval of areas for shellfish harvesting. A technical committee
shall be established by the Conferees to assist in the development
of such a criterion."
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Page 2
Mr. James McDermott
Arlington, Virginia
10 February 1972
Currently, the technical committee membership consists of: Robert F.
Schneider - Committee Chairman; F. James Silva - E.P.A. Regional
Representative; J. David Clem - DHEW Representative.
At a recent committee organization meeting in Atlanta, Georgia
Mr. Kelly's name was suggested as a committee member to represent
Water Hygiene. Because of his vast experience in microbiology and
knowledge of shellfish problems I believe his participation on the
technical committee is essential to successful development of relevant
recommendations to the Conferees.
Arrangements are being made at the Water Hygiene Dauphin Island
facility in Mobile, Alabama for a technical committee meeting place.
The first meeting of the technical committee is scheduled for March
23i 1972. All committee members will be provided with a specific
schedule as soon as arrangements are final.
Your early attention to this matter will be greatly appreciated.
Sincerely,
Robert F. Schneider
Chief, Biology Branch
RFS/ls
be: Mr. Kelly
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ENVIRONMENTAL PROTECTION AGENCY
WASHINGTON, D.C. 20460
MR 2 - 1972
Mr. Robert F. Schneider
Chief, Biology Branch
Division of Field Investigations — Denver Center
Office of Enforcement
Environmental Protection Agency
Denver, Colorado
Dear Mr. Schneider:
I shall be pleased to assign Mr. C. B. Kelly to serve on the
Technical Committee to develop criteria of quality for shellfish
growing areas. He will be prepared to attend a meeting of the
committee to be held at Dauphin Island, Alabama on March 24, 1972
Sincerely yours,
James H. McDermott, P.E.
Director
Water Supply Programs Division
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22 410
15 February 1972
Mr. C. B. Kelly
Special Assistant for Research
Division of Criterion Standards
Uater Supply Program Division
fcoom 503. Crystal 1^11 Buildin.R 2
Washington, D.C. 20460
Dear Mr. Kelly:
On behalf of the Technical Committee I would like to velcone you
as a member. I an looking forward to eeeting with you and others
of the Technical Coraeftttee to disenss our task of developing a
Eoro realistic criterion for shellfish growing areas in Mobile,
Alabana.
The Technical Coranittee membership is as follows:
1) J. David Clem, PDA Representative;
2) F. Janes Silva, EPA Regional ReprcBcntative;
3) Arthur Bock, Alabama Water Improvement and Health
Departncint Repreaentat ivc;
4) James Coil, Pulp and Paper Industry Representative; and
5) VT. B. Anderson, Alabana Conservation and Natural
ResourccB Representative.
A Technical Corsnittea neetln^ in bcln?, planned and will be held
at the E.P.A. Kater Hygiene Laboratory, Dauphin Island, Mobile,
Alabama on Itorch 24, 1972. You will be provided with a specific
standard when arrangements are final.
Sincerely,
Robert F. Schneider
Chief, Biology Branch
RFS/ls
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22 410
February 1972
Mr. Arthur 11. Brick, Technical Secretary
Water Improvement Centals s ion
State Office Building
Ifontgoiaery, Alabaua 3G104
Dear JJr. Beck:
On behalf of the Technical Copsnittee I would lite to wclcarui
you SB a cessbor. I as loaltiag forward to neccln^ with you aad
others of the Technical Contralttac to discucs o-.ir task of
developing a eoro realistic criteria for shellfish growing
areas la libblle Bay.
A Technical CcKnrf.ttee tisctinq Is boinj; planned and vill be
held in the E. ?. A. Water Hygiene laboratory. Dauphin Island,
Mobile, Alahana on March 24, 1972. You will be provided with a
specific schedule &G soon as arrsugerasnts are final.
Sincerely,
Robert F. Schneider
Chief, Biology Branch
BFS/la
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22 410
14 February 1972
Hr. James Coil
lianager of Adainistrative Services
Scott Paper Conpany
P. 0. Box 2447
Mobile, Alabama 36601
Dear Hr. Coll:
Pursuant to our telephone conversation of February 11, 1972 rsay
I take this opportunity to welcoTjO you to the Mobile Bay Tech-
nical Coiarcittce. I an looking forward to meeting with you and
others of tho Ccraaittae to discusc onr challenging task cf
rcconasendlng acceptable bacteriological criteria for shellfifih
areas in Mobile Bay.
A Technical Cornaitteo Kocting is beinR plamied and will be held
at the S. P. A. Wattr Hygiene Laboratory, Dauphin Island, Mobile,
Alabana on March 2h, 1972. You will be provided Kith specifics
as soon as &rrangeaent8 arc final.
Sincerely,
Robert F. Schneider
Chief, Biology Branch
RFS/lo
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APPENDIX B
PROGRESS REPORT
-28-
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PROGRESS REPORT
MOBILE BAY TECHNICAL COMMITTEE
R. F. Schneider
On January 27-28, 1970 in the City of Mobile, Alabama, a
Conference in the Matter of Pollution Affecting Shellfish in Mobile
Bay, Alabama was called by the Secretary of the Interior. Recommen-
dations contained in the Conference Proceedings included one that
emphasized reevaluation of the bacteriological criterion for shell-
fish harvesting areas.
Recommendation No. 6 reads: "In determining acceptability
of areas in Mobile Bay for shellfish harvesting, a more realistic
criterion than the present method be employed to more effectively
evaluate the probability of pathogen presence. A coordinated effort
by the U. S. Department of Health, Education and Welfare and the U. S.
Department of the Interior, in cooperation with the States, should be
undertaken immediately to develop such a criterion with respect to
approval of areas for shellfish harvesting. A technical committee
shall be established by the Conferees to assist in the development of
such a criterion. This committee should report to the Conferees no
later than one year from the date of the first session of conference."
In compliance with Recommendation No. 6 of the Mobile Bay Enforce-
ment Conference the following persons agreed to serve on the Technical
Committee: Mr. Robert Schneider, NFIC-Denver
Mr. J. David Clem, DHEW, Washington, D.C.
Mr. F. James Silva, EPA, Atlanta, Georgia
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On February 1, 1972, the members listed above met in Atlanta,
Georgia with Mr. Bobby Carroll, Chief Bacteriologist, EPA, Southeastern
U. S. Region and Mr. Roger Olmstead, FDA, Southeastern U. S. Region.
This meeting had a multi-purpose: (1) recommend prospective representa-
tives from the pulp and paper industry, fishery industry and the State
of Alabama; (2) review the highlights of the 1970 Mobile Bay Enforcement
Conference; (3) discuss the rationale behind the present bacteriological
criterion for shellfish growing areas and (1») jointly review the charge
to the technical committee.
Suggested additions to the Technical Committee membership included:
Mr. C. B. Kelly of EPA to represent the Water Hygiene Program; Mr. James
Coil of Scott Paper Company to represent the pulp and paper industry;
Mr. Arthur Beck, the Chief Sanitary Engineer of Alabama to represent
the Alabama Water Improvement Commission and the Alabama Health Depart-
ment; and Mr. W. F. Anderson, Chief of the Alabama Marine Resources to
represent the Alabama Department of Conservation and Natural Resources.
These individuals have agreed to serve on the Technical Committee in
the capacity noted above.
Highlights of the 1970 Enforcement Conference Proceeding and the
rationale behind the total coliform standard for shellfish growing
areas were reviewed by attendees at the meeting in Atlanta, Georgia on
February I, 1972. Discussions centered around tne fact that during tne
late winter or spring, flood runoff from the Tombigbee-Mobile River
system transports large coliform concentrations into Mobile Bay. The
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sanitary significance of these total coliform densities is unknown; yet,
shellfishing areas in Mobile Bay have been closed by the State of Alabama
because the bacterial densities over the shellfish beds exceeded the
70/100 ml total coliform standard for harvesting. The region has suffered
substantial economic.losses because of these closures.
The most recent closure is in effect, curreitly. On January 28,
1972 the State of Alabama closed all areas of Mobile Bay east of Dauphin
Island Bridge because the total coliform density exceeded the 70/100 ml
criterion. This closure means that some of the most productive shell-
fish areas of the Bay (Bon Secour Bay) are closed to harvesting for at
least a portion of the 1972 season. Economic loss to the region because
of past closures (excluding the 1972 closure) is estimated to exceed
200 thousand dollars.
After reviewing and discussing Recommendation No. 6 and other
shellfish related problems in Mobile Bay the following charge to the
Technical Committee was developed:
"The Technical Committee is requested to assist in the development
•
of a more realistic bacteriological criterion than the present method
[total coliform density of 70/100 ml] for determining the probability
of pathogen presence in Mobile Bay shellfish harvesting waters. To
accomplish this goal the Committee may form such sub-committees as
needed to review various avenues of approach, provide technical infor-
mation or conduct studies. The Committee will select an approach and
will set into motion a scheduled program to fulfill its charge. Further-
more, the Committee may exercise such latitude as required in fulfil ling
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its charge and shall meet at such times and places as necessary."
In response to this charge the Technical Committee has agreed to
meet in Mobile, Alabama on March 2*t, 1972. Various bacteriological
criteria for determining the acceptability of Mobile Bay waters for
shellfish harvesting will be discussed. Consideration will be given to
the sanitary significance of fecal coliform, fecal streptococci,
FC/FS ratio, or other parameters that could be used for evaluating
shellfish harvesting waters. The Committee will appoint a subcommittee
to review technical reports concerning the "State of the Art" and
sanitary significance of the various bacteriological criteria. The
subcommittee should be prepared to submit a written report of its
findings to the Technical Committee by August 1, 1972. After review-
ing the subcommittee findings, the Technical Committee will meet at
their earliest opportunity (no later than September 15, 1972) to formu-
late a recommendation. The Committee chairman will prepare and submit
the final recommendation to the Conferees by October 1, 1972.
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APPENDIX C
MOBILE BAY TECHNICAL COMMITTEE MEETING
DAUPHIN ISLAND, ALABAMA
MARCH 2k, 1972
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22
I March 1972
Mr. James Coil
Manager of Administrative Services
Scott Paper Company
P.O. Box 2^7
Mobile, Alabama 36601
Dear Mr. Coll:
The technical conijnltteo to consider recommendation no. 6 of the
Mobile Bay Enforcement Conference will r.eet in Mobile, Alabama
on March 2*1, 1972. The meeting Is scheduled for H:00 a.m. In
the library (conference room) of the V.'ater Hycilene laboratory
located on Dauphin Island. If you need specific directions,
please contact: Richard J. Hammerstro:n, Director
Gulf Coast Water Supply Laboratory
EPA
P.O. Box 15-3
Dauphin island, Alabama 3&52S
Phone: 205/861-2962 (not an FTS number)
At the meetInn we plan to discuss several bacteriological criteria
for determining the acceptability of Mobile Bay waters for shell-
fish harvesting. It v«uld be helpful to the committee If you
would present your views on the use of fecal coliform tests (tempera-
ture elevated bacteria) versus E. col I (IMVIC) tests or other alter-
nate methods to determine sanitary conditions In shellfish harvest-
Ing areas.
I am looking forward to x^orklng with you on this technical
committee.
Sincerely,
Robert F. Schneider
Chief, Biology Branch
RFS/ls
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22
I March 1972
Mr. J. David Clean
Division of Shellfish Sanitation
Bureau of Foods
Washington, D.C.
Dear Dave:
Plans have been finalized for our technical committee meeting.
We will meet at the Water Hygiene Laboratory on Dauphin Island
at 9:00 a.m. on March 2-'», 1972.
Because four of the committee members were absent from the
Atlanta meeting, there wl11 be a need for repeating some of
our previous discussions. Would you discuss the rationale
behind the present bacteriological criterion for shellfish
growing areas as you did for us in Atlanta?
I will be looking forward to working with you again on March 2*
Sincerely,
Robert F. Schneider
Chief, Biology Branch
RFS/ls
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22
1 March 1972
Mr. F. J. Sllva
EPA, Region IV
Suite 300
U21 Pcachtroe Street, NE
Atlanta, Scorpio 30309
Dear Jim:
Our technical meeting is set for 9:00 a.m. on March 2h at the
Water Hygiene Laboratory, Dauphin Island, Alolxv-ia. Thinks for
all your special efforts as I mentioned in our recent telephone
conversation. I do have one more favor to ask of you. Because
four of the coffinIttee nnnbers were absent fron the Atlanta
meeting, there will be a need for repeating some of our previous
discussions, l.'ould you review the highlights of the 1970
Mobile ?ay Enforcement Conference? This would 31 ve the ricmbers
of our full connittee a common background.
I'll be looking forvwrd to our Dauphin Island meeting \vith the
opportunity to work with you on tills long overdue shellfish
standard revision.
Sincerely,
Robert F. Schneider
Chief, Blolociy Branch
RFS/ls
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22
1 March 1972
Mr. Arthur H. Beck, Technical Secretary
Water Improvement Co-Tirclsslon
State Office Building
Montgomery, Alabaraa 3610*»
Dear Mr. Deck:
The technical committee to consider recommendation no. 6 of the
Mobile !3ay Enforcement Conference will meet In Mobile, Alabama
on March 24, 1372. The meeting is scheduled for 9:00 a.m. In
tho library (conference room) of the l/ater liyglcnc laboratory
located on Dauphin Island. If you need specific directions please
contact: Richard J. l-te-r.ierstro.T?, Director
Gulf Coast Water Supply Laboratory
EPA
P.O. Box 153
Dauphin island, Alabama 36528
Phone: 205/361-2962 (not an FTS number)
Some of the corral ttee members are not fatal liar with the recent
closure of Mobile Bay. At our March 24 meeting would you brief
the controlttee on the Bay closure and discuss the State of Alabama
views on the sanitary significance of the present bacteriological
criterion?
I asi looking forward to working with you on this special technical
committee.
Sincerely,
Robert F. Schneider
Chief, Biology Branch
RFS/ils
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22 410
1 March 1572
Mr. C. B. Kelly
Special Assistant for jJese.irch
Division of Criterion Standards
Water Supply Program Division
ROOM 509, Crystal ttall Suilulnn 2
Washington O.C. 20'»GO
Dear Mr. Kclly:
The technical convnlttee to consider rcconricntlntlon no. 6 of the
Mobile Bay Enforcement Conference will .-.icct in .Mobile, Alabama
on March 2'j, 137-2. The Kieetinq Is scheduled for 3:^0 a.m. In
tho library (conference roo-~>) of the. Water Hygiene laboratory
located on Dauphin Island, if yo:i nccsd specific directions
please contact: Richard J. Narwujrstrom, Director
Gulf Coost V/ator Supply Laboratory
EPA
P.O. Cox 1!#
Douphln Island, Alabama 36523
Phone: 205/561-2562 (not an FT5 number)
At the. laeetln? \:o. pl«5n to discuss several bacteriol or, lea I criteria
for determining the acceptability of Mobile Say waters for shell-
fish harvesting, it would be helpful to th& coi.TU:!ttGc If you
would present your views on the sanitary stqnlfIcanee of various
bacteriological indicators such as fecal collforni.
I am looking forward to meeting you and having the opportunity
to worlclaogcther on developing a rccorraenciatlon for the Conferees.
Sincerely,
Robert F. Schneider
Chief, Biology Oranch
RFS/ls
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22 410
2 March 1572
Mr. W. G. Anderson
Department of Conservation and Natural
Resources
Marine Resources Division
P.O. Box 186
Dauphin Island, Alabama 36523
Dear Mr. Anderson:
The technical corns taca to consider recoracndatlcn no. 6 of the
Mobile Bay Enforcement Conference will fncet In Mobile, Alabama on
March ?.•'», 19/2. Th2 meeting Is schctJuled for 5:00 a.ci. In the
library (conference! room) of the Water Hygiene laboratory located
on Dauphin Island, tf you need specific directions please contact:
Richard J. Mannerstram, Director
Gulf Coast Water Supply Laboratory
EPA
P.O. Box 158
O.iuphfn Island, Alabama 36523
Phone: 205/2&1-22&2 (not an FTS number)
Recently, 1 lesrneJ that Mobile Day Is closed to shellfish harvest-
ing again this season. It would be helpful to the co~rclttce If
you would present soi:»e Inforrnatlon concerning the economic loss or
other problems to the Mobile area because of this closure.
I'n lookInq forward to having the opportunity of working with you
on this technical committee.
Sincerely,
Robert F. Schneider
Chief, Biology Sranch
RFS/ls
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State of Alabama
Department of Public Health
State Office Building
Montgomery, Alabama
IRA L. MYERS, M. D.
STATE HEALTH OFFICER
March 6, 1972
Mr. Robert F. Schneider
Chief, Biology Branch
Environmental Protection Agency
Water Quality Office
Bldg. 22, Room 410
Denver Federal Center
Denver, Colorado 80225
Dear Mr. Schneider:
Thank you for your letter of March 1, 1972 advising me of
the meeting date and place of the newly formed Technical Committee to
implement the recommendations of No. 6 of the Mobile Bay Enforcement
Conference.
I will be happy to make any contribution that I can to the
Committee members at the March 24th. meeting.
Yours ver
ANB/iw
fief Engineer & Director
Bureau of Environmental Health
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Status of Bacteriological Criteria
for Shellfish Areas
The fecal coliform indicator group has been under
consideration by the National Shellfish Sanitation Program
(NSSP) .as a substitute for the coliform group in the
bactcr.iologicn] criteria for shellfish-growing areas for a
number of years. A fecal coliform standard of 7.8 was
proposed at the Fifth National Workshop in 1964 which
*
represented a coliform/fccal coliform ratio of approximately
10/1 when compared to the present coliform standard. There
appeared to be an assumption by the Fifth Workshop that any
new criteria or standard must have a consistent correlation
with the existing coliform standard. The proposed standard
was rejected on the basis of insufficient data and additional
studies were recommended.
The Microbiology Task Force of the Sixth National Workshop
again concluded that there was insufficient evidence for
recommending a fecal coliform standard and requested that
further work be done. .In response to this request, shellfish
sanitation personnel hnvc analyzed growing area data from
Louisiana, Mississippi, Alabama, Florida, Georgia, Virginia,
New York, Rhode Island, Massachusetts, and Maine. These data
Prep;M-cIl~Fy~ tiTe" Division of Shellfish Sanitation, Rurc.iu of Foods,
Food and Dru«> Administration, Washington, D. C. 20204 for prcsenta
tion ;it Mobile; Hay Technical Committee Meeting, Dauphin Island,
Al.-ibamu, March 21, 1972.
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2
indicate that there is no consistent coliform/fecal coliform
.relationship in cstuarine waters. The coliform/fecal coliform
ratio was found to vary from 1/1 to 10/1 depending upon such
factors as geographic location, meteorological conditions,
season, pollution source, distance of sampling station from
pollution source (dilution effect) and salinity, nutrients,
or bacterial inhibitors of receiving waters. We have not
been able to correlate these factors with the actual
bacteriological, data nor determine how these factors relate
to the differences in coliform/fecal ratios. Tlie Division
of Shellfish Sanitation concluded that any new indicator
criteria should be evaluated according to the sanitary
significance of the specific indicator and not according
to the relationship between the indicator group being evaluated
and tiie present criteria. In review of the fecal coliform
water quality data obtained from State shellfish control
programs, very few of these data had been correlated with
known pollution sources. It has been very difficult to make
concise public health appraisal of these data for lack of
in depth sanitary survey information.
During the Microbiology Task Force deliberations at the
Seventh National Workshop in October 1971, median fecnl coliform
• values o (: 7.8 and
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3
A spokesman from Louisiana stated that a 23 fecal coliform
MPN median would close a high percent of "approved" shellfish-
growing areas in his State which could not be shown to be
affected by known sources of pollution. Delegates from
Rhode Island proposed that for the present coliform standard
that "not more than 10 percent of the samples ordinarily
exceed an MPN of 230 per 100 ml." be omitted from the present
coliform standard. These actions may give you some indication
as to the diversity of opinions expressed in this task force.
Certainly, one must conclude that there is a iNational n'eed
to reconsider and evaluate bacteriological criteria for
approved shel]fish-growing areas. After considerable
discussion, all but two State representatives present in the
task force voted to support FDA's proposal to retain the
\
present growing area standards based upon the coliform indicator
group.
On February 17, 1972, a representative of the Division of
Shellfish Sanitation participated in an Ad Hoc Committee
meeting on bacteriology, which included representatives of
the Environmental Protection Agency (EPA), Oregon Dept. of
Environmental Quality, the academic community, the medical
profession, the National Council on Stream and Air Improvement,
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4
and the pulp and paper mill industry. The significance of
t-
Klcbsiclla pneumonia was discussed. This organism, a member
of the coliform and in part the fecal coliform groups was
reported found in high numbers in paper mill wastes. It
may also be found in other waste effluents high in reducing
sugars and other carbohydrates such as the sugar industry,
food-processing plants, and some domestic effluents. IMViC
reactions will not separate Klebsiella pneumoniae from
Entero_b_acter aerogcnes.
In attempting to interpret existing data on Mobile Bay
waters, there is no way to determine the percentage of the
reported coliform or fecal coliform valuesi that are actually
K. pneumoniae. Such information would be valuable to the
agency responsible for the classification of shellfish-growing
areas in the interpretation of sanitary survey data considering
that EFA has reported that 51% of the total coliform waste
source contribution to Mobile Bay is from paper mill wastes.
During the Ad Hoc Committee meeting, a number of
characteristics of this organism were discussed which are
significant to agencies using coliform or fecal coliform
criteria in the classification of environmental water quality.
These are listed as follows:
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5
a. K. pneumoniae is a member of the coliform group.
b. A large percentage ('10-60S) of the K. pneumoniae
isolated from environmental samples have been reported to
ferment lactose at 44.5°C., and, therefore, vould be reported
in the indicator system as a fecal coliform type.
c. K. pneumoniae has the same IMViC reactions as
Enterobacter aerogenes.
d. K. pneumoniae is heavily encapsulated and," has
f
a longer survival potential in environmental waters than
enteric pathogens or E. co1i.
e. K_. pneumoniae can "colonize" environmental
waters rich in carbohydrates and reducing sugars (such as
paper mil] wastes in Mobile Bay).
f. K. pneumoniae is a normal inhabitant of the
intestinal tract of warmblooded animals, but is generally
found in low numbers compared to F.. co 1 i populations .
g. K. pneumoniae is highly resistant to antibiotics
and is regarded as a "secondary" pathogen by the medical
profession. It usually appears as a pathogen following
treatment of other enteric bacterial infections after normal
populations of bacterial flora have been modified by
antibiotic treatment.
-45-
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6
h. The presence of F.. co 1 i in the intestinal
tract of healthy individuals may restrict the growth of
K. pneumoniac and help prevent "colonization" of the
individual.
i. The presence of effluents enriched with
carbohydrates such as paper mill wastes in shellfish harvest
areas can have an adverse affect on the classification of
such areas, especially if aftergrowth or "colonization".
occurs.
j. Aftergrowth of K. pneumonias can result in
high counts of both coliform and focal coliform groups
which may be totally unrelated to actual pollution sources,
representing real health hazards to consumers of shellfish.
In view of the confusion surrounding an "acceptable"
value of fecal coliform organisms to be used as a substitute
for the present coliform standard, FDA proposed to the
Seventh National Workshop that NS>SP take a closer look at
the more specific 45.5°C test for E. coli. This was first
proposed to the American Public Health Association Committee
on shellfish methodology by its chairman, Dr. C. A. Perry,
in 1936.. Gcldrcich states that the fecal coliform test
at 4-1.5 (i-0.5°)C represents a, ". . . compromise between
acceptable sensitivity and specificity for the fecal types."
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7
For regulatory purposes, the FDA has been successful in the
courts in using E. coli as an indicator of fecal pollution.
Perhaps, the time has come in the history of the NSSP, that
we must re-evaluate our concepts and be willing to lose
sensitivity for an increase in specificity in the promulgation
of both bacteriological criteria and standards for "approved"
shellfish harvest areas. Currently, neither the FDA nor the
NSSP has been convinced of the wisdom of substituting the
* /
narrow spectrum fecal coliform test for the broad spectrum
coliforin group.
If we make errors in standard setting, let it be on
the side of safety. On the other hand, FD.A'and cooperating
State agencies responsible for growing area control have
no desire to prevent utilization of a natural resource by
closing harvest areas where indicator levels exceed present
standards, unless these indicator levels, according to the
best professional knowledge and experience, constitute a
real health hazard.
In conclusion, we realize the need for a more specific
indicator and more realistic standards for use as an adjunct
to the sanitary survey in defining "approved" growing areas.
Any substitute fnr the present criteria should: (1) be more
specif.ic for fecal pollution than the current indicator group,
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8
(2) stand on its own merit as an indicator of actual sources
of fecal pollution without a dependency upon a specific
relationship with the coliform group, (3) be more representative
of the potential pathogen load, (4) consistently indicate the
sanitary quality of shellfish-growing waters in diverse
geographical locations, (5) and have a survival time ratio
in the estuary consistent with that of other bacterial pathogens
regardless of nutrient value of the water.
As previously stated, the Seventh National Workshop, the
V-
f \
NSSP supported the FDA position by a large majority that the
fecal coliform has not adequately fulfilled the qualifications
necessary for the promulgation of a fecal coliform standard
i
for "approved" shellfish harvest areas. The need for a re-
evaluation and renewed effort to develop new criteria is
recognized.
We recommend that 1-. coli be considered as a promising
bacteriological indicator for classifying shellfish harvest
areas and that necessary research be initiated to develop and/or
evaluate rapid tests for the recovery of this organism. The
long-term goal would be new bacteriological criteria and
.standards- for the classification of growing areas. It is
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our considered opinion that this effort should receive
immediate attention and research investment by l-'cderal and
State agencies concerned with shellfish sanitation control,
March 22, 1972
D.. A. Hunt
-1*9-
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- March 24, 1972
COPY
Economic Loss of Oyster Reef Closure
1971 - 1972
This season Alabama's oyster reefs were closed on January 30 and
February 4 (Productive reefs) and have remained closed for the past
50 days.
The average dockside value of the oyster catch is $1,671 or a total
loss of $83,550. However, production this yearvas above average
and was worth $1,916 per day in December 1971. At this figure the
total dockside loss has been $95,800 during the current season.
The dockside value represents only loss to the fisherman. It
has been shown that the total economic value of oyster production
in Alabama is four times the dockside value. Thus, the total
economic loss to the fisheries community as a result of the current
pollution closure is $382,400.
Prepared by the Alabama Department of Conservation and Natural
Resources for presentation at Mobile Bay Technical Committee Meet1"
ing, Dauphin island, Alabama, March 24, 1972.
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Donald W. Lear, 1962, Growth of bacteria in estuarine waters. Chesa-
peake Science 3 (3): 160-165-
—Organisms used were Escher ichia col i, E_. freundi i , and Aerobacter
aerogenes.
—Nutrients were added to filtered distilled water and to filtered
estuarine water (1^.1 ppt).
--Growth was better in the saline waters than in the distilled
waters.
--Cites papers by Kelly and Arcise (195*0 which show increase in
coliform MPN in oysters shipped--indicat ing growth in saline
conditions utilizing food sources within the oyster.
--Implication is that if nutrient sources are available in saline
waters total coliform count will increase.
Gallagher, T.P. and D.F. Spino, 1968, The significance of numbers
of coliform bacteria as an indicator of enteric pathogens.
Water Research, Vol. 2, 169-175.
The traditional concept of using levels of total coliform
densities to describe bacteriological acceptability has been ques-
tioned by many because coliform may result from causes other than
fecal pollution. Because fecal coliform densities result from
pollution by man and other warm blooded animals, they are more
directly indicative of the probable presence of the enteric
associated pathogens. However, summarized data from several
stream surveys over the past few years show little apparent
correlation between quantities of total or fecal coliform and
the probable isolation of salmonellae. Salmonellae were isolated
at total coliform densities of less than 1000/100 ml and fecal
coliform densities of less than 150/100 ml. The total coliform
density of 1000/100 ml has traditionally be«_.i acceptable for recrea-
tional waters. It is probable that salmonellae will be isolated
at high fecal coliform densities but observed data has indicated that
isolation is also probable at low coliform densities if there is an
Prepared by the Alabama Department of Conservation and Natural
Resources for presentation at Mobile Bay Technical Committee Meeting
Dauphin Island, Alabama, March 2*1, 1972.
-------
inadequately disinfected v/aste source which may affect the area of
concern. In one instance cited, the area was 73 miles and 4 days
travel time downstream from sources of waste. This may be because
salmonellae are persistent under conditions which may be adverse
to survival of fecal coliform. Large percentage reductions of
coliform without chlorination will not of itself guarantee bacter-
iological acceptability. Data show that salmonellae have been
isolated after a 99 percent reduction in fecal coliform below unchlorinated
waste sources.
Although salmonellae at present are the only pathogens which
may be isolated from water by means of a routine test with any
degree of reliability, it is probable that methods for other
pathogens will be perfected in the near future. Until that time,
the coliform will still be valuable as indicators of pollution.
However, low values of total and fecal coliform are not, of them-
selves, sufficient to indicate bacteriological safety from patho-
genic organisms. A control program to more effectively evaluate
bacteriological safety in recreational waters should include: (l) a
sanitary survey to delineate possible sources of pollution;
(2) effective disinfection of all waste sources which may affect the
area; (3) use of the salmonella test in addition to total and fecal
coliform as a monitoring device.
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Rubin, A. J., et al, 19&9, Coagulation of Escherichia coli by neutral
salts. Water Research. Vol. 3, 8^3-852.
Shows £. col i to coagulate at about 23 ppt NaCl (.^tlSN) at
a pH of 6. Which is beyond the desirable salinity range for oyster
production and indicates only dilution reduces the number of
coliforms. This dilution is made by decrease in freshwater flow
in relation to saline waters within the bay. pH's in the range
of the bay water cause no increase in aggregation or coagulation.
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ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF ENFORCEMENT
DIVISION OF FIELD INVESTIGATIONS-DENVER CENTER
BUILDING 22 , ROOM 410 , DENVER FEDERAL CENTER
DENVER, COLORADO 80225
TO .- Files DATE: 29 March 1972
FROM : Robert F. Schneider
Chief, Biology Branch
SUBJECT: Mobile Bay Technical Committee Meeting, Dauphin Island, Alabama
March 24, 1972.
Attendees of the March 24, 1972 Technical Committee Meeting to
consider Recommendation No. 6 of the Mobile Bay Enforcement
Conference were:
Committee members: Representing
W. F. Anderson Alabama Department of Conservation and
Natural Resources
James Coil, Jr. Scott Paper Company, Mobile, Alabama
Arthur Beck Alabama Water Improvement Commission
J. David Clem FDA, Washington office
C. B. Kelly EPA, Washington office
Robert Schneider Chairman
F. J. Silva (absent EPA, Atlanta office
because of illness)
Others:
Albert Story FDA, Dauphin Island, Alabama
Eddie May Alabama Marine Resources, Dauphin Island,
Alabama
Wayne Swigle Alabama Marine Resources, Dauphin Island,
Alabama
Committee member spokesmen for the State of Alabama discussed the
present status of shellfish harvesting areas in Mobile Bay. They
informed the Committee that the Bay oyster production in December,
1971 was nearly $300.00 per day higher than it had been for the
past several years. On February 4, 1972 all productive reefs in
Mobile Bay were closed and have remained closed for the past 50
days. Closure has resulted in an economic loss (total dockside)
of $95,800. The dockside value represents only loss to the fisherman.
Alabama State Officials explained that the total economic value
of oyster production in Alabama is four times the dockside value;
thus, the total economic loss to the fisheries community as a
result of the current pollution closure is $382,400.
-------
Page 2
Memo to Files
Re: Mobile Bay Technical Committee Meeting
March 29, 1972
Discussions during the remainder of the meeting dealt with the
various bacteriological criteria that might be substituted for the
total coliform criterion presently used in evaluating shellfish
harvesting waters. The fecal coliform indicator group was the
principal substitute criterion considered. The FDA spokesman
told the Committee that neither the FDA nor the MSSP is convinced
of the wisdom of substituting the narrow spectrum fecal coliform
test for the broad spectrum coliform group. Contrasting views
were expressed by other attendees. They pointed out to the
Committee that fecal coliform densities result from pollution by
man or other warm blooded animals; therefore, the fecal coliform
test was more directly indicative of the probable presence of enteric
associated pathogens than the "broad spectrum" coliform group.
Although the Committee did not reach an agreement (during this session)
on a more specific indicator or more realistic standards for Mobile
Bay, we did establish the qualifications any substitute criterion
must have. Based upon a prepared statement by the FDA representative,
the Committee agreed that the substitute for the present criterion
should: (1) be more specific for fecal pollution than the current
indicator group, (2) stand on its own merit as an indicator of
actual sources of fecal pollution without a dependency upon a
specific relationship with the coliform group, (3) be more represen-
tative of the potential pathogen load, (4) consistently indicate
the sanitary quality of shellfish-growing x%Taters in diverse
geographical locations, and (5) have a survival time ratio in the
estuary consistent with that of other bacterial pathogens regardless
of nutrient value of the water.
Within the Committee, a working committee was established. Committee-
men Clem and Kelly agreed to jointly prepare a statement suggesting
an approach to assist regulatory agencies in the development of a
meaningful bacteriological criterion for Mobile Bay shellfish harvest-
ing waters. The statement will be submitted to the Committee Chairman
by April 14, 1972 and subsequently distributed by the Chairman to
Committee members for their review and comment.
On May 5, 1972 the Technical Committee will meet at Dauphin Island,
Alabama to discuss the statement prepared by the working committee
and to deliberate a relevant recommendation to the Conferees.
Robert F. Schneider
RFS/ls
-55-
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APPENDIX D
CORRESPONDENCE CONCERNING A PROPOSED RECOMMENDATION
-56-
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ENVIRONMENTAL PROTECTION AGEN^V
OFFICE OF ENFORCEMENT
DIVISION OF FIELD INVESTIGATIONS- DENVER CENTER
BUILDING 22, ROOM /»10 , DENVER FEDERAL CENTER
DENVER, COLORADO 80225
TO = DAiE:2A April 1972
Mobile Bay Technical Committee Member
FROM : Robert F. Schneider, Chairman
SUBJECT: Technical Committee to Consider Recommendation No. 6 of the Mobile
Bay Enforcement Conference.
Following our deliberations in Dauphin Island, I have had the oppor-
tunity to talk with some of you by telephone concerning the Mobile
Bay shellfish harvesting problem. From these conversations and from
the materials presented to the Committee during our March 2*», 1972
meeting I have developed the opinions contained herein.
There is a Committee consensus that a substitute criterion for the
total coliform test is needed for evaluating shellfish harvest
areas in Mobile Bay. Those who are responsible' for criteria develop-
ment (i.e. FDA shellfish Sanitation Branch and EPA Office of Water
Programs) should consider the following characteristics in the design
of a substitute bacteriological criterion. The "new" bacterial indi-
cator should: (1) be more specific for fecal pollution than the
current indicator group, (2) stand on its own merit as an indicator of
actual sources of fecal pollution without a dependency upon a specific
relationship with the coliform group, (3) be more representative
of the potential pathogen load, (*») consistently indicate the sanitary
quality of shellfish-growing waters in diverse geographical locations,
(5) and have a survival time ratio in the estuary consistent with that
of other bacterial pathogens regardless of nutrient value of the water.
The Committee considered two alternative indicators as the most promising
for evaluating bacteriological contamination in Mobile Bay shellfish
harvest areas, (l) fecal coliform or (2) Escherichia coli.
Committee members and other attendees at the March 2k Dauphin Island
meeting pointed out certain benefits and weaknesses of these substitute
indicators. The major points were:
Presently, the fecal coliform test requires less time and effort than
the E_. col i measurement; therefore, results can be obtained sooner,
at less expense, and by fewer microbiologists. However, studies by
the Gulf Coast Laboratory and others have failed to demonstrate a
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Page 2
Memo to Mobile Bay Tech Comm Mbrs
2*» April 72
consistent total coliform/fecal coliform ratio or arrive at a fecal
coliform value that can be directly related to known sources of
fecal pollution and provide public health protection to consumers
of raw shel1fi sh.
Sanitary surveys and published documentation confirm the fact that
the presence of E_. col? consistently indicates fecal pollution and
the potential contamination by other pathogenic bacteria.
Based upon these considerations and in compliance with Recommenda-
tion No. 6 of the Mobile Bay Enforcement Conference, the Technical
Committee submits the following statements to the Conferees: "To
assist in the development of a more realistic criterion than the
present method for evaluating the probability of pathogen presence
in Mobile Bay, it is recommended that the criterion be based on the
measurement of Escherichia coli in waters overlying shellfish harvest
areas. Furthermore, the Technical Committee recommends that the
Federal and State shellfish sanitation control agencies combine their
efforts to support the research needed to develop a rapid, reliable
method for enumerating E_. col i in estuary waters used for shellfish
harvesting."
At present, t suggest we postpone the tentative meeting scheduled at
Dauphin Island on May 5i 1972. I would like to give each of you the
opportunity to respond to the recommendation contained herein. If
possible, I would like your written comments by May 10, 1972. If
you feel further Technical Committee meetings would be fruitful, please
advise me and I will set up a mutual meeting place and time.
Robert F. Schneider
RFS/ls
cc: Mr. John White, Atlanta, GA
Mr. Murray Stein, D.C.
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STATE OF ALABAMA
** DEPARTMENT OF CONSERVATION AND NATURAL RESOURCES
f, 0. Boi 188 - Dauphin Island, Alabama 5652S
G. !GE C. WALLACE
GOVERNOR
'• '.UDE P. KELIEY
3MMISSIONER
: 1JEY D. BLtDSOE
SISTANT COMMISSIONER
April 28, 1972
^
-*1 -V'.T
.- ..v 1-. ' &\
* W.\% -
•-.. W
'O. :-,-> CV/
DIVISION OF MARINE RESOUR
WIUIAM f. ANDERSON. DIRECTC
Mr. Robert F. Schneider, Chairman
Mobile Bay Technical Committee
Environmental Protection Agency
Office of Enforcement
Division of Field Investigations
Building 22, Room 1±10
Denver Federal Center
Denver, Colorado 80225
Dear Mr. Schneider:
We concur with your opinions 100 percent. Originally ire felt that fecal
coliform would be a better standard. However, after learning that Klebsiella
is detected in the test for fecal coliform and that it also is frequently
associated with paper mill effluent, we certainly agree that E. coli would be
a much better indicator organism. We feel this way because the discharge of
paper mill effluent at the head of Mobile Bay has a population equivalent in
excess of 1*00,000.
It was suggested by some of the FDA research group here on the island that
a salinity level of 10 ppt be used as the closure point. However, on examination
of some of the data compiled by them (attached) there seems to be little corre-
lation between this level and high coliform levels. Also their supposition is
still based on the 70 MPH/10 cc of total coliform in relation to salinity. We,
therefore, feel that the criterion you have suggested would be much better.
We are sending under a separate cover some of our publications on Mobile
Bay which might be helpful to you for background information.
Sincerely,
W. F. Anderson, Director
Marine Resources Division
WESrjb
cc: Mr. Claude Kel3ey/w copy of Mr. Schneiders letter
Mr. Arthur N. Beck
-59-
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22
9 May 1?72
Mr. W, F. Anderson
Marine Resources
Dcpartnont of Conservation
and Natural Resources
P.O. Box 135
Dauphin Island, Alabama 3^523
Dear Mr. Anderson:
Thank you for your prompt reply to my letter of April 2*:, 1972.
I have not received opinions fron all the Corr.ii tts?. ncnbers
yet, but the responses fron FDA rand Scott Paper arc in agreement
with the "E. ceil? indicator'1 recommendation. V/hen ! receive the
others I wTl 1 draft a final recon-.nendation and send it to you-
alonn with the letters I've received from the Coraiittee for your
review and comment.
Aijaln, thank you for your help and for sending no the series of
Alabama Marine Bulletins and oyster resource atlas.
Sincerely,
Robert F. Schneider
Chief, Clolooy Branch
RFS/Is
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TMENT OF HEALTH. EDUCATION, AND WELFARE
PUBLIC HEALTH SERVICE
FOOD AND DRUG ADMINISTRATION
WASHINGTON. D C 20204
May 5, 1972
Mr. Robert F. Schneider
Chairman
Mobile Bay Technical Committee
Room 410, Building 22
Denver Federal Center
Denver, Colorado 80225
Dear Bob:
I concur with your letter of April 24 but would like to
make the observation that one study conducted by our
staff indicates that E. coli can be recovered within
24 hours with results comparable to present methods.
Dr. Wallace H. Andrews' paper on this study is attached
and has also been published in Applied Microbiology,
Vol. 23, No. 3, March 1972 issue. Of course, their
work needs further collaboration but it does look
promising.
If I can be of any further assistance, please call.
Since/ely ,
J. David Clem
Director
Division of Shellfish Sanitation
BF-230
Office of Food Sanitation
Bureau of Foods
Enclosure
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SCOTT PAPER COMPANY
SOUTHERN OPERATIONS
P. O. BOX 2447
MOBILE, ALABAMA 36601
JAMES H COIL. JR.
MANAGER OP
AOMINIS1 H At 1 VC SERVICFS
May 5, 1972
Mr. Robert F. Schneider
Environmental Protection Agency
Office of Enforcement
Division of Field Investigations-Denver Center
Building 22, Room 410, Denver Federal Center
Denver, Colorado 80225
Dear Mr. Schneider:
Subject: Technical Committee to Consider Recommendation
No. 6 of the Mobile Bay Enforcement Conference.
I would suggest consideration of altering your statement to the conferees to
read as follows:
"To assist in the development of a more realistic criterion
than the present method for evaHiating the probability of pathogen
presence in Mobile Bay, it is recommended that the criterion be
based on the measurement of Escherichia coll in waters overlying
shellfish harvest areas. The Technical Committee further
recommends that the agencies concerned, i. c, , FDA Shellfish
Sanitation Branch, EPA Office of Water Programs and the Alabama
State Department of Health, initiate as soon as possible stirveys of
the E. coli counts—in addition to total coliform—in the waters overlying
the shellfish areas and on the shellfish proper. Furthermore, the
Technical Committee recommends that these aforementioned federal
and state shellfish sanitation control agencies combine their efforts
to support the research needed to develop a rapid, reliable method
for enumerating E. coli in estuary waters xised for shellfish harvesting. "
t Very truly yours,
' ' •'••'/
/' James H. Coil, Jr."
Paper Industry Representative
,' /Mobile Bay Technical Committee
JHCtgor
cc: Mr. John White, Atlanta, GA
Mr. Murray Stein, D. C. ,
-------
22 MO
11 Hay 1972
Mr. James Coil
Manager of Administrative Services
Scott Paper Company
P.O. Eox 2^7
Mobile, Alabama 3&&01
Dear Mr. Coil:
To date I have received replies frcn the State, FDA and you con-
cernlnu the recoitiT.erKbtion ! prepared In sr.y letter of April 2J»,
1972. Generally, these Cor.inilttec members concur with the "E_. coH_
Indicator" aoproach. When I receive comments frora the entire
CoiTiKittec t v;ill amend the recor.n?endatlon, prepare ii final clraft
and nail a copy to you For cerements.
May I take this opportunity to thank you for your help on this
technical coraalttee.
Sincerely,
Robert F'. Schneider
Chief, Biology Branch
RFS/l!
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ENVIRONMENTAL PROTECTION AGENCY
Office of Water Programs
Washington, D.C. 20460
May 15, 1972
Mr. Robert F. Schneider
Office of Enforcement
Division of Field Investigations
Building 22, Room 410
Denver Federal Center
Denver, Colorado 80225
Dear I'ir. Schneider:
This is in response to you<" letter of 24 April requesting
comments on a proposal to recommend that Federal and State
Agencies combine their efforts to support the research needed
to develop a rapid, reliable method for enumerating E. coli
in estuary waters used for shellfish harvesting. On this
issue, I have the following comments:
1. I have the sincere opinion that the Technical
Committee is not in a position to recommend the choice of any
one indicator system.
2. The matter of choice of indicator system properly
should be made only after careful study of the available data
and further cu*ta gathered in future investigations.
3. In my opinion, further study should not be conducted
only on E. coll. To be meaningful, other coliforms, including
total coliforms and fecal coliforms should be included in the
investigations. Including these others would provide a bridge
from past experience and data on the other organisms and, if
contraindication for E. coli are found, more supporting
information for coliforms or fecal coliforms would become
available.
7
C. B. Kelly
Special Assistant for
Criteria and Standards
Water Supply Programs Division
-------
EN'"RONMENTAL PROTECTION AG[ :Y
Program Grants Section DATE: May 2U, 1972
SUBJECT
TO: 'Mr7Itobert F. Schneider
Chief, Biology Branch
Denver Center
From your memorandum and attached paper by Clem and Hunt, I
presume that the direction for seeking a suitable indicator organism .
to aid in the sanitary classification of shellfish grov/ing areas lies
toward the utilization of the organism, E. Coli.
The course which is recommended represents a progressive step
toward our refining the bacteriological spectrum of the general coliform
organism group for use in sanitary classification of shellfish growing
areas . First, the coliform test was used for classification purposes,
then in the early 1960's - and after a lot of research - the "fecal
coliform" test was accepted as a supplementary test for classifications.
The test, which was widely used to obtain supplementary information, was
never accepted as an alternate to the coliform test. Now going to the
E. Coli test represents a narrowing of the spectrum.
The proposal in your memorandum suggests that a stxidy be made to
determine the suitability of using E. Coli as an indicator organism
This may be a good realistic approach to establish an organism with
fecality integrity which will provide a true index of the sanitary quality
of shellfish growing waters. There has been, however, a great deal of
experience utilizing the fecal coliform group in shellfish work and a
lot of research too. First, I suggest that the committee not be hasty
in turning away from this accumulated experience and, therefore, recommend
that a hard look be taken at the vast amount of F.C. information which
has been collected and applied. Secondly, if the committee elects to
move forward toward encouraging the development of a rapid and accurate
E. Coli test, then such development must move forward with all deliberate
speed.
It would be interesting to have C. B. Kelly's and Dr. Hosty's
comments on the suggested course of action before a firm commitment to
move ahead with the E. Coli study is made. It would also be interesting
to look into the standards and procedures used by the Eglish, French,
-65-
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Portuguese, Spaniards, and Japanese in utilizing bacterial indices for
classifying shellfish growing areas. While it may not be necessary, I
hope we don't get involved in a protracted study - which includes that
more study is needed before definite recommendations can be put forth.
J. Silva
-66-
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State of Alabama
Department of Public Health
State Office Building
Montgomery, Alabama
36104
IRA L. MYERS, M. D.
STATE HEALTH OFFICER
May 22, 1972
Mr. Robert F. Schneider
Chief, Biology Branch
Environmental Protection Ag.-ncy
Office of Enforcement
Bldg. 22, Room 410, Denver Federal Center
Denver, Colorado 80225
Dear Mr. Schneider:
to o- z have asked our Director of Laboratories, Dr. Thomas S. Hosty
to give me hj.s ideas and recommendations in regard to proposals to evaluate
measurements of pollution in Mobile Cay harvesting areas evaluate
Yours^yery truly, _
"'"
AMB/.
f rthur tf. Beck
Chief Engineer & Director
Bureau of Environmental Health
cc: Dr. Thomas S. Hosty
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IRA L. MYERS. M. D.
STM E HEALTH OFFICER
State of Alabama
Department of Public Health
State Office Building
Montgomery, Alabama ism
August 7, 1972
lir. Murrny Stein
Chief Enforcement Officer - Tteter
Crystal Hall Dldg. 2
KoOal lilG
U&Bhington, B. C. 20460
Dear Iftr. Stein:
I apologize for not edvleing I.'r. Robert F. Schneider, Chairman,
llobllc !5ny Trtchxiical Cj-.ui.lt toe before now of r™ srecoa^cndRtion In regard to
an indicator tost ;:."»r pollution control. oi: oystc-r j;ra:;inj; waters.
ProbcbLy wore work hcs bcon done 0:1 labile 2=y waters with retard
to oyscor 2>:o-.-:i«2 v:;t.crs th:
-------
Mr. Murray Stein
August 7, 1972
-2-
Schr.eider.
Again I apologize for being late with my report to you and Mr.
ANB/iw
end.
Yours very truly,
mr M. Beck
Chief Engineer f* Director
Bureau of Environmental Health
cc: Mr. Robert F. Schneider
Dr. Thomas S. Kooty
-69-
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State of Alsbssna
.'. t*-, „. r. .J. —A .}•>•.lm ».?i'
ImJiU 0) /:.,,•,;b ji'
Bi!rc?u of l.r.!;oni'jriHs
434 Monroa Strest
Hontgonisry, Aiaisma 3G1C4
IRA L. MYEPS. M.
STATE HF.ALTH
MEMORANDUM
TO:
FR011:
SUBJECT:
August 4, 1972
Mr. Arthur Beck
Thomas S. Hosty, Ph.D., Director
Bureau of Laboratories
Mobile Bay
THOMAS S. HCSTY. PH.D.
DIRECTOR LABORATORIES
I did not write to Mr. Schneider as I tolcl you I would since I felt
that the meeting we had in Virginia would yivc a satisfactory explanation
of Alabama's stand. All of the testimony given in Virginia was transcribed
and should be readily available to FDA. Many of the representatives
of FDA were present at this meeting. I think it is worthy of note chat the
opposition of the FC cest was soru or taKcn apart very nicely. I was
extrofuely interested Ln the facu thct psrhans two of the nost knowledgeable
people in the country on cvstcrs and seawater were in co-./iplete agreement;
that the fecal col it'ovn tcse should be u:,£d rath or than coliLor-.Ti. L sm, of
that every ncetin^ I 50 to I ani ccid there is not sufficient data co justify
focal col i forms. Wa ware advised before t;oip.:; to Virginia that we should
be able, to document w'.at we were savins. Through the years the opposition
remains tree to say any;ning they please - mostly without adequate
documentation.
Recent developments in this area have left me completely confusod.
Certainly we are not rgainst tiie development of a better test. 1 .r.«cle
a statcnent in Virginia chat '.-.ion they speak cf "E.coli'1 test "hat do
they wean:' 1 stated that none or the tests we use can be considered
as an E.coli test unless yoj IMViC the strains and no one was in
opposition, to this sentiment. Mr. Kelly very beautifully folLoved
the history of th-? feccu coliform test and pointed out that even the
author of the E.coli lost did not consider this to be a specific test
for tt.coli.
£'
A considerable amornt of discussion li.is been made relative to the ,-"
growing of E.acrR»rc nos in aifluence from paper mills. Certainly it is '.'
proba'uly lorjic.il to assume that with tiic waste products chat are present '•
and ;i tc-.iiporaturc c:i iiro-mci 32" tha:. growth wouJd tr,
-------
Stats of A
y rv? •.iiiV.j-
., OS i .i...i
Bureau of Ui'orator.
434 Monroo Sinst
f.l&nlscir.sry, Aiibuiia J
IRA L. MYCRS. M. 0.
STATC HEALTH OFFICER
TO: Mr. Arthur Beck
THOMAS S. HOSTY. PH.D.
DIRECTOR LABORATORIES
August 4, 1972
Page 2
and subsequent tests. It was found later that: sewerage from the plant
was discharged into paper mill waste with the F.C. responding very nicely.
Mr. Salinger from Maryland presented a very nice study on the
relationship of fecal coliform results to polluted, clean and intermediate
typos of pollution and that study has become part of the record of the
Virginia meeting. A copy of Mr. Salinger's report is included with this
memo as well as the study on the new Scott Paper mill.
I am also enclosing a part of our study of station 119 and it shows
rather dramatically not only the ability cf the fo.c^l coliform test to
detect pollution but coinc Ldcs quite well with the dropping of salinity.
These studies were duplicated on at least 25 other stations during the
course ol three years.
May 1 refer you to the Denver, Colorado reefing of Xay 10, 1972
with microbiologisus of iiPA unu one of uhi:ir co7iciuijj.c-.ir,: "The fecal
coliform is a valid indicator of fecal" pollution in ail waters."
I was very interested in the continuous referral of a test
developed by the Uauvjnin Island workers which was entitled
"The Rapid Recovery of E.coli fror> Estuarine i.'aters."
The article appears to have considerable premise but rcaliv my qood
friend Mr. Prcsncli •..••xiLd agree that the only difference between the
E.coli and his test va.*. a matter of 24 hours or a quicker test:. We
are at present doing a limited study of the tost on oysters but a^ain
T aii certain Mr. Pres^oil would not advoc.itc thar tnis test be adopted
immediately without confirming tests on n much wider area since the area
where these studies wore made is not generally of tne same character as
the areas over th*. ovsccr beds. At any rate we will continue our limited
studies as time and help will allow.
For many years the cooperative agreement between states and shellfish
people has benefited the whole shellfish program and may I ask now is
this fifty years of cooperative agreement to be nullified by FDA calling
the fhots.' Our data ovf.-r three years showed the FC test could be used in
Alabama with a cut of! MPN of about 9.0. This JlFN would still -ivc .adequate
warning OL possible unLiution and yet would allow the beds to be reopened
sometimes even four to live weeks before the col i torv.is returned to normal.
Certainly our firr.t ou! :".ation r.iiouu! be the neaicli urobLcm buL ;it the same
tima wo should bn a-./.-u-1.- uiat n-nnv pconic arc nur. out of work i r t!;c indicator
is «'i ; '.o'.i i u-.-'ii:; i L i \ i- r-.-iuiiro i.n.'ii, it ; i'-nai-j r-»si iiicion VIMMI r.iu-ro ronllv
i.. • :'.-'. ii !i!.l'.!:! L'.I.: .:<.: .'"..'li av.lli.ii)lo \-;.- vi : i ! ,,..!,»•.• ..Ill 1. • 11.1.' n.-. 1 '. C1 ••'.••.I'.y
again OL the uxmciu;1. LOJC and aLLcr.ipt to dcvciup new
-71-'
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APPENDIX E
RAPID RECOVERY OF ESCHERICHIA COLI
FROM ESTUARINE WATERS
-72-
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COPY
ABSTRACTS OF RAPID METHODS FOR THE RECOVERY OF Escherichia coli
Three rapid methods for the recovery of Escherichia colis which
appear applicable to shellfish and shellfish waters, are abstracted.
METHOD A
Fishbein et al. investigating the recovery of E. coli from
marine products and other foods found that the best recovery was
obtained from lauryl sulfate tryptose (1ST) broth incubated at kb C.
The method is a fermentation tube technic in which the samples are
inoculated directly into LST broth and incubated at bk C in a water
bath. Since 96 percent of the E. coli recovered by the method
were recovered with 2k hours it is essentially a 2^-hour test.
Results obtained by the method were comparable to those obtained by
the Association of Official Analytical Chemists (AOAC) test. The
latter test is much longer, requiring from two to four days.
A complete discussion of the method appears in Applied Microbiology3
Vol. 15, No. 2, pp 233-238, under the title, "Coliform Behavior in
Frozen Foods."
METHOD B
Andrews and Presnel1 of the Food and Drug Administration, Gulf
Coast Technical Services Unit, have developed and evaluated a 2A-hour,
elevated temperature, tube fermentation, most probable number test.
The medium, designated as A-l, contains concentrations of lactose,
tryptone, and sodium chloride found in most commercial media. In
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addition the medium contains trition X-100, a surfactant, and salicin,
a carbohydrate readily utilized by E. coli. This method involves the
direct inoculation of the sample, in appropriate dilutions, into the
medium which is incubated for 2k hours at M.5 C in a water bath.
The method has been compared with the procedure contained in the fourth
edition of APHA Recommended Procedures for the Bacteriological
Examination of Sea Water and Shellfish and the 1ST elevated temperature
(M C) test described by Fishbein. The APHA recommended method
requires a 2A-to-48-hour presumptive test at 35 C followed by a
2^-hour confirmatory test at kk.S C. The three tests were compared
using samples of freshwater, estuarine water, and a limited number
of shellfish samples. Results of the comparative study demonstrated
that E. coli recovery by A-l medium at kk.$ C and LST at M C were
comparable and that both methods recovered more E. coli than the APHA
recommended procedure. The report pertaining to the development and
evaluation of the method is included in this Appendix (pp 72 to 82 ).
METHOD C
Cabelli and Heffernan have described a rapid, elevated tempera-
ture plate count method for assaying the E. coli content of seawater
and shellfish. The medium used is MacConkey's agar modified by
decreasing the oile salts concentration from 0.15 percent to 0.075
percent. The sodium chloride eliminated from the medium is replaced,
to varying extents, by the addition of shellfish and seawater samples
mixed with the medium. In using the method, known weights of
-------
shellfish meats or known volumes of seawater are mixed with a known
volume of the medium and distributed into petri plates. The plates
are incubated at ^5-5 i. 0.5 C for 2k hours. E. coli appear as brick
red subsurface colonies. A description of the method is published in
Applied Microbiology, Vol. 19, No. 2, pp 239-2^.
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RAPID RBCOVERY OP Bscherichia coli FROM ESTUARINB WATERS
WALLACE H. AHDREBS
MATOARD W0 PRESHBLL
U.S. DBPARTMBOT 0? HEALTH, EDUCATION, AHD WBLFARB
Public Health Service
Food and Drug Adesiaiotration
Bureau of Foods
Office of Food Sanitation
Division of Shollfiah Sanitation
Gulf Coast Technical Services Unit
Dauphin Island, Alabaaa
-76-
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ABSTRACT
The efflciences of two 24-hr elevated-teaperature tests to
recover E. coll froa estuarine water vere coapared siaultaneously
with the 72-hr standard ncthods procedure of the American Public
Health Association (APHA). Froa 1,710 tubes, E^ cgli waa recovered
222 tiaes in lauryl tryptose aiedium incubated at 44 + 0.2 C for 24
hr, 261 tiaea in an expericantal aediua incubated at 44.5 + 0.2 C
for 24 hr, and 257 times by the 72-hr APHA oethod. The muaber of
false positives enunerated was sieilcr in all three tests. The data
indicated that E. coli in raw seawater could be deteriaincd in 24 hr
without a significant loss of accuracy.
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INTRODUCTION
The need for a rapid, reliable method for determining the bacterial
quality of ohellfish-growlng waters is an important aspect of the con-
tinuing search for methods improvement. The standard methods procedure
for the examination of water recosaended by the /uaerican Public Health
Association (APHA) requires a maximum 48-hr enrichment period in lac-
tose broth or lauryl sulfate tryptose broth (LST) to obtain optimum
recovery of fecal coliforas. Positive tubes are then transferred to
EC Tsedium to determine the most probable number (KPN) of fecal coli-
foras. Gas production with 24 hr in the fomentation vial of a tube
of EC medium incubated in a water bath at 44.5 +0.2 C is considered
a positive reaction Indicating an organism of fecal origin. Tubes
that do not produce goo indicate that that the organisms are derived from
sources other than the tract of a warm-blooded animal (1). Because of
the reported interference by non-fecal colifore biotypes, notably
Ettterobacter oerogenen and Eachorichle internediua types, with the
specificity of the EC mediua even at teaiperatures of 44 C (7,8) and
the APHA-adopted 44.5 ± 0.2 C (3,4,6), it is suggested that a determina-
tion of the E. coll density would give a wore accurate estimate of
the degree of fecal pollution of a water sample. Working with frozen
foods, Fishbein et al. (5) reported that LST incubated directly at 44 C
gave E. coli recovery similar to that of the standard methods procedure
of the APHA. A preliminary investigation in our laboratory demonstrated
that a newly-formulated medium also gave E. coli recovery comparable to
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that of the APHA tsethod. The purpose of this study was to compare,
simultaneously, the reliability of E. coll recovery from field oeawater
sasples by use of (i) Fishbein'a elevated-tceperature test, (ii) an
experimental 24-hr elevated-tecperature teat with the newly formulated
median, and (iii) the 72-hr APHA standard sethods procedure.
MATERIALS AND METHODS
Sixty-eight saeples of seavater froa various sites in Mobile Bay were
collected, iced, end transported to the laboratory. All sacples were
tested within 24 hr. after collection. Five-tube decimal dilutions
were perforead in phosphate buffered water (pH 7.2) ea far as necessary
to obtain a determinant KPN of colifores. Test IA (LSI incubated at
35 + 0.5 C in a walk-in Incubator for 24 to 48 hr) end Tect IB (positive
tubes froa Test IA transferred to EC cediusa and incubated in a water
bath at 44.5 j; 0.2 C for 24 hr) together comprise tha APHA standard
methods procedure for the examination of water (1). In Test II, LST,
and in Test III, Kediua A-l (the newly forculated ctediun) were inoculated
and incubated directly in water baths at 44 ± 0.2 C and 44.5 + 0.2 C,
respectively. Medium A-l included 5 sleecoce 20 g tryptone, and 5 g NaCl
found in most rf the coasaercially prepated dehydrated media; and, in
addition, 1 ml Triton X-100 (Roha and Haas Co.)*, a surfactant, and
0.5 g salicin (Difco), a carbohydrate readily utilized by Bacherichia
species (2), added to 1,000 ml of distilled water.
Mention of co*baerlcal products does not iotply endorsement by the
Food and Drug Adainiatration.
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All positive tubes from Teats IB, II, and III were streaked on
Levine's eosin nethylene blue agar and incubated at 35 + 0.5 C for
24 hr. One representative colony of each type was picked to lactose
broth and incubated at 35 C for 24 and 48 hr. All positive tubes
(gas formation) were inoculated into 1% tryptone, M.R.-V.P. medlua,
and Koser citrate medium and subjected to indole, rcethyl red, Voges-
Froskauer, and citrate (IMViC) determinations at 35 + 0.5 C. All
positive lactose broth tubes were also reinoculated into EC medium
and incubated at 44.5 j; 0.2 C fox 24 to 48 hr to confirm the ability
of the pure culture to grot; and produce ges at the elevated tempera-
ture.
To facilitate a comparison with the d&ta of Fishbein et si.(5),
we decided to use their terminology, which in cs follows: positive
tube, a tube that ferments lactose with the production of gas within
24 to 48 hr; negative tube, a tube that does not produce gas within 24
to 48 hr; B. coli , a tube frosa which E. coli Type I (-H- —) or Type
II (-+--) is isolated; false-positive tube (FP), a tube that gasscs
within 24 to 48 hr, but frosn which E. coli+ is not isolated; false pos-
itive (group), a tube that gasses within 24 to 48 hr, but from which a
coliforn organise other than E. coli* is isolated; false positive (syner-
gistic), a tube that gassea within 24 to 48 hr, but from which neither
E. coll* nor any coliforn is isolated.
RESULTS AND DISCUSSION
In Table I, the positive tube reactions of the four tests are compared,
LST incubated at 35 C (Test IA) produced the largest number of positive
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tubes; all the elevated-temperature teats (IB, II, and III) produced
considerably fever positive tubes. LSI incubated directly at 44 C
(Test II) produced the fewest positive tubes (631), whereas EC medium
(TEST IB) and Mediua A-l (Tect III) incubated at 44.5 C produced a
similar mu&ber of positive tubes (665 end 674).
E. coll recovery data ore given In Table 2. Not as nany B. coll
were recovered in Teat II ao in ?G8ts IB and III. The productivity
ratios (coiutsn 3) were obtained by dividing the number of tubes from
which E. coli+ waa isolated by the total nuober of gassing tubes.
The observed differences in productivity ratios of the threo tests
were not considered significant.
Data for the recovery of coliforns; exclusive of E. coll Types I
and II, are given in Table 3. These data indicate no significant
difference in the nusher of faloe-pocltive tubes enumerated by the
three elevated-temperature testa.
The results of this ctudy are consistent with those of Fiohbein
et al. (5) who found that LST incubated at 44 C gave B. colt recovery
comparable to that obtained by the APHA standard icethods procedure.
Although with LST incubated at 44 C we did not recover as many coliforaa
(631 contrasted to 665) and B. coll (222 contrasted to 257) as were
recovered with the standard methods procedure, the differences were not
significant.
We fcioh to emphasize that 44 C, rather than 44.5 C, was used as the
temperature of direct incubation of the LST in the water bath, inaaouch
as this was the temperature used by Fiohbein et al. (5). Furthermore, we
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Table 1. Positive tube reactions
(1)
Test
LST, 35 C, presumptive (IA)
EC, 44.5 C, confirmatory (IB)
LST, 44 C (II)
Kediuta A-l, 44.5 C (III)
(2)
Nusber of
positive tubes
1011
665
631
674
(3)
Percentage of
Cubes positive*
59.12
38.89
36.91
39.62
•=Based on the nuober of pocitive tubes divided by the total number
of tubes (1,710).
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Table 2. Recovery of K. coll
(1)
. Test
(2)
B. coli*
tubes
E. colf+Vtotal
no. of gassing
EC, 44.5 C, confireatory (IB)
LST, 44.C (II)
Medium A-l, 44.5 C (III)
257
222
261
38.65
35.18
38.72
lEjrpriissed as a percentage.
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Table 3. Coliform group recoveries -
(1)
Test
EC, 44.5 C, con-
firmatory (IB)
LST, 44 C (II)
Medium A-l, 44.5
C (III)
-All Enter ohocter
(2)
Total
no. Ppb
406
407
405
(3)
No. FP
(group)
385
389
380
(4)
No. FB
(synergiatic)
21
18
25
and Eecheriebia species exclusive of
(5)
FP tubes/total
no. of gassing
tubes£
61.06
64.50
60.09
E. coli Types
I and II.
£ FP-faloe positives; gassing tubes frost which E. coli was not recovered.
£ Expressed as a percentage.
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believe the uae of 44 C is Justified since the recovery of E. coli and
the enumeration of false positives by the LSI at this temperature
were not significantly different from those by the APHA method. With
Medium A-I, however, a tesperature of 44.5 C was used because this is
the most acceptable teisperature for the incubation of the fecal coli form
group (1). The effects of the direct incubation of LSI and Mediua A-l
over a range of teaperatures will be the subject of a subsequent report.
When the overall performance of Medium A-l was compared with that of
the APHA procedure, results by both methods were aidlor in onuaerating
coliforas (674 and 665) and E. coli (261 and 257). When the tvo rapid
methods were compared to each other, Medium A-l recovered eiore coli-
fonns (674 compared to 631) and E. coli (261 compared to 222) than did
LSI incubated at 44 C. Both rapid tests norit further inventigation
because of the savings in tine and cost of media, without an accompany-
ing loss in accuracy. It would be particularly significant if a rapid,
reliable method could be adopted as a standard cathode procedure for
determining the bacterial quality of shellfish-growing waters as well as
the bacterial content of oysters, claaa, and other potentially narketable
seafoods.
ACKNOWLEDGMENT
We wish to express our appreciation to Jack Gaines and Merrill
McPhearson for collecting saaples and to Bob Huntley and Clark Jones
for cheir very capable technical assistance.
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LITERATURE CITED
1. Aaerican Public Health Association. 1965. Standard methods for
the examination of water and wastewater, pp. 594-600. 12th ed.
American Public Health Association, New York.
2. Edwards, P.R., and W.H. Ewing. 1962. Identification of Enterobac-
teriaceae. Burgess Publishing Co., Minneapolis.
3. Fishbein, M. 1962. The aerogenic response of Escherichia coli
and strains of Aerobacter in EC broth and selected sugar broths
at elevated temperatures. Appl. Microbiol. 10:79-85.
4. Fishbein, M., and B. F. Surkiewicz. 1964. Comparison of the re-
covery of Escherichia coli from frozen foods and nuteasats by con-
firmatory incubation in EC mediua at 44.5 and 45.5 C. Appl.
Microbiol. 12:127-131.
5. Fishbein, M., B.F. Surkiowicz, E.F. Brown, H.M. Oxley, A.P. Pcdron,
and R.J. Groomss. 1967. Col if or ra behavior in frozen foods. I.
Rapid test for the recovery of Escherichia coli frost frozen foods.
6. Raj, H., and J. Lieton. 1961. Detection and enumeration of fecal
indicator organisms in frozen seafoods. I. Escherichia coli.
Appl. Microbiol. 15:233-238.
7. Tennant, A.D., '-.nd J.E. Reid. 1961. Co 1 if or u bacteria in sea water
and shellfish. I. Lactose fernantotion at 35.5° and 44° C.
Canad. J. Microbiol. 7:725-731.
8. Tennant, A.D., J.E. Reid, L.J. Rockwell, and E.T. Bynoe. 1961.
Colifora bacteria in sea water and shellfish. II. The E.C.
confirmation test for Escherichia coli. Canad. J. Microbiol.
7:733-739.
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