DUDLEY KNOX LIBRARY
NAVAL POSTGRADUATE SCHOOL
d POSTGRADUATI MONTEREY, CA 93943.5101

SY. CALIF. 93£

This document has been approved for public
release and sale; its distribution is unliuatea.

h /±

A COLIFORM BACTERIA SURVEY OF
MONTEREY BAY OFF DEL MONTE BEACH

by

David Sidney Trumbauer
Lieutenant, United States Navy
B. S„, University of Maryland, 1960

Submitted in partial fulfillment
for the degree of

MASTER OF SCIENCE IN PHYSICAL OCEANOGRAPHY

from the

UNITED STATES NAVAL POSTGRADUATE SCHOOL

October 1966

\a\o(p

Determination of the distribution of coliform
bacteria in the vicinity of the outfall effluent of
the Monterey Water Pollution Control Plant was made
using the membrane filter technique for enumeration.
The coliforms were seen to follow the onshore mass
transport of water in the surface layer in response
to predominately westerly winds. The extent of
penetration of coliforms into the bay north of the
outfall is restricted to several hundred yards,
while more extensive spreading was observed towards
the surf zone. No consistent pattern of variation
of the coliform distribution could be related to the
tidal cycle. Higher concentrations of coliform bacteria
on several occasions seemed to be related to reduced
periods of solar radiation.

DUDLEY KNOX LIBRARY

NAVAL POSTGRADUATE SCHOOL

MONTEREY, CA 93943-51 01

TABLE OF CONTENTS

Section Page

1. Introduction 7

2. Means of Enumeration of Coliform Bacteria 15

3. Collection of Coliform Data 19

4. Results 23

5. Conclusions 46

6. Acknowledgements 48

7. Bibliography 49

v- X0MMY3JC

LIST OF TABLES

Table Page

1. Summary of weather and tide data for

each coliform survey 24

2. Sampling and inoculation times 25

3. Coliform concentrations observed

along Del Monte Beach 27

4. Coliform concentrations determined on

survey 14 on 18 August 1966 31

5. Coliform concentrations determined on

survey 15 on 23 August 1966 34

6. Coliform concentrations determined on

survey 16 on 25 August 1966 35

7. Coliform concentrations determined on

survey 17 on 27 August 1966 36

8. Coliform concentrations determined on

survey 18 on 30 August 1966 38

9. Coliform concentrations determined on
survey 19 on 1 September 1966

40

10. Coliform concentrations determined on

survey 20 on 3 September 1966 43

LIST OF ILLUSTRATIONS

Figure Page

1. Chart of region surveyed 11

2. Location of stations used in coliform
bacteria survey 13

3. Distribution of coliform bacteria on

18 August 1966 (survey 14) 32

4. Composite distribution of coliform bacteria
for period 23-27 August 1966 (surveys 15 ,

16, 17) 37

5. Distribution of coliform bacteria on

30 August 1966 (survey 18) 39

6. Distribution of coliform bacteria on

1 September 1966 (survey 19) 41

7. Distribution of coliform bacteria on

3 September 1966 (survey 20) 44

1. Introduction.

The purpose of this study was to determine the concen-
tration and distribution of coliform bacteria in the southern-
most portion of Monterey Bay and to investigate qualitatively
the influence of various meteorological and oceanographic
parameters on this distribution.

Coliform bacteria are defined as aerobic and facul-
tatively anaerobic non-spore-forming bacilli that ferment
lactose to produce gas in 48 hours at a temperature of
35° -37° Centigrade. Coliforms refer to a group of bacteria
which are normal inhabitants of the colon or large intestine
of man and most vertebrate animals; as such, they serve as
an indication of fecal pollution in estimating the sanitary
quality of the various waters. This group of bacteria is
composed of two genera, which have nearly identical micro-
scopic and macroscopic morphological properties, and are
distinguishable only by the type of fermentative action
used in cleavage of glucose. Each of these genera are in
turn broken down further into species primarily on the
basis of biochemical reactions. The recognized species of
coliforms are as follows [lj:
Escherichia coli
Escherichia aurescens

Escherichia freundii

Escherichia intermedia

Aerobacter aeroqenes

Aerobacter cloacae
Although all of these are found frequently in the intestine
of vertebrates, only E. coli is strictly an intestinal
organism and the other species have been found associated
with soils and grains as well as in intestinal habitats.
Other organisms have been suggested as alternative indices
of pollution but have been found less satisfactory for wide-
spread use than the coliform index [2] .

The source of coliforms in the area surveyed is the
effluent of the City of Monterey's Water Pollution Control
Plant which is a primary incomplete sewage treatment plant.
The outfall pipe extends northward into Monterey Bay for
800 feet from the shoreline, turns towards the surface, and
terminates approximately 18 feet below the surface. The
point where the effluent reaches the surface may be deter-
mined visually by the turbulence and discoloration at the
surface. The volume being discharged varies from a maximum
between the hours of 0900 and 1500 to a minimum between
0100 and 0600.

8

No other sources of coliforms were considered in this
study, although increases in coliform concentration were
observed at positions remote from the outfall boil.

Once discharged in the sea the concentration of
coliforms decreases rapidly. Tibby, in an investigation
in Santa Monica Bay, established a value for Tqq (the
time required for 90% disappearance of coliforms) of
four to seven hours j[3jl . The reasons for this rapid decline
in concentration are:

(1) The sea is a toxic environment for coliforms. In
the surface layer bacteria are quite sensitive to ultra
violet radiation. Bacteriophages (bacterial viruses) and
antibiotic substances found dissolved in sea water are also
toxic to bacteria. In addition, there is a toxicity of sea
water itself which seems to be associated with the dissolved
solids. This has been demonstrated with artificial sea
water of several formulations£4J . The mechanism of this
toxicity for coliforms is largely unknown.

(2) Coliforms are subject to predation by planktonic
organisms.

(3) The bacterial cells are subject to sedimentation
with particulate matter.

(4) Concentration of coliforms decreases as the
effluent is diluted with sea water.

From the above considerations it is obvious that
the concentration of coliform bacteria are a non-conserva-
tive property in the sea and as such are not meaningful
in quantitative relationships in the sea. However, they
may be used to illustrate qualitative features of the
circulation.

The region being surveyed is located in the extreme
southern portion of Monterey Bay extending along the
beach from Monterey Municipal Wharf Number 2 to the oil
tanks to the east and extending out into the bay approxi-
mately 500 yards from the shoreline (figure 1). The
beach is sandy and of gentle profile. Bottom contours
closely parallel the shoreline. The configuration of the
coastline and the bottom contours is such that the area
surveyed is largely protected from wind wives and swell
approaches the beach with wave crests nearly parallel to
the shoreline. The sector along the beach is also one of
diverging wave orthogonals which tends to disperse wave
energy for most swell directions.

10

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30

26

13

25

vS *0Q ' '"/•••i/,?i„nM|M>'\',,M V60'

y

Yarcis

1000

l-l rt m -I ri

Figure 1 . Chart of region surveyed.
(Taken from C.&G-.S. Chart Sk-03)

Several investigations have been made into the circula-
tion of water in this region. Brennan and Me aux fs] observed
a dominant onshore flow outside the surf zone with onshore
winds and a flow to the north along the beach with frequent
rip currents. Stevenson [6] in investigating the mass
transports around the outfall effluent found that water
of all levels down to 15 feet closely follows the wind. No
consistent circulation pattern could be related to any
section of the tidal curve.

The locations of the stations used in the individual
surveys are shown in figure 2. The stations were placed at
200 yard intervals along the beach and at 100 yard intervals
extending from the beach. Stations 2, 14, 26, 38, 60 and 62
are in line with the outfall pipe. Station 38 is at the boil.
Kelp beds are fairly dense to the west of the outfall boil,
sparse to the north of the outfall boil, and absent to the
east and shoreward of the boil.

Observations made during this survey include coliform
concentration, wind speed and direction, cloud cover, tide
level and state, and sea surface temperature. The means of
coliform enumeration is discussed in the next section. Wind
speeds and directions were measured with a portable anemometer

12

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near station 5 at a height of ten meters above sea level.
Sea surface temperatures were measured at several stations
on each survey and averaged. Cloud cover was estimated
visually. Tidal information was obtained by examination
of the tide gauge records maintained by the Meteorology
and Oceanography Department of the U. S. Naval Postgraduate
School.

14

2. Means of Enumeration of Coliform Bacteria.

Two procedures for the enumeration of coliform bacteria
were considered; the fermentation tube or most probable
number (MPN) method, and the membrane filter (MF) method.

The MPN method was adopted as the official method of
determining coliform densities in the 1936 edition of
Standard Methods. It consists of planting several known
volumes of a water sample into three or more replicates of

1

fermentation tubes partially filled with either lactose or
lauryl tryptose broth. The tubes are observed for the
production of gas after 24 and/or 48 hours. By tabulating
all results for each sample, the MPN or the density of
bacteria most likely responsible for that combination of
results can be determined by referring to MPN tables as
presented in Standard Methods \f] . There are several
practical disadvantages to this method.

(1) A large amount of time is required for glass-
ware and media preparation. A minimum series for three
dilutions and three replicates requires nine tubes of
media in addition to dilution blanks for each sample.

(2) Planting of media, reading, and recording
results is a slow process with numerous opportunities
for labeling-type errors.

15

(3) Presumptive results take 24 to 48 hours and
completed results take at least 96 hours after planting.

(4) Large laboratory facilities are required for
a large number of samples.

(5) Any bacteriocidal agents in the sample are
carried over into the fermentation tube.

The membrane filter procedure for the examination
of water for coliform bacteria was accepted by the
American Water Works Association as an alternate to
the MPN procedure in November 1957 £eQ, and it is described
as a standard test for the coliform group in the 12th
edition of Standard Methods jY] . In the membrane filter
method, a known volume of a water sample is drawn through
a membrane filter prepared from cellulose esters with
pores just smaller than the bacterial cells being sought.
After filtration of the sample the filter is placed on an
absorbant pad saturated with a nutrient preparation which
is selective for the organisms being sought, in this case
coliforms. Each living cell trapped on the filter multiplies
upon incubation at an appropriate temperature and produces
a macroscopic colony of bacteria which may be counted. All
such colonies are counted and the coliform concentration is

16

simply the number of colonies divided by the volume
filtered.

The advantages of this system may be listed as
follows:

(1) A minimum of glassware is required. The filters
can be incubated in disposable petri dishes.

(2 ) By use of a vacuum pump a 50 ml sample can be
filtered in about 30 seconds.

(3) Initial results are available in 24 hours and
completed results may be obtained in an additional 48 hours.

(4) Little space is required for a large number of
samples. The petri dishes are 47 mm in diameter and may
be stacked in columns to conserve space.

(5) Toxic environment in the sample is removed by
filtration.

There are also disadvantages to the MF method.

(1) A fairly good estimate of the coliform concentra-
tion is needed beforehand in choosing an increment of volume
to be filtered. Too many colonies on a single filter give
indeterminate results.

(2) The MF method gives densities which are generally
lower than those determined by the MPN method.

17

Jannasch and Jones [9J , in evaluation of several methods
of enumeration including the MPN and MF methods, observed
MPN densities on the average to be an order of magnitude or
more higher than MF densities. They (Jannasch and Jones £9] ) :.
attribute the larger concentrations obtained with the MPN
method to dispersion of bacterial aggregates and clumps on
foreign matter produced by surface tension depression of the
liquid media. Also, theoretical considerations and large
scale replicate determinations indicate that MPN tends to be
greater than the actual number £7J . Thus, it appears that the
MF method tends to produce results which are biased on the low
side while the MPN method is biased on the high side, and
the results are not directly interchangeable.

18

3. Collection of Coliform Data.

Observations were made on 20 surveys taken during the
period 22 June 1966 through 3 September 1966. The first 13
of the surveys were confined to the surf zone along station
1 westward. The last seven surveys were made in the offshore
region. Samples were collected in sterile 150-200 milliliter
glass bottles with metal screw caps. All samples were hand
shaken 30 times before increments were withdrawn for planting.
All completed cultures were submitted to the differential
tests known as the IMViC tests for the production of indole
from tryptophane broth, the methyl red test for production of
acid, the Voges-Proskauer test for the production acetyl
methyl carbonol, and the use of citrate salts as a sole carbon
source. Standard Methods (V) , was used as a guide in the
preparation of the IMViC reagents, media, and in the execution
of the tests. All media, glassware and equipment except the
membrane filters, Endo media, and disposable petri dishes were
sterilized by autoclaving for 15 minutes at 15 pounds pressure,
The excepted items were sterilized in accordance with instruc-
tions from the manufacturer.

19

With the MPN procedures three replicates of four
dilutions (1 ml, 0.1 ml, 0.01 ml, and 0.001 ml) of the sea
water samples were planted into Difco dehydrated lactose
broth with fermentation tubes and incubated at 37°C for
24 and 48 hours. For increments of sample less than 1 ml,
dilutions were made with sterile phosphate buffered distilled
water. The appearance of gas after 24 or 48 hours was
recorded as positive and the absence of gas after 48 hours
was recorded as negative. Representatives of the highest
dilution giving positive results were confirmed on eosin
methylene blue (EMB) agar. Colonies exhibiting the typical
coliform morphology were inoculated into lactose broth with
fermentation tubes. Nutrient agar slant cultures were made
from positive lactose broth tubes and Gram stains were made.
This procedure constituted the completed test for the presence
of coliform bacteria£7j . The MPN procedure was suspended
after 14 July in favor of the MF procedure.

In the MF procedure the Millipore bacteriological
analysis kit2 was used. A vacuum-pressure pump is used

1 Difco Laboratories Incorporated, Detroit, Michigan

2 Millipore Filter Corporation, Bedford, Massachusetts

20

to facilitate the filtration. Millipore MF type HA filters
with pore size of 0.45* 0.02 microns were used. Various volumes
ranging from 100 to 0.01 ml were filtered. The 100 ml volumes
were found unsatisfactory because of overgrowth of non-coli-
form types. Volumes of 50 ml were used for most stations remote
from the outfall boil. Volumes of 50 and 1 ml were used in the
vicinity of the boil and 0.1 and 0.01 ml were found satis-
factory for the samples from the outfall boil (station 38)
and the station shoreward from the boil (station 26). When
volumes of less than 25 ml were used, 30 to 50 ml of sterile
buffered dilution water was poured into the filtering
funnel prior to adding the sample to provide an even distri-
bution. After filtration, the sides of the funnel were rinsed
with 30-50 ml of sterile buffered dilution water, the filter
removed and placed grid side up on an absorbant pad saturated
with 1.8 to 2.0 ml of freshly prepared M Endo Broth MF"* in
a petri dish, covered, and incubated at 37°C for 24 hours.
Between samples, the funnel was immersed in boiling water for
10-15 seconds. (As evidenced by control filtrations of
sterile water which were run periodically, it was difficult
to remove bacteria from the sides of the funnel even with
repeated rinsings. The immersion in boiling water between

3 Baltimore Biological Laboratories, Baltimore, Maryland

21

samples eliminated this cross contamination.) Coliform
bacteria on Endo media form circular, convex colonies
with a greenish metallic sheen in reflected light. The
growth of non-coliform organisms is inhibited; those that
do grow lack the characteristic sheen. After incubation,
the sheen colonies were counted with the aid of ten power
magnification. Non-coliform colonies were counted separ-
ately to indicate in the results the degree of overcrowding
on the filters. Representative sheen colonies were selected
and inoculated into lactose broth fermentation tubes.
Nutrient agar slant cultures were made from the positive
lactose tubes and Gram stains were made from the slant
cultures. This procedure constituted the completed test
for the membrane filter cultures.

22

4. Results.

The weather and tide observations for each survey
are presented in Table 1. The actual times for sampling
and inoculation are listed in Table 2. The coliform
concentrations, per 100 ml, are listed in Table 3 for
surveys 1 through 13 which were conducted along the
beach. Tables 4 through 10 give the coliform concentra-
tions determined from samples collected seaward of the
surf zone.

Discussion of Surveys 1 through 13

The general pattern of the coliform distribution
along the beach appears to be a region of maximum coliform
concentration at or to the east of station 1 with the level
remaining high to the outfall and a rapid decrease to the
west of the outfall. The occasional higher coliform
densities that were observed remote from the outfall may
possibly be due to sources other than the sewage effluent.
Surveys 5, 6, 9, 11, 12 and 13 all showed increasing coli-
form densities toward Monterey Municipal Wharf No. 2.

Surveys 1, 2, 4, 6 and 12 exhibited local maximums
between stations 4 and 7. Since offshore samples were not
taken on these surveys it is not known if these maximums

23

TABLE 1

SUMMARY OF WEATHER AND TIDE DATA
FOR EACH COLIFORM SURVEY

Survey

Date

Time*
^PDT)

Wind
DIR/KTS

Clouds
(tenths !

Temp,
I (°C)

Tide level
(feet/state)

1

22 JuN

1>30

290°/10

0

16.1

Li.. 1 /LHW

2

25 JtTN

llj.00

290°/1 2

0

16.5

2.1 /Rising

3

6 JUL

1500

275°/12

V

I5.il-

3.6/Falling

k

12 JUL

1230

355°/i 0

1

1V.6

2. 2/HLW

5

1ij. JUL

1I4.00

285°/1ii.

0

18.v

2.7/HLW

6

11|. JUL

2000

285°/1l}.

0

16.8

5. v/hhw

7

, . 2 AUG

1I4.00

2v5°/i5

10

16.1*.

ij.. 1 /Falling

a

3 AUG

1500

285°/12

0

15.0

3.v/Palling

9

8 AUG

1700

270°/1 0

3

114-.3

5.1 /Falling

10

10 AUG

1230

Ol£>°/7

7

111.. 6

3.i>/Rising

11

12 AUG

2130

2l^5°/6

6

Mi. 2

5.5/HHW

12

15 AUG

2230

0

10

16.3

6,9/HHW

13

16 AUG

OltfO

0

8

15.5

-0.7/LLW

11}.

18 AUG

1400

275°/1 3

0

16.0

5.1 /LHW

15

23 AUG

1130

325°/1 0

0

15.8

3.0/HLW

16

25 AUG

114.00

255°/i 0

0

16.0

3,ij./Rising

17

27 AUG

1130

325°/i 2

0

16.5

3.3/Falling

18

30 AUG

H4.00

300°/ii|.

7-

15.3

3.6/LHW

19

1 SEP.

1030

315°/12

0

15.3

2.8/Ri'sing

20

3 SEP

1000

315°/12

10

15.1]-

2.5/Rising

'"'Times represent the approximate time that sampling
■was half completed.

24

TABLE 2

SAMPLING AND INOCULATION TIMES

Survey

Sampling
Commenced
vPDT)

Sampling
Completed
(,PDT)

Samples
Incubated
tPDT)

1

1515

1600

1800

2

'. 13S0

*W5

1600 MPN
1730 MP

3

111-50

1550

1755 MPN
1920 MP

h

1220

.1320

• *\$h£ MPN
1615 MP

5

1330

iI{-30

1600 MPN
1630 MP

6 *

1 91lO

2015

2130

7 .

1350

1Z4J4.0

16I{.5

8

1UU-0

1505

1620

9

1630

1730

1900

10

1210

1315

Ui-50

11

2100

21 W

2250

12.

2215

22i;5

214.00

13

0I|.20

0515

0615

l"fc

13ij-0

1500

1730

15

1100

1200

124.66

16

r%$

1500

1700

17

1100

1200

1l(.00

18

13W"

Ujlj.0

1700

19

1000

1100

1330

20

Ov30

1030 .

1330

25

are from an extraneous source or if they are tongues of
higher colif orm water extending through the surf zone
from deeper water.

The wind during these observations showed little
variation in direction. On all but two of the surveys
along the beach, the winds were westerly with a small
northerly component. On the two excepted^ surveys
(4 and 10), the winds were predominately northerly and
the same general patterns of coliform concentrations were
observed along the beach.

Nearly all tidal situations occurred during these
surveys. Surveys 12 and 13 correspond to extreme levels
of tide of 6.9 and -0.7 feet respectively. Both surveys
were made at night with zero wind velocities. The con-
centration observed in the low tide survey were slightly
larger than those of the high tide survey. A similar
pair of surveys (surveys 5 and 6) were conducted on
14 July with similar weather conditions and contrasting
tidal levels. On this occasion the high tide survey
appeared to have the higher concentration. No consistent
pattern of coliform concentration variation with conditions
of the tide could be determined. This agrees with Steven-
son's |6j observations on the absence of a tidal circulation

in this region.

26

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27

No consistent pattern could be attributed to variation
of sea surface temperature. Sea surface temperature in
the surf zone was found to be quite variable. The anomal-
ously high temperatures that were recorded on surveys 4
and 5 are believed to be due to conduction of heat from
the warm beach to the water in the surf zone. The higher
concentrations of coliforms observed during surveys 11,
12 and 13 may possibly be explained by the absence of the
bacteriocidal ultraviolet radiation from the sun, as these
surveys were taken at night.

To evaluate the effect of storage of samples at room
temperature while awaiting inoculation, increments of 25 ml
of a sample from station 1 were filtered at one hour intervals
for three hours. No significant differences were observed
in the concentrations of coliforms, but non-coliform colonies
did decrease with storage. Since this is within the range of
times that samples were stored during most of the surveys,
changes in concentration during storage were not considered
important.

Large differences were observed between the MPN and MF
methods during the initial surveys with the MPN method giving
concentrations usually an order of magnitude or more higher

28

than those with the MF method (Table 3). Perhaps the use
of five replicates with additional sample volumes would
bring the results in closer agreement, but the additional
time and space required would have further limited the
number of samples that could be analyzed on each survey.
The MPN procedures were suspended after survey 5 in favor
of the more rapid membrane filter method.

Discussion of Surveys 14 through 20

The coliform concentrations determined for the off-
shore surveys were tabulated in Tables 4 through 10. Since
these concentrations were determined by the filtration of
various volumes (50 ml-0.01 ml), the number of significant
figures in the tables represent the number of significant
figures counted. For instance 89 x 104 per 100 ml is
interpreted as 89 colonies were counted from filtration of
0.01 ml.

The data of Tables 4 through 10 were plotted geographi-
cally and iso-concentration lines drawn using a contour
interval of 10 coliforms per 100 ml. Where indeterminate
results were recorded, the order of magnitude of the concen-
tration was estimated on the basis of volumes filtered.
These illustrations are presented as figures 3 through 7.

29

Survey 14 was undertaken primarily to determine the
general extent and concentration of coliforms for use in
planning later offshore surveys. In figure 3, a fairly
tight gradient of concentration lines is found on the
seaward side of the outfall boil with a lesser gradient
towards shore which indicates a general onshore movement
of coliforms and their accompanying surface waters. The
wind for this survey was westerly and the tide is falling.
The seaward extent of water with coliform bacteria is
less than 200 yards from the outfall boil.

Figure 4 represents the data from three surveys.
Survey 15 was conducted along the outside of the surf zone
on 23 August (stations 11 through 22). Survey 16 covered
the next seaward line of stations (stations 23 through 34)
and survey 17 covered stations 35 through 55. The winds
of these surveys were similar in magnitude but the direction
of survey 16' s wind was 255° while the winds of survey 15
and 17 were 325 . Although the value of such a composite
distribution may be questioned, several of the general
features of this distribution may be pointed out. The
pattern of the elongated tongues to the east of the outfall
region were not observed in later more instantaneous surveys,

30

TABLE i*

COLIFORM CONCENTRATIONS DETERMINED ON
SURVEY 11* ON 18 AUGUST 1966

Station Coliform Concentration
(coliforms per 100 ml)

11*- 128 x 10'

26 294 x 10«

35 k

36 0

37 10

38 c

39 c'

/ .

1*0 26

1*3 8

1*6 6

5o 10

60 0

62 0

c

designates indeterminate result because of

overcrowding or massive growth on filters

31

r-i

<o
to

CT5

-P

W

CO

o

•H
«H

O

o

<h

o
a

o

•H

,0

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U

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CQ

o

CO

60
•H

32

but they do indicate the large changes in the field which
occur on a daily or shorter basis. The pattern of iso-
concentration lines to the west are more continuous and
were found to be more representative. Several isolated
pockets of non zero coliform densities were observed at
the western section of the survey region.

Since it was not practical to survey the entire
region on any one survey to obtain a more instanteous
distribution, it was decided to concentrate on the region
around the outfall and survey this reduced region more
completely and in as short a time span as possible.
Figures 5, 6, and 7 are three such surveys which represent
the field based on samples collected within one hour.

The pattern of iso-concentration lines in figure 5
is elongated along a line which is nearly perpendicular
to the wind. Tight gradients are present to the north
and east of the outfall boil.

Figure 6 shows the distribution on 1 September.
Here the wind is more northerly and the coliform distri-
bution is more symmetrical with a greater amount of
spreading to the west. A tongue of higher coliform
water extends from the boil to the southwest toward the
surf zone.

33

TABLE 5

COLIPORM CONCENTRATIONS DETERMINED ON
SURVEY 15 ON 23 AUGUST 1966

Station Coliform Concentration
(coliforms pep 100 ml)

11

12

13
.114.

15
16

17
18

19
20
21
22

:
38

28

1U.0

20

226 x 102

36 x 102

2

2

0

116

26

0

8

260 x 10^

34

TABLE 6

COLIFORM CONCENTRATIONS DETERMINED ON
SURVEY 16 ON 2$ AUGUST 1966

Station Coliform 'Concentration
vcolifomns pop 100 tH)

23

24

25

26

27
28

29
30
31

32
33

38

designates filters on which overcrowding
of non-sheen colonies may influence coliform counts

0

0

238b

209 x

103

0

k

.2

0

0

0

0

0

21 ij. x

10^-

35

TABLE 7

COLIFORM CONCENTRATIONS DETERMINED ON
SURVEY 17 ON 27 AUGUST 1966

Station Coliform Concentrations Station
(coliforras per 100 ml)

Coliform Concentration
vcoliforms' per 100 ml)

35
36
37

38
39
il-0

k2

10

16

10x10'

123 x 10

100

0

2

0

0

32

0

k

I4.6

1

w

It

I|_8

8

&9

ij-

50

134-

* /

2

52

0

53

0

51*

0

55

0

36

o o

o

O-T

95 d
>-J o

9

«
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fa

M

o
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w

00

w
«

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<
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O

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I

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h

'H

TABLE 8.

COLIFORM CONCENTRATIONS DETERMINED ON
SURVEY 18 ON 30 AUGUST 1966

Station Coliform Concentration
(coliforms per 1 00 ml )

Station Coliform Concentratioi
icoliforms per 100 ml.

11

12

13

%

15

16

17

23
2k
25
26

27
28

0

0

0
2 x 103
76
18:
30

0

0

0

k.0 x 10
13 x 10;
10

k

,29

35
36
37
38
39

in

w

W

50

o

2
6
17 x 102
128 x 10^

386b

k

0

2

0
32
52

designates filters on which overcrowding of non-sheen
colonies may influence coliform counts

38

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i

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TABLE y

COLIFORM CONCENTRATIONS DETERMINED
SURVEY 19 ON 1 SEPTEMBER 1966

ON

•

Station

Coliform Concentration
vcoliforms per 100 ml)

Station

Colii
vcoli

'orm Concentration
.forms per 100 ml)

11

30

V

%"

12

i>0

w

0 ||

13

50 x

102

M

k

Ik

360 x

102

w

10

15

130

w

. 0

16

136

50

c

17

3k

51

100

19

0

52

1lj.

35

10

53 /

62

36

16

55

0

37

8

5v

0 .

3Q 8y x 10^ 60 0

3v 2k x 102 61 0

lj.0 10

designates filters on which overcrowding of non-sheen
colonies may influence coliform counts

40

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g

Pd
o

M
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O
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53
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Figure 7 shows the distribution on 3 September. The
pattern presented differs from figure 6 although the
observed wind conditions were nearly identical. Higher
coliform density water has penetrated in all directions.
The only explanation that can be offered for this phenom-
enon is the presence of the dense fog on 3 September which
may have reduced the bacteriocidal effects of solar radia-
tion.

Differentiation of Coliform Bacteria

Completed test for the presence of coliform bacteria
were run on representative coliform cultures obtained during
the sampling process. 106 of these tests were run resulting
in 91 positive tests. Each of these 91 coliform cultures
were submitted to the IMViC biochemical differentiation
tests. 22 or 24.2% of these gave the typical reactions of
Escherichia coli (indole positive, methyl red positive,
Voges-Proskauer negative and citrate negative), while 38
or 41.7% gave the typical reactions of Aerobacter aerogenes
(indole negative, methyl red negative, Voges-Proskauer
positive and citrate positive), and 31 or 34.1% gave
either intermediate or atypical results. It is likely
that the atypical results were caused by cultures which

42

TABLE 10

C0LIF0RM CONCENTRATIONS DETERMINED ON
SURVEY 20 ON 3 SEPTEMBER 1966

Station Coliform Concentration Station Coliform Concentration
v coliforms per 100 ml) icoliforms per 1 00 ml )

11 522b i+0 2 x 102

12 c 1+1 l+10b

13 3^6 x 102 £3 11*.
12+. 107 x 102 i+7 1+

15 17 x 102 2+8 91 0b

16 196 L+9 c

522b

c

31+6 x

102

107 x

102

17 x

102

196

c

c

c

31k-

,

c

81+ .x'

1<A

62+ x

103

17 c 50 21+2 x 10*

19 c 52 ''' 1+2

35 ' c 53 70

36 t 31k- ; 55 ' 56

37 c 59 131+

38 8I|i,x'10^ 60 0b

39 61+ x 1 03 61 1+

designates filters on which overcrowding of non- sheen
colonies may influence coliform counts

c designates indeterminate result because of overcrowding
or massive growth on filters

43

o

o

ft.

M

o
o

EH

125

w

CO

«

CO

§

O
CM

CO
CO
Ci

0)

©
©

CO

to

a

o
as

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o
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o

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o

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iH

o
o

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o

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44

were pure "coliform cultures" but not pure cultures in the
usual sense, since no particular steps were taken to
determine the purity of cultures.

The significance of any particular coliform species
over another has had no observable effect on this study
as there is little doubt of the origin or source of con-
forms depicted around the outfall boil.

45

5. Conclusions

The distribution and concentration of coliform bacteria
have been determined, plotted and qualitatively analyzed for
the surface waters surrounding the outfall of the Monterey
Water Pollution Control Plant. The following general
features of the surface coliform field are concluded:

(1) The general trend of coliform concentrations in
the surf zone along the beach is a region of maximum coli-
form concentration to the east of the outfall with the
level remaining high to the outfall, a rapid decrease of
concentration to the west of the outfall with occasional
isolated centers of higher coliform concentrations, and a
slight increase in concentration towards Monterey Municipal
Wharf No. 2.

(2) In the offshore region a general onshore mass
transport of surface waters in indicated by the rapid decrease
in coliform concentration seaward of the boil and a more
gradual decrease in concentration shoreward of the boil.

This phenomenon was observed in response to westerly and
northwesterly winds.

(3) The seaward extend of coliforms was limited to
several hundred yards north of the outfall boil.

46

(4) The east-west variation of coliform concentration
between the outfall boil and the surf zone was seen to be
quite variable.

Meteorological and oceanographic parameters observed
had the following influences on this general pattern of
coliform distribution:

(1) The tide and water temperatures within the
ranges observed have little or no consistent effect on
the distribution and concentration of coliform bacteria.

(2) The more extensive distribution on four surveys
(surveys 11, 12 , 13, and 20) may in part be attributable
to a decrease in bacteriocidal solar radiation.

(3) The winds observed during this study showed
little variation. No changes in the distribution were
observed in the surf zone in response to small changes
in wind direction or speed. In the offshore region the
pattern of iso-concentration lines tended to be elongated
along a line perpendicular to the wind direction on
surveys 18, 19, and 20.

47

ACKNOWLEDGEMENTS

The author is indebted to Associate Professor E. C.
Haderlie, U. S. Naval Postgraduate School, for guidance
and assistance rendered during all phases of this project.
Appreciation is also expressed to Monterey Peninsula College
and in particular to Dr. W. Trason and Mrs. V. Fry for the
use of the bacteriology laboratory and facilities; to Mr. Al
Hart, supervisor of the Monterey Water Pollution Control
Plant, for advise and use of publications; and to Mr. L. B.
Bowhay, Harbormaster of the City of Monterey, for the use
of the city's boat in the offshore surveys.

48

BIBLIOGRAPHY

1. Breed, R. S., E. G. D. Murray, and N. R. Smith.

Bergey's Manual of Determinative Bacteriology,
7th ed. Williams and Wilkins, 1957.

2. Levine, M. , H. Minette, and R. H. Tanimoto.

"Characteristics and expeditious detection of
bacterial indices of pollution of marine bathing
beaches," Proceedings of the First International
Conference on Waste Disposal in the Marine
Environment, pp 12-28. University of California,
Berkeley: Pergamon Press, 1959.

3. Tibby, R. B. "Inshore circulation patterns and the

oceanic disposal of waste," Proceedings of the
First International Conference on Waste Disposal
in the Marine Environment, pp 296-327. University
of California, Berkeley: Pergamon Press, 1959.

4. Carlucci, A. F., P. V. Scarpino and D. Pramer.

Evaluation of factors affecting survival of
Escherichia coli in sea water. Applied Micro-
biology. V. 9, 1961, 400-404

5. Brennan, J. F., and R. P. Meaux. Observations of

the nearshore water circulation off a sand beach.
M. S. thesis, U. S. Naval Postgraduate School,
Monterey, California, 1964.

6. Stevenson, CD. A study of currents in southern

Monterey Bay. M. S. thesis, U. S. Naval Post-
graduate School, Monterey, California, 1964.

7. Standard Methods for the Examination of Water and

Wastewater, 12th ed. American Public Health
Association, Inc., 1965.

8. Anonomous . Acceptance of membrane filter procedure.

Journal of the American Water Works Association.
V. 50, 1958, 72-74.

49

9. Jannasch, H. W., and G. E. Jones. Bacterial
populations in sea water as determined by
different methods of enumeration. Limnology
and Oceanography, V. 4, 1959, 128-139.

50

INITIAL DISTRIBUTION LIST

No. Copies

1. LT David S. Trumbauer 1

USS Hassayampa (AO 145)

c/o FPO San Francisco, California 96601

2. Professor E. C. Haderlie (Thesis Advisor) 1

Department of Meteorology and Oceanography
U. S. Naval Postgraduate School
Monterey, California 93940

3 . Library 2

U. S. Naval Postgraduate School
Monterey, California 93940

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U. S. Naval Postgraduate School
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5„ Defense Documentation Center 20

Cameron Station
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6. U. S. Naval Oceanographic Office 1

Attn? Division of Oceanography
Washington, D. C. 20390

7. Office of Naval Research 1

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Attn 5 Biology Branch (Code 446)

Washington, Da C. 20360

8. Director 1

Scripps Institution of Oceanography
University of California, San Diego
La Jolla, California

51

9. Chief Scientist

TeVega Expeditions

Hopkins Marine Station

Pacific Grove, California 93950

10. Dr. W. Trason

Department of Biology
Monterey Peninsula College
Monterey, California 93940

11. Mr. Al Hart

Water Pollution Control Plant

City Hall

Monterey, California 93940

12. Mr. Russell White

Monterey County Public Health Department
1200 Aguajito Road
Monterey, California 93940

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Bureau of Sanitary Engineers
2151 Berkeley Way
Berkeley, California 94704

14. Central Coastal Regional Water Pollution
Control Board

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52

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(Security classification ot title, body ot abstract and indexing annotation muat be entered when the overall report la ctaaaified)

1. ORIGINATING ACTIVITY (Corporate author)

Environmental Sciences Programs
U. S. Naval Postgraduate School
Monterey, California

2a. REPORT SECURITY C LASSI FIC ATION

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2b GROUP

3. REPORT TITLE

A COLIFORM BACTERIA SURVEY OF MONTEREY BAY
OFF DEL MONTE BEACH

4. DESCRIPTIVE NOTES (Type ot report and incluaive datea)
THESIS

5 AUTHOR(SHLas(n«me, ti rat name, Initial)

TRUMBAUER, David S.

6- REPORT DATE

October 1966

7a. TOTAL NO. OF PAGES

53

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8a. CONTRACT OR GRANT NO.

b. PROJECT NO.

9a. ORIGINATOR'S REPORT NUMBERfSJ

9b. OTHER REPORT UO(S) (A ny other numbers that may be assigned
this report)

This document has been.apprcvad tetpMOT^

10. AVAILABILITY/LIMITATION NOTICES

release

and sale; its disifioution if unliill

II. SUPPLEMENTARY NOTES

12- SPONSORING MILITARY ACTIVITY

U. S. Naval Oceanographic Office
Washington, D. C. 20390

13. ABSTRACT

Determination of the distribution of coliform bacteria
in the vicinity of the outfall effluent of the Monterey Water
Pollution Control Plant was made using the membrane filter tech-
nique for enumeration. The coliforms were seen to follow the
onshore mass transport of water in the surface layer in response
to predominately westerly winds. The extent of penetration of
coliforms into the bay north of the outfall is restricted to
several hundred yards, while more extensive spreading was observed
towards the surf zone. No consistent pattern of variation of the
coliform distribution would be related to the tidal cycle. Higher
concentrations of coliform bacteria on several occasions seemed
to be related to reduced periods of solar radiation..

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COLIFORM BACTERIA

MONTEREY BAY

MEMBRANE FILTER

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