A MICROBIOLOGICAL SURVEY IN LAKE  ERIE NEAR CLEVELAND, OHIO
                                by
                         Ralph P. Collins
                  The Biological Sciences Group
                   Regulatory Biology Section
                   University of Connecticut
                   Storrs, Connecticut 06268
                             for the


               Office  of Research and Monitoring

                 ENVIRONMENTAL PROTECTION AGENCY
                       Project #16020  GDQ
                           October 1971
For sale by the Superintendent of Documents, U.S. Government Printing Office, Washington, D.C. 20402 - Price 60 cents

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                  EPA Review Notice
This report has been reviewed by the Environmental Protection
Agency and approved for publication.  Approval does not
signify that the contents necessarily reflect the views and
policies of the Environmental Protection Agency nor does
mention of trade names or commercial products constitute
endorsement or recommendation for use,

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                            ABSTRACT
For several years the Crown Water Treatment Plant in Cleveland, Ohio
has experienced periodic taste and odor problems and the present inves-
tigation was concerned with the role that microorganisms play in this
problem.  During June, July and August of 1971 collections of fungi,
bacteria and algae were made near the intake of the Crown Treatment
Plant.

The studies showed that fungi and bacteria played little, if any, role
in the taste and odor problem at the Crown Plant.  However, a number of
algae which have been reported to induce taste and odor in water sup-
plies were identified in the present study.  Those taste and odor algae
which were found in relative abundance included:  Ceratium sp.,
Coelosphaerium sp., Dinobryon sp., Fragilaria sp., Pediastrum sp.,
Staurastrum sp., Tabillaria sp., and Mougeotia sp.

There was no evidence that benthic organisms played any significant role
in the taste and odor problem experienced at the Crown Treatment Plant.

This report was submitted in fulfillment of Project Number 16020 DGQ
under the sponsorship of the Water Quality Office, Environmental
Protection Agency.
                               iii

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                          CONTENTS
Section




 I           Conclusions




 II          Recommendations




 III         Introduction




 IV          Materials and Methods




 V           Experimental Results




 VI          Discussion




 VII         Acknowledgements




 VIII        References




 IX          Glossary
Page




  1




  3




  5




  7




 11




 25




 27




 29




 31

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                         TABLES

Number   Title                                              Page

   1     Formulation of media used in the isolation           8
         and culture of fungi.

   2     Vertical sampling of plankton flora and fauna       11
         (organisms per liter) taken from Lake Erie at
         the Crown Point Intake, 6-19-71.

   3     Vertical sampling of plankton flora and fauna       13
         (organisms per liter) taken from Lake Erie at
         the Crown Point Intake, 7-17-71.

   4     Vertical sampling of plankton flora and fauna       14
         (organisms per liter) taken from Lake Erie at
         the Crown Point Intake, 8-14-71.

   5     Vertical sampling of chemical and physical data     16
         taken from Lake Erie at the Crown Point Intake,
         6-19-71.

   6     Vertical sampling of chemical and physical data     17
         taken from Lake Erie at the Crown Point Intake,
         7-17-71.

   7     Vertical sampling of chemical and physical data     18
         taken from Lake Erie at the Crown Point Intake,
         8-14-71.

   8     Vertical sampling of fungi taken from Lake Erie     19
         at the Crown Point Intake, 6-19-71.

   9     Vertical sample of fungi taken from Lake Erie       19
         at the Crown Point Intake, 7-17-71.

  10     Vertical sample of fungi taken from Lake Erie       20
         at Crown Point Intake, 8-14-71.

  11     Total coliform count enumerated as organisms        20
         per 100 ml.

  12     Fecal coliform count enumerated as organisms        21
         per 100 ml.

  13     Total plate counts enumerated as organisms          22
         per ml.

  14     Total counts for sulfur, nitrifying and iron        23
         bacteria and total yeast count.
                           vii

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                           SECTION I

                          CONCLUSIONS

1.  Satisfactory sampling procedures for both phytoplankton and benthic
organisms were developed.

2.  It was concluded that bacteria and fungi play little, if any, role
in the taste and odor problem at the Crown Treatment Plant.

3.  No evidence could be found  that benthic organisms play a significant
role in the development of tastes and odors at the Crown Treatment Plant.

4.  Certain phytoplankton collected did produce earthy odors.

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                           SECTION II

                        RECOMMENDATIONS

The problem of taste and odor in Lake Erie and, more particularly, at
the Crown Treatment Plant is undoubtedly complex in origin, and if an
understanding of the problem is to be achieved, a more comprehensive
approach than the one utilized in the summer of 1971 is needed.

This particular problem needs more adequate funding if a thorough job
is to be done.  Collections should be made more frequently and over a
longer period of time than was possible in the present study.

If the problem were continued, a shift in the kinds of personnel
employed would be desirable.  The present study has demonstrated that
bacteria and fungi do not play a significant role, therefore, continued
expertise in these areas is not necessary.  A person skilled in phyto-
plankton work, including skills in identification and culturing, will
still be needed.  It is recommended that an organic chemist broadly
trained in the isolation of trace amounts of organic constituents could
be profitably employed.  A sanitary engineer could be of some help to
the team, particularly if he has knowledge of the area.

It is recommended that at least some of the personnel have sufficient
diving skills to enable them to work in depths of water up to sixty
feet.

It is recommended that a mobile laboratory be placed at the site so
that analyses could be continuous and that time is not lost in trans-
porting samples to adequate laboratories.  The mobile laboratories
should be under the control of the EPA and should be released for
stipulated periods and then returned to a central pool for reassignment.

If it were at all possible, the EPA should also have a facility of this
sort for boats.  One of the amazing things in the recent study was the
fact that boats of the size needed for this work were extremely
difficult to locate.

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                          SECTION III

                          INTRODUCTION

The western suburbs of the city of Cleveland, Ohio have had problems
with taste and odor in the municipal water supply for the past several
years.  This condition was particularly severe during the summer of 1966
and again during the summer of 1967.

The western suburbs of Cleveland are served by the Crown Water Treatment
Plant, which is a rapid sand filter plant with a design filtering capa-
city of 50 mgd.  The raw water intake for the Crown Plant is located
2.5 miles offshore, 46 feet below the surface of Lake Erie.  The circu-
lar crib is 10 feet high and has a diameter of 60 feet.  Initially, the
intake was located four feet above the bottom and at the center of the
crib.

Potos (1) and Kleveno, Braideck and Gehring (2) concluded that most of
the taste and odor complaints occurred when the Crown Plant raw water
intake was located in the hypolimnion.  These same workers showed that
the appearance of a hypolimnion in the vicinity of the Crown Plant was
dependent upon prevailing winds from a southerly direction.  When the
wind direction is southerly, the surface waters of Lake Erie are pushed
to the northern shores with the result that the hypolimnion rises in the
south and becomes depressed in the northern area.

It was assumed that the problem at the Crown Plant could be solved by
moving the intake so that it would always be above the hypolimnion.
However, when the intake was placed above the hypolimnion, it was found
that the taste and odor problems continued.  This suggested that there
was a break in the intake line and investigation revealed that this was
indeed the case.  During the course of this investigation, plans to
repair the break in the intake line were being carried out and it is
assumed that this will solve to a large extent the taste and odor prob-
lem existing at the Crown Plant.  This optimism seems warranted because
other nearby plants which draw their water only from the epilimnion have
not experienced the problems found at the Crown Plant.

The taste and odor problems at the Crown Plant during the summer of 1971
were not particularly severe and this could be attributed to environ-
mental conditions which precluded the formation of a persistent hypo-
limnion.

In the study reported by Potos (1) , sampling was done for the most part
in the vicinity of the Baldwin Water Treatment Plant intake.  The
Baldwin Crib is located 7 miles east of the Crown Plant Crib.  The
Baldwin site has definite advantages because the Crib is raised and
sampling is more reproducible.  In the present study, however, it was
felt that the collections should be made as close to the Crown Crib
as possible and, therefore, all collections were made in the immediate
vicinity of the Crown Crib as well as within the Crib.  Some collections
for bacterial analyses were made in the vicinity of a marker buoy approxi-
mately two miles offshore.

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                           SECTION IV

                     MATERIALS AND METHODS

This section will be divided into three parts:  the first part dealing
with the algae; the second with the fungi; and the third with the
bacteria.

Plankton samples were taken at various depths in the vicinity of the
Crown Point Intake.  Samples initially were taken with a six liter
plankton bottle and immediately concentrated by pouring the samples
through a No. 20 mesh plankton net.  The concentrated material was
collected in a vial attached to the net.  In later experiments the
water samples from each depth were increased to 12 liters.

Benthic algae samples were collected by scuba divers who pushed a
10" x 7" open plexiglass box into the soft muck bottom.  A bottom plexi-
glass plate was slipped underneath the box and the isolated muck
samples brought to the surface.

Periphyton samples were collected from the cement walls and rocks at
the base of the crib.  All samples were fixed in a preservative, stored
in an ice chest, and transported to the laboratory in Storrs, Connecticut,
where final analyses were made.

Water samples, taken at the same depths as the plankton samples, were
collected with six liter plankton bottles for chemical analyses.  The
pH, alkalinity, carbon dioxide, and oxygen concentrations were analyzed
immediately aboard the vessel.  The remaining water from each depth was
stored in glass bottles, placed in an ice chest, and the remaining
analyses carried out in Storrs, Connecticut.

All of the above experiments concerned with chemical analyses were
performed with the Hach Kit.

Plankton counts, in the initial experiments, were determined by a
millipore filter method.  However, this method proved to be unsatisfac-
tory.  All later samples were analyzed by the Sedgwick Rafter Cell
Method.  The plankton were identified and counted using a 12.5x
eyepiece and a lOx objective.  Filamentous algae were counted by cell
counts; colonial and all other species were counted as individuals.
Plankton data are reported as organisms per liter.

The fungi were collected by taking an aliquot of water from the six
liter plankton bottle at each of the depths sampled.  The collected
material was placed in sterile Blake bottles and immediately placed on
ice in an ice chest.  The samples were then analyzed at the Connecticut
laboratories.

Aliquots (2 ml), either diluted or undiluted, were placed on various
media (see Table 1) and then three replicate flasks for each depth were

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incubated at 20C and 25C for varying periods of time.  As soon as
cultures had grown out sufficiently, identifications were made.

The only exception to the above procedure was in those cases where hemp
seed and hair were used to trap aquatic molds.  In those cases one or
two sterile hemp seeds and a few strands of human hair were placed in a
sterile petri dish.  Water collected from one of the sampling depths was
then poured into the petri dishes (approximately 15 ml of lake water
added) and the petri dishes incubated in the usual fashion.  The proce-
dure described above was used for water collected at each depth sampled.

Table 1.  Formulation of media used in the isolation and culture of fungi.
                        Rose Bengal Agar
Ingredients
Neopeptone
Dextrose
Rose Bengal
Aureomycin HCl
Agar
Water
Amounts
5.0 g
10.0 g
0.035 g/1
35.0 g/1
20.0 g
1000.0 ml
                    Sabouraud Dextrose Agar
Ingredients
Neopeptone
Dextrose
Agar
Water
Amounts
  10.0 g
  40.0 g
  10.0 g
1000.0 ml
                      Littman-Oxgall Agar
Ingredients
Neopeptone
Dextrose
Oxgall
Agar
Crystal-violet
Water
Amounts
10.0 g
10.0 g
15.0 g
20.0 g
0.01 g
1000.0 ml
                       Malt Extract Agar
Ingredients
Yeast extract
Malt extract
Neopeptone
Dextrose
Agar
Water
Amounts
   3.0 g
   3.0 g
   5.0 g
  10.0 g
  20.0 g
1000.0 ml

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                      Lindegren Yeast Agar

Ingredients                                                Amounts
Yeast extract                                                 5.0 g
Proteose peptone No. 3                                        3.5 g
Dextrose                                                     40.0 g
Potassium acid phosphate                                      2.0 g
Magnesium sulfate                                             1.0 g
Agar                                                         20.0 g
Water                                                      1000.0 ml
The samples used in the bacterial analyses were collected at the crib
and at a site approximately one-half mile south of the crib.  For the
analysis of aerobic bacteria, a Nisken Sampler was used (3).  For the
analysis of anaerobic bacteria, collections were made by putting approxi-
mately 25 ml of water obtained from the plankton bottle into sterile
one-ounce prescription bottles containing one ml of Bacto-fluid
thioglycollate medium.  These bottles were placed in a desiccator which
was evacuated as soon as the ship reached the shore.  In both of the
situations described above collections were made at the surface and at
depths of 20 and 40 feet.

The Niskin bags, along with the desiccator, were stored in an ice chest
until examined at Storrs.  Sediment samples were collected in the plexi-
glass sampler previously described and the samples for bacteriological
examination were removed with a sterile spatula and placed in sterile
bottles.  These bottles were placed in the desiccator containing the
anaerobic water samples.  Spread plates (4) on Bacto-Plate Count Agar
(5) were incubated at 35C, 20C and 5C for 24 hours, 48 hours and 4
weeks respectively.  Appropriate dilutions were made using sterile dis-
tilled water.  Total and fecal coliform determinations were made on
membrane filters (HAWG, 0.45 u, Millipore) according to "Standard
Methods for the Examination of Water and Waste Water" (6).

The anaerobic samples were plated on plate count agar and the plates
were incubated in an anaerobic jar evacuated with a BBL Gas-Pak at 20C.
Samples were also placed in anaerobic sulfate broth consisting of yeast
extract, 1 g; neopeptone, 1 g; sodium lactate, 8.0 g; sodium sulfite,
0.1 g; ammonium sulfate, 0.1 g; magnesium sulfate, 0.1 g; ascorbic acid,
0.1 g; ferrous ammonium sulfate, 0.1 g; 2 ml of potassium dibasic
phosphate; and 1.0 liter of distilled water.  The media was dispensed
into sterile test tubes, the water sample added and then the tubes were
covered with sterile paraffin and incubated at 20C for 4 weeks.  Water
samples from the Niskin bags were aseptically added to flasks containing
media for nitrifying and iron bacteria.  The media for the nitrifying
bacteria consisted of (NH^^SO^, 200 mg; K2HP04, 50 mg; chelated metals
solution, 1 ml; CaCC>3, 0.3 g; Phenol Red, 1.0 ml of 0.5% aqueous solution;
distilled water, 1000 ml (CoCl2-6H20, 0.004 g; CuSO^Sl^O, 0.0004 g;
FeCl2-6H20; 1.0 g; ZnS04'7H20; 0.3 g; MnS04'H20; 0.6 g; Na2Mo04'2H20,
0.15 g; EDTA, 6 g; made up to one liter with glass-distilled water).
The liquid medium for iron bacteria (Lieoke) consisted of ammonium
sulfate, 1.5 g; potassium chloride, 0.05 g; magnesium sulfate, 0.05 g;
potassium monobasic phosphate, 0.05 g; calcium nitrate, 0.01 g; and

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distilled water, 1000 ml.   These flasks were incubated at 20C for 4
weeks.  At the end of this time period the flasks of Lieoke's medium
were streaked out on Waksman's medium containing ammonium sulfate, 0.5 g;
magnesium sulfate, 0.5 g;  potassium monobasic phosphate, 0.5 g;  sodium
nitrate, 0.5 g; calcium chloride, 0.2 g; agar, 18.0 g and distilled
water, 1000 ml; this media was placed in test tubes and when the plates
were poured 1.0 ml of a 15% solution of ferric ammonium citrate  x^as added.
Samples were also filtered through membrane filters for incubation on
yeast medium consisting of nutrient aear, 2.3 g; glucose, 1.0 g; yeast
extract, 0.1 g; malt extract, 0.2 g; chloromycetin, 100 mg; distilled
water, 100 ml.  These plates were incubated at 20C for one week.
                               10

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                           SECTION V
                      EXPERIMENTAL RESULTS

As in the Materials and Methods section, the results for each group of
organisms will be considered separately and the chemical data will be
included with algae result section.

During the three months of sampling (June-August) no vertical pattern of
distribution was noted in quantitative analysis of the plankton flora.
Wright and Tidd (7) also observed this same phenomenon.  Yearly variation
in the production of phytoplankton have been observed by Chandler and
Weeks (8) in Lake Erie and they attribute this to temperature, rate of
eastward flow of water and nutrients emptying into the lake from streams.

In June the maximum concentration of phytoplankton (organism per liter)
was considerably lower than that observed in July and August.  Maximum
concentration occurred at the surface with 13,674 organisms per liter.
Similar concentrations were observed at the 3 and 9 meter depths.  The
lowest concentrations were observed at the 6 and 12 meter depth (Table 2).

Table 2.  Vertical sampling of plankton flora and fauna (organisms per
liter) taken from Lake Erie at the Crown Point Intake.

                            6-19-71
Organisms per liter
Asterionella
formosa
Colonial greens
Fragilaria
crotonensis
Mougeotia sp.
Pediastrum duplex
Staur_astrum sp.
Tabellaria fenestrata

Total organisms
per liter
Cladocerans
Copepods
Rotifers
Total organisms
per liter
Surface
261
87
9122
3682

522
13,674
87
435
87
3.05M
614
6137
5262
701
87

12,802

87
87
l
609 174
6.10M
174
3819
2752
87

6,832

618

618
9.15M
87
3815
8175
632
270
12,979
87
453
87
609
12.20M
87
6631
998
362

8,078

179

179
13.73M










                               11

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In July the concentration of phytoplankton (organisms per liter) was
considerably higher than that observed in June (Table 3).  The highest
concentration occurred at the surface with 225,682 organisms per liter.
The concentration of phytoplankton decreased progressively with depth
to a minimum of 44,522 organisms per liter at the 12 meter depth.

In August, the total concentration of phytoplankton observed at various
depths was similar to that seen in July.  Maximum concentration occurred
at 12 meters with 208,419 organisms per liter (Table 4).

From June to August, two species of phytoplankton appeared to be dominant
at all depths sampled.  These species were Fragilaria crotonensis and
Mougeotia sp.  The relative abundance of these two species at all depths
sampled ranged from 72% to 96%.  In July and August, Ceratium
hirundinella and Tabillaria fenestrata appeared to increase in numbers
over that observed in June.

The fauna observed during the sampling period of July through August
appeared to be relatively low with Copepods being dominant in June and
Rotifers dominant in July and August.

Based on monthly sampling, the total number of algal species observed
increased from June to August.  The number of species observed increased
from 7 in June to 26 in August.  In June and July, the number of species
appeared to be generally uniform with increased depth.  In August, the
number of species appeared to increase to a depth of 12.20 meters, after
which a decline in species numbers was observed (Tables 4-6).

In terms of the total number of species observed at each depth, the
results of the July and August analysis indicated that the Chlorophyta
(green algae) were the dominant phytoplankton group present, totaling
from 45% to 70% of the community.  During both sampling periods, the
ratio of green algae to total species was less at the surface than that
observed at all other depths.  Similar results were obtained in the June
sampling.

From direct observation and sampling of the bottom mud, no benthic algae
were observed during June through August.  The benthic mud samples were
brought to the laboratory and placed in a culture room in an attempt to
promote growth.  Samples were left in a lighted culture room (100 f.c.)
for a period of a week to ten days during which time no algal growth was
observed.

No periphyton algal community of any significance was observed growing
on the rocks or walls of the water intake crib, although a few clumps
of Cyanophyta (blue-green algae) were observed on the walls during the
August sampling.

Prior to the July sampling of the waters around the Crown Point Intake,
information was received indicating that odor and taste problems were
becoming very obvious in the Cleveland, Ohio area.  In comparing the
phytoplankton data for June and July, no drastic shift in the community


                                12

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Table 3.  Vertical sampling of plankton flora and fauna (organisms per
liter) taken from Lake Erie at the Crown Point Intake,
                            7-17-71
Organisms per liter
Anabaena sp.
Asterionella
formpsa
Ceratiuro
hirundinella
Coelastrum
microporum
Cosmarium sp.
Dictyosphaerium sp.
Dinobryon sp.
Fragilaria
crotonensis
Mi c r o cystics
aeruginpsa
Mougeotia sp.
Cocystics sp.
Pediastrum duplex
Pediastrum simplex
Scenedesmus sp.
Staurastrum sp.
Tabellaria
fenestrata
Unicellular
greens
Total organisms
per liter
Rotifers
Total organisms
per liter
Surface
1089
2397
7848


870
43,829

216
162,241
1308
1743


1525
2616

225,682


3.05M
442
4861


110
1789
20,107

221
120,864
1989
442
884

663
221
1547
154,119
221
221
6.10M
663
3756

110
110
1105
20,770

72,253
1768

221
221
1325
663
663
103,628
221
221
9.15M
221
1989

221
442
1989
10,827

45,959
663
221
110

663
221
110
63,636
110
110
12.20M
442
884

221
994
11,490

28,945
221
442
110

663
110

44,522


13.73M


















                               13

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Table 4.  Vertical sampling of plankton flora and fauna (organisms per
liter) taken from Lake Erie at the Crown Point Intake.
                            8-14-71
Organisms per liter
Anabaena sp.
Asterionella
formosa
Ceratium
hirundinella
Chroococcus sp.
ClqsteriopsJ._s sp.
Coelastrum
microjiprum
Coelosphaerium sp.
Cosmarium sp.
Dictyosphaerium sp.
Dinobryon sp.
Euglena sp.
FragjLlaria
crotonensis
Gleocystics sp.
Kirchneriella sp.
Micractinium sp.
MougeoJ:ia sp.
Oscillatoria sp.
Pediastrum duplex
Pediastrum simplex
Scenedesnus sp.
Sphaerocystics sp.
SjiirofiZia sp.
Stauroneis sp.
Stephanodiscus sp.
Tabellaria
fenestrata

Surface

2279

2127



152



456
304
304
30,382



3342


608



1823

4557


3.05M
166
7623

5303






166
166


77,888
663

331
17,566

497
994


1657
3149

8617


6.10M
152
1215

1823


304


456
152

2734
152
65,321



12,760

304
1671
152


2582

6076


9.15M
304
8507

4405


304
152

152

304
152

136,718

152

21,875
456
456
706

2734

5317

6988


12.20M
152
5772

9418

304
456
152

152
152
1215


126,388

152

36,762
152
152
2734
152
608

8811

14,279


13.73M

1975

3646


152




152


67,751



16,406

152
304

304

2582
152
1215


                               14

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Unicellular greens
Total organisms
per liter
Rotifers
Total organisms
per liter

46,486



124,786
166
166
152
96,006



190,192
1062
1062

208,419
304
304

94,791


composition was observed which may have caused the problem, although
Ceratium hirundinella was found at the surface in large numbers (7848
organisms per liter).  It has been well documented that this species is
associated with coloring of waters of reservoirs and lakes, and causes
taste and odor problems in the same, although the very significant
increase in the previously existing phytoplankton may have also been a
major factor.

Several of the species isolated in this study, e.g., Ceratium sp.,
Coelosphaerium sp., Dinobryon sp., Fragillaria sp., Pediastrum sp.,
Staurastrum sp., and Tabillaria sp., have been reported as important
taste and odor producing algae by Palmer (9, 10, 11, 12).  While Mougeotia
has not been reported by Palmer as an important taste and odor producing
alga, its presence in large numbers in the present study suggests that
it may also be involved in the production of undesirable tastes and odors.

While realizing the limitations of information gathered by means of the
Hach Kit, nevertheless, the data does tend to substantiate the findings
made by other workers working in the same general region.

The fungi isolated on the various media are listed in Tables 8, 9 and 10.
None of the fungi isolated produced an earthy odor of any sort even
though they were incubated, in most cases, for several weeks.  One
actinomycete culture was isolated in the August sampling and this
organism did produce an earthy-musty odor.

The predominant fungi collected were various yeasts and species of
Penicillium, Aspergillus and Alternaria.  None of these cultures produced
an odor.  One isolate of Streptomyces was found and it produced an earthy-
musty odor.  However, as this organism was isolated but once, it is hard
to visualize the actinomycetes playing any significant role in the odor
problem at the Crown Plant.

An examination of the bottom mud aid scrapings from the wall of the Crib
revealed no taste and odor producing microorganisms although an inter-
esting species of Phoma was isolated from material collected in the Crib.
                              15

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Table 5.  Vertical sampling of chemical and physical data taken from
Lake Erie at the Crown Point Intake.
                            6-19-71
PPM
Alkalinity (CaC03)
Bicarbonate
Carbonate
Total
Hardness (CaCO.,)
Calcium
Magnesium
Total
Iron
Nitrate Nit.
Nitrite Nit.
Oxygen
Ph
Phosphate
Ortho
Total
Sulfate
Temperature (C)
Turbidity
(JT Units)
Surface

80.0
10.0
90.0
100.0
21.0
121.0
0.05
0.13
0.01
11.0
8.60

0.04

21.0
21.0
0.0
3.05M

90.0
10.0
100.0
90.0
37.0
127.0
0.05
0.15
0.01
11.5
8.60



22.0
20.0
0.0
6.10M

86.0
14.0
100.0
95.0
30.0
125.0
0.0
0.11
0.008
12.0
8.60

0.03
0.11
21.0
19.5
3.0
9.1511

100.0
0.0
100.0
89.0
30.0
119.0
0.02
0.09
0.004
7.0
7.70

0.01
0.14
18.0
15.0
8.0
12.20M

99.0
0.0
99.0
90.0
30.0
120.0
0.02
0.09
0.005
6.5
7.60


0.14
18.0
11.8
2.0
13.73M


















                              16

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Table 6.  Vertical sampling of chemical and physical data taken from
Lake Erie at the Crown Point Intake.

                            7-17-71
PPM
Alkalinity (CaC03)
Bicarbonate
Carbonate
Total
Carbon Dioxide
Hardness (CaCO-j)
Calcium
Magnesium
Total
Iron
Nitrate Nit.
Nitrite Nit.
Oxygen
Ph
Phosphate
Ortho
Total
Sulfate
Temperature (C)
Turbidity
(JT Units)
Surface
65.0
20.0
85.0
0.0
93.0
32.0
125.0
0.0
0.109
0.010
8.70
8.58

0.004
0.005
20.0
21.0
0.0
3 . 05M
65.0
20.0
85.0
0.0
94.0
31.0
125.0
0.0
0.114
0.009
8.50
8.58

0.003
0.005
18.0
21.0
0.0
-
6.10M
62.0
20.0
82.0
0.0
95.0
31.0
126.0
0.0
0.115
0.100
8.60
8.50

0.003
0.004
20.0
21.0
0.0
9.15M
64.0
20.0
84.0
0.0
94.0
36.0
130.0
0.0
0.113
0.100
8.90
8.54

0.003
0.005
20.0
21.0
0.0
12.20M
68.0
20.0
88.0
0.0
92.0
29.0
121.0
0.0
0.113
0.100
8.90
8.43

0.003
0.004
19.5
20.9
0.0
13.73M
67.0
20.0
87.0
0.0
98.0
22.0
120.0
0.0
0.118
0.009
8.60
8.51

0.004
0.005
19.5
21.5
0.0
                               17

-------
Table 7.  Vertical sampling of chemical and physical data taken from
Lake Erie at the Crown Point Intake.

                            8-14-71
PPM
Alkalinity (CaCO^)
Bicarbonate
Carbonate
Total
Carbon Dioxide
Hardness (CaCO-j)
Calcium
Magnesium
Total
Iron
Nitrate Nit.
Nitrite Nit.
Oxygen
Ph
Phosphate
Ortho
Total
Sulfate
Temperature (C)
Turbidity
(JT Units)
Surface

87.0
10.0
97.0
0-1

100.0
25.0
125.0
0.0
0.024
0.006
11.0
8.54

0.005
0.130
20.0
22.0
1.0
3.05M

80.0
20.0
100.0
0-1

97.0
33.0
130.0
0.0
0.023
0.006
12.5
8.25

0.003
0.003
23.0
22.0
1.0
6.10M

75.0
20.0
95.0
0-1

99.0
22.0
121.0
0.0
0.025
0.005
9.0
8.50

0.006
0.006
24.0
22.7
8.0
9.15M

93.0
10/0
103.0
0-1

100.0
30.0
130.0
0.0
0.026
0.004
10.5
8.45

0.003
0.170
23.0
21.2
1.0
12.20M

80.0
10.0
90.0
0-1

100.0
20.0
120.0
0.0
0.023
0.007
13.0
8.20

0.005
0.100
23.0
22.6
0.0
13.73M

90.0
0.0
90.0
0-1

100.0
30.0
130.0
0.0
0.05
0.009
8.0
7.30

0.006
0.120
20.5
21.2
10.0
                               18

-------
Table 8.  Vertical sampling of fungi taken from Lake Erie at the
Crown Point Intake.

                            6-19-71
Average No. of
Colonies per
Petri dish
Alternaria tenuis
Penicillium sp.
Aspergillus niger
Monila sitophila
Aureobasidium
pullulans
Sporojtjrichura sp.
Saccharomyces
cerevisial
Rhodpjxmila
Surface
13
5
8
4
10

8
2

3
3.05M
5
3
5
1
5

2
1

2
6.10M
2
1
3



1


1
9.15M



1
3

1


1
12.20M


1






1
13.73M










Table 9.  Vertical sanple of fungi taken from Lake Erie at the Crown
Point Intake.
                            7-17-71
Average No. of
Colonies per
Petri dish
Aureobasidium
pullulans
R^qt^ojruLa sp.
Alternaria tenuis
Penicillium sp.
Aspergillus niger
Monila sitophila
Candida sp.
Geotrichum
Candida
Spprotrichum sp.
Cladpsjiorium sp.
Tusarium sp.

Surface
10

8
3
2
5
2
4
2

1
1
4
3.05H
6

7
2
1
8
1
1
1

1
2
1
6.101!
4

6
1
1
2

1



1

9.15M
1

2

1
1

1
2

1

1
12.20M


1
1
2



1


1

13.73!!


1

1


1





                               19

-------
Table 10.  Vertical sample of fungi taken from Lake Erie at Crown Point
Intake.
                            8-14-71
Average No. of
Colonies per
Petri dish
Aureobasidium
pullulans
Rhodotorula sp.
Aspjargillus sp.
Penicillium sp.
Candida sp.
Geotrichum sp.
Alternasia tenuis
Tusarium sp.
Paecilomyces
elegans
Gliocladium sp.
Streptomyces
Surface
8

10
2
4
3
2
5
1
1

2
1
3.05M
4

8
1
2
1
1
5
2
1



6.10M
4

7
1
2
2
2
4
1


1

9.15M
3

5
2

2
1
3
1




12.20M


4
1


1




1

13.73M
1

1


1







The information gathered from the analyses for bacteria are discussed below.

The total coliform counts are shown in Table 11.  The results designated
as A in the table were made at the Crown Point Intake while those desig-
nated as B in the table were taken at a sampling site one-half mile from
the Crib.  The designation S, as in A-S, refers to the surface sample
while the number, for example A-20, refers to depth at which the collec-
tion was made.  A-sd refers to the sediment sample.

Table 11.  Total coliform count enumerated as organism per 100 ml.

Station
A-S
A-20
A-40
A-sd
B-S
B-20
C
June
3650
3600
1950
NT
16,000
>io
;ollection Period
L JulY
255
240
253
700
4100
310

August
10
30
30
100
30
140
                               20

-------
In Table 12 the fecal coliforms are shown.  The designation NT means
not taken.
Table 12.  Fecal coliform count enumerated as organisms per 100 ml.
S_tation_

A-S
A-20
A-40

A-sd

B-S
B-20
B-40
Collection Period
June
112
38
75
NT
15
10
NT
July
>2
>2
>100
10
>2
NT
August
>]
1
^100
3
8
16
The total plate counts at the various incubation temperatures are shown
in Table 13.  The designation NT means not taken.
In Table 14 the counts for sulfur, nitrogen and iron bacteria are
expressed as well as the total yeast counts.  In the portion of the
table concerned with the sulfur bacteria the numbers indicate how much
sample was added to the medium in order to get a positive test reaction.
A positive reaction being blackening in the sulfate broth medium.  If,
for example, we look at station A-S for the three sampling periods, we
find that a positive reaction for sulfur bacteria was noted in June and
July when 10 ml of the sample was added to the medium.  In August the
reaction was negative for all of the samples analyzed.  In the A-sd
sample we find that in July there was a positive reaction when 1 ml of
a 1/100 dilution of the sample was used and in August we find a positive
response when 0.01 ml of a 1/100 sample was tested.

In the section of the table concerned with nitrifying bacteria the same
scheme prevails; for example, in B-S we find the number +200 which means
that 200 ml of the sample material was necessary in order to achieve a
positive response which in this case was a change in pH.

In the section of the table dealing with iron bacteria all of the pour
plates containing Waksman's medium were negative.  Duplicate pour plates
containing either 1.0 or 0.1 ml of sample were used.  In July and August
Lieoke's medium was used to enhance growth and then the organism which
grew out were streaked on Waksman's medium.  The asterisk denotes a posi-
tive response.  A positive test being growth on Waksman's medium.  In the
final section of Table 14 the yeast counts per 100 ml of medium are given
for July and August.  This procedure was not performed in the June sample.
                               21

-------
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                                                                  33
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                                                                   3
22

-------
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                                   25
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                                                                                             CL
ota
yeast coun
                        23

-------
                           SECTION VI

                           DISCUSSION

The present investigation was concerned primarily with a microbiological
survey of the Crown Point Station.  The three major groups of micro-
organisms studied were the fungi, bacteria and algae.  The results of
this study show that the fungi and bacteria play little if any role in
the taste and odor problem present at the Crown Station Inlet.  While
a fair diversity of both fungi and bacteria were noted, the total
counts were not abnormally high nor did individual cultures exhibit any
pronounced odors.

Significant progress was made in identifying and enumerating the species
of the algal community associated with the Crown Point Intake.  Many of
the species observed have been associated with taste and odor problems
in other aquatic environments.  During the period of this study no pro-
nounced taste and odor difficulties were experienced at the Crown Plant.
However, various local people remarked during the August sampling period
that the water had what they described as a typical "Lake Erie odor".
Whatever the source of these odors, they were not due to benthic or peri-
phyton algae, but they could have been associated with the phytoplankton
community within the area as the reported "Lake Erie odor" coincided with
the increase in phytoplankton.

From the results of this survey, it is apparent that a continuation of
the study should be continued.  Much more intensive work is needed in
the field to more rigorously characterize the physical, chemical and
biological relationships of the area, through which patterns can be
developed and models for prediction of cause and effect relationships
of taste and odor problems in Lake Erie.  These prediction models will
enable the filtration plants to prepare for the problems and counter
them when they arise.

Intensive laboratory work is needed to culture species of the phyto-
plankton community found around the Crown Point Intake.  This culturing
should be done with the natural chemical and physical environment in
mind.  Thus, stimuli of phytoplankton growth can be detected, especially
taste and odor species, and controls may be found.  Culturing should
also be used to identify chemical by-products of algae which cause the
taste and odor problems and possible chemical controls  may be applied.
                               25

-------
                          SECTION VII

                        ACKNOWLEDGEMENTS

The collaboration of Dr. Eugene Hansmann (Algologist), Hiss Marjorie
Berry (Bacteriologist), and Mr. David Rathke (Algologist and Diver) are
acknowledged with sincere thanks.

The support of Dr. Ronald Webb, the Grant Project Officer, and the person
who called our attention to this problem, is gratefully acknowledged.
                                27

-------
                           SECTION VIII

                            REFERENCES

 1.   Potos, C.,  "A Study of Taste and Odor in the Municipal Water Supply
     at Cleveland, Ohio," Proceedings of the 31th Conference, Great
     Lakes Research, pp  571-584 (1968).

 2.   Kleveno, C.  0., Braideck, T. E., and Gehring, P. E., "Hypothesis for
     Dissolved Oxygen Depletion in the Central Basin Hypolimnion of Lake
     Erie," U.  S. Department of the Interior, Federal Water Pollution
     J^J15JLO 1 Administration, Great Lakes Region,_ Lakg_ Erie Basin Office,
     pp  1-6 (1970).

 3.   Niskin, S.  J., "A Water Sampler  for Microbiological Studies,"
     Deep-Sea Research, 9^ pp 501-503 (1962).

 4.   Buck, J. D., and Cleverdon, R. C., "The Spread Plate as a Method for
     the Enumeration of Marine Bacteria," Limnology Oceanography, 5,
     PP 78-80 (1960).

 5.   Geldreich,  E. E., "Fecal-Coliform-Organism Medium for the Membrane
     Filter Technique," Journal of the American Water Works Association,
     57, pp 208-214 (1965).

 6.   APHA, AWWA,  and WCCF, "Standard  Methods for the Examination of Water
     and Wastewater," (12th ed) American Public Health Association, Inc.,
     New York,  New York (1965).

 7.   Wright, S.,  and Tidd, W. N. , "Summary of Limnolopical Investigations
     in Western Lake Erie in 1929 and 1930," Transactions of American
     Fisheries Society, 63, pp 271-285 (1933).

 8.   Chandler, D. C., and Weeks, 0. B., "Limnological Studies of Western
     Lake Erie.   V.  Relation of Limnolopical and Meteorological
     Conditions to the Production of  Phytoplankton in 1942," Ecological
     Monographs,  15, pp 435-456  (1945).

 9.   Palmer, C.  M., "Algae and Other  Interference Organisms in Water
     Supplies of California," Journal o_f American Water Works Association,
     5_3_,_ No. 10,  pp 1297-1312 (1961).

10.   Palmer, C.  M., "Algae and Water  Supplies in the Sao Paulo Area,"
     The Robert A. Taft Sanitary Engineering Center, Technical Report
     W61-30, Cincinnati, Ohio (1961a).

11.   Palmer, C.  M., "Algae in Water Sunplies of Ohio," The Ohio Journal
     of Science,  62, No. 5, pp 225-244 (1962).

12.   Palmer, C.  M., "Nuisance Algae in Water Supplies of the Pulp and
     Paper Industry,1' Tappi, 45, No.  12, pp 897-900 (1962a).
                                 29

-------
                           SECTION IX

                            GLOSSARY

Actinomycetes - Filamentous bacteria.

Algae - Chlorophyll-containing plants lacking roots, stems or leaves.

Fungi - Chlorophyll-lacking plants which have no roots, stems or leaves.

Chrysophyta - A group of algae characterized by the formation of a yellow-
brown pigment.

Chlorophyta - A group of algae characterized by the formation of green
coloring materials primarily chlorophyll.

Cyanophyta - A group of algae characterized by blue-green coloring
materials.
                               31

-------
     4ccesj>ioti Number
                             Subject Field &. Group
                               016C
                                                SELECTED WATER RESOURCES ABSTRACTS
                                                       INPUT  TRANSACTION  FORM
     Organization
      The Biological Sciences Group, Regulatory Biology Section
      University of Connecticut,  Storrs, Connecticut   06268
     Title
      A MICROBIOLOGICAL SURVEY IN LAKE ERIE NEAR CLEVELAND, OHIO
 10
     Authors)
      Collins, Ralph P.
                                16
Project Designation

EPA  WQO  Project No. 16020 GDQ
                                     21
                                         Note
 22
     Citation
     Descriptors (Starred First)
 25
Identifiers (Starred First)

 Aquatic microbiology
 27  Abstract
~^ The taste and odor constituents produced by the actinomycete Streptomyces  odorifer,
     the alga Synura petersenii  and the mold Trichoderma  viride were examined.

     The odorous constituents were obtained by steam distillation of the culture
     medium.   The odorous constituend were identified by  means of gas-chromatography,
     infrared, mass and nuclear  magnetic spectroscopy.  The major odorous constituend
     produced by the above named organisms have been enumerated.  (Collins-U.Conn.)
Abstractor
        Ralph P. Collins
                               Institution
                                    University of Connecticut
 WR 102 (REV JULY 1969)
 WRSIC
                                           SEND TO- WATER RESOURCES SCIENTIFIC INFORMATION CENTER
                                                  U S DEPARTMENT OF THE INTERIOR
                                                  WASHINGTON. D  C  20240
                                                                                 CPO: 1969-359-339

-------