ML19308D828

Facility Environ Research Program to Meet Current Federal Requirements.

ML19308D828
Person / Time
Site:	Crystal River
Issue date:	06/11/1973
From:	FLORIDA POWER CORP.
To:
References
NUDOCS 8003191009
Download: ML19308D828 (88)

Text

. _ . _ . _ _ _ _ _

,, l i

l THIS DOCUMENT CONTAINS '

POOR QUAUTY PAGEL _

REBil ATORY DCCET fl.E COPY  !

RETUPJi TO REGU ATORY CMl/J.R.ES  ;

RJ0M 016 i i

)

l l .

.,  ??

)

l J

Florida Power CURPOAAT4ON 8003190 1009

)

Sc - 3 0.t.

C d 0 C !{D J JUN131973m 1

$l!llf.ff:R.1 }

4 $f.$

gpq fil* UY- 4 FLORIDA POWER CORPORATION CRYSTAL RIVER ENVIRONMENTAL RESEARCH PROGR.Of TO MEET 4 CURRENT FEDERAL REQUIPSIENTS JUNE 11 1973 r.rteirst e;ue sua

4t-0 I

S

't 4

mm 9

1 e

.O l

L 3f/f \

Table of Cortents Page I. Introduction 1 II. Project Management 4 III. Statistical Control 7 IV._ Hodelling for Understanding 8 V. Program Description 19 A. Intake Area 19 B. Intake Canal 25 C. Discharge Area 30 D. Marshgrass Study 37 E. Other Areas 38 VI. Exceptions to the Program Proposed by the Federal Agencies ,

A. Plankton Program 42 B. Fish Impingement Program 48 VII. Enclosures A. Interagency Proposes Program 50 B. University of Florida evaluation of the interagency's Plankton Program 59 Proposed program to determine fate and C.

condition of plankton (Maturo and Fox) 70 4

s M

s

1. INTRODUCTION Environmental research at the Crystal River site has been underway for a number of years. The objectives of this research have been and continue to be to:

- 1. Establish the environmental impact of the existing fossil Units 1 and 2 as a baseline for predicting and assessing the impact of Unit 3 nuclear plant now under construction.

2. Establish the environmental impact of Unit 3 nuclear plant during its early operation, and

3. Utilize the research results as a basis for decision making to assure environmental protection in the public interest.

The specific research sr ;egy for achieving these objectives has been adjusted from time to time due to a growing knowledge of environmental conditions in the area and the changing emphasis of various interested groups and governmental agencies. This document has been prepared to

, describe the latent adjustment of the research strategy in response to:

1. U.S. Atomic Energy Commission specification and guidance for environmental research to be completed by November 1974 to determine 1

i l

l' the necd, if any, for modifications of the proposed cooling system (current design - once through 17 degress F maximum 6 T) -'

for Unit 3 nuclear plant as stipulated in the letter of May 8,1973 to Florida Power Corporation from Mr. Dan Muller of the AEC and in the AEC Final Environmental Statement for Crystal River Unit 3 dated May 1973.

2. U.S. Environmental Protection Agency specif"i cations and guidance for environmental research by November 1974 to determine ,

the impact of the existing fossil Units 1 and 2, and Unit 3 as stipulated in a meeting with EPA and other federal agencies at Florida Power Corporation on May 10, 1973.

• In both cases of AEC and EPA research requirements above, coordination between AEC and EPA with other federal agencies has been accomplished and thus represents the consensus specification and guidance of the federal government. The other federal agencies include the Department of the Interior and National Oceanographic and Atmospheric Administration. The required program allows a single research effort that optimizes coordination,

. administration and economy.

In response to these requirements, Florida Power Corporation has moved to

' amend its already comprehensive research effort to incorporate, where practical, these new areas of concern. These include expanded definition of the benthic community in the discharge area, the plankton distribution in both the intake and discharge area and the characteristics of the 2

y

1

~

population of the intake canal. Minor program adjustments in other areas have also been incorporated. . In addition, the programs already underway -

will be continued until their successful completion.

Based on extensive consultations with environmental scientists in the State of Florida, it is our opinion that the program described herein is comprehensive and adequate to define che environmental characteristics of the marine region in the vicinity of the Crystal River site. The means have also been incorporated to interpret this wide base of data as it becomes

-available to: insure a meaningful decision making process. The completion of the program should be realized in the late summer of 1974. This is well within the time frame specified by the AEC. -

a 0

8 m

G m

m D

E s

3 g

0 0

T.F 9

pe-O II PROJECT MANAGDIENT se 4

I 1

k II. PROJECT MANAGDE!Tr In order to successfully implement a large scale research effort such as the one proposed for Crystal Ris 3r, it is essential to firmly establish a systematic plan for efficient project management. Several different levels of management and lines of interface are necessary. The model of the management plan proposed for Crystal River is depicted in Figure 11-1.

Each circle represents major entities between which direction and information must flow. Of course, each entity consists of separate levels of management which are not shown.

As indicated in the figure, the primary interfaces are between the AEC and Florida Power Corporation (FPC), and EPA and FPC. FPC has the responsibility of relaying progra= ctrategy and modifications from the AEC and EPA to the individual projects and subset tently to return the data, interpretations,

- and conclusions to the AEC and EPA. On the other hand, both the AEC and EPA have the responsibility of distributing the information received to the other agencies and of coordinating questions and suggested modifications from these agencies. In order to enhance communications, quarterly status reports will be submitted to the AEC and EPA, and as deemed necessary,

- review meetings will be scheduled.

FPC will interface directly and continuously with the principal investigators on each project. This interface will be accomplished by phone communications

. 4

l.

• and supplemented by on-site and periodi interoffice visits. In this manner the optimum flow of information can be realized with the assurance that the research is always oriented toward the desired objectives. In addition to the interface between FPC and the principal investigators of the individual projects, communication will be maintained with two special groups: (1) Statistical Control and (2) Modelling. These groups will provide valuable information relative to the specific strategy and techniques of the individual researchers.

Finally, lines of, communication will be established between each individual principal investigator and the mq/delling group. This will insure a flow of data and summaries to the modelling group in a timely and orderly manner.

This flow of information is essentd al for the utilization of this group in reaching meaningful conclusions concerning individual segments of the research

• effort and in providing a perspective by which one portion of che work can be compared with all others.

9 9

ks N

5

4 PROGRAM OBJECTIVES '

FPC l ( A EC - EPA '

DATA + INTERPRETATION ybb V

/c(

STATISTICAL $ MODELLING E TIONS S

COVTROL GRWP p*?/ o 1

// , 2- , h. c BENTHIC PHYSICAL 1 ,,,,,,,, l COMMUNITY PARAMETERS

=

T T WATER i f/SH I l PL ANKTON i 1 .

O FIGURE II-I PROGRAM MANAGEMENT MODEL t -

1

III STATISTICAL CONTROL l

l

111. STATISTICAL CONTROL The federal agencies have agly suggested the use of a bionetrician to provide advice concerning the statistical validity of the various research efforts to be undertaken in this program. FPC is, in addition, considering the advisability of employing a third party to act in an advisory capacity concerning general program management as well as questions of a statistical nature. FPC is evaluating several approaches to this problem but has not reached a firm decision at this time. As soon as a control plan is finalized it will be forwarded to the concerned agencies for their review and approval. At the latest, this portion of the program will be presented at the FPC - federal agency review meeting tentatively scheduled for about the end of June.

S N

+

7

?-

e i

}

i I

8 a

IV MODELLING FOR UNDERSTANDING t

1 I

f r

I 1

4 4- 1 i

l e

IV. MODELLING FOR UNDERSTANDING s

The modelling activities proposed in conjunction with this project serve two broad functions: (1) to provide a vehicle for understanding the relationships between major entities within each subsystem or system; and (2) to provide a predictive capability for stressed systems. The latter role, which is extremely important and is the ultimate goal of ecosystem modelling, has not at present been perfected to the degree that it will likely provide completely conclusive results in the time frame of this proj ect . The models will, however, be capable of indicating general trends and thus greatly aid our understanding of the dynamics of the ecosystems in question at Crystal River. For the short term purposes -

of this project the overview function is of greater significance.

The primary objective, then, of the modelling activities for this project is to provide a vehicle for understanding the interrelationships of the major components of the subsystem or system being studied. Each subsystem, for example the marshgrasses, is modelled or described in as complete

, a way as possible. All the major components and their interrelations are included along with the lines of energy flow which connect the components.

As data becomes available and as our understanding of the subsystem becomes more complete, modifications may be made to the model to provide a more complete picture of the existing subsystem. The modification may consist of the addition of a component found to be relevant or the deletion of

- 8 r-

a component shown to be insignificant. Therefore, the model at any time shows what we believe the subsystem to be and provideo a convenient tool for evaluating information gathered concerning specific components or interrelationships.

' After each significant subsystem is described, the overall system may be modelled by interconnecting each subsystem in a manner indicative of our I

best. understanding of the system's interrelationship. Again, modifications

.- may be made to the model as our understanding increases. Thus the model t

provides a picture of our conception of the major components and their interrelationship for the entire system being considered.

Modelling as discussed above provides a diagrammatic tool to help in understanding the system under study. In addition, our understanding may be further developed by expressing the model mathematically as a system of coupled differential equations. These diffential equations, in turn, may be simulated on analog computers. In this manner the behavior of the system with time may be calculated. There are two reasons to perform 4

such calculations. First, by simulating events which are commonly understood, the model may be tested to insure that it duplicates the natural system.

If this is found to be true then the validity and essential completeness 4

of the model is confirmed. Second, simulatiens may be performed to calculate effects due to perturbations which are not commonly understood. This is the predictive function mentioned previously. These calculations provide

~

Lindications of general trends and thereby improve our understanding of system dynamics.

9

f The models currently conceived for each subsystem and the overall system are included for the review of all concerned regulatory agencies. In order to insure that a common understanding exists between all parties involved, comments are solicited concerning the validity and completeness of the models involved. If the agencies believe that the environmental systems at Crystal River are not adequately depicted by the models then suggested corrections should be forwarded to us. This will allow sufficient time to resolve any discrepancies prior to program termination. In this manner all can be assured that a common understanding of the relative importance of each component within the system exists.

Thus, when the time arrives to analyze the very extensive data gained from the research described in this document, the analyses may be made and conclusions may be drawn in a manner which assures that the results from each specific research activity are placed in proper perspective relative to that from all other activities. In other words, this will insure that data concerning specific species or areas are considered in their relative importance in the ecosystem.

e

{

10

0 m

! M r o ,2 2

i B g $ a i L y} 3 x e5 1 ap a

- x '

2 F )

gi Tm A

g 7

/yp r

A pe .t l3 a x

yy .

' I 7 ,

'l:

i N 1 x ?u g, ,

1 h %

s[

y g

~

u-Li g

s. y

f m/ p a3 5 b$ s $,

'____..__gr'

^

i . ,  ;

O a

F e

fust.s l (uanw)

PAMS L

ANIMALS OR6Adtc.

MATren .

(R.AMK704) xoventars zco. ,,,a

= w?nx As&

~ m SustLnsa STACK 310M.4 0 '.'.

( RnEF')

$Un r s~, ss ANIMAL $ .

(BOW 1) > f/3HI^!$

Fig.IE-2 SoEe principal isotope storar.c sites in the system of Tig. g :

l l

~

IMTAKE CAMA' '

PcWER DiscuArtcys

/ ~ 's y f-qAMAL

/ WATE;t M L~

\ g } 40Cl cowciNa: nom am

/ ygoT LEVG g --

l j ,

MdT an pl r aar y i anr ,

i x a>& '

}/

I X

x I

yro- Tco - 3 1 i V / I 7 rco- PHyTo- }

I FLAuried I R.a>JrJ:>J YrDN I !h ) [ l '4K c 4 I

- 1 I Ykl , f I

l I ag3 , 7 PAod-icanaes e,acos _(

l

_ - r.meos mmrus;l

{

t r- L---[a, 'Dd"

j 5 INTAME g i/

~~

=

\ O.z l

/

1 Og g bCREEMS

\

/

L I RT  % ^) \% /

\ I y

,v, CHLORIMATio4 ._ s /

x X,, scaeza

\ b6N 2b s /  :-

%s 4

1 Fig. IZ- 3 The Power Plant Subsystem.

l Qg, d

b b b g e a-y

~ _ ~<~xx x x x x s

s 3.

P3 i

v N -/

9 ~; '

1

.b

\q\

m2 l2

• l

~

$E b

, di f ^ 6\\Y. f\h l W

)

7 /d g9 t

• 's O g

' ' y n 1g

+ "

t  !! \

hs Y N" 1

\ N f o li M.

ya

$,8 i \n!" -

c 8, .If

..g<> -

5 O w ~#

Q l34 U 0

h Yl ( g

~

z / C

(~~ke +

< 3 1 e iTV N s/p/s/N/, &

= 1- %

c t ^

G 1 d 'x$ fe i . i~

A I

~['

s 3 l [a p ~

/ By .

N C

- / Ec f .. - ,

/

\

5 x

66 a QT E If z

o I =

N 6 F / 4 4

\ $l x

G

/

Q

%~ ~~~____ E_ #

f n

1 l

,s, j

/ ,v, =

~ , a>> . - l1 s sie l /

.+L ja f

$I s, .

^

/

J ffj

/'Q ff-s '

!I j

j; 4 3 -

ga\

1 l

awd f'g: ll

[

s /h a p I Isl 3 - No s -

x gi i w o- - + \ $ f

• i

? -^( *l G -

n N

l b 's _

'f ,,'n, e

} .

4 w

- ) E 9 'a x 15

h O

$5 . EE qi " 2 05 g ()s  :: x .u :e z  :

.x

?

> , :x erses

• 3%

lit

~ ~

• fs '( 5

?.s EEE $ 8 ?55 *u

" E & " % # # R Te i 2184554 u" ~

Yb' N'  ? J2Jau .

h. E ok pu e'A V

x 44i 6

\* ;3 E

s,un L e- } .

e, 4

= - ,

a ,

k

-)

^

k / p eP ,

4h i

5 e,

e e

'h g's, w-

+- h a E < f I

N s N / A ,4 m 5

e q

4$s

D

- x:

c[7 [ $7 i n . 1 4 85 M r.. ~~

x- i j

j ,

/g =

,* SM, i by\,g &'. ,y 2 ~

, }

m n

-c o y4 A b' 5 i'E $$] '

6 41 E' d'6$ 3 u

^ ^^ n o$w 1 -

n ea e "* E I e

• C c' t E l 5 E

• I D

lI -S C E B

• ' R$

2x x x < 7; a -

/

/. . 4,-y=

~

(

. -c%., s

/ >

a d E' S Y

f

- n a \1 ^ s l

[k,I p l~ ~P l ,

^

f.

.W .

. 6 j

s

~ - ~ ~

~ ~ ~ ~ ~ ~ ~) ,

a b1I M o

=

. / 2 5y E3

~~

"/

x x x7x >d <,,,

G

~  ;

C C h i  := a l' 6 i x8 f-l 1 O@i 17

~%

1 l N l

7. a ,

i l i fg ] .m7m oa MAR 3M

05. Og. X l x l <

f ) 3 7 l x

y N.-

% y _.

\ / G $ NAILS

'/HHER BA'r' CRASS,f

\ insec,s*c ST4 Auss aizac, 1

\ \ MEMT

% Sug

/ bio y rsss M

p B.D f 1- '

D hm0 turCRIAL l .7 QH R

NARS// h 4 4 y,gp N/4HER PM HecaoerL Y4/4: ( Symbol Key -

% M -'[ Org - Organic Matter f .

P - Phosphorus T - Temperature

% Biomass:

4 Y - Young plants gg) M - Mature plants Rm D - dying plants i Y Rrorst Fig. II-B Energy Diagram for Salt Marsh System.

't

~ __ ._ _. _ . _ . _ _

2 i

1 h.

1 5

i s

s V

PROGRAM DESCRIPTION DW d

b f

7

(

V. PROGRAM DESCRIPTIOM The environmental research program described in this chapter conforms as much as possible to the AEC and EPA proposed programs. In a few instances relatively minor details remain to be resolve . These are being addressed currently. In some cases the program requirements w' found to be unrealistic. Chapter VI of this document has been included discuss such inst,,ances. In cascs where exceptions to the program were taken, as many experts in the relevant area were consulted as possible within the time frame in which the program was formuisted. As a result, we believe that this program presents a very comprehensive, well organized effort which will facilitate an evaluation of any reasonable cooling alternative for the units at Crystal River.

For ease of presentation and understanding the estuarine area adjacent to the plant site is. described in four regions: (1) intake area, (2) intake canal, (3) discharge area and (4) marsh grasses. The research being conducted in each area is discussed according to subject.

A. Intake Area

1. Physical measurements to determine the source of cooling water.

a) The objective is to determine the source of cooling water to establish the origin of the plankton population being entrained.

N m 19

b) -Techniques employed will include:

1) Long term (up to 6 months) recording current meters positioned in the intake area; 1

2) Periodic (bi-weekly) samplings with a portable current meter covering sufficient stations to give good source resolution;

3) Dye studies to determine flow patterns and mixing of intake waters; o

4) STD surveys to determine water characteristics in order to facilitate identification of water types;

5) Radio-beacon ~ drogues to trace water path-lines over a tidal cycle.

c) Runs will be monthly except as indicated.

4 d) Station locations are shown in Figure V-1.

2. Plankton Sampling a) The objective is to establish the species being entrained in the system and their quantities, condition and fate.

, 20-

r.

E' b) Techniques (1) Sampling Program (a) Zooplankton (i) The techniques discussed in " Environmental Research Program at Crystal River - A Technical Discussion" P. 46 will be used.*

(ii) Sixty-five (65) gp net hauls will be taken periodically to determine correction factors for the biomass missed by larger nets.

(iii) Replicate samples will be codbined for counting.

A portion of each replicate will be saved for contingency analysis.

(iv) Two replicate samples will be taken at each station.

(v) Surface current, speed, and direction will be determined.

• The report cited was submitted by FPC to the AEC on 3-6-73 and discussed in detail the research program currently underway at Crystal River.

W 21

(b) Phytoplankton (i) Routine sample size will be one liter. Samples will be preserved in 5% formalin.

(ii) Settling tubes and an inverted miseroscope will

~

be used in sample analysis.

(iii) An immediate examination of live material will be

_ made monthly at selected stations.

(iv) Phytoplankton primary productivity will be measured monthly by the C14 method.

(2) The fate and condition of organisms will be determined as outlined in the proposal submitted by Dr. Fox and Dr. Maturo (See enclosure C,Section VII).

c) Station Locations

- (1) The locations of sampling stations are shown in Figure V-2.

(2) The rationale for station locations is:

i 22

). = w t- (a) Station A gives a measure of inshore benthic invertebrate larvae and ichthyoplankton populations.

(b) Station B gives a measure of plankton in area 2 which is currently concluded to be the principal source of cooling water.

(c) Station C gives a ceasure of plankton in water similar to off-shore Gulf water.

d) Frequency of Sampling (1) Station B (a) Quarterly, the station will be sampled hourly over a 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period. Only ,_lomass determinativn and the identification of major species will be performed.

, (b) Every four weeks a 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> sampling will be conducted based on tidal and diurnal cycles as determined by methods discussed in paragraph V. A.2.d.1.a. of this section.

(c) At four week intervals, two vecks out of phase with l

the procedures in the previous paragraph, a surface I sample only will be taken.

23 h

l (2) At Stations A and C a surface sample only will be taken every two weeks.

l l

i

3. ' Water Quality a) The objective of this analysis is to aid in determining the origin of the cooling water and to determine the nutrient levels of the cooling water for background information.

b) Highpoints of the techniques are listed below.

(1) Samples will be taken concurrently with the zooplankton sempics.

(2) Salinity and temperature will be taken in situ.

(3) Nitrate, Nitrite,~ Phosphate, Ammonia and Silicate will be analyzed by a Technicon Auto Analyzer.

~

(4) Dissolved carbon will be analyzed by a Beckman Analyzer.

(5)' Chlorophyll content will be determined by an acetone extraction.

4 24 l-

l i

I l

(6) Suspended load is to be determined by filtration of a water sample and gravimetric methods. .

c) Station locations are the same as zooplankton stations.

d) Samples will be taken during zooplankton sampling and primary productivity measurements.

B. Intake Canal

1. Physical Measuresments a) The objectives are to determine velocity profiles (over various tidal conditions) at cross-sections located along the intake canal and at the intake screens, b) Station locations will be evenly spaced over the distance from the intake screens to the end of the confined channel.

There will be three stations over the span.

c) Frequency of sampling will be over two complete tidal cycles twice a year.

-2.- Plankton Samplings a) The objectives are:

25

(1) Similar to those outlined in paragraph V. A.2.a.

(2) To determine similarities and differences, if present, with the intake area water.

(3) To determine the contribution of the canal population to the estuary.

(4) To supply input data for the ecological modelling effort.

b) The techniques to be used are similar to those discussed in' paragraph V.A.2.b. In addition at Station E, three levels will be sampled. Use of a pump is being investigated.

c) Station locations are shown in Figure V-2. The ratienale for station location is:

(1) Station D measures the population entering the canal system.

(2) Station E measures the population being entrained.

(3) Station F measures the population leaving the canal system.

(4) Other points as outlined in the Fox-Maturo proposal (see enclosure C of Section VII)..

'26

d) The. frequency of sampling for all stations is the same as that

. outlined in paragraph V. A.2.d.1. for Station B.

3. Population of Canal i

a). The objectives of this study are to determine the population

- characteristics of the intake canal and the beh'avior relative to time, tide, season, barge movements and intake velocities.

b) Stations will be located along the length of the intake canal depending upon the , type of net employed and, in turn, the portion of the population being sampled.

i 1

(1) Eight Uyoming fyke-nets are to be employed along a 0.5 to 1.0 mile portion of the canal to determine net directional movement of bottom organisms.

(2) Sixteen Coffin traps are to be employed at 500 ft.

to 1,000 ft. intervals to obtain relative population densities along the canal bottom.

(3) Twelve gill nets of varying mesh are to be employed along the canal banks banks and extended into the canal. These nets will be used to determine trends in movement of mid-and top-water species.

m 27

(4) Tagging and marking studies will be performed in conjunction with the sampling described above to help determine movement patterns and give an index of species density in the intake canal.

c) The fyke-nete, Coffin traps and gill nets will be deployed sequentially at one week intervals during the sampling period.

4. Fish 1mpingement Study a) The objective of this study is to quantify in terms of number, size / age class and biomass, the species impinged on the traveling screens.

I b) Samples are collected at the ends of the screen wash. The samples are then sorted to species, counted, weighed and lengthed. Statistical evaluation of the data will be performed. Both outfalls will be monitored simultaneously to determine the variations between the outfalls.

c) 11ourly monitoring of impinged organisms will be performed I

on a weekly basis. This program will be continued until 1 full year of data _is obtained. The screen-wash monitoring l

l I

i

( 28

, e

.will be continued for four weeks beyond the one-year record to obtain a data overlap. In addition, sampling of each end of the screens will be performed for three consecutive weeks.

S. Water Quality a) The objectives are:

(1) The same as given in paragraph V.A.3.a. of this section.

(2) To measure changes in nutrient levels caused by entrainment.

b) ThetechniquesusedaregiseninparagraphV.A.3.$.

c) The station locations are the same as those discussed in paragraph V.B.2.c.

d) The sampling frequency is identical to tre zooplankton sampling discussed in paragraph V.B.2.d.

6. Diversion Studies The' University of South Florida Department of Marine Sciences, is currently investigating the use of artificial and 29 l

na: ural barriers to the free movement of fishes, spillways,

~

fish ladders, and other methods of diverting fishes in an attempt to solve the problem of fish entrapment and impingement at the various Florida Power plants. This work will be coordinated with the entrapment and impingement studies proposed for Crystal River in paragraphs V.B.3. and V.B.4. of this section.

C. Discharge Area

1. Physical Measurements a) The objective is to define the existing three-dimensional thermal plume under various hydrological, meteorological,

, and tidal conditions.

b) Station locations are shown in Figure V-1.

c) Techniques will include:

(1) Long term recording current meters; (2) ' Dye studies; ,

l (3) STD surveys ~ to determ'.ne water characteristics in l

l ,

order to identify plume water, Withlacoochee e

i 30 t

c

fresh water, and Gulf. water and to define the thermal pit s.c its zones of mixing.

(4) The federal agencies have suggested the use of thermal imagery for plume mapping.

Thermal imagery overflights of the Crystal River Plant thermal plume were performed by Cornell Aeronautical Laboratory in 1970.

The results of these flights were of little value since it was demonstrated by field measurements, that the plume sinks rapidly upon leaving the plant, and is not discerned by infrared photography.

d) The frequency of surveys will be quarterly.

2. Mathematical Modelling a) ,The objective is to develop, verify and/or modify the

~

the'rmal plume mathematical mor e A to accurately simulate i

the plume described by the work in paragraph V.C.1.

i b) Refinements include:

1 31

_(1) A program called " Beer-Can" has been added to simulate the movement of a water parcel, drogue, or similar tracking device.

(2) Watar storage of marsh-creeks is being incorporated into the model boundaries.

. 3. Plankton ~ Sampling a) The objectives are:

(1) To compare stations in the thermal outfall with stations in the intake and canal areas.

(2) To establish a station in the thermally affected area.

(3) To provide data which can be integrated into the modelling of the inner and outer bay ecological systems.

b) Techniques are identical to those described in paragraph V. A.2.b.

c) Station locations are shown in Figure V-2. The rationale

.for the stations is:

(1) Station G measures the area of the estuary receiving the thermal impact from the plant.

32

(2) Station H is relatively unaffected by the thermal

. plume and will show if the population in this area is similar to Station C. It should be useful for evaluating primary productivity.

d) Sampling Frequency (1) Station G will be similar to the intensive sampling program outlined in paragraph V.A.2.d.1.

(2) Station H will be sampled at the same frequency as Station C, see paragraph V.A.2.d.2.

l

4. - Water Quality a) The objectives are to determine water quality for:

(1) Incorporation in estuarine modelling.

(2) Comparison with the intake area.

b) The techniques to be used are those outlined :bi paragraph V.A.3.b.

c) Sampling stations will be the same as those identified in paragraph V.C.3.c.

33 g q, .- y- , ~- cy ,-- - , - --- -- ,

4 d) Sampling frequency will be the same as discussed for paragraph V.C.3.d.

5. Marine Environment Survey a) The objectives of the survey are:

(1) The assessment of thermally-affected and non-affected portions of the study area.

(2) The measurements of abundance and distribution of benthic macrophytes and periphyton, microinvertebrates.

(3) The quantification of per-unit-area and total area ecosystem components.

(4) The preparation of base maps of the ecosystem by Se8 Son.

(5). The preparation of map overlays of physical and biological parameters.

b) A description of the sampling techniques to be employed is' given in Table V-1.

c) Station locations are controlled by the diversity of bottom 34

types. Three areas of different sampling frequency are described below:

(1) A trapezoidal-shaped area adjacent to and north of the discharge canal has been sampled quite intensively because the diversity of bottom types precludes stratified sampling based on substrate and plant community. Most of the work has been concentrated in t. fs area in the past since the potential effect of the plume is greatest in this area.

(2) A region enclosed by the 8'F isotherm as calculated by Battelle and shown in Figure 5.3 of the AEC's Final Environmental Impact Statement dated May 1973.

(3) A region enclosed by the 2*F isotherm and shown in the reference above.

In the latter two regions, investigative sampling is being conducted to determine if the region can be readily categorized by substrate and plant community. Since the depth of the water in these regions precludes the possibility of identifying bottom types from aerial photographs, - fifty stations will be located and sampled initially. Hopefully, the information 35

E.

l l

gained from this sampling will give light for ways to improve the method.

6. Sedimentation Program a) The objectives of the sedimentation program are to:

(1) Determinetheeffectsofnon-catastrophicde/positionof materials on benthic primary productivity.

(2) Analyze the existing sediments to determine particle size-and composition, b) The techniques to be used are:

(1) Varying amounts of sediment will be placed on plots and the plots' productivity will be measured by noting differences in oxygen production occurring in an enclosing dome.

_ (2)- The degree of sediment analysis has not been determined at this time. It is expected that particle size, settling rate, carbon and nitrogen content will be measured.

Details will be made available as negotiations proceed.

36

D. Marsh Grass Study .

1. The objective of this study is to determine the effects of artifically heated water on the marsh grass community.

2. Techniques a) Primary Productivity and Marsh Grass Nbtabolism

. 1. Annual net community production is measured by the harvest method for live and dead material within 2

a 0.25M clip quadrat. Nine samples are taken of the Spartina and 5 of the Juncus. Control and 4,

af fected' areas are compared.

!< . Grass community metabolism - Total community metabolism is measured by measuring CO change in an environmental chamber. Four replicates are sampled continously over a 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period for a five to seven day period.

b) Population Dynamics The major consumers in the marsh community, snails and fiddler crabs, are counted in each vegetation quadrat.

~

3.. Sampling Areas 37

The affected area being measured is the marsh area immediately north of the discharge canal. Two areas are being used for controls, Lattrel Island which is a mile north of the canal and Negro Island which is to the south.

4. Sampling Frequency a) Community metabolism will be sampled quarterly.

b) All other parameters will be sampled monthly.

4 E. Other Areas Incorporated within the program proposed by the AEC and EPA were requirements for general surveys in areas potentially affected by alternative cooling systems. Most of the areas of concern have been addressed in the program discussed above.

However, plans for some of the surveys, in particular, those dealing with the tc restrial flora and fauna and the nursery areas _ have not been completed at this time. These plans are

. being addressed currently and the finalized plans will be made available to the AEC and EPA at the first review meeting.

~38

-- ~ - _  ;

TABLE V-1 FIELD SAMPLING TECIINIQUES To be used in the themally-affected discharge area and control area to the south of the intake canal Semi-

• quantitative techniques will be used to survey the much 1arger-area under the potential impact of the thermal plume.

QUADRAT SEDIM'iT CORE VEliT'URI PUMP DROP tlET TARGET ORCANIS}tS macrophytes and micro-in' titebrates macro-invertebrates vertebrates and

, periphyton macro-invertebrates PRD%RY OBJECTIVES biomass and relative abundance biomass and biomass and diversity and diversity diversity diversity SECONDARY OBJECTIVES time and s,, ace time and space time and space time and space distributions, distributions, distributions, distributions, phenologies .dietaries and dietaries and dictaries and

. life histories life histories life histories SAMP!.E AREA 1 m2 ~78.5 cm2 , 1m2 .

16 m 2 .

SAMPLES PEP. SEASON 200 100 26' 54 IRN-DAYS PER SEASON 42 6 30 81 CEAR m2 frame and coring cylinder, n 2 enclosure 4m x 4m PVC frame, bottom scoop 10 cm dia. x frame and 18mx2.Sm, 1/16" 14 cm ' , centrifugal- mesh drop net and

. pt:mp driven n,eeping seine venturi suction Pump

. e G

O 9 I

I l

39 j u .a

,p _ _ _ ..

r --

p * .',,, j '

% \. .. . . . ..' .& <

rp,h,s ' -

'E8,'}- sJf* * ' **~..."M. - .

./ g p,',. .;;;. . .

".' Q ~ lp ,g c5(f6 '

lN

\ . c, ..-- -

\ s. . . . . . sw.v. . , , ..

h >  % -

<. . . . , . . .t--..

• e

.s =. m

-. S.

. 8 ,

. .:.7. .. .

'g. .. . . . o .

W. .f . _

) .0. g .

. c= . -e o

.s-sf. . :-

.: .  ;. * ,s. ;;, g.. J ,

,., ."* A A0 #*-

(s

.c= , o es0 6 .

\ .' ' ' ~..

~'

z,

. . . . w.l. Afg :s Am y

W d *

.ew)(u'-:l

. .' y &

y. . . .. b

.N .A

• o $ ,

.. 4.. .<. e b o% / W

-). ..,' ' .' ,. =@g..,

,9-

• p.

• 3\

• o

% I

},

. d' k i 4 A

\ *,p ) r.q at Q

' o 1

j 4 g

y A %.

. . 3y

. s  !

\, .t ~

..g"R.

.s

"" 4,..s'h. ~ ,

t

• ' ,/ .uk}...... \ fA....h :* S)js'J >i e A CtAWRCNT . ., *Aj.\

ranx.,STAft0NS ~.

=

. ~ a r,T,,-

< 0%

e.ees

)

E rDf R[COMOING STATIONS e PfRMANCNT sig g rattox$ ,

,4 ,,-

• %s ) .j >.gh

),*

0

,.. -., . ort sticasc SrarrQus  % p f

s.s,r> an e s. m

, , _.y ,

g y q,,, -

s, e .r. s. ;y .r+

I 4

,. / s J +

$ ' *p,

. ( ;

. . . . . . ~ . .

}

. . z.

5 'S{

.\  ! \$ #-

1. . . . . - ,

) [' ,'> \\%. ' . ' B Figure V-1. Nermal Plume and intake Source Sampling Locations.

40 .

e-

OFPC EXPANDED SAMPL/NG AREAS .

O O*'  %+: , pg y -;- <

cv c R & TATION F

. j

+.

3 d- 1O .  ;

v STAT /ON G L u" %J STAT /0N H

'::. ' -='? " e.Y SN'% pY oo I s

.  :~

A l' .) STATION E s

I STATION  :.

D ';

s i .

g-s .- .. :: g., y

-C ' S'4 ...~

• ?:. ,.F.i

'.;.STA T10N A'

+. V q

l s

Cj' '

'~~i,<. . .

,.s:

o.. q.. ,. , . . .

,r

?:i

• . . ' STATIOU B S.. . ii

\.::*

1. , a -

. g i

c r.: ,.$

I

.s . -

e-15 Q I  %,*;. .

",f CRYSTAL

1. O RIVER STAT /0N C ! gi. }i  !.' i.

i .:

l s ..

. 9 AREA 3 l, -

AREA 2 /'- . ARdkI:M/::-).

s 1:

I::: ......

,  ?. +.% .

(

A i

n l,.2:

.-.# Ti

-- c; ~. v.

E s I. c. .

l V. :  ::<. :is I \ ." - eq I .*.j -

\*] =  %

l

, fr. r.

Q:.

C- -

-e

, . s:: -

.O i* 2 fi: .f.: :~ .

~

1~

:  :? r. :J NAUTICAL MILES 10 p,i!

. y . ': ' -

Figure V-2. Plankton Sampling Stations.

V VI Exceptions to the Program Proposed by the Federal Agencies t

u_

9 A. Plankton Program That portion of the specifications and guidance from the AEC, EPC and other federal agencies pertaining to plankton was forwarded to experts in the field of plankton sampling and analysis. They were asked to comment on the feasibility and practicality of the ,

federal program requirements. In addition, Dr. Maturo of the University of Florida, the principal investigator of the presently implemented zooplankton study, was asked to develop an implementation plan for the federal requirements.

Some of the questions which arose during the review of the federal requirements are discussed below.

1. The Federal program was unanimously concluded to be much more extensive than necessary.

2. No method for determining fate and condition of Planktonic organisms was given. No established procedure for this determination is known to us. The method outlined in Enclosure C of Section VII is being submitted for review and suggestions as an alternative.

3. No means was given as to the way the mass of data obtained from such a program could be interpreted. It was suggested 42 m

by some of our reviewers that a modelling-approach would be the only feasible way to approach interpretation of the significance of the data obtained.

4. The (MARMAP) methods outlined in Ahlstron, Elbert H. ,

Kenneth Sherman and Paul E. Smith (unpublished MS) titled

" Sea Going Operations" appear to be applicable to water deeper than 20 meters. While we feel that similar equipment would .be useful for comparison with offshore areas, the shallowness of the area under study will preclude their use as a valid sampling tool.

5.- Finally, and most importanky, the manpower requirements for the federal program are far in excess of what can be assembled in the time frame of this program. The number and approximate locations of the sampling stations are

, shown in Figure VI-1. The actual number of samples to be taken at each station is summarized in Table VI-1. While

~

personnel can be obtained to collect samples at the given frequency, the area is almost void of tehnicians who are suitably trained and-motivated to analyze the samples.

Dr. Maturo has . estimated an 82-person work force to analyze the samples. This estimate is based on his current work which requires one man-day per sample and yields biomass information and some idea of species composition. Our 43

best estimate of the number of qualified personnel available for this program is approximately twelve. Due to this personnel factor the sampling frequency will have to be restricted. The program outlined by FPC in Section VI of this document can be implemented with the personnel available. We recommend its acceptance as the best possible means of attacking this Plankton program.

Because of the reasons outlined above, the program outlined by FPC has the following recognized restrictions. The program will not necessarily document rare events, those occuring less than 1% of the time. Neither it, nor the federal guidelines, will give a boundary to the planktonic population being entrained. Finally, the " Fate and Condition" experiments may not answer those questions.

We will welcome third party arbitration of any portion of the plankton program. If the interested agency personnel have objections to the program they are invited to submit alternatives which can be evaluated by disinterested, qualified professionals in the area in contention.

If scientific validity is not the criterion upon which to base evaluation of a program,. as was pointed out by Dr. Prager of the Environmental l Protection Agency, it is requested that the specific legal references and court decisions upon which the program should be based be forwarded l

44 A

O PROPOSED EPA /AEC SAMPL/NG AREAS O

%'H"O c

O** "% , re  %

b> M.[~[f,f STAT /0N G l V Q STATION F Y C

[]h

$ i

. ,cb Q /" _ '*** _

o* I

?.: ' STATIONg Hr'iLnt 4 W' QJ,,

o a ' '.:-

g .

, SMTION E

O !, STnp ON D t-

\

I ..

. r O. v' ', -

9. .:.:. .

~

C l C STATIOf "')_',i.

, '?  ;.:i_:.STATKON A y STAT /ON J 0 l C 5%:.:.

~: d'h.9.:.:,:.(u),s,...

,('

s/  :: .. . .

.. u

, STATION B ' s,. .  :;  ; \::; .. !

"". , r. .

q

1. O'

~-

I I

V:.

v.

?!

.}  :

I

.:; os.j.A so.. i CRYSTAL niye :

D, B '

STAT l0N I,C \, i? . .

w ..: s:- 9 AR5h I ::'y#.e I; AREA 3; > c,i ,i

. AREA 2 i[. -

- I;.e;

.: ~ :. ....

v.gv 6

$ l 0 h/gI1 l:'  :?

? '1 i' s.:

W.

'T , i.'s I. (

l"

(. :. .i.: .

)$',: tl IhI *: .

(- 2)C"! l r. - C.-..

ii 1-::

4

. I:

a i

9 1 -

2 ti::.; . *s . .r.: -: .

i:  :-r . e.:

NAUTICAL MILES I:t p,r:

. y -

N rrcuas VI-l EPA /AEC SAMPLING STATIONS

TABLE VI-1 ZOOPLANKTON SAMPLES TO BE PROCESSED PER MONTH (ACENCY PROGRAM)*

Station Identification Diurnals Sampling Events Number of Number of Samples and Number of Depths Replicates

Stations M nth Sampling Period Subtotals Total A,B,C,H,I and J (6)

. D&C 4 8 3 2 384 (2)

E&F 4 8 1 2 128 (2) 704

• Based on Agency Document titled "AEC Related Environmental Research Programs at the Crystal River Power Plant Site" pages 2 through 6, Articles I. A through D and page 8, Article 2.a and the assumption that only two re-plicate samples are required.

NOTE - THIS DOES NOT INCLUDE SEPARATELY COLLECIED ICTHYOPLANKTON SAMPLES.

i to us imediately. This will reduce the possibility that the study will not produce the required information and irreplacible time and research staff resources will be lost during the most critical data taking time fra:se.

4 47

/

B. Fish Impingement Program .

Florida Power Corporation initiated a comprehensive screen-wash program in t-u gus t , 1972. The objectives of that program were to quantify in ten =s of number, size / age class, weight, and condition the species which become impinged on the traveling screens.

Scientists which are responsible for this program consider their current sampling frequency schedule to be adequate.

Differences in Current Sampling Programs 5 Current Fedaral Guidelines-(1). July, 1973 to July, 1974 4

(2) Sampling at both ends of screens.

(3) Frequency - every 3 hours3.472222e-5 days <br />8.333333e-4 hours <br />4.960317e-6 weeks <br />1.1415e-6 months <br /> for a 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period, twice a week.

Florida Power Corporation l

l (1) August, 1972 to September, 1973 , j O

b 48

- (2) Sampling performed on both ends for three consecutive weeks to note variation.

(3) Frequency - Every hour for 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period once-a-week. Data overlap for one month will give additional information.

We believe that the program already underway with the noted modifications will answer the questions raised by the federal agencies. As the data becomes available for review, further modifications to the program can, of course, be made.

n

.49

s.

4 3.

k i

VII ENCLOSURES A. Interagency proposed program B. University of Florida evaluation of the interagency's plankton program.

C. Proposed program to determine 1

1 fate and condition of plankton.

(Drs. Maturo and Fox, Univerity of Florida)

J i

I

--. ,x-- .,, _ . - _ - - , , ._

- , , , _ , . .. . _ _ , , . - . _ _ , - - , _ _ _ . . . - . . ~ .

(

7

f. .

9 g..

I e

ENCLOSURE A 4.

)

s.

i-

)

AEC RELATED ENVIRONMENTAL RESEARCH PROGR/J.S AT THE CP,YSTAL RIVER PO'a*ER PLANT SITE .

Problem

• To determine the need for modification of the proposed cooling system for Crystal River Unit 3.

Purpose

' 1. To obtain necessary data of the Crystal River Site area from a' coordinated and comprehensive hydrological investigation.

2. To identify and quantify those factors that have impacted the Crystal River environment and to obtain necessary information on aquatic organisms and water chemistry in the Crystal River Site area in order to be able to assess the potential impact *'n o the aquatic bio'ta from the opera-tion of Unit 3. .

Objective .

To provide a basis for a decision with regard to the need for an alter-native cooling system for Unit 3 no later than November 1974.

' General Discussion ,

The AEC staff, in conjunction with other interested federal agencies, requires additional information in order to predict the incremontal impact on the aquatic biota from the operation of Crystal River Unit 3. Of necessity this assessment must be based on data collected in conjunction with the operation of the oil-fired Units 1 and 2. The specific areas of

, concern are hydrology in the immediate plant environs; entrainment of organisms through the condensers; impingement of organisms on the intake structure; entrapment of aquatic organisms in the intake system; thermal,

~

chemical and physical impact in the discharge arca; and biota surveys in g 4

. 50 .

~

a

't areas which may be affected by candidate alternatives to the proposed once-through cooling system.

In conjunction with the study program required in each of these areas, the applicant will concurrently initiate and complete detailed hydrological-environmental assessments of alternative cooling systems to identify those systems which would impose the minimum environmental impact, taking into account the areas of concern expressed above, including terrestrial impacts which are not involved in'the proposed once-through cooling system.

The applicant should levelop a study program in accordance with the recommendations and guidance developed in this' document and will submit this program to the AEC for evaluation. Such evaluation will include a review and consultations with the interested federal agencies.

Within 90 days, the applicant will submit a progress report on thi s study. Following this submittal, a meeting will be held with the interested federal agencies to assess progress, results, and evaluate the need to modify the, program. - -

Specific Procrams I. Entrainment A. Objectives .- '

l

1. To determine the source (s) ofJcooling water under normal l

hydrological and meteorological conditions and variations during high fresh water runoff periods and during unusual tide, wind and other conditions.

i l l

f- 51 .

i

2. To determine the source, fate, quantities and conditions of

- species of plankton, fish eggs, larvae and juveniles passed through the condenser cooling water system.

To determine the relation between the species composition of the cool'ing water sources as established in item 1, and the planktonic species of the intake canal.

B. Procedures for ooplanliton and ichthyoplankton

1. Length of program: A minimum of 12 consecutive months of i datawillbecollectedandanalyzedpriortol$vember1974.

2. Sampling stations: 3 stat-lons will be established in each of the three areas shown in Figure 1 (p. 54 Environmental Research Programs at the Crystal River Power Plant - A Technical Discussion); two stations shall be established in the intake canal, one directly in front of the intake pipe and another within the canal near the mouth of the double-

' diked section. .

3., Frequency of sampling: Samples shall be taken every 3 hours3.472222e-5 days <br />8.333333e-4 hours <br />4.960317e-6 weeks <br />1.1415e-6 months <br /> over a 24-hour period, weekly at the two stations in the intake canal.

Samples in intake areas'1, 2 and 3 shall be taken every 3 hours3.472222e-5 days <br />8.333333e-4 hours <br />4.960317e-6 weeks <br />1.1415e-6 months <br /> over a

~

• 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period every two weeks. All samples shall be taken to determine species, abundance, distribution and condition according to tidal stage, i

day-night variations or other pertinent environmental factors. l

4. Techniques: Replicato sampics will be taken at all stations. l 1

l Samples taken in the mouth of the intake canal shall be at surface, mid and bottom depths. Ichthyoplankton techniques will be the standard INS (MARMAP) methods and approved by the AEC staff. l e

52 7

~

C. Procedures for source of intake water  !

During the first 3 months, the source (s) of water that are  !

drawn in by operation of the plant under normal hydrological conditions will be determined. This program should consider, but is not necessarily limited to: dye and drogue studies; flow and direction studies; and calinity, temperature and water chemistry measurements. This program is to be. continued as necessary to determine source (s),under abnormal hydrological conditions.

D. Other- -

Phytoplankton studies should be carried out concurrently with the chove programs to allow quantification of species abundance, distribution, condition and total biomass of phytoplankton' species being entrained.

II. Impingement / Entrapment

'A. Objective To quantify in terms of number, size / age class, weight and con-dition the species which become impinged on the travelling screens. The study will determine the variation due to season, time of day, tide, general climatic conditions or other factors. In anticipation of higher velocitics caused by Unit 3 and the possibility of a, change in the species composition of impinged species, studies will be performed to relate the proposed flow characteristics to impingement of species.

B. Procedure to assess impingement .

1. Length of program: A minimum of 12 consecutive months of data '

will be collected and analyzed prior to November 1974 4

e 53

~

2. Sampling stations: Collections will be made at both ends of the ccreen-wash sluice until it is decermined statistically that there are no differenecs in the species composition and quantities collected at either end, af ter which collections may be made at one end.

- 3. Frequency of sampling: Samples will be taken every 3 hours3.472222e-5 days <br />8.333333e-4 hours <br />4.960317e-6 weeks <br />1.1415e-6 months <br /> over a 24-hour period, twice a week. General monitoring of the collections

. in the trash baskets will be conducted during the remainder.of the week in terms of large numbers'or biomass of individual species or large total.

i numbers or biomass of many species. Sampling and-screen operations will be modified during peak impingement periods.

~

. 4. Technique: Screen washing will be performed manually during the sample period and not on the basis of a pressure, differential (clogging).

C. Orgar. isms in intake canal Once every two weeks sampling will 'ue conducted to determine abundance, size (expressed as length / frequency), distribution and condition of fish

. species with a frequency to establish variations due to weather, tide,

. , day / night, or other factors. -

D. Diversion techniques Studies' of means for returning impinged species, to the Gulf, or

, diverting organisms before reaching the intake structure, shall be conducted in conjunction with the impingement program.

E. Other -

1. T'he number of pumps in operation and volume of water pumped shall be recorded at all times when sampling is conducted.

l

. 1 j

54 ,

2. Flow and velocity at the travelling screens, under varying -

operational conditions to be encountered during times of sampling shall be determined.

3. Vertical and lateral velocity profile data will be collected at selected sections located 1[gitudinally along the intake canal during '

an entire tidal cycle to establish flow and velocity characteristics in ,

the intake canal.

4. Condition of living impinged organisms shall be determined to establish the potential for returning organisms to the ambient waters of the Gulf.

III. Thermal / Chemical impacts ia discharge area

• A. Objective

1. To define the' existing three-dimensional thermal plume.

'2. To develop, verify and/or modify the thermal plume mathe-

~

matical model to simulate the plume described in A. 1. above.

3. .To utilize this matematical model to predict the thermal plume under all modes of operation.

4. To establish ba'seline data for estimating thermal effects.

5. .La determine how large an area of the, receiving water will be affected by modifications resulting from condenser passage.

B. Thermal plume pattern in the discharge area

1. Thermal imagery overflight information should be provided to establish the extent of the thermal plume from Units 1 and 2 and should cover varying conditions of tide and weather.

55

• 1 c ,.

~ .

4

2. Temperature and salinity measurements should be performed vertically and laterally throughout the thermal plume and should include '

1 continuous measure.ments. at the canal terminus, near shore areas and other selected points.

~

C. Water chamistry measurements should be conducted in the taixing zone to establish present characteris. tics and composition.

. D. The mathematical model of the thermal plume will be verified and/or modified in accordance with the above information and utilized to predict future plant configurations and temperature characteristics to allow biological impact assessments. .

E. Thermal / Chemical effects on biota The laboratory and field research program as outlined in Table 1, p. 89 of the applicant's progra'm will quantify the abundance and dia,tribution of macrophytes, macroinvertebrates and vertebrates. The' exact number and location of these stations must be carefully coordinated in order to obtain the maximum usable data.

1. Consolidate existing data, and supplement as necessary to develop baseline benthic survey of community structure in the projected O '

discharge area defined by the 2F iso therm in Fig.. ,5.3, D.E. S.

a. Substrate Develop maps based on particle size, organic content (ashfree dry weight) and depth of deposits. ,

i 6

h ss -

e

b. Vascular plants and macroalgae Quantitative and qualitative characterization including maps delimiting the dominant plant communities.

c. Benthic macroinvertebrates Quantitative and qualitative evaluation.

Suggest stratified sampling design based on substrate and plant communities defined above as well as temperature increments defined by thermal plume predictions.

2. Pelagic Surveys

a. A program similar to that for the intake side to charac '

s terize, predominant species of phytoplankton, zooplankton, eggs, fry, and juveniles. .

b.

, Document species composition and relative abundance of

! finfish and shellfish.

' 3. Intensive sampling in the plume area defined by the SF

, isotherm shown in Fig. 5.3, D.E.S. is required to identify species, seasonal abundance, in relation to thermal intensity, nutrients, of: .

a. Zooplankton >

b.- Phytoplankt'on .

c. Eggs - .

d. Fry and Juveniles '

e. ' Adults IV. Other general :. rveys and surveys in areas potentially affected by alternative cooling systems.

A.- Objective

' 1. To survey areas potentially subjected to impact from alterna-

- tive cooling systems. -

l .

.. 57 ' .

, .n . . .n--

2. To survey areas of interest in assessing impact.

B. Intake area -

An inventory of re:ident organisms,,especia'lly the benthos, to

~

allow for. assessment of impacts of possible changes to the intake canal.

C. Discharge Area -

Studies in III.E.1. should be extended to include expected area impacted by any anticipated extension or other modification of the dis-a charge canal.

D. Thermal effects on marshland to include productivity studies.

E. General surveys . .

1. Identify spawning and nursery areas which may come under the influence of plant operation. ,

t

2. Inventory of terrestrial flora and fauna to' identify species and estimated populations. ' ,- .

3. Conduct marshland surveys to establish location of nursery areas and determine the species composition and estimated populations.

This should identify any cyclic or seasonal pattern which r.y be present.

4. Obtain backgrou'nd levels of atmospheric salt content.

Reports and Program Changes [

Quarterly reports will be required. - These reports will be utilized by the staff to judge the adequacy of the progrsm and to determine what changes may be appropriate or necessary to improve the data collection.

These changes will be coordinated with other agencies prior to implementation.

Changes to the. program may be submitted by th'e applicant at any time

- for consideration by the staff. ,

58 .

Y' f

k f

4 e

f I

1 ENCLOSURE B 4

i 1

1 1

i 1

o 4

M

. -c Station locations ( see map)

I. Intake Canal Area A. Area I - 3 stations

1. Stations located in a line perpendicular to the south side of the intake canal and extended approx.

2 miles south of canal.

2. Station location

a. la - in dredge channel immediately south of intake canal.

b. lb .- approx. 3/4 mile further south,

c. 1c - located approx. 2 miles south of intake canal.

B. Area II - 3 stations

1. Stations located in a line perpendicular to intake canal and extending 2 mi. south of canal.

2. Station locations

a. 2a - located in channel near break in northern side of intake canal.

b. Zb - approx. 1 mi. south of intake canal.

c. 2c - approx. 2 mi. south of intake canal.

C. Area III - 3 stations

1. Stations located in a line perpendicular to intake canal and extending 2 miles south of canal.

2. Station locations,

a. '3a - located in channel beyond the physical end of the north side of the intake canal.

1) Placement'of this station based on:

a) practicality of distance.

b) water sampled here should represent the early phase of the entrained water mass.

59 ,

[

- L~s /

1 .  % '#

, ~5'

. s > .el _, c:

3.,,.., .., ,:r:. ......

k

' ..y,

< to

+ ....

"(gm q

)

j22 ee  :::, v .

,,n..v. .

, u y' ; \y%;.. ..... '^'  : ::....

u gv , * ,;, g,,

mul t

%Dn u- .

'k t.. . . . .

,......s

.=n;.:

a::.

R'm q, u / /:" ' I'***'#. ; g , , , . ' , . . , *. , . t ,, ,,, . ' . , . . . J:7.'.);,

,/ e4 W .;;;;.f.

v. ::.. t..

v

...- C'"

iis c s,

(

( s *, g,: l'

'  :..,o....

e s.r., ~ ~ ~ ' .::

"N 1

.:,o:.c?::: . M. . . . . . . :

• tv::*/.*,

.,'.s # ~ ~ ~ .c l*: ,*:. . . . . . ... . . :,*:. :.. . .. .. .. .

'.s ~'~t...,.u~~~~ -

..~

., *l, / %1 . . % *i-

. . &!#.v

==-

- b. O \ **

e '

,;3 ;,, :!*'

N-

3. ::t.. 8:::,.,..s *,. g . *

• s.. . s. .

v) d

. f,.[

e b J

(.S 5

. < m..,

w v :": "::sy:

D .. ...

C'* o

. O < s 2

G z 9 \-

O. .

mammmmmmmmmmm mummm m m mmmm m m mm m m mmmme t

-< Z

.d .,

o -

h e tN t

Q (' 4

. 9

• W .

O*

, .o g < -

• O

. f 60 .

J Samplina Techniaues' I. Area' IV (intake area)

A. Intake pipe

1. .Present 1/2 meter, 2029 mesh nets used to collect zooplankton.

a. Simultaneous replicate samples taken by using bongo nets,

b. Nets towed for 1 minute on surface.

2. MARMAP procedures used to collect fish eggs and larvae.

a. Simultaneous replicate samples taken by using bongo nets.

b. Nets towed for 10 minutes on surface.

B. Mouth of Intake Canal

1. Samples taken at 3 depths.

a.' surface

b. mid-water

c. bottom

2. Sampling procedure same as intake pipe area.

II. Other areas (I, II, III, V, VI)

A. Sampling procedures and net usage same as Area IV.

B. . One depth ( surf ace) sampled.

s .

61 ,

E -

l i

Samoles Collected I. Outs.ide intake and discharge areas (I, II, III, V and VI) =

15 stations x 4 samples / period x 8 periods = 480 samples /

2 wk. period.

II. Intake Area (IV)

A. Intake pipe - 4 samples / period x 8 periods = 32 samples /wk.

B. Mouth of Intake Canal =

4 samples / period x 3 depths x 8 periods = 96 samples /wk.

C. Total samples generated in 2 weeks = 736.

Live-Dead Determinations ( Bi-weekly)

I. Test areas ( discharge side)

A. Samples taken at 2 stations at 3 hr. intervals over 24 hr.

period.

B. 8 samples / period x 8 periods = 64 total samples.

II. Control area (intake pipe)

A. Samples taken at 1 station at 3 hr. intervals over 24 hr.

period.

B. 4 samples / period x 8 periods = 32 total samples.

62

~

1

b. 3b - approx. 1 mile south of intake canal. o l

c. .3c - approx. 2 miles south of intake canal.-

D. Area IV - intake canal ( 2 stations)

1. Stations located at mouth of intake canal ( 4a) and at the intake pipes ( 4b) .

E. Area V - discharge side ( 3 stations)

F. Area VI -- discharge side ( 3 stations)

Sampling frecuency I. Area IV A. Both stations sampled every 3 hourc over a 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period.

B. Samples taken on weekly basis.

II. Areas I, II, III, V and VI A. All stations sampled every 3 hours3.472222e-5 days <br />8.333333e-4 hours <br />4.960317e-6 weeks <br />1.1415e-6 months <br /> over a 24 hour2.777778e-4 days <br />0.00667 hours <br />3.968254e-5 weeks <br />9.132e-6 months <br /> period.

B. Samples taken on biweekly basis.

63

' Field. Collection I.. Area.L IV ~( intake canal) .

A. Need 4 - 3 man ~ shifts; 6 hr./ shift for 24 hrs.

B. ': Total'= 12 men /wk.

.II . - Other . Areas (I, II, III, V, VI)

A..~Need;4-2 man shifts / area; 6 hr./ shift for 24 hrs.

. Total.- 40 men for all areas /2 wk. period.

'B.=

4 I

~

l I

-l

^64:

6 i' . .w+ r e .. , - . . - . , . , , ,, - - - - , , - - - m. -g~.,--- p p--+r-

Manoower Recuirements- Total I.- Field ~ Collections ~

A. Intake area - 12 52 B. Other areas - 40 II. Laboratory personnel ( counting, weighing, sieving) needed in-addition to field crews. 30

._ A . Assuming 1 man / sample /8 hr. day.

1. 1 hr. for sieve separation-

2. 6'hr. for counting organisms

3. 1 hr. for drying and weighing (biomass determination)

- III. Data Processing-A. Computer programmer 1 B. Coders and transcribers 5 IV. - Additional Personnel A. Principal investigator 1 B. Admin. asst. 1 C. Field and laboratory assistants 4 94 f

w -

~

m 65

• ., n- ,

Data Processing I. . Sampling A. . Weekly - 256 B. Biweekly - 480

2. . Biweekly total - 736 D. Yearly total - 19,136 II. Raw Data Sheets A. Number of data sheets / sample = 5 B. Number of data sheets /2 weeks = 3680 C.- Number of data sheets / year = 95,680 III. Coding Sheets A. Calculated on 1 coding sheet / sample B. 25 categories x 4 col./ category = 100 col./ sieve size x 5 sieve sizes = c00 col./ sample plus sequencing and sample information.

IV. Time and monetary requirements to code data A. 10 min./ sample to add columns + 18 min. to code data =

28 min./ sample.

B. 368 hrs, biweekly to code and add = 736 at 2.00/hr.

C.. Yearly dollar total-for coding and adding = 19,136.

D. Manpower needs.for coding and adding = 5 full time employees.

V. Material requirements and costs A. 574 for 19,136 coding sheets (yearly total) .

B. 150.00 to make plate for special coding forms.

C. Total Cost of coding sheets = 724 D.- Computer tapes - 4 at 25 = 100~

E. Total Data set cost including labor and materials - 19,310 66

VI . _ Computer funds 4.: Basic data organization and card punching - 11,500 B. Data' analysis

1. Analysis - biweekly - 150

2. Mapping - biweekly -_150

3. +20% for error and program development

4. Yearly total - 9400 C. Total cost of computer programming - 2 100 VII. Computer programs to be used

.A. SAS - multivariate ANOVA and regression B. S AS -- correlation analysis C. SYMAP - mapping D. .ECOMP - trend surface mapping E .- Prgrams written especially for study VIII. Total Cost'- 40,036 e

67

( a

.. . . - ~ .. - --

/

[

i Estimated Partial Budoet- Total

' I,-II. 82 Marine Biologist-I's at 8200/yr.' 672,400

- III. Data processing. .40,036 i-

. IV. Principal investigator-Admin.-asst. ( Mar. Biol. III) 10,400 Field and Laboratory Asst's. (Mar. Biol. II; at'8600/yr.)' 34,400 V. . Plus equipment (boats, nets, jars, micros-copes, ovens, balances, etc.)  ?

d 4

)

b Au t

t

+

4

'I 68 - l

Day- 1 2 3 4 5 A.M.P.M. A.M.P.M. A.M.P.M. A.M.P.M. A.M.P.M. Sampics generated - 608 26 0 14 40 Sampi s counted - 196

Week 1 52 52 52 52 -52 52 Remainder'- 412 Y Y 13 + 27 + 52. ^ 52 + 52 - 196 Samples generated 128 s 5 Week'2 46 '40 46 52 52 52 52 52 52 52 ' ,,f s 3 ]c d 2 Remainder 292 43 + 49 + 52 ^ 52 + 52 = 243 292 samples /10 days =

26 0 14 40 52 52 52 52 52 52 30 additional peopic Week 3 to process samples V V Y V V 13 + 27 + 52 + 52 + 52 = 196 Week 4 46 40 46 52 52 52 52 52 52 52 V V V V V 43 + 49 + 52 + 52 + 52 - 248 SAMPLING SUDIARY

ENCLOSURE C L.

I. Introduction It has only been in the last ten years or so that the topic

~

of thermal pollution has generated encugh concern to prompt re-scarchers to intensively study its effects on aquatic organisms.

In this time, a vast amount of work has been done on thermal effects, but prienrily concentrating on fishes and commercially important shellfish. Comparatively little work has been done on zooplankton, the pricary consumers that are at the base of most food chains. In a 1968 symposium on thermal pollution, J. B.

Strickland (in Kronkel and Parker 1969) makes a call for research ,

on the after-effects of power plant entrainment on zooplankton.

The few studies that have been done, in fact,seem to have contra-dictory results since they show a spectrum of data reflecting effcces from none at all (thrkowski 1959*) to 80% mortality (Prager 1970, 1971), to even a study shouing 100% mortality (Heinle 1969). Those studies also neglect year-round effects sinco they were done only during the cource of several months in the most critical time or year (summer). A year-round study cf offects

, at any particular site is clearly needed to predict ecological effects upon the area. -

Copepods, being the dominant year round zooplankter by a vast majority, can be studied on a bi-weeh1.y collection basis from June 1973 to June 1974 to predict the effects of entrainment throughout the yearly temperature cycle. Recent techniques have allowed such a study to include data that reflects such things as lethal and delayed lethal effects, offects on reproduction and effects on growth. Subtle population changes caused by any of these offects could have drastic ecological consequences, channel-ing energy from the normal nehton food chains to decomposers, the benthic com= unity or to passive filter-feeders such as cteno-phores (Heilo 1969). Such indirect effects upon the marine commu-nity, in the long run, are as icportant as the direct effects of thermal pollution that have been so extensively studied.

• This study uns done in the colder uaters of England and with questionable techniques.

70

II. Objectives A year long study is proposed that would involve sampling intake and discharge waters on a bi-veehly basis. Sach a study should show year-round effects of power plant entrainment on the major species of copepods, focusing upon (1) percent morta-lity caused by entraincent (2) differential survival between the species and between the age groups of certain species: (3) delay-ed lethal effects due to the period of time spent in the heated effluent of the discharge canal following passage through the plants (4) effects of entrain =ent on reproductions and (5) effects of entrainment on growth of juveniles. Also, data can be obtained to tell about the effects of entrain =cnt other than thermal shock (i.e. turbulence and shearing forces encountered in passing through the condensing tubes) and the effects of tidal recruitment of pai e copopods into the heated waters of the discharge canal without actually going through the plant (Prager 1971).

I 71

1 .

l III. Techniques l l

A.I . To determine the percentage mortality caused by power plant j entrainment and differential survival between the species or age groups of copopods (if any), samples are taken in a metered 50 cm plankton net (80 mesh) from intake and discharge waters. The icngth of the tow depends upon the density of the standing popu .

lation at the particular time of year, but a one-minute tcw will, in general, provide great enough numbers for all purposes dicussed below. One quarter of the intake sample is placed in a gallon container with enough filtered intake water added to make one liter total volume and 1 ml. of'1% neutral red solution is in-jected into this population. The jar is placed in an insulated container of ambient temperature water for 45 min. and the sample is then concentrated in a Clarke-Bumpus net gup, rinsed with tap water and preserved in 10% formalin. One quarter of the discharge

sample is placed in a gallon jar, leveled to one liter with fil-tored discharge water, and also injected with 1 ml. of 1% neutral red solution. This population is incubated 4.n an insulated con-tainer of ambient diceharge unter for 45 min., concentrated, and preserved. This vital staining technique has bean faverably tasted by several laboratories and is discussed by Dressel et a1 (1970). The preserved samples are kept in ice filled insulated containers until they can be counted. The counts are made by first sieve fractionating each sample (the technique used by the current zooplankton study at Crystal River), and then split-ting each fraction enough times so that between 1,000 and 2,000 organisms are counted per sample. The splits are acidified by titrating with a half and half mixcare of SN acedic acid and SN sodium acetate until the neutral red turns red (the formalin and the tap water being slightly basic, cause it to turn yellow).

The split is then placed into two or three gridded petri dishes and counted _under a Wild dissecting scope at 25X power. The cope-pods that were living at the time cf the collection appear dark red (stain uptake occurs in as short of time as five minutes, so this is more or less an instantaneous mortality measurement), while L

i 72

those that were dead appear opaquely white or slightly pink.

The amounts of " dead" and " alive" specimens of each of the major species are tabulated, empicying the use of a manual counted similar to those used for blood counts.

A.2. Percent mortalities can be computed by dividing the number

" dead" counted of any species by the total number of that species.

Total percent mortality for all species is obtained by dividing the number of " dead" specimens counted by the total number of organisms in the split. Using the percent mortalities of the intake population as a control, the discharge percent mortalities above those figures can be attributed to entrainment. A compari-son between the percent mortalities of the various species of copepods should show which are more or less tolerant to the thermal stress.

Since the sieve fractionation separates the age groups of A. tonsa, the percent mortalities for juveniles can be compared with that of adults. Any differential mortality could affect population structure and productivity of the standing crep en the discharge side. If trends in differential mortality are noted, there are methods of analysis that may be employed to determine such effects on community structure (Dr. Frank Nordlie, personal communication).

The total number of organisms in the intake and discharge samples can be estimated by knowing what proportion of the total counted split represented. The volume of water sampled can be calculated by the flow meter readings from the net. Knowing the differences in percent mortaliti.es between intake and discharge, the approricate numbers of the various copepods per cample, the volume of water sampled and the amount of water going through vu.r ime, vae nmn.c en nur or The moc :a-cies r.aua the plant per4hour or per day (under similar co;nditicas)pa could be calculated. This would be useful information if analyzed in conjunction with the zooplankton survey which would estimate the total standing copepod populati.on in the area. Prager, in hi.s 1970 report carri.cd this information one step further by using an estimate of the weight of each copepod to determine the aniount of dead organic materials that were being released in the effluent per day.

73

_i

A.3. As mentioned above, the collectino will be done on a twice a month bacis with all counts being made during the week after the co11cetion (the dye begins to be leeched into the preserving itquid as tire goes on).

l A.4. Raymond Alden, a graduate student, will be involved in co11cetions, counting and sorting of specimens and analysis of the data. Two assistants, who may be undergraduates will assist in collecting (especially boat handling).

A.S. The collecting will involve 8 hrs. every two weeks or so for the graduate student and the assistants. The counting, and data analysis will involve approximately 6 hrs. work for the graduate student between collections. The sieve fractionation3 splitting and care of equipment will require 3-5 hrs for one of the undergraduates. Preparation of fresh stain, acetic acid, l and sodium acetate solutions will require 2-4 additional hours of an assistant's time.

B.I . Dalayed 1cthal effects due to the fact that the copepods are remaining in the heated waters of the discharge canal for hours after entrainment can be tested by the use of a drift- l bottle apparatus (Prager 1971). Another quarter of the dis- l charge sample is placed in a plastic bottle fitted with two 5 4 mesh screens and attached to the apparatus that allows filtered ambient temperature water to be slowly pumped through the sample.

The apparatus is then allowed to follow the currents down the discharge canal for a given period of time (Tarzwell, 1972, suggests a few hours). The sample is then prepared and counted l l in the manner discussed above. In order to have a control for this " delayed" sample, the second quarter of the intake sample l is placed in a flow-through bottle in the intake canal for the I same length of time and then prepared and counted in a similar manner.

1 l

74

B.2. The percent mortality of the " delay" sample and the " delay" control can be computed for the various species as it was for the " intake" and " discharge" samples. These figures can be ana-lyzed in n similar manner as those for the original samples, and probably would represent a truer picture of the situation because the population that does go through the plant doesn't go back into a " normal" environment, but is put under the thermal stress of being in the heated effluent for hours after the original thermal shock. Any increase in cortality between the discharge sample and the " delayed" discharge sample that is not shown as a similar increase in the control canbe interpreted as being due to delayed mortality.

B.3. The collection and counting vill be concurrent with thn e discussed in A.3.

B.4. The collection and counting time, being the same will require only the personnel mentioned above.

B.S. The additional preparation,ccunting and data analysis vill involve 6 more hrs. work per tuo veck period for the graduate student and 3-5 hrs. more work for an assistant.

C.I . J. C. Prager (EPA), at the latest quarterly meeting at Crystal River, suggested that ATP analysis be done as "back-up" data to the above techniques in determining mortality. As this work will be done primarily by Richard Drew, another graduate assistant, he has written up the following section on ATP tech-niques. The ATP data obtained will be analyzed considering the density per liter of animals in intake and discharge samples. A third quarter of each sample vill be used in the ATP sNdies along with splits of the " delay" samples (see B.l. and oelow F.1.).

Proposed _ Technique to Accomnlinh Obiective Use of the ATP assay to determine the viable zooplankton biomass l

75 7

in intake and discharge sampics at the Crystal Riber Power Plant and in samples subject to several hours exposure to discharge

canal temperaturcs.

In studies of conditions of stress on plankton communities,

.it is desirable to measure the viable biomass and at particular times, previous to the stress, during and after o recovery from the stress. In the measurement of zooplankton populations, total organic carbon estimates have been used. However, this method consures all the carbon, which may include detrital ma-terial, and yield erroneous viable biomass measures. DNA has been recently suggested as a possible biomass parameterr Holm-Hansen (1968), used this (DNA) in a study microbial biomass in ocean profiles. He found that DNA was retained to some degree in surrounding particulate matter after cell death, therefore yielding a higher biomass than what existed. Measurements using I respiration or ETS as indicators showed fluctuations which were often unrelated to tha viability of the organism. Adenosine triphosphate has in the last few years become the frontrunner as a measure of biomass viability for several reasons: (1) ATP is found in all living cells: (2) ATP is not found in non-living materials (3) the ratio of ATP to . cell carbon has been found to remain constant over a wide variety of organismsg and (4) there exists a sensitive and rapid assay for ATP. First described by McElroy (1947), the assay is based on a linear relationship between the amount of ATP added to an isolated firefly luciferin-lucifeu a system, and the duration of light, output. In cther words, the amount of luminescence is proportional to the concentration of

. ATP added. In general, the procedure consists of: (1) the ex-  ;

traction of the ATP from the samples (2) the mixing of luciferin-  ;

luciferase enzymes with the ATP samples and (3) measuring the l

! amount of luminescence given off in the reaction.

Field Work l Split sampics will be taken from the' intake, discharge, " delay" l intake. " delay" discharge, and " delay" intake-discharge samples of the given co11cetion period. (The sampling procedures to be f

( ' set up so discharge sampling follows intake sampling by enough L

)

l

! 76

' l

time to allou for sampling the same basic population before and after passage thrcugh the plant.) A sampling period will occur once every tuo weeks for approximately one year, to observe any seasonal variations.

The sattples will be brought back to a trailer laboratory where each split will be filtered through a Mil 11 pore filter (pore-size of 20 micron). The filters are then immediately immerse d in 40.0 ml of boiling Tris /P[ buffer and boiled to a volume of 10 ml. The samples are then frozen (-20'C) and taken back to Gainesville. There each is thawed, mixed, cen-trifuged, and the supernatant poured into a test tube. The luciferin-luciferese enzyme system is added, mixed, and the bio-luminescence is read using a liquid sein tillation spectrometer (Packard Tri Carb Model 2002). Photon emissions are converted to micrograms ATP per liter using previously prepared standard curves. Time required for the entire assay of the 5 sampics is at 1 cast 12-18 manhours /2 weeks.

Analysis of ATP data vs. count densities and percent mor-talities will require approximately 1 hr. Work per week for the graduate student.

D.I . Although laboratory studies on copepod reproduction are admittedly artificial, they may show certain trends of the en-trainment effects in nature. Once every quarter, part of the

" delay" discharge sample, part of the " delay"' intake-discharge sample (see below, E.1.) and part of the " delay" intake sample are brought back to the laboratory alive. Then 100 living adult Acarrio Tonsa (the dominant species in this area) from each sample are placed in 1500 ml covered crystalyzing dishes (20 per dish at constant ambient temperature. The cultures are fed 12h ml.

of a culture containing equal volumes of Chlamydemonas, Rhodomonas baltica, Isochrysis nalbana and Thalassiostra pseudana every two days (Heinle, 1969b).

On days 2, 4, 6, 8, and 10 after the incubation was started, one of the dishes is selected from each of the three sample 77

i cultures, stained, preserved and counted to determine how many adults have survived and how many eggs cr juveniles have been produced. Such a serial observation is necessary to determine uhother the populations are producing less because of decreased survival or decreased fertility and fecundity.

D.2 The amount of juveniles produced per dish can be compared l for the three sampics. The " delay" intake population would be j the control and the other two samples vould represent populations that have undergone thermal stress. Any less production by these populations may be due to adverse affects of this stress.

Of course, any mortality would have to be recorded and the reprou

. duction figured on a per individual basis.

D.3-5~ This task would involve some extra time on collection day every quarter due to the time required to pick out 300 live Acartin tonsa. This time, plus the time required to feed, stain, preservo, and count the sampics would involve 6-10 hrs. for the graduate student and assistant. Also, cultures of the algae must be made and maintained, taking approximately 4-6 brs. more time per month to weigh and autoclave chemicals for culture media.

E.1. To determine the effects of entrainment on the growth of juveniles, a technique modified from that used by Heinle (1969) is employed. Water from the intake and discharge canals are poured through #73 Nitex bolting cloth (.073 mm aperture) into plastic gallon jars fitted with 5 micron mesh screened windows.

In this way, only eggs and first naupliar stages of Acartia tonsa pass through the cloth and remaintrapped in the jars. The amount of water poured through this system depends on the density at the particular time of year (at this time of year 12 gals. will provide approxicately 200 juveniles in this manner). Since water going through the plant is a mixture of water near the surface and water near the bottom, the sampling from the intake side must be made in such a way as to adjust for this fact. The water sampled 78 1  !

can be taken by a Nansen bottic from water near the bottom in several foot intervals, to water near the surface. The intake and discharge sampics are brought back to the laboratory in in-sulated containers of. ambient temperature water and each split (using a Folsom Palnkton Splitter) into 8 portions. Each split is placed in a jar, leveled to a liter with filtered intake vator, dnd incubated in an illuminated B.O.D. box at constant

. ambient (intake) temperature 4 These cultures are fed 10 ml.

of the algal cultures centioned above every tuo days. On days 0, 1, 2, 3, 4, 5, 6, and 14 after incubation, jars of the intake and discharge samples are picked at random to be strained, pre-served and counted.

E.2. The mean stage attained by the juveniles by each day can be compared between the intake and discharge populations. Growth curves can be made to compare the tuo populations and second ge-neration production may be obtained Ln comparing the day 14 sam-ples. If there is large enough numbers to permit, dryteights could be measured as a parameter of size attained by the adult, A. tonsa and a comparison of mean weights between the populations would show the ultimate offect on growth.

E.3. The samples are taken at the end of each co11ceting day mentioned above. The second assistant will be making this collec-tion while the graduate student and the assistant are making collections described in "A" and "B" and following the drift bottle apparatus.

E.4,5 The graduate student and assistant are required to split the sampics, feed the splits every two days, stain, concentrate, preserve and count (includes identification of stages) the 8 splits of each sample. This time, plus data analysis and dry weight measurements may,take 20-25 h~ours betwden every. collection ported.

79

?

l i

F.1. To test the effects of entrainment other than thermal stress and to tell comething about the effects of tidal recruitment,

( Prager (1971) has suggested that part of the intake sample (the remaining quarter) be placed in another bottle on the drift-bottle apparatus. This sample will be exposed to essentially l

the same thermal conditions as the " delay" discharge cample popu-lation without the turbulence of power plant passage and could represent a population that has been pulled into the thermal of-i fluent by tidal currents. This population is also allowed to remain in the drift bottle for several hours, stained, preserved, and counted.

r F.2. A comparison can be made with the " delay" discharge sample percent mortality to see how much of the mortality is due to thermal shock and how much is due to mechanical damage (although, admittedly, thermal stress may lower an organism's tolerance against mechanical damage). It also tells something about how dangerous the heatcd water in the discharge canal is to the cepe-pod popu3ntion in the area during incoming tides. - . -

F.3. This collection will also be done on collection days.'

F.4-5. This additional preparation count and analysis will add 3-4 hrs. Work for the graduate student and assistant.

G.I. During the summer quarter, when more time is available and when the thermal situation is more critical, several addi-tional tasks wi11 be attempted.

First, at least one 24-hr long sampling period will be used to take samples of eggs (see above) at three hour intervals from the intake and discharge canals (Heinic, 1969). These will be incubated, prepared, and counted (as above) to get more complete data on the effects of the en-trainment throughout the day and night of the most critical time of year. -

80

)

1 G.2. A seties of grouth curves can be made to compare intake and discharge populations and daytime and nightime samples.

G.3-5. This 24 hr. period would be mid-summer and would in-volve the graduate student and the assistant (s). The weeks follouing the collection vould involve culturing, preparation and counting of 96 sampics which would involve between 115-120 hrc. work for the graduate student and assistants. Data analysis may involve 10-15 hrs.

i H .1. Another task that may be carried out during the summer is laboratory simulation studies upon thermal shock of the type found in passage through the power plant at the various times of year. These studies can be done by placing 5-10 copepods (accilimated to any given " intake" temperature) in 50 ml. test tubes. filled with filtered intake water. Half of such tubes would be submerged in a hot water bath and raised to discharge temperature for the particular time of year (winter, spring, summer) to be simulated. The other half of the tubes would be placed in ambient temperature sea water. Both sets of tubes would reamin at the given " intake" and " discharge" temperatures j for 2 hours2.314815e-5 days <br />5.555556e-4 hours <br />3.306878e-6 weeks <br />7.61e-7 months <br /> and the copepods then would be incubated at ambient temperature in crystalyzing dishes and fed as above. Upon staining, preserving and counting after 7 days, mortality would be compared, as well as any reproduction. Such tests could be repeated with various stages of A. tonsa obtained in a manner like that discussed in the growth studies, thus allowing further study of the effect of the thermal shock upon growth.

H.2. Analysis of this data would be similar to those of the field studies, only much closer observations can be made as far as effects are concerned because the types of organisms can be chosen and conditions made identical except for the variable of temperature.

1

+

81

~

_ . .L . _

H.3-5. This additional study must be kept flexible as far as )

amount of tie.c required because only one graduate student is available for counts and thi.s part of the study must be made to fit around the field study.

I.1. If possible, an additional collecting day per month will be added during the critical summer months. This would, of course, increase manhours required accordingly. Overlap time on culturing equipment and variable time required for other summer studies necessitate that these extra collecting days be optional.

Extra manhours vill be used in such additional studies whenever possibic.

J. Studies involving sorting of live and dead organisms from splits,of Intake and Discharge samples by direct observation at the plant site may be done from time to time as a second check on the craining technique. An assistant would perforia the sort--

ing (as doccribcd by Prager 1970) and preserving, while the gra-duate student would do counts and identification of the " alive" and " dead" portions back in the lab. This would required variable amounts of time, depending on how many times such " checks" were done.

I 1

82 1 1

IV. Capital Equipment and Expenses A. The metered 50cm plankton net (80 micron mesh) is used for intake and discharge sampling. Price: $141.80 B. A Folsom Plankton Splitter for spittting live juveniles into equal portions for grouth study and fcr splitting intake and discharge populations into splits to be stained, those-to be used in ATP studies and those to be used in the drift-bottle apparatus. A splitter is already available for splitting pre-served materials. Price: $186.00 C. Clarke-Dumpus type net cup serves dual pur;ose as cod end container of plankton net (to avoid mechanical damage to copepods upon capture -- see Prager,1971) and to concentrate sampics prior to preserving. Price: $26.30.

D. Dissection scope for counts.

E. B.O.D. box with-light source for culturing juvenile and adult A. tonen at constant ambient temperature throughout the year. Prices $995.00 F. Counter G. Nansen bottle H. Hot water bath- can be made rather inexpensively by using an immersion heater with a-thermostat in a tank or using a Temp-Blok module heater. Prices $55.00 - $60.00 l

I. Transportation to and from Crystal River for the 3 or 4 work-ers and their equipment (leased fram 0 of F).

J. A boat will be required during the bi-weekly collections l l

l l

K. On site laboratory space will be required for the ATP analysis and some of the work with sorting of dead and live organisms.

83

L. Since Richard Drew is doing ATP analysis on equipment obtained by a Florida Pouer Corporation grant, the only equipment needed for this will be chemicals for one years' worth of tests. Approx.

Cost: $50,00 pluse cost of Liquid Nitrogen coolant.

M. Misec11ancous 14 96 L. jars - approx. $60.00 24 1 doz. culture dishes - approx. $35.00 34 5 micron filter bags - approx. $20,00 - -

4. Carboy and plastic gallon containers - approx. $25.00

5. #73 bolting cloth - $20.00-

6. Gridded petri dishes - $27.00 74 50 ml. test tubes - $45.00 84 pipettes - $20,00 .

9. Acetic acid, sodium acetate, neutral red, and formalin

- $20.00

10. Chemicals for algae culture media - $30.00 124 Insulated container (ice coolers) - $20,00

13. Micro-di.'section Kit - $20.00

14. Materials to build drift bottle - $100.00 (if disposable pumps are used)

15. Per diem for 3-4 persons' for 24 hr. collecting trip

- $18.00 N. Salarios

1. Raymond Alden, graduate student - 9 months time research assistantship - $4,005.00 and 3 months (summer) full-time as Marine Biologist II - $1,800.00. Total 12 months

$5,805.00.

2. Richard Drou, graduate student. 9 months 1/3 time re-search assistantship - $3,150.00 and 3 months (summer) time research assitantship - $1,224.00 - Total $4,374.00.

3. Two half-time undergraduate assistants working up to 15 hrs /wk. each at $200/hr. up to $3,120.00

4. Total salaries - $13,299.00 84

V. Budget ,

1, Salaries $13,300.00 ,

2, Equipment $1,960.00 (approx.)

3. Total budget $15,260.00 e

85

_