HALIFAX RIVER ALGAL ASSAY REPORT
BY
R. L. RASCHKE
D. SCHULTZ
ENVIRONMENTAL PROTECTION AGENCY
SURVEILLANCE AND ANALYSIS DIVISION
ATHENS, GEORGIA
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EPA 904/9-77-035
HALIFAX RIVER ALGAL ASSAY REPORT
BY
R. L. RASCHKE
D. SCHULTZ
Environmental Protection Agency
Region IV
Surveillance and Analysis Division
Ecology Branch
Athens, Georgia
October, 1977
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TABLE OF CONTENTS
PAGE NO.
SUMMARY 1
INTRODUCTION 2
STATIONS 4
METHODS 5
RESULTS 8
August Algal Assays 8
October Algal Assays 10
DISCUSSION 15
LITERATURE CITED 21
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LIST OF FIGURES
FIGURE PAGE NO.
1 Halifax River, Valusia County, Florida 23
2 Algal Assay, Maximum Yield, August, 1976 24
3 Algal Assay, Maximum Yield, October, 1976 25
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LIST OF TABLES
Page No.
1 Halifax River Salinity, August, 1976 ........... 26
2 Nutrient and EDTA Additions to Halifax River Water
Samples, August, 1976 .................. 27
3 Nitrogen Additions to Halifax River Water Samples,
October, 1976 ...................... 28
4 Average Maximum, Yields (mg dry weight/liter) , Halifax
River Water Samples, August, 1976 ............ 29
5 Comparisons of Average Maximum Yield Within Stations,
August, 1976 ....................... 30
6 Chemical Analyses of Halifax River Water Samples in
mg/liter ......................... 31
7 Comparison of Average Maximum Yields and Chemical
Analyses of Halifax River Water Samples, August, 1976 . . 32
8 Average Maximum Yields (mg dry weight/liter) , Halifax
River Water Samples, October, 1976 ............ 33
9 Comparison of Average Maximum Yields Within Stations,
October, 1976 ...................... 34
10 Average Maximum Yield of Dunaliella (mg dry weight /liter)
in October, 1976 Nitrogen Spiked Halifax River Water
Samples ........................ , 35
11 Predicted and Actual Average Maximum Yields (mg dry weight/
liter) for Dunaliella October Halifax Control and Added
Nitrate Spikes ...................... 36
12 Dunaliella dry weight (mg) produced per unit of nitrate-N
at day 14 in defined medium ............... 37
13 Acceptable and Actual Levels of Nutrients for Halifax
River mg/liter (FDER Criteria) .............. 38
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SUMMARY
Algal assay methods developed by EPA were used in this investigation
as a result of concerns of Valusia County, Florida about population influx
and pressures in the Halifax River Basin, and to aid the Water Programs
Division in its evaluation of waste load allocations. An explanation of
the theory and practice of algal assay technique is provided in the text,
and the findings of this investigation are presented below.
1. Chemical data, low N:P ratios, plus algal assays indicated that
nitrogen was the limiting nutrient in Halifax River waters and
that the test alga was more sensitive to nitrogen inputs either
alone or in combination with other nutrients.
2. Phosphorus was in excess and the test alga did not significantly
respond to phosphorus spikes as it did to nitrogen spikes. Phosphorus
concentrations in Halifax waters were greater than acceptable levels
for Florida waters.
3. Greatest growth and high sensitivity to 0.2 mg nitrogen/liter spikes
were observed at stations located in the vicinity of sewage treatment
discharges.
4. Algal assay data indicate that at stations near large sewage discharges
potential algal nuisance problems could occur with the addition of 0.2 mg
nitrogen per liter (0.2 grams per m^).
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INTRODUCTION
The Halifax River is a part of the Intracoastal Waterway in Valusia
County, Florida (Figure 1). It flows in a southerly direction receiving
freshwater inputs primarily from the Tomoka River and Halifax Creek. Oceanic
waters enter at the southern end through Ponce de Leon Inlet. According to
Harvey (1) the Halifax River is flushed by tides as far north as 1.5 miles
south of the Port Orange Causeway (Figure 1) (2.5 miles north of the inlet)
and severe deterioration of water quality would not be expected. This cause-
way probably restricts water movement as evidenced by the 10 ppt difference
in salinity downstream and upstream from the Port Orange Causeway (Table 1).
The Port Orange Causeway as well as several other bridges increase residence
time for waters entering the northern portion of the river (1).
A number of municipal treatment plants discharge wastes into the Halifax
River. Holly Hill, located near Station 24 (Figure 1), discharges about 0.60
mgd of treated wastes into the Halifax River (1). Two sampling stations (9AA
and 8AA) (Figure 1) are in the vicinity of the Daytona Beach Sewage treatment
plant which discharges 12.0 mgd. Station 11 (Figure 1) is near the Port Orange
sewage treatment plant which discharges approximately 1.0 mgd into the Halifax
River (1).
Population influx and pressures in the Halifax River Basin, including
Valusia County, have necessitated an evaluation and allocation of waste
loads entering the Halifax River and its tributaries. As a result of
Valusia County's concerns, a program was initiated to gather additional infor-
mation for the Dynamic Estuary Model (DEM) used by the Water Programs Division
of EPA, Region IV to predict water quality conditions and aid in determining
waste load allocations.
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As a supplement to modeling efforts, algal assays were conducted
at EPA's Athens, Georgia laboratory to:
(1) determine which nutrients—nitrogen, phosphorus, micro-
nutrients—limit average maximum algal yield (growth) in
the Halifax River samples.
(2) determine how sensitive samples are to changes in the
limiting nutrient concentration.
The algal assay is based on Liebig's law of the minimum which states
that "growth is limited by the substance that is present in minimal quantity
in respect to the needs of the organism." The marine algal assay has been
designed to assess receiving waters of varying salinity as to nutrient
status, biostimulation potential, and sensitivity to change in nutrient
concentration.
When large amounts of nutrients are available, excessive algal growth
can occur; both the abundance and composition of algae are related to water
quality because algal growth is influenced by the availability of nutrients.
The design, evaluation, and applications of algal assays have demonstrated
the ability of unialgal assays to identify and assist in the management of
water quality problems.
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STATIONS
Water samples were collected for algal assay determinations at
the following locations:
Station Description
32 Located in Tomoka Basin at the upper
and of Halifax River.
24 Located about midway between Holly Hill
and Ormond Beach near channel markers 22
and 24.
9AA Located adjacent to the city of Daytona
Beach and due east of channel marker 33.
8AA Located adjacent to the city of South
Daytona and due west of channel marker 50.
11 Located midway between the city of Port
Orange and Ponce de Leon Inlet and due
east of channel marker 69.
2 Located upstream from New Smyrna Beach
and east of channel marker 30.
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METHODS
Water samples were collected on August 18, 1976, by the contractors,
Briley, Wild and Associates and Russell & Axion, at mid-depth every 6 hours
over the 24 hours during a period of two tidal cycles. Samples were composited
for each station and sent refrigerated in clean polyethylene "cubitainers" to
the Ecology Branch in Athens, Georgia, for processing according to the method
found in EPA's Marine Algal Assay Procedure (2).
Samples were filtered through a 0.45-micron membrane filter, and
chemical analyses of the unfiltered and filtered water were conducted for
total phosphorus and nitrogen, including nitrate-nitrite, ammonia, and total
Kjeldahl nitrogen.
Algal assays for each station composite were conducted in triplicate using
40 mi each of filtered Halifax River water in 125-m£ Erlenmeyer flasks.
A series of nitrogen and phosphorus spikes (Table 2) were added to deter-
mine if Halifax River waters were limited in either or both nutrients.
The purpose of single nutrient treatment is to establish the identity
of limiting nutrients. Synthetic organic ligands such as EDTA as a treat-
ment are used to insure the availability of trace elements and test for
the presence of toxic metals which when in excess are inactivated through
chelation. Combined nitrogen and phosphorus additions were utilized
to investigate the possibility that both nutrients were present in Halifax
waters in amounts so small that additions of either nutrient alone would
not yield a measurable response. Sodium nitrate provided the nitrogen
source and dibasic potassium phosphate the phosphorus source to test
flasks. Flasks were inoculated with Dunaliella tertiolecta cells to
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give an initial culture of 102 cells/mA Inoculated flasks of
Halifax River water samples, without the added EDTA, phosphorus or
nitrogen, served as controls.
Cell counts were made with a Coulter ZB-1 electronic particle
counter equipped with a mean cell-volume (MCV) computer. Algal cells
were counted on Monday, Wednesday, and Friday of each week. Counting
continued until the maximum yield of each replicate flask was obtained.
Maximum yield was attained between 11-20 days. Cell counts were converted
to mg dry weight/liter according to the data reduction equation in the
Marine Algal Assay Procedure (2). The maximum dry weights of the repli-
cate flasks were averaged and are expressed as average maximum yield.
The August tests showed that nitrogen was limiting; therefore, a
series of increasing concentrations of nitrogen as sodium nitrate were
added to each station water sample collected in October. The October treatments
(Table 3) included spikes of 0.2, 0.4, 0.8, and 1.6 mg nitrogen/liter
to each of three sets of replicate flasks. Three untreated replicates
of Halifax River water were used as a control, and three replicates of
algal assay media (Treatment 1) were used as a reference check on cell
growth as in the August algal assays. Replicate flasks contained 104
Dunaliella tertiolecta cells/m.£ initially. Dunaliella was allowed to
grow under standard conditions (2) until maximum dry weight/liter was
attained. October yield data was expressed as in August. Some of the results
of the October algal assays have been omitted because examination of the data
revealed that these cultures were not inoculated with algal cells.
The greatest growth of Dunaliella grown in marine algal media
(Treatment 1) was 500 mg dry weight/liter; this is equivalent to
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4,168,355 cells/mi with an average MCV of 139. This growth level is
much greater than that found in nutrient-spiked Halifax River waters
where the greatest growth was 191.6 mg dry weight/liter. The average
cell count that produced 191.6 mg dry weight/liter was 1,246,168
cells/mS, with a MCV of 180. The growth level of biomass and number
of cells produced in marine algal assay medium indicates there was
no crowding of cells and no carbon limitation due to excessive growths
in any of the Halifax River water treatments.
Data were analyzed for significant differences by using the cumulative
t-test (10). The .15 probability level was used to detect significant
differences. That means there is a 15% probability of being wrong in
assigning a significant difference to the sample means being compared.
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RESULTS
August Algal Assays
During August algal assays an effort was made to determine whether
nitrogen or phosphorus was limiting algal growth. For the purposes of
discussion, we have grouped those Halifax River stations (24, 8AA and
9AA) which receive large amounts of sewage and are between causeways
into one category called "impacted stations." The stations (2 and 32)
which receive the least amount of sewage have been termed "end stations."
Growth in nitrogen-spiked flasks ranged from 43.7 mg dry weight/liter
(Treatment 5) at end Station 2 to 101.1 mg dry weight/liter (Treatment
5) at impacted Station 9AA, whereas growth in flasks not receiving
nitrogen additions ranged from 16.0 mg dry weight/liter (Treatment 6)
at Station 11 to 61.5 mg dry weight/liter (Treatment 6) at Station 9AA
(Table 4, Figure 2). Water samples treated with nitrogen alone, and in
combination with other materials, generally had a greater growth of algal
cells than any other treatment. The single exception was the Station 2
waters. However, greatest growth for all water samples was always from
a nitrogen-spiked treatment (Table 4, Figure 2) either alone or in com-
bination with other chemical additions.
Nitrogen-spiked sample yields (Treatment 4) were significantly
greater than control (Treatment 2) or phosphorus-spiked (Treatment 3)
sample yields except at Station 2 (Table 5). Variance in the Station 2
data may have masked true differences which were not detectable by the
t-test.
In a comparison of average maximum yields between phosphorus-spiked
samples (Treatment 3) and control samples (Treatment 2), significant
differences were detected in Stations 9AA and 8AA samples (Table 5)
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indicating that phosphorus additions to these two impacted stations
affected yields, but not as much as nitrogen did (Table 4).
Combined spikes of nitrogen and phosphorus (Treatment 5) were
included to investigate the possibility that both nitrogen and phos-
phorus were present in Halifax waters in amounts so small that additions
of either nutrient alone would not yield a measurable response. An
examination of Table 4 and Figure 2 shows this was not the case as a
measurable response was noted, especially in the nitrogen-spiked samples
(Treatment 4).
Synthetic organic ligands such as EDTA are commonly used to insure
availability of trace elements for algal growth in defined media and
to diminish toxic effects of metals through chelation. EDTA was
added to Halifax River water samples (Treatment 6) for the purpose of
detecting metal toxicity. Figure 2 and Table 4 show there was little,
if any, increase in growth when EDTA was added to Halifax River waters
(Treatment 6). In fact, there were no significant differences at the
.05 and .10 probability level between controls (Treatment 2) and Treatment
6 at all stations (Table 5). Only at the .15 probability level was
Treatment 6 significantly greater than the control at Station 9AA (Table
5); this indicates that some metal toxicity may be present at Station 9AA.
Generally, it does not appear that metals were available in sufficient
quantities to suppress algal growth in the Halifax River at the time
samples were collected.
Control growth ranged from 21.4 mg dry weight/liter at Station 8AA
to 49.8 mg dry weight/liter at Station 2 in August (Table 4). Station
9AA with the second highest average maximum yield of 44.5 mg dry weight/
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liter was the most sensitive station to equivalent nitrogen-spikes
(Treatment 4) because the addition of 0.2 mg nitrogen/liter increased
growth 50.3 mg dry weight/liter from 44.5 mg dry weight/liter to 94.8
mg dry weight/liter (Table 7).
The August algal assay data presented herein indicates that the
Halifax River is nitrogen-limited and much more sensitive in growth
response with additions of nitrogen alone or in combination with other
chemicals. The chemical data further verifies that the Halifax River in
August was nitrogen-limited; the N:P ratios (Tables 6 and 7) were all less
than 10, indicating nitrogen limitation. Based on the above facts, only
incremental increases of nitrogen were added to Halifax River waters collected
in October.
October Algal Assays
During the October water quality study, conducted by Briley, Wild and
Associates and Russell & Axion, samples were collected for algal assays
at the same stations as in August. Emphasis was placed on nitrogen
treatment by adding increasing amounts of nitrogen as NaNO~ to sample
waters.
Sample waters impacted by sewage inputs (Stations 24, 9AA, and 8AA
and to some extent Station 11) were quite sensitive to nitrogen additions.
There was a 2.7 to 9.1-fold increase in growth at these stations with
the addition of 0.2 mg nitrogen/liter (Treatment 3) (Table 8 and Figure 3).
All station waters except the end stations, Station 32 at the Halifax
River headwaters, and Station 2 near Smyrna Beach (Figure 1), had
significantly greater growth than the controls when 0.2 mg nitrogen/
10
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liter was added to sample waters (Table 9). With successive additions
of nitrogen, successive incremental increases were less noticeable
(Table 10).
At the end stations, the differences between the 0.2 mg nitrogen/
liter spike (Treatment 3) and control waters was not as striking as
the other four stations (Figure 3). At both end stations there was
generally a gradual increase in growth with each incremental addition
of nitrogen until average yields of 69.0 mg dry weight/liter and 51.6
mg dry weight/liter were attained in Treatment 6 of Stations 32 and
2, respectively (Table 10).
The incremental increase in growth with additional increases in
nitrogen spikes at Station 32 follows a straight line relationship of
Y = 23.34 X + 31.55 with a very high correlation coefficient (r) of
0.98, indicating that Station 32 waters in October were not limiting
to growth at the 1.6 mg nitrogen/liter spike and that further additions
of nitrogen would probably increase the average maximum yield.
The end stations, along with Station 11, had relatively low control
growth and also the lowest concentrations of filtered inorganic nitrogen
(0.02 mg/liter) (Table 6).
A straight-line relationship of Y = 21.33 X + 21.09 was calculated
for the growth of Station 2 treatments. The correlation coefficient (r)
of 0.82 is somewhat lower than the (r) calculated for Station 32.
A comparison was made between the actual growth and the predicted growth
of nitrogen spikes (Table 11). The factor used to calculate predicted growth
was derived from information in Table 12. Table 12 information is based upon
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the growth response of the alga, Dunaliella tertiolecta, to selected
levels of nitrate-nitrogen. As seen in Table 12, Dunaliella varies
somewhat in its response to nitrate-nitrogen additions at the salinity
levels tested (16-35 o/oo), but the overall average yield is 31.8 mg
dry weight per mg of nitrate-nitrogen. Actual yields from Halifax
River impacted stations and Station 11 were considerably beyond maximum
predicted yields (Table 11), so the largest average yield factor given
in Table 12 (70 mg dry weight/mg nitrate nitrogen) was selected to
provide comparative data for these extreme situations. This yield factor
is applicable to salinities in the Halifax River which ranged from 16
o/oo at the Tomoka Basin (Station 32) to 28.5 o/oo at the Ponce de Leon
Inlet (Station 2) (Table 1).
The predicted growth increased progressively by 14 mg dry weight
with each additional 0.2 mg nitrogen spike (Table 11). By adding the
spike yield estimate to the actual yield of a station's control sample,
an estimate for the control growth plus nitrogen spike growth was
determined (Table 11).
The addition of 0.2 mg/liter nitrogen-spikes to impacted stations
and Station 11 water samples increased growth (59.0 - 169.5 mg dry weight/
liter) beyond that predicted (22.3 - 71.9 mg dry weight/liter). The actual
growth was 2.4 to 5.2 times greater than the predicted growth.
It appears that the addition of nitrogen to the impacted stations
and Station 11 stimulates or activates some substance or combination of
substances in sewage which causes the greater growth observed beyond the
growth predicted. The Eutrophication and Lake Restoration Branch National
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Environmental Research Center, Corvallis, Oregon also observed
greater growth than that predicted in research with Dunaliella, but
could not relate it to any particular cause (9).
It is well-known from Liebig's law of the minimum that by comparing
growth per nitrogen spike, a "point" will be reached where growth starts
to level off with increasing spikes. At the nitrogen-spike level where
growth begins leveling off, some other substance becomes limiting to algal
growth. As long as adequate substances are available, an algal population
continues to grow within limits of available light and space. As the supply
of some required substance is used up, the algal population is no longer
able to obtain the substance in amounts needed for further growth.
Based on the algal assay results (Figure 3 and Table 10), the impact
stations and Station 11 nitrogen inputs beyond 0.2 mg/liter do not sub-
stantially increase growth from one spike level to the next indicating
that some other substance becomes limiting to algal growth in excess of
the 0.2 mg nitrogen/liter inputs.
An examination of Figure 3 control (Treatment 2) growth at each
station shows what was expected from our knowledge of the system. Rela-
tively low growth was found near the headwaters of the Halifax River where
there is less population than other areas along the river. Further down-
stream, the growth at Station 9AA reached a maximum of 57.9 mg dry weight/
liter near Daytona Beach (Figure 1), an area where there is known point
waste discharges and less flushing due to causeways traversing the river.
Station 9AA growth was significantly greater than growth from other stations
(Appendix D). Downstream growth decreased at 8AA and attained a minimum of
8.3 mg dry weight/liter at Station 11 near the Ponce de Leon Inlet (Figure 1)
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Growth then increased to 33.8 mg dry weight/liter at end Station 2 near
Smyrna Beach (Figure 1).
A comparison of August and October control growth between stations
(Appendix E) showed significant differences at all stations, except Station
2. The end control stations and Stations 24 and 11 had the greatest growth
during August (Appendix E). Impacted Stations 9AA and 8AA had the greatest
growth during October (Appendix E); this indicates more nitrogen was available
to the algae in October samples at these two impacted stations. The signi-
ficantly greater growth at Station 9AA is not unexpected; filtered total
nitrogen is twice as great in October as in August samples.
The addition of 0.2 mg nitrogen/liter to August and October water samples
caused an increase over control growth at all stations except the Station
2 October sample (Appendix F). October spiked sample growths were signi-
ficantly greater than August spiked sample growths at all stations but
Station 11 (Appendix F).
Chlorophyll samples were only collected during the month of August
and were higher at the impacted stations than the end stations and Station
11 (11) . Levels ranged from 23 yg chlorophyll a/liter to 44 yg chlorophyll
a_/liter at the impacted stations, whereas levels were much reduced at the
other stations ranging from 6 yg chlorophyll a_/liter to 10 yg chlorophyll
a/liter (11).
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DISCUSSION
Nitrogen analyses of the August Halifax River water samples indicated
that the highest level of total nitrogen (0.61 - 0.72 mg/liter) was found in
the impacted area between Stations 24 and 8AA (Table 6). These stations
are located from the Tomoka Basin south to the City of South Daytona Beach
(Figure 1). Lowest levels (0.41 and 0.46 mg/liter) of total nitrogen (Table
6) were found at Stations 11 and 2, near the Ponce de Leon Inlet. Total
nitrogen levels in the Halifax samples were moderate (Table 6) compared
to the 1.0 mg/liter level in over 60% of Florida's salt waters (5).
Phosphorus data of August unfiltered Halifax River water samples (Table
6) indicate that all stations exceed the accepted state level of 0.06
mg/liter total phosphorus (5). Highest levels of total phosphorus were
found at impacted Stations 24, 9AA and 8AA (Table 6). These three stations
are adjacent to the most populated area of Valusia County, namely, the area
near the cities of Ormond Beach, South Daytona Beach and Daytona Beach (Figure
1). FDER found that STORET records of total phosphorus in the Halifax
River exceeded acceptable state levels of 0.06 mg/liter (Table 13) most
of the time (1)- It found that sampling stations in the Daytona Beach
area average 0.28 mg/liter of total phosphorus (1).
The lowest level of total phosphorus (0.07 mg/liter) in the August Halifax
River samples was found in Station 2. Station 2 levels are considerably
below the 0.1 mg phosphorus/liter found in over 60% of Florida's marine
waters (5)- Station 2 is flushed daily by the tide and is a considerable
distance from the densely populated centers of the Halifax River.
Domestic sewage and rural runoff have been cited as the major sources
of excess nitrogen and phosphorus in aquatic systems (5). Sewage effluent
from cities adjacent to the Halifax River contain considerable nutrients (1).
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According to FDER, water samples collected in the vicinity of Holly
Hill, Daytona Beach, and Port Orange have all exceeded acceptable
nutrient levels for phosphates and organic and inorganic nitrogen
(1).
Finding comparatively high nitrogen and phosphorus concentrations in the
study area is not surprising because sewage plants between Ormond Beach and Port
Orange discharge about 15.5 million gallons of treated waste per day (1).
The presence of causeways in this stretch of the River contribute to long
residence times, thus allowing for moire accumulation of treated sewage between
these two communities.
Marine phytoplankton have a varied demand for nitrogen and phosphorus.
Evidence of algal nitrogen and phosphorus requirements is provided by
correlations between algal growth and water chemistry, estimates of the
amounts of nutrients removed by the algal crop, and a comparison of the
chemical composition of naturally growing cells with that known to develop
in cultures under nutrient limiting and enriched conditions (8). The ratio
in which these two elements are utilized is comparable to their atomic
ratio in phytoplankton. A N:P ratio implies that a certain number of
nitrogen atoms are necessary for growth for each phosphorus atom utilized.
N:P ratios of 5:1 - 15:1 are most commonly found in algae (3). Harvey
obtained an average ratio of 16.7:1 in net phytoplankton obtained from
Long Island Sound (4).
According to a review by FDER (5), the average N:P ratio necessary for
optimum growth is about 10:1. This is roughly the point at which nitrogen
and phosphorus may be assumed to be in balance for the needs of a particular
alga. The N:P ratio is useful in preliminary assessment of algal growth
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limitations in natural waters. Water containing N:P ratios less than
10 may be considered nitrogen limited, while those with N:P ratios
greater than 10 may be considered phosphorus limited (5). The N:P
ratio of total nitrogen to total phosphorus for the Halifax River
August chemical analyses (Tables 6 and 7) reflects the large amount
of phosphorus in the Halifax River. The impacted stations had N:P
ratios of 2.1:1 - 2.5:1 for unfiltered samples (Table 6). Stations 32,
11, and 2 located at the headwaters and near the mouth of the Halifax
River had ratios of 3.7:1 - 5.9:1. About 50% of all Florida salt water
areas have N:P ratios of less than 10:1, possibly reflecting the high
natural level of phosphorus in Florida waters (5).
Using the 10:1 N:P ratio for optimum algal growth (5) with the August
water chemistry data, it is probable that all available nitrogen would be
utilized by phytoplankton before all of the available phosphorus. Studies
conducted in New York Harbor indicate that phytoplankton utilize nitrogen
in all forms almost as fast as it is available, while there is surplus of
phosphorus remaining (3). Ammonia is the first nitrogen form utilized
by Dunaliella, but nitrate-nitrite nitrogen will produce the same yield
in Dunaliella cultures if given enough time (2). Some phytoplankton
can utilize equally well nitrate, or ammonia (6). In fact, some algae
can utilize simple organic compounds such as urea and amino acids (5) (6).
The chemical data and N:P ratios of the Halifax River indicate that
nitrogen is the limiting nutrient in the Halifax River study area (Tables
6 and 7). Nitrogen is usually the limiting factor in marine coastal waters
(3), and it is not unusual to find twice the amount of phosphate in marine
waters that algae need (3).
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In addition to the above information, August algal assays further
confirmed that nitrogen was the limiting nutrient in Halifax River water
samples (Tables 4 and 5, Figure 2). Nitrogen limitation in this particular
study implies that more phosphorus was available for growth than nitrogen
and, therefore, the test alga was more responsive or sensitive to additional
nitrogen inputs than additional phosphorus inputs. Actually, there were
sufficient amounts of both nutrients (Table 6) including nitrogen to cause
considerable growth at all sampled stations. That is what happened (Figures
2 and 3), and what was expected in light of the chemistry data (Table 6)
and the fact that Dunaliella can respond to nitrate-nitrogen levels as low
as 0.01 mg/liter and phosphorus levels as low as 0.0025 mg/liter. Also,
EDTA additions showed that metal toxicity had no or very little affect on
growth (Tables 4 and 5, Figure 2).
August algal assays showed that at all stations nitrogen-spiked waters
had greater growth than control flasks, or flasks spiked with phosphorus
or EDTA (Table 4 and Figure 2).
The growth in nitrogen-spiked samples was significantly greater than
control and/or phosphorus-spiked samples at all stations except at Station
2. Sensitivity to increased nitrogen inputs were pronounced at impacted
Stations 9AA and 8AA (Tables 4 and 7, Figure 2) where large amounts of
treated sewage enter the Halifax River (1).
The chemical data, low N:P ratios, and greater algal growth, from
nitrogen spikes led to the determination that nitrogen was the chief
limiting nutrient in the Halifax River, and that it will be the primary
nutrient causing potential nuisance problems with further additions of
sewage. That is why the October algal assays were directed to a study of
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Dunaliella's growth at equivalent or greater nitrogen additions than
those used in the August algal assays. The objective of studying incre-
mental nitrogen inputs into Halifax River water samples is to determine
at what nitrogen concentration other nutrients may become limiting, to
relate growth to potential nuisance blooms, and for input to models for
assessing what effects further nitrogen increases will have on various
areas of the Halifax River.
Greatest growth of Dunaliella to the varied nitrate spikes (Table 8,
Figure 3) in October occurred in samples collected near the most densely
populated areas (1). These areas included Stations 24, 9AA, and 8AA
located between Ormond Beach and Port Orange (Figure 1). The section
of the Halifax River where these three stations are located receives large
amounts of sewage from treatment plants. Further downstream at Station 11,
growth decreases somewhat but it was still relatively high (Table 8,
Figure 3). At all four of these stations (24, 9AA, 8AA, and 11) there was
a significant increase in growth with the addition of 0.2 mg nitrogen/liter
(Table 9, Figure 3). Successive additions of nitrogen did not increase
growth substantially beyond that obtained from 0.2 mg nitrogen/liter. Growth
was not as great at Station 11 but this may be partially due to tidal
flushing reducing amounts of growth substances to levels lower than what
they were at the 3 impacted stations. The high yields at the impacted
stations and Station 11 were beyond what was predicted (Table 11) for
increased nitrogen spikes. This increased yield over that predicted is
not unusual; researchers in EPA (9) have also noted similar results
near sewage discharges. They could not explain why actual yield was
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beyond predicted yield. Possibly other simple organic nitrogen
compounds like urea and amino acids (5) (6) were available for growth.
Growth at the two end stations was generally progressively greater
with increasing additions of nitrate-nitrogen and follow a straight-line
relationship. This relationship indicates that nitrogen was still limiting
at these two stations. Station 32's growth closely agreed with the pre-
dicted yield for the 0.2 mg nitrogen/liter spike and Station 2's yield
was always less than the predicted yield (Table 11).
During both algal assay tests, the data indicate that the algae are
sensitive to nitrogen inputs in both the summer and autumn. The
greatest response occurs at those stations located near large sewage
treatment plants and in waters with longer retention times. The algal
assay data indicate that these areas should not receive increased nitrogen
inputs, or serious problems could develop. In fact, growth in the
control sample at all stations (Tables 4 and 8) indicate potential bloom
problems exist at present unless grazing and/or tidal flushing decrease
algal concentrations in the Halifax River. More severe bloom problems
could result with additional nitrogen inputs.
The amount of dry weight biomass yield from the Dunaliella algal assays
can be converted to cells/m&. About 10,000 cells/m^ of Dunaliella equate to
three mg dry weight per liter (2).
The greatest yield of Dunaliella grown in marine algal media (Treatment
1) was 500 mg dry weight/liter which is equivalent to A,168,355 cells/m£
with an average MCV of 139. This yield level is much greater than that
found in nutrient-spiked Halifax River waters where the greatest yield was
191.6 mg dry weight/liter. The cell count that produced 191.6 mg dry weight/
20
-------
liter was 1,246,168 cells/nU with a MCV of 180; therefore, the yield
level of biomass and number of cells in the Halifax River water treatments
indicate there was no crowding of cells and no carbon limitation.
The October control samples from the Halifax River contained sufficient
nutrients to support from 27,000 Dunaliella cells per m£ at Station 11
to 193,000 cells per mi at impacted Station 9AA. An excess of 15,000
cells/mA in a lake is considered to be eutrophic (7)
The concentration of 15,000 cells/m.^ is not necessarily unacceptable,
but an examination of Table 10 shows that at the impacted stations, 24,
9AA and 8AA, potential nuisance problems could exist with the addition
of only 0.2 mg nitrogen. Calculated concentrations of algal cells ranging
from 5.12 x 105/m& to 5.65 x 105/m& are not desirable.
21
-------
LITERATURE CITED
1. Memorandum from Richard Harvey, EPA to Dr. Tim Stuart, Florida
Department of Environmental Regulation. 1976. Water-Planning-
Wasteload Allocations for Halifax River Discharges.
2. EPA. 1974. Marine Algal Assay: Bottle Test. Environmental
Protection Agency. U. S. Govt. Printing Office. Corvallis,
Oregon.
3. Riley, J. 1971. Nitrogen, Phosphorus in the Coastal Marine Environ-
ment. Science. p. 171.
4. Harvey, H. 1940. Nitrogen and Phosphorus Required for the Growth
of Phytoplankton. Jour. Marine Biol. Assoc. of United Kingdom.
24:115-123.
5. Anonymous. 1976. A Review of the Need for Nutrient Regulation in
Florida. Bureau of Water Quality. Dept. of Environmental Regulation.
pp. 1-27.
6. Anonymous. 1971. Eutrophication in Coastal Waters: Nitrogen as
a Controlling Factor. Environmental Protection Agency. Project
16010 EHC.
7- Anonymous. 1975. Model State Water Monitoring Program. U. S. EPA.
440/9-74-002. Hazardous Materials Office.
8. Lewin, R. 1962. Physiology and Biochemistry of Algae. Academic
Press. New York. 929 pages.
9. Specht. D. 1977- EPA. Personal communication.
10. Ostle, B. 1963. Statistics in Research. Iowa State University
Press. 585 p.
11. Briley-Wild and Associates and Russel & Axion. 1977. Preliminary
Report, Coastal Valusia County Water Quality Sampling and Analytical
Program.
22
-------
FIGURE 1
HALIFAX RIVER
VALUSIA COUNTY, FLORIDA
Tomoka Basin
Tomoka Rive
ORMOND
BEACH
HOLLY
HILL
DAYTONA
BEACH
24
9AA
SOUTH
DAYTONA
PORT
ORANGE
AT L A NTIC
OCEAN
Bridge
© Station
(Map not drawn to scale)
PONCE de LEON INLET
23
-------
PAGE NOT
AVAILABLE
DIGITALLY
-------
PAGE NOT
AVAILABLE
DIGITALLY
-------
TABLE 1
Halifax River Salinity, August, I9T6
STATION
32 24 9AA 8AA 11
16 o/do 18.5 o/oo 18.5 o/oo 18.5 o/oo—r- 27.5 0/oo 28.5 o/oo
Port Orange Causeway
26
-------
TABLE 2
Nutrient and EDTA Additions to Halifax River.Water Samples, August, 1976
Treatments:
1. Reference medium.
2. Sample water (control).
3. Sample water plus 0.02 mg phosphorus per liter.
4. Sample water plus 0.20 mg nitrogen per liter.
5. Sample water, plus 0.02 mg phosphorus and 0.20 mg nitrogen per liter.
6. Sample water plus 1 mg EDTA per liter.
7. Sample water plus 1 mg EDTA and 0.02 mg phosphorus per liter.
8. Sample water plus 1 mg EDTA and 0.2 mg nitrogen per liter.
9. Sample water plus 1 mg EDTA, 0.02 mg phosphorus, and 0.20 mg
nitrogen per liter.
27
-------
TABLE 3
Nitrogen Additions to Halifax River Water Samples, October, 1976
Treatments:
1. Reference algal assay medium.
2. Sample water (control).
3. Sample water plus 0.2 mg nitrogen per liter.
4. Sample water plus 0.4 mg nitrogen per liter.
5. Sample water plus 0.8 mg nitrogen per liter.
6. Sample water plus 1.6 mg nitrogen per liter.
28
-------
TABLE 4
Average Maximum Yields (mg dry weight/liter), Halifax River Water Samples,
August, 1976
Stations
Treatment
2
3
4
5
6
7
8
9
Spike
Control Halifax
River Water
.02 mg P/l
.2 mg N/l
.02 mg P/l
.2 mg N/l
1.0 mg EDTA/1
.02 mg P/l
1.0 mg EDTA/1
.2 mg N/l
1.0 mg EDTA/1
.2 mg N/l
.02 mg P/l
1.0 mg EDTA
32
Mean
41.5
40.1
75.1
73.7
43.4
60.0
67.6
71.3
S.D.*
2.2
3.0
13.7
5.9
11.6
13.6
4.4
9.0
2
Mean
29.9
30.2
58.1
84.5
30.7
32.4
62.2
48.1
4
S.D.
1.7
3.5
18.4
22.2
8.7
20.2
12.5
28.8
9;
Mean
44.5
48.4
94.8
101.1
61.5
46.8
67.8
93.4
\A
S.D.
3.1
1.3
42.9
37.6
16.9
7.4
8.2
2.7
8Aj
Mean
21.4
25.7
62.3
52.8
23.0
23.2
70.6
64.6
{
S.D.
1.5
3.2
11.0
6.2
14.9
1.0
16.6
27.8
i:
Mean
28.3
25.2
62.8
55.0
16.0
28.7
68.3
59.6
L
S.D.
4.0
2.4
6.8
4.7
10.1
3.2
3.7
5.3
2
Mean
49.8
30.9
60.6
43.7
55.3
51.4
75.4
60.2
S.D.
16.0
19.0
4.1
2.1
14.6
14.1
15.2
16.5
S.D. = Standard Deviation
-------
TABLE 5
Comparison of Average Maximum Yield Within Stations
August, 1976
Stations
Treatment
C(2)* vs N(4)
P(3) vs
C(2) vs
N(4) vs
C(2) vs
P(3) vs
N(4) vs
N+P (5)
N(4) vs
N(4)
P(3)
P+N(5)
EDTA(6)
P+EDTA(7)
N4-EDTA(8)
vs N+P+EDTA(9)
N+P+EDTA(9)
32
**
S.05
S.05
S.15
NS
NS
s.io
NS
NS
NS
24
S.15
S.10
NS
S.15
NS
NS
NS
NS
NS
9AA
S.15
S.15
S.15
NS
S.15
NS
NS
NS
NS
8AA
S.05
S.05
S.15
NS
NS
NS
NS
NS
NS
11
S.05
S.05
NS
NS
NS
NS
NS
NS
NS
2
Aft*
NS
S.10
NS
NS
NS
NS
S.15
S.15
NS
ftft*
Treatment control or nutrient spike addition. Treatment number is in
parenthesis.
Significant at a level in subscript when compared to cumulative t-
distribution in Ostle, Appendix 5 p. 528.
NS - not significant at a level.
-------
TABLE 6
Chemical Analyses of Halifax River Water Samples in mg/liter
Station
32
24
9AA
8AA
11
2
U6
.02
.01
.04
.01
.02
.01
N02+NOS-N
\t
F7
.02
.02
.04
.03
.02
.01
cP
U F
01 .01
25 .25
25 .23
05 .06
01 .01
01 .01
A
U
.06
.03
.14
.05
.33
.35
NH^-N
F
.07
.03
.14
.07
.24
.14
Total Inorganic-N
0
U
.01
.01
.10
.05
.12
.17
F U
.01 .08
.01 .03
.13 .18
.01 .06
.01 .35
.01 .36
A
F
.09
.05
.18
.10
.26
.15
0
U
.02
.26
.35
.10
.13
.18
F
.02
.26
.36
.07
.02
.02
TKN2
A
U F
.54 .54
.60 .36
.68 .60
.62 .42
.44 .32
.40 .20
Station
32
24
9AA
8AA
11
2
TKN
0
U F
.50 .47
.50 .30
.47 .40
.42 .30
.20 .10
.20 .10
3
Organic Nitrogen
A 0
U F U F
.48 .49 .49 .46
.57 .33 .49 .29
.54 .46 .37 .25
.57 .35 .37 .29
.11 .08 .08 .09
.05 .06 .03 .09
Total N
A 0
U F U F
.56 .56 .51 .48
.61 .38 .75 .55
.72 .30 .72 .63
.63 .45 .49 .36
.46 .34 .21 .11
.41 .21 .21 .11
Total P
A
U F
.15 .10
.24 .20
.34 .26
.30 .21
.12 .10
.07 .04
Ratio
Total N.: . Total P
A
U F
3.7 5.6
2.5 1.9
2.1 1.2
2.1 2.1
3.8 3.4
5.9 5.3
Total inorganic nitrogen = NO 2 +
2TKN = Total Kjeldahl nitrogen
^Organic N = TKN minus NH3~N
A = August
- N plus NH3 - N
50 = October
= Unfiltered
7F = Filtered through 0.45y millipore filter
-------
TABLE 7
Comparison of Average Maximum Yields and Chemical Analyses
•of Halifax River Water Samples, August, T976
Treatments
Chemical Analyses in mg/liter
Stations
32
24
9AA
8AA
11
2
0.2 mg nitrogen
Control (2) * per liter (4) Difference
41.5
29.9
44.5
21.4
28.3
49.8
75.1
58.1
94.8
62.3
62.8
60.6
+ 33.6
+28.2
+ 50.3
+40.9
+ 34.5
+10.8
Filtered Filtered Filtered
Total P Total N N:P
.10
.20
.26
.21
.10
.04
.56
.38
.30
.45
.34
.21
5.6
1.9
1.2
2.1
3.4
5.3
^Treatment number in parantheses.
32
-------
TABLE 8
Average Maximum Yields (mg dry weight/liter)
Halifax River Water Samples, October, 1976
Treatment
Halifax River
Water Only (2)
0.2 mg N/l (3)
0.4 mg N/l (4)
0.8 mg N/l (5)
1.6 mg N/l (6)
3
Mean
28.6
36.0
—
49.0
69.0
2
S.D.*
7.7
21.2
—
13.4
8.8
2i
Mean
18.3
166.2
166.7
191.6
189.7
t
S.D.
5.0
13.7
16.4
20.6
25.3
Halifax River Stations
9A
Mean
57.9
169.5
149.8
—
163.9
A
S.D.
2.6
15.0
25.0
—
26.7
8£
Mean
35.6
153.4
147.7
—
174.2
iA
S.D.
0.6
15.6
8.8
—
15.4
1
Mean
8.3
59.0
62.4
75.1
76.8
1
S.D.
2.1
14.1
8.3
6.3
14.5
2
Mean
33.8
28.8
42.4
—
51.6
S.D.
4.2
10.4
10.5
—
13.2
S. D. = Standard Deviation
( ) Treatment Number
-------
TABLE 9
Comparison of Average Maximum Yields
Within Stations, October, 1976
Stations
Treatment 32 24 9AA 8AA 11
2 (control) vs 3(.2)* NS*** S<
2 (control) vs 4 (.4) — S
.05 S.05
j-\ c O /-i r
S.05
A q
S.05 NS
S.n.s NS
2 (control) vs 5(.8)* S<10 S>05 ~ — S%05
2 (control) vs 6(1.6)* S<05 S>05 S-05 S-05 S>05
mg nitrogen spike per liter in parentheses.
'significant at a level in subscript when compared to cumulative t-
distribution in Ostle, Appendix 5, p. 528.
not significant at the a level.
34
-------
TABLE 10
Average Maximum Yield of Dunaliella
(mg dry wt./liter)
in October, 1976 Nitrogen spiked Halifax River Water Samples
Stations Treatments in mg N/liter Differences
2 3456
0 .2 .4 .8 1.6 .2-0 .4-.2 .8-.4 1.6-.8
32 28.6 36.0 -- 49.0 69.0 7.4 — — 20.0
24 18.3 166.2 166.7 191.6 189.7 147.9 0.5 24.9 -1.90
9AA 57.9 169.5 149.8 — 163.9 111.6 -19.7
8AA 35.6 153.4 147.7 — 174.2 117.8 -5.7
11 8.3 59.0 62.4 75.1 78.8 50.7 3.4 12.7 3.7
2 33.8 28.8 42.4 — 51.6 20.3 13.6
35
-------
TABLE 11
Predicted* and Actual Average Maximum Yields, (mg dry weight/liter)
for Dunaliella Ocotber Halifax Control and Added Nitrate Spikes
Stations
Treatment 32 24 9AA 8AA 11 2
Halifax River Control
Actual Yield 28.6 18.3 57.9 35.6 8.3 33.8
+.2 mg NaN03/l (predicted yield 14 mg/1)
Actual Yield 40.3 166.2 169.5 153.7 59.0 28.8
Control Yield +
14 mg (predicted) 42.6 32.3 71.9 49.6 22.3 57.8
+.4 mg NaN03/l (predicted yield 28 mg/1)
Actual Yield 166.7 149.8 147.7 62.4 42.4
Control Yield +
28 mg (predicted) 46.3 85.9 63.6 36.3 61.8
+.8 mg NaNCh/1 (predicted yield 56 mg/1)
Actual Yield 49.0 191.6 75.1
Control Yield +
.56 mg (predicted) 84.6 74.3 64.3
+1.6 mg NaN03/l (predicted yield 112 mg/1)
Actual Yield 69.0 189.7 163.9 174.2 76.8 51.6
Control Yield +
112 mg (predicted) 140.6 130.3 169.9 147.6 120.3 145.8
^Predicted yield based on Marine Algal Assay Procedure Bottle Test
36
-------
TABLE 12
Dunaliella dry weight (ing) produced per unit of nitrate-N
at day 14 in defined medium*
Salinity Dry weight (mg) produced per mg of nitrate-N
Mean Range
16 o/oo 30.8 -.01 to 70
20 o/^o 33.1 -.01 to 71
35 o/oo 31.5 -.01 to 68
Ave. 31.8 -.01 to 70
Vrom Marine Algal Assay Procedure Bottle Test (2).
37
-------
TABLE 13
Acceptable and Actual Levels of Nutrients for Halifax River
tug/liter (FDER Criteria)
Nutrient
Organic nitrogen
Ammonia (as N)
Nitrite-Nitrate (as N)
Total Kjeldahl
nitrogen
Phosphate (as P)
Acceptable Level
Halifax River
System
0.70
0.20
0.50
0.90
0.06
Tomoka
River
0.62-1.04
0.02-0.21
0.08-0.18
0.72
0.04-0.27
Actual Levels
Daytona
Beach
0.58-1.01
0.04-0.25
0.16-0.27
1.22
0.11-0.46
Ponce de Leon
Inlet
0.36
0.09
0.05
0.35
0.04
38
-------
LEGEND FOR APPENDICES
1. Treatment = controls or chemical spikes
2. Rep = Replication number
3. Mean = arithmetic mean
4. S = standard deviation
5. d.f. = degrees of freedom
6. t calc. = calculated t
7. two-way t distribution (from Ostle Appendix 5, page 528)
at ct=.15, ct=.10, a=.05
8. Significance - * = significant at a=.15
** = significant at a=.10
*** = significant at a=.05
9. Maximum Duniella terticola yield In mg dry weight per liter.
-------
APPENDIX A
Comparison (t-test) of treatments within stations
Halifax River water samples, August, 1976
-------
APPENDIX A
Station
32
Treatment
2
4
3
4
2
7
3
7
2.
3 -
2
6
2
8
3
8
4
8
5
9
4
5
4
9
Rep2 1
42. 89
41.6
42.8
41.6
42.8
41.6
42.8
31.7
42.8
72.0
41.6
72.0
65.4
72.0
76.1
78.3
65.4
•76.1
65.4
78.3
Rep 2
39.0
65.4
36.6
65.4
39.0
69.7
36.6
69.7
39.0
36.6
39.0
55.0
39.0
67.5
36.6
67.5
84.8
67.5
66.9
74.4
84.8
66.9
84.8
74.4
Rep 3
42.7
84.8
42.0
84.8
42.7
50.4
42.0
50.4
42.7
42.0
42.7
43.4
42.7
63.3
42.0
63.3
63.3
78.0
61.1
78.0
61.1
Mean-*
41.5
75.1
40.1
75.1
41.5
60.0
40.1
60.0
41.5
40.1
41.5
43.4
41.5
67.6
40.1
67.6
75.1
67.6
73.7
71.3
75.1
73.7
75.1
71.3
S4
2.2
13.7
3.0
13.7
2.2
13.6
3.0
13.6
2.2
3.0
2.2
11.6
2.2
4.4
3.0
4.4
13.7
4.4
5.9
9.0
13.7
5.9
13.7
9.0
d.f.5
3
3'
3
3
4
4
4
4
3
4
3
3
t. calc.6
4.535
4.628
2.516
2.652
.669
.272
9 . 301
9.015
.946
.385
.169
.388
.157
1.638
1.638
1.638
1.638
1.533
1.533
1.533
1.533
1.638
1.533
1.638
1.638
.107
2.353
2.353
2.132
2.132
2.132
2.132
2.132
2.132
2.353
2.132
2.353
2.353
.057
3.182
3.182
2.776
2.776
2.776
2.776
2.776
2.776
3.182
2.776
3.182
3.182
Significance
**&
**>v
**
**
***
>v & *
-------
APPENDIX A (Continued)
dtion
24
Treatment
2
4
2
8
2
9
2
. 3
2
6
2
5
3
8
3 '
4 .
3
7
4
8
4
5
4
9
Rep2 1
31.1
62.3
31.1
76.6
31.1
62.9
31.1
31.8
31.1
39.2
31.1
64.8
31.8
76.6
31.8
62.3
31.8
55.2
62.3
76.6
62.3
. 64.8
62.3
62.9
Rep 2
28.7
38.0
28.7
54.2
28.7
15.0
28.7
32.6
28.7
21.8
28.7
108.5
32.6
54.2
32.6
38.0
32.6
25.5
38.0
54.2
38.0
108.5
38.0
15.0
Rep 3
74.1
55.7
66.5
26.2
31.0
80.3
26.2
55.7
26.2
74.1
26.2
16.6
74.1
55.7
74.1
80.3
74.1
66.5
•3
Mean
29.9
58.1
29.9
62.2
29.9
48.1
29.9
30.2
29.9
30.7
29.9
84.5
30.2
62.2
30.2
58.1
30.2
32.4
58.1
62.2
58.1
84.5
58.1
48.1
S*
1.7
18.4
1.7
12.5
1.7
28.8
1.7
3.5
1.7
8.7
1.7
22.1
3.5
12.5
3.5
18.4
3.5
20.2
18.4
12.5
18.4
22.2
18.4
28.8
d.f.5
3
3
3
3
3
3
4
4
4
4
4
4
t. calc.6
2.053
3.441
.836
.109
.117
3.303
4.259
2.582
.188
.313
1.587
.507
.157
1.638
1.638
1.638
1.638
1.638
1.683
1.533
1.533
1.533
1.533
1.533
1.533
.107
2.353
2.353
2.353
2.353
2.353
2.353
2.132
2.132
2.132
2.132
2.132
2.132
.057
3.182
3.182
3.182
3.182
3.182
3.182
2.776
2.776
2.776
2.776
2.776
2.776
Significance
*
ft**
ftftft
ftftft
ftft
ft
-------
APPENDIX A (Continued)
Station
24
9AA
Treatment1
5
9
6
8
7
8
6
9
7
9 '
2
6
2
3
2
4
2
5
2
8
8
9
4
9
Rep2 1
64.8
62.9
39.2
76.6
55.2
76.6
39.2
62.9
55.2
62.9
41.9
54.4
41.9
47.0
41.9
61.8
41.9
74.5
41.9
70.4
70.4
' 90.6
61.8
90.6
Rep 2
108.5
15.0
21.8
54.2
25.5
54.2
21.8
15.0
25.5
15.0
43.8
49.2
43.8
48.6
43.8
143.3
43.8
127.7
43.8
74.5
74.5
93.6
143.3
93.6
Rep 3
66.5
31.0
55.7
16.6
55.7
31.0
66.5
16.6
66.5
47.9
80.8
47.9
49.6
47.9
79.3
47.9
47.9
58.6
58.6
96.0
79.3
96.0
0
MeanJ
86.6
48.1
30.7
62.2
32.4
62.2
30.7
48.1
32.4
48.1
44.5
61.5
44.5
48.4
44.5
94.8
44.5
101.1
44.5
67.8
67.8
93.4
94.8
93.4
S^
30.9
28.8
8.7
12.5
20.2
12.5
8.7
28.8
20.2
28.8
3.1
16.9
3.1
1.3
3.1
42.9
3.1
37.6
3.1
8.3
8.3
2.7
42.9
2.7
d.f.5
3
4
4
4
4
4
4
4
3
4
4
4
t. calc.6
1.431
3.577
2.165
1.007
.774
1.703
2.008
2.024
2.834
4.582
5.097
.056
.157
1.638
•1.533
•1.533
1.533
1.533
1.533
1.533
1.533
1.638
•1.533
1.533
1.533
.107
2.353
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.353
2.132
2.132
2.132
.057
3.182
2.776
2.776
2.776
2.776
2.776
2.776
2.776
3.182
2.776
2.776
2.776
Significance
&&*
**
*
A
#
**
***
***
-------
APPENDIX A (Continued)
Station
9AA
Treatment
6
9
3
8
3
5
4
.5
4
8' .
5
8
2
9
3
9
7
9
5
9
3
7
Rep2 1
54.4
90.6
47.0
70.4
47.0
74.5
61.8
74.5
61.8
70.4
74.5
70.4
41.9
90.6
47.0
90.6
42.4
90-. 6
74.5
90.6
47.0
.42.2
Rep 2
49.2
93.6
48.6
74.5
48.6
127.7
143.3
127.7
143.3
74.5
127.7
74.5
43.8
93.6
48.6
93.6
42.6
93.6
L27.7
93.6
48.6
42.6
Rep 3
80.8
96.0
49.6
58.6
49.6
79.3
79.3
58.6
58.6
47.9
96.0
49.6
96.0
55.3
96.0
96.0
49.6
55.3
0
MeanJ
61.5
93.4
48.4
67.8
48.4
101.1
94.8
101.1
94.8
67.8
101.1
67.8
44.5
93.4
48.4
93.4
46.8
93.4
101.1
93.4
48.4
46.7
S4
16.9
2.7
1.3
8.3
1.3
37.6
42.9
37.6
42.9
8.3
37.6
8.3
3.1
2.7
1.3
2.7
7.4
2.7
37.6
2.7
1.3
7.4
d.f.5
4
4
3
3
4
3
4
4
4
3
4
t. calc.6
3.233
4.027
2.654
.167
1.069
1.602
20.69'7
25.923
10.262
.386
.389
.157
1.533
1.533
1.638
1.638
1.533
1.638
1.533
1.533
•1.533
1.638
1.533
.107
2.132
2.132
2.353
2.353
2.132
2.353
2.132
2.132
2.132
2.353
2.132
.057
2.776
2.776
3.182
3.182
2.776
3.182
2.776
2.776
2.776
3.182
2.776
Significance
***
***
**
***
5V **
***
-------
APPENDIX A (Continued)
tat ion
9AA
8M
Treatment
3
4
2
3
2
9
2
, 4
3
4
2
6
2
5
3
5
4
7
5
7
7
8
Rep2 1 !
47.0
61.8
21.7
29.1
21.7
43.8
21.7
49.8
29.1
49.8
21.7
8.3
21.7
57.2
29.1
57.2
49.8
22.1
57.2
22.1
22.1
.89.7
Rep 2
48.6
143.3
22.7
25.1
22.7
53.8
22.7
70.8
25.1
70.8
22.7
22.6
22.7
48.5
25.1
48.5
70.8
24.0
48.5
24.0
24.0
60.1
Rep 3
49.6
79.3
19.8
22.8
19.8
96.1
19.8
66.3
22.8
66.3
19.8
38.1
19.8
22.8
66.3
23.6
23.6
23.6
62.0
•3
Mean
48.4
94.8
21.4
25.7
21.4
64.6
21.4
62.3
25.7
62.3
21.4
23.0
21.4
52.8
25.7
52.8
62.3
23.2
52.8
23.2
23.2
70.6
S*
1.3
42.9
1.5
3.2
1.5
27.8
1.5
11.1
3.2
11.1
1.5
14.9
1.5
6.2
3.2
6.2
11.1
1.0
6.2
1.0
1.0
16.6
d.f.5
4
4
4
4
4
4
3
3
4
3
4
t. calc.6
1.872
2.104
2.689
6.350
5.514
.185
9.187
6.762
6.094
8.901
4.942
.157
1.533
1.533
1.533
• 1.533
1.533
1.533
• 1.638
1.638
•1.533
1.638
• -1.533
.107
2.132
2.132
2.132
2.132
2.132
2.132
2.353
2.353
2.132
2.353
2.132
.057
2.776
2.776
2.776
2.776
2.776
2.776
3.182
3.182
2.776
3.182
2.776
Significance
A
*
**
*&*
*ft*
>V*A
#**
*5'oV
*#5V
***
-------
Station
8AA
11
Treatment
2
3
2
4
3
7
4
5
4
8
5
9
4
9
2
5
2
8
2
9
3
5
7
9
Rep2 1
21.7
29.1
21.7
49.8
29.1
22.1
49.8
57.2
49.8
89.7
57.2
43.8
49.8
43.8
26.7
58.4
26.7
64.4
26.7
61.3
26.6
58.4
26.2
61.3
Rep 2
22.7
25.1
22.7
70.8
25.1
24.0
70.8
48.5
70.8
60.1
48.5
53.8
70.8
53.8
25.4
51.7
25.4
71.8
25.4
53.7
26.6
51.7
27.5
53.7
A
Rep 3
19.8
22.8
19.8
66.3
22.8
23.6
66.3
66.3
62.0
96.1
66.3
96.1
32.9
32.9
68.8
32.9
63.8
22.5
32.3
63.8
PPENDIX
Mean
21.4
25.7
21.4
62.3
25.7
23.2
62.3
52.8
62.3
70.6
52.8
64.6
62.3
64.6
28.3
55.0
28.3
68.3
28.3
59.6
25.2
55.0
28.7
59.6
A (Cont
s4
1.5
3.2
1.5
11.1
3.2
1.0
11.1
6.2
11.1
16.6
6.2
27.8
11.1
27.8
4.0
4.7
4.0
3.7
4.0
5.3
2.4
4.7
3.2
5.3
inued)
d.f.5
4
4
4
3
4
3
4
3
4
4
3
4
t. calc.
2.104
6.350
1.261
1.067
.721
.559
.131
6.816
12.666
8.189
9.752
8.692
.157
1.533
1.533
1.533
1.638
1.533
1.638
1.533
1.638
1.533
1.533
1.638
1.533
.io7
2.132
2.132
2.132
2.353
2.132
2.353
2.132
2.353
2.132
2.132
2.353
2.132
.057
2.776
2.776
2.776
3.182
2.776
3.182
2.776
3.182
2.776
2.776
3.182
2.776
Significance
*
AA*
AAA
AAA
AAA
AA*
AAA
-------
APPENDIX A (Continued)
Station
11
Treatment1
3
8
3
9
4
6
5
- 6
6
8 -
6
9
4
7
5
7 '
7
8
2
3
2
6
4
9
Rep2 1
26.6
64.4
26.6
61.3
57.9
21.6
58.4
21.6
21.6
64.4
21.6
61.3
57.9
26.2
58.4
26.2
26.2
64.4
26.7
26.6
26.7
- 21.6
57.9
61.3
Rep 2
26.6
71.8
26.6
53.7
67.6
4.4
51,7
4.4
4.4
71.8
4.4
53.7
67.6
27.5
51.7
27.5
27.5
71.8
25.4
26.6
25.4
4.4
67.6
53.7
Rep 3
22.5
68.8
22.5
63.8
22.1
22.1
22.1
68.8
22.1
63.8
32.3
32.3
32.3
68.8
32.9
22.5
32.9
22.1
63.8
MeanJ
25.2
68.3
25.2
59.6
62.8
16.0
55.0
16.0
16.0
68.3
16.0
59.6
62.8
28.7
55.0
28.7
28.7
68.3
28.3
25.2
28.3
16.0
62.8
59.6
S*
2.4
3.7
2.4
5.3
6.9
10.1
4.7
10.1
10.1
3.7
10.1
5.3
6.9
3.2
4.7
3.2
3.2
3.7
4.0
2.4
4.0
10.1
6.9
5.3
d.f.5
4
4
3
3
4
4
3
3
4
4
4
3
t. calc.6
16.924
10.319
5.604
4.929
8.431
6.637
7.860
7.625
13.972
1.153
1.964
.590
.157
1.533
1.533
1.638
1.638
1.533
•1.533
•1.638
1.638
1.533
1.533
1.533
1.638
.107
2.132
2.132
2.353
2.353
2.132
2.132
2.353
2.353
2.132
2.132
2.132
2.353
.057
2.776
2.776
3.182
3.182
2.776
2.776
3.182
3.182
2.776
2.776
2.776
3.182
Significance1
**&
&*&
***
***
***
***
***
***
***
•k
-------
APPENDIX A (Continued)
>tation
11
2
Treatment
2
4
3
4
3
7
4
.8
4.
5 -
5
9
5
8
2 '
6
3
7
4
8
5
9
Rep2 1
26.7
57.9
26.6
57.9
26.6
26.2
57.9
64.4
57.9
58.4
58.4
61.3
58.4
64.4
59.5
65.6
41.8
41.5
56.3
89.8
42.0
-66.2
Rep 2
25.4
67.6
26.6
67.6
26.6
27.5
67.6
71.8
67.6
51.7
51.7
53.7
51.7
71.8
58.7
45.0
42.0
61.4
61.2
76.9
4,6.0
41.5
Rep 3
32.9
22.5
22.5
32.3
68.8
63.8
68.8
31.3
8.9
64.4
59.4
43.0
72.8
Mean3
28.3
62.8
25.2
62.8
25.2
28.7
62.8
68.3
62.8
55.0
55.0
59.6
55.1
68.3
49.8
55.3
30.9
51.4
60.6
75.4
43.7
60.2
S*
4.0
6.9
2.3
6.9
2.4
3.2
6.9
3.7
6.9
4.7
4.7
5.3
4.7
3.7
16.1
14.6
19.0
14.1
4.1
15.3
2.1
16.5
d.f.5
3
3
4
3
2
3
3
3
3
4
4
t. calc.6
7.338
9.261
1.490
1.225
1.306
.978
3.558
.384
1.282
1.615
1.718
.157
1.638
1.638
1.533
1.638
1.886
1.638
1.638
1.638
1.638
1.533
1.533
.107
2.353
2.353
2.132
2.353
2.920
2.353
2.353
2.353
2.353
2.132
2.132
.057
3.182
3.182
2.776
3.182
4.303
3.182
3.182
3.182
3.182
2.776
2.776
Significance
***
***
&&*
*
*
-------
APPENDIX A (Continued)
Station
2
Treatment
2
4
3
4
4
5
6
.8
2.
8 .
2
9
2
3
2
5
4
9
Rep2 1
59.5
56.3
41.8
56.3
56.3
42.0
65.6
89.8
59.5
89.8
59.5
66.2
59.5
41.8
59.5
42.0
56.3
66.2
Rep 2
58.7
61.2
42.0
61.2
61.2
46.0
45.0
76.9
58.7
76.9
58.7
41.5
58.7
42.0
58.7
46.0
61.2
41.5
Rep 3
31.3
64.4
8.9
64.4
64.4
43.0
59.4
31.3
59.4
31.3
72.8
31.3
8.9
31.3
43.0
64.4
72.8
0
Mean0
49.8
60.6
30.9
60.6
60.6
43.7
55.3
75.4
49.8
75.4
49.8
60.2
49.8
30.9
49.8
43.7
60.6
60.2
S4
16.1
4.1
19.1
4.1
4.1
2.1
14.6
15.3
16.1
15.3
16.1
16.5
16.1
19.1
16.1
2.1
4.1
16.5
d.f.5
4
4
4
3
4
4
4
4
4
t. calc.6
1.129
2.643
6.416
1.462
1.996
.777
1.316
.659
.048
.157
1.533
1.533
1.533
1.638
1.533
1.533
1.533
1.533
1.533
.107
2.132
2.132
2.132
2.353
2.132
2.132
2.132
2.132
2.132
.057
2.776
2.776
2.776
3.182
2.776
2.776
2.776
2.776
2.776
Significance
**
***
*
-------
APPENDIX B
Comparison (t-test) of controls (Treatment 2) among stations
Halifax River water samples, August, 1976
-------
APPENDIX B
Station
8AA
11
8AA
2
32
24
32
9AA
32
8AA
32
11
32
2
24
9AA
24
8AA
24
11
24
2
Treatment1
2
2
2
2
2
2
2
- 2
2
2 -
2
2
2
2
2 '
2
2
2
2
2
2
2
Rep2 1
19. 89
25.4
19.8
58.7
39.0
28.7
39.0
43.8
39.0
19.8
39.0
25.4
39.0
58.7
28.7
43.8
28.7
19.8
28.7
25.4
28.7
-58.7
Rep 2
22.7
32.9
22.7
31.3
42.7
31.1
42.7
47,9
42.7
22.7
42.7
32.9
42.7
31.3
31.1
47.9
31.1
22.7
31.1
32.9
31.1
31.3
Rep 3
21.7
26.7
21.7
59.5
42.8
42.8
41.9
42.8
21.7
42.8
26.7
42.8
59.5
41.9
21.7
26.7
59.5
0
Mean
21.4
28.3
21.4
49.8
41.5
29.9
41.5
44.5
41.5
21.4
41.5
28.3
41.5
49.8
29.9
44.5
29.9
21.4
29.9
28.3
29.9
49.8
S4
1.5
4.0
1.5
16.1
2.2
1.7
2.2
3.1
2.2
1.5
2.2
4.0
2.2
16.1
1.7
3.1
1.7
1.5
1.7
4.0
1.7
16.1
d.f.5
4
4
3
4
4
4
4
3
3
3
3
t. calc.6
2.812
3.054
6.285
1.399
13.292
5.006
.890
5.962
6.002
.502
1.661
.157
1.533
•1.533
1.638
1.533
1.533
1.533
1.533
1.638
1.638
1.638
1.638
.107
2.132
2.132
2.353
2.132
2.132
2.132
2.132
2.353
2.353
2.353
2.353
.057
2.776
2.776
3.182
2.776
2.776
2.776
2.776
3.182
3.182
3.182
3.182
Significance
***
***
***
***
*&*
&**
***
*
-------
APPENDIX B (Continued)
Station
9AA
8AA
9AA
11
9AA
2
11
2
Treatment
2
2
2
2
2
2
2
2
Rep2 1
43.8
19.8
43.8
25.4
43.8
48.7
26.7
59.5
Rep 2
47.9
22.7
47.9
32.9
47.9
31.3
25.4
58.7
Rep 3
41.9
21.7
41.9
26.7
41.9
59.5
32.9
31.3
Me an ^
44.5
21.4
44.5
28.3
44.5
49.8
28.3
49.8
s«
3.1
1.5
3.1
4.0
3.1
16.1
4.0
16.1
d.f.5
4
4
4
4
t. calc.6
11.777
5.560
.561
2.250
.157
1.533
•1.533
1.533
1.533
.107
2.132
2.132
2.132
2.132
.057
2.776
2.776
2.776
2.776
Significance
***
***
**
-------
APPENDIX C
Comparison (t-test) of treatments
within stations for October, 1976, Halifax River water samples
-------
APPENDIX C
Station
32
24
9AA
Treatment
2
3
2
6
2
5
5
6
5
3
6
3
2
3
2
4
2
5
2
6
4
5
2
3
Rep2 1
21.0
60.1
21. O9
74.7
21.0
33.9
33.9
74.7
33.9
60.1
74.7
60.1
13.0
170.6
13.0
174.9
13.0
206.2
13.0
167.7
174.9
206.2
60.9
183.0
Rep 2
28.3
27.4
28.3
73.4
28.3
5-9.4
59.4
73.4
59.4
27.4
73.4
27.4
23.0
177.2
23.0
177.3
23.0
177.1
23.0
217.3
177.3
177.1
56.6
172.2
Rep 3
36.4
20.5
36.4
58.9
36.4
53.8
53.8
58.9
53.8
20.5
58.9
20.5
19.0
150.8
19.0
147.8
19.0
19.0
184.0
147.8
56.1
153.3
Mean
28.6
36.0
28.6
69.0
28.6
49.0
49.0
69.0
49.0
36.0
69.0
36.0
18.3
166.2
18.3
166.7
18.3
191.6
18.3
189.7
166.7
191.6
57.9
169.5
s4
7.7
21.2
7.7
8.8
7.7
13.4
13.4
8.8
13.4
21.2
8.8
21.2
5.0
13.7
5.0
16.4
5.0
20.6
5.0
25.3
16.4
20.6
2.6
15.0
d.f.5
4
4
4
4
4
4
4
4
3
4
3
4
t. calc.
.572
5.949
2.293
2.159
.901
2.495
17.503
14.990
15.103
11.512
1.529
L2.668
.157
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.638
1.533
1.638
1.533
.107
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.353
2.132
2.353
2.132
.057
2.776
2.776
2.776
2.776
2.776
2.776
2.776
2.776
3.182
2.776
3.182
2.776
c. ... 8
Significance
***
*&
**
**
***
*&*
5V*-A
***
***
-------
APPENDIX C (Continued)
tation
9AA
8AA
11
Treatment
2
4
2
6
2
3
- 2
4
2 -
6
3
6
3
4
4
6
2
3
2
4
2
5
Rep" 1
60.9
132.1
60.9
192.4
35.0
136.9
35.0
137.7
35.0
191.5
136.9
191.5
136.9
137.7
137.7
191.5
7.4
74.8
7.4
58.8
- 7.4
67.8
Rep 2
56.6
167.4
56.6
160.0
36.0
167.0
36.0
154.0
36.0
162.0
167.0
162.0
167.0
154.0
154.0
162.0
10.7
47.7
10.7
56.6
10.7
78.0
Rep 3
56.1
56.1
139.4
35.9
156.4
35.9
151.5
35.9
169.1
156.4
169.1
156.4
151.5
151.5
169.1
6.7
54.4
6.7
71.9
6.7
79.4
•3
MeanJ
57.9
149.8
57.9
163.9
35.6
153.4
35.6
147.7
35.6
174.2
153.4
174.2
153.4
147.7
147.7
174.2
8.3
59.0
8.3
62.4
8.3
75.1
S4
2.6
25.0
2.6
26.7
.6
15.3
.6
8.8
.6
15.4
15.3
15.4
15.3
8.8
8.8
15.4
2.1
14.1
2.1
8.3
2.1
6.3
d.f.5
3
4
4
4
4
4
4
4
4
4
4
t. calc.6
6.907
6.842
13.355
22.074
15.577
1.658
.560
2.586
6.151
10.981
17.313
.157
1.638
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.533
.107
2.353
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.132
.057
3.182
2.776
2.776
2.776
2.776
2.776
2.776
2.776
2.776
2.776
2.776
Signif icance
AAA
***
***
&*&
A A*
*
A*
AAA
A * *
A**
-------
APPENDIX C (Continued)
11
Treat meat
2
6
4
5
6
5
6
4
6 '
3
5
4
5
3
4
3
2
3
2
4
2
6
Hep2 1
7.4
60.2
58.8
31.3
60.2
67.8
60.2
58.8
60.2
74.8
67.8
58.8
67.8
74.8
58.8
74.8
30.4
37.3
30.4
54.6
~30.4
65.0
Rep 2
10.7
83.2
56.6
85.6
83.2
78.0
83.2
56.6
83.2
47.7
78.0
56.6
78.0
47.7
56.6
47.7
37.2
17.1
37.2
35.9
37.2
38.5
Rep 3
6.7
86.9
71.9
79.4
86.9
79.4
86.9
71.9
86.9
54.4
79.4
71.9
79.4
54.4
71.9
54.4
31.9
36.8
51.3
Mean
8.3
76.8
62.4
65.4
76.8
75.1
76.8
62.4
76.8
59.0
75.1
62.4
75.1
59.0
62.4
59.0
33.8
28.8
33.8
42.4
33.8
51.6
S4
2.1
14.5
8.3
29.7
14.5
6.3
14.5
8.3
14.5
14.1
6.3
8.3
6.3
14.1
8.3
14.1
4.8
10.5
4.8
10.5
4.8
13.3
d.f.5
4
4
4
4
4
4
4
4
3
3
3
t. calc.6
8.113
.168
.186
1.489
1.525
2.100
1.802
.367
.614
1.045
1.745
.15"'
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.533
1.638
1.638
1.638
.107
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.132
2.353
2.353
2.353
.057
2.776
2.776
2.776
2.776
2.776
2.776
2.776
2.776
3.182
3.182
3.182
Signif icjtice
***
&
*
*
-------
APPENDIX C (Continued)
Station
2
Treatment
3
4
3
6
4
6
•
Rep2 1
37.3
54.6
37.3
65.0
54.6
65.0
Rep 2
17.1
35.9
17.1
38.5
35.9
38.5
Rep 3
31.9
36.8
31.9
51.3
36.8
51.3
o
Mean
28.8
42.4
28.8
51.6
42.4
51.6
S4
10.5
10.5
10.5
13.3
10.5
13.3
A f 5
Q . 1 .
4
4
4
t. ealc.6
1.593
2.342
.937
.157
1.533
1.533
1.533
.107
2.132
2.132
2.132
.057
2.776
2.776
2.776
Significance
*
**
-------
APPENDIX D
Comparison (t-test) of controls (Treatment 2) among stations
for October, 1976, Halifax River water samples
-------
APPENDIX D
Station
32
24
32
9AA
32
8AA
32
11
32
2
24
9AA
24
8AA
24
11
24
2
9AA
8AA
9AA
11
Treatment
2
2
2
2
2
2
2
' 2
1
2
2
2
2
2
2
2
2
2
2
2
2
2
Rep2 1
21. O9
13.0
21.0
60.9
21.0
35.0
21.0
7.4
21.0
30.4
13.0
60.9
13.0
35.0
13.0
7.4
13.0
30.4
60.9
35.0
60.9
7.4
Rep 2
28.3
23.0
28.3
56.6
28.3
36.0
28.3
10.7
28.3
37.2
23.0
56.6
23.0
36.0
23.0
10.7
23.0
37.2
56.6
36.0
56.6
10.7
Rep 3
36.4
19.0
36.4
56.1
36.4
35.9
36.4
6.7
36.4
19.0
56.1
19.0
35.9
19.0
6.7
19.0
56.1
35.9
56.1
6.7
Mean3
28.6
18.3
28.6
57.9
28.6
35.6
28.6
8.3
28.6
33.8
18.3
57.9
18.3
35.6
18.3
8.3
18.3
33.8
57.4
35.6
57.9
8.3
^
7.0
5.0
7.7
2.6
7.7
.6
7.7
2.1
7.7
4.8
5.0
2.6
5.0
.6
5.0
2.1
5.0
4.8
2.6
.6
2.6
2.1
d.f.5
4
4
4
4
3
4
4
4
3
4
4
t. calc.6
1.926
6.232
1.584
4.398
.833
12.048
5.918
3.188
3.416
14.285
25.304
.157
1.533
1.533
1.533
1.533
1.638
1.533
1.533
1.533
1.638
1.533
1.533
.107
2.132
2.132
2.132
2.132
2.353
2.132
2.132
2.132
2.353
2.132
2.132
.057
2.776
2.776
2.776
2.776
3.182
2.776
2.776
2.776
3.182
2.776
2.776
Significance
*
***
ft
ftftft
ftftft
ftftft
ftftft
ftftft
ftftft
ftftft
-------
APPENDIX D (Continued)
Station
9AA
2
11
2
8AA
11
8AA
2
i
Treatment
2
2
2
2
2
2
2
'2
,, 2 ,
Rep 1
60.9
30.4
7.4
30.4
35.0
7.4
35.0
30.4
Rep 2
56.6
37.2
10.7
37.2
36.0
10.7
36.0
37.2
Rep 3
56.1
6.7
35.9
6.7
35.9
.„ 3
Mean
57.9
33.8
8.3
33.8
35.6
8.3
35.6
33.8
4
s
2.6
4.8
2.1
4.8
.6
2.1
.6
4.8
j f 5
d.f.
3
3
4
3
i 6
t. calc.
7.502
8.531
21.486
.714
l r 7
. 15 '
1.638
1.638
1.533
1.638
, ,,7
. 10 '
2.353
2.353
2.132
2.353
rvc 7
.05 '
3.182
3.182
2.776
3.182
Significance
&**
***
**&
-------
APPENDIX E
Growth comparison (t-test) of August and October
controls within stations, Halifax River Water Samples
-------
APPENDIX E
Station
32
24
9AA
8AA
11
2
Treatment
A2
02
A2
02
A2
02
A2
02
A2-
02 "
A2
02
Rep2 1
42. 89
21.0
31.1
13.0
41.9
60.9
21.7
35.0
26.7
7.4
59.5
30.4
Rep 2
39.0
28.3
28.7
23.0
43.8
56.6
22.7
36.0
25.4
10.7
58.7
37.2
Rep 3
42.7
36.4
19.0
47.9
56.1
19.8
35.9
32.9
6.7
31.3
Mean
41.5
28.6
29.9
18.3
44.5
57.9
21.4
35.6
28.3
8.3
49.8
33.8
S4
2.2
7.7
1.7
5.0
3.1
2.6
1.5
.6
4.0
2.1
16.1
4.8
d.f.5
4
3
4
4
4
3
t. calc.6
2.799
2.999
5.708
15.675
7.652
1.310
.157
1.533
1.638
1.533
1.533
1.533
[
1.638
I
.107
2.132
2.353
2.132
2.132
2.132
2.353
.057
2.776
3.182
2.776
2.776
2.776
3.182
Significance
***
5V*
&&*
***
&**
-------
APPENDIX F
Growth Comparison (t-test) of August and October
equivalent nitrogen spikes within stations, Halifax River water samples
-------
APPENDIX F
Station
32
24
9AA
8AA
11
2
Treatment
A4
03
A4
03
A4
03
A4
03
A4-
03 '
A4
03
Rep2 1
65. 49
60.1
62.3
170.6
61.8
183.8
49.8
13.. 69
57.9
74.8
56.3
37.3
Rep 2
84.8
27.4
38.0
1-77.2
143.3
172.2
70.8
167.8
67.6
47.7
61.2
17.1
Rep 3
20.5
74.1
150.8
79.3
153.3
66.3
156.4
54.4
64.4
31.9
0
Mean
75.1
36.0
58.1
166.2
94.8
169.5
62.3
153.7
62.8
59.0
60.6
28.8
^
13.7
21.2
18.4
13.7
42.9
15.0
11.1
15.6
6.9
14.1
4.1
10.5
d.f.5
3
4
4
4
3
4
t. calc.6
2.254
8.149
2.846
8.270
.340
4.916
.157
1.638
1.533
1.533
1.533
1.638
!
1.533
.107
2.353
2.132
2.132
2.132
2.353
2.132
.057
3.182
2.776
2.776
2.776
3.182
2.776
Significance
*
***
ft&*
***
***
------- |