PHYSICAL, CHEMICAL, AND BIOLOGICAL
OCEANOGRAPHIC OBSERVATIONS
OBTAINED ON EXPEDITION SCOPE
IN THE EASTERN TROPICAL PACIFIC
NOVEMBER - DECEMBER 1956
SPECIAL SCIENTIFIC REPORT-FISHERIES No. 279
UNITED STATES DEPARTMENT OF THE INTERIOR
FISN AND WILDLIFE SERVICE
EXPLANATORY NOTE
The series embodies results of investigations, usually of restricted
scope, intended to aid or direct management or utilization practices and as
guides for administrative or legislative action. It is issued in limited quantities
for official use of Federal, State or cooperating agencies and in processed form
for economy and to avoid delay in publication .
UNITED STATES DEPARTMENT OF THE INTERIOR, Fred A. Seaton, Secretary
Fish and Wildlife Service, Arnie J. Suomela, Commissioner
PHYSICAL, CHEMICAL, AND BIOLOGICAL OCEANOGRAPHIC OBSERVATIONS
OBTAINED ON EXPEDITION SCOPE IN THE EASTERN TROPICAL PACIFIC
NOVEMBER - DECEMBER 1956
By Robert W. Holmes and other members of the
Scripps Cooperative Oceanic Productivity Expedition
Part 1. Methods and Station Data.
Part 2. Scientific Reports.
"This work was financed by the Bureau of
Commercial Fisheries under Contract No.
14-19-008-2485, with funds made available
under the Act of July 1, 1954 (68 Stat.
376), commonly known as the Saltonstall-
Kennedy Act."
Special Scientific Report — Fisheries No. 279
Washington, D. C.
November 1958
The Library of Congress has cataloged this publication
as follows:
Scripps Cooperative Oceanic Productivity Expedition, 1950.
Physical, chemical, and biological oceunogruphic observa-
tions obtained on Expedition SCOPE in the eastern tropical
Pacific, November-December 1956, by Robert W. Holmes
and other members of the Scripps Cooperative Oceanic Pro-
ductivitv Expedition. Washington, U. S. Dept. of the
Interior", Fish and Wildlife Service, 1958.
117 p. ump, dinars. , tables. 27 cm. (U. S. Fish and Wildlife
Service. Special scientific report : fisheries, no. 279)
Includes bibliographies.
1. Pacific Ocean. l. Holmes, Robert W. n. California. Uni-
versity. Scripps Institution of Oceanography, La Jolla. (Series*
SH11.A885 no. 279 551.466 59-60425
Library of Congress
The Fish and Wildlife Service series, Special Scientific
Report — Fisheries, is cataloged as follows:
U. S. Fish and Wildlife Service.
Special scientific report : fisheries, no. 1-
iWashingtoii] 1949-
no. illus., maps, diagrs. 27 cm.
Supersedes in part the Service's Special scientific report.
1. Fisheries — Research.
SH11.A335 639.2072 59-60217
Library of Congress (2j
ABSTRACT
This SCOPE report describes the methods employed, lists in
tabular form the results obtained, and includes a series of papers which
discuss the results of a preliminary analysis of certain of the biological
observations which were obtained on a cruise to the Eastern Tropical
Pacific accomplished by the University of California, Scripps Institution
of Oceanography, under Contract No. 1^-19-008-2^5 with the Department of
Interior, U. S. Fish and Wildlife Service. Scientific personnel, equipment,
and financial support for the data analysis have been largely provided by the
University of California, Scripps Institution of Oceanography and the Inter-
American Tropical Tuna Commission.
Information was obtained on the vertical and horizontal
variations in temperature, salinity, dissolved oxygen, inorganic phosphorus,
nitrite, alkalinity, pH, chlorophyll "a", primary production, bacterial
abundance, and zooplankton standing crop. A nearly continuous record of
incident solar radiation was obtained and was accompanied by daily measure-
ments of the attenuation of blue-green light in the ocean. Water samples
and fine-mesh net-hauls were collected for the subsequent analysis of
phytoplankton abundance and species composition. The distribution of
vertebrates was also studied with special emphasis on oceanic bird distri-
but ion .
Nine scientific papers which are the result of an analysis of
certain of the SCOPE data are included in Part 2 of this report. They are:
Possible application of a bacterial bioassay in productivity studies, by
William Belser; SCOPE measurements of productivity, chlorophyll "a,r7 and
zooplankton volumes, by R. W. Holmes, M. B. Schaefer, and B. M. Shimada;
Size fractionation of phot o sy nt he s i z i ng phytoplankton, by Robert W. Holmes;
Diurnal variation in the photosynthesis of natural phytoplankton populations
in artificial light, by Robert W. Holmes and Francis T. Haxo; Attachment of
marine bacteria to zooplankton, by Galen E. Jones; Preliminary studies of
bacterial growth in relation to dark and light fixation of Cl%02 during
productivity determinations, by G. E. Jones, W. H. Thomas, and F. T. Haxo;
The effects of organic and inorganic micronutrients on the assimilation of
Cl^ by planktonic communities and on bacterial multiplication in tropical
Pacific sea water, by Galen E. Jones and William H. Thomas; The vertebrates
of SCOPE, November 7 - December l6, 19^6, by Robert Cushman Murphy; The
alcohol- soluble and insoluble fractions of the photosynthetically fixed
carbon in natura% occurring marine phytoplankton populations, by "
William H. Thomas.
CONTENTS
Page
PART 1 . METHODS AND STATION DATA, by Robert Holmes 1
Introduction 3
Procedure at noon stations 4
Procedure at in situ productivity stations 4
Procedure between stations 4
Continuous observations .
Methods 4
Incident solar radiation 4
Submarine daylight 5
Salinity, temperature, depth 5
Surface current by Geomagnetic Electrokinetograph (GEK) . 7
PH 7
Alkalinity 7
Nitrite 7
Inorganic phosphorus 7
Dissolved oxygen 7
Chlorophyll "a"
Primary production 3
Zooplankton standing crop g
Bacteria 9
Noon station data i-.
Observations between noon stations 45
GEK observations c-i
Page
PART 2. SCIENTIFIC REPORTS 53
Possible application of a bacterial bioassay in
productivity studies.
William Belser 55
SCOPE measurements of productivity, chlorophyll
"a", and zooplankton volumes.
R. W. Holmes, M. B. Schaefer, and
B. M. Shimada 59
Size fractionation of photosynthesizing
phytoplankton .
Robert W. Holmes 69
Diurnal variation in the photosynthesis of
natural phytoplankton populations in
artificial light.
Robert W. Holmes and Francis T. Haxo 73
Attachment of marine bacteria to zooplankton.
Galen E. Jones 77
Preliminary studies of bacterial growth in
relation to dark and light fixation of
Cl*K)2 during productivity determinations.
G. E. Jones, W. H. Thomas, and F. T. Haxo ... 79
The effects of organic and inorganic
micronutrients on the assimilation of C^ by
planktonic communities and on bacterial
multiplication in tropical Pacific sea water.
Galen E. Jones and William H. Thomas 87
The vertebrates of SCOPE, November 7 -
December l6, 1956.
Robert Cushman Murphy 101
The alcohol- soluble and insoluble fractions of
the photosynthetically fixed carbon in
naturally occurring marine phytoplankton
populat ions .
William H. Thomas 113
PART 1. METHODS AND STATION DATA
By
Robert W. Holmes
- 1 -
INTRODUCTION
SCOPE (Scripps Cooperative Oceanic Productiv-
ity Expedition) was a cooperative biological
oceanographic survey of the eastern tropical
Pacific conducted during November and December
1956. The expedition was designed to examine
regional variations in primary production in
tropical areas of interest to the American
tuna fisheries and to provide a basis for
subsequent studies to explain the biological
effects of variations in the oceanic circula-
tion and the influences thereof on the dis-
tribution and behavior of the tunas. Partic-
ipating in this endeavor were scientists from
the Scripps Institution of Oceanography, the
Inter-American Tropical Tuna Commission, and
the American Museum of Natural History. This
study was made possible by the establishment
of a contract (No. 1^-19-008-2^5) between the
University of California, Scripps Institution
of Oceanography and the Department of Interior,
U. S. Fish and Wildlife Service. Personnel
and additional financial support were provided
by the University of California, Scripps Insti-
tution of Oceanography and the Inter-American
Tropical Tuna Commission.
In addition to fulfilling the objectives list-
ed above, some effort was devoted to a study
of the biological methods employed and to a
study of certain fundamental biological problems
which have a bearing on the productivity of
ocean waters. Thus;. certain studies on the
standing crop and nutritional requirements of
bacteria, on the solubility of phytoplankton
protoplasm, and on the distribution of organic
growth factors were included in the observa-
tional program. Studies of this nature will
eventually contribute to a better understanding
of events at low trophic levels in the food
chain and help us understand the interchange
and interaction between the chemical environ-
ment, the phytoplankton and phytoplankton
production.
The expedition departed from San Diego on the
M/V Stranger on November 7th, 1956 and returned
to San Diego on December 17th, 1956. The track
is illustrated in Figure 1.
The following is a list of scientific per-
sonnel participating in the expedition:
Robert W. Holmes, expedition leader,
Assistant Research Biologist, Scripps Insti-
tution of Oceanography, University of Calif-
ornia
Dr. William H. Brandhorst, Scientist,
Inter-American Tropical Tuna Commission
Dr. Francis T. Haxo, Assistant Professor,
Scripps Institution of Oceanography, Univer-
sity of California *
Dr. Galen E. Jones, Assistant Research
Biologist, Scripps Institution of Oceanog-
raphy, University of California
Robert J. Linn, Senior Marine Technician,
Scripps Institution of Oceanography, Univer-
sity of California
Dr. Robert C. Murphy, Lamont Curator
Emeritus of Birds, American Museum of Natural
History
Park Richardson, Laboratory Technician,
Scripps Institution of Oceanography, Univer-
sity of California
Dr. Milner B. Schaefer, Director, Inter-
American Tropical Tuna Commission
Dr. Bell M. Shimada, Senior Scientist,
Inter-American Tropical Tuna Commission
Dr. William H. Thomas, Assistant Research
Biologist, Scripps Institution of Oceanog-
raphy, University of California**
* Panama to San Diego
** San Diego to Panama
Not all of the data and material collected
on the expedition have been analyzed. The
phytoplankton standing-crop samples have not
been examined nor have many of the possible
interrelationships between the biological,
chemical and physical observations been studi-
ed. This work is presently being carried out
by Robert W. Holmes. The information on the
distribution of oceanic birds is presently
being incorporated into a monograph on trop-
ical oceanic birds by Dr. Robert Cushman Murphy.
Certain of the data obtained on the expedition,
and included in this report, have been
presented at scientific meetings. Drs.
G. Jones and W. Belser presented papers at
the Detroit, Michigan, meetings of the Society
of American Bacteriologists in April-May 1957
which included information obtained on SCOPE.
PROCEDURES AT NOON STATIONS
At approximately local noon of each day,
weather permitting, a station was occupied.
The general procedure at these stations was
as follows:
1. 900 ft. - BT lowering and general
weather observations including barome-
ter reading, dry- and wet-bulb air temp-
eratures, wind direction and speed, sea
and swell observations, and sky condition.
2. Collection of surface water sample for
trailing bottle productivity studies.
3. Submarine photometer lowering.
h. 50 m. - Surface, vertical phytoplankton
net haul using a i+O-cm. truncate net,
with a mesh size of 32^.
5. Plastic sampler cast to 100 m. - Water
samples collected were used in photo-
synthetic studies in the shipboard
incubator and for the determination
of chlorophyll "a" concentrations. A
small aliquot from each depth was
also preserved for subsequent phytoplank-
ton analysis.
6. J-Z sampler cast for bacterial abundance
studies.
7- Nansen bottle cast to approximately 700 m.-
The water samples were employed for oxygen,
salinity, alkalinity, inorganic phosphorus,
pH, and nitrite determinations.
8. Oblique zooplankton meter-net tow to a
depth of approximately 300 m.
PROCEDURE AT IN SITU PRODUCTIVITY STATIONS
(S-9 SERIES, S-20, S-25A, and S-25B)
Shortly after arrival at these stations a
surface parachute drogue was released and
all subsequent observations were taken along-
side the drogue.
The sampling program was rather variable but
consisted of a series of observations, casts,
etc., similar to those taken at each noon
station. At the S-9 stations several hydro-
graphic casts were made with the Nansen
bottles very closely spaced.
The area in which the S-9 station series
were located is referred to in this report
as "the Dome" or as the thermal anticline
region. This is a large region lying off
the west coast of Costa Rica characterized by
an intense, shallow thermocline. This is
an area of high productivity in which the
characteristics of upwelled water are absent.
. PROCEDURES BETWEEN STATIONS
While underway, between noon stations, 900-ft .
BT lowerings were made every three hours
(0000, 0300, 0600, 0900, 1200, 1500, l800,
and 2100 hours) accompanied by routine
weather observations. Surface chlorophyll
"a" and inorganic phosphorus determinations
were frequently made at 0600 which was also the
usual time that the morning trailing bottle
productivity experiment began.
CONTINUOUS OBSERVATIONS
1. Sea-surface temperature was continuously
recorded with a Taylor thermograph.
2. Incident solar radiation was measured
by a 10-junction Eppley pyrheliometer
combined with a Speedomax 0-10 mv recorder.
METHODS
1. Incident solar radiation:
A gimbals-mounted Eppley 10-junction
pyrheliometer was placed above all super-
structure on the afterma6t of the m/v
Stranger . The signal from the pyrheliometer
was fed into a 0-10 mv Speedomax recorder
and was recorded on chart paper travelling at
the rate of two in. per hour.
The integration yielding the daily radiation
total was performed with a planimeter and day
length was computed from the Speedomax trace.
The value for the daily total is given, together
with other data, in the tables containing the
noon station observations.
2. Submarine daylight
Two different filters and submarine photometers
were utilized in the measurement of submarine
daylight. The transmission characteristics of
the two filters (Chance OB- 10 and Wratten No.
V?) are given in Figure 2. Since the trans-
mission of the Wratten 1+5 changed somewhat
during its use, a third curve is given which
was made with this filter immediately after the
return of the expedition. The darkening of
the 1+5 filter was assumed to be a gradual process
and, as there was no significant shift in the
spectral characteristics, all readings made
with this filter are believed to be comparable.
Both of the photometers employed were essentially
identical, the only important difference being
that the collector plate (abraded translucent
plastic) in the first instrument (used from
Station S-l to S-1+) was elevated above the
instrument housing in such a manner that the
flat plate collector was not shadowed and had
an angle of acceptance of IflO" . In the second
instrument, used throughout the remainder of
the expedition, the angle of acceptance of
the collector was somewhat less than l8o° owing
to the fact that a shoulder on the photometer
housing rose a few millimeters above and around
the collector plate.
With a single exception reducing screens were
not employed. The photometer was lowered in the
water until the output of the Photronic cell
(Weston 856, Type RR) in the photometer was
less than 1000 ua (usually at about 2 m. depth) .
The output of a gimbals-mounted deck cell, like-
wise filtered with a Wratten 1+5 filter, was not-
ed at the same moment as the output of the sub-
merged cell was recorded. This process was
repeated at successive depths until the output
of the submarine cell was too low to be
measured with the microammeter . Due to
fluctuations in ambient light and distur-
bance caused by waves and swell, readings at
various depths were not made until an
apparent equilibrium had been reached. In
cases where wave action was particularly
disturbing and/or fluctuations in ambient
light were very marked, simultaneous readings
of the output of the deck and underwater cell
were repeated and an average of these values
was used.
The current generated by the Photronic cell in
the photometer was measured with a damped
Rawson multimeter (0-50, 0-100, 0-200, 0-500,
0-1000 (ia) which possessed an internal
resistance (on all scales) of 100 ohms. The
output of the deck cell was measured with a
0-1 milliampere meter (internal resistance:
50 ohms ) .
The data presented in the tables have been
corrected in the following manner: all
of the submarine photometric data (^a) have
been corrected for departures in linearity of
response of the photronic cell, and these
values in turn adjusted to a constant but
arbitrary incident radiation value (deck
cell reading) . The "true" instrument depth
has likewise been computed from wire-angle
measurements and length of wire out.
The diffuse attenuation coefficient per unit
distance (meter), k, and the percent trans-
mission per unit distance (meter), T, have
been calculated for each depth interval using
the following formulas:
In I
AZ,
- In I
AZ,
- Z,
where I is the corrected output of the
submerged cell, Z is the depth in meters,
and refers to the spectral sensitivity of
the Photronic cell-color filter combination
employed (see Figure 2).
3. Salinity, temperature, depth
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-6 -
Two or more chlorinity determinations were
made with each sample, employing the Knudsen
method, and these were converted to salinity.
Temperature was measured with standard revers-
ing thermometers, and the necessary correc-
tions (index, etc.) were carried out at the
Scripps Institution of Oceanography to give
in situ values. When corrected temperatures
of paired protected thermometers differed by
more than 0.06°C, both values appear in the
tables. The temperatures listed at 0 depth
are actually at an average depth of 1.5 m.
below the sea surface.
Depths are based on readings of paired
(protected and unprotected) reversing ther-
mometers .
Nansen bottle spacing was determined by the
thermal structure of the water and an attempt
was made to place the bottles at equal -temper-
ature intervals rather than at equal-depth
intervals.
Corrections have been made using the tables
of Harvey (The Chemistry and Fertility of
Sea Waters, Cambridge Univ. Press, 224 pp.,
1955).
6. Alkalinity
Alkalinity was determined using the method
of Anderson and Robinson (industrial and
Engineering Chemistry, Analytical Edition,
Vol. 18, p. 767, 19h6). These are reported
for atmospheric pressure and are accurate to
0.01 millival/l.
7-
Nitrite
Nitrite measurements were made with a Beck-
man DU Spectrophotometer employing the
method of Bendschneider and Robinson (Jour.
Mar. Res., Vol. 11, p. 87, 1952). The data
are reported in u gm at/l NO -N, and the
accuracy ranges from 5 to 10%.
8. Inorganic phosphorus
Only values at observed depths appear . As it
will be some time before all station curves
are drawn, it was decided to submit the data
in the present form.
.h. Surface current by Geomagnetic Electro-
kinetograph (GEK)
All measurements were made with neutrally
buoyant cable. The conversion from measured
electrical potential to surface current was by
the formula E = -VH S where E is the measured
potential, V the surface current, H the
vertical component of the earth's magnetic
field, and S the interelectrode distance. No
corrections, therefore, have been made for
"depth of current," "electrode droop," or
"windage on electrodes."
5- pH
The pH of samples was determined with a Beckman
G pH meter employing glass and calomel
electrodes. The values given are for in situ
conditions and are accurate to 0.02 pH units.
Phosphate concentrations were measured
using the method of Wooster (Jour. Mar. Res.j
Vol. 10, pp. 91-100, 1951). Duplicate
samples were not analyzed.
9. Dissolved oxygen
Dissolved oxygen measurements were made
using the Winkler technique according to
the directions of Wooster (Methods in
chemical oceanography. . .employed in the
California Cooperative Sardine Research
Program. Scripps Inst. Oceanogr., Tech.
Rept., 27 pp.).
10. Chlorophyll "a"
The water samples used for the determina-
tion of chlorophyll "a" content were
collected from the surface with a plastic
bucket; subsurface samples were collected
with a Van Dorn-type plastic sampler. The
water sample, 3. 0-6.0 1. in volume, was
shaken after the addition of a small amount
of magnesium carbonate, and filtered through
a 1+7 -mm. type HA, plain, white Millipore
filter. The filter membranes were dried in a
vacuum desiccator and then extracted with 3
mis. of 90°/ acetone (glass redistilled) in
the cold (ca. 10°C) and dark for approximately
10-12 hours. The sample was then centrifuged
until clear. The supernatant was next decanted
into a volumetric flask or cylinder and the
remaining precipitate in the tube resuspended
with 1-2 mis. of 90% acetone, centrifuged,
and the supernatant combined with that obtain-
ed previously. Recentrifugation of the
combined extracts was frequently necessary to
reduce turbidity. This extract was finally
diluted to 6 ml., and its optical density was
measured in a 10-cm. semimicro-absorption cell
at 750, 665, 61+5, and 630 mu with a Beckman
model DU spectrophotometer. Turbidity correc-
tions were made on the basis of the sample
transmission at 750 mu and the concentrations
of chlorophyll "a" have been calculated from
the equations of Richards with Thompson (Jour.
Mar. Res., Vol. 11, No. 2, pp. 156-172, 1952).
11. Primary production
lit-
The C method was employed in these studies
to determine the rate of carbon fixation by
the phytoplankton. The C1^ solution was
prepared and standardized in the manner
described by Steemann Nielsen ('Jour, du Cons.,
Vol. 18, No. 2, pp. 117-l1+0, 1952) with the
exception that glass redistilled water rather
than artificial sea water was used as the
solvent. The C^ solution employed was filter-
ed through an HA Millipore filter and put in
1-ml. glass ampules which were autoclaved.
The radioactivity of the samples was measured
with an NMC-PC#1 proportional counter.
In situ surface productivity was measured
using samples dipped from the sea surface with
a plastic bucket at either sunrise or local
noon. The samples were placed in clean, well
aged, 250-ml. Pyrex bottles inoculated with
C^ , and trailed astern of the vessel, just
under or on the top of the sea surface, until
local noon or sunset, respectively. The
samples were filtered immediately and placed
in a vacuum desiccator for drying.
The in situ vertical measurements of productiv-
ity were carried out in the following manner.
A water sample was collected at each desired
depth with the plastic Van Dorn-type sampler
shortly before daylight. The samples were
transferred to clean, well aged, 250-ml.
Pyrex bottles and the C-1-^ solution injected
with a plastic hypodermic syringe and stain-
less steel needle. The samples were re-
suspended at or slightly before dawn, at
approximately the depth ( + 1 m.) at which
they were collected, on a weighted rope
supported by a free-floating glass buoy (ll+
in. in diameter) enclosed in a cord netting
and attached to a bamboo pole bearing a flag
at its top. The surface sample was attached
to the side of the glass buoy, just under
the sea surface. The samples were collected
at noon, local time, and were promptly filter-
ed and dried for counting.
The samples incubated on shipboard were
inoculated with C in the same manner as the
in situ and trailing bottle material. The
incubator itself was similar to that employed
by Steemann Nielsen (Jour, du Cons., Vol. l8,
No. 2, pp. 117-llK), 1952). Temperature con-
trol was achieved by circulating subsurface
sea water through the water bath at a rate of
It— 6 1 . per minute . The temperature in the
bath fluctuated somewhat but never exceeded
the sea-surface temperature by more than
2.3°C, and usually by less than 1°C.
Temperatures less than that of the sea sur-
face were not observed in the incubator . The
samples were illuminated by a bank of 10
daylight-type fluorescent lamps. The lamp
bank was moveable and was the means employed
in keeping the intensity of light at the
bottles at 1000 foot-candles.
The data presented in these pages have not
been corrected for dark-bottle uptake, the
isotope effect, or for phytoplankton
respiration. In our experience the dark-
bottle uptake averages 10-13*% of the uptake
in the illuminated bottles when the experi-
mental period does not exceed eight hours,
although dark uptake may exceed this if the
bottles are not washed carefully. This
value of 10-13% > which must be substracted from
light-bottle uptake, is nearly equal to the
10°/o positive correction suggested by Steemann
Nielsen (1952). The data have not been correct-
ed for phytoplankton respiration losses during
the hours of darkness. The total CO2 concentra-
tion of sea water has been assumed to equal 90
mg/l. and all of the productivity calculations
have been made using this value.
12.
Zooplankton standing crop
Measurements were made of the standing crop of
zooplankton by means of plankton net hauls,
using gear and techniques comparable to those
presently employed by the California Cooperative
Oceanic Fisheries Investigations. At each station
an oblique tow was made with a one-m. (mouth
diameter) plankton net made of 30XXX silk grit
gauze in the body and 56XXX silk grit gauze in
the rear section and cod-end bag. The net was
lowered from the surface to a depth of approxi-
mately 300 m. (1+50 m. wire length) at a rate of
50 m. per minute while the vessel was slowly
underway and retrieved at a rate of 20 m. per
minute. The duration of a single haul, there-
fore, was about 32 minutes, on the average. An
Atlas flow meter was mounted in the mouth of the
net to record the volume of sea water filtered
by the net. Flow meters were calibrated before
and after the cruise.
Zooplankton collections were preserved in k" / 0
buffered formalin. Ashore, the collections
were filtered and the total "wet" volumes
of plankton obtained at each station were
measured by displacement. The volume of water
sampled by each haul was determined by a
method described by the South Pacific Fishery
Investigations of the U. S. Fish and Wildlife
Service and the displacement volumes were then
converted into terms of the volume of organisms,
in cu. cm., collected from each 1000 cu. m. of
sea water strained.
13 . Techniques used in the abundance determina-
tion of heterotrophic micro-organisms (bac-
teria)
Sea-water samples were collected from various
depths in the water column with sterile
rubber bulbs attached to J-Z water samplers
(ZoBell, Marine Microbiology: A monograph on
hydrobacteriology, Chronica Botanica, 19^) •
The contents of the J-Z samplers were trans-
ferred to sterile 200-ml. prescription
bottles immediately after arriving at the
surface. The bacterial counts were determin-
ed by plating 0.1- to 5-0-ml. aliquots of the
water samples in duplicate in sterile,
plastic, disposable, petri dishes (Falcon
Plastics, Culver City, California). The
medium had the following composition: peptone
(Difco), 5.0 g; yeast extract (Difco), 1.0 g;
FeoPOi,., trace; agar, 15.0 g; aged sea water
(75°/o), 1,000 ml. as defined by Oppenheimer
and ZoBell (The growth and variability of
sixty-three species of marine bacteria as
influenced by hydrostatic pressure, Jour.
Mar. Res., Vol. 11, No. 1, pp. 10-18, 1952).
The sterile agar medium was cooled to ^2°C
t 2°C and was poured into the seeded plates
on a table suspended from the ceiling of the
lounge (below decks). The suspended table
was weighted underneath to provide stability
and steadied with the aid of a second person.
Such a free-swinging table proved sufficient
to compensate for the roll of the ship in calm-
to-moderate seas. The plates were incubated
at 31°C t 1°C, for three days or longer
before reading on a Quebec colony counter.
The high temperature of incubation employed
is not customary for marine bacteria. This
temperature was the lowest possible aboard
ship in the tropics without a refrigerated
incubator. This temperature was not too
high for surface forms since surface sea-
water temperatures were almost as high.
The results of the bacterial counts taken with
the J-Z samples on the return trip from
Panama are reported in the noon station data
tables.
On the cruise from San Diego to Panama bacte-
rial counts were made from water collected
in plastic Van Dorn samplers. This sampler
was used since Wood (Heterotrophic bacteria
in the marine enviroment of eastern Australia.
Australian Jour. Mar. and Freshwater Res.,
Vol. k, No. 1, pp. 160-200, 1953) reported non-
sterile Nansen bottles produced almost the same
counts as sterile samples and since other
determinations were made from these same water
samples. By the end of the trip to Panama it
was evident that the bacterial counts in water
collected in the plastic samplers were much
higher than those obtained in sterile J-Z
samplers. Several direct comparisons were
made at the same place, depth, and time and
10^ to 10^ more cells were taken from the
plastic samplers. The resulting contamination
in plastic samplers apparently developed from
a bacterial film on the sides of samplers due
to their constant use. This was concluded
after observing the gradual increase in bac-
terial numbers after each use. These figures
are not presented.
- 10
NOON STATION DATA
Station 1
M/V Stranger; SCOPE; November 10, 1956; 2050, 21101 GOT; 22°57.0*N, 113'
3l+.5!W; lbOO fm; vire angle, 0°, 0°; temp., 76.0°F dry, 72.0°F wet;
weather, 02; clouds, 6, amt., 8; sea, 2; swell, 330°, 3 ft, 8 sec.
Depth
(m)
0
30
1+0
55
65
109
138
202
290
389
735
T
CO
21+.22
2k. 16
23.90
17.32
13.61
11.96
11.35
10.68
9.1+6
8Jt6
7.3^
5.51
s
C/..)
31+.1+0
31+.1+0
3^.39
3^.1^
33.82
33.31
3I+.1+2
3^.58
3^.56
3^-52
3^.1+9
3^. ^9
(n5/i)
k.k2
^.39
k.kl
3.36
3.31
1.01
0.99
0.37
0.13
0.16
0.10
0.ll+
OBSERVED
PO^-P
(ugm at/l)
0.62
0.56
0.62
1.29
1.39
2.3^
2.1*8
2.96
2.97
3.03
3.18
NO^-N
(ugm at/l)
Alk
pH
(millival/l)
8.19
2.3^
8.21
2.3U
8.09
2.33
8.06
2.32
7.82
2.34
7.82
2.3I+
7.76
2.36
7-73
2.37
7.70
2.37
7-73
2.1*0
7.73
2.1*1
Depth
(■)
0
25
50
100
Chlorophyll
(mg /v?)
.125
.21+5
• 33^
.108
BIOLOGICAL OBSERVATIONS
Productivity-
Bacteria
( no/ml )
in situ incubator B-
(mg c/nr/day) mg c/nr/br
U-
Zooplahkton Volume: 1+9 ml/lOOO nr total, 1+9 ml/lOOO m^ small.
Incident Radiation _
Daily Maximum: 0.622 cal/cm /min.
Daily Total: ll6 cal/cm .
Day Length: 10.32 hrs.
^All the times given in the station headings are messenger time(s).
fBiotin: for explanation of symbols see p. 56 , footnote No. 3.
^Uracil:
Purine: (see p. 51+ ) :
- 11
Station 2
M/V Stranger; SCOPE; November 11, 1956; 2105, 2120 GCT; 21°07.0'N, 110°03.0'W;
1700 fin; wire angle, 3°, 5°; temp., 85.0°F dry, 78.0°F wet; weather, 02;
clouds, 2, amt., 2; sea, missing; swell, confused.
OBSERVED
Depth
Temp.
S
Oo
(ml/D
P01+-P
N02
-N
Alk
(a)
(°c)
%)
(u gm at/l)
(++ gm
a1
;/D
PH
(millival/l)
0
28.12
3I+.62
J+.J+9
_
0.0
8.20
2.38
15
27.65
3^.63
1+.30
0.48
0.0
8.20
2.36
35
27.22
3I+.6O
M5
0.55
tr.
8.20
2.36
50
26.17
3^-51
1+.27
1.13
0.5
8.19
2.36
65
20.96
3^^5
2.61+
1.50
0.1
8.014-
2.35
85
17.72
3^.39
1.93
1.76
tr.
7.96
2.3U
118
13.36
3J4-.ll
2.08
1.86
0.0
7.92
2.33
196
12.26
3^.78
-
2.58
0.0
7.73
2.36
2 1+3
11.76
11 A3
3^.76
-
2.55
0.0
7.77
2.36
290
10.66
3I+.70
0.06
2.66
0.0
7.73
2.37
388
8.89
3^.58
0.25
2.71
0.0
7.71
2.37
1+82
7-7^
3^-52
0.13
2.78
-
7.72
2.37
73^
5.52
3^.53
0.1+1+
2.70
0.0
7.73
2.1+1
BIOLOGICAL OBSERVATIONS
Productivity
Depth
Chlorophyll
Bacteria
in situ
Incubator
B-
U- P-
(m)
( "
^mg
;/mJ)
( no /ml )
(mg C/m3/day)
(m
g c/m3/hr)
0
.
157
_
3-7
0
.0H8
-
-
25
367
-
-
0
.072
-
-
50
.
217
-
-
0
.20
-
-
100
231
-
-
0
.030
-
-
Zooplankton Volume: 37 ml/lOOO m3 total, 37 ml/lOOO m3 small.
Incident Radiation
Daily Max: I.59 cal/cm /min. Daily Total: 395 cal/cm . Day Length: 10. 60 hrs,
12
Station 2 (Cont.)
SUBMARINE DAYLIGHT (1*80 m^)
Depth
Corr. Sub.
(m)
Read.
(pa)
k/m
7
6i*o
_
12
22
530
.0376
390
.0307
37
228
.0358
72
1+8.3
.01*14-3
122
6.6
.01*08
VoTm
96.3
97-0
96.5
95.7
96.O
13
Station 3
M/V Stranger; SCOPE; November 12, 1956; 2104 GCT; 19°17.0'N, 106°32.0'W; l600 fm;
wire angle, 10°; temp., 82.0°F dry, 77.1°F wet; weather, 02; clouds, 8, amt. 3;
sea, 3j swell, confused.
OBSERVED
Depth
Temp.
s
Op
POI4.-P
N02-N
PH
Alk
(m)
C°c)
Ct)
(ml/1)
(ugm at/l)
(ligm at/l)
(millival/l)
0
28.56
34.57
4.47
O.63
_
8.21
—
23
28.20
34.1+7
4.47
0.48
-
8.22
_
33
27.33
34.40
5.6l
0.52
-
8.24
_
47
22. 1+5
34.48
3.84
O.96
-
8.13
-
%
20.22
34.51
2.51
1.39
-
8.02
_
74
11+.98
34.55
0.64
2.17
-
7.83
-
137
12.65
3^.79
-
2.38
-
7.76
_
188
11.85
34.81
0.21
2.36
-
7.76
_
234
/ 11-35
k 11.07
34.74
0.10
2.36
-
7-75
-
279
10.58
34.72
0.07
2.54
-
7.72
-
375
9.31
34.61
0.07
2.62
-
7.72
_
V71
7.88
34.54
0.06
2.74
-
7.74
-
721
5.64
34.52
0.07
2.90
-
7.71
-
BIOLOGICAL OBSERVATIONS
Depth
C»)
0
25
50
100
Chlorophyll
"a"
(mg/m3)
.204
.246
.812
.135
Zooplankton Volume
Incident Radiation
Bacteria
( no /ml )
Productivity
in situ
(mg c/nP/day)
2.1
Incubator
(mg C/m3/hr.)
O.78
0.66
0.22
0.024
32 ml/1000 m3 total, 32 ml/1000 m3 small,
B- U-
Daily Max: I.38 cal/cm /min. Daily Total: 451 cal/cm . Day Length: 10. 85 hrs,
14
Station 3 (Cont.)
SUBMARINE DAYLIGHT (h&0 mu)
Corr. Sub.
Depth
Read.
(»)
(ua)
k/m
%T/m
12
509
22
262
.066k
93.6
32
106
.0905
91.3
1+2
58.8
.0589
9^-3
52
19.1
.1124
89A
62
l.h
.O9I+9
90.9
15 -
Station *+
m/v
Stranger:
SCOPE;
; November
13, 1956; 2017 l
3CT;
17'
'27
.O'N
, 102°53
,0'Wj
;
5210 fm; wire
angle,
6°; temp.,
81+.5°F dry, 77
.0°F
wei
weather, 02;
i
:louds, 8, amt., 2; :
;ea, 1; swell, 3^0°, 3 ft,
10
sec,
OBSERVED
Depth
Temp.
S
,°2,
POi, -P
N02-N
PH
Alk
(m)
(°c)
['/..)
(ml/1)
(Hgm at/1)
(
\igm
at
A)
(millival/l)
0
29.38
3k. 3k
^•33
O.58
-
8.19
-
2k
29.02
3I+.29
k.k2
0.1+5
-
8.19
-
1+2
25.50
3I+.1+0
1+.17
0.64
-
8.17
-
52
21.78
3^. 3^
3.56
0.97
-
8.12
-
56
20. Ik
3I+.V7
2.08
1.37
-
8.00
-
80
15.27
3I+.67
0.09
2.56
-
7.78
-
121
13.17
3^.86
0.05
2.1+6
-
7.79
-
169
12.37
3I+.87
0.08
2.60
-
7.79
-
2*+l
/•II.36
V12.ll+
3^.80
0.10
2.66
-
7-lk
-
286
11.00
3^.77
0.08
2.81+
-
7-75
-
386
9.68
3^.70
0.10
2.79
-
7.7^
-
l+80
8.05
34.61
0.10
3.12
-
7-7^
-
731 N
6.01
3^.57
0.08
3.29
-
7.68
-
706^
BIOLOGICAL OBSERVATIONS
Depth
(m)
0
25
50
100
Chlorophyll
"a"
(mg/m )
.118
.130
.588
.582
Bacteria
(no /ml)
Productivity
in situ
(mg C/nP/day)
U.5
Incubator
(mg c/m3/hr._
0.13
0.17
0.23
0.11
Zooplankton Volume: 76 ml/1000 m3 total, 5^ ml/1000 m3 small.
Incident Radiation ? , 2
Daily Maximum: 1.15 cal/cnT/min. Daily Total: 1+52 cal/cm .
B-
U- P-
Day Length: 10.95 hrs.
16
Station 5
M/V Stranger; SCOPE; November ik, 1956; l8ol+ GCT; l6°l5.5'N, 100°28.0'W;
21+00 fm; wire angle, 9°; temp., 83.0°F dry, 77.0°F wet; weather, 02;
clouds, 6 and 8, amt., 2; sea, 2; swell, confused.
OBSERVED
'epth
Temp.
S
02
POl^-P
N02-N
pH
Alk
(m)
(°c)
%)
(ml/1)
(ugm at/l)
(ugm at/l)
(millival/l)
0
29.22
33.61
J+.32
0.1+6
_
8.22
2.3I+
8
29.22
33.60
k.3k
0.1+0
-
8.25
2.32
15
29.18
33.61
k.k2
O.36
-
8.25
2.3I+
2k
29.16
33-68
Ml
0.37
-
8.23
2.3I+
^3
26.80
3I+.2I+
4.27
0.52
-
8.20
2.36
52
22.19
3^- ^5
1.70
1.68
-
7-97
2.36
66
18.56
3^.58
0.31
2.02
-
7.81+
2.37
ito
12.81
3^.87
0.19
2.51+
-
7.82
2.37
190
• 12.12
3^.83
0.07
2.51+
-
7-77
2.38
285
IO.89
3^.73
0.14
2.68
-
7.72
2.37
381+
9-35
3^.67
0.13
2.92
-
7.72
2.39
1+80
7.96
3I+.62
0.1.4
3.22
-
7.72
2.1+0
728
5.91
3^.55
0.11
3.22
-
7.72
2.1+0
Depth
Chlorophyll
it 11
(m)
3
(mg/nr5)
0
0.213
5
-
10
-
15
-
25
0.162
1+0
-
50
1.02
75
-
100
0.337
BIOLOGICAL OBSERVATIONS
Productivity
Bacteria
( no/ml )
in situ
(mg C/m3/day)
Incubator
(mg c/m3/hr.)
0.15
0.16
0.13
0.079
0.056
0.10
0.033
B-
U-
Zooplankton Volume: 87 ms/lOOO m^ total, 85 ml/lOOO m small.
Incident Radiation
Daily Maximum: 1.55 cal/cm /min. Daily Total: 377 cal/cm . Day Length: 11.00 hrs.
17 -
Station 6
M/V Stranger; SCOPE; November 15, 1956; 2138, 2155 GCT; l4°17.0'N, 96°34.0'W;
1900 fm; wire angle, 30°, 35°; temp., missing; weather, 02; clouds, missing;
sea, 2; swell, missing.
OBSERVED
Depth
Temp.
S
°2
(ml7l)
P°4-p .
N02-N
pH
Alk
(m)
(°c)
%)
(u gm at/l)
(n gm at/l)
(millival/l)
0
27.67
33A2
4.55
0.65
_
8.25
19
26.68
33.78
4.36
0.65
-
8.23
_
27
25.88
33-95
3.84
0.78
-
8.19
_
30
24.36
34.04
3-^0
1.12
-
8.15
_
59
21.78
34.16
3.64
1.36
-
8.12
-
73
21.02
34.27
3-56
1.50
_
8.09
82
20.86
34.25
3.46
1.46
-
8.08
_
108
19.05
34.48
2.29
1.77
_
8.00
_
i4i
13.71
34.84
0.31
2.24
-
7.95
_
207
12.5^
34.82
0.24
2.35
-
7.87
_
276
11.53
34.79
0.08
2.24
_
7.80
_
345
10.50
34.76
0.09
2.67
-
7.77
.
536
7-75
3:'.6o
0.07
2.99
-
7.77
-
Depth Chlorophyll Bacteria
0
10
25
50
75
100
150
(mg/m )
• 577
.920
.613
.840
( no/ml )
33
8
11
49
35
38
49
BIOLOGICAL OBSERVATIONS
Productivity
in situ Incubator
Xmg C/m3/day) (mg C/m3/hr.)
0.57
B-
U-
Zooplankton Volume: 325 ml/1000 m3 total, 3l4 ml/lOOO m3 small.
Incident Radiation p
Daily Maximum: 1.55 cal/cm /min. Daily Total: 449 cal/cm . Day Length:11.25 hrs.
- 18
Station 7
m/v
Stranger;
SCOPE;
November l6j
, 1956; 19^2
GCT;
12'
'1+1,
.0'
N,
9V 15.0
•W;
2200 fm;
wire
i angle, 5
; temp . ,
81+.0°F dry,
78.8°F wet;
weather.
, 02;,
:louds, 8,
amt
., 3;
sea,
2; swell
, 3^°, k
ft. 7 sec.
OBSERVED
lepth
L
Temp.
s
02
P04-P
NO'
>-J
[
pH
Alk
(m)
(°c)
' /•»)
(ml/D
(ugm at/l)
(ugm~
'at
■/U
(mi!
Llival/l)
0
27.93
33.62
4.6l
1.00
_
8.19
_
12
27.66
^.65
0.88
-
8.21
-
15
2^.72
33.9^
3.29
1.19
-
8.11
-
IT
20.1+8
3A.56
1.77
1.83
-
7-99
-
19
19.16
3^.57
1.99
I.83
-
8.00
-
29
16.92
3^.65
0.31
2A9
-
7.83
-
58
13.78
3^.83
0.0*+
2.5I+
-
7.82
-
lll+
12.52
3^.82
0.07
2.32
-
7.81+
-
193
11.55
3^.76
0.10
2.60
-
7.80
-
287
10. oi«-
3^.70
0.10
2.80
-
7.78
-
386
8.1+1+
3 k. 61
0.14
2.99
-
7-75
-
ATT
7.k6
3^.59
0.12
3.17
-
7.76
-
729
5.58
3^.5^
0.11
3.01
-
7.73
-
BIOLOGICAL OBSERVATIONS
Productivity
epth
Chlorophyll
Bacteria
"a"
in situ
(m)
(mg/m )
(no /ml)
(mg C/nP/day)
0
.380
51+6
_
10
-
59
-
25
3.76
12
-
50
1.12
12
-
75
-
19
-
100
-
56
-
Incubator
(mg c/nrW.)
1.1
B-
U-
Zooplankton volume: 192 ml/1000 mJ total, 192 ml/lOOO m small.
Incident Radiation 2
Daily Max: 1.30 cal/cnr/min. Daily Total: 1+1+7 cal/cm . Day Length: 11.20 hrs.
- 19
Station 8
M/V Stranger; SCOPE; November 17, 1956; 1935 GCT; 11°13.0'N, 90°55.0'W; 19^0 fm;
wire angle, 20°; temp., 8l.5°F dry, 76.2°F wet; weather, 01; clouds, 8, amt., 6;
sea, 3; swell, 110°, 5 ft, 6 sec.
OBSERVED
Depth
Temp.
S
°2
POirP
N02
-N
PH
Alk.
(m)
(°0)
I /••)
(ml/1)
1
[\i gm at/l)
(U gm
at/l)
(.
nillival/ljl
0
27.58
33.27
1J-.71
0.70
0.1
8.21+
2.31
8
27.5^
33-21+
1+.75
0.71
0.1
8.21
2.31
15
27. in
33.26
k.8k
0.66
0.1
8.22
2.29
19
27.26
33.28
1+.51
0.79
0.1
8.21
2.29
28
22.06
3I+.I+2
2. 91+
1.22
0.8
8.03
2.39
51
17.02
31+.87
1.03
2.1I+
0.2
7.89
2.36
87
13.67
3^.92
0.30
2.23
t
7.86
2.37
120
13.014.
31+. 92
0.35
2.17
t
7.87
2.37
182
12.26
3I+.87
0.35
2.21+
0.0
7.86
2.36
277
10.56
-
0.08
2.1+2
0.1
7-77
2.38
37^
8.1*2
3^.63
0.08
2.57
1.3
7.75
2.1+3
1+70
T.kh
3^.61
0.10
2.89
1.0
7-7*+
2.1+0
718
5.60
3^.57
0.11+
2.90
0.1
7.78
2.1+0
BI0L0GICAL
OBSERVATIONS
P
roductivity
Depth
Chlorophyll Bacteria
B- U-
P-
"a"
in
situ.
Incubator
(»)
(mg/m )
( no /ml
) Tmg
C/m-
/day) (mg
C/m^ hr.)
0
.582
_
_
_
_ _
_
25
.980
-
-
-
-
-
50
.762
-
-
-
"
-
100
.118
-
-
-
-
-
Zooplankton Volume: 125 ml/lOOO nr5 total, 125 ml/1000 m3 small.
Incident Radiation
Daily Max: 1.59 cal/cm /min. Daily Total: 1+1+8 cal/cm . Day Length: 11.1+3 hrs.
20 -
Station 9
November l8, 1956
08°56' N 88°30' W
NO CAST
BIOLOGICAL OBSERVATIONS
Productivity
Depth
Chloro
"a"
(m)
(mg/m
0
.336
9
.426
15
• 330
50
.300
100
.112
in situ Incubator
(no/ml) (mg C/m3/day) (mg C/m3 hr.
13.0 2.0
B-
P-
Zooplankton Volume: 95 ml/1000 nr total, 95 ml/lOOO nr small.
Incident Radiation _ ?
Daily Maximum: 1.42 cal/cm /min. Daily Total: 511 cal/cm . Day Length: 11.51 hrs.
21
Station 9A
M/V Stranger; SCOPE; November 19, 1956; 0159 GCT; 08°56.0'N, 88°29.5'W; 2100 fm;
wire angle, 15°; temp., T8.0°F dry, 73-2°F wet; weather, 02;clouds, 3, amt., 3;
sea, 4; swell, confused.
OBSERVED
Depth
Temp.
s
02
po4"p .
NO2-N
PH
Alk
(m)
(°c)
(7oo)
(ml/D
(ngm at/1)
(u.gm at/l)
(millival/l)
0
25.72
33-37
4.27
1.19
0.2
8.19
2.27
5
r25.99
^25.72
33.39
4.26
1.05
0.1
8.20
2.30
9
24.60
33.60
4.02
1.33
0.2
8.15
2.31
12
22.18
34.07
3.26
1.70
0.2
8.08
2.32
22
18.28
34.65
I.78
2.11
0.5
7-99
2.36
27
17.04
34.69
1.38
2.19
0.5
7.88
2.35
63
13.97
34.92
0.70
2.16
t
7.91
2.36
111
13.00
34.90
0.48
2.24
t
7.86
2.37
202
11.94
34.85
0.54
2.39
0.1
7.83
2.37
308
IO.78
34.75
0.21
2.53
t
7-79
2.37
>H3
9.49
34.70
0.10
2.69
t
7.76
2.38
5i4
7-97
34.63
0.10
2.97
0.2
7.75
2.38
767
5.70
34.59
0.30
2.96
0.1
7-73
2.40
1035
4.47
34.61
0.83
3.08
0.0
7.82
2.4l
1559
3.08
34.61
-
-
0.0
7.85
2.44
BIOLOGICAL OBSERVATIONS
Productivity-
Depth
(m)
0
2
10
27
40
Chlorophyll
"a"
(mg/nr1)
Bacteria
( no/ml )
in situ
(mg C/mJ/day)
6.1+
Incubator
(mg C/nr hr.)
0.77
0.70
0.50
0.30
0.33
U-
P-
Zooplankton Volume: 95 ml/1000 m3 total, 95 ml/lOOO m3 small.
Incident Radiation
Daily Max: 140 cal/cm /min. Daily Total: 535 cal/cm2. Day Length: 11. 58 hrs.
22
Station 9C
M/V Stranger; SCOPE; November 20, 1956; 1942 GOT; 09°15.5'N, 89°l8.0'W; l820 fm;
wire angle, 0°; temp., 79.5°F dry, 76.28F wet; weather, 02; clouds, 4-5; amt., 6;
sea, 2; swell, 06o°, 3 ft, 4 sec.
OBSERVED
Depth
Temp.
S
°3
WD
P°4-p .
N02-N
PH
Alk
(*)
(°c)
("/••)
(ugm at/1)
(ugm at/l)
(millival/l)
0
25.26
33.67
4.04
0.92
0.2
8.17
2.33
6
24.81
33.64
4.11
1.04
0.1
8.18
2.32
8
24.65
33.71
4.05
O.96
0.1
8.16
2.31
11
24.41
33-68
3.91
1.03
0.1
8.15
2.32
13
22.66
33-86
3-39
1.20
0.2
8.12
2.33
15
20.92
34.24
2.82
1.40
0.2
8.06
2.34
17
17.74
34.71
1.53
1.79
0.3
7.97
2.36
19
-
34.77
1.25
I.78
0.3
7-93
2.37
21
16.42
34.78
1.26
I.89
0.1
7-93
2.37
23
15.98
34.79
1.01
I.87
0.4
7.92
2.37
25
15.22
34.78
0.87
2.04
0.5
7.92
2.37
28
14.48
34.78
0.88
2.17
0.5
7.92
2.36
30
14.27
34.83
0.79
2.04
0.5
7.91
2.37
k9
13.61
34.79
0.66
2.15
0.4
7.90
2.37
73
12.70
34.79
1.13
2.20
0.1
7-95
2.37
97
12.17
34.78
1.16
2.27
t
7-95
2.37
l4l
II.98
34.86
0.71
2.42
0.0
7.92
2.38
BIOLOGICAL OBSERVATIONS
Productivity
Depth
(m)
0
5
8
10
12
14
16
18
20
22
27
30
50
100
Chlorophyll
"a" 3
(mg/m )
.191
.231
.381
.228
.446
.302
.319
.188
Bacteria
( no /ml )
in situ
(mg C/m^/day)
13
11
13
15
14
10
4.5
4.8
3.2
1.7
O.29
Incubator
(mg c/nrhr.)
0.70
O.98
0.91
0.56
0.16
23
Station 9C (Cont.)
Water Column Productivity: O.332 mgC/m /day
Zooplankton Volume: 250 ml/lOOO m3 total, 250 ml/lOOO m3 small
Incidental Radiation
o
Daily Maximum: I.83 cal/cm /min.
Daily Total: 399 cal/cm2.
Day Length: 11.1+9 hrs.
SUBMARINE DAYLIGHT (1*25 mu)
Corr. Sub
Depth
Read.
(m)
(ua)
k/m
°/»T/]
2
382.0
_
_
12
188.0
.0798
93-2
22
72.1+
.0951*-
90.9
32
25.6
.103
90.1
1+1
6.3
.155
85.6
2U-
Station 9D
M/V Stranger; SCOPE; November 21, 1956; 2005 GCT; 09°31+.0'N, 89°13.5'W; l800 fm;
wire angle, 0°; temp., 80.0°F dry, 76.8°F wet; weather, 01; clouds, 4 and 8, amt.4;
sea, 1; swell, 110°, 2 ft, 9 sec.
OBSERVED
Depth
(m)
0
10
12
14
16
18
20
22
24
27
30
1*9
73
77
145
Temp. S 02 PO^-P
(°C) (%.) (ml/1) (ngm at/1)
,25.51 33-61+ I1.11 1.1U
l25.29
2I4-.9B 33.63 ^.11 1-23
2U.90 33.64 4.09 l.ll
24.83 33.84 U.03 1.46
24.74 33.67 4.01 1.28
24.53 33-68 3.98 1.18
23.82 33.80 3.60 1.23
22.63 33.96 3.29 1.36
20.65 34.30 2.54 1.70
18.86 34.51 1.89 1.85
18.11 34.69 1.67 1.83
17.18 34.72 1.13 2.04
16.82 34.73 1.18 2.04
14.36 34.86 O.98 2.20
13.31 34.87 0.74 2.53
12.86 34.87 0.48 2.60
12.30 34.87 0.44 2.21
N02-N
(ugn at/l)
0.1
0.2
0.3
0.3
0.3
tr.
0.0
tr.
0.0
PH
8.12
8.16
8.16
8.16
8.16
8.14
8.15
8.11
8.05
7-99
7-99
7-93
7-93
7.94
7-9^
7.88
7.85
Alk
(millival/l)
BIOLOGICAL OBSERVATIONS
Productivity
Depth
(m)
0
5
8
10
12
14
16
18
20
22
27
30
50
100
200
Chlorophyll
"a"
(mg/m )
.308
.310
.3^0
.342
• 335
.458
.280
.092
Bacteria
B-
U-
In situ Incubator
(no /ml) Jmg C/nr/day) (mg C/nrhr.)
3-2
8.1
4.6
3-7
2.5
7-3
6.2
10
9.4
3-0
0.38
0.52
+
+
0
0
0
0
0.79
-
-
-
0
0
0
-
-
-
-
-
+
0
0
0.85
0
0
0
-
-
-
-
~
+
+
0
-
0
0
0
0.31
0
0
0
-
0
0
0
-
0
0
0
25
Station 9D
( cont . )
o
Water Column Productivity = 0.1+02gmC/m / day.
Zooplankton Volume: 135 ml/lOOO nT total, 135 ml/lOOO nr small.
Incident Radiation
Daily Maximum: 1.66 cal/cm /min.
Daily Total: ^55 Cal/cm2.
Day Length: 11. ^ hrs.
SUBMARINE DAYLIGHT (h8o mn)
Corr. Sub.
Depth Read.
(ua) k/m VoT/m
2 202 -
167 .0380 96.3
135 .01+25 95.8
21 69.1 .07I4.1+ 92.8
28 1+0.2 .0773 92.6
29.8 .0332 96.7
15.6 .0719 93.0
26
Station 9F
M/V Stranger;
wire angle, 3
sea,
SCOPE;
temp.,
1; swell, l80°, 3 ft,
November 23, 1956; 1332 GCT; 09°4l.0'N, 89°44.5'W; 1700 fm;
79.0°F dry, 77.2°F wet; weather, 02; clouds, 6, amt., 5;
sec.
Depth
(m)
0
4
8
10
12
13
16
18
20
22
24
26
30
50
75
99
11+8
Temp . £
CO (%
25.44
25.36
25.22
25.14
2U-.99
24,87
24.56
24.26
23 .1*9
22. 45
18.77
17-75
17. 0^4-
15.20
13.58
13.01
12.76
12.16
33.64
33.63
33.63
33-68
33.68
33-68
33-71
33-75
33.81
34.00
34.31
34.60
34.79
34.87
34.92
34.92
34.87
02
(ml/1)
4.20
4.02
4.16
4.02
3-99
3.94
3.82
3.74
3.56
3.17
2.41
1.58
0.94
0.37
0.49
0.51
O.56
OBSERVED
PO^-P
(ngm at/l)
1.06
1.14
1.16
1.04
1.04
1.14
1.05
1.12
1.20
1.36
1.57
1.78
1.96
2.17
2.12
2.18
2.15
N02-N
(ngm at/l)
PH
Alk
(millival/l)
BIOLOGICAL OBSERVATIONS
Productivity
Depth
(m)
0
2
5
8
10
12
14
18
20
22
30
50
100
Chlorophyll
"a" 3
(mg/m )
.130
.276
• 3^3
.387
.284
.011
Bacteria
(no /ml)
in situ
(mg C/mJ/day
7.0
7-3
7-7
5.6
5.7
4.4
12.0
14.0
11.0
5.0
1.4
U-
P-
Incubator
(mg C/m3 hr . )
0.59
0.70
0.44
0.58
0.46
O.74
0.20
Water Column Productivity = 0.320gmC/m day
Incident Radiation ? ?
Daily Max: 1.43 cal/cm /min. Daily Total: 482 cal/cm . Day Length: 11.45 hrs,
27 -
Station 9F (Cont.)
SUBMARINE DAYLIGHT (H8o njl)
Corr. Sub.
Depth
Read.
(m)
(na)
7
967
12
708
22
335
31
159
in
82.2
50
40.8
6o
12.6
66
4.3
71
3.2
k/m °/oT/n
.0623 93-9
.O7U8 92.8
.0828 92.O
.0659 93-6
.0778 92.5
.117 88.9
.179 83.6
.0590 9^.3
- 28
Station 10
M/V Stranger; SCOPE; November 2k, 1956; 1912 GCT; 08°1+2.0'N, 86°01.0'W; l650 fm;
wire angle, 3°; temp., 80.0°F dry, 75.6°F wet; weather, 02; clouds, 6, amt., 7;
sea, 3; swell, 120°, 3 ft. 6 sec.
OBSERVED
epth
Temp.
s
Oo
(ml/D
PCVP .
N02-N
pH Alk
(m)
(°c)
(%«)
(ugm at/l)
(ugm at/l)
(milllval/l
0
26.86
32.65
I4-.56
0.65
tr.
8.2^
20
26.79
32.68
1+.55
0.52
tr.
8. 21+
30
25.9^
33.11
1+.20
O.67
0.1
8.23
37
23.00
33.93
2.7I+
1.37
0.3
8.09
k2
19.5^
31+.1+9
1.91
1.80
0.5
7.98
51
16.92
3^.7^
1.30
1.97
1.0
7-9^
96
13. I**
31+.87
O.7I+
2.18
tr.
7.89
lJ+2
12.5^
3^.83
0.62
2.25
0.1
7.89
196
11.9^
31+.83
0.55
2.33
0.0
7.86
290
10.72
3^.7^
0.30
2.1+0
tr.
7.78
388
9.06
3^.67
0.08
2.79
0.7
7.76
k8k
7.85
3^.65
0.10
2.81+
0.0
7-75
736
5.78
3I+.60
-
2.96
tr.
7.86
epth
Chlorophyll
(m)
"a"
(mg/m )
0
.1+20
10
.1+11+
25
• 55^
50
• 759
100
.227
BIOLOGICAL OBSERVATIONS
Productivity
Bacteria
in situ
(no/ml) (mg C/m-^/day)
28.0
Incubator
(mg C/m3/hr.)
1.0
1.2
O.89
1.0
0.32
Zooplankton Volume; l66 m1/l000m3 total, 166 ml/l000m3 small.
Incident Radiation p „
Daily Max: O.377 cal/cm /min. Daily Total: 121 cal/cm
B-
U-
Day Length: 11.20 hrs.
SUBMARINE
DAYLIGHT
(l+80 mu)
Depth
Corr. Sub.
, Read
(m)
(ua)
k/m
°/0T/m
2
221+
7
167
.0587
91+.3
12
107.5
.0871
91.6
22
61+. 9
.0509
95.0
32
31.6
. .0719
93-0
1+2
12.01+
.0968
90.8
52
3.20
.132
87.6
29
Station 11
M/V Stranger; SCOPE; November 25, 1956; 1932 GCT; 07°37.0'N, 82°25.5'W; 600 fa;
wire
angle,
15°; temp. ,
, 74.8°F dry,
73.8°F wet;
weather, 2C
; clouds ,
, 9, amt., 7;
sea,
2; swell, 120°, 2
ft., 3 sec.
OBSERVED
Depth
Temp.
S
02
P04-P
N02-N
pH
Alk
(m)
CO
(%o)
(ml/D
(ngm at/l)
(jigm at/l)
(millival/l)
0
26.72
29.1+0
-
0.44
0.0
8.27
-
15
26.76
29.70
-
0.4l
0.0
8.25
-
30
25.82
31.82
-
O.98
0.0
8.22
-
44
23. Ok
33-1*8
-
1.00
1.5
8.12
-
53
18.70
34.51
-
I.69
0.2
7.96
-
59
17.03
3I+.78
-
1.84
0.1
7.92
-
91
15.22
34.88
-
2.21
tr.
7.88
-
127
1A.3A
34.93
-
2.11
tr.
7.84
-
205
12.68
3I+.89
-
2.32
0.0
7.80
-
300
11.12
34.82
-
2.58
0.0
7.76
-
iK)6
9.32
3I+.69
-
2.82
0.0
7.71
-
507
7. 97
34.65
-
3.06
0.0
7-71
-
769
5.83
34.59
-
3.09
0.0
7.71
-
Depth
(m)
0
10
25
50
100
BIOLOGICAL OBSERVATIONS
Productivity
Chlorophyll
"a"
(mg/m )
• 517
.526
.734
1.20
.186
Bacteria
( no /ml )
in situ
(mg cjnf/dsy)
Incubator
(mg C/nrVhr.)
0.59
0.51
1.3
1.0
0.15
Zooplankton Volume: 104 ml/lOOOnr total, 104 ml/lOOOm^ small.
Incident Radiation p ?
Daily Maximum: 0.977 cal/cm /min. Daily Total: 83.2 cal/cm .
U-
P-
Day Length:
11.43 hrs,
Depth
(m)
2
7
SUBMARINE DAYLIGBT (48o mn)
k/m °/.T/]
.238 78.8
Corr. Sub. Read
(na)
m
10
33
- 30
Station l6
M/V Stranger; SCOPE; December 1, 1956; l835 OCT; 05°59.0'N, 79°48.8'W; 1700 fm;
wire angle, 13°; temp., 82.3°F dry, 75.8°F wet; weather, 02; clouds, 4, amt. 6;
6ea, 1; swell, confused.
OBSERVED
Depth
(m)
0
6
18
29
39
43
52
94
190
283
379
1+72
720
Temp.
CO
26.68
26.50
26.62
25.94
24.32
23.03
19 A3
14.34
13.16
11.94
9.37
8.42
6.09
(V..)
28.30
28.38
30.61
33.11
33.61
33-93
34.66
34.96
34.97
34.88
34.72
34.69
34.60
°2
(ml7D
4.52
4.57
40
35
10
42
13
0.85
0.59
0.33
0.24
O.17
0.57
P01+-P
(ugm at/l)
0.30
0.20
0.18
0.31
0.50
0.73
NO
16
1.60
1.58
1.92
2.02
1.68
2.18
U-N
1 at/l)
pH
Alk
(milllval/l)
tr.
8.24
_
0.0
8.24
-
tr.
8.24
-
0.0
8.23
-
0.1
8.16
-
0.7
8.12
-
0.4
8.01
-
0.0
7.92
-
0.0
7.88
-
0.0
7.79
-
tr.
7.76
-
0.0
7.76
-
0.0
7.78
-
BIOLOGICAL OBSERVATIONS
Depth
(m)
0
10
25
50
75
100
Chlorophyll
"a"
(mg/nr1)
.329
.272
.364
.491
.101
Bacteria
(no /ml)
75
3
8
1
1
Productivity
in situ
Incubator
0.14
B-
Zooplankton Volume: 95 ml/lOOO m3 total, 95 ml/lOOOm3 small.
Incident Radiation ? 2
Daily Max: 2.13 cal/cm /min. Daily Total: 437 cal/cm . Day Length:
U-
P-
(mg c/m:
Vday)
(mg
C/m3/hr.)
13.0
O.38
0
0
0
_
O.98
0
+
-
_
0.30
-
-
-
-
0.23
0.26
0
+
0
0
0
11.60 hrs.
SUBMARINE
DAYLIGHT
(480 mu)
Depth
Corr. Sub.
Read.
(m)
(ua)
k/m
7.T/>
2
817
_
_
7
576
.0699
93-3
12
440
.0538
94.8
22
265
.0507
95.0
31
175
.0461
95.5
4l
74.9
.0848
91.9
49
44.8
.0642
93.8
54
33.0
.0611
94.1
59
24.0
.0636
93.8
67
12.6
- 31 -
.0795
92.3
Station 17
M/V Stranger; SCOPE; December 2, 1956; 19^9 GCT; 04°09.0'N, 83°34.0'W; 1700 fm;
wire angle, 0°; temp., missing; weather, 02; clouds, 8, amt., 5; sea, 1;
swell, slight.
OBSERVED
Depth
Temp.
S
02
POI4.-P
N02-N
pH
Alk
(m)
(°c)
(V..)
(ml/1)
(ligm at/l)
(|igm at/l)
(millival/l)
0
27.02
32.96
4.43
0.46
tr.
8.19
2.24
9
26.1*2
32.95
4.50
0.40
tr.
8.21
2.24
18
26.38
33.01
4.48
0.43
tr.
8.22
2.24
27
25.54
33.36
4.18
0.59
0.1
8.20
2.25
30
21*. 1+5
33-61*
3-9^
0.71
0.2
8.15
2.28
37
20.64
3^-31
2.78
1.05
0.5
8.07
2.31
49
18.50
3l*.70
2.15
1.26
0.7
8.00
2.34
97
14.76
31*. 97
1.46
1.82
tr.
7.93
2.35
197
13.22
31*. 96
O.63
1.66
0.0
7.84
2.35
292
11.71*
34.88
0.33
1.87
tr.
7.80
2.35
394
9.3^
34.70
0.14
2.13
tr.
7-7^
2.36
490
8.05
3^.67
0.22
2.27
tr.
7.75
2.37
743
5.86
34.61
O.69
2.32
tr.
7.82
2.38
Depth Chlorophyll Bacteria
(m) (mg/m0)
0 .196
5
10 .215
25 .261
50 .633
75
100 .105
BIOLOGICAL OBSERVATIONS
Productivity
in situ
Incubator
( no/ml )
23
6
5
2
23
5
2
(mg C/m3/day) (mg C/m3/hr.)
6.1 0.35
0.37
0.47
O.36
B-
0.070
Zooplankton Volume: 139 ml/l000m3 total, 139 ml/lOOOm3 small.
Incident Radiation _
Daily Max: 1.80 cal/cm /min. Daily Total: 433 cal/cm . Day Length
SUBMARINE DAYLIGHT (480 mu)
U-
P-
0
0 0
■++
0 0
-H-
+ 0
+*
+ 0
-H-
+ 0
+
0 0
+
0 0
igth
: 11.86 hrs
Depth
Corr. Sub. Read.
(m)
(na)
k/m
%T/,
2
921
-
-
7
727
.0473
95.4
12
607
.0360
96.4
22
4l6
.0377
96.2
32
288
.0367
96.4
4l
135
.0841
91.9
51
63.8
.0749
92.8
61
27.7
.0834
92.0
69.5
16.7
.0632
93-9
77
13.2
• 0335
96.7
32
Station l8
M/V Stranger; SCOPE; December 3, 1956; 20l*9 GCT; 05°28.5'N, 86°57.0'W; 700 fmj
wire angle, 5°; temp., 79.8°F dry, 76.0°F wet; weather, 02; clouds 8, amt . 6;
sea, 2; swell, 210°, 3 ft . 5 sec .
OBSERVED
Depth
Temp.
S
°2
(ml7D
PO^-P
N02-N
pH
Alk
(m)
(°c)
(*/-)
(*igm at/1)
(ugm at/l)
(millival/l)
0
26.1+2
33.17
»*.1*5
0.30
tr.
8.22
2.27
15
26.10
33-15
14-.51
0.28
0.0
8.25
2.27
30
25.95
33.28
^.56
0.30
0.0
8.26
2.27
in
21):. 05
33-73
1+.03
-
0.2
8.20
2.29
1*7
21.70
3^.36
3.19
0.88
0.5
8.10
2.33
56
18.05
3^-9^
2.13
1.18
0.1+
8.00
2.36
91
16.76
35.03
1.93
1.28
tr.
8.00
2.36
121*
1I+.96
3^.96
1.18
1.57
tr.
7.96
2.36
199
13.16
3^-97
0.52
1.72
0.0
7.86
2.36
292
12. 1*1
3^.90
0.1*3
1.80
0.0
7.81*
2.36
391
11.06
3^-79
0.21
2.00
0.0
7.78
2.36
1*85
8.97
3^.70
0.12
2.28
0.0
7.78
2.37
736
6.07
3^-59
0.1*4
2.1*9
0.0
7.80
2.39
BIOLOGICAL OBSERVATIONS
Productivity
Depth
Chlorophyll Bacteria
"a"
in situ
Incubator
B-
U- P-
Cm)
0
(mg/m )
(
no /ml)
(mg c/nr/day)
(mg C/m3/hr.)
0
.169
111*
3-8
0.21+
+
0 0
10
.226
23
-
0.30
-
-
25
.329
19
-
O.3I*
-
-
50
.1*25
6
-
0.1*6
-
-
75
-
161
-
-
-
-
100
.230
8
-
0.11
-
-
3 o
Zooplankton Volume: 111* ml/i000m total, 111* ml/lOOOnr small.
Incident Radiation ? 2
Daily Max: I.9I* cal/cm /min. Daily Total: 279 cal/cm . Day Length, 11.1*5 hrs .
SUBMARINE DAYLIGHT (1*80 m*x)
epth
Corr. Sub. Read
(m)
(*ia)
k/m
%T/m
2
877
-
-
7
763
.0278
97-3
12
690
.0201
98.0
22
1*18
.0501
95-1
30
268
.0555
91*. 6
1*1
123
.0708
93-2
51
61
.0701
93-2
61
37-2
.0l*9l*
95.2
70
15.9
.091*1*
91.0
33
Station 19
v Granger; SCOPE; December k, 1956; 1956 GOT; 06o46.0'N, 89°52.0'W; 1900 fm;
wire sr.gle, 10°; temp., 82.5°F dry, 77.1°F wet; weather, 02; clouds, 8, amt., 3;
sea, 1; swell, 270°, 3 ft, 6 sec.
OBSERVED
rr- .
Temp.
S
Op
(ml7D
PO^-P
NO2-N
pH
Alk
(■)
(°c)
("/••)
(ugm at/l)
(ugm at/l)
(millival/l)
0
27.06
32.51
4.39
0.30
tr.
8.22
2.22
7
26.74
32.51
4.37
0.3I*
0.0
8.23
2.21
18
26.50
32.95
4.33
O.36
tr.
8.22
2.23
24
23.92
33.88
3.06
0.90
0.2
8.12
2.30
29
21.06
34.53
2.53
1.11
0.1+
8.02
2.33
35
17.83
34.66
1.16
1.6l
1.1
7.89
2.33
47
15.1*6
34.88
i.l+O
1.1*8
0.7
7.92
2.31+
94
13.57
34.97
O.98
1.58
tr.
7.89
2.35
190
12.66
34.88
0.73
1.66
0.0
7.86
2.35
283
11.52
34.85
0.58
1.76
0.0
7.82
2.35
380
10.21*
34.74
0.62
1.86
0.0
7-79
2.35
474
-.: =
34.69
0.17
2.16
tr.
7.75
2.37
722
6.03
3^.6o
0.13
2.1+4
0.0
7.76
2.38
BIOLOGICAL OBSERVATIONS
Productivity
Dept h Chlorophyll
Bacteria
"a"
in situ
Incubator
(m) (mg/m )
( no/ml )
(mg C/nP/day)
(mg C/nrVhr.)
0 .155
53
4.5
0.19
10 .175
l*
-
0.12
25 .212
:-
-
0.23
50 .1+03
3
-
O.O76
75
10
-
_
100 .21+0
3
-
0.082
Zooplankton Volume:
97 ml/1000 nr3 total,
96 ml/1000
3
m small .
Incident Radiation
0
2
:r. . Day _e
Daily Max: I.56 cal/cm^/min.
Daily Total;
536 cal/c
SU3MARINE DAYLIGHT
(480 mu)
Depth
Corr. Sub.
Read.
(m)
(na)
k/m
7
888
_
12
723
.0411
22
526
.0318
32
262
.0696
1+0
143
.0756
50
70.6
.0705
60
36.1
.0673
68
19-5
.0766
77
13.1
.0442
86
5.4
.0984
ti-
ll.72 hrs.
VcT/m
96.0
96.9
93-3
92.7
93-2
93-5
92.6
95.7
90.6
34 -
Station 20A
M/V Stranger; SCOPE; December 5, 1956; l620 GCT; 07°50.0'N, 91°17.0'W; 1900 fm;
wire angle, 7°; temp., 79-8°F dry, 76.1°F wet; weather, 02; clouds, 8, amt., 5;
sea, 2; swell, 060°, k ft, 5 sec.
OBSERVED
Depth
Temp.
S
°2
WD
P01+-P
N02-N
PH
Alk
(m)
(°c)
(%.)
(ugm at/l)
(ugm at/l)
(millival/l)
0
25.68
33-19
I+.29
0.72
0.1
8.10
2.26
2
25-70
33-15
1+.11+
0.61+
0.1
8.11
2.25
a
25.72
33.17
1+.28
0.66
0.1
8.11
2.2b
6
25.63
33.17
1+.12
O.67
0.1
8.11
2.26
8
25.1+8
33-15
1+.20
O.67
0.1
8.11
2.26
10
25.16
33.22
1+.08
O.76
0.1
8.09
2.27
12
21+A9
33-39
3-79
0.81+
0.1
8.06
2.27
Ik
_
33-93
2.77
1.10
0.1
-
2.30
16
19.25
31+.1+0
1.8l
1.1+2
0.2
7.89
2.33
17
18.30
3^.59
1.58
1.1+8
0.2
7.88
2.3!+
19
17.1+2
3I+.69
1-33
1.52
0.2
7.87
2.3^
21
_
31+.72
1.20
1.57
0.2
-
2. 31+
23
16.19
3^-79
1.11
1.68
:•■
7.81
2.31+
27
15.70
3^-79
0.81
l.7h
0.2
7.78
2.3I+
31
_
3I+.76
0.71
1.85
0.1
-
2.35
U8
13.90
3I+.83
0.53
1.75
0.0
7-77
2.35
91+
12.90
3!+.83
0.59
1.83
0.0
7.78
2.35
BIOLOGICAL OBSERVATION
Productivity
Depth
Chlorophyll
Bacteria
B- U- I
"a"
in situ.,
Incubator
(m)
(mg/m ;
(no /ml)
Xmg C/m^/day)
(mg C
l/m3/hr . )
0
.129
97
11
O.96
-H- 0 (
1+
• 33^
37
31
-
_
c
.277
37
6.0
1.1
_
10
-
-
13
_
_
12
.382
25
8.5
_
_
11+
-
-
1+.8
-
.
15
.290
-
_
_
_
16
_
16
„
_
_
18
.1+15
-
3-7
_
_ "" .
20
-
18
.
_
24
-
20
2.1
0.32
_
25
.1+51+
-
_
_
_
30
-
6
0.38
-
_ —
50
.265
1+
0.33
0.11
..
100
.052
55
-
_ ™
Water Column Productivity: 0.270gmC/m~/day.
Incident Radiation ? P
Daily Max: 1.61+ cal/cm /min. Daily Total: 5l6 cal/cm . Day Length: 11.66 hrs.
35
Station 20B
M/V Stranger; SCOPE; December 6, 1956; 0206 GCT; 07°52.0'N, 91°19.0'W;
1900 fni; wire angle, 15°; temp., 78.1°f dry, 74.8°F wet; weather, 02;
swell, 050°, 3 ft, 5 sec.
OBSERVED
Depth
r.7emp.
S
°2
P0L.-P
TO2-N
pH
A^.k
(m)
(°c)
C/..)
(ml/1)
(ngm at/l)
(u-gm at/l)
(millival/l)
0
25.60
33.21+
4.20
0.73
0.1
8.20
2.29
6
25.62
33-24
4.22
0.68
0.1
8.20
2.28
15
23.35
33.65
3.36
0.93
0.1
8.16
2.30
18
19.31
34.51
1.68
1.34
0.2
6.05
2.34
21
17.66
34.67
l.4i
1.19
0.2
8.00
2.36
1+9
13.99
34.78
0.58
1.71
0.1
7.94
2.36
98
12.70
34.88
0.37
1.77
tr.
7.89
2.37
. 194
11.31
34.83
0.80
1.72
0.0
7.92
2.37
285
10.58
34.78
O.80
1.79
0.0
7.91
2.37
379
9.56
34.71
0.26
2.17
tr.
7.88
2.37
1+68
8.39
34.70
0.17
2.29
0.0
7.86
2.38
705
6.32
34.61
0.45
2.42
0.0
7.85
2.39
951
5.03
34.60
0.46
2.1+3
-
7.91
2.4l
1411
3.42
34.64
1.25
2.28
-
7.90
2.1+5
1915
2.46
34.69
1.98
2.19
-
7.90
2.1+3
BIOLOGICAL OBSERVATIONS
Productivity
Depth Chlorophyll Bacteria
"a" in situ Incubator
(m) (mg/nr) (no /ml) "(nig C/iiP/day) (mgC/m^/hr.)
0 .345 - 4.5
Zooplankton Volume: l46 ml/lOOOm3 total, l43 ml/l000nr small.
SUBMARINE DAYLIGHT ( 480 mu)
B-
Depth
Corr . Sub .
Read.
(m)
(na)
k/rn
7.T/1
2
981
7
682
.0727
93.0
12
576
• 0337
96.7
17
308
.125
88.2
2i+
197
.O631
93.9
III
u-
36 -
Station 21
M/V Stranger; SCOPE; December 7, 1956; 2139, 2209 GCT; 12*i7.0% ~6°50.0'W;
2200 fm; wire angle, 20°, 25°; temp,, S0.7°F dry, 75-2°F wet; weather, 02;
sea, 3.
OBSERVED
Depth
Temp.
S
02
pcv-p
N02-n
pH
Aik
(m)
(°c)
(*/..)
(ml/1)
(ugm at/l)
(ngm at/l)
(millival/l
0
25.144
33.61
1+.56
O.56
0.1
8.23
-
8
25.^0
33.61
^.55
0.62
0.1
8.23
-
31
23.82
33-93
1+.1+8
0.59
0.2
8.13
-
38
21.70
31+.23
k.2k
0.79
0.6
8.13
-
52
19.98
31+. 1+3
3.92
0.95
0.5
8.06
-
67
18.03
3^-5^
2.59
1.21
0.8
7.98
-
75
16.89
3^.65
1.86
1.52
0.8
7-95
-
129
12.61+
3I+.89
O.29
1.1+9
0.0
7.80
-
205
11.60
3^.8l
0.11
1.88
0.1
7-77
-
300
10.22
3^.7^
0.13
2.01
0.1
7.75
-
1+02
l8.6lt
3^.65
0.95
2.21
1.2
7-7^
-
1+98
7.18
3I+.61
0.11
2.29
0.6
7.76
_
736
5.Mf
3^.56
0.10
2.36
0.0
7-77
-
BIOLOGICAL OBSERVATIONS
P
roductivity
Depth
Chlorophyll Bacteria
B-
U- P-
"a"
in
situn
Incubator
(mg C/nr/hr.
(m)
(mg/m )
(no
/ml)
Tmg
C/mj/day)
■ )
0
Lost
7
_
_
-H-
0 0
10
.877
11+
-
-
-
-
25
.905
1+8
-
1.25
++
0 0
50
1.1+0
3^
-
0.95
-
-
75
-
18
-
-
-
-
100
Lost
183
-
-
-
-
Zooplankton Volume: 208 ml/l000nr total, 20l+ ml/lOOOm small,
SUBMARINE DAYLIGHT (1+80 up)
Depth
Corr. Sub.
Read.
(m)
(ua)
k/m
%T/m
2
580
_
_
7
356
.0976
90.7
12
230
.0873
91.6
21
65.2
.126
88.1
26
26.5
.180
83.5
- 37 -
Station 22
M/V Stranger; SCOPE; December 8, 1956; 1935 GCT; ll+°37.0'N, 100°09.0'W; 2000fm;
wire angle, 30°; wind, calm; temp., 85.2°F dry, 77'3°F wet; weather, 02; clouds, 1,
amt., 1; sea, 1; swell 110°, '
Depth
(m)
0
h
8
16
26
39
1+9
76
Ikk
21*+
293
373
596
2 ft, 10 sec.
OBSERVED
Temp.
(°C)
29.1+8
25.17
2I+.78
23.96
22.U8
20.95
20.10
16.21
12.85
12.00
11.10
10.00
6.96
(7oo)
33.97
31+.00
31+.02
3^.09
31+.18
31+.28
31+.1+5
31+.72
31+. 87
3l+. 83
3I+.76
31+.70
31+.57
02
(ml/D
1+.56
1+.1+7
1+.53
1+7
11
05
53
0.32
0.1I+
O.09
0.09
O.09
O.09
PO^-P
(ugm at/l)
0.62
0.65
O.67
O.78
0.97
1.10
1.1+1
1.86
I.85
1.92
2.00
2.15
2.38
NOo-N
(ugm at/l)
0.1
0.2
0.2
0.2
0.1+
0.5
0.2
0.2
0.0
0.0
1.6
0.8
0.1+
PH
8.21
8.20
8.19
8.18
8.13
8.06
7.91+
7.85
7.85
7.82
7.80
7-77
7-77
/ Alk
^ millival/l)
2.31
2.31
2.31
2.31
2.31
2.33
2.3I+
2.35
2.35
2.35
2.37
2.38
Depth
(m)
0
10
25
50
75
100
Chlorophyll
"a"
(mg/m )
0.8l6
1.13
0.800
0.529
BIOLOGICAL OBSERVATIONS
Productivity
O.298
Zooplankton Volume:
Bacteria
( no /ml )
75
13
13
58
37
198
in situ
:3_
B-
(mg C/nrYday)
Incubator
(mg C/nr/hr.)
3-5
2.8
0.32
0.15
0.031+
233 ml/1000 m3 total, 233 ml/1000 m3 small.
SUBMARINE DAYLIGHT (I+80 mu)
Read.
Depth
(m)
2
7
12
22
32
1+2
Corr. Sub.
(ua)
759
506
282
102
1+5.6
20.1+
k/m
.0810
.116
.101
.0805
.080l+
7»T/m
92.2
88.9
90.3
92.3
92.3
U-
P-
- 38 -
Station 23
M/V Stranger; SCOPE;
wire angle, 0°; temp.
sea, 2; swell, 310°, i
Depth
(m)
10
30
35
52
63
75
120
197
291
390
1+85
737
Temp.
(°C)
29.04
28.29
28.1+6
28.1*6
24.07
20.97
18. 61+
13.61+
12.06
10.98
9.68
8.28
6.05
34.05
34. 06
34.24
34.33
34.34
34.54
34.54
34.85
3I+.83
3^.76
34.69
34.61
3^.56
December 9, 1956; 2019 GCT; l6°52.0'N, 103'06.0!W; 1580 fm;
8l.2°F dry, 76.8*F wet; weather, 02; Clouds, 1, amt.l;
ft, 6 sec.
(ml/D
4.42
4.34
4.45
4.4o
3.90
1.98
0.89
0.13
0.09
0.09
0.10
0.11
0.10
OBSERVED
PO^-P
NO2-N
PH
Alk
ugm at/l)
(ugm at/l)
(millival/l)
O.38
tr.
8.25
2.31
0.1+0
0.0
8.23
2.30
O.38
0.0
8.21+
2.31
0.39
0.0
8.23
2.32
0.62
0.5
8.16
2.32
1.19
0.2
8.02
2.32
1.58
0.1
7.90
2.33
1.92
0.9
7-79
2.33
1.91
1.6
7-79
2.35
1.99
1.1+
7-75
2.35
2.16
1.0
7.74
2.37
2.26
0.8
7.75
2.37
2.32
tr.
7.72
2.38
Depth Chlorophyll
(m)
0
10
25
50
75
100
3
(mg/m )
0.109
0.11+8
0.161+
0.385
o.i+oo
Zooplankton Volume:
Depth
(m)
2
7
12
22
32
1+2
52
62
72
82
BIOLOGICAL OBSERVATIONS
Productivity
Bacteria
( no /ml )
38
66
65
23
65
51
in situ Incubator
Xmg C/nr/day) (mg C/m^/hr.)
1.8 0.48
0.28
0.15
0.25
O.067
58 ml/lOOOm3 total, 33 ml/lOOOm3 small
SUBMARINE DAYLIGHT (480 mu)
Corr. Sub. Read,
(ua)
745
620
510
368
270
181
102
42.5
19.0
11.6
k/m
.0367
.0391
.0326
.0306
.0404
.0574
.0875
.0805
.0451
B-
V. T/i.
96.4
96.2
96.8
96.9
96.0
94.4
91.6
92.3
95.6
u-
39
Station 24
M/V Stranger; SCOPE; December 10, 1956; 1826 GCT; 19°30.0*N, 105°52.0'W; 2050 fm;
wire angle, 5°; wind, 040°, force 4; temp., 79-8°F dry, T6.30F wet; weather, 03;
clouds, 6, amt. 5; sea, 3; swell, 350°, 10 ft; 7 sec.
OBSERVED
Depth
Temp.
S
°2
P01+-P
N02-N
PH
Alk
(m)
CO
('/..)
(ml/1) (
igm at/l)
(|igm at/l)
(millival/l)
0
26.92
3^.65
4.45
0.39
tr.
-
2.34
6
26.94
34.86
4.46
0.4l
0.0
-
2.31
18
26.88
3^.65
4.44
0.40
0.0
-
2.32
24
26.46
34.77
4.36
0.45
0.0
-
2.32
30
22.38
3^.36
4.34
0.66
tr.
-
2.30
^3
18.99
34.42
1.88
1.30
0.7
-
2.29
80
lk.dk
34.74
0.09
1.84
0.0
-
2.31
131
12.80
34.84
0.12
1.88
2.3
-
2.32
196
11.80
3^.80
0.12
1.92
2.5
-
2.32
290
IO.63
34.74
0.13
2.06
1.8
-
2.33
388
9-4l
3^.67
0.09
2.14
0.8
-
2.33
484
8.08
34.60
0.10
2.25
0.2
-
2.34
733
5.91
3^.58
0.09
2.36
0.0
-
-
BIOLOGICAL OBSERVATIONS
Productivity
Depth
Chlorophyll
"a" ^ "
Bact
sria
in situ
Incubator
(m)
(mg/m )
(no/i
nl) (mg
C/m3/day)
0
(mg C/m /hi
•)
0
0.139
24
2.1
0.59
10
O.I76
19
-
0.4l
25
0.239
12
-
0.53
50
0.523
19
-
0.12
75
-
30
-
-
100
0.254
9
-
0.18
Zooplankton Volume
: 77 ml/lOOOm3 total
, 77 ml/lOOOm small.
SUBMARINE DAYLIGHT
(480 mji)
Depth
(
?orr . Sub .
Read.
(m)
(na)
k/m
/oT/m
2
773
7
656
0.0394
96.1
12
520
0.0465
95.4
22
340
0.0425
95.8
32
155
O.0786
92.4
40 -
Station 25A
M/V Stranger; SCOPE; December 12, 1956; 2108 GCT; 23°31.0'N, 111*22. O'W; 270 fm;
wire angle, 3°; wind, 340°, force 3; temp., 70.8°F dry, 65.0°F wet; weather, 02;
clouds, 0; sea, 3> swell, 370°, 3 ft.
OBSERVED
Depth
Temp.
S
02
POl^-P
N02-N pH
Alk
(m)
(°c)
("/..)
(ml/1)
(ngm at/l)
(ngm at/l)
(millival/l)
0
23.82
34.70
4.46
0.46
0.0
2.36
5
23.76
34.70
4.70
O.38
tr.
2.35
15
23.73
34. 66
4.67
0.40
0.0
2.35
30
23-54
34. 68
4.66
O.38
0.0
2.34
45
23.119
3^.70
4.53
0.37
tr.
2.34
53
,22.38
^22.48
34.59
4.4l
0.50
0.3
2.34
82
15-34
33.95
3.88
0.82
0.1
2.29
142
13.23
34.61
0.39
1.79
tr.
2.33
196
11.92
34.71
0.15
I.83
0.0
2.34
242
11.38
34.72
0.10
1.86
tr!
2.34
239
10.56
34.69
0.15
1.88
tr.
2.34
390
8.88
34.59
0.13
1.96
0.0
2.34
1+86
7.80
34.54
0.12
2.02
tr.
2.35
BIOLOGICAL OBSERVATIONS
Productivity
Depth
Chlorophyll
"a"
Bacteria
in situ
Incubator
B- U- F
(m)
(mg/m
)
( no /ml )
(mg c/m3/day)
(mg c/m3/hr.)
0
Lost
-
6.1
1.2
_
10
0.557
-
3-3
0.62
_
20
-
-
4.9
-
_
25
Lost
-
-
-
_
30
-
-
5.4
-
_
40
-
-
0.96
-
_
50
O.698
-
O.67
0.093
_
70
-
-
0.29
-
_
90
-
-
-
0.041
_
100
Lost
_
0.29
_
_ _ -
Zooplankton Volume: 57 ml/lOOOnT total, 47 ml/lOOOnr small
Water Column Productivity: O.185 gC/m /day
- 4l
Station 25A
(Cont.)
SUBMARINE
DAYLIGHT
(U80 mu)
Depth
Corr. Sub. :
Read.
(m)
(na)
k/m
2
708
_
7
582
0.0392
12
^36
0.0578
22
253
0.05^
32
ll*6
0.05^9
k2
9^.7
0.0 V33
51
51.0
0.0688
61
21.2
0.0878
71
11.6
O.0603
VoT/m
96.2
9^-7
9^.6
95.8
93A
91.6
9^.1
k2 -
Station 25B
M/V Stranger; SCOPE; December 13, 1956; 0238 GCT; 23°31.5IN, 111°19.0'W; 310 fm;
wind, calm; temp., 72.9°F dry, 65.6°F wet; weather, 02; sea, 1; swell, 300°, 2 ft,
OBSERVED
Depth
Temp.
s
0o
(ml/D
P°l+-P ,
N02-N
(m)
C'c)
(*/..)
(iigm at/l)
(ugm at/l
0
23.62
-
^•73
0.37
-
5
23.62
-
1+.70
0.35
-
10
23.63
-
^.57
O.38
-
11+
23.60
~ *
1+.79
0.37
-
20
23.51
-
1+.77
0.39
-
25
23.50
-
1+.70
O.38
-
30
23 A3
-
1+.63
0.1+2
-
35
-
-
1+.79
0.1+0
-
1+0
23.33
-
V.75
O.38
-
^5
-
-
to75
0.52
-
1+9
-
-
to77
0.1+6
-
53
-
-
^•73
0.51
-
59
-
-
1+.1+5
0.61+
-
73
16.10
-
1+.8I+
0.50
-
98
13.58
-
2.96
1.10
-
Ikk
12.96
-
0.1+2
1.83
-
BIOLOGICAL OBSERVATIONS
Productivity
Depth
Chlorophyll
"a"
Bacteria
in situ
Incubator
(m)
(mg/nT)
(no /ml
)
(mg C/m3/day)
(mg c/m3/b
0
O.I+67
32
7-6
-
10
0.1+37
88
1+.2
-
20
-
16
3-1
-
25
O.U37
-
-
-
30
-
23
7-0
-
1+0
-
28
1.2
-
50
0.795
27
2.1
-
70
-
13
0.23
-
90
-
28
-
-
100
0.219
-
-
-
pH Alk
(millival/l)
B- U- P-
Water Column Productivity: 0.220gmC/m /day
- ^3 -
Station 25B
(Cont. )
SUBMARINE DAYLIGHT (l+80 mi)
Depth
Corr. Sub. Read.
(m)
(na)
k/m
°/oT/m
2
6W
_
_
7
506
0. 01+82
95.3
12
376
0.0591+
91+.2
22
200
0.0621
94.0
32
103
O.O67I+
93-5
in
W.5
0.0793
92.1+
1+7
24.5
0.1138
89.2
58
H.7
0.0672
93.5
69
5.4
0.0761+
92.6
- 1+1+
^
a a
O CO
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H O^
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O cd
O -P
t-J O
EH
rH s--
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cd ^.
,cm
o c p
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•P B
o — »
o -~^
rH >>
P* «
*£
col a
CIO
cd
O " on
O H\
4d ^ B
o p< —
o
OJ
o
OJ
NO
OJ
NO
OJ
on -3-
J- o
m -*
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OJ
no
ON
o
LTN
S>
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pn a
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Cd*-'
<HH ft
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-* en
en en
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en en-=t
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on
en
en
on
J-
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on on
OJ ITN-H- OJ L^-NO onoc— ONonr-HonOHonOJonon on no no cm ir\ t— -* -=t itn oj o no o ON
J- -4- NO NO LTN C^NO NOCOCOONONONOONONOOOon onNO
NONONONONQNONONONONONONONO t^NO NO t — t — C — t — t — C —
1TNVO l^- t— CO rH
c~t~t~t~ t— CO
OJ OJ OJ OJ rH
CO CO CO CO CO
ON ONNO ONNO ON on ON rH ONCO NO-H/OJrHt^-OJOJNONONOJ-
J--4-onCMOJrHrHOOL01lfNLfNLfNLfNLfN-H/-HrJ-OJlfNOJLrN
oooooooooooooooooooooo
nonononononononono ir\LrNifNirNLrNirNLrNLrNirNirNJ--=r en
HHHHHHHMHHHHrlHHrlrlHrlHrlH
rHHrHrHrHrHrHrHrHrHrHrHrHrHrHrHrHrHrHrHrHrH
rH C— ONCO LTN on O C— J- on on f- rH IT\C0 en^t OJ onco CO CO
OOJLfNrHirNOJLPvrH-H/ ononOJOJ H O O lAJ ItNrOrl o
oooooooooooooooooooooo
O 0\<n CO f— [^ NO NO ITN ITN LTN LTN ITN ITN LTN IfN J- -=f rr'or><r,Cp
on OJ OJOJOJCMOJCMCMOJOJOJOJOJOJCMCMCMOJOJOJCM
NO NO NO NO NO
NONONONONONONONONONONONONONONONONO LTN LTN LTN ITN IT\
inirNirNirNirvirNLfALrNLfNif^ifNioirNininirNirN-^-----^''
•k......^^^.,.,^-.^**-*- -o o o o o
COCOCnCO ONONONONONONONONONONONONONrH rH rH rH H
oooooooooooooooooooooo
aa^aaaaaaaaaaaaaaaaaaa
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■^OOOJirNonOonO-H^LTNOOOOO
rH J" NO NO C— CO ONOOOnOJCO rlJ; t~"
rlrlHrlrlHHOIMOIOOHrlH
-3- [— O on oj unco
rH rH OJ OJ O O O
O rH OJ On-* LTN NO ON rH POJ lf\ NO
"-■n-* tTNNO t— CO ONrHHrHrHHHrHrHCMOJOJOJOJ
i i»l',,l''''J.A4.4.A
oooooooooooooooooooooo
OJ
NO J- OJ rH ON rH
O
co f- itnno
J- rH CM LTN
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CM
0 0 o o o o
0
o o o o
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onco co r— no
rlrlrlrlHH
o
o o O O
rH rH rH rH rH rH
rH
rH H r-{ H
NO O LTN ON-d- NO
ir\co oj no o
on CM O -* on rH
-=t-
cm rH -* en
o o o o o o
o
O O 0 O
CM OJ OJ rH r-H rH
<)
O O ON ON
OJ OJ OJ OJ OJ OJ
CM
OJ CM rH rH
NO NO NO NO NO NO
NO NO NO NO NO
LTN LfN ITN LTN LTN ITN
LTN IfN LTN ITN LTN
*\
•V *k ^ ^
rH rH rH H rH H
CM
CM CM OJ OJ
rlHHrlrlrl
rH
rH rH rH rH
>>>>>>
>
> > > >
O O O O O O
o
o o o o
a a a a a a
a
a s a a
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o
o o o o
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LTNCO onNO
O O O H rH rH
O
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rH OJ On-* tf\NO
rH
CM on IfN NO
rlHHrlHH
OJ
OJ 01 OJ OJ
- 45 -
3
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o\
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o
cd
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S-l rH 0
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SURFACE CURRENT VELOCITY AND DIRECT:::.'
(GEK OBSERVATIONS
Lat °N
Long °W
Direc-
Velocity
Lat *N
Long *W
-ir-c-
Velocity
tion
0—
cm/sec .
tior.
0—
en sec.
2i+*i+5'
115° h2'
221+
27.5
:-°45'
- - - 3 po 1
173
60.2
lk° 16'
96*33 '
287
9*.8
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100*1+1'
222
22.
12*1+7'
9V1I+'
332
3^. 2
15°12*
100*58'
219
01+.8
11°17'
9l803'
33^
1+7.6
15°39'
101*31+'
-
65.3
11*05'
900 32 '
016
1+5.2
15° 59'
102*01 '
239
10*53'
90° 05'
031
69.5
i6°i8«
102*26'
190
30.8
10*1+2'
89*31+'
038
78.2
16*1+1 '
102 s
21-
15.6
10°31'
89*01+'
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32.6
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280
12.
10° 19'
88° 32'
138
27.7-questionable
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30.0
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88° 29'
21+7
25.2
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103°59'
309
2: .
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89° 11'
3i*
35.9
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306
17-9
09°26'
88° 1+7'
309
50.7
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01+1+
17.2
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88°22'
288
26.1+
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87° 52'
338
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19-5
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56.8
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86*51'
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29.3
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28.0
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81+* k2>
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36.0
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339
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83°13'
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063
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84° 17'
120
21.3
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132
25.7
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173
35.9
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213
29.5
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86°13'
261
12.1+
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86° i+l'
029
10.5
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87°22'
058
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68.5
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81.1
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80.0
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90*28'
170
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90*55'
273
52.1
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92°30'
35€
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93°l6'
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180
37.0
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151*
1+2.0
10° 28'
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1*3
21.1
10° 36'
9i+°36'
112
15.0
10° 56'
95*00'
::r
51.8
11°15'
95*21+'
156
10.0
il033'
95° 1+8'
186
07.9
- 51
PART 2. SCIENTIFIC REPORTS
- 53 -
POSSIBLE APPLICATION OF A BACTERIAL BIOASSAY
IN PRODUCTIVITY STUDIES
William Belser *
An increasing number of reports in the litera-
ture, demonstrating requirements for growth
factors by various marine algae (Levin, 195^-,
Provasoli and Pintner, 19^3a, Sweeney, 195^) ,
the effects of external nutrilites on feeding
responses (Loomis, 1953, Collier, 1950), and
the possible implication of organic micro-
nutrients in the discontinuous distribution of
marine plants and animals (Lucas, 1939, Prova-
soli, 1956, Margalef, 1956, Wilson, 1956) has
led to the formulation of this program. Many
of these organic materials are required in
extremely small amounts, and might be expect-
ed to be present in sea water in very low con-
centration. Previous attempts to isolate and
characterize some of them have been moderately
successful (Johnston, 1955, Provasoli and
Pintner, 1953^), although somewhat cumbersome.
Relatively high salt concentrations in sea
water preclude direct chromatography of the
organic materials, and require their preisols-
tion, either by absorption or desalting. Deal-
ing with materials present in micrograms-per-
liter quantity presents a formidable task.
With these facts in mind, I have considered
the possibility of establishing a series of
biochemical mutants with a wide spectrum of
nutritional requirements, which might be
employed directly as bioassay organisms in
sea water. Attempts to train Escherichia
coli, in which many mutants are already in
culture, to grow in sea water- were time con-
suming and impractical. Therefore, a number
of marine bacteria were screened for desirable
characteristics, and Serratia marinorubrum
(ZoBell, 19^4) was selected as the most suit-
able of these organisms for this purpose.
S. mar i nor ub rum is an easily distinguishable
red pigmented organism, which will grow well
in a medium composed of inorganic salts, with
glycerol as the sole source of carbon. In
addition, it shows a wide range of salt
* Public Health Service Research Fellow of
The National Cancer Institute.
tolerance, growing in media with the salinity
of fresh water, as well as in threefold con-
centrated sea water. This heterosmotic
feature suggests the value of the organism
for bioassay of rivers and lakes, as well as
the ocean, in tide pools, estuaries and
seasonally landlocked sloughs .
To date, several mutants have been obtained
by ultraviolet irradiation of S. marinorubrum.
One of these requires biotin, and will respond
to concentrations in the order of 1 to 5
mug/ml. (see Table l) . The second mutant has
a specific requirement for uracil, and responds
to concentrations between 10 and 100 mug/ml.
The third mutant thus far obtained has a non-
specific purine requirement, and will grow
when supplied with any of the purine bases or
their ribosides. The most sensitive respc:
obtained with this mutant is to hypoxanthi
in the range from 10 to 100 mug/ml.
In a preliminary field trial designed to test
the bioassay system, some 38 sea water samples
were tested. These samples were taken for me
or. SCOPE in waters off the coast of Mexico
and Central America. The results of these
tests shoved quite definitely that the bio-
assay system has merit, since a fairly wide
distribution of biotin was observed, with
sporadic occurrence of uracil, a ' one
instance of purine (see Table 2). Controls
failed to show any evidence of either rever-
sion of any of the mutants or contamination
of the water samples
MATERIALS AND TECHNIQUES
The technique for mutant induction involves
the irradiation of cultures in the logarithmic
phase of growth and screening for mutants
after incubation. This has been done by
minimal enrichment and delayed enrichment
techniques. The mutants, so isolated, are
identified with regard to their specific re-
- 55
TABLE 1
RESPONSE OF MUTANTS TO VARYING CONCENTRATIONS OF THEIR SPECIFIC NUTRILITES
Growth
Requirements
of Mutant
Purine*
Biotin
Uracil
Concentration
for Optimum
Growth
Growth Limiting
Concentration
Range
Lower Limit
Detectable
10 - 2.5
5 - 1.0
5 - 1.0
.7 - -05
.0l+- .002
1.0 - .08
.05
.002
.08
Concentrations expressed in ug/ml.
» Purine response was measured using hypoxanthine as growth factor. The
mutant responds to all four purine bases.
quirements, and their optimal and limiting sub-
strate levels established by the optical den-
sity method of recording growth response. For
use in testing sea-water samples, these are
first filtered through Whatman No. 2 paper to
remove any large particles and then autoclaved.
The water samples are next divided into a num-
ber of replicate samples containing glycerol
as added carbon source, and are then inoculat-
ed with the mutants. Positive tests may be
measured in the Beckman model DU spectrophoto-
meter for quantitative estimation of nutrient
concentration, following a suitable period of
incubation. Each test of unknown sea water is
accompanied by reversion tests on the mutants,
as well as viability controls.
Attempts are in progress to standardize the
technique of testing sea-water samples so that
any technician, either biologist or nonbiolo-
gist, may use it at sea. This would permit
efficient utilization of shipboard facilities
in that a technician in chemistry, for example,
could carry out these tests along with the
chemical work, and leave space for some other
member of the scientific team, by making it
unnecessary for a biological technician to go
along for this purpose.
EXPERIMENTAL RESULTS
The results of the tests on sea-water samples
are presented in Table 2 along with other SCOPE
data to show correlations.
In view of the scant nature of these data and
the small number of samples examined, it would
be presumptuous to attempt to draw any conclu-
sions from this test. The major purpose of the
test was to assess the validity of the bioassay
system, and it seems to indicate that the syste
has merit. There is an indication that
soluble organic materials are present and are
distributed both laterally and vertically in
discontinuous fashion. In comparing the bac-
terial counts and the chlorophyll concentra-
tion with the occurrence of organic material,
it was encouraging to note that presence of
bacteria seems to be inversely correlated,
while there is direct correlation between algal
production and growth factor occurrence.
Whether the algae are present because growth
factors are present, or vice versa, and
whether high bacterial numbers occur at the
expense of external growth factors are problems
which will have to await further experimenta-
tion. A considerable amount of data would
56
TABLE 2
RESULTS OF TESTS OF SEA-WATER SAMPLES WITH THE MUTANTS
Depth
mgm/m
Date of
Depth
Sterility-
Bact .
chloro.
Sample
(meters)
control
b"
u"
p~
count
2
DETN.
Surf.
chlorophyll
H/lO/56
Surface
0
03
0
0
11 an
0.125
11/12/56
Surface
0
0
0
0
-
Surf.
0.204
11/15/56
Surface
0
++
0
0
3^
Surf.
0.577
11/16/56
Surface
0
0
0
0
546
Surf.
O.380
11/21/56
Surface
0
I
0
0
-
Surf.
0.308
11
5
0
T
0
0
-
8
0.310
it
10
0
0
0
0
-
12
0.340
It
Ik
0
T
0
0
187
16
0.342
II
18
0
0
0
0
1+200
20
0.335
II
22
0
-
-
0
383
-
-
II
27
0
6"
6"
0
3200
30
0.458
II
52
0
0
0
0
-
-
-
II
100
0
0
0
0
-
100
0.280
II
200
0
0
0
0
-
200
0.092
12/1/56
Surface
0
0
0
0
75
Surf.
O.329
n
10
0
0
+
++
3
10
0.272
11
50
0
0
+
0
1
50
75
O.491
No RDG
11
75
0
-H-
1
100
0.101
12/2/56
Surface
0
0
0
0
23
Surf.
0.196
11
5
0
-H-
0
0
6
5
11
10
0
-H-
+
0
5
10
0.215
11
25
0
*
+
0
2
25
0.261
11
50
0
-H-
+
0
23(8)
50
0.633
11
75
0
+
0
0
5
75
11
100
0
T
0
0
2
100
0.105
12/3/56
Surface
0
+
0
0
114
Surf.
0.169
12/5/56
Surface
0
-H-
0
0
97
Surf.
0.129
12/7/56
Surface
0
4+
0
0
7
Surf.
ti
25
0
-H-
0
0
l4
25
0.905
1) Growth controls where specific supplement was added were all ++++
Reversion controls were all negative.
2) Glycerol (0.2°/o) added to all samples as carbon source.
growth .
3) 1 l 1 1 = non limiting concentration (optimal growth)
++,+> +, 0 = limiting concentrations; moderate, slight, very slight,
and no growth respectively.
57
have to be gathered and processed before any-
definite conclusions could be drawn, but at
this writing these tests certainly present a
possible approach to an exciting aspect of
primary production in the sea.
BIBLIOGRAPHY
Collier, A., et al. 1950.
A preliminary note on naturally occurring
organic substances in sea water affecting
the feeding of oysters.
Science, Vol. Ill, pp. 151-152.
Johnston R. 1955.
Biologically active compounds in the sea.
Jour. Mar. Biol. Assn. U. K., Vol. 3*4-,
pp. 185-195.
Margalef, R. 1956.
Temporal succession and spatial hetero-
geneity in natural phytoplankton.
Proc. Sym. Perspectives in Mar . Biol, (in
press), U. of Calif. Press.
Provasoli, L. 1956.
Growth factors in marine organisms.
Ibid ( in press) .
Provasoli, L., and I. J. Pintner. 1953b.
Assay of vitamin B, „ in sea water.
Proc. Soc . Protozooiogists, Vol. 4, No. 10.
Sweeney, B. M. 1954.
Gymnodinium splendens, a marine dinoflagell-
Lewin, R. A. 195^.
A marine Stichococcus sp
vitamin B, p (Cobalamin) .
TT41
Jour. Gen.
which requires
crobiol., Vol. 10, pp. 93-96.
Loomis, F. 1953-
Glutathione stimulation of feeding res-
ponse in Hydra.
Unpublished.
Lucas, C. E. 1947-
The ecological effect of external meta-
bolites.
Biol. Rev., Vol. 22, pp. 270-295.
ate requiring vitamin B._.
Am. Jour. Bot, Vol. 4l, pp. 821-821)-.
Wilson, D. P. 1956.
Some problems in larval ecology related
to the localized distribution of bottom
animals.
Proc. Sym. Perspectives in Mar. Biol, (in
press), U. of Calif. Press.
ZoBell, C. E. 19M4-.
A list of marine bacteria including descrip-
tions of sixty new species.
Bull, Scripps Inst. Oceanogr., Vol. 5,
pp. 239-292.
- 58 -
SCOPE MEASUREMENTS OF PRODUCTIVITY, CHLOROPHYLL "a", AND
ZOOPLANKTON VOLUMES
by
R. W. Holmes, M. B. Schaefer, and B. M. Shimada
The productivity, chlorophyll "a", and zooplank-
ton volume data obtained on SCOPE have not yet
been examined in detail. However, some aspects
of sampling variability, and certain of the more
obvious relationships among these quantities,
have been examined and are, in some instances,
compared with similar data and relationships
obtained in 1955 on Eastropic Expedition (Holmes,
Schaefer, and Shimada, 1957) •
SAMPLING VARIABILITY
Ik
Measurements of C uptake and of chloropnyll
"a" are subject to sampling variability due to
the nature of the distribution of phytoplankton
organisms in the sea. The question, therefore,
arises as to how representative of a general
area is a single sample taken from that area.
In order to investigate sampling variability
over a relatively small area, as a first approach
to studying this problem, on November 22nd, 1956
in the vicinity of 09°25' N, 89*31' W, we
collected samples from a grid of nine stations
on a square pattern, the station spacing being
three miles. The station arrangement is shown
in Figure 3- These stations were visited in the
order shown, between 0915 and 1202. At each
station were taken three replicate surface
samples for the determination of C-^ uptake and
a single surface sample for the determination
Of chlorophyll "a".
Ik
C uptake was determined in a 250-ml. aliquot
of each replicate, using 0.9 ^C of C1^, and
incubating each sample for four hours in the
shipboard incubator at the prevailing sea-sur-
face temperature, and at an illumination of
approximately 1000 foot-candles. The incubat-
ed samples were filtered through one-inch-
diameter HA Millipore filters, which were dried
in a desiccator and subsequently counted in a
proportional counter (Nuclear Chicago PC-1).
The counting time was of a duration to give a
total of at least 1000 counts in each instance,
and varied from 6 to 10 minutes. The results
are given in Table 3 in counts per minute, the
uptake (count) being corrected for variations
in light incident in different samples on the
assumption that, over the range of intensity
of illumination employed, the uptake is pro-
portional to the illumination. The error
(standard error) of each determination due to
the statistical variability of counting is in
each case, not over five counts per minute.
For each set of replicate samples, we show in
Table 3 the mean and standard deviation. It
may be observed that the values of standard
deviation are all rather similar, and are not
correlated with the means, except for Station
9-SC-7 where the value of the standard devia-
tion Is very much larger than that at any of
the other stations. The large variation at
this station appears to be due to the single
replicate giving the very high value of 680
cpm. which may be aberrant.
An analysis of variance of the nine sets of
three replicates (Table k) , including the
suspect sample, indicates that the variance
among station means is no greater than could
be expected to occur by chance in the light
of the variability among replicates within
stations. The grand mean of 27 observations
is 306.5 cpm., with a standard deviation of
90.9.
Now it may be seen that the value of 680 cpm.,
deviating by 3fk cpm., from the mean of all
observations, is a deviation of over four
standard deviations from the mean value, and
thus is very unlikely to be a chance event.
It appears that this sample is somehow quite
aberrant and should be discarded from the
analysis. Omitting this sample (Table h) de-
creases all variance components very greatly.
The analysis of variance with this sample
omitted still indicates no difference among
stations that could not be expected by chance
- 59 -
STATION ARRANGEMENT
OF
SAMPLING GRID
9
2
3
•
•
•
&•
1
0
SURFACE
PARACHUTE
BOUY
• 4
a .
~X Ml CO
, a
•
5
• *
7
• •
6
FIGURE 3. Station arrangement In sampling grid
-60 -
TABLE 3
Ik
C UPTAKE IN REPLICATE SAMPLES FROM NINE STATIONS OF SAMPLING GRID
Station
Counts per minute
Mean
Variance
Standard
Replicate
(mean square)
deviation
1
2
3
9-SG-l
210
297
26l
256
1,911
^3-7
9-SG-2
252
308
278
279
785
28.0
9-SG-3
293
275
308
292
273
16.5
9-SG-h
299
217
321
279
3,oo^
5^.8
9-SG-5
357
I+87
322
389
7,558
86.9
9-SG-6
258
32^
229
270
2,375
1+8.7
9-SG-7
289
255
680
l»o8
55,777
236.2
9-SG-8
321
260
310
297
1,052
32. k
9-SG-9
29^4-
28U
287
288
26
5.1
Grand mean
306.5
8,266
90.9
Grand mean ex-
c
luding 9-SG
-7
replicate No
. 3
292
2,791
52.8
61 -
TABLE h
ill.
ANALYSES OF VARIANCE OF C UPTAKE AT STATIONS OF SAMPLING GRID
Source of variation Degree of Sum of Mean Variance
freedom squares square ratio
All observations:
Total 26 2lU,090 8,266
Among stations 8 69,38^ 8,673 1.073
Within stations l8 1^5,525 8,085
Omitting station 9-SG-7
replicate 3:
Total 25 69,781 2,791
Among stations 8 35,232 k,kOk 2.17
Within stations 17 3^,5^9 2,032
- 62
TABLE 5
CHLOROPHYLL "a" CONTENT OF SURFACE SAMPLES TAKEN
AT THE NINE STATIONS OF SAMPLING GRID
Station Chlorophyll "a"
No.
mg/nP
9-SG-l
O.U2I4-
9-SG-2
o.it-29
9-SG-3
0.365
9-SG-4
0.U00
9-SG-5
o.Uo^
9-SG-6
0.33^
9-SG-T
O.kOQ
9-SG-8
0.1+51
9-SG-9
0.463
Mean
0.U1U
Standard deviation
0.0k8
- 63 -
500
400
to
E
O
O
o
6
UJ
_l
O
>
-z.
o
I—
a.
o
o
M
300
200
• o
•• •
• o
100
•o °
#
• :% u e
oo
«• /
o SCOPE
• EASTR0PIC
0.1 0.2 0.3 0.4 0.5 0.6 0.7
SURFACE CHLOROPHYLL a mg/m3
FIGURE 4. The relationship between surface chlorophyll "a"
and zooplankton volume.
0.8
64 -
from the within- station variability. With
the aberrant sample discarded, we have a
grand mean of 292 cpm. with a standard devia-
tion of 52.8 cpm.
There is then, no evidence of heterogeneity
among the nine stations. Any single sample
should give a fair estimate of the productiv-
ity of this 6 mile square area, with, however,
a standard error of 52.8 cpm. which is l8°/0
Of the mean value encountered.
Chlorophyll "a" was determined from a single
6 liter surface sample at each station. The
results are given in Table 5.
Since we have only a single chlorophyll sample
from each station, we cannot examine the ques-
tion of heterogeneity of this constituent among
stations. The degree to which any single sam-
ple may be expected to represent the mean
value within the area may, however, be Judged
from the standard deviation among the nine
samples of O.OhQ mg/m^, which is 12°/0 of the
mean value.
Superimposed upon sampling variability, of
course, would be that due to the inherent
diurnal periodicity in photosynthesis and
chlorophyll (see p. 82). However, such a
periodicity is not evident in these particular
data.
SURFACE CHLOROPHYLL "a" ZOOPLANKTON VOLUME
RELATIONSHIPS
In figure h surface chlorophyll "a" and zoo-
plankton volume data obtained on both SCOPE
and EASTROPIC are illustrated. While there is
some scatter, the observations reveal that a
positive relationship exists between these
two quantities. The correlation might have
been improved had we a sufficient number of
vertical chlorophyll "a" profiles for inte-
gration and comparison with the zooplankton
volumes .
Similar data from mid to high northern and
high southern latitudes have frequently
shown an inverse correlation, or none at
all. The lack of extreme variation in this
relationship indicates that there may exist
in these tropical waters a situation more
closely approaching a steady state condition
than is found in other waters. Furthermore,
the general agreement among surface chloro-
phyll "a", surface productivity, and zooplank-
ton volume shows that any one of these will
serve to indicate the general level of the
other two.
PRODUCTIVITY PER UNIT CHLOROPHYLL "a" IN SUR-
FACE PHYTOPLANKTON
The relationship between photosynthesis and
chlorophyll "a" concentration has been
studied and discussed by a number of investi-
gators - see for instance Glendenning et al.
(1956), Rabinowitch (1956), Ryther (l955)~
Few of these observations include data for
marine phytoplankton species, but it is of
interest to note that for those studied the
ratios are quite similar to those observed in
some land plants and algae.
The SCOPE data available for this comparison
are of two types: a) the photosynthetic
rate obtained with surface samples inoculated
with C14' and incubated for four hours under
constant light (1000 ±. ItO foot-candles) at
approximately the temperature of the sea
surface, and b) the rate based on Cll+ inoculat-
ed samples trailed astern of the vessel at the
surface for about six hours (sunrise to noon,
and noon to sunset). Chlorophyll "a" deter-
minations were made with water samples col-
lected at, or nearly at the same time, as
the samples for the photosynthesis studies.
lit-
The amount of C fixed per hour at 1000 foot-
candles in surface water samples is presented
as a function of surface chlorophyll "a" concen-
tration at each station in figure 5. While
there is considerable scatter, the data appear
to fall into two discrete clusters. Best-fit
lines "drawn" by eye through these two groups
yield the following rates: 7.3 and 2.5 mg
C/hr/mg chlorophyll "a", or, 26.8 and 9.2 mg
C02/hr/mg chlorophyll "a", respectively.
Ryther and Gertsch (1957) give an average
value of 3.7 mg C/hr/mg chlorophyll "a" for
natural populations at 1500 foot candles. This
65 -
5.5
O
o
o
°l
a
_ 2.0
-c
10
E
3 1.5
IE
O
< 1.0
o
3
<_>
-i 1 1 1 1 1 1 1 1—| 1 1 1 1 1 1 1 1 r
8 o
SCOPE 0
EASTROPIC a
Jl I I I I I i I i | |
0 1 0 2 0 3 04 0 5 0 6 0 7 0.8 09 10
CHLOROPHYLL a mg/m3
FIGURE 5. The relationship between surface clorophyll "a"
and incubator production.
- 66 -
3C
25
o
TO
o
E 20
Q
o
cr
Q_
uj 15
o
<
U-
<r
3
in
Z>
H
10
t — i — r
1 — i — i — i — i — i — i — i — i — i — i — i — i — r
o
• CO
• O o
o o
0°
J_2_L
o
I I I
o SCOPE
• EASTROPIC
i i i i i i i i i i
0 01 02 03 0 4 05 06 0.7 0.8 0.9 1.0
CHLOROPHYLL a mg/m3
FIGURE 6. The relationship between surface chlorophyll "a"
and in situ primary production.
-67 -
latter value Is very similar to the average value
reported above for the group exhibiting the low
assimilation factor when the difference in light
intensities between the two sets of experiments
are equated. Tailing (personal communication)
has observed similar rates in cultures of the
marine diatom, Chaetoceros aff inis.
The high values (ca. 20 mg C/hr/mg chlorophyll
"a") observed at a few stations are somewhat
anomalous and difficult to interpret. The sta-
tions where such high values were observed were
all located in the region of the thermal anti-
cline. These higher photosynthetic rates may
be associated with differences in the species
composition and/or some difference in the physio-
logical state of the organisms.
The "average" in situ rates (see Fig. 6) observed
on the expedition on a per-hour basis (assuming
a 12-hour day) are somewhat lower than the rates
observed in the incubator, averaging about U.2
mgC/hr/mg chlorophyll "a". Again, the highest
ratios are observed at stations in the region
of the thermal anticline off Costa Rica. The
lower "average" value may be the result of in-
hibition of photosynthesis during the brightest
portion of the day.
As mentioned above, there is considerable vari-
ability among individual values. Some of the
variability is doubtless associated with the
techniques employed but much of it is certainly
quite real. A study of the possible effect of
differences in species composition of the phy-
toplankton on the productivity-chlorophyll ratio
will be examined in the near future.
These data yield further confirmation of Ryther's
suggestions (1956) that it should be possible to
estimate productivity from the concentration of
chlorophyll "a" in sea water. However, the pre-
cision of such an estimate from chlorophyll "a"
concentration would be rather poor in the Eastern
Tropical Pacific. Furthermore, it appears that
departures from the "average" value of carbon
assimilation per unit chlorophyll "a" that is
observed in the surface water become more pronounc-
ed in samples collected deeper in the photic zone.
BIBLIOGRAPHY
Clendenning, K. A., T. E. Brown, and
H. C. Eyster. 1956.
Comparative studies of photosynthesis in
Nostoc muscorum and Chlorella pyrenoidosa.
Can. J. Bot., Vol. 3^, PP- 9^3-966.
Holmes, R. W., M. B. Schaefer, and
B. M. Shimada. 1957.
Primary production, chlorophyll, and
zooplankton volumes in the Eastern
Tropical Pacific Ocean. Inter-Am. Trop.
Tuna Comm. , Bull., Vol.2, No. h.
Manning, W. H., and R. E. Juday. 1951.
The chlorophyll content and productivity
of some lakes in northern Wisconsin.
Trans. Wis. Acd. Sci., Arts, and Let.,
Vol. 33, PP. 363-393.
Rabinowitch, E. I. 1956.
Photosynthesis and related processes
Vol. 2, Part 2, Kinetics of photo-
synthesis, pp. 1211-2088, Interscience
Publishers, N. Y.
Ryther, J. H. 1956.
The measurement of primary production.
Limm. and Ocean., Vol. 1, No. 2, pp. 79-93-
Ryther, J. H., and C. S. Yentsch.
The estimation of phytoplankton produc-
tion in the ocean from chlorophyll and
light data. Limn, and Ocean., Vol. 2,
No. 3, PP- 281-286.
Steemann Nielsen, E. 1952. ^
The use of radio-active carbon (C )
for measuring the organic production in
the sea. J. du Conseil., Vol. l8, No. 2,
pp. 117-11*0.
- 68
SIZE FRACTIONATION OF PHOTOSYNTHESIZING PHYTOPLANKTON
by
Robert W. Holmes
To estimate the size ranges of photosynthesiz-
ing phytoplankton in tropical waters, three
simple experiments were performed on SCOPE.
In each experiment a surface water sample of
0.5 or 1.0 liter was inoculated with approxi-
mately 20 uc of C1 and placed just below the
sea surface for incubation. After 2-3 hours'
incubation an aliquot was taken from the sam-
ple and passed through a series of filters in
the following order: a disk of No. 20 bolting
silk (mesh size 106u x 106u by measurement), a
disk of nylon bolting material (mesh size 30u
x 3Cu by measurement), an AA Millipore filter
(pore size specified by the manufacturer as
0.8u t 0.05m), and lastly an HA Millipore fil-
ter (pore size likewise specified as 0.1*5u 1
0.02u). In addition in 2 experiments another
aliquot was filtered directly through an HA
Millipore filter. The pieces of netting and
filters were dried and counted in the usual
manner (see p. ?)• The results of these ex-
periments are given in Table 6.
From these data it can be readily seen that
the activity of organisms retained by the
bolting silk and nylon bolting material rep-
resented a small fraction of the total activ-
ity. In two out of three experiments only
about one-half of the total activity was re-
tained by the AA Millipore filter.
It is difficult to believe that all of the
activity passed by the AA Millipore filter was
contained in bacterial cells. Dark-bottle
C fixation in experiments of 6 hours' dura-
tion in these same waters usually averaged
10°/o and never exceeded l8°/0 of the light-
bottle uptake. It seems more plausible to
suggest two alternative explanations. Extreme-
ly small photosynthesizing organisms (less than
about lu) may have been present in these waters
and passed through the AA filter and/or the
bulk of material passed by the AA Millipore
filter may have been cell fragments produced by
the rupture and disintegration of some of the
cells as they impinged upon the membrane-filter
surface during filtration. Unfortunately the
water samples collected for the purpose of
flagellate enumeration and identification have
not yet been examined carefully but it appears
from a cursory examination that the smallest
naked flagellate visible in these samples are
between 1-1.5 l-i in "diameter." Organisms
smaller than those observed in the fixed mater-
ial may exist in the sea but may not have been
preserved adequately enough to permit enumera-
tion or identification. However, it seems un-
likely that a significant portion of the total
photosynthesizing biomass could have been such
organisms .
It seems more plausible that the material pass-
ing through the AA millipore filter was large-
ly in the form of protoplasmic fragments
released from fragile cells which ruptured on
the filter surface. That small naked flagel-
lates do disintegrate as a result of filtration
has been observed by the author and by Dr.
W. Rodhe (personal communication) by comparing
the flagellate abundances on cleared Millipore
filters with those in unfiltered samples. Con-
firmation of this fragmentation hypothesis
has also been observed by the author and Dr.
R. Lasker (unpublished results) who used
radioactive bacteria-free cultures of
C hi amy domo na s sp. Of nine aliquot s, three
were filtered through AA Millipore filters,
three through HA, and three through PH. No
essential. difference in the activities of the
HA and PH filter membranes was observed where-
as the activity in the AA Millipore filter
averaged 12-19°/o less than that observed on
the HA or PH filters. The Chlamydomonas em-
ployed in this study was quite healthy and the
cells averaged about 8u in "diameter." A some-
what greater difference was observed in another
experiment when the filtrate of a nonbacteria-
free culture from the AA filter, was passed
successively through an HA and PH filter
here the AA filter passed about 32°/0 of the
activity retained by all three filters. This
apparent difference in retention is probably
the result of fragmentation caused by the fil-
tration through the AA Millipore filter and
the passage of some bacteria less than 0.8 \i
in size.
69
TABLE 6
Exp.
// 1 Nov. 20, 1956
No.
Filtration procedure
Filter
activity
c/m
°/o Of
totnl
activity
-
500 mis filtered through HA Millipore
1058
1
2
3
-
500 mis " " No.
500 mis of filtrate from No.
n 11 if » 11 jyQ
" " " " " No,
20 bolting silk
2 through nylon
3 AA Millipore
h EA Millipore
k2
32
U78
596
3.7
2.8
in. 6
51.9
// 2 Dec.
2, 1956
TOTAL
11^
100
Exp.
No.
Filtration procedure
Filter
activity
c/m
°/o Of
total
activity
1 250 mis filtered through No. 20 bolting silk 8 0.5
2 250 mis of filtrate from No. 1 filtered through
nylon l8 1.1
3 250 " " " from No. 2 " through
AA Millipore 1U38 90.
h 250 " " from No. 3 filtered through
HA Millipore 13^ &.h
TOTAL
1598
100
Exp. // 3
No,
Filtration procedure
Filter "To oT~
activity total
c/m activity
250 mis filtered through HA Millipore
117
1
2
3
1+
I! If
No. 20 bolting silk
250 mis of filtrate from No. 2 filtered through
nylon
No. 3 " through
AA Millipore
No. k " through
___^ HA Millipore
n 11 11
11 11 it
0
0
71
57
0
0
55.5
1^.5
TOTAL
128
100
70
These observations are apparently at variance
with those reported by Steemann Nielsen (1952).
The experiment designed by Steemann Nielsen
(1952) is difficult to interpret because all of
the necessary information is not given. Never-
theless, it appears that Steemann Nielsen fil-
tered aliquots of tropical surface phytoplank-
ton through filters of varying porosity, the
coarsest having a maximum pore size slightly
in excess of 1 u. In these two experiments
no difference in retention was observed be-
tween the various filters and Steemann Nielsen
concluded that all important autotropic organ-
Isms in these samples were larger than 1 u.
It would seem from the SCOPE experiments that
the conclusion of Steemann Nielsen cannot be ap-
plied universally. In two of the SCOPE experi-
ments only about half of the radioactive mat-
erial was retained on the AA filter, with pore
size of 0.8 u. There can be little doubt that
the amount of material retained on filters of
this porosity will vary with the population
composition and perhaps its physiological con-
dition. While the bulk of the photosynthesiz-
ing biomass appears to be in the size range of
1-30 u, if an assessment of the total activity
in a water sample is desired, it would seem
advisable to employ filters with a maximum
pore size somewhat less than 0.5 (!•
BIBLIOGRAPHY"
Steemann Nielsen, E. .^
The use of radio-active carbon (C )
for measuring organic production in
the sea.
J. du Cons., Vol. 28, No. 2, pp.ll7-ll*0,
1952.
71
DIURNAL VARIATION IN THE PHOTOSYNTHESIS OF NATURAL
PHYTOPLANKTON POPULATIONS IN ARTIFICIAL LIGHT
by
Robert W. Holmes and Francis T. Haxo
Evidence for the existence of a daily period-
icity in photosynthesis of marine phytoplank-
ton has been presented In two recent papers
(Doty and Oguri, 1957> and Yentsch and Ryther,
1957) • These authors observed a diurnal per-
iodicity In photosynthesis in surface-water
samples collected at intervals throughout the
night and day, illuminated under constant
light, and kept at a constant temperature.
The rate of photosynthesis in the water samples
began to increase during the early morning
hours and reached a maximum at about 0800 hours .
This was followed by a rapid decrease. At
about 1800 hours a low level of photosynthesis
was reached and was maintained until about 2^00
hours when the predawn rise began culminating
in the 0800 maximum.
Two preliminary experiments are described be-
low which were designed to study this rhythm
in the eastern Pacific. The techniques em-
ployed were similar to those of Doty and Oguri
(1957). Surface samples were collected at
two-hour intervals (three-hour intervals during
the second experiment) alongside a free-float-
ing surface buoy to which was attached, just
below the sea surface, a regulation U. S. Navy
parachute. The samples were collected in a
large plastic bucket and two 250-ml. aliquot s
immediately inoculated with 1 uc of C^- and
placed in the shipboard incubator . These
samples were subjected to constant illumina-
tion (about 1000 foot-candles) from daylight-
type fluorescent lights and kept at a tempera-
ture slightly exceeding (about 1°C) the sea-sur-
face temperature for approximately two hours.
After the incubation period the samples were
filtered through 1-in. HA Millipore filters.
The filters were then dried and counted in the
normal manner (see p. 7)- The data were
corrected for any slight deviation in the dura-
tion of the incubation period. In the second
experiment, dark-bottle uptake was subtracted
from the uptake in the illuminated bottles.
The results of these two experiments are Illus-
trated in figures 7 and 8.
The results of both experiments clearly indicate
that the photosynthesis of samples collected
between 1800 and 0200 hours was less than that
observed during the remainder of the 2k-hour
period. The difference between the maximum
and minimum uptake varied by a factor of 5-8.
This Is somewhat less extreme than that ob-
served by Doty and Oguri (1957) and greater
than that reported by Yentsch and Ryther (1957K
In the first experiment (Fig. 7) the daily
maximum occurred between 0800 and 1000 hours
while in the second experiment (Fig. 8) a max-
imum was observed between 1200 and llK)0 hours .
Unfortunately, the 0900 samples were lost in
this second experiment for the daily maximum
might have occurred at about this time.
The time Of the photosynthesis maximum cannot
be defined with certainty since the C^
measurements were discontinuous, representing
averaged rates of uptake for 2-3 hour incuba-
tion periods of samples collected at 2-3 hour
intervals . The data show that the photosyn-
thetic activity of surface waters varies-
diurnally. Such a periodicity may be associat-
ed with concomitant changes in phytoplankton
standing crop or may be a manifestation of an
inherent photosynthetic rhythm. Samples
collected in Experiment No. 1 to assess the
first of these possibilities have not yet
been examined. In the second experiment
chlorophyll "a" determinations made at the
beginning of each 3-hour incubation period in-
dicated a fairly constant chlorophyll "a"
content. These preliminary results differ
from those of Yentsch and Ryther (1957) and
B. M. Shimada (personal communication) who
have observed a diurnal periodicity in
chlorophyll "a" quite similar to the photo-
synthetic periodicity.
73
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I
EXPERIMENT NO. 2
DEC. 5 -6 ,1956
07° 52'N -91° 19'W
0000
0200
0300 0600 0900 1200 1500 1800
0500 0800 1100 1400 1700 2000
TIME AND LENGTH OF INCUBATION
2100
2300
FIGURE 8. Diurnal variations in incubator productivity and
chlorophyll "a" at 07° 52'N, 91°19'W.
-75 -
BIBLIOGRAPHY
Doty, M. S., and M. Oguri. 1957-
Evidence for a photosynthetic daily
periodicity.
Limm. and Ocean., Vol. 2, No.l, pp.37-40.
Yentsch, C. S., and J. H. Ryther, 1957.
Short-term variationsin phytoplankton
chlorophyll and their significance. Limm
and Ocean., Vol. 2, No. 2, pp. ll40-l42.
- 76 -
ATTACHMENT OF MARINE BACTERIA TO ZOOPLANKTON
by
Galen E. Jones
The abundance of marine bacteria living free in
the open sea is low (ZoBell, 191*6) . The reasons
for the small numbers of microorganisms in the
sea have been considered by various workers as
summarized by Orlob (1956). Dilute amounts of
organic matter on solid surfaces cause marine
bacteria to concentrate on these surfaces (Stark
et al., 1938; Heukelekian and Heller, 191*0).
In addition, many marine bacteria demonstrate
definite attachment propensities (ZoBell, 191*6) .
If marine bacteria attach to the surface of liv-
ing organisms there is the opportunity for a
symbiosis between the bacteria and the phytop-
lankton or zooplankton whereby the metabolic
products from both groups might benefit each
other 5 commensalism whereby one of the members
of the association is benefited; or antagonism
where one or both members may be inhibited by
the products of the other.
This investigation was conducted to obtain informa-
tion concerning the numbers of bacteria attached
to plankton as opposed to those living free in
the water.
METHODS
A -meter-net tow was taken at a depth of 20 m.
with a new, clean net at 08°55' N latitude,
88°1*7' W longitude. One species of zooplank-
ton predominated: the red radiolarian,
Castanidium cf . longispinum Haecker. Few
other zooplankters existed in this sample.
Immediately after the net was pulled aboard, about
one g.wet weight of these packed radiolarians
was transferred to a prescription bottle contain-
ing 1*5 ml. of sea water. Another sample of the
radiolarians was transferred to a bottle con-
taining 10 ppm. of the surface active agent,
Tween 80, In 1*5 ml. of sea water. Tween 80, a
relatively nontoxic surface active agent for
marine bacteria (Jones, 1957) was used in an
effort to remove bacteria from the plankton.
It was estimated that the wet weight of the scoop
of radiolarians placed in the Tween 80 was about
two-thirds of that in the sterile sea water.
Both of these samples were diluted l/lOO with
sterile sea water. These samples were shaken
vigorously for one minute.
RESULTS
Inocula were taken from l/lOO dilution as
follows: 0.1, 0.5 and 1.0 ml. These were
plated by the pour-plate technique into a
peptone-yeast extract medium (Oppenheimer
and ZoBell, 1952). The plates were Incubated
for 11 days at 29-31°C. The results appear
in Table 7.
Little quantitative information can be
derived from this particular experiment
since the inoculum was not weighed and the
amounts of plankton in the bottles were not
estimated as equal. If the estimate of two-
thirds as many radiolarians in the bottle
shaken with 10 ppm. Tween 80 can be assumed
correct, the Tween 80 had little effect on
dislodging additional bacteria from the zoo-
plankters. However, the number of bacteria
associated with the radiolarians was certainly
considerably higher than exist free in the
water as estimated by other bacterial counts
recorded in this cruise. It can be assumed
from this experiment that there are between
50,000 and 100,000 bacteria per gram of wet
radiolarians which is about 10-^more bacteria
than are generally present in sea water.
This experiment should be taken only as in-
dicative of attachment of bacteria to
marine plankton since only one group of
organisms, was tested because the actual
numbers of bacteria free in the water was
not measured at the same place and time.
However, the order of magnitude of bacteria
found associated with the radiolarian,
Castanidium cf . longispinum Haecker, when
compared with the bacteria generally found
in similar waters strongly suggests intimate
association.
ACKNOWLEDGEMENT
The author would like to thank Mr. William
R. Riedel, Scripps Institution of Oceano-
graphy, for his identification of the radio-
larian, Castanidium cf . longispinum Haecker.
77 -
TABLE 7
BACTERIAL COUNTS FROM WET PACKS OF RADIOLARIANS (0.5
to 1.0 g.) COLLECTED AT 08°55' N LATITUDE, 88°l+7" W
LONGITUDE:
Dilution Shaken with no Tveen 80 Shaken with 10 ppm Tween 80
Plate count Bacteria/ml Plate count Bacterial/ml
1:1+5000 3 135,000 2 90,000
1:9000 3 27,000 1 9,000
1:1+500 12 5^,000 10 1+5,000
Average 72,000 1+8,000
BIBLIOGRAPHY
Heukelekian, H., and A. Heller. I9I+O.
Relation between food concentration and
surface for bacterial grovth.
Jour. Bacterid., Vol. 1+0, pp. 51+7-558.
Jones, G. E. 1957.
The effects of organic metabolites on
the development of marine bacteria.
Bacteriol. Proc, pp. 16.
Oppenheimer, C. H., and C. E. ZoBell. 1952.
The growth and viability of sixty-three
species of marine bacteria as influenced
by hydrostatic pressure.
Jour. Mar. Res., Vol. 11, No. 1, pp. 10-18.
Or lob, G. T. 1956.
Viability of sewage bacteria in sea water.
Sewage and Industrial Wastes, Vol. 28, No.
9, PP. 111+7-1167.
Stark, W. E., Janice Stadler, and Elizabeth McCoy.
1938. Some factors affecting the bac-
terial population of freshwater lakes.
Jour. Bacterid., Vol. 36, pp.653-65l+.
ZoBell, C. E. 191+6.
Marine Microbiology: A Monograph on Hydro-
bacteriology.
Chronica Botanica Co., Waltham, Mass.
- 78 -
PRELIMINARY STUDIES OF BACTERIAL GROWTH IN RELATION TO DARK AND LIGHT
FIXATION OF CO- DURING PRODUCTIVITY DETERMINATIONS
G. E. Jones, W. H. Thomas, and F. T. Haxo
The technique of studying, productivity hy use
of radioactive carbon (C ) has been widely
employed in recent years (Steemann Nielsen,
1951, 1952, 1951*-; Ryther and Vaccaro, 195k;
Ryther, 1956b). Steemann Nielsen (1952), us-
ing the green alga, Scenedesmus quadricauda,
showed that C^ assimilation in the dark was
very small (about l*/» of maximal fixation in
the light). When this method is used in a
natural ecosystem, such as in the pelagic
waters of the open ocean, a complex of factors
and organisms must be considered. Both phytop-
lankton and zooplankton assimilate carbon
dioxide in the dark. Chemosynthetic bacteria
fix carbon dioxide as their sole source of
carbon and even heterotrophic bacteria fix
some of their carbon as carbon dioxide via
the Wood-Werkman reaction (Wood and Werkman,
1938, 19IK); Wood et al., 19^1; and Utter and
Wood, 1951). Consequently, mixed populations
collected from nature might be expected to fix
a greater percentage of CO2 in the dark than
the 1% reported by Steemann Nielsen (1952).
The complicating effects of the presence of
bacteria have been recognized, although as yet
unsatisfactorily assessed, in productivity
measurements.
One of the first methods of estimating product-
ivity was the "light and dark bottle" experiments
of Gaarder and Gran (1927). When this technique
is used, water samples from various depths are
dispensed in bottles and lowered to different
depths for time intervals of a day or more.
Oxygen production in dark (wrapped with masking
tape to prevent the entrance of light) and
light bottles is determined by measuring the
quantities of oxygen before and after the in-
cubation period. A modification of this method
was used in the productivity measurements of
Riley (1938, 1939, 19^1a, 191+lb). Riley's
rather large estimates for productivity in the
Sargasso Sea were criticized by Steemann Nielsen
on the grounds that the bactericidal effect of
sunlight inhibited the bacteria in the light
bottles, causing a considerable difference in
oxygen content between the light and dark
bottles (Steemann Nielsen, 1952). It was
correctly pointed out by Steemann Nielsen
that the added surface of the containers
would promote bacterial growth to a far greater
degree than in pelagic sea water under natural
conditions in both light and dark bottles
(ZoBell and Anderson, 193^) . However, while
the bacterial activities in the bottles are
increased, most of the wave-lengths shorter
than 3500 A which are most inhibitory toward
bacteria are absorbed by glass bottles (Vaccaro
and Ryther, 195^). Also, those solar radia-
tions transmitted at a depth of 10 inches did
not affect the growth of marine bacteria as com-
pared with bacterial development in the dark.
Ten inches is the depth employed by Riley in
his experiments. Steemann Nielsen later (1955)
presented experiments suggesting that anti-
biotics produced by the plankton algae in
the light decreased the bacterial activity.
An antibiotic from Chlorella, chlorellin,
has been reported (Pratt et al., 19^4).
It was the purpose of the following experiments
to assess the numbers of bacteria developing
in light and dark bottles containing sea-
water samples from the tropical Pacific Ocean
over different periods of time ranging up
to 1+0 hours and estimate their influence on
the carbon dioxide fixed by the total popula-
tion. Any influence of the planktonic pop-
ulation on the marine bacteria was also noted.
METHODS
The 250-ml. glass -stoppered reagent bottles
used in these experiments were cleaned as
follows : thoroughly washed with a detergent
("Tide"), rinsed three or four times with
sea water, filled with 10°/« HC1 for at least
5 to 10 minutes and rinsed five or six times
with sea water. The surface sea-water samples
were collected in a plastic bucket (cleaned
as above) and dispensed into the reagent
bottles. The bottles were always rinsed
with the sea-water sample before filling.
Radioactive NaHCll+Oo (0.9uc) was added
to each bottle. The dark bottles
- 79
were very carefully covered with black tape to
exclude all light. An attempt to obtain dark
bottles by spraying with black paint failed to
produce light-tight bottles since very small
holes in the painted surface permitted light
to pass. The bottles were incubated in an
illuminated water bath (fluorescent lighting
through a glass bottom) in a random distribu-
tion. Illuminance was measured with a Weston
856 YE photocell connected to a 0-100-micro-
ammeter having a 50-ohm internal resistance.
The meter was calibrated against a Weston
model 756 laboratory illumination meter.
Illuminance measured at the glass bottom of
the water bath was 1100 to 1550 foot-candles.
The average illuminance in the bottles was
about 8o"/0 of this figure. This average
illuminance was about 33°/o of saturation if
saturation illuminance is taken to be 3200
foot-candles (Steemann Nielsen, 1952), or was
53°/o of saturation if Ryther's (1956) average
value for 1*4- different phytoplankton species
(2000 foot-candles) is used.
Just before the bottles were placed in the
water bath (zero hour) and after each in-
terval of time, an appropriate aliquot of the
sea-water sample was removed with a sterile
pipette and plated in duplicate for each
bottle in sterile plastic petri dishes on a
peptone-yeast extract agar (Oppenheimer and
ZoBell, 1952). The bottles were shaken
thoroughly before the sample was removed for
the pour-plate determination. The plates
were poured with agar at k2 _ 2°C on a sus-
pended table which was steadied in moderate
seas with the aid of another person. One
person could operate the suspended table in a
calm sea, whereas pouring plates was imposs-
ible in a heavy sea. The plates were incubated
in the dark at 31 ± 1°C for three days and
then examined with a Quebec colony counter for
the heterotrophic marine bacterial count.
The water sample from each bottle was filtered
through a Millipore HA filter (0.k5 ± 0.02 micra
Millipore Filter Corporation, Watertown, Mass.)
which retained all of the plankton and most of
the bacteria in the samples. The filters were
washed with nonradioactive sea water, dried in
a desiccator over silica gel, and the radio-
active count determined in a proportional flow
counter (Nuclear Measurement Corporation, PC-1) .
RESULTS
Experiments were conducted to determine the
increase in bacterial numbers, and Cl^
assimilation over an l8-hour period in
bottles that were cleaned and in bottles
that were cleaned and sterilized. Surface
sea water was collected at 17°5^' N latitude,
103o50' W longitude (BT Station 3-5) and
incubated in light bottles for 0, 1, 2, k,
8, and l8 hours at 30 t 1*C (sea-surface
temperature, 28.8°C) in the illuminated (1250
j| 150 foot-candles) water bath. The results
of this experiment appear in Table 8.
The data from a similar experiment for surface
sea water collected at ll°4l' N latitude,
91°52' W longitude (BT Station 7 - 5), in-
cubated under identical conditions for 0, 2,
h, 6, 8, 13, and 18 hours is presented in
Table 9.
Ik
While C assimilation was slightly higher in
the autoc laved bottles throughout most of
the experimental period, this difference is
not significant. Bacterial growth was lower
in the autoclaved bottles in the early parts
of the experiments (up to 13 hours), but was
greater at 18 hours. The reason for the
higher bacterial population in autoclaved
bottles at the end of the experiment is not
clear, but may be due to release of nutrients
by autoclaving the bacteria originally present,
or to possible "antibiotic" activities of the
original bacteria. However, autoclaving does
not appear to be necessary in carrying out
a production determination, since the
C-*- values are not significantly different
during the customary experimental period
(8 hours or less) .
An experiment to determine bacterial increases
and carbon dioxide assimilation for a more
prolonged period using the C^ fixaction
method in both light and dark bottles was
, carried out using surface sea water from
19°08' N latitude, 105°29' W longitude
(BT Station 23-6). The 250-ml. reagent
bottles were filled completely with sea water
and NaHC1^, (k uc) was added carefully with
a syringe. Immediately after filling with
sea water, the bottles were sampled for their
bacterial counts by the pour-plate technique.
80 _
TABLE 8
COMPARISON OF RINSED AND AUTOCLAYED BOTTLES IN
TERMS OF BACTERIAL NUMBERS AND C1402 ASSIMILATION
Rinsed
bottles
Autoclaved bottles
Time, hours
Counts/minute
i Bacteria /ml
Counts/minute
Bacteria/ml
0
210
.
79
1
78
1,700
6l
700
2
78
3,500
122
900
1*
220
3,100
289
1,200
8
375
i3,itoo
I+78
22,000
18
622
7)+, 000
1,500
1*90,000
TABLE 9
COMPARISON OF RINSED AND AUTOCLAVED BOTTLES IN
TERMS OF BACTERIAL NUMBERS AND C1^ ASSIMILATION
Rinsed
bottles
Autoclaved
. bottles
Time, hours
Counts/minute
i Bacteria/ml
Count s/minute
Bacteria/ml
0
930
610
2
55
2,200
^5
1,300
1*
101
8,500
51*
3,300
6
111
79,000
122
28, 000
8
126
890, 000
ll+9
38,000
13
238
750,000
21*8
130,000
18
291
1,900,000
309
!*, 600, 000
TABLE 10
Ik
BACTERIAL DEVELOPMENT AND C UPTAKE IN DUPLICATE LIGHT BOTTLES CONTAINING
SURFACE SEA-WATER SAMPLES FROM 19°08' N LATITUDE, 105°29' W LONGITUDE
Time, hours
Bact'
sria/ml
Count
; /minute
Sample 1
Sample 2
Average
Sample 1
Sample 2
Average
0
3,200
3,600
3,1*00
.
.
_
2
6,600
6,700
6,600
I7I*
168
171
1*
7,800
9,600
8,700
311*
321*
319
8
62,000
1*0,000
51,000
559
676
6l8
16
680,000
890,000
780, 000
950
912
931
2U
91*0,000
1,000,000
970,000
1,118
752
935
37-5
6,1*00,000
6,800,000
6,600,000
1,33!*
1,1*21
1,378
- 81
Since there was a lag in time of 1.5 hours be-
tween the time the zero hour bacterial counts
were plated and the time all of the bottles were
inoculated with NaHC-^Oj and placed in the water
bath, this should be taken into account in
following the bacterial populations. The
illuminance in the water bath was l^-OO i lUO
foot -candles. After 2, k, 8, l6, 2k, and 37-5
hour 6 of incubation at 25 ± 1°C, two bottles
were removed and the contents were plated in
duplicate for bacteria, and filtered to det-
ermine the C^ uptake by the organisms over the
particular time span tested. The results for
the bacteria/ml and the C-^ assimilation show-
ing the values for the replicates are tabulat-
ed for the light bottles (Table 10) and the
dark bottles (Table 11). The average C1 up-
take was 171 counts/minute in the light during
the first two hours, with approximate doubling
after four hours and again after eight hours.
After eight hours, the C1^ assimilation in-
creased by a factor of 1.5X during the next
eight hours and 1.3X during the last 20 hours
of the experiment. Meanwhile, the bacteria
were in the lag phase of growth for the first
four hours, after which they entered logarith-
mic growth, tapering off somewhat after l6
hours. It is interesting to note that the
development of the bacteria in the dark and
the light was very similar.
lk
During the first four hours the C fixation
was considerably suppressed in the dark compared
to the light. After two hours, 37 counts/minute
were recorded which increased to 51 counts/min-
ute after four hours. At this point, however,
the C uptake more than doubled during the
next two time intervals (up to l6 hours).
After this time, the C^ fixation proceeded at
about the same rate in the dark and in the
light .
The replication of the bacterial counts and
Cl* fixation Was quite good with two excep-
tions. After 2k hours of C1^ uptake in the light,
the duplicate bottles did not agree. The higher
figure is more consistent with the other results.
There was considerable disagreement in the dupli-
cation of the 37 .5-hour count /minute in the dark
but the average figure appears reasonable
(Table 11).
DISCUSSION
The necessity for maintaining dark-bottle
controls during productivity measurements
by the C^ method becomes evident upon exam-
ination of the data in Tables 10 and 11.
When employing radioactive C^ as an index of
assimilation of CO2 and productivity, dark-
bottle controls have not always been consider-
ed important as a correction factor (steemann
Nielsen, 1952) . In these experiments, dark-
bottle C fixation became very significant
after eight hours of incubation (half as
much C1**- fixation in the dark as in the
light after eight hours). As shown from
Tables 10 and 11, the error (dark bottle
fixation/light bottle fixation) which would
be incurred, if the dark bottles were not con-
sidered, would be 21.6°/0 after two hours,
15.5% after four hours, 17.6% after
eight hours, 31.6% after 16 hours, W>.8%
after 2k hours, and 48.6% after 37-1/2 hours.
Thus, even during the customary incubation
period (up to eight hours), the error might
be expected to fall between 15 and 22°/, .
The effect of bacteria on the total C1^ fix-
ation i6 still somewhat uncertain. However,
the bacterial populations were very similar
in both the light and dark bottles and their
influence could be compensated for by using
dark-bottle controls. (The light source in
these experiments was artificial, not sun-
light, however.) This conclusion is support-
ed by the results of Vaccaro and Ryther (1951*-).
There was no indication in our studies of an
"antibiotic" effect wuch as that reported
by Steemann Nielsen (1955) for the fresh-water
green alga, Chlorella pyrenoidosa, and the
marine diatom, Thalassiosira nana.
Some calculations are presented for estima-
tion of the magnitude of bacterial C uptake
by the Wood-Werkman reaction. If one assumes
that an average marine bacterium is a short
rod (l micron long by 0.5 micron in diameter),
the volume of one bacterium would be 2.0 x
10-13 cc. At the end of 37.5 hours of in-
cubation, the volume of allrecorded bacteria
(6 x 109 cells/l) would be 1.2 x 10"3 cc/l.
If 80°/o of this volume is considered as
82
TABLE 11
lU
BACTERIAL DEVELOPMENT AND C UPTAKE IN DUPLICATE DARK BOTTLES CONTAINING
SURFACE SEA-WATER SAMPLES FROM 19°08' N LATITUDE, 105°29' W LONGITUDE
Time, hours
Bacteria/ml
Sample 1 Sample 2
Average
Count/minute
Sample 1 Sample 2
Average
0
2
k
8
16
2k
37.
3,200
5,600
8,500
58,000
660,000
1,600,000
5,200,000
3,6oo
5,100
7,900
59,000
390,000
1,700,000
5,700,000
3,^0
5, too
8,200
59,000
520,000
1,700,000
5,500,000
kk
59
139
3^5
te.5
872
30
^3
79
2 lj-2
1+51
H67
37
51
109
29U
14-38
670
TABLE 12
BACTERIAL FIXATION OF CARBON DIOXIDE CARBON IN THE DARK IN SURFACE
SEA WATER COLLECTED AT 19°08' N LATITUDE, 105°29' W LONGITUDE
Time, hours CO^/C in dark CO /C fixed by bacteria % of CO /C fixed by bacteria
Hg/1
Hg/1
"in the dark
2
0.18
k
0.25
8
0.52
16
l.fcl
2k
2.0
37-5
3.2
0.0055
0.0088
0.066
0.55
1.65
6.6
3
k
13
39
79
206
83 -
moisture content (Porter, I9I+6) then 2.1+ x
10" cc/l would be the dry volume or, multi-
plying by the average specific gravity, -
1.1 (Ruffilli, 1933), the dry weight of the
bacterial cells would equal 2. 61+ x 10"^ g/l.
If 50% of the dry weight of the bacterial
cells is considered as carbon (Porter, I9I+6) ,
then 1.32 x 10"^ g/l is the calculated weight
of total carbon. Since it has been estimated
that about 5°/ of cell carbon of heterotrophic
bacteria may be fixed by the Wood-Werkman re-
action, approximately 6.6 x 10"° g/l of C^
could have been fixed by the heterotrophic
bacteria at the end of the 37.5-hour period in
this experiment.
The total C02/C fixed in any of the sets of
bottles may be calculated as follows :
Total C02/C fixed/L =
Total COg/C present in mg/l x
count s/min/250 ml recovered x 1+ 4-
Count/min added x 1+
In this experiment 5,222,500 counts/minute (as
measured with our apparatus) of NaHC-^Oo were
added to each 250-ml. reagent bottle. The
total carbon dioxide carbon in the, surface sea-
water sample was approximately 25 mg/l.
Calculating the total carbon dioxide fixed in
each set of bottles in this manner and compar-
ing these values with the estimates of the
amount of C^ fixed by heterotrophic bacteria
for each period, an estimation of the percent-
age of COg/C fixed by the bacteria can be
obtained, as shown in Table 12 for each test
period in the dark.
Steemann Nielsen (1952) has estimated that
the amount of organically bound C^ is not a
completely accurate measure of the gross produc-
tion by photosynthesis since C-^02 is actually
assimilated at a rate 6°/0 slower than C-^Og.
In addition, Steemann Nielsen (1952) applies a
correction of 1+°/° of the photo synthetic in-
tensity at optimum light intensity in a four-
hour experiment for the loss of C through
the respiration of substances produced during
the experimental period. Thus, a 10°/o correc-
tion is applied. Steemann Nielsen neglects the
1^
negative correction due to C assimilation
in the dark which he estimates at l°/o, as
mentioned previously. However, in these
experiments the dark fixation of C^ was 15
to 20 times the dark fixation reported by
Steemann Nielsen (1952). The correction for
isotopic fractionation and respiration were
not applied in these calculations .
Various considerations which may affect the
calculations presented in Table 12 should be
mentioned. For example, the size of marine
bacteria is variable (ZoBell and Upham, I9I+I+) .
An increase in the length of the rod- shaped
cells from one micron to two micra would
double the importance of the bacteria in the
foregoing calculations. However, since marine
bacteria are generally veiy small, the values
used are considered reasonable. In addition,
the error in the pour-plate technique may be
considerable. It has been estimated that
only 1 to 10°/o of the bacteria present in a
sample are recorded by this method (ZoBell,
19^), which would increase their importance
in these calculations by at least a factor of
10. Little is known of the abundance or impor-
tance of chemosynthetic bacteria in the marine
environment which utilize carbon dioxide as
their sole source of carbon.
The various influences of bacteria, phytoplank-
ton, zooplankton and other components in the
marine ecosystem on Cl^ assimilation may
perhaps be elucidated by studies on pure
cultures and simple mixed populations. The
uptake of C^ by various members of the marine
population in pure cultures and in natural
mixtures should provide much additional
information on the actual uptake of C^ by
these organisms as well as offer more defini-
tive results concerning the effect of their
mutual interrelationships. These experiments
have been planned.
84 -
SUMMARY
1. Determinations of bacterial increases and
C-^Og fixation in surface samples of tropical
Pacific sea water contained in cleaned as well
as in cleaned and autoclaved 250-ml. reagent
bottles incubated in the light indicated that
both the bacterial populations and the C-^ up-
take were slightly lower in the autoclaved
bottles during the first few hours of incuba-
tion. The rate of increase of both the bac-
terial populations and the Cl^ uptake was
greater in the autoclaved bottles after the
first few hours and by the end of the l8-hour
incubation period their values were higher.
However, complete sterilization of the bottles
is not considered necessary for determinations
of productivity during the eight-hour period
generally employed, since the differences were
not great .
2. Dark-bottle fixation of C1^ varied be-
tween 15.5 and 21. 6% of light-bottle fixa-
tion during test periods up to eight hours
and up to almost 50*/o by 37-5 hours, indicat-
ing that such controls are necessary for
estimating productivity.
3. Bacterial C fixation was calculated as
varying between 3 and 13°/o of the total dark
fixation during the first eight hours of in-
cubation. The importance of bacteria would,
of course, be proportionally les6 in the
light.
h. The bacterial counts as determined by
the pour-plate technique were essentially the
same in both the light and dark bottles, and
there was no indication that either light or
antibiotics produced by the phytoplankton were
acting adversely on the bacterial populations.
BIBLIOGRAPHY
Gaarder, T., and H. H. Gran. 1927.
Investigation of the production of phytop-
lankton in the Oslo Fjord.
Rapp. Prov. Verb. Con6. Perm. Int. Explor.
Mer., Vol. k2, pp. 3-US.
Oppenhetmer, C. H., and C. E. ZoBell. 1952.
The growth and viability of sixty-three
species of marine bacteria as influenced
by hydrostatic pressure.
Jour. Mar. Res., Vol. 11, No. 1, pp.10-18.
Porter, J. R. 19^.
Bacterial Chemistry and Physiology.
J. Wiley and Sons, Inc., New York, pp.355.
Pratt, R., T. C. Daniels, J. J. Eiler,
J. B. Gunnison, W. D. Kumler, J. F. Oneto, and
L. A. Strait. 19^.
Chlorellin, and antibacterial substance
from Chlorella. Science, Vol. 99,
PP. 351-352.
Riley, G. A. 1938.
Plankton studies. I. A preliminary in-
vestigation of the plankton of the Tor-
tugas Region. Jour. Mar. Res., Vol. 1,
PP. 335-350.
Riley, G. A. 1939-
Plankton studies II. The western North
Atlantic, May-June, 1939.
Jour. Mar. Res., Vol. 2, pp. 1^5-l62.
Riley, G. A. 19^1a.
Plankton studies. III.
Bull. Bingham Oceanogr.
No. 3, PP. 1-93-
Long Island Sound.
Coll., Vol. 7,
Riley, G. A. 19^1b.
Plankton studies. IV. Georges Bank.
Bull. Bingham Oceanogr. Coll., Vol. 7,
No. k, pp. I-73.
Ruffilli, D. 1933.
Studies on the specific gravity of bac-
teria. Biochem. Zeit., Vol. 263, pp.63-74.
"1^ methods
Ryther, J. H., and R. F. Vaccaro. 195^f
A comparison of the oxygen and C-1
of measuring marine photosynthesis. Jour.
Cons. Int. Explor. Mer., Vol. 20, No. 1,
pp. 25-3^.
Ryther, J. H. 1956a.
Photosynthesis in the ocean as a function
of light intensity.
Limn, and Oceanogr., Vol. 1, No.l, pp.6l-70.
Ryther, J. H. 1956b.
The measurement of primary production.
Limnology and Oceanography, Vol. 1, No. 2,
pp. 72-8^.
85
Steemann. Nielsen, S. 1951. Wood, H. G., C. H. Werkman, A. Hemingway, and
Measurement of the production of organic A. 0. Hier. 19^1.
matter in the sea by means of carton-lV. Heavy carton as a tracer in heterotrophic
Nature [London), Vol. 167, p. 68V. carton dioxide assimilation.
Jour. Biol. Chem., Yol. 139, PP- 365-376.
Steemann Nielsen, E. 1952.
The use of radioactive carton (C.J for ZoBeli, C. E., andD. Q. Anderson. 1936.
measuring the organic production~of car- Otservations on the multiplication of
ton in the sea. tacteria in different volumes of stored
Jour. Cons. Int. Explor. Mer., Yol. 18, sea vater and the influence of oxygen
So. 2, pp. 117-lkO. tension and solid surfaces.
Biol. Bull., Yol. 71, pp. 32^, -3te-
Steemann Nielsen, E. 1951*-.
On organic production in the oceans. ZoBeli, C. E., and H. C. Upham. I9W+.
Jour. Cons. Int. Explor. Mer., Vol. 19, A list of marine tacteria including
No. 3, pp. 309-326. descriptions of sixty new species.
Bull. Scripps. Inst. Oceanogr., Yol. 5,
Steer-ann Mielsen, 2. 1955a. No. 2, pp. 239-292.
The production of antibiotics ty plankton
algae and its effect upon tacterial ZoBeli, C. E. 19^6.
activities In the sea. Marine Microtiology: A Monograph on
Marine Biology and Oceanography Suppl. Hydrotacteriology.
to Yol. 3 of Deep-Sea Reserch, pp. 281-286. Chronica Botanica Co., Waltham, Mass.
Steers." Nielsen, E. 1955b.
An effect of antitiotics produced ty plank-
ton algae.
Nature (London), Yol. 176, p. 553-
Utter, M. ?., and H. G. Wood. 1951.
Mechanism of fixation of carton dioxide ty
.-.eterotrophs and autotrophs. Advances in
Enzymoi., Vol. 12, pp. 1*1-151.
Yaccaro, R. ?., and J. H. Ryther. 195k.
The tactericidal effects of sunlight In
relation to "light" and "dark" tottle
photosynthesis experiments.
•Jour. Cor.s . Int. Explor. Mes., Yol. 20,
No. 1, pp. 18-2^
Wood E. G., and C. H. Workman. 1938.
The utilization of C02 ty the propionic
acid tacteria.
Biochen. Jour., Yol. 32, pp. 1262-1271.
Wood, H. G., and C. H. Werkman. 19^0.
The relationship of tacterial utilization
of CO2 to succinic acid formation.
Biochem. Jour., Yol. 3S pp. 129-138.
to -
TEE EFFECTS OF ORGANIC ARD INORGANIC KlCRONUTRISNTS OR THE ASSIMILATIOR OF C
BY PLARKTORIC COMKURTTISS ARD OR BACTERIAL MULTIPLICATION IR TROPICAL PACIFIC
SEA WATER
Galen E. Jones and William H. Thoma6
lh
Predatory concepts of life in the sea vere
emphasized throughout the early history of
marine biology. However, various line of
evidence prompted Lucas (l9^7> 19^> 1955) to
propose that microorganisms in the sea can
interact in a nonpredatory manner by means of
external metabolites . In this vay certain
organisms might influence the activities of
others by producing essential nutrients or by
removing or excreting inhibitory substances.
Certain marine phytoplankton require growth
factors such as thiamin, cyanocobalamin, and
biotin (Provasoli and Pintner, 1953; Lewin,
1951*-; Sweeney, 195^j Droop, 1957; and Johnston,
1955) • Requirements for amino acids, purines,
pyrimi dines, and other grovth factors have
also been shown for some marine bacteria
(Ostroff and Henry, 1939; MacLeod et al., 1951*-;
Jones, 1957) • Some marine algae contain a
great diversity of grovth factors (Ericson,
1953a and b; Ericson and Carlson, 1953) — -1
could presumably supply such factors to other
marine organisms. The literature on the exist-
ence of such factors in sea vater has been re-
viewed by Vallentyne (1957)-
The present paper reports experiments per-
formed at sea on the effects of small con-
centrations of added organic substances on bac-
terial grovth and C^-^02 assimilation by organ-
isms in pelagic sea-vater samples. The effects
of additions of certain inorganic substances on
these processes were also studied.
M3TH0DS
,1^
The radioactive C method for measuring
organic productivity (Steemann Nielsen, 1951,
1952) was utilized to determine the amount of
C assimilation by microorganisms in light
and dark bottles containing pelagic surface sea
vater vhen microquantities of organic and in-
organic nutrient pools vere added. Reagent
bottles of a 250-ml. capacity vere thoroughly
cleaned vith detergent, rinsed vith 10% HC1,
and finally vith sea vater (five or six times
immediately before use). All of the samples
vere surface sea vater collected in a clean
plastic bucket. After a 220-ml. sample of
sea vater vas added to the bottles, the
number of marine heterotrophic bacteria in
the sample vas determined using the pour-
plate method and peptone-yeast extract agar
(Oppenheimer and ZoBeli, 1952) . Similar en-
umerations vere made at the end of the period
of incubation vith C1^. Plates vere poured
on a suspended table vhich vas adequate to
compensate for the roll of the ship in the
moderate seas experienced on this cruise.
The bacterial plates vere incubated for three
days in the dark at 30 t 1*C before ex-
amining for bacterial numbers vith a Quebec
colony counter. Appropriate dilutions vere
made vith sterile vater blanks vhen high counts
vere anticipated. Uninoculated control plates
vere maintained in all cases to check the
sterility of the medium, the sterility of the
disposable plastic petri dishes employed and
the incidence of contamination due to handl-
ing aboard ship. These uninoculated plates
vere sterile in most cases.
Organic constituents vere added as pools to
certain sea-vater samples to give the
following final concentrations:
Vitamin pool 1 (Y-l) ug/100 ml of sample
folic acid 1.0
thiamin chloride 1; . ;
riboflavin 5.0
pyridoxine hydrochloride 5.0
calcium pantothenate =:._
nicotinamide =:.:
choline hydrochloride 100.0
inositol 100.0
para-aminobenzoic acid 5.0
biotin 0.05
-
Vitamin pool 2(v-2)
pyridoxal phosphate
pyridoxamine dihydrochloride
cyanocobalamin
vitamin A
acetylcholine chloride
ascorbic acid
carotene
nicotinic acid
Vitamin pool 3 (V-3)
calciferol
tocopherol
rutin
menadione
ug/lOO ml of sample
5.0
5.0
0.015
10.0
10.0
10.0
10.0
50.0
ug/100 ml. of sample
10.0
10.0
10.0
10.0
The PI metal stock solution (Provasoli et al,
1957) furnished micronutrients and EDTA in
the following final concentrations:
Constituent mg/lOO ml of sample
These vitamin pools vere prepared in 50°/o
ethyl alcohol and added in 0.1-ml. amounts
per 100 ml of sample sea water. Pools of
purine and pyrimidines (pp) (adenine,
adenosine, adenylic acid, guanine, guanosine,
uracil, cytidylic acid thymine, xanthine, and
hypoxanthine ) , essential amino acids (EAA)
(valine, isoleucine, leucine, threonine, phen-
ylalanine, trytophane, lysine, arginine, his-
tidine, and methionine), and nonessential amino
acids (NKAA) (glutamic acid, aspartic acid,
serine, proline, cystine, glycine, alanine, and
tyrosine) were made up in double distilled water
in one-mg. amounts per 100 ml. of sample (final
concentration) . The purine and pyrimidine pool
as well as the nonessential amino acid pool was
sterilized in the autoclave, whereas the
essential amino acid pool was passed through an
ultrafine Morton s inter ed-glass filter.
Organic complexes were added to some samples
to give the following final concentrations:
soil extract (from Scripps garden soil)
(Sweeney, 1951) 1 ml/100 ml of sample; yeast
extract (Difco) 0.001 g/100 ml.
Inorganic substances were added to give the
following final concentrations:
KHO3...IO ugm at N0o/L of sample j KgHP01f...l ugm
at POl^/L of sample} PI Metals... 3 ml/100 ml of
sample .
Na2EDTA
Fe
B
Mn
Zn
Cu
Co
3-0
0.03
0.6
0.12
0.015
0.00012
0.0003
The 250-ml. glass-stoppered reagent bottles
were incubated in a water bath which was
illuminated (fluorescent lighting from a
battery of long bulbs at an illuminance of
1250 1 150 foot-candles) through a glass
bottom. The temperature of the water bath
was maintained as close to the temperature of
the surface sea water as possible. Black
bottles (prepared by careful covering with
black masking tape) were incubated with the
light bottles as controls to distinguish
dark uptake of C1^ from photosynthetic fixa-
tion. After the bottles were incubated in
the presence of added nutrients for approxi-
mately four hours, the NaHC-'- 0o was added
to the water samples from a sterile ampule
and the samples were incubated for two to
four additional hours.
At the end of the incubation period, equal
volumes of water (2.0 ml) were removed from
each sample bottle after shaking and plated
in duplicate, as described above, to determine
the numbers of heterotrophic marine bacteria.
In some experiments the zero-hour count was
subtracted from the final bacterial popula-
tion to give the bacterial increase.
The water remaining in each bottle was filter-
ed through an BA Millipore filter (0.1*5 t
0.02 microns, Millipore Filter Corporation,
Watertown, Mass . ) which retained all of the
larger particulate matter; the filter wa6
then washed with more than 100 ml of sea water
and dried in a desiccator for at least 21*-
hours over silica gel. The radioactivity on
the filter pad was measured in a proportional
flow counter (Nuclear Measurement Corporation,
PC-1).
RESULTS
nlkr
In the first experiment, the C O2 assimila-
tion and bacterial numbers in surface sea
water from a poorly productive area (30*01' N
latitude, ll6%9' W longitude) west of Baja
California were determined in both light and
dark bottles. The samples were incubated for
seven hours in the illuminated water bath at
l8 + 1*C. The results are shown in Table 13 .
The C-^Og assimilation in the light was 1.6
times that in the dark and the bacterial in-
crease was almost four times that in the dark.
In this experiment the photosynthesetic activ-
ity of the phytoplankton apparently stimulat-
ed the bacterial population, presumably be-
cause of the metabolic by-products of the marine
algae. The importance of the dark-bottle con-
trols was emphasized by the 63°/» dark fixation
in this experiment. In the rest of the experi-
ments dark-bottle controls were utilized wherev-
er possible.
In the next experiment the effect of inorganic
nutrients on C^Oo fixation and bacterial
populations was determined in poorly productive
water just north of the Alijos Rocks west of
Baja California (26*50* N latitude, 116*13' W
longitude; Station BT - 0 - 7) . NO3, POI4., and
PI metals were added to one pair of bottles .
Pairs of these inorganic additives were added
to other sets of bottles containing the sea-
water sample . Soil extract was added in an-
other set of bottles to the NOo, POj^, and PI
metals as a final treatment. Light- and dark-
bottle controls containing no additions to the
sea-water sample were incubated at 20 to 2^*C
for a total of six hours with the treated
samples. After four hours 5,222,500 counts/
minute of NaHC1 0o were added to each bottle
from sterile ampules. The results of thi6
experiment appear in Table Ik.
Little or no effect on C1 0g assimilation
due to the addition of the inorganic elements
was observed in this water mass. However,
the populations were permitted to adjust to
the conditions in each set of bottles for only
four hours before adding the NaHC-^Oj, so that
the period allowed for the uptake of C1^ may
not have been long enough to accentuate the
differences in fixation in the different treat-
ments, particularly in a poorly productive area
such as that under study.
The results in Table lk regarding the
bacteria are not particularly informative.
PI metals and POi,. appear to be the most
stimulatory to bacterial development in this
water, but there is no way to determine from
these results whether the increase in bacteri-
al populations is due to by-products from the
phytoplankton or to direct stimulation from
the inorganic substances. In thi6 case, the
large number of bacteria in the dark compared
with the light supports the results of
Steemann Nielsen ( 1955a and b).
The organic pools were compared with the in-
organic additions in the following experiment.
A surface sea-water sample from a poorly pro-
ductive area south of Alijos Rocks off Baja
California (21°3V N latitude, 110* 1*9 ' W
longitude; BT - 1 - 5) was treated in duplicate
with the following nutrient additions: vitamin
pool 1, 2, and 3> purine and pyrimidine pool;
KNO3; KgHPO^; and PI metals. The surface sea-
water temperature was 28.1+°C. A combination
of all of the nutrient additions mentioned
above was added to one set of bottles, and
all of the vitamin pools, the purines and
pyrimidines, and the inorganic additions were
subsequently removed from the combined pool,
one at a time, in different experimental
samples. Finally, another treatment contained
KN0., KgHPOi,. and 0.025 ml of Hoagland-Arnon's
(1950) trace-element solution. Twelve of the
bottles were selected at random and tested
for the initial bacterial numbers in the
bottles. To each bottle 5,222,500 counts/min-
ute of NaHC1^^ were added after four hours
of incubation at 30 _ 1*C and the bottles in-
cubated for an additional three hours. The
bottles were harvested in the usual way and
the bacterial counts determined. The results
are presented in Table 15.
The vitamin pools exerted an inhibitory effect
on the C 02 assimilation of the phytoplankton
in the light. Where the vitamin pools were
deleted from the bottles and the purine and
pyrimidine pool and the inorganic additions
(KNO3, KgHPO^ and PI metals) were added to the
water sample, the C1^ fixation was 1.5 times
a6 great as the untreated control during the
seven hours incubation. Wherever the vitamins
were present C-^Og uptake was depressed.
89
TABLE 13
Ik
C ASSIMILATION AND BACTERIAL INCREASE IN LIGHT AND DARK BOTTLES COLLECTED AT
30o01' N LATITUDE, ll6°^9' W LONGITUDE AFTER 7 HOURS INCUBATION AT l8 + 1°C.
Treatment
Bacterial increase/ml *
Counts/minute
Light bottles 1,700
Dark bottles hy
Increase due to light 1,2V?
* Average bacterial count at zero hour was 250 bacteria/ml,
619
389
230
M
TABLE 1^
0 ASSIMILATION AND BACTERIAL NUMBERS AFTER 7 HOURS INCUBATION AT
20 - 24°C IN THE PRESENCE OF INORGANIC ADDITIONS IN WATER COLLECTED AT
26°50' N LATITUDE, ll6°13' W LONGITUDE: STATION BT - 0 - 7-
Treatment
No additions - light
No additions - dark
N0„ PO, , PI metals - light
P0\ . - light
Bacteria/ml *
Counts/minute
3,
POr, PI metals - light
NO , PI metals - light
NO, PO, , PI metals, soil extract - light
* Average bacterial count (14 bottles,
2,060
k,oyo
3,960
2,900
5,000
3,320
3A50
at zero hour was 260 bacteria/ml
21+0
^5
276
209
260
228
21*0
90 -
TABLE 15
EFFECTS OF SMALL CONCENTRATIONS OF INORGANIC AND ORGANIC ADDITIONS ON BACTERIAL
MULTIPLICATION AND C1^ ASSIMILATION IN WATER COLLECTED AT 21° 3k' N LATITUDE,
110°^9' W LONGITUDE; STATION BT - 1 -5, INCUBATED 7 HOURS AT 30 - 1°C.
Treatment Light Bottles Dark Bottles
Bacterial increase/ml* c/min Bact.. increase/ml* c/min
No additions
Vitamin pools 1, 2, 3
Purine and pyrimidine pool
Inorganic pool
Purine and pyrimidine pool
Inorganic pool
Vitamin pools 1, 2, 3
Inorganic pool
Vitamin pools 1, 2, 3
Purine and pyrimidine pool
KNO,, K2HP0^, H and A trace
elements
* Average number of bacteria/ml before incubation was 2^+00 - 600.
5,250
211
K
.lto
97
29,61+0
127
2k,
,350
102
15,660
308
23;
,oi<o
109
12,5^
ltl
10;
,200
158
30,5to
128
-
130
11,220
153
_
_
- 91
The purine and pyrimidine pool had the most
marked stimulatory effect on the bacterial
multiplication in the various treatments.
When the purine and pyrimidine pool was re-
moved from the complete complement of addi-
tives, the bacterial increase in both the
light and dark bottles were little more than
twice the number of those in the untreated
controls. Increases of sixfold were recorded
(Table 15) when the purine and pyrimidine
pool was present. The bacteria did not appear
to be inhibited by the vitamin pools. In fact,
the vitamins stimulated bacterial development
somewhat in all cases. The inorganic addi-
tions failed to promote more than a twofold
increase in the number of bacteria.
In the experiment reported in Table 15, C1^
assimilated by the phytoplankton was inhibited
by the vitamin pools. Since these vitamin
pools were prepared in 50° /o ethanol to pre-
serve their sterility, the possibility that
the alcohol per se was depressing the C^ up-
take by the plants was considered. The final
concentration of ethanol was 0.15/° •
In the following experiment, the ethanol was
removed from the vitamin pools by hot-air
evaporation at 35°C. After the alcohol was
removed, the original volume of the pools
was reconstituted by adding distilled water.
A surface sea-water sample was collected in
the same manner as before at ll4-°27' N latitude,
98°58' w longitude} (Station BT - 5 - 7) • The
temperature of the surface sea water was 28.0°C.
Control bottles to which no additions were made
were prepared in three different ways: 1) cleans-
ed and rinsed with 95°/o ethanol, followed by
five or six rinses with the sample sea water,
2) cleaned with a detergent, rinsed with sea
water, rinsed with 10°/. HC1, rinsed five or six
more times with sea water and autoclaved for
15 minutes at 15 lbs. pressure, 3) bottles rins-
ed five or six times with sample 6ea water. All
of the bottles to which nutrients were added
were prepared as described in the first method
with 95*/o ethanol followed by five or six
rinsings with sample sea water.
The same additions were made in this experi-
ment as in the last experiment (Table 15) ex-
cept that the final inorganic treatment (KNOo,
K/>HP0l, and Hoagland and Arnon's trace ele-
ments) were not repeated. These bottles were
incubated in the illuminated water bath for
four hours at 28 ± 20C. After four hours
both the light and dark bottles were removed
and 5,222,500 counts/minute NaHC11^ were
added. The bottles were returned to the water
bath for two more hours of incubation (total
of six hours), after which the final bacterial-
assay plates were poured and the contents of
the bottles filtered for C1^ uptake. The re-
sults are presented in Table l6. The initial
bacterial numbers, which varied between 6I4O
and 1,250 bacteria/ml, were subtracted from
the final count to determine the bacterial
increase/ml.
These results confirm the conclusion of the
previous experiment (Table 15) regarding the
inhibition of photosynthesis by the vitamin
pools . The removal of the ethanol from the
vitamin pools made little difference on the
C1^02 assimilation of the phytoplankton in
the presence of these pools. Consequently,
it may be concluded that the vitamin pools
themselves exerted some inhibitory effect on
C1* uptake by phytoplankton in the light.
Where the vitamin pools were omitted and the
purine and pyrimidine pool as well as the in-
organic additions were added, photosynthesis
was almost doubled compared to the untreated
control (Table l6) .
The dark fixation in the treated bottles was
1.5 to 2.0 times greater than that in the
untreated control. The greatest dark fixa-
tion was in the sample containing the purine
and pryimidine pool plus the inorganic addi-
tions as in the light. The bacterial increase
in this treatment was considerable, 1+1,000
bacteria/ml. as compared with 12,1)00 bacteria/
ml. in the control dark bottles.
The most marked bacterial increase in this ex-
periment was that resulting from the combined
treatment (vitamin pools, purine and pyrimidine
pool, and inorganic additions), 82,000 bac-
teria/ml in the light and 28,000 bacterial/ml
92 -
TABLE 16
EFFECTS OF SMALL CONCENTRATIONS OF INORGANIC AND ORGANIC ADDITIONS ON BACTERIAL
MULTIPLICATION AND C14 ASSIMILATION IN WATER COLLECTED AT l4°27' N LATITUDE,
96058' W LONGITUDE (BT - 5 - 7) AND INCUBATED FOR 6 HOURS AT 28 +. 2° C.
Treatment
Light Bottles
Bact . increase/ml
c/m Dark Bottles
Bact . increase/ml c/m
No additions (alcohol rinsed)
No additions (autoclaved)
No additions (rinsed only-
Vitamin pools 1, 2, 3
Purine and pyrimidine pool
Inorganic pool
Purine and pyrimidine pool
Inorganic pool
Vitamin pools 1, 2, 3
Inorganic pool
Vitamin pools 1,2, 3
Purine and pyrimidine pool
20,500
569
13,300
65^
21,000
6l0
82,000
152
26,600
976
2l+,000
121
5^,000
119
12,ll00
91
28,000 ikk
^1,000 208
31^00 1^3
65,000 167
93
TABLE IT
lk ,
C FIXATION AND BACTERIAL INCREASES/ml IN A SURFACE SEA-WATER SAMPLE COLLECTED AT
09o0lj-' N LATITUDE, 89°13' W LONGITUDE AFTER 8 HOURS INCUBATION AT 25 - 1°C IN THE
LIGHT.
Treatment
Bacterial increase/ml *
Counts/minute
No additions
59,000
Combination of all
1,960,000
Vitamin pool 1 (V-l)
55,500
Vitamin pool 2 (V-2)
59,500
Vitamin pool 3 (V-3)
6ij-,500
Purine and pyrimidine
(PP)
76I+, 500
Essential amino acids
(EAA)
256,000
Non-essential amino acids (NEAA)
182,000
Yeast extract
2,165,000
Soil extract
339,000
Tween 80
33^,000
273
392
308
261
162
335
I4.25
Uoo
281
779
* Average bacterial count /ml at zero hour was ^,920.
TABLE 18
C ASSIMILATION AND BACTERIAL INCREASES/ml IN LIGHT AND DARK BOTTLES TREATED
WITH VARIOUS ORGANIC GROWTH FACTORS IN SURFACE SEA WATER COLLECTED AT 09° 38'
N LATITUDE, 89°35' W LONGITUDE, AFTER 8 HOURS OF INCUBATION IN 25 + 1° C .
Treatment
Light Bottles
Bact. increase/ml
c/m
*
Dark Bottles
Bact. increase/ml*
c/m
No additions
18, 000
26^
32,000
97
Biotin
26,000
253
26,000
81
Thiamin
26,000
288
2l+,000
76
Cyanocobalamin
28,000
35^
18,000
113
Methionine
17,000
280
9,000
in
Cystine
15,000
135
16,000
70
Tween 80
te,ooo
221
65,000
206
*Average bacterial count at zero hour was 8,670 bacteria/ml.
9k -
in the dark. Thus, while the presence of
vitamins depressed C^-^Og uptake by the phyto-
plankton in the light, bacterial multiplica-
tion was enhanced. This may he due to the ac-
tion of the vitamins on the bacteria directly
or to the release of organic substances from
the phytoplankton.
The various methods for preparing the bottles
in the otherwise untreated samples had no
appreciable affect on the C^-^j assimilation
by light -incubated phytoplankton in the
samples, although the autoclaved bottles show-
ed a slightly increased C^^C^ uptake and about
one-third fewer bacteria.
An experiment of the same type was prepared to
determine the effect of the individual pools
and other complexes on C^^Do fixation and bac-
terial populations. Surface sea water was
collected at 09*0k' N latitude, 89°13' W
longitude (Station BT - 9 - 2k), dispensed in
250-ml glass stoppered reagent bottles, and
treated as follows: no treatment (control),
combination of all that follows, vitamin pool
1 (V-l), vitamin pool 2 (V-2), vitamin pool 3
(V-3)j purines and pyrimidines (pp), essential
amino acids (EAA), nonessential amino acids
(NEAA), yeast extract (0.001*/o), soil extract
(1.0°/.), and Tween 80 (0.001°/.). The temp-
erature of the surface sea water was 2k-.~j'C.
The bottles were incubated at 25 1 1° C in the
illuminated water bath for four hours before
1,21*6,300 counts/minute of NaHC1^ were added.
The bottles were returned to the water bath
and incubated for a total of eight hours.
Owing to lack of space in the water bath, dark
bottles were not included in this experiment.
The results of the bacterial increases per ml
and the Cl^s assimilation in the light appear
in Table 17 .
None of the vitamin pools stimulated either
{jl* uptake or bacterial development to any
extent. Vitamin pool 3 (Y-3) decreased the
C^ fixation by more than 1/3 of the untreated
control. The surface active agent, Tween 80,
( a complex mixture of polyoxyethylene ethers
of mixed partial oleic ethers of sorbitol anhy-
drides) increased C1^ fixation by a factor of
three over the untreated sample, the most
pronounced stimulation of C^ assimilation in
any of the treatments. The bacterial increase
was also appreciable for this treatment
(Table 17) .
The amino acid pools stimulated C^^* fixa-
tion in the light by a factor of about 1.5,
and the essential amino acids promoted a
slightly greater increase. These amino acid
pools also stimulated bacterial development;
the essential amino acids proved about 1.3
times as effective as the nonessential amino
acid pool (Table 17).
The purine and pyrimidine pool enhanced ClV>2
fixation only slightly hut stimulated the
bacterial numbers by a factor of thirteen
compared to the untreated controls . These re-
sults appear consistent with those of previous
experiments.
ill
The soil extract had no effect on C uptake
but increased the bacterial numbers by seven-
fold. The treatment containing yeast extract
was so cloudy and turbid by the end of the
eight-hour incubation period that, it could not
be filtered. Consequently, no C O2 fixation
data exist for this treatment. The bacteria
increased by a factor of 36 compared to the
untreated control which was by far the great-
est increase in the bacterial population ex-
cept where yeast extract was present in the
combined treatment. Although the combined
pools showed considerable stimulation compared
to the untreated controls, the values for
C^-^Oo fixation and bacterial increases were
lower than some of the individual treatments
(Table 17 ) . This effect was attributed to
the inhibitory properties of the vitamin pools.
Finally, an experiment was conducted to deter-
mine the effects of selected individual
substances from the pools on C^ O2 fixation
and bacterial development. Surface sea water
at 25.7°C was collected at 09*38' N latitude,
89*35 ' W longitude (Station BT - 9 - 36) and
dispensed in the 250-ml glass-stoppered rea-
gent bottles as in the other experiment. The
following additions were made to paired bottles
- 95
in the same concentrations as in the pools:
no additions, biotin (0.05ug/l00 ml), thiamin
(10.0 ug/lOO ml), cyanocobalimin
(0.015 ug/lOO ml), methionine (l mg/100 ml),
cystine (l mg/lOO ml), and Tween 80
(l mg/lOO ml). Bacterial plates were poured
as usual with the yeast extract -peptone agar
both before and after the eight -hour
incubation period. After ^.5 hours of
incubation at 25 ± 1°C in the illuminated
rater bath, 5,222,500 counts/minute of NaHC^Oo
tfere added to each bottle.
rhe results of this experiment, using both
Light and dark bottles, are presented in
rable 18.
Little can be concluded from this experiment.
Che most stimulatory addition for C^-^02
fixation in the light was 0.15 mug/ml of
:yanocobalamin (vitamin B]^) which increased
:1^02 uptake by a factor of 1.3 compared to
the untreated control in the light. The only
3ther significant difference from the untreated
:ontrol was where 10 ug/ml of the sulfur-
:ontaining amino acid, cystine, was added,
rtiich depressed the Cl1^ fixation by 4-9 c/o in
:he light.
Phe addition of 10 ug/ml Tween 80 doubled the
lark fixation. Methionine inhibited the dark
lptake of C1* by 58%.
l?he bacterial numbers were not influenced by
the organic growth factors to any appreciable
axtent, but the addition of Tween 80 increased
the bacterial population by a factor of 2.5
is compared with the untreated control in the
Light. Most of these supplements stimulated
aacterial development in the light as com-
pared to the untreated control. The bacterial
levelopment in the dark was greatest in the
antreated control and in the presence of
[Veen 80.
DISCUSSION AND CONCLUSIONS
In the series of experiments presented, an
attempt was made to determine whether certain
Drganic substances (NO3, P]0^, and trace
slements) and certain organic pools (vitamin,
amino acids, purines and pyrimidines, Tween 80,
yeast extract, soil extract, etc.) would
"trigger" increases in the C-^Og assimilation
processes of the phytoplankton or in the
bacterial populations in tropical Pacific sea
water. These experiments were carried out
immediately after water samples were collected.
Vitamin pools in the concentrations employed
did not stimulate C1^ fixation by the
phytoplankton samples from the tropical Pacific
Ocean. In most cases the Cl^02 uptake was
considerably inhibited by the vitamin pools in
the light (Tables 15-17). It is interesting
to note in Table 17 where the vitamin pools were
tested separately for their effect on the uptake
of Cl^ by the phytoplankton, that vitamin pool
1 was slightly stimulatory compared to the
untreated control, vitamin pool 2 exerted little
influence, and vitamin pool 3 was inhibitory.
One of the constituents of vitamin pool 3 is
menadione. Dam (19^) has demonstrated that
this vitamin is inhibitory to photosynthesis
in Chlorella due to direct toxic action on the
cells. This fact coupled with the observation
that various individual vitamins such as thiamin
and cyanocobalamin actually stimulated CIW2
fixation by the phytoplankton (Table 18) to a
small extent, suggests that the vitamins as a
group are not inhibitory to C^-^C^ fixation in
the light but that various inhibitory constitu-
ents of the vitamin pools may mask the effects
of other members of the pools. Vitamin pools
were not inhibitory to bacterial development
in any case. However, where the individual
vitamin pools were added to separate bottles
(Table 17), there was almost no stimulation
from any of the pools as compared to the
untreated control. In other experiments there
was some indirect information suggesting that
vitamins were stimulatory to bacterial
development (Tables 15 and 16) . Generally, the
vitamin pools were not as stimulatory as the
other organic pools tested (amino acids,
purines and pyrimidines) .
The purine and pyrimidine pool enhanced the
development of marine bacteria and increased
the Cl^02 uptake of the phytoplankton (Tables 15-
17) . Purines and pyrimidines in natural waters
have received little attention from previous
investigators, but some evidence exists
suggesting the limited distribution of the
pyrimidine, uracil, and an unidentified purine
in pelagic sea water (Vallentyne, 1957;
- 96
Belser, 1957) • The significance Of these com-
pounds in the ecology of the sea is strongly
implied by the experiments presented in this
paper .
Amino-acid pools stimulated "both C^Og
uptake by the phytoplankton and bacterial
development (Table 17) • Some marine bacteria
have been shown to require certain amino acids
(Ostroff and Henry, 1939; MacLeod et al., 195^;
Jones, 1957). In addition, Fogg (1952J
reported that the blue-green alga, Anabaena
cylindrica, produced equal amounts of
extracellular and intracellular polypeptide
nitrogen. It is highly probable that
proteinaceous compounds in the sea exert
considerable influence on the mutual
interrelationships between marine phyto-
plankton and bacteria.
The great increase in C^-^t^ assimilation by
the phytoplankton in the presence of 0.001°/o
of the surface active agent, Tween 80,
(Table 17) which was not confirmed by a later
experiment (Table l8) will require further
consideration. It is of interest to note
that the bacterial numbers were increased
markedly in both experiments in the presence
of the Tween 80. Inorganic additions did
not appear to enhance C^02 fixation or
bacterial development appreciably. This may
be due to a lack of organic growth factors in
tropical Pacific sea water rather than to a
limitation of inorganic nutrients.
The exposure of the phytoplankton in the sea-
water samples to the NaHCl^K^ for short
periods of time (2 to h hours) may not have
been sufficient to allow appreciable
differences in Cl^C>2 assimilation to take
place in all cases. If the cells were
deficient in one or more of these nutrients,
it might take some time for uptake to be
reflected by the photosynthetic mechanism.
For example, it takes about 2h hours for
Scenedesmus cells to recover from nitrogen
deficiency to the extent of containing the
amount of protein found in normal cells
(Thomas and Krauss, 195*0 • However, in the
present work short experimental periods were
chosen so that photosynthesis could be
measured without measuring phytoplankton
growth. In subsequent experiments of this
type the times could be varied.
The importance of dark-bottle controls for all
treatments in experiments of this type is
evident from an examination of any of the values
obtained for dark fixation compared with light
fixation. This conclusion is supported by that
of Jones et al. (this volume).
These experiments provide preliminary informa-
tion from the natural environment which can be
used for future detailed culture and
photosynthetic experiments in the laboratory.
For instance, the development of culture media
for pelagic phytoplankton and marine bacteria
might be facilitated by the inclusion of some
of these substances, especially purines,
pyrimidines and amino acids, in the media.
Specific effects of these substances on the
photosynthetic mechanisms of the phytoplankton
and requirements by the bacteria may be
determined in pure culture .
ACKNOWLEDGEMENTS
The authors would like to express their sincere
appreciation to Dr. William L. Belser, Scripps
Institution of Oceanography, for his help in
formulating the organic constituents tested and
for critically reviewing the manuscript. In
addition, the authors would like to thank
Mr. Donald W. Lear and Mr. Harold L. Scotten,
Scripps Institution of Oceanography, for their
help in preparing for the cruise.
BIBLIOGRAPHY
Belser, W. L. 1957.
The use of auxotrophic mutants of a marine
bacterium for the bioassay of organic
micronutrients in the sea. Bacteriol.
Proc, pp. 30.
Dam, H. I9M4.
Vitamin K in unicellular photosynthesizing
organisms. Amer. Jour. Bot., Vol. 31,
PP. l4-92-i4-93-
Droop, M. R. 1957.
Auxotrophy and organic compounds in the
nutrition of marine phytoplankton. Jour.
Gen. Microbiol., Vol. 16, pp. 286-293.
97 -
Ericson, L. E. 1953a.
On the vitamin B12-, folic acid-, and
folinic acid groups of factors, and on
the occurrence of these vitamins and
of niacin, pantothenic acid and amino
acids in a number of marine algae.
Thesis, Uppsala University, Sweden,
PP. 1-79.
Ericson, L. E. 1953b.
Further studies on growth factors for
Streptococcus faecalis and Leuconostoc
citrovorum in marine algae. Arkiv for
Kemi, Vol. 6, No. 8, pp. 503-510.
Ericson, L. E., and Blenda Carlson. 1953-
Studies on the occurrence of amino
acids, niacin and pantothenic acid in
marine algae. Arkiv for Kemi, Vol. 6,
No. 1+9, PP- 511-522.
Fogg, G. E. 1952.
The production of extracellular
nitrogenous substances by a blue-green
alga. Proc. Roy. Soc, B, Vol. 139,
PP. 372-397.
Hoagland, D. R., and D. I. Arnon. 1950.
The water culture method of growing plants
without soil. Calif. Agr.Expt .Sta.Circ. ,
Vol. 3V7, Revised edition.
Johnston, R. 1955.
Biologically active compounds in the sea.
Jour. Mar. Biol. Assoc. U.K., Vol. 34,
pp. 185-195.
Jones, G. E. 1957-
The effects of organic metabolites on
the development of marine bacteria.
Bacterid. Proc, pp. 16.
Jones, G. E., W. H. Thomas, and F. T. Haxo.
Preliminary studies of bacterial
growth in relation to dark and light
fixation of Cll402 during productivity
determinations, (this volume).
Lewin, R. A. 1951*--
A marine Stichococcus sp. which requires
Vitamin B]_2 (cobalamin). Jour. Gen.
Microbiol., Vol. 10, pp. 93-96.
Lucas, C. E. 19^7.
The ecological effects of external
metabolites. Biol. Rev., Vol.22, pp. 270-295.
Lucas, C. E. I9I4-9 .
External metabolites and ecological
adaptation. Symp.Soc .Expt .Biol., Vol. 3,
PP. 336-356.
Lucas, C. E. 1955.
External metabolites in the sea. Mar.
Biol, and Oceanogr. Suppl. to Vol. 3 of
Deep-Sea Res., pp. 139-148.
MacLeod, R. A., E. Onofrey, and M. E. Norris. 1954
Nutrition and metabolism of marine bacteria.
I. Survey of nutritional requirements.
Jour .Bacterid., Vol. 68, pp.680-686.
Oppenheimer, C. H., and C. E. Zobell. 1952.
The growth and viability of sixty-three
species of marine bacteria as influenced
by hydrostatic pressure. Jour .Mar .Res . ,
Vol.1, No.l, pp. 10-18.
Ostroff, Rose, and B. S. Henry. 1939.
The utilization of various nitrogen
compounds by marine bacteria. Jour .Cellular
Comp. Physiol., Vol.13, pp. 353-371-
Provasoli, L., and Irma J. Pintner. 1953.
Ecological implications of in vitro
nutritional requirements of algal flagel-
lates. Ann. N.Y. Acad.Sci. , Vol. 56, No. 5,
pp. 839-851.
Provasoli, L., J. J. A. McLaughlin and
M. R. Droop.' 1957.
The development of artificial media for
marine algae. Archiv. fur Microbiol.,
Vol.25, pp. 392-428.
Steemann Nielsen, E. 1951.
Measurement of the production of organic
matter in the sea by means of carbon-l4.
Nature (London), Vol.167, p. 684.
Steemann Nielsen, E. 1952.
The use of radioactive carbon (C1^) for
measuring the organic production of carbon
in the sea. Jour .Cons. Int. Explor.Mer. ,
Vol.18, No. 2, pp. 117-140.
- 98
Steemann Nielsen, E., 1955a.
The production of antibiotics by-
plankton algae and its effect upon
bacterial activities in the sea.
Mar .Biol, and Oceanogr .Suppl. to
Vol. 3 of Deep-Sea Res., pp.28l-286.
Steemann Nielsen, E., 1955b.
An effect of antibiotics produced by-
plankton algae. Nature (London),
Vol. 176, p. 553-
Sweeney, Beatrice M., 1951.
Culture of dinoflagellate Gymnodinium
with soil extract. Amer. Jour .Bot . ,
Vol. 38, No. 9, pp. 669-677.
Sweeney, Beatrice M., 195^.
Gymnodinium splendens, a marine
dinoflagellate requiring vitamin Big*
Amer. Jour .Bot., Vol. la,No. 10, pp. 821-8214-.
Thomas, W. H., and Krauss, R. W. 1955.
Nitrogen metabolispn in Scenedesmus as
affected by environmental changes .
Plant Physiol., Vol. 30, pp. 113-122.
Vallentyne, J. R. 1957-
The molecular nature of organic matter
in lakes and oceans, with lesser
reference to sewage and terrestrial soils.
Jour .Fish. Res. Bd. Canada, Vol. ll4-, No. 1,
pp. 33-82.
99
THE VERTEBRATES OF SCOPE
NOVEMBER 7 - DECEMBER l6, 1956
By
Robert Cushman Murphy 1/
American Museum of Natural History
My journal included observations on all
vertebrates except the small and larval
fishes taken in net hauls. Collecting of
sea birds from a skiff was undertaken at
oceanographic stations whenever weather
permitted, resulting in the acquisition of 50
specimens. The birds, collected primarily for
identification, have not yet all been studied
for subspecific determination. For the
purpose of this report specific status is in
most cases adequate. A later publication will
include data on taxonomy, habits, and stomach
contents.
FISHES
The bulk of the fishes captured, and now at the
Scripps Institution, are outside my province.
The following notes are restricted to larger
or otherwise readily observable species.
Ginglymostoma cirratum. Nurse shark. An
example about 1.5 m. in length swam under and
around the skiff shortly after daybreak of
Nov. Ik, k^> miles SW of Acapulco (surface water
29.6°C).
Prionace glauca. Blue shark. Observed several
times in the "Dome" area, S of latitude 10°N
and 200 or more sea miles W of Costa Rica. On
Nov. 22, at 09°05'N, 89<>3C0 W, a young example
only 65 cm. in length rubbed persistently
against the flanks and bottom of the skiff
until it was hauled aboard by the tail
(surface water 25.7°C).
Carcharias. Sharks of this genus or type
were observed almost daily in tropical waters.
They frequently assembled around the Stranger
when she was on station. In common with
certain other oceanic fishes, they were
strongly drawn toward any flotsam large enough
to cast a shadow. Bamboo poles and glass net-
floats proved sufficient to serve as an
attraction.
On Nov. 22, in the position noted under the
foregoing species, a Carcharias 2 m. in length
rubbed and banged the skiff for ten minutes,
sweeping its tail along the gunwale and
splashing showers of water all over the craft.
Such behavior is sometimes interpreted as an
effort to remove ectoparasites but it may
represent merely a thigmotactic drive.
Galeocerdo tigrinum. Tiger shark. On Nov. 16,
at 12" V71 N, 94°15' W, about 215 miles Off the
head of the Gulf of Tehuantepec, a young and
spotted example, less than 2 m. in length,
clung for some time to the vicinity of the
skiff (surface water 28°C).
Mobula
7*
Jumping ray. On Nov. ik, at l6°l6' N,
100°27' W. roughly k5 miles SW of Acapulco, a
ray about 60 cm. in lateral extent jumped and
somersaulted six times just ahead of the skiff
(surface water 29.6°C).
Other rays, not identified, were frequently
seen during the cruise.
1/ Dr. Murphy's participation in the cruise
was made possible through a gift to the American
Museum of Natural History from Mr. Edgar J. Marston
of La Jolla, together with a grant from the Council
of the Scientific Staff of the Museum.
Manta birostris. Giant ray; manta. The
specific name may possibly be open to question,
but the ray was indistinguishable in the field
from the Atlantic form.
101
The manta was encountered seven or more times.
The northernmost record was on Dec . 1^, about
60 miles W of Punta San Juanico, Baja
California (surface water less than 23°C).
The southernmost was in the shore waters of
Cocos Island, where two followed or kept in
close touch with the skiff for fully half an
hour . These approached within oar ' s length
and the larger was at least km. in breadth
(surface water 26.1*°C). The dorsal aspect of
their upcurled fin-tips, as seen in the air,
was blackish, but as soon as they submerged a
meter or more the color reflected through the
water became a pale tan, extraordinarily
reducing visibility.
Coryphaena hippurus. Dolphin: dorado.
Commonly encountered nearly everywhere S of
15°30' N, where on Dec. 13 the surface
temperature was below 23°C. Numerous dolphins
were captured. Their stomachs contained squids,
flying fishes, and parasitic nematodes.
The largest example measured 150 cm. in
standard length and was taken at 09° l6' N,
89°l8' W, on Nov. 20 (surface water 25.3°C).
The position is in the "Dome" area, 200 miles
W of Costa Rica. With this and with two other
dolphins I confirmed observations made by
Benjamin Franklin in the Atlantic Ocean on
Sept. 2, 1726.
Franklin's account, which is unknown to most
ichthyologists, relates to his first return
from England to Philadelphia. His Journal
states:
"We caught a couple of dolphins and fried them
for dinner... These fish make a glorious
appearance in the water; their bodies are of
a bright green, mixed with a silver colour,
and their tails of a shining golden yellow;
but all this vanishes presently after they are
taken out of their element, and they change
all over to a light grey. I observed that,
cutting off pieces of a just-caught, living
dolphin for bait, those pieces did not lose
their lustre and fine colours when the
dolphin died, but retained them perfectly."
The repetition of this experiment showed that
skin overlying severed chunks of myomeres
from the back remained dark blue after the
same area on the dying fish had turned almost
white. In like manner, sections from the
belly retained their pristine silvery yellow
hue, with pale blue spots, after the same part
on the body of the fish had faded.
The dermal chromatophores are under combined
control of hormones and the parasympathetic
nervous system. Proximal severing of the
fibers evidently leaves the hormonal influence
unopposed.
Exococoetidae. Flying fish. Observed
throughout the cruise. An example of
Cypselurus calif ornlcus flew aboard Stranger
during the night of Nov. 7 at 30°3U'N, not far
from San Diego, and another, kO cm. in
standard length, on Dec . 15, just N of Cedros
Island, Baja California. The surface water
at the first of these localities was 19.1°C.
Flying fish were conspicuous above the bank
surrounding the Alijos Rocks, Nov. 95 off the
Gulf of Tehuantepec, Nov. 16; and at 09° 1+6' N,
93°30' W, 300 miles from the continent,
Dec. 6. Weather had much to do with
observation because the fishes emerged most
actively during strong winds.
An example of Cypselurus nigricans, l8 cm. in
standard length, flew aboard within sight of
the Gulf of Dulce, Panama, on Nov. 25 (surface
water 26. VC).
REPTILES
Pelamis platurus . Sea snake. The northern-
most specimen was taken under a light in the
evening of Nov. 15, at about lk° N, 96<,10' W.
This was in a zone of upwelling to leeward of
the Gulf of Tehuantepec. The surface
temperature was only 25°C, whereas a few hours
earlier and to the north it had been 29°C .
Another was captured Dec . 1 in the Gulf of
Panama (surface water 27°C).
Sea snakes were most conspicuous off western
Panama, near Coiba Island, on Nov. 25, as
many as ten at once sometimes being within sight
The distribution of this 6pecies, the only
sea snake along the Pacific coast of America,
is graphically correlated with the major
oceanic circulation. The normal range extends
from no more than latitude 02°S (or even
nearer the equator, at La Plata Island,
- 102 -
Ecuador) northward to about 23°N, at the
mouth of the Gulf of California. Seasonal
countercurrent development sometimes leads
to a slight transgression of these limits but
the range is, in any case, latitudinally
asymmetrical, like that of many other marine
organisms inhabiting the warm zone between the
Peru and California currents.
such as off the semiarid Pacific coast of
Mexico, turtles may offer birds the commonest
and most used resting "islets". We repeatedly
saw boobies of two species, as well as certain
other birds, perched upon their backs.
Evidently such stowaways do not incommode the
surfaced turtles. This matter is referred to
further in the account of the birds.
Chelonia. Sea turtles. Turtles sighted
during the cruise of Stranger probably
included four species, namely Chelonia mydas
(green) Eretmochelys imbricata (hawksbill),
Caretta caretta (loggerhead), and
Lepidochelys olivacea (Pacific Ridley) . The
last two, both of loggerhead type, were
undoubtedly the commonest, and the only turtle
captured and certainly identified was
Lepidochelys . Identification of turtles in
the water at various distances offers
difficulties because of the changes in the
margin of the carapace that take place with
age and growth.
The example of Lepidochelys olivacea was
taken on Nov. 23 at 09°^1' N, 89°Mt-' W,
about 220 miles from the nearest land. Its
carapace was 51 cm. in length. On the left
side of its snout it bore a large barnacle,
not yet identified.
All other examples seen from shipboard had
best be listed merely as "turtles" . They
were noted as especially abundant on seven
different days of the cruise, namely
Nov. 13, ll+, 16, 23, 25, and Dec. 1 and lk.
Turtles were seen also on many additional
days in both coastal and (ff shore areas
between latitudes 26° N and Ok" N. The total
range of surface temperatures throughout
these waters and dates was 2^.8° to 29.7°C.
Sea turtles, like many oceanic fishes, show
great interest in flotsam. I repeatedly saw
them change course to approach the skiff or
one of the ship's floating bamboos supporting
a radar reflector . They would then nudge or
rub against the hard objects for long periods.
My most interesting observations concern the
ecological importance of turtles as resting
places for sea fowl. In seas of sparse flotsam,
BIRDS
Gavia immer. Loon. One seen Dec. 16 off the
coast of Baja California near the United States
border .
Fulmarus glacialis rodgersi. Pacific fulmar.
A gray-phase female collected Dec. 12 at
23°31' N, 111822' W. This is 30 miles offshore,
halfway between Santa Margarita Island and
Cape San Lucas (surface water 2^°C) . The
specimen probably represents the southernmost
record of the fulmar in any ocean.
Puffinus creatopus. Pink-footed shearwater.
Observed, always singly, on six days during
the cruise between the coastal waters of
southern Baja California and the vicinity of
Cocos Island. A casual representation of this
southern-hemisphere breeder north of the equator
during the normal nesting season is to be
expected.
Puffinus griseus. Sooty shearwater. A case
akin to that of the preceding species. Single
birds twice noted, once on Nov. 13, off
southern Mexico (surface water 29.7°C), and
again on Dec. 13, off Cape San Lazaro, Baja
California.
The sooty shearwater nests in the antiboreal
zone and usually passes rapidly across the
tropics on its migrations between higher
latitudes of the opposite hemisphere.
Puffinus gavia opisthomelas . Black- vented
shearwater. Many seen feeding, in company with
other petrels and terns, about kO miles off
Punta San Telmo, SE of Manzanillo, Mexico,
Nov. 13. The birds were in a natural oily
"slick" on the ocean.
On the return voyage, when we were bound north-
- 103
ward from Sebastian Yiscaino Bay and were
within sight of the San Benitos Islands,
where this shearwater nests, scores came
close to the ship on Dec. 15.
This is a "fluttering" shearwater, an apt
vernacular name originating in New Zealand
from where the topotypical race was described.
Other subspecies inhabit the Mediterranean
Sea.
Puff inus puffinus auricularis. Townsend'6
shearwater. Distinguishable from the
preceding species chiefly by its style of
flight, rather than by the blacker shade of
its dorsal surface, this shearwater was seen,
in company with wedgetailed shearwaters,
near 09*1*6' N, 93°30' W, on Dec. 6.
Townsend's shearwater is a weakly marked
race of the European Manx shearwater. The
specific distribution is cosmopolitan.
lerminieri subalaris. Galapagos
shearwater. One of the surprises of the
cruise was the abundance and wide
distribution at sea of the Galapagos race
of Audubon's shearwater. As a species,
srniinieri has world-wide tropical range.
The subspecifics character of subalaris
are strongly marked, notably in the
corneous nature of the nasal tubes.
Identification was confirmed by the
capture of an adult male with greatly en-
larged gonads at 11*13 • N, 90*55' W, on
Nov. 17. The position is south of the
Guatemala-Salvador boundary, nearly 150
miles from land.
This was the only Procellari-form bird that
showed curiosity, or what might be called
a "playful" interest, in the vessel. On
many occasions single birds or groups
performed swift and repetitious flight
maneuvers around the craft. The Nov. 17
example was one of the two that flew close
to the bulwarks of Strang- r many times at
dizzying speed, enabling me to make several
not too successful photographs.
Thereafter we saw these small shearwaters
on many dates along our course. They were
abundant off the coast of western Panama on
Nov. 2U-25. During the night of Nov. 30,
when we were bound outward through the Gulf
of Panama toward Cape Mala, hundreds of them
fluttered about within range of the ship's
lights, and after dawn of Dec. 1 seven of the
birds in a compact group flew up from astern
many times, swept to within arm's reach of the
rail, and then swung off widely to drop astern
and come up again.
We last saw this species near Cocos Island,
and in waters toward the NW, Dec. 1-6. The
northernmost records were in the neighborhood
of 11* N. Although the Galapagos Islands are
still the only known breeding grounds, it is
quite possible that this shearwater may prove
to be also a resident of Cocos Island.
Dr. Bell Shimada and other members of the
scientific group on Stranger informed me that
on an earlier cruise of the M/v Spencer F. Baird
in these same waters during late November or
early December large numbers of shearwaters,
which they believed to be this form, had
descended on the decks, sung in pairs, and even
copulated. The men had to toss the birds into
the air to get rid of them. The identification
is almost assuredly correct because no other
shearwater of the area would be at the peak of
its reproductive cycle at this season. Other
instances are known in which petrels in a
breeding state have adopted ships as convenient
"islands".
Puffinus pacif icus chlororhynchus . Wedge-
tailed shearwater. Or. the American side of the
Pacific this petrel nests only at the
Revillagigedo Islands. It ranges southward
through the warm ocean waters to the Pacific
coast of Colombia, where I collected specimens
on the Askoy Expedition.
Aboard the Stranger the following was observed
on three dates: a flock on the morning of
Nov. 17, near 11°N, 90°55' W; many, all of the
white-breasted phase, on Dec. 6, at 09°^' N,
93°30' Wj and birds of both dark and light-
breasted plumage phases on Dec . 16, off northern
Baja California (surface water circa 20°C).
Now and then wedge-tails crossed the bow of the
ship at close range, but never when collecting
proved possible.
Oceacodroma tethys kelsalli. Galapagos storm
petrel. Seen frequently throughout the cruise
and represented by six specimens in the
- 101*- -
collection. These establish the race as that
breeding at the Galapagos Archipelago. A
slightly smaller form nests on islets along
the coast of Peru.
Two males at the peak of breeding condition
were taken on Nov. 10 at 22°57' N, 113,3^' V,
a few miles N of the isolated Alijos Rocks
and nearly 200 miles W of Cape San Lazaro,
Baja California. Thereafter the species was
encountered all along our course to the "Dome"
area, Nov. 19-23. We met it again outside
the Gulf of Panama, in the waters around
Cocos Island, and for a thousand miles toward
the NW until Dec. 8. The range of surface
temperatures throughout these areas and dates
was 2l*.l° to 29.6°C.
Oceanodroma leucorhoa. Leach's petrel. Two
specimens collected, but which of the four
subspecies currently recognized along the
Pacific coast of North America they represent
has not yet been determined. It is likely
that the typical and most northerly race
migrates farther southward than any of the
other three.
The pattern of distribution of Leach's
petrel during the cruise of Stranger closely
matched that of the preceding species. The
first specimen flew aboard S of the Alijos
Rocks during the night of Nov. 9. Thereafter
the species was logged on Nov. 10, 11, 19, 20,
21-23, to the "Dome" area. Later, Dec. 2-1*,
we found it at 0V09' N, 83*31+' W, in waters
around Cocos Island, and for about 200 miles
northwe stwar d .
Probably surface temperatures have little
significance in relation to the winter
distribution of this storm petrel.
Loomelania melania. Black petrel. This
species nests at Los Coronados and San
Benitos islands W of Baja California and at
islands of corresponding latitudes within
the Gulf of California. It migrates south-
ward to the ocean off northern Peru, but
avoids the cool waters of the Peru Current .
Its winter range S of the equator appears to
be determined, indeed, by surface temperatures
dependent upon current-countercurrent controls ,
The black petrel was logged very frequently
between Nov. 11, at 21c07' N, 109*56 ' W
(S of the Alijos Rocks) to western Panama
and the Gulf of Panama. Later we found it at
our southernmost station (0V09' N, 83*31*' W),
around Cocos Island, and N toward the continent
to the latitude of Cape San Lazaro on Dec. 13.
The amplitude of surface temperature among all
the observations was about 2l*°C to 29.6°C.
Five specimens were collected. They had heavy
deposits of subcutaneous fat, as befits birds
in the early stages of a long migration. In
fact, they were the fattest of the four species
of storm petrels obtained on the expedition.
A female shot on Nov. 14, at l6°l6' N,
100*27' W, had her stomach and gullet crammed
with lantern fishes of a uniform 1*0 -mm. length.
Of all Pacific storm petrels of my personal
acquaintance, this one is most persistently
given to following vessels. The birds
accompanied Stranger for days on end, sweeping
widely across the wake and apparently
profiting from the artificial turbulence of
the water rather than from food cast overboard.
Halocyptena microsoma. Least petrel. Like the
preceding species, this tiniest of petrels
nests in the Mexican Pacific and Gulf area and
migrates southward into equatorial waters.
It was observed and occasionally collected along
our course between Nov. ll+ and Dec. 16. There
were periods of days in which none was seen,
but these appeared to have nothing to do with
latitude or with distance from the continental
coast . The northernmost record was near Cape
San Lazaro, Baja California, Dec. ll*. Surface
temperatures on all days on which the species
was noted ranged from about 2V to 29.6°C.
Phaethon aethereus . Red-billed tropic -bird;
boatswain-bird. Found in abundance at the
Alijos Rocks (21**57' N, 115*1*3' W), Nov. 9,
and casually observed in both coastal and
pelagic waters on eight other days, as far N
as Cedros Island, Baja California.
Alijos Rocks, approximately 185 miles W of
Cape San Lazaro, seem to have been bypassed by
105
the many ornithologists who have visited
islands W of Mexico. I find no mention
of them in availahle texts. They prove,
however, to he the probahle northernmost
breeding station in the Eastern Pacific
of three wide-ranging tropical ocean birds,
namely this species, the masked booby, and
the American man-o'-war bird.
The tropic-birds at Alijos Rocks were en-
gaged in active courtship, twos and threes
joining in swift pursuit flight and keeping
up an excited trill of their boatswain
whistles. A male collected wa6 at the
physiological peak of breeding. It dis-
gorged a 25-cm. fish (Colalabis saira) .
Surface temperatures in the neighborhood
of the Alijos Rocks were as low as 20.6°C.
The tropic-birds appeared to be nesting on
each of the three stacks of the group.
Pelecanus occidentalis . Brown pelican.
This is a continent-hugging species, of
little interest in an oceanographic campaign.
It has reached only one group of remote
oceanic islands- the Galapagos - where the
resident colony is isolated and racially
endemic .
We encountered two subspecies, californicus
of the northerly and relatively arid coast,
and carolinensis of the moist tropical
Middle American coast of both Atlantic and
Pacific. Nothing was learned about dis-
tribution boundaries or possible inter-
gradation of these two forms.
Sula dactylatra . Masked booby. This
largest of the tropical pelagic boobies is
to a great extent a flying fish-eater. White
adults, dark young, and birds in transitional
plumage were seen regularly after we had
reached the newly discovered nesting station
at Alijos Rocks. The species avoids forested
islands and continental coasts. It was not
present at Cocos Island, for example, although
common enough over the surrounding ocean
within a distance of a few hours' sail.
Discovery of the Alijos colony, where breeding
boobies appeared to be confined to the eastern-
most of the three stacks, rounds out our
knowledge of the nesting stations in the
Eastern Pacific. These extend from the
Alijos Rocks S to San Ambrosio Island, off
northern Chile, and include Malpelo (Colombia)
and La Plata (Ecuador). The LaPlata colony
is the only one within sight of the continent.
Malpelo has by far the largest booby
population.
At sea this booby showed marked curiosity
regarding conspicuous flotsam such as our
skiff and the radar reflector above bottle-
floats. The birds would swoop around them
again and again, and even attempt to alight.
As noted above, the masked booby also makes
regular and prolonged use of sea turtles as
rafts for resting on the ocean. Substantial
flotsam, such as logs, is used in the same
way, but it is likely that turtles offer the
mo6t plentiful opportunities for perching
throughout vast areas off soundings .
At any rate, on Nov. 15, some 90 miles off
the Gulf of Tehuantepec, I saw eight of these
boobies standing peacefully on turtles.
Again, on Dec. 8, much farther off the coast,
six more were observed resting in the same
manner. Lone turtle-perchers were noted on
numerous other occasions, in some instances
apparently sleeping, with the bill tucked
among the feathers of the back.
Sula sula. Red-footed booby. This is the only
tree-and shrub-nesting member of its family.
In breeding and feeding habits it occupies a
somewhat different ecological niche from other
boobies inhabiting the same area, thus avoid-
ing or reducing interspecific competition.
Like the masked booby, the red-foot is an
offshore and pelagic bird, rarely found near
continental coasts. We entered its strong-
hold only at Cocos Island, a well -populated
nesting station, and found it at sea only
within UO0 miles of that island, chiefly
toward the NW. It was the only booby that
followed the ship, played around the mastheads,
and alighted on the superstructure.
Approaching Cocos, one was caught on a fish-
hook. Others were collected at the island.
Throughout the tropical oceans this species
has several plumage phases, the taxonomic
106 -
significance of which is not yet well
understood. The Cocos Island population,
however, comprises only uniformly grayish-
brown birds, and we saw no other type on our
voyage .
Sula leucogaster brewsteri. Brewster's
booby. The case of this booby and the next
poses interesting biological and bio-
geographical problems. Both are subspecies
of the cosmopolitan brown booby, and both
are confined to the west coast of America
and outlying islands . From the topotypical
brown booby the two races differ in a
similar manner, notably in that the heads of
adult males have pale or whitish feathering.
The physical distinctions between the sub-
species brewsteri and etesiaca are slight
but are constant and readily recognized.
Physiologically, however, the differences
between these two races may be relatively
profound because brewsteri lives in an area
of high aridity, whereas etesiaca extends
from some unknown point N of western Panama
southward to the coast of Colombia. It
includes also Cocos Island. Whether there
is a hiatus between the coastal ranges of the
two races is yet unknown.
We first met Brewster's booby on Nov. 12,
midway off the mouth of the Gulf of
California. The birds were flying in pairs
or in groups of three. Next day the first
specimen was collected. Thereafter examples
were observed, sometimes standing on the backs
of turtles, as far as waters off the Gulf of
Tehuantepec . On the return voyage we saw this
booby again off the entrance of the Gulf of
California on Dec. 10.
Sula leucogaster etesiaca. Columbian booby.
The presence of boobies of this type in the
"Dome" area was inconclusive because of the
difficulty of discriminating, without
specimens in hand, between etesiaca and
brewsteri.
When we approached the coast of western Panama,
large flocks of Colombian boobies became a
familiar sight. Sometimes they were feeding
with other sea birds, such as cormorants,
Jaegers, and terns. A particularly large
concentration was passed on Nov. 25 off the
Islas de Ladrones, where they doubtless nest.
Later we found them in the Gulf of Panama and
along our course toward Cocos. On Dec. 1 a
female in breeding state was collected at
05°59' N, 79°l48' w.
While approaching Cocos on Dec. 3> we met a
movement of Colombian boobies 50 miles from the
Island. At Cocos they were nesting principally
on the outlying islets, particularly on
Manuelita or Nuez, where their nests, with
eggs and young in all stages, were underneath
tall shrubs in which red-footed boobies were
nesting. Although confined to the ground for
nidif ication, the Colombian boobies perch
freely on good-sized branches of trees, but
perhaps never on twigs.
The surface water at Cocos proved of slightly
lower temperature than that in the range of
brewsteri. far northward.
Phalacrocorax penicillatus. Brandt's
cormorant. The cormorants are all coastbound
birds in the part of the world under
consideration. This species was noted along
the coast of Baja California.
Phalacrocorax olivaceus. Bigua cormorant.
Observed in western Panama and in the Gulf of
Panama.
Phalacrocorax pelagicus. Baird's cormorant.
Although named pelagicus, this species is also
confined to the narrow continental platform.
It was noted only within a few miles of San
Diego and the Coronados Islands.
Fregata magnificens. American man-o'-war bird.
This species is common to both Atlantic and
Pacific sides of tropical and subtropical
America but, except at the Galapagos, it is
replaced by the following species as an off-
shore bird in the Pacific. Our most seaward
records were made near the breeding station of
Alijos Rocks, on Nov. 9. This is presumably
the northern limit of the nesting range on the
Pacific coast. Two adult males were collected
here.
Thereafter we saw this species regularly to the
Gulf of Panama, always interested in
107 -
aggregations of other sea birds and of fish and
porpoises at 1he surface. On the voyage to
the Cocos Island we left it far behind, but
picked it up again off Cape San Lazaro,
Baja California, on Dec. 13-
Fregata minor . Pacific man-o'-war bird. This
species; sometimes called the greater man-o'-
war bird (although it is smaller than
magnif icens) , occurs also in the Indian and
South Atlantic oceans. It nowhere reaches
American continental shores.
Both magnificens and minor, however, occur at
the Galapagos Archipelago, although perhaps
never at the same island. It has long been a
question as to which species is resident at
Cocos, a matter not solved until the visit of
Stranger.
On Dec. 3, we were met by scores of F. minor,
all in immature plumage, some tens of miles
E by S of Cocos. They mingled with our
escort of red-footed boobies, both astern
and circling the masts.
The adults at Cocos Island mostly soared high
above the hills and treetops. At times one
would swoop toward the water to harry a food-
laden booby. This ultimately enabled me to
shoot an adult breeding male. The species
was not seen elsewhere.
Casmerodius albus egretta. American egret.
At noon on Dec. 1 one flew, out of gunshot,
past my skiff at 05°59' N, 79° >*8' W. The
position is on the open ocean about 9° miles
S of Cape Mala, Panama.
Anas platyrhynchos . Mallard. A female duck,
apparently a mallard, alighted and then took
o-ff from the ocean, close to Stranger, on
Nov. 20, at 09°l6' N, 89°l8' W, about 200
miles off the Costa Rican coast. On Nov. IT,
12 similar ducks passed high above us at an
equal distance from the nearest land
(El Salvador) .
Aythya af finis, Lesser scaup. Not observed
at sea, but on Nov. 28 a flock took off from
Gatun Lake, Canal Zone, in front of the
Barro Colorado Island Laboratory. The species
has apparently not previously been recorded from
Barro Colorado.
Phalaropus fulicarius. Red phalarope.
Observed along course, both near the coast
and far offshore, between Nov. 19 and
Dec. 1^. The northernmost record was
off Point San Juanico, Baja California.
Red phalarope6 were usually met with either
in pairs or in small flocks. One was once
6een standing on the back of a turtle. Our
only specimen, which was extremely fat, was
taken on Dec. 8 at 1V37' N, 100° 09' W.
Lobipes lobatus . Northern phalarope.
More abundant than the foregoing species
and likewise usually found in either pairs
or flocks. It still ranged as far north as
the ocean off San Diego on the last day of
our voyage, Dec. l6. One was collected off
the Gulf of Tehuantepec on Nov. 15.
Both species of phalaropes showed a
predilection for oily "slicks" on the
ocean.
Stercorarius pomarinus . Pomarine jaeger.
The commonest of its family throughout the
cruise. Seen everywhere, and almost daily,
between San Diego and Panama, and in the
waters NW of Cocos Island. In the Gulf of
Panama it was parasitizing the laughing
gulls. An example was collected on Nov. 1^.
Stercorarius parasiticus. Parasitic jaeger.
Less common than the pomarine jaeger, but
presumably as widely distributed. A
specimen collected to represent this species,
however, has proved to be the next.
Stercorarius longicaudus. Long-tailed jaeger.
A very young, practically fledgling, male
jaeger, shot on Nov. Ik at l6°l6' N,
100*27' W, has turned out to be longicaudus.
The species was not noted elsewhere.
Catharacta skua chilensis. Chilean skua.
Two brightly cinnamon skuas, seen at close
range from Stranger on Nov. 15, over a
"slick" off the Gulf of Tehuantepec, assuredly
were of this form, with which I became well
acquainted in Peru and Chile. The skuas
were in company with Sabine's gulls, boobies,
and storm petrels .
- 108 -
Larus occidentalls . Western gull. Seen
wealth-ward from San Diego to a distance
of 90 miles off the Gulf of Tehuantepec.
Along the shores of Baja California this
species appeared in alternate with bands
of the California gull.
Larus calif ornicus. California gull.
Encountered only along the coast of Baja
California. Off the broad entrance to the
Gulf of California, Dec. 10-12, we met
many immature examples which behaved like
veteran pensioners of ships and were
content to wait hours for jettisoned
garbage. At one time I counted 250 around
Stranger. White, adult California gulls
were mostly seen farther northward.
Larus atricilla. Laughing gull. Common in
the Gulf of Panama. First seen in
considerable numbers immediately after we
had rounded Cape Mala on Nov. 26. On Dec. 1
several followed us out to the high sea for
about 100 miles S of Balboa. This gull was
also common in Gatun Lake, Canal Zone,
mingling with small flocks of black terns.
The first specimen of the laughing gull,
however, was collected far off the coast
of Baja California, at 22°57' N, 113°3^' W,
on Nov. 10. A second was taken at
05°59' N, 79°W5' W, Dec. 1.
Larus pipixcan. Franklin's gull. Adults
still wearing full summer plumage were seen
off western Panama on Nov. 25. The two
collected were both young birds, taken at
sea on Nov. 23 at 09°la' N, Q9°hh' W. The
position is in the "Dome" area, about 2*10
miles W of Costa Rica. The longitude, which
is far to westward of South America, passes
through the Galapagos Islands, to which
Franklin's gull is a regular winter visitor.
Larus heermanni. Heermann's gull. Observed
between Cedros Island, Baja California, and
San Diego, Dec. 15-l6.
Xema sabini. Sabine's gull. Seen
pccasionally and usually at long range,
southward to Panama. A female was collected
on Dec. 12 at 23°31' N, 111°22' W. The
position is W and a little N of Cape San Lucas.
Sterna hirundo. Common tern. Small terns
were seen on many dates, but the only certain
identification is based upon a female of this
species collected on Nov. 15 at l1v°17' N,
96°3l4.' W, off the Gulf of Tehuantepec.
Thalasseus maxlmus . Royal tern. Common in
the Gulf of Panama, Dec. 26-30.
Chlidonias niger . Black tern. First observed
off the Gulf of Dulce, western Panama,
Nov. 25. Common in the Gulf of Panama and on
Gatun Lake, where it mingled with laughing
gulls .
The black tern in its winter range clings
closely to tropical coasts and flotsam-filled
waters. It never dives and it rests mostly
on floating vegetation. Therefore it is
always most abundant where rivers flow to the
ocean through forested areas . It is very
rarely found out of sight of land.
Anous stolidus. Brown noddy. An adult
female with slightly enlarged ovaries was
collected on Manuelita Islet, off the northern
point of Cocos Island, on Dec. 3. On
Dec. 8 at 1V37' N, 100°09' W, about 190 miles
south of Acapulco, I saw a small flock of
this species.
Megalopterus mlnutus . Black noddy. Black
noddies came aboard Stranger early in the
morning of Nov. 23. Later in the same day
an adult male was collected at 09° kl1 N,
Qg'kk' W, which is in the "Dome" area, about
240 miles W of Costa Rica.
On Dec . 3 several examples were seen flying
in and out of a sea cave on Manuelita or
Nuez Islet, Cocos Island.
Land Birds. A considerable number of land
birds alighted on Stranger in various parts of
the cruise . Some of them could be only
approximately identified:
Speotyto cunicularia. Burrowing owl. Nov. 13,
more than 1+0 miles off Petacalco Bay, Mexico .
Large flycatcher. Nov. 25, W of Coiba Island,
Panama.
- 109
Empidonax . Small flycatcher. Nov. 9, at
the Alljos Rocks.
Hirundo rustica erythrogaster . Barn swallow.
Gulf of Panama, Dec. 26 and 30.
Petrochelidon pyrrhonota. Cliff swallow.
Nov. 11 at 21*07' N, 109°56' V, midway across
the mouth of the Gulf of California.
Hylocichla. Thrush (resembling a hermit thrush)
Nov. 2k, about 80 miles off the coast of Costa
Rica.
Vermivora peregrina. Tennessee warbler. A
young bird, sex indeterminable, came aboard
on Nov. 10 at 22°57' N, 113°3^' W.
Vermivora ruficapilla. Nashville warbler .
Severl flew aboard W of Coiba Island and off
the Gulf of Panama, Nov. 2k and 25. One
younger one of undetermined sex was found in
the ship's laboratory and was preserved.
Ammi
.odramus. Sparrow. Nov. Ik, l6°l6' N,
100° 27' W, off Acapulco.
MAMMALS
Zalophus calif ornianus. California sea lion.
About a dozen were on and around the middle
Alijos Rock on Nov. 9. The other two stacks
of this group could be scaled only by winged
creatures. Otherwise we saw sea lions only
at the Coronados Islands, and on the channel
buoys of San Diego.
Mirounga angustirostris . California sea
elephant . One adult bull seen swimming off
the northern end of Cedros Island, Baja
California, where there is said to be a
small colony.
Rhachianectes glaucus. Gray whale. One
surfaced near Stranger among the Coronados
Islands on Dec. 16.
Physeter catodon. Sperm whale. This
species was several times sighted at long
range and recognized by the character of
its spout.
On Dec. 6, near 09° k6' N, 93*30' W, nearly
1*00 miles west of Costa Rica, we sighted
three sperm whales and Stranger followed
them at reduced speed, finally approaching
within 30 m. Two cows and a calf lay side
by side. The adults were each about 12 m.
in length, and the calf, which stuck to the
left flank of its mother, seemed only three
to four m. shorter. All three whales
sounded together and came up a quarter-mile
to the right of their former course. Later
in the same day two more sperm whales were
watched at a distance of a mile or more .
Globicephalus . Blackfish. I have no way
of knowing whether the blackfish seen on
several occasions represented the species
sc amnion i or macrorhynchus . They appeared
not infrequently around the ship all the way
from northern Baja California to waters
outside the Gulf of Panama.
Delphinus bairdi. Porpoise. Schools of
porpoises, indistinguishable to me from
D. delphis Of the Atlantic, were presumably
this species. Large groups were
encountered as follows: Nov. 12, 19* N,
106° W, two schools; Nov. 17, 11*13 • N,
90°55' W; Nov. 2k, 08°te' N, 86° W;
Dec. 6, near 09*1*6' N, 93°30' W; Dec. lk,
off Point San Juanico, Baja California;
Dec. 15, E of Cedros Island.
In the evening of Dec. 1^, when porpoises
were showing great activity close to
Stranger, the EDO of the sonar equipment was
turned on to receive their communications.
Porpoises signal in a language of high
frequencies beyond the range of human ears.
But the EDO pulled this down to 8000 cycles
and the result was like a dawn chorus of
birds in May. Whistles, piping, chattering,
and musical squeals came out of the depths
in a cheerful medley.
Prodelphinus graf f mani . Spotted porpoise.
On Nov. 26 a school of porpoises,
indistinguishable to my eyes from P.
plagiodon of the Atlantic, accompanied the
vessel on two occasions in the Gulf of
Panama.
Mesoplodent whale. On Nov. 10, near
22° 57' N, 113 °3l+' W, which is about 125
- 110
miles SW of Santa Margarita Island, Baja
California, an unidentified mesoplodent
overtook and passed Stranger . It was
approximately 10 m. long and seemed to
have a pronounced neck constriction; it
produced no visible spout during several
rises.
Ill -
THE ALCOHOL-SOLUBLE AND INSOLUBLE FRACTIONS OF THE PHOTOSTNTHETICALLY
FIXED CARBON IN NATURALLY OCCURRING MARINE PHYTOPLANKTON POPULATIONS
by
William H. Thomas
Chemical analyses of phytoplankton cells may-
give information valuable in determining the
nutritive value of such cells to their
predators. Such analyses are most easily
carried out with laboratory-cultured cells,
but such cells may not truly represent those
occurring in nature. This paper reports
experiments made at sea with naturally
occurring populations in which determinations
of the alcohol-soluble fraction of marine
phytoplankton were made. These determina-
tions may give an indication of the gross
chemical composition of photosynthesizing
cells .
Because of the small numbers of algal cells
present per unit volume of pelagic water,
attempts to harvest these cells for purposes
of chemical analyses by the usual chemical
means would be extremely time consuming.
Fortunately the use of the C-^ technique of
labeling the organic matter produced by
phytoplankton greatly increases the
sensitivity of a chemical extraction pro-
cedure so that only relatively small volumes
of water need to be handled.
In these experiments a sample of surface
water was taken with a plastic bucket.
Aliquots of this sample were added to 250-ml.
ground-glass-stoppered bottles, C^^was added
to each bottle, and the bottles were
illuminated at 1200-1^00 foot-candles at the
surface sea-water temperature in a glass-
bottomed water bath. After incubation,
aliquots of the water were filtered through
HA Millipore filters (0 .^5 H pore size) to
determine the total activity fixed. Another
aliquot was filtered through a sintered-glass
filter. The residue on this filter was
extracted with boiling 80°/o ethanol. An
aliquot of the combined extracts was
evaporated on a steel planchet to determine
the extractable activity. A linear
relationship between volume of extract and
activity showed that self -absorption
corrections were not necessary when volumes
no larger than 0.50 ml. were evaporated on
the planchets. The proportion of extract-
able material in the cells was determined
by dividing the activity extracted by the
total activity in the cells. The assumption
is made that after this long period of
incubation all chemical entities in the
cells are labeled with Cl^ to the same
extent and that the ratio of extractable
activity to total activity truly represents
the proportion of alcohol soluble in all
materials. It is further assumed that no
losses of Cl^ occurred during evaporation
of the extract on the planchets.
Considering the speed of the photo synthetic
cycle as shown by Buchanan et _al. (1952),
(steady-state labeling of sugar phosphates
after about 15 minutes) the first assumption
seems reasonable (cf. also Fogg, 1956).
Compounds appearing in the extract would
include sugars, amino acids, sugar phosphates,
pigments (less their protein moieties) and
lipids. Substances remaining behind would
include proteins, polysaccharides, and
nucleic acids.
An initial preliminary experiment was
performed at Station 0-1, 50 miles west of
Baja California (30801' N latitude and
ll6°l*-9' W longitude). In this experiment
only four bottles were incubated (for eight
hours) and the activity recovered in the
extract was only 92 cpm. above background In
10 ml. of extract. The total activity on the
millipore filter was 619 cpm. Thus about
15°/o of the activity was extracted. Because
113
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115 -
of the difficulty in determining the
activity of an extract of such a low
specific activity, and because no
replication of the plating was made, this
result was only considered preliminary.
It served to establish the range of activity
to be expected in two further experiments.
The next experiment was performed at Station
S-9 in the Central American thermal anticline
80 miles off Costa Rica (9°28' N latitude,
89el8' W longitude) on November 21, 1956.
Sixteen 250-ml . bottles of surface sea water
were incubated with 2,887,000 cpm. of Clk per
bottle for eight hours, and at 1200-1^00 f c .
and 25°C. After incubation the contents were
pooled in a plastic bucket, and 250 ml. of
this pool were immediately filtered through a
Millipore filter (MP-1-1) to determine the total
activity fixed. Then 1750 ml. were filtered
through an F-porosity sintered-glass filter
(maximum pore size 5 u). Two 250-ml. portions
Of the filtrate from this last filtration were
then filtered through Millipore filters
(MP-1-2 and MP-1-3) to determine the portion
of the activity not retained by the glass
filter. The residue on the filter was
washed twice with non-radioactive sea water
and then was extracted four times with
boiling 80°/,, ethanol. The combined extracts
were dried with a hot-air stream at 35°C and
then taken up in 7.5 ml. of twice-distilled
water to yield extract E-l. This whole
process was then repeated on the rest of the
pool to give Millipore pads PM-2-1, MP-2-2
and extract E-2 which had a final volume of
5 ml. derived from 1750 ml. of pool. The
glass filter used in this last extraction
had a maximum porosity of 1.2 u. Two 0.50 -ml.
aliquots of each extract were evaporated on
steel planchets at 35°C under a hot-air
stream and an infrared lamp (E-l-1, E-1-2j
E-2-1, E-2-2). The results of these
experiments are shown in the Table 19-
The next experiment was performed at Station
S-10, 130 miles off Costa Rica (8°l*-2' N
latitude, 86°00' W longitude) on November 2k,
1956. Fourteen 250-ml. bottles of surface sea
water were incubated with 5,77^,000 cpm. of
Cl1*- per bottle for five hours at 1200-1^0
fc. at 26<>C. After incubation the contents
of one bottle (260 ml.) were filtered through
a Millipore filter (MP-3-l) • The contents of
six bottles (1580 ml.) were filtered through
an M-porosity sintered-glass filter
(maximum pore size 1*+ u). Two 250-ml.
portions of this filtrate were filtered
through Millipore filters (MP-3-2 and
MP-3-3). The residue on the glass filter
was washed and extracted in the same manner
as in experiments 1 and 2. The combined
extracts were made to lO.U-ml. final volume
without drying and re-extracting with water
to yield extract E-3. The whole process was
repeated with the seven remaining bottles to
yield Millipore pads MP-^-l, MPA-2, and
MP-1(— 3 and extract E-1* which was derived by
filtration of 1575 ml. of the original water
through an M-porosity glass filter and which
had a final volume of 7.6 ml. Two 0.50-ml.
aliquots of each extract were plated on
planchets as in experiments 1 and 2. The
results of these experiments are shown in
Table 20.
These experiments show that about 15°/o of
the cellular carbon in naturally occurring
phytoplankton populations is alcohol soluble.
A similar proportion of soluble to insoluble
material (expressed on a dry-weight basis)
is found generally in those few algae which
have been investigated (cf. Fogg, 1953). It
is striking that there is little variation
in the percentage of extractable material
in phytoplankton from the three areas .
Presumably nutrient conditions might be
different in the various areas and the cells
might reflect thi6 by having differing
proportions of soluble material.
It can be inferred from the data, if the
assumption e made that the alcohol-soluble
material is al60 soluble in sea water, that
when an algal cell dies 15° /o of the cell
material would be immediately released to the
water and would serve as food for bacteria and
other hetrotrophic organisms. A portion of
this material might also become a part of
that more resistent dissolved organic material
which accumulates in the ocean. It is also
probable that only 85°/o of the material
produced by phytoplankton has any chance at all
of reaching the bottom and becoming a part of
the organic material in sediments.
If a phytoplankton cell is eaten before it
dies, then some 15°/o of its material is
immediately available for incorporation into
the body tissues of the animal which eats it.
- 116
Some 85°/o of the cell material would have to
be broken down by digestive enzymes in the
gut of the animal before incorporation. Also
fecal pellets are probably wholly made up of
this insoluble material.
REFERENCES
Buchanan, J. P., J. A. Massham, A. A. Benson,
D. F. Bradley, M. Calvin, L. L. Dans,
M. Goodman, P. M. Hayes, V. H. Lynch,
L. T. Norris, and A. T. Wilson. 1952.
The path of carbon in photosynthesis XVII.
Phosphorus compounds as intermediates in
photosynthesis. In McElroy, W. D. and
B. Glass, editors. Phosphorus Metabolism,
Volume II, pp. MO- 459. Johns Hopkins
Univ. Press, Baltimore.
Fogg, G. E. 1956.
Photosynthesis and formation of fats in
a diatom. Ann. Bot . (N.S.), Vol. 20,
pp. 265-285.
Fogg, G. E. 1953.
Metabolism of Algae. John Wiley and
Sons, Inc., New York.
117 -
INT . -DUP . SEC . . HASH . . D . C . U 7 3 << 0
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