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ENUMERATION
OF MARINE BACTERIA>
John D. Buck and Robert C. Cleverdon
Department
of Bacteriology,
University
of Connecticut,
Storrs, Connecticut
ABSTRACT
A comparison was made of agar plates spread with glass rods and poured agar plates for
the enumeration
of bacteria in the waters of Fishers Island Sound, salinity 30&. Spread
plates were shown to he markedly superior.
Highest counts were obtained by spreading,
using rods treated with Desicotc (a silicone solution) and incubating plates at 25C, rather
than at 16C, and in air rather than in air with CO2 content increased.
INTBODUCIION
AND
MATERIALS
Water was obtained at various tidal conditions during the summer and fall of 1958,
from one location at Latimer Reef, which is
about 6 miles off the coast of Noank, Conneticut, and east of East Point, Fishers
Island, New York. Top samples were collected in sterile 500 ml wide-mouth reagent
bottles. Samples were iced until examination at the laboratory, within 2 hours.
Preliminary
studies with samples from
Fishers Island Sound showed that counts
could be obtained using decimal dilutions
of 10-l to lo-. Dilution blanks (9 and 99
ml of sea water) were sterilized in the autoclave. Pipettes were 1.1 ml (milk) pipettes,
sterilized in the oven, and stored in cans.
1 Supported in part by Grand #E-706,
Institutes of Health.
2 Contribution
#l from the Marinc
Laboratory,
University
of Connecticut,
Connecticut.
National
Research
Noank,
ENUMERATION
TABLE 1.
nntc
EllKl
saE-plc
.
Tide
OF MARINE
BACTERIA
BY THE
SPREAD
79
PLATE
Ratios of counts
RiYs
Spread
Pour
Ratio
S/P
1.1
0.71
S air/S CO2
P air/P CO2
7/17
Flood
26
14
1.8
23
20
1.2
7/25
Ebb
53
12
4.4
41
23
1.8
1.3
0.52
7/31
Ebb
11
5.5
19
9.5
0.58
1.0
817
Low
140
77
1.8
86
51
1.7
1.6
1.5
8/14
High
11
3.7
1.2
1.6
0.5
8/21
Flood
150
64
2.4
130
99
1.3
1.2
0.65
8/28
High
4.0
11
3.7
0.73
0.67
9/4
Flood
35
12
2.9
41
31
1.3
0.86
0.39
9/10
High
1.6
2.5
1.6
2.5
10
9/19
Ebb
190
120
1.6
110
100
1.1
1.7
1.2
Average ratios
2.9
2.5
1.2
0.96
AND
DISCUSSION
80
JOHN
TABLE 2.
Comparison
Saz?e
Tide
and
Air
AND
ROBERT
C. CLEVERDON
16%
Date
D. BUCK
co2
PQ
Air
D/P
CO2 D
D/P Ai;/
P
g;/
2
25C
-
Air
DP
Desicoted
CO2
DP
CO2 D
D/PAir/
co2
Ratio of counts
25C/l&C
PAir /
co2
D
Air
D
CO2
h%
Co2
14
;W,:
13
1.6
1.2
1.9
1.3
20
12
12
10
1.7
1.2
1.7
1.2
1.5
1.7
1.5
1.7
15
11/l
IIig?
12
1.3
1.3
1.5
1.5
18
18
15
1.0
0.47 2.6
1.2
1.5
0.88 2.0
2.5
16
Et?
190
0.79
1.3
0.63
1.0
0.67
1.1
1.8
2.9
2.5
0.87 2.9
1.0
17
%J5
46
16
1.6
0.56 5.1
1.8
1.4
1.0
3.4
2.5
1.0
1.6
0.88
18
E9
1.2
1.3
3.3
3.9
4.5
1.3
1.1
2.5
1.9
1.1
2.5
1.4
1300
35
14
14
0.88 3.1
1.1
1.2
1.7
1.3
APHA,
AWWA
AND FSIWA.
1955. Standard
methods for the examination of water, sewage,
Tenth ed. APHA, Inc.
and industrial wastes.
New York.
522 pp.
CARLUCCI, A.F., AND PRAMER, D.
1957. Factors
influencing
the plate method for determining
Proc.
abundance
of bacteria in sea water.
Sot. Exptl. Biol. Med., 96: 392-394.
ZOBELL, C. E. 1948. Marine microbiology.
Chronican Botanica Co., Waltham Mass. 240 pp.
ZOBELL, C. E. 1954. Bacteriology
of the sea. pp.
5@3-Sl6. In Salle, A. J., Fundamental
princiMcGraw-Hill
Book Co.,
ples of bacteriology.
Inc. New York.
782 pp.